PMID- 12848652 TI - Benign pulmonary metastasizing leiomyomatosis in pregnancy: a rare complication after cesarean section. PMID- 12848653 TI - Prenatal diagnosis of fetal hydrops associated with Down's syndrome in a 40-year old woman with a mosaic Turner's karyotype (45,X/47,XXX). PMID- 12848654 TI - Spontaneous pregnancy in a 45,X/46,X,r (X) Turner's mosaic patient. PMID- 12848655 TI - Celiac disease in pregnancy--not always a relapse. PMID- 12848657 TI - The incidence of diastolic right atrial collapse in patients with pleural effusion in the absence of pericardial effusion. AB - The aim of this study was to describe the incidence of cardiac chamber collapse assessed by echocardiography and explore possible mechanisms in a clinical population of 116 patients with pleural effusion in the absence of pericardial effusion. We found that the frequency of chamber collapse was 18% in patients with pleural effusion in the absence of pericardial effusion, thus cardiac chamber collapse occurs in patients with pleural effusion. PMID- 12848658 TI - Echocardiographic features of patients with heart failure who may benefit from biventricular pacing. AB - BACKGROUND: Recent studies suggest that cardiac resynchronization therapy through biventricular pacing (BVP) may be a promising new treatment for patients with advanced congestive heart failure (CHF). This method involves implantation of pacer leads into the right atrium (RA), right ventricle (RV), and coronary sinus (CS) in patients with ventricular dyssynchrony as evidenced by a bundle branch block pattern on electrocardiogram (ECG). Clinical trials are enrolling stable patients with ejection fractions (EF) /= 54 mm, and QRS duration >/=140 msec. We compared echocardiography features of these patients (group 1) with other patients with EF /= 54 mm, and QRS < 140 msec (group 2 = presumably no dyssynchrony). METHODS: Nine hundred fifty-one patients with CHF, LVID 54 mm, EF 35% by echocardiography were retrospectively evaluated. One hundred forty-five patients remained after those with primary valvular disease, prior pacing systems, or chronic atrial arrhythmias were excluded. From this group of 145 patients, a subset of 50 randomly selected patients were further studied (25 patients [7 females, 18 males] from group 1, and 25 patients [7 females, 18 males] from group 2). Mean age group 1 = 75 years old, mean age group 2 = 67 years old. Mean QRS group 1 = 161 msec, mean QRS group 2 = 110 msec. Each group was compared for presence of paradoxical septal motion, atrial and ventricular chamber sizes, LV mass, LVEF, and RV systolic function. RESULTS: Of the initial group of 951 patients, 145 (15%) met inclusion criteria. In the substudy, 20/25 (80%) of group l and 7/25 (28%) of group 2 subjects had paradoxical septal motion on echo (Fisher's exact test, P = 0.0005). The t-tests performed on the other echocardiography variables demonstrated no differences in chamber size, function, or LV mass. CONCLUSIONS: Cardiac resynchronization therapy with BVP appears to target a relatively small population of our advanced CHF patients (15% or less). Although increasing QRS duration on ECG is associated with more frequent paradoxical septal motion on echo, it is not entirely predictive. Paradoxical septal motion on echo may therefore be more sensitive at identifying patients who respond to BVP. Further prospective studies are needed. PMID- 12848659 TI - Ejection fraction: subjective visual echocardiographic estimation versus radionuclide angiography. AB - OBJECTIVES: The aim of this study was to validate the correlation of left ventricular ejection fraction (LVEF) between two-dimensional (2D) echocardiography with harmonic imaging modality and radionuclide angiography (RNA) in a large number of patients. BACKGROUND: Subjective visual estimation of LVEF is the most prevalent method in evaluating left ventricular systolic function. Previous published reports demonstrated that visual estimation from 2D was superior to other mathematical calculations, however the studies contained less than 50 patients. METHODS: Retrospective review of 377 consecutive medical records containing LVEF from both 2D echocardiography and RNA obtained from the same patient within 15 days from January 1998 to September 1999. RESULTS: Correlation coefficient value (r) between 2D echocardiography subjective visual estimation and RNA was 0.84. The r value was further increased to 0.87 when heart transplant recipients excluded. The agreement of LVEF between 2D echocardiography and RNA are higher in normal or severely depressed LV. CONCLUSION: Excellent correlation of LVEF exists between 2D echocardiography subjective visual estimation and RNA, especially in normal or severely depressed LV. PMID- 12848660 TI - Estimation of left ventricular ejection fraction from Doppler derived myocardial performance index in patients with acute myocardial infarction: agreement with echocardiographic and radionuclide measurements. AB - OBJECTIVE: The aim of the study was to validate a previously published method to calculate left ventricular ejection fraction (EF) from the myocardial performance index (MPI or Tei-index) in patients with acute myocardial infarction (MI). METHODS: Sixty-one patients in sinus rhythm without overt heart failure were examined between 2 and 7 days after the acute MI. Doppler tracings from mitral inflow and left ventricular outflow were recorded together with two-dimensional echocardiographic (2DE) recordings. MPI was calculated from the Doppler tracings, and EF measured with the biplane Simpson's method. From MPI the EF was calculated by the formula EF = 0.60 - (0.34 x MPI). Radionuclide angiographic (RNA) measurements of EF were performed within 1 day of the Doppler echocardiography. RESULTS: Compared with radionuclide EF, MPI derived EF significantly underestimated EF by 0.03 (+/-0.013; P = 0.027), whereas there was no significant difference in mean EF between 2DE and RNA. There was no statistically significant difference in the agreement between MPI derived EF relative to RNA, or 2DE relative to RNA. The agreement between the three methods was only moderate with wide limits of agreement (+/-0.17). The relationship expressed by the proposed formula for calculating EF from MPI was not statistically significant in regression analysis in this patient population. CONCLUSIONS: No statistically significant relationship was found between MPI and EF by radionuclide angiography. However, MPI derived EF was as accurate as biplane echocardiographic measurements of EF when compared with radionuclide EF, but the agreement between methods was only moderate. PMID- 12848661 TI - Relationship between noninvasive reperfusion criteria and pulsed-wave tissue Doppler parameters in patients with acute myocardial infarction receiving thrombolytic therapy. AB - Several studies have proven that noninvasive reperfusion criteria (NRC) have prognostic significance in patients receiving thrombolytic therapy (TT) after acute myocardial infarction (acute MI). In this study, we investigated the relationship between NRC and pulsed tissue Doppler (PTD) parameters in patients receiving TT after acute MI, and the role of PTD in the management and follow-up of patients with acute MI. The study group(n= 41)was divided into four subgroups defined as: anterior and posterior MI, with or without NRC. In the first PTD measurements (2-3 days after acute MI), all acute MI patients had significantly smaller peak systolic (S-wave) velocity in all evaluated segments and longer Q Speak durations (time elapsed from the inscription of the Q-wave on the surface ECG to the peak of the S-wave in PTD) as compared with control patients(n= 22; P < 0.001 for both). Among the diastolic parameters, the E/A ratio was significantly smaller in the study group compared with the control group(P < 0.001). Among the patients who had received TT in the first 2 hours, those patients who had NRC displayed significantly higher peak S-wave values in all evaluated segments than those without NRC(P < 0.05). The second PTD study (4-5 weeks after acute MI), revealed that the difference between the systolic PTD parameters of the noninfarcted regions of the study and control groups disappeared. Infarct-related segments, however, displayed significant improvement only in patients having NRC. There was a significant positive correlation between the mean mitral annular S-wave velocity and left ventricular ejection fraction(r = 0.59, P < 0.001). In conclusion, a significant relationship was observed between the PTD parameters and the NRC, which are known to have prognostic significance. PMID- 12848662 TI - Does the myocardial performance index affect pulmonary artery pressure in patients with mitral stenosis? A tissue Doppler imaging study. AB - BACKGROUND: The relation between systolic pulmonary artery pressure (PAP) and mitral stenosis (MS) has been poorly understood. Although the mitral valve area (MVA) is an important factor affecting the PAP, there is a wide spectrum of the PAP in patients with MS despite a similar MVA. So, we analyzed whether the left and right ventricular myocardial performance index (MPI) correlated with the PAP. METHODS: Two-dimensional Doppler echocardiography was performed in 46 patients with MS. The left atrial diameter, mean mitral gradient, and MVA were measured. The PAP was derived from the tricuspid regurgitant jet velocity. The ejection time (ET), isovolumetric relaxation time (IRT), and contraction time (ICT) were measured on annulus of interventricular septum, lateral, inferior and anterior wall of left ventricle, and right ventricle free wall from apical two- and four chamber views in patients with MS and 40 age-matched healthy patients by tissue Doppler imaging (TDI). Then the MPI was calculated as (IRT + ICT)/ET for both left and right ventricle. The correlation of PAP with MVA, mean mitral gradient, left atrial diameter, and left and right ventricular MPI was evaluated. RESULTS: MVA and PAP were measured as 1.57 +/- 0.39 cm2 (0.8-2.5 cm2)and 42 +/- 16 mmHg, respectively. It was determined that the MPI increased in patients with MS(0.59 +/- 0.1 vs 0.48 +/- 0.07, P < 0.001). It was also demonstrated that the MVA, left atrial diameter, mean diastolic gradient, and left ventricular MPI were correlated with PAP(r =-0.39 [P = 0.007], r = 0.43 [P = 0.003], r = 0.58 [P < 0.001], and r = 0.65 [P < 0.001], respectively). In multivariate analysis, although the PAP correlated with mean diastolic gradient and MPI (r = 0.39 [P = 0.013], and r = 0.48 [P < 0.001]), it did not correlate with left atrial diameter and MVA. The PAP also correlated with right ventricular MPI(r = 0.63 [P < 0.001]). CONCLUSION: This study demonstrates that the left ventricular MPI obtained by TDI is an important marker of PAP, and right ventricular MPI correlates with the PAP in patients with MS. PMID- 12848663 TI - Lower receiving frequencies than transmitting frequencies could yield improved results for contrast imaging: an in vivo study in closed chest canines. AB - BACKGROUND: Ultrasonic imaging methods of receiving at higher frequencies, which are multiples of the transmitting frequencies (harmonic imaging), are well established as a means of improving myocardial visualization in association with intravenous contrast administration. This exploratory study examined the effect of using receive frequencies that were lower than the transmit frequencies while imaging closed chest dogs with an Ensemble wideband, phase inversion contrast program on a modified Siemens Elegra scanner. METHODS: Intravenous bolus injections of 0.75 mL Definity and 1 mL QW7437 were administered to six anesthetized dogs. Intermittent imaging for contrast visualization was performed using either a broadband array, transmitting at 1.4 MHz and receiving at 2.6-3.2 MHz or a broadband 4-7.5 MHz transducer transmitting at 6.0 MHz and receiving at 4.2-4.5 MHz. Contrast enhancement was measured by videodensitometry, sampling mid cavity and within the myocardium before and after injection. The changes in videodensity from control to after injection were calculated for each method. RESULTS: There was no significant difference in the change in intracavity videodensity between the two imaging strategies although there was near full intracavity saturation in all cases. However, the change in myocardial density was significantly greater for both contrast agents when using receiving frequencies lower than transmitting frequencies (P = 0.02 and 0.03). The difference in duration of the myocardial blush did not reach statistical significance but it tended to persist for longer with the lower receiving frequencies. CONCLUSION: Delivering sound energy at a slightly higher frequency and receiving at lower than the transmit frequency may be an advantageous method of enhancing myocardial perfusion signals during intravenous contrast echocardiography. PMID- 12848664 TI - Dynamic three-dimensional color flow Doppler: an improved technique for the assessment of mitral regurgitation. AB - BACKGROUND: Prior studies have reconstructed mitral regurgitant flow in three dimensions displaying gray scale renditions of the jets, which were difficult to differentiate from surrounding cardiac structures. Recently, a color-coded display of three-dimensional (3D) regurgitant flow has been developed. However, this display was unable to integrate cardiac anatomy, thereby losing spatial information, which made it difficult to determine the jet origin and its spatial trajectory. To overcome this limitation, an improved method of 3D color reconstruction of regurgitant jets obtained from color flow Doppler using a transesophageal approach was developed to allow the combined display of both color flow and gray scale information. OBJECTIVES: To demonstrate the feasibility of 3D reconstruction of regurgitant mitral flow jets using an improved method of color encoding digital data acquired by transesophageal echocardiography (TEE). METHODS: We studied 46 patients undergoing a clinically indicated TEE study. All subjects had mitral regurgitation detected on a previous transthoracic study. Atrial fibrillation or poor image quality were not used as exclusion criteria. The 3D study was performed using a commercial ultrasound imaging system with a TEE probe (Sonos 5500, Agilent Technologies). A rotational mode of acquisition was used to collect two-dimensional (2D) color flow images at 3-degree intervals over 180 degrees. Images were processed off line using the Echo-View Software (TomTec Imaging Systems). Volume-rendered 3D color flow jets were displayed along with gray scale information of the adjacent cardiac structures. RESULTS: Mitral regurgitant flow, displayed in left atrial and two longitudinal orientations, was successfully reconstructed in all patients. The time for acquisition, post processing, and rendering ranged between 10 and 15 minutes. There were 28 centrally directed jets and 15 eccentric lesions. Eight patients in the study had periprosthetic mitral regurgitant flow. CONCLUSIONS: Three-dimensional imaging of mitral regurgitant jets is feasible in the majority of patients. This improved technique provides additional information to that obtained from the 2D examination. Particularly, in patients with paravalvular leaks 3D color flow Doppler provides information on the origin and the extent of the dehiscence, as well as insight into the jet direction. In addition, in patients with eccentric mitral regurgitation, this new modality overcomes the inherent limitations of 2D echo Doppler by depicting the full extent of the jet trajectory. PMID- 12848665 TI - Ankylosing spondylitis in association with mitral stenosis, mitral regurgitation, and aortic regurgitation: a case report and review of the literature. AB - We report on a patient with ankylosing spondylitis in association with mitral stenosis, mitral regurgitation, and aortic regurgitation. Despite extensive search of literature, we could not find association of mitral stenosis with ankylosing spondylitis. This report is the first to describe this association. Our findings are based on clinical and echocardiographic findings. PMID- 12848666 TI - Diastolic paradoxic jet flow in patients with hypertrophic cardiomyopathy: report of two patients with different morphologic aspects. AB - BACKGROUND: In non-Asiatic population, apical hypertrophic cardiomyopathy represents 1% to 2% of hypertrophic cardiomyopathies. The presence of an apical ventricular aneurysm occurs in 1% of patients with the apical variant of hypertrophic cardiomyopathy. METHODS AND RESULTS: We present two patients with hypertrophic cardiomyopathy, both illustrating an early diastolic paradoxic jet flow between the apex and the middle area of ventricle. Such paradoxic flow is possibly due to the coexistence of mid-chamber obliteration and segmental wall motion abnormalities at the apex. The incidence and clinical significance of this flow pattern is still to be clarified. PMID- 12848667 TI - Successful repair of aortic and mitral incompetence induced by methylsergide maleate: confirmation by intraoperative transesophageal echocardiography. AB - Methylsergide maleate, an effective anti-migraine medication, has a well documented association with left-sided cardiac valve dysfunction. Prior reports have described cardiac valve dysfunction in patients using methylsergide chronically for a minimum of 6 years, with surgical intervention consisting of valve replacement for patients with intractable congestive heart failure. We report a 51-year-old woman who developed severe mitral and aortic valvular dysfunction after taking methylsergide maleate for migraine headaches for a period of 19 months, and who subsequently underwent aortic and mitral valve repair with excellent short-term results. PMID- 12848668 TI - Role of transesophageal echocardiography in detecting implantable cardioverter defibrillator lead infection. AB - Implantable cardioverter defibrillator (ICD) lead infection is a rare condition with reported incidence of 0.2% to 16%. It usually presents with persistent bacteremia or fever of unknown origin and requires high clinical suspicion for diagnosis. Whenever ICD lead infection is suspected, transesophageal echocardiography is the diagnostic technique of choice for detection and characterization of the lesions. Lead infections are extremely difficult to manage conservatively and surgical removal of the entire defibrillator system is recommended along with antimicrobial therapy. We describe a case of recurrent staphylococci bacteremia due to an ICD lead infection in a patient with arrhythmogenic right ventricular dysplasia. PMID- 12848669 TI - Alteration of surgical management following intraoperative transesophageal echocardiography in a patient with mobile left atrial thrombi embolized during anesthesia. AB - This report describes alteration of surgical management in a patient with a large free-floating ball thrombus in the left atrium, which was embolized before sternotomy as demonstrated by intraoperative transesophageal echocardiography. Due to new findings observed by intraoperative transesophageal echocardiography, unnecessary sternotomy could be avoided. PMID- 12848670 TI - Hoarseness, hemoptysis and a hole in the aorta: a case review. PMID- 12848671 TI - Pacemaker endocarditis with sizeable vegetation and intermittent fever. PMID- 12848672 TI - Transcatheter closure of an atrial septal defect within a giant aneurysm of the fossa ovalis. PMID- 12848673 TI - Platypnea-orthodeoxia in a patient with ostium primum atrial septal defect with normal right heart pressures. AB - We describe an adult patient with an ostium primum atrial septal defect (ASD) and a patent foramen ovale (PFO) with normal right heart pressures who presented with platypnea and orthodeoxia. A dilated aortic root encroaching into the region of the ASD and PFO, along with a tricuspid regurgitant jet moving into the left atrium through the ASD found on the transesophageal echocardiogram may have been responsible for orthodeoxia. Surgical closure of these defects resulted in the disappearance of both platypnea and orthodeoxia. PMID- 12848674 TI - Transesophageal three-dimensional echocardiographic demonstration of Ebstein's anomaly. AB - We report three-dimensional transesophageal echocardiographic findings in an adult patient with Ebstein's anomaly. Using the anyplane technique and multiple views, especially the short-axis view of tricuspid valve, three-dimensional transesophageal echocardiography clearly demonstrated the intermittent tethering of all three leaflets of tricuspid valve to the right ventricular walls giving a "bubble-like" appearance. On the other hand, two-dimensional transesophageal echocardiography demonstrated well the tethering of the septal tricuspid leaflet, but tethering of the other two leaflets was not well seen. To our knowledge, these findings have not been demonstrated by three-dimensional transesophageal echocardiography before. PMID- 12848675 TI - Echocardiographic left ventricular mass in African-Americans: the Jackson cohort of the Atherosclerosis Risk in Communities Study. AB - Characterization of target organ damage from hypertension is of particular interest in African-Americans, and evidence from electrocardiographic studies suggests that left ventricular hypertrophy is a frequent clinical finding of considerable prognostic importance. Echocardiographic studies may permit more precise characterization of the pathologic impact of hypertension on cardiac structure and function. The objective of this study is to characterize left ventricular (LV) structure including measures of wall thickness, septal thickness, internal dimension, and mass in a middle-aged sample of African Americans using echocardiography. This study is a cohort (cross-sectional) study in which 2445 middle-aged African-American study participants from a population based sample initially enrolled by the Atherosclerosis Risk in Communities, Jackson, Mississippi Examination Center in 1987-1989 underwent an M-mode echocardiograpic examination at their third or fourth clinic visit in 1993-1996. Measures of LV mass, even where indexed by size were conspicuously greater in men compared to women, and men exhibited a demonstrably steeper gradient of LV mass across the rather restricted age range of the study. However, when gender specific thresholds for LV hypertrophy were utilized, African-American men appear to have lower prevalence of LV hypertrophy than women. The lowest prevalence of LV hypertrophy was observed in African-American men who did not have hypertension (28.4%). The findings confirm previous suggestions from electrocardiographic investigations that cardiac hypertrophy is common, if not epidemic in middle-aged African-American men and women, whether or not they have hypertension. PMID- 12848676 TI - Course of intraatrial thrombi resolution using transesophageal echocardiography. AB - Thromboembolic events are associated with atrial fibrillation and with cardioversion to sinus rhythm. Although studies have demonstrated the risk of this complication is reduced by a 3-week period of anticoagulation prior to cardioversion, limited data have suggested a longer period of anticoagulation is necessary for thrombus resolution. We identified and followed 25 patients noted to have intraatrial thrombi on an initial transesophageal echocardiogram (TEE) who subsequently had a follow-up TEE. The majority of patients had a single thrombus, often but not uniformly located in the left atrial appendage with the largest found in those patients with mitral stenosis. Repeat TEE was performed at a mean of 4 +/- 6 months and persistent thrombus was noted in 19 of 25 patients (76%). Seven of 19 patients with persistent thrombi were cardioverted and one of these patients had a neurologic event following the procedure (14%). The only findings associated with persistent thrombus were the presence of mitral valve disease and atrial fibrillation. Our findings suggest that intraatrial thrombi do not generally resolve following several weeks of anticoagulation and that persistent left-sided intraatrial thrombi may be associated with an increased risk for events following cardioversion. Given that a TEE-guided approach to cardioversion is being utilized more frequently, it may be important to determine thrombus characteristics on follow-up that would be predictive of embolic events following cardioversion. PMID- 12848677 TI - Pulmonary venous flow in pure mitral stenosis and sinus rhythm--does pulmonary hypertension alter pulmonary venous flow velocity? AB - Pulmonary venous flow (PVF) is influenced by changes in left atrial (LA) pressure and function in various diseases. In mitral stenosis (MS), there is an alteration of LA hemodynamic due to the impaired ventricular filling caused by the stenotic valve and elevation of LA pressure. Although a variety of altered patterns of PVF have been described in MS, the potential influence of pulmonary hypertension, which is frequently associated with MS and has an adverse effect on the functional status and the prognosis of MS, on the PVF pattern is not clearly defined. The aim of this study is to determine the effects of pulmonary hypertension on PVF in patients with MS. Thirty-eight consecutive patients with pure MS and sinus rhythm (30 females, mean age 40 years old) underwent transthoracic and transesophageal echocardiography. Right heart and transseptal catheterization was also performed to measure pulmonary artery systolic pressure (PASP), pulmonary artery diastolic pressure (PADP), pulmonary capillary wedge pressure (PCWP), and left atrial pressure (LAP). The subjects were divided into two groups: group 1 (n = 25)included subjects with PASP < 50 mmHg, group 2(n = 13)included subjects with PASP 50 mmHg. LA size, mitral valve area (MVA), mean mitral gradient (MG), LAP, PASP, PADP, PCWP, and the peak velocity of PV systolic (PVFS), diastolic (PVFD) and atrial reversal flow (PVFAr) were also measured and compared between the two groups. There was no significant difference in age, heart rate, MVA, and LA size between the two groups. LAP, PASP, PADP, PCWP, and MG were significantly higher in group 2. However, PVFS and PVFAr were significantly lower in group2 (46.6 +/- 15.8 vs 29.9 +/- 12.8 cm/sec, P < 0.005; 22.1 +/- 6.8 vs 17.3 +/- 5.9 cm/sec, P < 0.05). There was no significant difference of PVFD between the two groups. PVFS had negative correlation to LAP, MG, PASP, PADP, and PCWP. PVFAr had negative correlation with PASP, PADP, and PCWP. There was no significant correlation between PVFAr and MVA. LA size and LAP. In conclusion, in patients with pure MS and sinus rhythm, PVF, especially PVFAr, is influenced by the presence of pulmonary hypertension. PMID- 12848678 TI - Quantitation of papillary muscle function with tissue and strain Doppler echocardiography measures papillary muscle contractile functions. AB - It is well known that papillary muscle dysfunction may cause mitral regurgitation, however, there is not a standard technique to measure the papillary muscle function. The aim of this study is to investigate papillary muscle function by using tissue and strain rate Doppler echocardiography. METHOD: A study group (SG) of 42 patients (12 females, 30 males, mean age: 53.8 +/- 10.9 years) with coronary artery disease who have segmental left ventricular dysfunction and mitral regurgitation, and as a control group (CG) 20 patients (6 females, 14 males, mean age 52.5 +/- 10.2 years) who have no coronary artery disease and mitral regurgitation were studied. A subgroup 20 patients (7 females, 13 males, mean age 42.5 +/- 7.5 years) who have rheumatic mitral regurgitation was included to the study. The patients who have left ventricular and mitral annular dilatation were not included into the study. Longitudinal wall motion of mid-inferior segment (one point) and posterior papillary muscle (two point) of the left ventricle was assessed by tissue and strain rate Doppler echocardiography from the apical two-chamber view. Systolic tissue velocity (Sm), strain rate (SR), and strain (S) were measured from the two points in all patients. RESULTS: Comparing the one point values Sm, SR, and S were found less in SG than CG (respectively 4.2 +/- 1.4 vs 11.3 +/- 1.9 cm/sec, -1.0 +/- 0.4 vs 1.7 +/- 0.3 s-1, -13.5 +/- 5.0 vs -18.1 +/- 3.9%; P < 0.001 for all). Comparing the two point values Sm, SR and S were found less in SG than CG (respectively 2.8 +/- 1.2 vs 3.7 +/- 2.0 cm/sec, -0.8 +/- 0.4 vs -1.6 +/- 0.3 s-1, -9.7 +/- 4.6 vs 15.8 +/- 3.7%; P < 0.001 for all). Systolic mitral annulus, left ventricular systolic and diastolic dimensions were similar in two groups. Comparing the CG and subgroup, there were no significant differences with respect to Sm, SR, and S measured from the two points. CONCLUSIONS: Papillary muscle function may be assessed quantitatively by using tissue and strain rate Doppler echocardiography. Tissue velocity, strain rate, and strain values were found decreased in dysfunctional papillary muscle. PMID- 12848679 TI - Quantitative assessment of short axis wall motion using myocardial strain rate imaging. AB - Although left ventricular wall motion has been usually assessed with four-point scale (1 = normal; 2 = hypokinesis; 3 = akinesis; 4 = dyskinesis) based on the visual assessment, this method is only qualitative and subjective. Recently, a new echocardiographic system that enables calculation of myocardial strain rate based on tissue Doppler information has been developed. We investigated whether myocardial strain rate could quantify regional myocardial contraction in 17 patients with and without wall motion abnormalities including 6 patients undergoing dobutamine stress echocardiography. Left ventricular short-axis wall motion was assessed with standard two-dimensional echocardiography at basal, mid ventricular, and apical levels. The same levels were imaged with tissue Doppler method to determine regional myocardial strain rate. Sixty-four segments were judged normokinesis, 53 segments hypokinesis, and 18 segments akinesis at rest; 16 segments were judged normokinesis and 6 segments hypokinesis at stress. No segments characterized dyskinesis. Strain rates of normokinetic, hypokinetic, and akinetic wall segments at rest were significantly different each other (-2.0 +/- 0.6 for normokinesis,-0.6 +/- 0.5 for hypokinesis,P < 0.0001 vs. normokinesis, and-0.008 +/- 0.3 for akinesis, P < 0.0001 vs. normokinesis and hypokinesis). Further, strain rates well reflected the change in wall motion induced by dobutamine challenge: strain rates in the 15 segments revealing augmented wall motion changed from -2.0 +/- 0.7 to -4.7 +/- 1.7 (1/sec) (P < 0.0001) and those in the 7 segments revealing deteriorated or unchanged wall motion changed from 2.1 +/- 1.0 to -1.7 +/- 0.8 (1/sec) (P < 0.05). In conclusion, strain rate agreed well with assessed wall motion. Strain rate imaging may be a new powerful tool to quantify regional wall contraction. PMID- 12848680 TI - Imagent improves endocardial border delineation, inter-reader agreement, and the accuracy of segmental wall motion assessment. AB - BACKGROUND: This study was conducted to assess the ability of a new echocardiographic contrast agent, Imagent (perflexane lipid microspheres; Alliance Pharmaceutical Corp., San Diego, CA), to improve endocardial border delineation (EBD) and assessment of segmental wall motion (SWM). This was achieved by analysis of inter-reader agreement by echocardiography and comparison with an independent imaging technique, magnetic resonance imaging (MRI). METHODS: Two separate, independent, prospective, randomized, controlled, multicenter trials were conducted at 26 centers and included a total of 409 efficacy evaluable patients. In Study A 206 patients were randomized to receive either Imagent or saline and in Study B, 203 patients received Imagent with a subset imaged by both echocardiography and MRI. All patients were required to have suboptimal baseline images using fundamental imaging. Images were optimized at baseline prior to contrast and the settings maintained post-contrast. Imagent, a suspension of perfluorohexane-filled spheres with flexible lipid shells, was administered as an IV bolus at 0.125 mg/kg body weight. Gated MRI studies were performed within 48 hours of dosing in a subset of 26 subjects. Six expert independent blinded readers reviewed unpaired noncontrast and contrast exams and scored EBD and SWM. Analysis of inter-reader agreement was performed by comparing the SWM score (1 to 5) recorded by each reader pair. In addition, unanimity between readers for SWM was evaluated. For comparison to MRI, the results from echo readers 4, 5, and 6 were each compared with a single independent MRI reader. RESULTS: The patients enrolled in these clinical trials displayed markedly suboptimal images with 49% and 71% (Study A and Study B) of the segments determined by the readers to be suboptimal prior to contrast administration. All readers recorded statistically significant (P < 0.0001) improvement in total EBD scores following the administration of Imagent. Comparison of noncontrast SWM scores for each pair of echo readers resulted in agreement in an average of 39%, of segments in Study A, and 31% of the segments in Study B. Use of Imagent improved agreement in SWM scores to 65% in Study A, and 48% of segments in Study B (P < 0.0001) for all reader pairs in both studies. Reader unanimity in SWM scores increased from 13% to 41% of the segments with the administration of Imagent. Blinded review of the noncontrast echo examinations resulted in agreement with MRI derived SWM scores in 15% of the segments. The administration of Imagent improved this agreement to 47%, of the segments (P < pr = 0.0005 for each blinded reader). CONCLUSIONS: Use of Imagent during echocardiographic imaging improves EBD, providing a significant improvement in inter-reader agreement in SWM evaluation with echo and greater than a threefold improvement in SWM scoring accuracy with MRI. PMID- 12848681 TI - Doppler estimation of filling pressures in a patient with hypertrophic cardiomyopathy. AB - A case of hypertrophic cardiomyopathy is presented with a restrictive mitral filling pattern but predominant systolic pulmonary vein flow velocities. A stepwise Doppler work-up including color M-mode, Doppler tissue imaging of the mitral annulus velocity and loading manipulations could clarify the underlying hemodynamic mechanisms. The Doppler data were correlated with invasive measurements. PMID- 12848682 TI - Post systolic shortening in nonobstructive hypertrophic cardiomyopathy with delayed emptying of the apex: a Doppler flow, tissue Doppler and strain rate imaging case study. AB - We report a case of a 92-year-old woman with syncope and exertional dyspnea, who on echocardiographic examination proved to have hypertrophic cardiomyopathy. Doppler flow revealed delayed emptying of the apex, extending into the early filling phase. Tissue Doppler and strain rate imaging illustrated postsystolic shortening corresponding to the apical ejection, demonstrating that the apical flow was due to wall thickening resulting in active ejection. The imaging methods and mechanics of post-systolic shortening are discussed. PMID- 12848683 TI - Diffuse locally invasive lipomatosis of the pericardium. AB - The chest radiographic, echocardiographic (transthoracic and transesophageal), MRI, CT and cardiac catheterization findings in a 72-year-old patient with extensive pericardial lipomatosis are presented. Diastolic pressures in the left heart were elevated. The massive lipomatous mass was partially resected surgically with good symptomatic relief. Histologically the neoplasm consisted of mature non-malignant adipose tissue. Cushing's Syndrome was absent. PMID- 12848684 TI - Atrial mass and antiphospholipid syndrome. PMID- 12848685 TI - Bicuspid aortic valves in monozygotic twins. AB - Identical bicuspid valve anomalies were found in monozygotic twins. Screening echocardiography should be considered for first-degree relatives of patients with bicuspid aortic valve. PMID- 12848686 TI - Drainage of the inferior vena cava to the left atrium. AB - Drainage of the inferior vena cava to the left atrium is an extremely unusual congenital heart disease. We describe a 54-year-old woman, in whom the diagnosis was suggested by transthoracic echocardiography, and then confirmed by a transesophageal exam and magnetic resonance imaging, which also revealed an associated secundum atrial septal defect. Surgical management involved reconstruction of the interatrial septum to include the inferior vena cava in the right atrium. The few previously reported cases in the literature are reviewed. PMID- 12848687 TI - Asymptomatic isolated congenital left ventricular muscular diverticulum in an adult: a case report. AB - Congenital ventricular diverticulum is a very rare malformation in adults. We describe a 21-year-old male with a congenital muscular left ventricular diverticulum in the inferior wall. The lesion was suspected on two-dimensional transthoracic echocardiography; transesophageal echocardiography allowed clear detection of the diverticulum as well of mild mitral valve prolapse. The diagnosis was confirmed by cardiac catheterization. There were no other thoracoabdominal or cardiac anomalies, the patient was asymptomatic, and surgery was not deemed necessary. PMID- 12848688 TI - Transthoracic echocardiographic demonstration of septal perforator coronary arteries-left ventricle fistula. AB - We present an adult patient status post septal myectomy in whom transthoracic two dimensional echocardiography demonstrated septal perforator coronary arteries opening directly into the left ventricle. In some views this finding mimicked aortic regurgitation. PMID- 12848689 TI - Cisplatin induced localized aortic thrombus. PMID- 12848690 TI - Massive metastatic intracardiac lymphoma presenting with complete heart block with resolution following chemotherapy. PMID- 12848691 TI - Usefulness of transesophageal three-dimensional echocardiography in the identification of individual segment/scallop prolapse of the mitral valve. AB - We evaluated the potential usefulness of three-dimensional (3D) transesophageal echocardiography (TEE) in assessing individual scallop/segment prolapse in 36 adult patients with mitral valve prolapse (MVP) undergoing surgical correction. Intraoperative 3D TEE correctly identified the location of scallop/segment prolapse in 34 of 36 patients (94%). However, in 6 of these patients 3D TEE images revealed more scallops or segments with prolapse than the surgeon noted intraoperatively. Prolapse of these areas was less prominent and this could possibly explain the lack of correlation with the surgical findings in these patients. In another 2 patients areas of prolapse seen by the surgeon were missed by 3D TEE because some of those scallops/segments could not be well imaged due to image "drop out" and artifacts. Thus, perfect correlation between 3D TEE and surgery was noted in 28 of 36 (78%) patients. Noncoaptation of the MV was also identified in 2 patients. The prolapsed area of posterior (n = 28 observations) and anterior (n = 9 observations) MV leaflets ranged from 1 cm2 to 9 cm2 (mean 3.50 cm2+/- 2.14) and 1.20 cm2 to 5.99 cm2 (mean 3.21 cm2+/- 1.33), respectively. Interobserver and intraobserver agreement for location and area of MVP was excellent (r = 0.97 and r = 0.99, respectively; all P values are <0.0001). In conclusion, 3D TEE is useful in identifying the location of MVP. It may also be potentially useful in assessing the extent of individual scallop/segment prolapse and identifying sites of MV noncoaptation. This information could aid the surgeon in deciding the extent of MV resection. PMID- 12848692 TI - Myocardial performance index (Tei index) does not reflect long-term changes in left ventricular function after acute myocardial infarction. AB - AIMS: To evaluate whether changes in myocardial performance index (MPI or Tei index) were related to changes in other Doppler echocardiographic parameters after acute myocardial infarction, or had any independent prognostic impact in a 2-year observational study. METHODS AND RESULTS: Seventy-one patients with acute myocardial infarction without heart failure were examined at baseline, 3 months, and 2 years. MPI was significantly related to end-diastolic and end-systolic volume indexes, ejection fraction, maximal velocity, and time velocity integral of early mitral filling wave at 3 months and 2 years. MPI did not contribute significantly to the prediction of any changes in the measures of diastolic or systolic function at 3 months or 2 years. Baseline MPI was significantly higher in patients who later developed heart failure(0.55 +/- 0.16)than in other patients(0.43 +/- 0.13, P = 0.006), but had no independent predictive power for the development of heart failure or death relative to end-systolic volume index and deceleration time of early mitral filling wave. CONCLUSION: MPI did not accurately reflect changes in Doppler and two-dimensional echocardiographic measures of diastolic or systolic function during a 2-year follow-up after acute myocardial infarction, and did not have any independent prognostic impact. PMID- 12848693 TI - Impact of left ventricular function on the pulmonary vein Doppler spectrum: nonsimultaneous assessment with load-insensitive indices. AB - Pulmonary vein Doppler spectrum is highly load-dependent and thus has been used to estimate left ventricular (LV) filling pressure. However, the impact of LV function on pulmonary vein Doppler spectrum remains obscure because only load sensitive indices were studied previously. In the present study, measurements of the pulmonary vein Doppler spectrum were correlated with load-insensitive LV systolic (end-systolic elastance [Ees]) and diastolic (relaxation time constant [tau] and beta coefficient of the end-diastolic pressure volume relationship) function indices obtained from an invasive catheterization study nonsimultaneously. The peak velocity, velocity time integral, and duration of systolic forward spectrum were significantly correlated with Ees (r = 0.35, r = 0.36, and r = 0.41, respectively;P < 0.05). The pulmonary vein diastolic velocity time integral (PVDVTI) and duration of the diastolic forward spectrum were significantly correlated with Ees (r = 0.51 and r = 0.57, respectively;P < 0.01). PVDVTI was correlated with tau and the end-diastolic pressure-volume relationship (EDPVR) (r = 0.42 and r = 0.40 respectively,P < 0.05). On the other hand, the systolic fraction of the forward spectrum was significantly correlated with ejection fraction (for peak velocity,r = 0.63, P < 0.01; for velocity time integral,r = 0.37, P < 0.05) but not with Ees, and the diastolic fraction of the forward spectrum was significantly correlated with minimum pressure derivative over time (for peak velocity,r = 0.48, P < 0.05; for velocity time integral,r = 0.44, P < 0.05, respectively) but not with tau or EDPVR. In summary, the systolic and diastolic components of the pulmonary vein Doppler spectrum are affected variably by LV systolic and diastolic function, independent of the loading condition. The systolic and diastolic fraction of pulmonary vein Doppler spectrum appears to depend more on the loading condition than the LV systolic or diastolic function. PMID- 12848694 TI - Assessment of left and right ventricular systolic and diastolic synchronicity in normal subjects by tissue Doppler echocardiography and the effects of age and heart rate. AB - OBJECTIVES: This study examined inter- and intraventricular synchronicity in systole and diastole by tissue Doppler imaging (TDI), and investigated if these parameters and the regional velocities were affected by age and heart rate. METHODS: TDI was performed in 106 normal subjects (64.3 +/- 9.5 years, 60% male) using three apical views and a six-basal, six mid-segmental model. The regional parameters measured off line in both ventricles included peak isovolumic contraction velocity IVC(M), peak sustained systolic velocity (SM), peak early diastolic velocity (EM), peak late diastolic velocity (AM), and the E/AM ratio, as well as their time to these peak velocities: T(IVC), T(S), T(E), and T(A). RESULTS: The systole and diastole within the left ventricle (LV) was highly synchronized without difference in T (IVC), TS, TE, and TA. However, the right ventricle (RV) was about 20 msec later than the LV for T(IVC) and TS. For regional velocities, IVC(M), S(M), E(M), and A(M) were significantly higher in basal than mid-segments (all P < 0.001). In the base of the LV, SM, and EM were the highest at the lateral segment and the lowest at the anterolateral segment. Age and heart rate did not affect systolic velocities or the timing of events. In diastole, age correlated negatively with EM(r =-0.36 to -0.48, P 0.85) provided that the repeat reading was taken at least 15 min after the initial reading. The examiner found the equipment easy to use and taking measurement was readily acceptable by the sample group. It was concluded that Periotest could be utilised in traumatology, although the clinician needs to take into account the limitations of the device and the difficulty in interpreting the results. PMID- 12848716 TI - The Periotest in traumatology. Part II. The Periotest as a special test for assessing the periodontal status of teeth in children that have suffered trauma. AB - The research was carried out on the upper permanent incisor teeth of 101 children aged 8-12 years attending the trauma clinic at the University Dental Hospital, Manchester, UK. The double blind, cross-over study involved Periotest readings being taken by one examiner and traditional special tests being carried out by another examiner. The traditional special tests chosen were mobility testing, percussion sounds and radiographs. The Periotest readings and special tests were then repeated at the child's recall appointment. The results revealed that there was a significant difference (P < 0.001) between the number of abnormalities detected by the Periotest and those detected by the traditional special tests. There was also found to be a significant difference (P < 0.001) in the detection of undesirable sequelae between the Periotest and the traditional special tests. PMID- 12848717 TI - Preservation of ridge dimensions following grafting with coral granules of 48 post-traumatic and post-extraction dento-alveolar defects. AB - This prospective clinical analysis reports on the use of coral granules in alveolar ridge preservation procedures in a population of young, growing patients. The sample consisted of 21 patients, 12 females and 9 males, with a mean age of 13.6 years. These 21 patients had 48 dento-alveolar defects suitable for augmentation with coral granules, and were followed clinically and radiographically for 3-7 years after augmentation. There were two areas of augmentation: 17 defects in the anterior maxilla resulted from traumatic tooth loss, and 31 defects in the posterior maxilla and mandible resulted from the extraction of ankylosed retained primary molars with no permanent succedaneous teeth. Between 1-2 ml of coral granules were implanted into the alveolar bone defects left by the extraction of teeth in both the areas. This was in order to preserve the remaining edentulous ridge from further alveolar ridge resorption. The goal of the procedure was to preserve the alveolus so that in the future, a dental implant could be placed to replace the missing tooth, after jaw growth had stopped, without the need for a bone graft. The coral granules appeared to be totally replaced by the host bone on follow-up clinical and radiographic examinations. The two areas of the jaws behaved quite differently. In the anterior maxilla, where tooth loss was secondary to trauma, the coral granules restored the alveolar ridges temporarily. However, over the years of follow-up in this study, the coral granules failed to provide sufficient bone to support the placement of a dental implant without using a bone graft in 14 of the 17 defects or 82.4% of sites. In the posterior maxilla and mandible, where tooth loss was due to the elective removal of ankylosed primary molars, 29 of the 31 defects or 93.5% of sites were successful as they were able to support the placement of an osseo-integrated dental implant without the use of a bone graft. The alveolar sparing technique was more successful in maintaining an alveolar ridge sufficient for the placement of a dental implant without bone grafting in the posterior maxilla and mandible, where tooth loss was secondary to the elective removal of ankylosed deciduous molars than in the anterior maxilla, where tooth loss was secondary to trauma. Coral granules seem to be more suitable in the posterior maxilla and mandible where there were ankylosed deciduous teeth and congenitally absent permanent teeth than in the traumatized anterior maxilla. In successful sites, coral granules can spare the alveolus from residual ridge atrophy or resorption, obviating the need for a bone graft. This reduces patient morbidity, as a second surgical donor site is avoided because bone graft harvesting is made unnecessary. PMID- 12848718 TI - Endodontic retreatment of an autotransplanted lower first premolar: a case report. AB - A 24-year-old female with no contributory medical history had been treated by orthodontic alignment and transplantation of the lower left first premolar to the upper left first premolar site to correct for congenitally missing upper premolars. Subsequently, the transplanted premolar was treated by conventional endodontics and apical surgery for persistent periapical periodontitis over a span of 12 years. A combination of conventional endodontic retreatment and periapical surgery resulted in periapical healing. This case outlines the need for careful preoperative case assessment. PMID- 12848719 TI - Intentional replantation of a mandibular second molar with long-term follow-up: report of a case. AB - A case is reported of a mandibular second molar that underwent non-surgical endodontic treatment and intentional replantation in an effort to relieve continued symptoms. A 15.5-year follow-up clinical examination revealed the patient to be asymptomatic, the tooth to be still functional, and a recall film showed no evidence of root resorption. The indications for and limitations of intentional replantation as well as recommended literature on the subject are discussed. PMID- 12848722 TI - Antimicrobial susceptibility of Bacteroides fragilis group isolates in Europe. AB - OBJECTIVE: To evaluate the activity of old and newer antianaerobic drugs against clinical isolates of Bacteroides fragilis group strains from different parts of Europe. METHODS: Bacteroides fragilis group isolates from 37 laboratories in 19 countries were biochemically characterized. The MICs of seven antimicrobial agents were determined by the agar dilution method as recommended by the NCCLS. Production of beta-lactamase was detected by nitrocefin. RESULTS: There were 1284 B. fragilis group isolates included in the study. Abdominal infections and wounds were the most common sources of isolation and B. fragilis was the dominating species. Ninety-nine percent of the strains were resistant to ampicillin (breakpoint 2 mg/L), 6% to cefoxitin (64 mg/L), 15% to clindamycin (8 mg/L) and 9% to moxifloxacin (8 mg/L). Less than 1% were resistant to imipenem (16 mg/L), piperacillin-tazobactam (128 mg/L) and metronidazole (32 mg/L). Ninety-six percent of the isolates were beta-lactamase producers. CONCLUSIONS: Antimicrobial resistance among the B. fragilis group is increasing. PMID- 12848721 TI - Multicenter evaluation of the reproducibility of the proposed antifungal susceptibility testing method for fermentative yeasts of the Antifungal Susceptibility Testing Subcommittee of the European Committee on Antimicrobial Susceptibility Testing (AFST-EUCAST). AB - OBJECTIVE: To evaluate the intra- and inter-laboratory reproducibility of a new standard for susceptibility testing of fermentative yeasts. This standard is based on the M27-A procedure of the National Committee for Clinical Laboratory Standards (NCCLS), but incorporates several modifications, including spectrophotometric growth-dependent endpoint reading. METHODS: Nine laboratories participated in the study. Common material lots were used to test six Candida species (one each of C. albicans, C. tropicalis, C. parapsilosis, C. glabrata, C. krusei, and C. lusitaniae), and two quality control strains (C. krusei ATCC6258 and C. parapsilosis ATCC22019). Triplicate testing on three separate days was performed in microtiter format with RPMI-2% glucose, pH 7.0. Flucytosine, fluconazole and itraconazole were tested. In total, 3888 MIC values were included in the analyses. Reproducibility was calculated by means of agreement (percentage of MICs within one two-fold dilution of the mode) and intraclass correlation coefficient (ICC, maximum value of 1). RESULTS: The average intra-laboratory agreements were 99% and 96% after 24 h and 48 h of incubation, respectively, with ICCs of 0.98 and 0.97 (P < 0.05). Two strains exhibiting a trailing effect showed intra-laboratory agreement of 92% and ICCs of < 0.91 at 48 h. The inter laboratory agreement was 94% and 88% after 24 h and 48 h, respectively, with ICCs of 0.93 and 0.91 (P < 0.05). Lower values of agreement and ICCs were obtained for strains exhibiting trailing after 48 h of incubation. Itraconazole yielded the lowest values of reproducibility. CONCLUSION: The new procedure of EUCAST for antifungal susceptibility testing is a reproducible method within and between laboratories and offers several advantages over the NCCLS approved method. PMID- 12848723 TI - Non-invasive techniques for the diagnosis of Helicobacter pylori infection. AB - Helicobacter pylori infection can be diagnosed by invasive techniques requiring endoscopy and biopsy (histologic examination, culture, polymerase chain reaction), and non-invasive techniques (serology, urea breath test, urine or blood, detection of H. pylori antigen in stool specimen). However, recent studies have demonstrated that a strategy of 'testing and treating' for H. pylori in uninvestigated, young (<50 years), dyspeptic patients in primary care is safe and reduces the need for endoscopy. Indeed, a number of clinical guidelines recommend non-invasive testing in dyspeptic patients followed by treatment of H. pylori in primary care based on clinical and economic analyses. Several non-invasive tests are currently available on the market. The choice depends on the clinical circumstances, the likelihood ratio of positive and negative tests, the cost effectiveness of the testing strategy, and, finally, the availability of the tests. Nevertheless, two non-invasive tests are commonly used: the urea breath test, and the stool antigen test. PMID- 12848724 TI - Current strategies to avoid misdiagnosis of malaria. AB - Malaria remains the most important parasitic disease, and tens of thousands of cases are imported into non-endemic countries annually. However, any single institution may see only a very few cases-this is probably the reason why laboratory and clinical misdiagnosis may not be uncommon. In the laboratory, unfamiliarity with microscopic diagnosis may be the main reason, considering the large number of laboratory staff who provide on-call services, often without expert help at hand, as well as the difficulty in detecting cases with low-level parasitemia. Staff should therefore be provided with continuing microscopic training to maintain proficiency. The complementary use of immunochromatographic rapid detection tests (RDTs) may be useful, especially during on-call hours, although, in order to ensure correct interpretation, their inherent limitations have to be well known. Diagnosis based on the polymerase chain reaction is still unsuitable for routine use, due to its long turnaround time, its cost, and its unavailability outside regular hours, although it may be helpful in selected cases. Once the alert clinician has considered the possibility of malaria, and suspicion continues to be high, malaria can be excluded by repeat smears or RDTs. However, the absence of clinical suspicion may not be infrequent, and may have more serious consequences. Depending on the local number of malaria cases seen, laboratory staff should have a low threshold for the decision to perform unsolicited malaria diagnostic tests on suspicious samples, especially if other laboratory tests are abnormal (e.g. thrombocytopenia, presence of atypical lymphocytes, or raised lactate dehydrogenase). The detection of intraleukocytic hemozoin during automated full blood counts is a promising new way to avoid misdiagnosis of clinically unsuspected malaria. PMID- 12848725 TI - Molecular diagnostics in clinical parasitology and mycology: limits of the current polymerase chain reaction (PCR) assays and interest of the real-time PCR assays. AB - Polymerase chain reaction (PCR) represents a major breakthrough for the diagnosis of infectious diseases. However, the absence of standardized kits for commercially unattractive targets, such as most of the parasites and the fungi, has led to the development of numerous in-house PCR assays. The performances reported, both for the sensitivity and the specificity of these assays are very divergent. For instance, for the antenatal diagnosis of toxoplasmosis, the sensitivity is either 97.4%, or 64%. For the diagnosis of toxoplasmosis in HIV positive patients, the PCR on blood is either of limited value with a sensitivity of 13% or of excellent yield with a sensitivity of 87.5%. Similar results are reported for the diagnosis of invasive aspergillosis in bone-marrow-transplant recipients. The patients and the clinical specimens tested are often different. This can explain some of the discrepancies. However, when performed, the quality controls on identical specimens show different results depending on the laboratories. An analysis of the PCR techniques used shows that the control of false positive results as a result of carry-over and false negative results owing to PCR inhibitors is far from being systematic. These shortcomings of 'classical' PCR should be solved when real-time PCR assays are developed, leading to some standardization. Automated DNA extraction should also be useful to achieve this goal. Comparison between laboratories should then be possible and regular quality controls will be necessary to ensure the reliability of real-time PCR assays. PMID- 12848726 TI - Mechanism of quinolone resistance in anaerobic bacteria. AB - Several recently developed quinolones have excellent activity against a broad range of aerobic and anaerobic bacteria and are thus potential drugs for the treatment of serious anaerobic and mixed infections. Resistance to quinolones is increasing worldwide, but is still relatively infrequent among anaerobes. Two main mechanisms, alteration of target enzymes (gyrase and topoisomerase IV) caused by chromosomal mutations in encoding genes, or reduced intracellular accumulation due to increased efflux of the drug, are associated with quinolone resistance. These mechanisms have also been found in anaerobic species. High level resistance to the newer broad-spectrum quinolones often requires stepwise mutations in target genes. The increasing emergence of resistance among anaerobes may be a consequence of previous widespread use of quinolones, which may have enriched first-step mutants in the intestinal tract. Quinolone resistance in the Bacteroides fragilis group strains is strongly correlated with amino acid substitutions at positions 82 and 86 in GyrA (equivalent to positions 83 and 87 of Escherichia coli). Several studies have indicated that B. fragilis group strains possess efflux pump systems that actively expel quinolones, leading to resistance. DNA gyrase seems also to be the primary target for quinolones in Clostridium difficile, since amino acid substitutions in GyrA and GyrB have been detected in resistant strains. To what extent other mechanisms, such as mutational events in other target genes or alterations in outer-membrane proteins, contribute to resistance among anaerobes needs to be further investigated. PMID- 12848727 TI - Mycobacterial infection in a series of 1261 renal transplant recipients. AB - OBJECTIVE: To describe the incidence and clinical characteristics of mycobacterial infection in renal transplant recipients. METHODS: We retrospectively analyzed the cases of mycobacterial infection in a series of 1261 renal transplants carried out in our Unit of Renal Transplantation from 1980 to 2000. Demographic parameters and clinical antecedents such as age, cause of end stage renal disease, time of follow-up of the graft, previous renal function and type of immunosuppression were considered. Moreover, the clinical onset, diagnostic tools, treatment policy and evolution were studied. The pathogenesis of the different types of mycobacteria isolated was also analyzed. Diagnosis was made with the Ziehl-Neelsen staining method. Culture was performed by the conventional Lowenstein-Jensen method and the Bactec-460 radiometric method. RESULTS: We found mycobacterial infection in 27 patients (2.1%), due to Mycobacterium tuberculosis in 20 cases, M. kansasii in five patients, and M. fortuitum in two patients. The mean elapsed time from the renal transplant was 20.5 months; the infection appeared in 18 patients during the first eight months after transplantation. The clinical onset was pulmonary infection in 17 cases (12 M. tuberculosis and five M. kansasii); five had urinary symptoms (three M. tuberculosis and two M. fortuitum); three cases of M. tuberculosis infection had abdominal symptoms; another one began with a perineal tuberculous abscess; the rest of the patients were asymptomatic. The types of specimen on which microbiological identification was carried out were, in decreasing order: sputum and/or bronchial washing/pleural aspiration, urine, feces, gastric and peritoneal fluids, bone marrow and blood. The first-line drug isoniazid had the highest resistance index in the susceptibility test. Clinical dissemination was observed in eight patients, four of whom died. Another three patients had a significant impairment in renal function, and in one of these patients an allograft nephrectomy was necessary due to a severe septic syndrome. CONCLUSIONS: Mycobacterial infection, mainly by M. tuberculosis, has an important impact on kidney transplant recipients, particularly during the first year after surgery. Diagnosis often presents some difficulties, and a delay in treatment represents a determinant factor for the evolution, with a risk of death or permanent damage in renal function. Therefore, early diagnosis is mandatory. When the Mantoux reaction is positive, antituberculous prophylaxis seems advisable. PMID- 12848728 TI - Antecedent use of fluoroquinolones is associated with resistance to moxifloxacin in Clostridium difficile. AB - OBJECTIVE: Moxifloxacin is characterized by high activity against Gram-positive cocci and some Gram-positive and -negative anaerobes, including Clostridium difficile. This study investigates the role of prior quinolone use in relation to patterns of susceptibility of C. difficile to moxifloxacin. METHODS: Sixty-three clinical isolates of C. difficile were investigated for toxigenicity, susceptibility to moxifloxacin, and mutations in the DNA gyrase gene. The medical histories for 50 of these patients were available and used to identify previous fluoroquinolone use. RESULTS: Thirty-three (52.4%) strains showed resistance to moxifloxacin (MICs > or = 16 mg/L). All moxifloxacin-resistant strains harbored a mutation at amino acid codon Ser-83 of gyrA. Forty-five isolates (71.4%) were toxigenic; all moxifloxacin-resistant strains were in this group. Resistance to moxifloxacin was associated with prior use of fluoroquinolones (P-value 0.009, chi-square). CONCLUSIONS: Although the use of moxifloxacin to treat C. difficile associated diarrhea is not likely to be common, these data show a relationship between antecedent fluoroquinolone use and resistance to moxifloxacin in C. difficile isolates, and raise questions regarding selection pressure for resistance placed on colonizing bacteria exposed to fluoroquinolones. Mutations in gyrA are involved in moxifloxacin resistance. PMID- 12848729 TI - Changing epidemiology and predictors of mortality in patients with spontaneous bacterial peritonitis at a liver transplant unit. AB - OBJECTIVES: To determine whether antimicrobial resistance in pathogens and outcome in patients with spontaneous bacterial peritonitis (SBP) has evolved over time. METHODS: Sixty-one consecutive episodes of SBP were studied in patients with end-stage liver disease undergoing evaluation for liver transplantation between 1991 and 2001. Patients were dichotomized into a cohort between 1991 and 1995 (the earlier cohort) and 1996-2001 (the later cohort). RESULTS: Overall, 19% of all bacteria were multiply-antibiotic resistant. The frequency of multiple antibiotic resistance in bacteria increased from 8.3% to 38.5% in the earlier as compared to the later cohort (P = 0.07). Overall, mortality at 30 days in the study patients was 26% and had remained unchanged between the two cohorts. The mortality rate was significantly higher in patients with multiply-antibiotic resistant bacteria than in those with other bacteria (P = 0.045). However, the Child-Pugh score (P = 0.003), and renal failure (P = 0.04) were the only independently significant predictors of mortality in patients with SBP. CONCLUSIONS: Mortality in patients with end-stage liver disease who developed SBP has remained unchanged over the last decade. Although multiple-antibiotic resistance in bacteria causing SBP has increased over time, the severity of hepatic and renal dysfunction were the predominant determinants of outcome in these patients. PMID- 12848730 TI - Clinical microbiological case: a necrotic skin lesion in a patient with renal failure. PMID- 12848731 TI - Remote abscess formation during interferon-alpha therapy for viral hepatitis. AB - Abscess formation at the injection site is an unusual infectious complication of interferon-alpha (IFN-alpha) treatment of chronic hepatitis C virus (HCV) infection, but remote abscess formation during IFN-alpha therapy is very rare. In the present communication, we report three cases of remote abscess formation detected among 68 patients with chronic viral hepatitis treated with IFN-alpha, and review the pertinent English literature. We believe that, as fever and constitutional symptoms are common side effects of IFN-alpha treatment, a high index of suspicion is indicated to exclude abscess formation in cases of unexplained fever during IFN-alpha therapy. PMID- 12848732 TI - Validation of commercial dry-form broth microdilution panels for susceptibility testing of AZD2563, a new long-acting oxazolidinone. AB - The MIC results using a dry-form broth microdilution panel (TREK Diagnostic/Sensititre, Westlake, OH, USA) were validated for AZD2563, a novel oxazolidinone compound. In comparision studies against reference frozen-form panels, the commercial MIC results were the same as the reference calue for 82.7% of organisms and all results were within +/- one log2 dilution. Using 462 organisms, most from three genus groups (enterococci, staphylococci, streptococci), test results indicate that Sensititre MIC values were comparable to the reference test and can be utilized in clinical trials of for routine laboratory use when testing AZD2563 and linezolid, the drug class comparator. PMID- 12848733 TI - Multiresistant Acinetobacter baumannii isolates in intensive care units in Greece. AB - One hundred and twenty-one clinical isolates of Acinetobacter baumannii recovered from the intensive care units (ICUs) of nine tertiary-care hospitals in Athens, Greece were studied in order to determine whether the increasing appearance of resistant acinetobacters is due to the spread of epidemic strains. The majority of the isolates exhibited resistance to ampicillin-sulbactam, and the most common antibiotic resistance profiles comprised resistance to nine and eight of the 11 potentially active antibiotics tested, respectively. Pulsed-field gel electrophoresis showed that 68% of the isolates, recovered from all ICUs, belonged to two clonal groups, indicating inter-hospital dissemination of multiresistant A. baumannii in our region. PMID- 12848734 TI - Transmission of Streptococcus pyogenes causing successive infections in a family. AB - The objective of this study was to determine the characteristics of Streptococcus pyogenes isolated during a 10-month period from members of a family with infections and asymptomatic carriage. T-serotyping and pulsed-field gel electrophoresis confirmed that distinct GAS clones were introduced into the family over a short period of time. PMID- 12848735 TI - A parent as a vector of Salmonella brandenburg nosocomial infection in a neonatal intensive care unit. AB - A newborn baby was admitted to the Neonatal Intensive Care Unit (NICU) of St Germain en Laye Hospital (France) because of premature birth. On day 12, he contracted gastroenteritis due to Salmonella brandenbourg. The salmonellosis led to a septic shock syndrome with a brief cardiopulmonary arrest. He was treated with intravenous ceftriaxone and gentamicin, and the evolution was favorable. Microbiological investigations revealed that the mother was the vector for this nosocomial infection. S. brandenbourg was isolated from the feces of the baby, despite recent recommendations on managing stool specimens from patients hospitalized for more than three days: according to these recommendations, these stools should be processed for viruses and Clostridium difficile toxin only. PMID- 12848736 TI - The activity of meropenem and comparators against Acinetobacter strains isolated from European hospitals, 1997-2000. AB - In vitro susceptibilities to meropenem and comparators of Acinetobacter strains isolated from serious infections in 37 European hospital centers participating in the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) Program (1997-2000) were tested. There were 635 Acinetobacter strains collected: 490 A. baumannii; 51 A. calcoaceticus var. lwoffii; and 94 other Acinetobacter strains. Overall, meropenem and imipenem were the most effective agents tested. Resistance to the antimicrobials was: 14%, meropenem; 16%, imipenem; 39%, piperacillin tazobactam; 41%, tobramycin; 45%, ceftazidime; and 53%, ciprofloxacin. Thus, the carbapenems have useful activity against Acinetobacter spp. and represent a viable choice for treating infections caused by these organisms. PMID- 12848737 TI - A fatal case of Vibrio vulnificus meningoencephalitis. AB - The objective of this paper is to report a rare case of Vibrio vulnificus presenting as meningoencephalitis without a wound infection. Vibrio vulnificus is capable of causing severe and often fatal infections in susceptible individuals. It commonly causes necrotizing wound infections, primary septicemia, and gastroenteritis. A 69-year-old man had meningoencephalitis with lesion on the red nucleus, substantia nigra, basal ganglia, and dentate nucleus as the initial clinical manifestation of a V. vulnificus infection. This is the first case of V. vulnificus infection in which MRI demonstrated the involvement of deep nuclei of the brain. PMID- 12848738 TI - Undiagnosed systemic lupus erythematosus presenting with salmonella bacteremia: a case report and mini-review. PMID- 12848739 TI - Comparative evaluation of test assays for detection of methicillin resistance in Staphylococcus aureus. PMID- 12848740 TI - A case of endocarditis due to Granulicatella adiacens. PMID- 12848741 TI - Absence of risk factors for community MRSA. PMID- 12848745 TI - Intravenous immunoglobulins in infectious diseases: where do we stand? AB - Intravenous immunoglobulins (IVIg) are therapeutic preparations of normal human IgG that have been used for more than 20 years for substitutive therapy in patients with primary antibody deficiencies. Recent studies pointed out the need to obtain normal residual levels of IgG (i.e. 8 g/L) in order to reduce the number and severity of bacterial infections in these patients. The IVIg are also prescribed for the substitutive therapy of secondary immunodeficiencies such as chronic lymphoid leukemia and multiple myeloma with hypogammaglobulinemia and severe and/or recurrent infections, and human immunodeficiency virus (HIV) infected children with recurrent bacterial infections before the era of highly active antiretroviral agents. However, in the latter situation, no recent study has evaluated IVIg therapy in acquired immunodeficiency syndrome (AIDS) children receiving highly active antiretroviral agents (HAART), and the use of IVIg must probably be restricted to the currently rare clinical situation in Western Europe of children with AIDS who develop recurrent infections despite the administration of HAART and prophylactic cotrimoxazole. IVIg have also been reported to prevent infections, interstitial pneumonia and graft-vs. host disease during the first 90 days post-transplant in allogeneic bone-marrow transplant recipients. However, this result was not confirmed by two recent studies and IVIg therapy should probably only be proposed for a subgroup of bone-marrow allografted patients such as those with hypogammaglobulinemia and sepsis. With the exception of erythrovirus B19 infection with erythroblastopenia, no clear benefit of IVIg therapy has been reported for the curative management of other infectious diseases. PMID- 12848746 TI - Is there a rationale for the continuous infusion of cefepime? A multidisciplinary approach. AB - This review is the fruit of multidisciplinary discussions concerning the continuous administration of beta-lactams, with a special focus on cefepime. Pooling of the analyses and viewpoints of all members of the group, based on a review of the literature on this subject, has made it possible to test the hypothesis concerning the applicability of this method of administering cefepime. Cefepime is a cephalosporin for injection which exhibits a broader spectrum of activity than that of older, third-generation cephalosporins for injection (cefotaxime, ceftriaxone, ceftazidime). The specific activity of cefepime is based on its more rapid penetration (probably due to its zwitterionic structure, this molecule being both positively and negatively charged) through the outer membrane of Gram-negative bacteria, its greater affinity for penicillin-binding proteins, its weak affinity for beta-lactamases, and its stability versus certain beta-lactamases, particularly derepressed cephalosporinases. The stability of cefepime in various solutions intended for parenteral administration has been studied, and the results obtained demonstrated the good compatibility of cefepime with these different solutions. These results thus permit the administration of cefepime in a continuous infusion over a 24-h period, using two consecutive syringes. PMID- 12848747 TI - Rapid detection of resistance in Mycobacterium tuberculosis: a review discussing molecular approaches. AB - The last few years have seen the development of several molecular designs to search for mutations encoding resistance to antituberculous drugs in Mycobacterium tuberculosis. Most of these are highly efficient for RIF-r detection and are well adapted to search for the most relevant INH-R mutations. In this review, these new molecular approaches are explained and are presented according to the molecular strategies on which they are based. In this sense, techniques based on DNA-sequencing, electrophoresis and hybridization are reviewed and the newer designs based on real-time PCR and microarrays are also included. Molecular methods are sure to transform standard approaches to the issue of resistance in the mycobacteriology laboratory. This will allow laboratories to speed up the performance of resistance assays and provide access to essential information for highly refined detection, follow-up and management of antibiotic resistance in M. tuberculosis. PMID- 12848748 TI - Regional dissemination of Salmonella enterica serovar Enteritidis is season dependent. AB - OBJECTIVE: To carry out epidemiological typing of clinical isolates of Salmonella enterica serovar Enteritidis by pulsed-field gel electrophoresis (PFGE), random amplified polymorphic DNA (RAPD) and analysis of their antibiotic resistance. METHODS: Over a 12-month period, 44 Salmonella Enteritidis isolates, recovered from 40 patients admitted to the University Hospital Center of Amiens, France and from three outpatients, were characterized by the analysis of phenotypic and genotypic traits and clinical data from medical reports. RESULTS: Forty nontyphoidal salmonellosis episodes were diagnosed in hospitalized patients (34 episodes of gastroenteritis, two episodes of bacteremia not affecting other organs, one episodes of bacteremia plus urinary infection, one episodes of bacteremia plus gastroenteritis, one episodes of chronic colitis plus gastroenteritis and one episode of peritonitis), and three carriers were observed in outpatients. By means of PFGE, RAPD and antibiotic susceptibility patterns 44 isolates were subdivided into 16 clonally related groups. Two of them were predominantly implicated in the course of these infections, being responsible for two successive waves of infection, while the others were encountered sporadically. PMID- 12848749 TI - Development of resistance in Pseudomonas aeruginosa obtained from patients with cystic fibrosis at different times. AB - OBJECTIVE: Determination of the extent of changes in quantitative resistance in Pseudomonas aeruginosa isolates from patients with cystic fibrosis over a period of approximately 2 years. METHODS: Three hundred and ninety nine isolates of P. aeruginosa collected from 34 pediatric patients in the period between April 1994 and April 1996 were investigated. During the 2 years the children were treated with a combination of a betalactam and an aminoglycoside, approximately every 3 months. In between they received ciprofloxacin orally, when required. The minimal inhibitory concentrations (MICs) of 38 clones of P. aeruginosa defined by different patterns in macrorestriction analysis (pulse field gel electrophoresis, PFGE) were established for 12 antibiotics: gentamicin, amikacin, tobramycin, ciprofloxacin, levofloxacin, moxifloxacin, trovafloxacin, imipenem, meropenem, ceftazidime, cefepime, and piperacillin by means of broth microdilution tests according to DIN 58940. RESULTS: Twenty-four of the 38 clones developed increased MIC values during the time of observation especially for aminoglycosides and quinolones. Comparatively less affected were ceftazidime, imipenem and meropenem. An association between the number of the intravenous treatment courses and the increase of the MIC values could not be verified. CONCLUSIONS: A trend towards an increase of the MICs against antipseudomonal agents was observed over a limited period of time. It is necessary to prevent this development possibly by employing suitable combinations of antibiotics and the introduction of new substances. PMID- 12848750 TI - A series of infections due to Capnocytophaga spp in immunosuppressed and immunocompetent patients. AB - OBJECTIVE: To investigate the epidemiology, microbiology and outcome of infections caused by Capnocytophaga spp. at a single center. METHODS: We report on ten documented infectious episodes caused by Capnocytophaga observed between 1994 and 1999 at the Innsbruck University Hospital. RESULTS: In seven of ten patients, Capnocytophaga septicemia was diagnosed during periods of neutropenia. In contrast, the remaining three patients had normal white blood cell counts when acquiring Capnocytophaga septicemia (one) and pleural empyema (two). Blood cultures containing long, slender, Gram-negative rods, which grew slowly under anaerobic conditions and lacked susceptibility to metronidazole, were subcultivated in a CO2-enriched atmosphere (5%). Subcultivation yielded Capnocytophaga in all ten cases within 2-12 days. The patients were then placed on appropriate antibiotic therapy, with or without additional surgical intervention, and the organism was eradicated. CONCLUSION: Identification of Capnocytophaga facilitates appropriate, and in most cases effective, antimicrobial therapy. PMID- 12848751 TI - Different trends in antibiotic resistance rates at a university teaching hospital. AB - OBJECTIVES: To investigate long-term trends in antibiotic resistance of common bacterial species isolated at a university hospital and in its intensive care units (ICUs). METHODS: Levels of antibiotic resistance of common bacterial pathogens were investigated at the Karolinska Hospital during the 12-year period 1988-99. Resistance rates were analyzed for the entire hospital, as well as for ICUs combined. RESULTS: At the Karolinska Hospital, we found increased ciprofloxacin resistance among Escherichia coli isolates, from 0% in 1991 to 11% in 1999. In the ICUs, the corresponding increase was from 0% to 4.8% during the same period. Co-trimoxazole resistance levels increased from 7.5% to 14%, with lower levels for the ICUs. For ampicillin, cefuroxime, and gentamicin, the levels of resistance were similar in the whole hospital and in the ICUs. Among Pseudomonas aeruginosa isolates, imipenem resistance was higher in the ICUs. For ciprofloxacin, resistance increased from 2.5% in 1991 to 13% in 1999 in the whole hospital, with similar figures for the ICUs. CONCLUSION: The resistance rates at the Karolinska Hospital were still generally low, but were increasing for some antibiotic-microbe combinations. The results emphasize the importance of including all sectors of a hospital in resistance surveillance studies, and also the value of long surveillance periods. PMID- 12848752 TI - Anaerobic bacteria commonly colonize the lower airways of intubated ICU patients. AB - OBJECTIVES: To investigate respiratory tract colonization by aerobic and anaerobic bacteria in mechanically ventilated patients. METHODS: Bacterial colonization of the stomach and the respiratory tract was qualitatively and quantitatively analyzed over time in 41 consecutive mechanically ventilated patients in a Swedish intensive care unit (ICU), with special emphasis on elucidation of the role of anaerobic bacteria in the lower respiratory tract. Samples were taken from the oropharynx, gastric juice, subglottic space and trachea within 24 h (median 14 h) of intubation, and then every third day until day 18 and every fifth day until day 33. RESULTS: The patients were often heavily colonized with microorganisms not considered to belong to a healthy normal oropharyngeal and gastric flora on admission to the ICU. A majority harbored enterococci, coagulase-negative staphylococci and Candida spp. in at least one site on day 1. Anaerobic bacteria, mainly peptostreptococci and Prevotella spp., were isolated from subglottic and/or tracheal secretions in 59% of the patients. Different routes of tracheal colonization for different groups of microorganisms were found. Primary or concomitant colonization of the oropharynx with staphylococci, enterococci, enterobacteria and Candida was often seen, while Pseudomonas spp., other non-fermenting Gram-negative rods and several anaerobic species often primarily colonized the trachea, indicating exogenous or direct gastrointestinal routes of colonization. CONCLUSIONS: Mechanically ventilated patients were heavily colonized in their lower airways by potential pathogenic microorganisms, including a high load of anaerobic bacteria. Different routes of colonization were shown for different species. PMID- 12848753 TI - Optimizing antibiotic therapy-the Aberdeen experience. AB - OBJECTIVE: To study the quality and continuity of treatment in the Acute Medicines Assessment Unit (AMAU) with regard to empirical prescription of antibiotics, mode of administration, adherence to ward antibiotic policy, as well as collection, awareness and utilization of microbiological investigations. METHODS: A prospective study over a 3-month period at the AMAU, Aberdeen Royal Infirmary (ARI), a teaching hospital in north-eastern Scotland, was performed. The study included all patients started on empirical antibiotics on admission to the AMAU and followed up until their discharge. RESULTS: Of 1303 patients admitted, 221 (17%) were started on empirical antibiotics. This was in accordance with hospital antibiotic policy in 52% of cases. Appropriate specimens were taken from 77% of patients. Culture results showed that 29% (n = 65) of the patients had clinically significant growth of organisms. Of the 65 patients with clinically significant culture results, 49% (n = 32) were on an inappropriate empirical regimen. In 55%, the medication was not changed to a more appropriate antibiotic. In 72% of the patients with a negative culture, the culture report had no obvious effect on the duration or type of antibiotic being administered. Intravenous antibiotics were used in 60% of patients. CONCLUSION: This study demonstrates a significant overuse of antibiotics, especially intravenous forms, despite a paucity of positive sepsis parameters and chest X-ray findings in these patients The duration of treatment could be shortened and an early switch policy introduced if culture results and sepsis profiles were taken into consideration, as there was a large number of unproven infections. Suggestions are made about how these improvements in prescribing could be made within the current administrative set-up of AMAUs. PMID- 12848754 TI - The influence of inadequate empirical antimicrobial treatment on patients with bloodstream infections in an intensive care unit. AB - OBJECTIVE: To determine the occurrence of inadequate antimicrobial therapy among critically ill patients with bacteremia and the factors associated with it, to identify the microorganisms that received inadequate antimicrobial treatment, and to determine the relationship between inadequate treatment and patients outcome. METHODS: From June 1995 to January 1999 we collected data on all clinically significant ICU-bacteremias in our teaching hospital. Clinical and microbiological characteristics were recorded and the adequacy of empirical antimicrobial treatment in each case was determined. We defined inappropriate empirical antimicrobial treatment as applying to infection that was not being effectively treated at the time the causative microorganism and its antibiotic susceptibility were known. Multivariate analysis was used to determine the variables associated with inappropriate empirical antimicrobial treatment and to evaluate the influence of this on the related mortality to bacteremia, using the SPSS package (9.0). RESULTS: Among 166 intensive care unit patients with bacteremia, 39 (23.5%) received inadequate antimicrobial treatment. In this last group the mean age of patients was 64.1 +/- 13.2 years, and 64% were men. Bacteremia was hospital-acquired in 92% of these cases. Eleven percent developed septic shock and 37.7% severe sepsis, and ultimately fatal underlying disease was present in 28.2% of patients given inadequate empirical antimicrobial treatment. The main sources of bacteremias in this group were: a vascular catheter (15.3%), respiratory (7.6%) or unknown (53.8%). The microorganisms most frequently isolated in the group with inadequate empirical antimicrobial treatment were: coagulase-negative staphylococci (29.5%), Acinetobacter baumannii (27.3%), Enterococcus faecalis, Pseudomonas aeruginosa, Enterobacter cloacae, Proteus mirabilis, Escherichia coli, and Candida species (4.5% each). The frequency of coagulase-negative staphylococci in the cases with inappropriate treatment was higher than in the group with appropriate treatment (OR 2.62; 95% CI: 1.10-6.21; P = 0.015). The global mortality rate was 56% and the related mortality was 30% in the group with inadequate empirical antimicrobial treatment. The only factor associated with inappropriate empirical antibiotic treatment was the absence of abdominal or respiratory focus (P = 0.04; OR = 0.35; 95% CI: 0.12-0.97). Septic shock was related to attributable mortality (P = 0.03; OR = 3.19; 95% CI: 1.08 9.40), but not inappropriate empirical antibiotic treatment (P = 0.24; OR = 1.71; 95% CI: 0.66-4.78). CONCLUSION: Almost a quarter of critically ill patients with bloodstream infections were given inadequate empirical antibiotic treatment, but mortality was not higher in the group with inadequate treatment than in the group with adequate treatment. This fact was probably due to microbiological factors and clinical features, such as the type of microorganism most frequently isolated and the source of the bacteremia. PMID- 12848755 TI - Clinical microbiological case: 'soap bubbles' in the cerebellum of an HIV infected patient. PMID- 12848756 TI - Clinical microbiological case: esophageal lesion in an AIDS patient. PMID- 12848757 TI - Human immunodeficiency virus type 1 recombinant B/G subtypes circulating in Coimbra, Portugal. AB - An increasing prevalence of HIV-1 non-B variants is being noticed in several European regions, particularly in countries such as Portugal, which have closer contacts with African endemic areas, where multiple HIV subtypes cocirculate. HIV 1 subtyping by phylogenetic analyses of reverse transcriptase, protease and env (C2-V3) genomic regions was carried out in plasma collected from 18 HIV-1 infected subjects living in Coimbra, Portugal, and suspected to be infected with non-B variants. Three (16.7%) subjects carried recombinant B/G viruses (BV3/BRT/Gpro; GV3/URT/Bpro; AV3/GRT/Bpro), whereas all the remaining individuals were infected with HIV-1 subtype B. This is the first report of recombinant B/G subtypes in Portugal. PMID- 12848758 TI - Neisseria elongata endocarditis: case report and literature review. AB - We describe a case of infective endocarditis due to Neisseria elongata, and review the literature. N. elongata is a constituent of the normal oral flora and a rare cause of infective endocarditis. Unfamiliarity with the organism and its rod-shaped morphology may lead to a delay in microbiological diagnosis. Although the organism is relatively sensitive to antibiotics, our experience in the management of the described case and a review of previous reports suggest that antibiotic therapy alone may not be sufficient. It is likely that patients with N. elongata endocarditis will require surgery. PMID- 12848759 TI - Antimicrobial susceptibility of 840 clinical isolates of Haemophilus influenzae collected in four European countries in 2000-2001. AB - In 2000-2001, 840 clinical isolates of Haemophilus influenzae were collected from laboratories in France, Germany, Italy and Spain (210 isolates/country). Beta Lactamase production among the isolates varied considerably by country, ranging from 8.1% in Germany to 34.8% in France. H. influenzae from patients or=18 years (16.5%). All isolates were susceptible to amoxicillin-clavulanate, ciprofloxacin and levofloxacin; 99.6% and 98.9% of isolates were susceptible to azithromycin and cefuroxime, respectively. Among the macrolides tested, azithromycin (MIC90, 2 mg/L) was eight-fold more potent than clarithromycin (MIC90, 16 mg/L) and roxithromycin (MIC90, 16 mg/L). Despite variations in beta-lactamase production between different countries, > 99% of all isolates were susceptible to amoxicillin-clavulanate, ciprofloxacin, levofloxacin, and azithromycin. PMID- 12848760 TI - A case of pleurisy associated with antibodies to Rickettsia conorii. AB - Rickettsia conorii is endemic in Mediterranean area. We describe an unusual sace of R. Conorii infection, which concerns a farmer with clinical, radiological and cytological findings of pleurisy without evidence of malignancy. An elevated antibody titre for R. Conorii was observed, using an indirect immunofluorescent antibody test. After treatment with Doxycycline, the patient presented a significant improvement of his clinical and radiological image and a four-fold decrease of the antibody titre for R. conorii. PMID- 12848761 TI - Cryptococcus neoformans var neoformans resistant to fluconazole in an HIV negative patient with chronic lymphocytic leukemia. AB - Cases of fluconazole-resistant Cryptococcus neoformans have been reported in AIDS patients previously treated with fluconazole. We report a case of fluconazole resistant cryptococcal meningitis in an HIV-negative patient not previously exposed to fluconazole. The patient experienced a clinical relapse after discontinuation of therapy with amphotericin B and subsequent initiation of fluconazole therapy. In vitro resistance was initially verified by Etest and tablet diffusion and later confirmed by NCCLS broth microdilution. PMID- 12848762 TI - Nocardia farcinica as the causative agent in a primary psoas abscess in a previously healthy cattle inspector. AB - A 42-year-old, previously healthy cattle inspector presented with a 7-day history of fever, a painful left knee, malaise and muscular pain. He did not suffer from an underlying disease, nor was he immunocompromised. After 12 days of hospitalization, a unilocular abscess in the left psoas muscle was diagnosed. Nocardia farcinica was isolated from the aspirate. No connection with his work could be demonstrated. The patient was successfully treated with trimethoprim sulfamethoxazole for 11 months. PMID- 12848763 TI - Evaluation of the WIDER I system for antimicrobial susceptibility testing of clinical isolates of Haemophilus influenzae and Streptococcus pneumoniae. AB - The aim of this study was to evaluate the WIDER I system for susceptibility testing of Haemophilus influenzae and Streptococcus pneumoniae. MICs of 12 antimicrobials against 42 H. influenzae and 58 S. pneumoniae strains were determined using 1W MIC panels and compared with those obtained by microdilution. Overall essential agreements were >99%. Very major errors were not detected. Major errors occurred with ampicillin (1.7% H. influenzae). Minor errors were 2.3% (amoxicillin-clavulanate, cefuroxime, chloramphenicol), 7.1% (ampicillin) and 16.7% (clarithromycin) for H. influenzae, and 1.7% (chloramphenicol, erythromycin, meropenem), 3.4% (amoxicillin-clavulanate, cefuroxime, tetracycline) and 8.6% (levofloxacin) for S. pneumoniae. The WIDER I system is a reliable method for susceptibility testing of H. influenzae and S. pneumoniae. PMID- 12848764 TI - Measurement of serum levels of eosinophil cationic protein in the diagnosis of acute gastrointestinal anisakiasis. AB - Thirty-two patients with abdominal pain and/or intestinal pseudo-obstruction who had consumed raw or undercooked fish in the previous 72 h, were included in a study of anisakiasis, a parasitation of the human gastrointestinal tract by third stage Anisakis simplex larvae. Skin prick test (SPT) against A. simplex were positive in all the patients. High median eosinophil cationic protein (ECP) serum concentrations (> 15 mg/L) at day 0 with normal serum levels at day 30 and a rise of median total and specific IgE against A. simplex at day 30, were observed. We conclude that a raised serum level of ECP in the first 72 h from the onset of symptoms coinciding with a positive SPT against A. simplex and high total and specific immunoglobulin (IgE) in the first month after the parasitation, could be a useful tool in the diagnosis of gastrointestinal anisakiasis, even if the parasite cannot be isolated. PMID- 12848765 TI - Some epidemiologic features of Campylobacter jejuni/coli infections in Bosnia and Herzegovina after the war. PMID- 12848769 TI - Angiotensin II dose-effect curves and Schild regression plots for characterization of different angiotensin II AT1 receptor antagonists in clinical pharmacology. AB - The 'Schild regression' method is based on the principle of assessing the rightward shift of agonist dose-effect curves in the presence of different doses/concentrations of the respective receptor antagonist and presenting their relationship in a double log plot (i.e. the 'Schild plot'). The original method was developed to quantitatively characterize antagonistic drugs in experimental pharmacology. The method was adopted for evaluation of various AT1 antagonists in humans utilizing (human) angiotensin II as the agonist. Angiotensin II (Ang II) in continuous intravenous dose-incremental administration resulted in a clearly dose-dependent increase in blood pressure. All AT1 antagonists tested after oral administration yielded concentration-dependent rightward shifts of those Ang II dose-effect curves that were quantified as dose ratio (DR). DR minus 1 (DR-1) enabled the assessment of antagonist time kinetics in humans and a quantitatively precise determination of the half-life of antagonism in vivo. Schild plots allowed for assessment of apparent Ki doses indicative of a twofold rightward shift of the Ang II effect, thus providing the means for a rational comparison of the pharmacological potency of many of these compounds, where the Ki doses obtained at 24 h after administration were in the range of 'therapeutic' doses. Schild plots of a variety of substances showed linear relations independent of whether the blockade was deemed surmountable or not. It is therefore assumed that this property does not play a role at clinical doses/concentrations. Slopes slightly below 1 in the Schild plots of all tested antagonists point to a second 'counterregulatory' vasodilatory mechanism of action of Ang II which becomes apparent with AT1 blockade in conditions of high doses/concentrations of Ang II. Concentration vs. effect relationships indicate that if assessed at the same degree of direct vascular antagonism, other effects, such as increase in plasma renin activity, may be present to a varying degree with different antagonists. Thus for irbesartan, the potency to stimulate renin release was found to be at least twice that of candesartan. These observations should stimulate further research into the relevance of these dynamic differences between the various compounds. Thus, methodologies relying on fundamental principles of experimental pharmacology can provide the clinical pharmacologist with powerful tools to measure accurately degree of antagonism and time kinetics and to investigate the nature of receptor antagonism in humans. PMID- 12848770 TI - Effects of fluticasone vs. fluticasone/salmeterol on airway calibre and airway hyperresponsiveness in mild persistent asthma. AB - AIMS: Inhaled corticosteroids alone or in combination with long acting beta2 agonists are indicated for use in mild persistent asthmatics. We set out to evaluate effects on airway hyperresponsiveness (AHR) and airway calibre using hydrofluoroalkane fluticasone/salmeterol (FP/SM) vs. double the dose of fluticasone alone (FP). METHODS: Fourteen mild persistent asthmatics completed a randomized double-blind crossover study with 1-week run-in and washout periods prior to treatments. Subjects received 3 weeks of FP 250 microg or FP 125 microg/SM 25 microg as 1 puff twice daily. Methacholine PD20 and lung function were measured after both baseline and treatment periods. RESULTS: There were no significant differences in baseline values prior to randomized treatments. Compared with pooled baseline, FP/SM and FP conferred improvements (P < 0.05) on methacholine PD20: 2.5 (95% confidence interval 1.7, 3.2) and 1.6 (0.8-2.3) doubling dose improvements, respectively; between FP/SM vs. FP there was a 0.9 (0.4, 1.4) doubling dose difference (P < 0.05). For forced expiratory volume in 1 s (FEV1), forced mid-expiratory flow (FEF25-75) and morning peak expiratory flow (PEF), FP/SM but not FP conferred improvements (P < 0.05) compared with pooled baseline, with FP/SM being greater than FP (P < 0.05): differences in FEV1 of 7.2% (3.8, 10.6) predicted, FEF25-75 of 11.2% (6.3, 16.1) predicted, and morning PEF of 17 L x min(-1)(1-32). CONCLUSIONS: FP/SM conferred improvements on AHR and airway calibre, while twice the dose of FP improved only AHR in patients with mild asthma. The differential effects of FP/SM and FP suggest separate but complementary actions of the two moieties on airway inflammation and smooth muscle stabilization. This may explain the beneficial effects of combination inhalers on exacerbations. PMID- 12848771 TI - Pseudoephedrine: effects on milk production in women and estimation of infant exposure via breastmilk. AB - AIMS: To assess the effects of pseudoephedrine on breast blood flow, temperature and milk production, and to estimate the likely infant dose during breastfeeding. METHODS: Eight lactating women (mean age 35 years and weight 69 kg) participated in a single-blind randomized crossover study of 60 mg pseudoephedrine hydrochloride vs placebo. Breast blood flow and surface temperature were measured from 0 to 4 h following the dose, and change in plasma prolactin was measured as the difference between predose and 1 h postdose concentrations. Milk production was measured for 24 h following placebo and pseudoephedrine. Infant dose of pseudoephedrine for a 60-mg dose administered four times daily to the mother was quantified as the product of average steady-state drug concentration in milk and an estimated milk production rate of 0.15 l x kg(-1) x day(-1) and expressed relative to the maternal weight-adjusted dose. RESULTS: There were no physiologically significant changes in breast blood flow or temperature between the placebo and pseudoephedrine periods. The mean change in plasma prolactin was slightly (13.5%), but not significantly lower (t = 1.245, P = 0.253) after pseudoephedrine (1775 mU x l(-1)) compared with placebo (2014 mU x l(-1)). However, the mean milk volume was reduced by 24% from 784 ml x day(-1) in the placebo period to 623 ml x day(-1) in the pseudoephedrine period (difference between means 161 ml x day(-1) (95% CI: 63, 259 ml x day(-1)); t = 3.9, P = 0.006). Assuming maternal intake of 60 mg pseudoephedrine hydrochloride four times daily, the estimated infant dose of pseudoephedrine was 4.3% (95% CI, 3.2, 5.4%) of the weight-adjusted maternal dose. CONCLUSIONS: A single dose of pseudoephedrine significantly reduced milk production. This effect was not attributable to changes in blood flow, but depression of prolactin secretion may be a contributing factor. At the maximum recommended pseudoephedrine doses, the calculated infant dose delivered via milk was < 10% of the maternal dose, and is unlikely to affect the infant adversely. The ability of pseudoephedrine to suppress lactation suggests a novel use for the drug. PMID- 12848772 TI - Population pharmacokinetics of intramuscular gentamicin administered to young infants with suspected severe sepsis in Kenya. AB - AIMS: To determine the population pharmacokinetics of intramuscular (i.m.) gentamicin in African infants with suspected severe sepsis. METHODS: Samples were withdrawn 1 h after a single i.m. injection of 8 mg x kg(-1) gentamicin and the next morning prior to any further dosing. Concentration-time data were analysed with the population pharmacokinetic package NONMEM. Data were fitted using a one compartment model with a log-normal model for interindividual variability and an additive residual error model. The influence of a range of clinical characteristics was tested on the pharmacokinetics of intramuscular gentamicin and the effect of incorporating interindividual variability on bioavailability was examined. RESULTS: The data set comprised 107 patients and 203 concentrations. Peak concentrations ranged from 3.0 mg x L(-1) to 19.8 mg x L(-1) (median 10.6 mg x L(-1)) and 'next day' samples from 0.3 mg x L(-1) to 6.2 mg x L(-1). The best models were clearance/bioavailability (CL) (L x h(-1)) = 0.0913 x weight (kg) x (age (days) + 1)/11)0.130 and volume of distribution/bioavailability (V) = 2.02 x (1 + 0.277 x (weight -3)). Therefore, an infant with the median weight of 3 kg and age 10 days would have a predicted CL of 0.274 L x h(-1) and V of 2.02 L. Interindividual variability in CL was 40% and in V was 42%. This model required a term for covariance between CL and V. When variability in bioavailability was introduced as an alternative model, interindividual variability in CL was 22%, in V 18% and in relative bioavailability 36%. CONCLUSIONS: Intramuscular administration of 8 mg x kg(-1) gentamicin daily to infants gives mean 1 h peak concentration of 10.6 mg x L(-1) and a trough concentration of less than 2 mg x L(-1). Wide variability in the peak concentration may reflect variable absorption rate or bioavailability. PMID- 12848773 TI - Contribution of increased oral bioavailability and reduced nonglomerular renal clearance of digoxin to the digoxin-clarithromycin interaction. AB - AIMS: A clinically important interaction between the cardiac glycoside digoxin and the antibiotic clarithromycin has been suggested in earlier reports. The aim of this study was to investigate the extent of the interaction and the relative contribution of different mechanisms. METHODS: In a randomized, placebo controlled, double-blind cross-over design single oral doses of 0.75 mg digoxin with oral coadministration of placebo or 250 mg clarithromycin twice daily for 3 days were administered to 12 healthy men. Additionally, three of the subjects received single intravenous doses of 0.01 mg x kg(-1) digoxin with oral placebo or clarithromycin. Digoxin plasma and urine concentrations were determined by a highly sensitive radioimmunoassay. RESULTS: Oral coadministration of clarithromycin resulted in a 1.7-fold increase of the area under the digoxin plasma concentration-time curve [mean AUC(0,24) +/- SD 23 +/- 5.2 vs. 14 +/- 2.9 microg x L(-1) x h; 95% confidence interval (CI) on the difference 7.0, 12; P = 0.002] and in a reduction of the nonglomerular renal clearance of digoxin [mean ClRng(0, 24) +/- SD 34 +/- 39 vs. 57 +/- 41 mL min-1; 95% CI on the difference 7.2, 45; P = 0.03]. The ratios of mean digoxin plasma concentrations with and without clarithromycin were highest during the absorption period of clarithromycin. After intravenous administration digoxin AUC(0,24) increased only 1.2-fold during coadministration of clarithromycin. CONCLUSIONS: Increased oral bioavailability and reduced nonglomerular renal clearance of digoxin both contribute to the interaction between digoxin and clarithromycin, probably due to inhibition of intestinal and renal P-glycoprotein. PMID- 12848774 TI - Lack of pharmacokinetic interaction between retigabine and phenobarbitone at steady-state in healthy subjects. AB - AIMS: To evaluate potential pharmacokinetic interactions between phenobarbitone and retigabine, a new antiepileptic drug. METHODS: Fifteen healthy men received 200 mg of retigabine on day 1. On days 4-32, phenobarbitone 90 mg was administered at 22.00 h. On days 26-32, increasing doses of retigabine were given to achieve a final dose of 200 mg every 8 h on day 32. The pharmacokinetics of retigabine were determined on days 1 and 32, and those for phenobarbitone on days 25 and 31. RESULTS: After administration of a single 200 mg dose, retigabine was rapidly absorbed and eliminated with a mean terminal half-life of 6.7 h, a mean AUC of 3936 ng x ml(-1) x h and a mean apparent clearance of 0.76 l x h(-1) x kg( 1). Similar exposure to the partially active acetylated metabolite (AWD21-360) of retigabine was observed. After administration of phenobarbitone dosed to steady state, the pharmacokinetics of retigabine at steady-state were similar (AUC of 4433 ng x ml(-1) x h and t1/2 of 8.5 h) to those of retigabine alone. The AUC of phenobarbitone was 298 mg x l(-1) x h when administered alone and 311 mg x ml(-1) x h after retigabine administration. The geometric mean ratios and 90% confidence intervals of the AUC were 1.11 (0.97, 1.28) for retigabine, 1.01 (0.88, 1.06) for AWD21-360 and 1.04 (0.96, 1.11) for phenobarbitone. Individual and combined treatments were generally well tolerated. One subject was withdrawn from the study on day 10 due to severe abdominal pain. Headache was the most commonly reported adverse event. No clinically relevant changes were observed in the electrocardiograms, vital signs or laboratory measurements. CONCLUSIONS: There was no pharmacokinetic interaction between retigabine and phenobarbitone in healthy subjects. No dosage adjustment is likely to be necessary when retigabine and phenobarbitone are coadministered to patients. PMID- 12848775 TI - A population pharmacokinetic model for paclitaxel in the presence of a novel P-gp modulator, Zosuquidar Trihydrochloride (LY335979). AB - AIMS: To develop a population pharmacokinetic model for paclitaxel in the presence of a MDR modulator, zosuquidar 3HCl. METHODS: The population approach was used (implemented with NONMEM) to analyse paclitaxel pharmacokinetic data from 43 patients who received a 3-h intravenous infusion of paclitaxel (175 mg x m(-2) or 225 mg x m(-2)) alone in cycle 2 or concomitantly with the oral administration of zosuquidar 3HCl in cycle 1. RESULTS: The structural pharmacokinetic model for paclitaxel, accounting for the Cremophor ELTM impact, was a three-compartment model with a nonlinear model for paclitaxel plasma clearance (CL), involving a linear decrease in this parameter during the infusion and a sigmoidal increase with time after the infusion. The final model described the effect of Zosuquidar 3HCl on paclitaxel CL by a categorical relationship. A 25% decrease in paclitaxel CL was observed, corresponding to an 1.3-fold increase in paclitaxel AUC (from 14829 microg x l(-1) x h to 19115 microg x l(-1) x h following paclitaxel 175 mg x m(-2)) when zosuquidar Cmax was greater than 350 microg x l(-1). This cut-off concentration closely corresponded to the IC50 of a sigmoidal Emax relationship (328 microg x l(-1)). A standard dose of 175 mg x m( 2) of paclitaxel could be safely combined with doses of zosuquidar 3HCl resulting in plasma concentrations known, from previous studies, to result in maximal P-gp inhibition. CONCLUSIONS: This analysis provides a model which accurately characterized the increase in paclitaxel exposure, which is most likely to be due to P-gp inhibition in the bile canaliculi, in the presence of zosuquidar 3HCl (Cmax > 350 microg x l(-1)) and is predictive of paclitaxel pharmacokinetics following a 3 h infusion. Hence the model could be useful in guiding therapy for paclitaxel alone and also for paclitaxel administered concomitantly with a P-gp inhibitor, and in designing further clinical trials. PMID- 12848776 TI - Physiologically based modelling of inhibition of metabolism and assessment of the relative potency of drug and metabolite: dextromethorphan vs. dextrorphan using quinidine inhibition. AB - AIMS: To define the relative antitussive effect of dextromethorphan (DEX) and its primary metabolite dextrorphan (DOR) after administration of DEX. METHODS: Data were analysed from a double-blind, randomized cross-over study in which 22 subjects received the following oral treatments: (i) placebo; (ii) 30 mg DEX hydro-bromide; (iii) 60 mg DEX hydro-bromide; and (iv) 30 mg DEX hydro-bromide preceded at 1 h by quinidine HCl (50 mg). Cough was elicited using citric acid challenge. Pharmacokinetic data from all non-placebo arms of the study were fitted simultaneously. The parameters were then used as covariates in a link PK PD model of cough suppression using data from all treatment arms. RESULTS: The best-fit PK model assumed two- and one-compartment PK models for DEX and DOR, respectively, and competitive inhibition of DEX metabolism by quinidine. The intrinsic clearance of DEX estimated from the model ranged from 59 to 1536 l x h( 1), which overlapped with that extrapolated from in vitro data (12-261 l x h(-1)) and showed similar variation (26- vs. 21-fold, respectively). The inhibitory effect of quinidine ([I]/Ki) was 19 (95% confidence interval of mean: 18-20) with an estimated average Ki of 0.017 microM. Although DEX and DOR were both active, the potency of the antitussive effect of DOR was 38% that of DEX. A sustained antitussive effect was related to slow removal of DEX/DOR from the effect site (ke0 = 0.07 h(-1)). CONCLUSIONS: Physiologically based PK modelling with perturbation of metabolism using an inhibitor allowed evaluation of the antitussive potency of DOR without the need for separate administration of DOR. PMID- 12848777 TI - CYP2D6, GST-M1 and GST-T1 enzymes: expression in parathyroid gland and association with the parathyroid hormone concentration during early renal replacement therapy. AB - AIMS: The purpose of this research was to characterize CYP2D6, GST-M1 and GST-T1 enzyme expression in human parathyroid tissue, and to determine whether or not there is any association between deficiencies in these enzymes and serum parathyroid hormone concentrations in patients with end-stage renal disease. METHODS: Surgical human parathyroid tissue was obtained and evaluated by immunohistochemistry for cellular localization of CYP2D6, GST-M1 and GST-T1 and colocalization of CYP2D6 with parathyroid hormone. Blood samples were collected from 328 Caucasian patients with end-stage renal disease for genetic testing of CYP2D6*3, *4, *5, *6, *7 and GST-M1*0 and GST-T1*0 alleles. Clinical chemistry data and serum intact parathyroid hormone (iPTH) concentrations were obtained from patient medical records. In 277 of the patients, the same laboratory performed all clinical tests. RESULTS: CYP2D6, GST-M1 and GST-T1 were present in human parathyroid tissue. CYP2D6 was colocalized with parathyroid hormone in parathyroid chief cells. Within the end-stage renal disease population, a nonfunctional CYP2D6 genotype was present in 18.2%[95% confidence interval (CI) 8.0, 28.4] of patients in the 1st iPTH concentration quintile (iPTH < 64 pg x mL( 1)), in 0% (95% CI 0, 7.5) of those in the 2nd quintile, in 1.8% (95% CI 0, 9.3) of those in the 3rd quintile, in 9.1% (95% CI 1.5, 16.7) of those in the 4th quintile, and in 16.7% (95% CI 6.8, 26.5) of those in the 5th quintile (iPTH > 347 pg x mL(-1)) (P = 0.001). Out of 12 CYP2D6-deficient females, seven were in the 1st iPTH concentration quintile and the remaining five were in the 5th quintile. Patients deficient in the GST-M1 and GST-T1 enzymes displayed a far more uniform frequency distribution relative to serum iPTH concentrations. CONCLUSIONS: The presence of CYP2D6, GST-M1 and GST-T1 in parathyroid cells was observed. An association is reported between a lack of CYP2D6 and iPTH concentrations in newly diagnosed end-stage renal disease patients. Gender and concomitant deficiency in GST-M1 and/or GST-T1 appear to define this association further. It remains to be established whether these associations reflect a cause effect relationship between deficient expression of metabolizing enzymes and severity of secondary manifestation of renal failure. PMID- 12848778 TI - Frequency of C3435T single nucleotide MDR1 genetic polymorphism in an Asian population: phenotypic-genotypic correlates. AB - AIMS: To investigate the frequency of the single nucleotide polymorphism C3435T in exon 26 of the MDR1 gene in Asians and to determine the functional significance of this SNP with the clinical pharmacokinetics of oral cyclosporin (Neoral) in 10 stable heart transplant patients. METHODS: The MDR1 C3435T polymorphism was investigated in 290 healthy Asian subjects (98 Chinese, 99 Malays and 93 Indians). We also compared the MDR1 polymorphism between the Asian population studied here and the published data on Africans and Caucasians. The clinical relevance of this SNP on oral bioavailability of a known P-gp substrate, cyclosporin, was assessed in 10 stable Chinese heart transplant patients. RESULTS: The homozygous TT genotype was observed in 32%, 28% and 43% of Chinese, Malays and Indians. The homozygous CC genotype was found in 25% of Chinese and Malays compared with 18% of Indians. The Indians had a lower frequency of the C allele [0.38 (0.31-0.45)] compared with the Chinese [0.46 (0.39-0.53)] and Malays [0.48 (0.42-0.55)]. Chi-squared test showed that the distribution of allele frequencies between the Malays and Indians differed significantly (P = 0.04). In this Asian population, the overall distribution of genotypes (CC, CT and TT) and allele frequencies were significantly different from those in Africans (P < 0.001). The results were also significant when the Chinese, Malays and Indians were compared separately with the African group (P < 0.001). Compared with the Caucasian data, the overall distribution of genotype and allele frequencies in the Asian population were also significantly different (P < or = 0.05). However, when each Asian ethnic group was compared separately with the Caucasians, only the Indians were found to be significantly different (P < or = 0.004). Genotypic phenotypic correlations of this SNP were assessed in 10 stable Chinese heart transplant patients. The median AUC(0,4 h) was 11% lower in patients with CC genotype compared with subjects with TT genotype. However, the interpatient variability in AUC(0,4 h) was high in patients, especially in those with CC genotype. CONCLUSIONS: The distribution of the SNP C3435T in exon 26 in the Chinese and Malay population was found to be similar to the Caucasians whereas the Indians were different. The Asian population also differed significantly from the African and Caucasian population in the distribution of the C3435T SNP. The low frequency of the T allele in the Indian population implies lower expression of P-gp and may have important therapeutic and prognostic implications for use of P-gp dependent drugs in individuals of Indian origin. PMID- 12848779 TI - Discontinuation and switching of therapy after initiation of lipid-lowering drugs: the effects of comorbidities and patient characteristics. AB - AIMS: To evaluate the effects of comorbidities and patient characteristics on treatment continuation among patients starting their first course of lipid lowering drug (LLD) therapy. METHODS: Within the UK General Practice Research Database (GPRD), we identified 22 408 patients who started LLD therapy due to coronary heart disease, hyperlipidaemia, or other atherosclerotic diseases, and who received > or = two prescriptions for LLD between January 1 1990 and December 31 1997. Differences in potential predictors of treatment continuation between patients who continued, and patients who discontinued/switched lipid-lowering therapy within 1 year after treatment initiation were compared by fitting multivariate logistic regression models. The effects of baseline characteristics on treatment continuation after switching of LLDs were also analysed. RESULTS: Discontinuation/switching of lipid-lowering therapy was common during the study period, especially among patients who received nonstatin, nonfibrate LLDs (log rank test P = 0.0001). Statin use, more frequent physician visits, more concurrent cardiovascular medications, diabetes, and fewer noncardiovascular medications were associated with treatment continuation of LLDs. Among patients who switched therapy, prescribing of a statin as the substituted LLD, more concurrent cardiovascular medications, and later treatment switching were related to a higher probability of treatment continuation after switching LLDs. CONCLUSIONS: Treatment continuation after initiation or switching of lipid lowering therapy largely increased with concomitant cardiovascular comorbidities, and more health care utilization, and is more common for statins than for other LLDs. Practice guidelines, patient education, and quality of care assessment for lipid-lowering therapy should emphasize factors that predispose patients to discontinuation/switching, in an effort to optimize the choice of therapeutic regimens and to improve patient adherence. PMID- 12848780 TI - Antibiotic prescribing for children. Too much and too little? Retrospective observational study in primary care. AB - AIMS: To investigate the extent of dose-related off-label antibiotic paediatric prescribing in primary care and to identify any potential clinical effects, particularly of lower than recommended dose prescribing. METHODS: Assessment of antibiotic prescribing in 168 396 children aged 0-16 years for the year 1999-2000 from data retrieved from 158 general practices using the national Scottish primary care computer system GPASS. The setting was general practices in Scotland. RESULTS: During the study period at least one course of antibiotics was prescribed to 23 911 children (14.2%). A total of 4582 (19.2%) children were prescribed an antibiotic dose of less than that recommended in the Summary of Product Characteristics (SPC). The number of children prescribed an antibiotic at less than recommended dose increased with age from 1154 (11.8%) aged 0-4 years to 1827 (30.0%) in the 12-16 years age group. For each antibiotic, prescribing lower than recommended dose occurred most frequently at those ages at which a dose increase was recommended in the SPC. Antibiotic prescribing at less than the recommended dose was not associated with an increased return rate for further antibiotic prescriptions during the following month, but in 5-11-year-olds was associated with a significant 48% increase in the total number of antibiotic courses prescribed during the study year [mean = 2.09 [95% confidence interval (CI) 1.79, 2.39]vs. 1.41 [95% CI 1.35, 1.47]]. Antibiotic prescribing at doses higher than recommended occurred less frequently (1.6%) and decreased steadily with age. CONCLUSIONS: Off-label prescribing of antibiotics at less than the recommended dose in children is common in primary care and occurs primarily as the result of a failure to increase antibiotic dosage with age in line with SPC recommendations. Adoption of a uniform approach to SPC age banding for antibiotic dose increments would reduce the frequency of dose-related off-label antibiotic prescribing in children and help minimize the potential for the development of antibiotic resistance. PMID- 12848781 TI - Development of a dosing strategy for enoxaparin in obese patients. AB - AIMS: Enoxaparin dosing is currently based on total body weight. It is not known how to dose adjust for patients who are overweight or obese. This population pharmacokinetic pharmacodynamic (PKPD) study was undertaken to determine a suitable dosing strategy for such patients. METHODS: Patients admitted to the Royal Brisbane Hospital and prescribed enoxaparin as part of their normal care were eligible for inclusion into the study. Approximately three blood samples were taken per patient to determine anti-Xa concentrations. The occurrence of bruising was also recorded. A population pharmacokinetic-pharmacodynamic analysis using NONMEM was undertaken. Simulations were performed using MATLAB. RESULTS: Ninety-six patients were recruited in a prospective study. One-third of patients had a body mass index < 24.9 kg x m(-2), one-third from 25 to 29.9 kg x m(-2), and one-third> 30 kg x m(-2). A two-compartment linear model with additive error was fitted to the data. A covariate analysis showed clearance was best described by lean body weight and the central volume compartment by total body weight. The probability of bruising using a logistic regression model was best described by Cmax and age. Simulations suggest that patients over 50 years of age whose total body weight is> 90 kg, or under 50 years of age whose total body weight is> 120 kg are likely to have a smoother concentration-time profile and less bruising if a dose of 100 IU x kg(-1) (1 mg x kg(-1)) based on lean body weight is administered every 8 h. CONCLUSIONS: Dose adjustments of enoxaparin in obese patients are likely to reduce the prevalence of bruising, although prospective validation of this is required. PMID- 12848782 TI - Add-on therapy with montelukast or formoterol in patients with the glycine-16 beta2-receptor genotype. AB - AIMS: We assessed whether montelukast or formoterol provides additive effects to asthmatics not controlled on inhaled corticosteroids, by studying patients who were considered to be genetically susceptible to beta2-receptor down regulation and subsensitivity, and who expressed the homozygous glycine-16 beta2-receptor genotype. METHODS: Fifteen corticosteroid-treated, mild to moderate persistent asthmatics received montelukast 10 mg once daily or formoterol 9 micro g twice daily for 2 weeks, separated by a 2-week placebo run-in and washout, in a double blind, double-dummy, randomized crossover design. Bronchoprotection against adenosine monophosphate (AMP) challenge (primary endpoint), spirometry and blood eosinophils were measured at trough after placebo, first and last doses. RESULTS: For AMP PC20vs placebo, there were sustained significant (P < 0.05) doubling dilution improvements following first (1.1; 95% CI 0.4, 1.9) and last (1.0; 95% CI 0.3, 1.8) doses of montelukast, and following first (1.3; 95% CI 0.1, 2.6) but not last (0.3; 95% CI -0.9, 1.6) doses of formoterol. Blood eosinophils (x 10(6) l(-1)) were significantly (P < 0.05) suppressed after the last dose of montelukast (-71; 95% CI -3, -140) compared with placebo, while formoterol exhibited a nonsignificant rise (20; 95% CI -92, 132). Neither treatment significantly improved FEV1, FEF25-75 or PEF after 2 weeks. CONCLUSIONS: In genetically susceptible patients with the homozygous glycine-16 genotype, montelukast, but not formoterol, conferred sustained anti-inflammatory properties in addition to inhaled corticosteroid, which were dissociated from changes in lung function after 2 weeks. Thus, assessing lung function may miss potentially beneficial anti-inflammatory effects of montelukast when used as add-on therapy. PMID- 12848783 TI - Pharmacokinetics and clinical effects of phenytoin and fosphenytoin in children with severe malaria and status epilepticus. AB - AIMS: Status epilepticus is common in children with severe falciparum malaria and is associated with poor outcome. Phenytoin is often used to control status epilepticus, but its water-soluble prodrug, fosphenytoin, may be more useful as it is easier to administer. We studied the pharmacokinetics and clinical effects of phenytoin and fosphenytoin sodium in children with severe falciparum malaria and status epilepticus. METHODS: Children received intravenous (i.v.) phenytoin as a 18 mg kg-1 loading dose infused over 20 min followed by a 2.5 mg x kg(-1) 12 hourly maintenance dose infused over 5 min (n = 11), or i.v. fosphenytoin, administered at a rate of 50 mg x min(-1) phenytoin sodium equivalents (PE; n = 16), or intramuscular (i.m.) fosphenytoin as a 18 mg x kg(-1) loading dose followed by 2.5 mg x kg(-1) 12 hourly of PE (n = 11). Concentrations of phenytoin in plasma and cerebrospinal fluid (CSF), frequency of seizures, cardiovascular effects (respiratory rate, blood pressure, trancutaneous oxygen tension and level of consciousness) and middle cerebral artery (MCA) blood flow velocity were monitored. RESULTS: After all routes of administration, a plasma unbound phenytoin concentration of more than 1 microg x ml(-1) was rapidly (within 5-20 min) attained. Mean (95% confidence interval) steady state free phenytoin concentrations were 2.1 (1.7, 2.4; i.v. phenytoin, n = 6), 1.5 (0.96, 2.1; i.v. fosphenytoin, n = 11) and 1.4 (0.5, 2.4; i.m. fosphenytoin, n = 6), and were not statistically different for the three routes of administration. Median times (range) to peak plasma phenytoin concentrations following the loading dose were 0.08 (0.08-0.17), 0.37 (0.33-0.67) and 0.38 (0.17-2.0) h for i.v. fosphenytoin, i.v. phenytoin and i.m. fosphenytoin, respectively. CSF: plasma phenytoin concentration ratio ranged from 0.12 to 0.53 (median = 0.28, n = 16). Status epilepticus was controlled in only 36% (4/11) following i.v. phenytoin, 44% (7/16), following i.v. fosphenytoin and 64% (7/11) following i.m. fosphenytoin administration, respectively. Cardiovascular parameters and MCA blood flow were not affected by phenytoin administration. CONCLUSIONS: Phenytoin and fosphenytoin administration at the currently recommended doses achieve plasma unbound phenytoin concentrations within the therapeutic range with few cardiovascular effects. Administration of fosphenytoin i.v. or i.m. offers a practical and convenient alternative to i.v. phenytoin. However, the inadequate control of status epilepticus despite rapid achievement of therapeutic unbound phenytoin concentrations warrants further investigation. PMID- 12848784 TI - The human hepatic metabolism of simvastatin hydroxy acid is mediated primarily by CYP3A, and not CYP2D6. AB - AIMS: To identify the cytochrome P450 (CYP) isoforms responsible for the metabolism of simvastatin hydroxy acid (SVA), the most potent metabolite of simvastatin (SV). METHODS: The metabolism of SVA was characterized in vitro using human liver microsomes and recombinant CYPs. The effects of selective chemical inhibitors and CYP antibodies on SVA metabolism were assessed in human liver microsomes. RESULTS: In human liver microsomes, SVA underwent oxidative metabolism to three major oxidative products, with values for Km and Vmax ranging from about 50 to 80 microM and 0.6 to 1.9 nmol x min(-1) x mg(-1) protein, respectively. Recombinant CYP3A4, CYP3A5 and CYP2C8 all catalysed the formation of the three SVA metabolites, but CYP3A4 was the most active. CYP2D6 as well as CYP2C19, CYP2C9, CYP2A6, CYP1A2 did not metabolize SVA. Whereas inhibitors that are selective for CYP2D6, CYP2C9 or CYP1A2 did not significantly inhibit the oxidative metabolism of SVA, the CYP3A4/5 inhibitor troleandomycin markedly (about 90%) inhibited SVA metabolism. Quercetin, a known inhibitor of CYP2C8, inhibited the microsomal formation of SVA metabolites by about 25-30%. Immunoinhibition studies revealed 80-95% inhibition by anti-CYP3A antibody, less than 20% inhibition by anti-CYP2C19 antibody, which cross-reacted with CYP2C8 and CYP2C9, and no inhibition by anti-CYP2D6 antibody. CONCLUSIONS: The metabolism of SVA in human liver microsomes is catalysed primarily (> or = 80%) by CYP3A4/5, with a minor contribution (< or = 20%) from CYP2C8. CYP2D6 and other major CYP isoforms are not involved in the hepatic metabolism of SVA. PMID- 12848785 TI - Pharmacokinetics of cathinone, cathine and norephedrine after the chewing of khat leaves. AB - AIM: The stimulating herbal drug khat is habitually used in East Africa and the Arabian peninsula but is also imported into other countries. The aim was to study the pharmacokinetics of its alkaloids cathinone, cathine and norephedrine. METHODS: Four volunteers chewed khat leaves in an amount equivalent to one quarter of that used in a typical khat session. Blood samples were collected up to 80 h and the alkaloids were assayed using gas chromatography-mass spectrometry. The data were evaluated using computerized pharmacokinetic compartmental analysis. RESULTS: The plasma concentration-time data for the alkaloids could be described using a two-compartment model with two-segment absorption. The mucosa of the oral cavity is considered to be the first absorption segment, where the major proportion of the alkaloids is absorbed (mean +/- SD 59 +/- 21% for cathinone and 84 +/- 6% for cathine). The extraction of the alkaloids from the leaves by chewing was very effective with only 9.1 +/- 4.2% remaining as a residue. Cathinone was eliminated from the central compartment with a mean half-life of 1.5 +/- 0.8 h. The half-life of cathine was 5.2 +/- 3.4 h. The metabolism of cathinone to norephedrine had a substantial influence on its plasma concentration profile. Psychophysical functions were essentially unaffected by the chewing of khat. CONCLUSIONS: The pharmacokinetics of khat alkaloids in humans explain why chewing is the preferred form of khat ingestion. Subjects absorbed a mean dose of 45 mg of cathinone, and did not suffer any severe adverse reactions. PMID- 12848786 TI - Single-dose pharmacokinetics and tolerability of pegylated interferon-alpha2b in young and elderly healthy subjects. AB - AIMS: To assess the single-dose pharmacokinetics and tolerability of pegylated interferon-alpha2b (PEG-Intron) in young and elderly healthy subjects. METHODS: In this parallel-design study, a single 1 microg x kg(-1) PEG-Intron dose was given subcutaneously to 24 subjects in the age groups 20-45, 65-69, 70-74 and 75 80 years (n = 6/group). Blood sampling and tolerability assessments were performed up to 168 h postdose. RESULTS: The pharmacokinetic parameters were similar in all age groups. The elderly to young subject ratios for Cmax were 91.1, 79.5, and 107% for the 65-69 years, 70-74 years and 75-80 years groups, respectively. The corresponding values for AUC(0- infinity ) and CL/F were 111, 102 and 108%, and 82.5, 95.8 and 86.4%, respectively. Mean differences from the 20 to 45 years group and the 65-69 years, 70-74 years and 75-80 years groups for PEG-Intron Vd/F were 108, 128 and 104%, respectively. None of these differences was statistically significant based on ANOVA. Results from a Dunnett's test (as post hoc assessment) confirmed that the pharmacokinetic parameters of Group II, Group III or Group IV were not different from those of Group I. Almost all (23/24; 96%) subjects reported typical interferon-alpha side-effects (flu-like symptoms, headache). One elderly patient had a myocardial infarction 12 h postdose, but recovered fully. CONCLUSIONS: There are no pharmacokinetic reasons for initial dose adjustment of PEG-Intron based on age. PMID- 12848787 TI - Comparability of whole-blood and plasma clozapine and norclozapine concentrations. PMID- 12848788 TI - It is time to establish a subspecialty in noninvasive electrocardiology. PMID- 12848789 TI - Effects of P-wave dispersion on atrial fibrillation in patients with acute anterior wall myocardial infarction. AB - BACKGROUND: P-wave dispersion (P dispersion), defined as the difference between the maximum and the minimum P-wave duration (P minimum), and maximum P-wave duration (P maximum) have been used to evaluate the discontinuous propagation of sinus impulse and the prolongation of atrial conduction time respectively. The aim of this study was to investigate whether early assessment of P dispersion predicts paroxysmal atrial fibrillation (AF) in patients with acute anterior wall myocardial infarction (MI). METHODS: We prospectively evaluated 147 consecutive patients (45 women, 102 men; aged 55 +/- 9 years) with a first acute anterior wall MI. All patients were evaluated by echocardiography to measure the left atrial diameter and left ventricular ejection fraction (LVEF). Electrocardiography was recorded from all patients on admission and every day during hospitalization. RESULTS: AF occurred in 25 patients. In 122 patients, AF did not occur. P maximum was found to be significantly higher in patients with AF than in patients without AF (115 +/- 17.3 ms vs 101 +/- 14.7 ms, P = 0.001). P dispersion also was significantly higher in patients with AF than in patients without AF (50 +/- 12.5 ms vs 43 +/- 10.1 ms, P = 0.01). There was no significant difference between the two groups in P minimum (64 +/- 12.5 ms vs 59 +/- 11.7 ms, P = 0.057). The echocardiographically left atrial diameters were not significantly higher in the patients with AF than those without (25 +/- 3.38 mm and 23 +/- 3.36 mm, respectively, P = 0.76). LVEF was found to be significantly different in the patients who developed AF and in those who did not (37.96 +/- 6.18% vs 47.70 +/- 6.01%, P = 0.0001). CONCLUSIONS: Although P maximum and P dispersion are significant predictive factors of AF in patients with acute anterior wall MI in the univariate analysis, on the basis of multivariate analysis, only age and LVEF were independent predictive parameters for AF. PMID- 12848790 TI - Effects of slow coronary artery flow on QT interval duration and dispersion. AB - BACKGROUND: The coronary slow-flow phenomenon is an angiographic phenomenon characterized by delayed opacification of vessels in the absence of any evidence of obstructive epicardial coronary disease. Several studies have demonstrated myocardial ischemia in patients with slow coronary artery flow. In the present study, we aimed at evaluating the effects of slow coronary artery flow on QT interval duration and QT dispersion as a possible indicator of increased risk for ventricular arrhythmias and sudden cardiac death. METHODS: The study population included 49 patients with angiographically proven normal coronary arteries and slow coronary flow in all three coronary vessels (group I, 33 males, 16 females, mean age = 48 +/- 9 years), and 71 patients with angiographically proven normal coronary arteries without associated slow coronary flow (group II, 47 males, 24 females, mean age = 50 +/- 8 years). Coronary flow rates of all subjects were documented by thrombolysis in myocardial infarction frame count (TIMI frame count). QT interval duration and QT dispersion of all subjects were measured on the standard 12-lead electrocardiogram. RESULTS: There was no statistically significant difference between the two groups in respect to age, gender, presence of hypertension, and diabetes mellitus. There was a significant difference between the two groups in respect to the presence of cigarette smoking, typical angina, and positive exercise test results. TIMI frame counts of group I patients were significantly higher than those of group II patients for all three coronary arteries (P < 0.001). Maximum corrected QT interval (QTcmax) of group I did not differ from the QTcmax of group II (P > 0.05). However, minimum corrected QT interval (QTcmin) of group I was significantly lower than that for group II (P = 0.008). Consequently, corrected QT dispersion (QTcd) in group I was found to be significantly higher than in group II (P < 0.001). CONCLUSION: QTcd, indicating increased risk for ventricular arrhythmias and cardiovascular mortality, was found to be significantly higher in patients with slow coronary artery flow. However, further long-term prospective studies should be carried out to establish the significance of QTcd as a risk factor for ventricular arrhythmias and subsequent sudden cardiac death in patients with slow coronary artery flow. PMID- 12848791 TI - Utility of SAECG in arrhythmogenic right ventricle dysplasia. AB - BACKGROUND: Arrhythmogenic right ventricular dysplasia (ARVD) is characterized by progressive replacement of RV myocardium with fibro-adipose tissue thought to be responsible for the presence of late potentials (LP) detected by SAECG. The general consensus on the role of SAECG in the diagnosis and prognosis of patients with ARVD is lacking. The purpose of this systematic review was to better define the role of SAECG in ARVD. METHODS: An extensive review of literature was done to specifically describe the prevalence of LP in ARVD and its determinants, explore the various options available to improve the diagnostic ability of SAECG, and provide recommendations for proper utilization of this technique. RESULTS: LPs are frequent in ARVD (47-100%), and more prevalent in severe disease and in patients with documented spontaneous VT. SAECG is a useful test in following the characteristic evolutivity of the disease. 4-16% of normal family members of patients with ARVD also have abnormal SAECG results. Detection of LP in ARVD can be improved by employing a high-pass filter of 25 Hz and specifically looking for changes in the Z leads. CONCLUSIONS: SAECG testing should be considered a standard part of the evaluation of patients with known or suspected ARVD. Further research is needed to confirm the value of SAECG testing in predicting arrhythmia risk and assessing the rate of disease progression, as well as to determine if greater prevalence of SAECG abnormalities in family members of patients with ARVD represents early detection of ARVD. The ongoing multidisciplinary study of right ventricular dysplasia will hopefully answer some of these questions. PMID- 12848792 TI - Higher energy monophasic DC cardioversion for persistent atrial fibrillation: is it time to start at 360 joules? AB - BACKGROUND: Electrical direct-current cardioversion (DCCV) has become a routine therapy for atrial fibrillation (AF), although some uncertainty remains regarding the optimal energy settings. AIMS: This study examines whether the use of a higher initial energy monophasic shock of 360 joules (J) for external DCCV, in patients with persistent AF would prove more effective, yet as safe, as the use of a lower initial energy 200 J shock. METHODS: A cohort of 107 patients with persistent AF was prospectively randomized to an initial synchronized DCCV shock of 360 J versus 200 J (n = 50 vs 57), followed by a similar shock sequence thereafter of four further shocks of 360 J for the two groups. In all patients the levels of troponin I (cTnI) were measured precardioversion and 18-20 hours later, the following day. In a subgroup of 36 patients in each group, the levels of creatine kinase (CK) and aspartate transaminase (AST) were measured pre- and 18-20 hours postcardioversion. RESULTS: The success rate for DCCV was significantly higher in the 360 J group compared to the 200 J group (96.0% vs 75.4%, P = 0.003). The mean CK IU/L levels (1137.5.0 vs 2411.8, P = 0.014) and AST levels (39.83 vs 52.86, P = 0.010) were significantly lower in the 360 J group compared to the 200 J group. There was no statistical rise in cTnI (microg/L) in either group (P = 1.00). The average number of shocks delivered (1.84 vs 2.56, P = 0.006) was significantly less in the 360 J group than in the 200 J group, although total energy requirements for DCCV were similar for the two groups (662.4 J vs 762.4 J, P = 0.67). CONCLUSION: For patients with persistent AF the use of a higher initial-energy monophasic shock of 360 J achieves a significantly greater success rate, with less skeletal muscle damage (and no cardiac muscle damage) as compared with the traditional starting energy of a 200 J DC shock. PMID- 12848793 TI - Heart rate turbulence following ventricular premature beats in healthy controls. AB - BACKGROUND: Heart rate turbulence (HRT) has recently been described as a strong, independent risk stratifier in postinfarct patients. To date, however, the incidence of false positive HRT findings in adults is unknown. Therefore, we performed a blinded, retrospective analysis of HRT in a prospectively collected database of 110 apparently healthy persons to determine the prevalence and clinical significance of abnormal HRT findings in healthy controls using previously published cut-off values. METHODS AND RESULTS: The study included 43 out of 110 apparently healthy adults, in whom a sufficient number of ventricular premature beats were available for HRT analysis on 24-hour Holter recordings. The HRT slope was dichotomized at 2.5 ms per R-R interval and HRT onset was dichotomized at 0% using previously established criteria to define an abnormal HRT analysis in postinfarct patients. Using these definitions, abnormal HRT results were found in 2 out of 43 controls (5%) for HRT slope and in 8 out of 43 controls (19%) for HRT onset without any cardiac deaths during 32 +/- 15 months follow-up. CONCLUSIONS: The incidence of false positive HRT results in healthy middle-aged volunteers is low for HRT slope (5%), but not for HRT onset (19%) when previously published cut-off values are used to define abnormal HRT results. Thus, HRT slope dichotomized at 2.5 ms per R-R interval, but not HRT onset dichotomized at 0%, may be used as a relatively specific tool for risk stratification in middle-aged persons. PMID- 12848794 TI - Severity in myocardial dysfunction contributed to long-term fluctuation of heart rate, rather than short-term fluctuations. AB - BACKGROUND: It is known that heart rate shows complex behavior, long-term fluctuation of heart rate, and short-term fluctuations in heart failure. Analyzing these properties and examining the relationship to the disease, severity may increase the understanding of the background of heart rate variability (HRV). METHODS: In 61 patients (mean age 65 +/- 9 years, 32 ischemic heart disease, 29 cardiomyopathy), with myocardial dysfunction, 24-hour ambulatory electrocardiography was performed. After the construction of the time series of R-R intervals, 15 HRV parameters were measured, including mean heart rate, standard deviation of N-N intervals (SDNN), ratio of low frequency/high frequency power (LF/HF), HRV triangular index (TI), and ratio of length/width at the 90% level of all scattered points. RESULTS: By using the multiple regression analysis, we tested which HRV parameter (HR, SDNN, LF/HF, TI, or length/width) independently correlated with left ventricular ejection fraction (EF) or left ventricular diastolic dimension (EDD). The results demonstrated that TI and SDNN independently correlated with EF (multiple R = 0.59). Moreover, TI and SDNN independently correlated with EDD (multiple R = 0.45). CONCLUSION: TI and SDNN were indicators of the disease severity in myocardial dysfunction, while LF/HF, indicators of autonomic tone, did not have such an ability. It was of interest that the disease severity contributed to long-term fluctuations (TI, SDNN) of heart rate, rather than short-term fluctuations (LF/HF). PMID- 12848795 TI - Chronic inappropriate sinus tachycardia in elderly females. AB - BACKGROUND: The vast majority of reports of inappropriate sinus tachycardia (IST) had included predominantly young females with ages ranging between 15 and 46 years. In contrast, the present study presents the findings in four elderly females (aged 61-71 years) with long-standing symptoms of palpitations, more than 15 years, in whom IST was diagnosed in their 6th or 7th decade. METHODS: Clinical and laboratory examinations including electrocardiograms, echocardiograms, and heart rate variability studies were retrospectively reviewed in four of the nine elderly female patients with long-standing cardiac symptoms. Indices of heart rate variability were compared to four age-matched asymptomatic elderly females with no structural heart disease. RESULTS: The resting electrocardiograms were normal but one had voltage criteria for left ventricular hypertrophy. All traditional time- and frequency-domain indices were significantly decreased in the symptomatic elderly females as compared to the controls. Echocardiograms showed normal ejection fraction, and in three patients evidence of diastolic dysfunction and mild left ventricular hypertrophy. Blood pressures were elevated, with systolic blood pressure ranging from 170 to 190 and diastolic blood pressure ranging from 94 to 106. Anxiety disorders were diagnosed in all patients. CONCLUSIONS: This study demonstrates that IST can be present in a wide spectrum of patient populations, including elderly populations. It seems that some cases of IST can become chronic if not treated adequately. Apparently, earlier identification and treatment could have improved the quality of life of the patients. Traditional HRV indices may not be valuable to support either a sympathetic or parasympathetic mechanism. Therefore, further studies should consider the potential usefulness of the nonlinear method such as, for example, approximate entropy and detrended fluctuation analysis. PMID- 12848796 TI - Heart rate variability in obstructive sleep apnea: a prospective study and frequency domain analysis. AB - BACKGROUND: Cyclic variation of the heart rate is observed during apneic spells in obstructive sleep apnea (OSA). We hypothesized that autonomic changes would affect frequency-domain measures of heart rate variability (HRV). METHODS: We studied 20 patients (15 men, 5 women, mean age 47.2 +/- 12.2 years) with suspected OSA undergoing overnight polysomnography, and five patients (4 men, 1 woman, mean age 49.2 +/- 8.6 years) with recently diagnosed sleep apnea undergoing polysomnography while wearing continuous positive airway pressure (CPAP). Holter monitors were applied during sleep studies and data were analyzed in 5-minute blocks over the course of the night. Using spectral analysis, low frequency (LF) and high frequency (HF) powers were calculated for each interval. Overall mean and standard deviation (SD) for LF power, HF power, and the LF:HF ratio were recorded for each patient. Comparisons were made between patients with severe OSA (apnea hypopnea index (AHI) > 30, n = 8), moderate OSA (AHI 1-30, n = 5), without OSA (AHI < 10, n = 7), and patients wearing CPAP (n = 5). RESULTS: Assessment of overnight LF or HF power revealed no significant difference between the four groups. The LF:HF ratio, which represents sympathovagal balance, was higher among those with moderate disease compared to normals and those with severe OSA (both P = 0.037). The standard deviation of the LF:HF ratio was higher among those with moderate disease compared to normals (P = 0.0064) and those with severe OSA (P = 0.0006). OSA patients receiving CPAP behaved like patients with moderate OSA, with increased SD of the LF:HF ratio. CONCLUSIONS: The observed changes in the LF:HF ratio and its SD suggest an increased sympathetic tone and discordance in sympathovagal activity in moderate OSA, which is blunted in severe OSA. CPAP may restore autonomic defects, characteristic of severe OSA, to moderate levels. PMID- 12848797 TI - Effects of right coronary artery PTCA on variables of P-wave signal averaged electrocardiogram. AB - BACKGROUND: P-wave signal averaged ECG has been used to detect atrial late potentials that were found in paroxysmal atrial fibrillation. Ischemia is supposed to trigger ventricular late potentials, which indicate an elevated risk for ventricular tachycardia. Preexistent ventricular late potentials measured by ventricular signal averaged ECG is supposed to be eliminated by successful PTCA. METHODS: We examined the incidence of atrial late potentials in patients with a proximal stenosis of the right coronary artery and new onset of atrial fibrillation. Furthermore, we investigated the anti-ischemic effect of a successful percutaneous transluminal coronary angioplasty.(PTCA) of the right coronary artery. P-wave signal averaged ECG from 23 patients who had a PTCA of the right coronary artery (group A) were compared to age, sex, and disease matched control subjects (group B) one day before, one day after, and one month after PTCA. RESULTS: A new appearance of paroxysmal atrial fibrillation was presented in eight patients before PTCA (group A1) of group A. Patients with a stenosis of the right coronary artery had a significantly higher incidence of supraventricular extrasystoles in a 24-hour-Holter ECG (131.1 +/- 45.4 vs 17.1 +/ 18.9, P < 0.0002). The duration of the filtered P wave was longer (124.8 +/- 11.9 vs 118.5 +/- 10.1 ms, P < 0.04) and the root mean square of the last 20 ms (RMS 20) was significantly lower in group A than in group B (2.87 +/- 1.09 vs 3.97 +/- 1.12 micro V, P < 0.01). A successful PTCA caused an increase in RMS 20 (2.87 +/- 1.11 vs 4.19 +/- 1.19 microV, P < 0.02) and a decrease in filtered P wave duration (124.8 +/- 11.9 vs 118.4 +/- 10.4 ms, P < 0.04). Preexistent atrial late potentials were found among 15 patients before PTCA. After successful PTCA only 3 out of 15 patients were affected (P < 0.0004) after one day, as well as after one month. All patients with a history of atrial fibrillation did not suffer from an arrhythmic recurrence within the following six months after successful PTCA. CONCLUSION: A stenosis of the right coronary artery is associated with atrial late potentials. A successful PTCA of the right coronary artery eliminates preexistent atrial late potentials and may reduce the risk of atrial fibrillation. PMID- 12848798 TI - Knowledge deficits related to the QT interval could affect patient safety. AB - BACKGROUND: Recently, some QT-prolonging, noncardiac medications were withdrawn from the U.S. drug market because of continued inappropriate use by health care practitioners despite warnings and label changes from both the drug manufacturers and the U.S. Food and Drug Administration. This led us to assess the health care practitioners' knowledge of the QT interval and medications that may prolong it. METHODS: We surveyed health care practitioners, primarily specialists in cardiology, to identify knowledge deficits related to the QT interval. RESULTS: From a total of 334 survey respondents, 157 (47%) were physicians; 271 (81%) stated that cardiology was their area of specialization. Most of the respondents (86%) said that they would check an ECG before and after starting QT-prolonging medications, but less than half (42%) of all respondents and only 60% of physician respondents were able to accurately measure a sample QT interval on the survey. Less than two-thirds (63%) of respondents were able to accurately identify possible QT-prolonging medications, while only about half (51%) could accurately identify medication combinations that might prolong the QT interval. CONCLUSIONS: We identified significant knowledge deficits regarding the QT interval and QT-prolonging medications. Additional research is needed to determine the extent to which these knowledge deficits may negatively affect patient safety. We must also develop effective strategies to eliminate these deficits. PMID- 12848799 TI - Noninvasive risk stratification in arrhythmogenic right ventricular cardiomyopathy. AB - The natural history of arrhythmogenic right ventricular cardiomyopathy is determined by the electrical instability of the dystrophic myocardium, which can precipitate arrhythmic cardiac arrest any time during the course of the disease and by the progressive myocardial loss that results in ventricular dysfunction and heart failure. Sudden death accounts for the majority of the fatal events but its occurrence is mostly unpredictable. There are no prospective and controlled studies assessing clinical markers that can predict the occurrence of life threatening ventricular arrhythmias. However, the noninvasive risk profile, which emerges from retrospective analysis of clinical and pathologic series, is characterized by history of syncope, physical exercise, spontaneous ventricular tachycardia or ventricular fibrillation, right ventricular dysfunction, left ventricular involvement, right precordial negative T wave, right bundle branch block, QT-QRS dispersion, right precordial ST-segment elevation and late potentials. At present only QRS dispersion, history of syncope and right and/or left ventricular abnormalities at radionuclide angiography proved to be independent noninvasive predictors of sudden death. PMID- 12848800 TI - Introductory note to a classic article by Francis Galton. PMID- 12848801 TI - A diagram of heredity. PMID- 12848805 TI - Atrial fibrillation. PMID- 12848802 TI - An unusual cause of reflex cardiovascular syncope: vagal paraganglioma. AB - Vagal paraganglioma is one of the rare tumors of the neuroendocrine system. We are reporting a vagal paraganglioma case presented with uncommon features of the disease: asystole and syncope. Syncope episodes occurred 3 years before the major symptoms of the disease. Dual chamber pacemaker failed to prevent syncope attacks because of the vasodepressor component. The patient was treated successfully with en bloc removal of tumor and vagal nerve. Syncope episodes disappeared after operation. PMID- 12848806 TI - White blood cell count and the occurrence of silent ischemia after myocardial infarction. AB - BACKGROUND: Inflammation plays a role in the pathogenesis of atherosclerosis. Attempts are made to use markers of inflammation as prognostic factors in coronary artery disease and acute coronary syndromes. The correlation between inflammation and silent postinfarction ischemia is unknown. METHODS: The study population consists of 104 asymptomatic patients who had uncomplicated Q-wave myocardial infarction within 6 months prior to the enrollment. After the white blood cell (WBC) count was assessed, the population was divided into two groups: group I comprising 48 patients with WBC < or = 7.0 x 103/microl and group II comprising 56 patients with WBC > 7.0 x 103/microl. Twenty-four-hour Holter monitoring was performed to detect the presence of silent ischemia. RESULTS: Eighty-eight silent ischemic episodes were recorded. Ischemia on Holter monitoring was detected in 47 patients (84%) from group II and in five patients (9%) in group I (P < 0.01). We have found a significant positive correlation between WBC count and the number of ischemic episodes (r = 0.25), their maximal amplitude (r = 0.39), duration (r = 0.34), and total ischemic burden (r = 0.36). In multivariate analysis leucocytosis proved to be the only parameter independently correlated with the presence of silent ischemia. CONCLUSION: Postinfarction asymptomatic patients with increased WBC count are more likely to have residual ischemia. PMID- 12848807 TI - Seasonal variability of QT dispersion in healthy young males. AB - BACKGROUND: There are few data related to the seasonal influences on the QT dispersion. METHODS: We analyzed the effects of seasons on QT dispersion in a large group of healthy young males. We studied the seasonal variability of QT dispersion in 523 healthy male subjects aged 22 +/- 4 years (ranging from 20 to 26). Four seasonal 12-lead resting electrocardiograms (ECGs) recorded at double amplitude were performed at 25 mm/s at intervals of 3 months. Subsequent ECGs were recorded within 1 hour of the reference winter recording. QT dispersion was defined as the difference between the longest and the shortest mean QT intervals. RESULTS: There was a significant seasonal variation in QT dispersion (P = 0.001), with the largest QT dispersion in winter (71 +/- 18 ms) and the smallest one in spring (43 +/- 19). CONCLUSION: There exists a significant seasonal variation in QT dispersion of healthy subjects and such variability should be taken into consideration in the evaluation process of QT dispersion. PMID- 12848808 TI - Sildenafil citrate does not affect QT intervals and QT dispersion: an important observation for drug safety. AB - BACKGROUND: Sildenafil is an effective and widely used therapeutic agent for erectile dysfunction. Deaths have been reported due to sildenafil use and most of them are attributed to concurrent use of nitrates. However, the effects of sildenafil on QT intervals, QT dispersion, and the possible risk of ventricular arrhythmia have not been studied before. Our aim in this study was to evaluate the effect of sildenafil citrate on QT intervals and QT dispersion. METHODS: Thirty-six patients with erectile dysfunction were included in this study. Twenty one patients had coronary artery disease whereas 12 of them also had accompanying diabetes mellitus. Standard 12-lead electrocardiograms (ECG) were recorded three times: before, and at the first and fourth hours of 50 mg sildenafil citrate ingestion. All QT parameters were corrected for heart rate. RESULTS: Mean age of the patients was 54 +/- 12 years. The mean heart rate did not differ significantly between the three ECG examinations. The corrected and uncorrected maximum and minimum QT intervals were not significantly different between the three ECG examinations. The QT dispersion and corrected QT dispersion before and 1 hour and 4 hours after sildenafil ingestion were 31 +/- 9 ms, 36 +/- 10 ms; 32 +/- 11 ms, 37 +/- 14 ms; 27 +/- 8 ms, 32 +/- 9 ms, respectively (P > 0.05). CONCLUSIONS: Sildenafil does not prolong QT intervals or increase QT dispersion in patients with erectile dysfunction. Our results suggest that the risk of ventricular arrhythmia does not increase with ingestion of 50 mg sildenafil. PMID- 12848809 TI - Increased p-wave duration and p-wave dispersion in patients with aortic stenosis. AB - BACKGROUND: P-wave dispersion (PWD), defined as the difference between the maximum and minimum P-wave duration, has been proposed as being useful for the prediction of paroxysmal atrial fibrillation (AF). AF is the most common arrhythmia and an important prognostic indicator for clinical deterioration in patients with aortic stenosis (AS). The aim of the present study was to evaluate PWD in patients with AS. METHODS: The study population consisted of two groups: Group I consisted of 98 patients with AS (76 men, 22 women; aged 63 +/- 8 years) and group II consisted of 98 healthy subjects (same age and sex) without any cardiovascular disease. A 12-lead electrocardiogram was recorded for each subject. The P-wave duration was calculated in all leads of the surface electrocardiogram. The difference between the maximum and minimum P-wave duration was calculated and was defined as the PWD. All patients and control subjects were also evaluated by echocardiography to measure the left atrial diameter, left ventricular ejection fraction, left ventricular wall thicknesses, and the maximum and mean aortic gradients. Patients were also evaluated for the presence of paroxysmal AF. RESULTS: Maximum P-wave duration and PWD of group I were found to be significantly higher than those of group II. In addition, patients with paroxysmal AF had significantly higher PWD than those without paroxysmal AF. There was no significant difference between the two groups regarding minimum P wave duration. In addition, there was no significant correlation between echocardiographic variables and PWD. CONCLUSION: PWD, indicating increased risk for paroxysmal AF, was found to be significantly higher in patients with AS than in those without it. Further assessment of the clinical utility of PWD for the prediction of paroxysmal AF in patients with severe AS will require longer prospective studies. PMID- 12848810 TI - The relation of QT dispersion and localized QT difference to coronary pathology in a population with unstable coronary artery disease. AB - BACKGROUND: QT dispersion (QTd) contains prognostic information in several patient groups. The variable increases in several conditions with ischemia. Originally, it was thought to reflect the local repolarization inhomogeneity. Even though this explanation has been questioned lately, it continues to be put forward. In order to elucidate a possible local mechanism, we investigated the relation between QT dispersion, an ECG parameter reflecting the local dispersion, and angiographical measures in a population with unstable coronary artery disease. METHODS: The 276 patients were recruited from the FRISC II trial. As the QTd parameter we used the mean value of automatically measured QTd during 27 hours after admission (QTdMean). As a local repolarization measure we used the maximal difference in QT between two adjacent ECG leads (QTdiffMean). The computations were performed on all available ECG leads and on a restricted set without the V1-V2 combination. Previously published angiographic scoring tools were adapted for rating and localizing the coronary pathology by two approaches and applied on 174 patients undergoing angiography. RESULTS: QTdMean was significantly higher than that reported in previous material with unselected chest pain patients (55 vs 40 ms). QTdiffMean correlated strongly with QTdMean. No differences in QTdMean were detected between patients with different angiographical scores. No relation could be shown between the region with dominating coronary pathology as expressed by the scoring tools and the localization of QTdiffMean. CONCLUSIONS: QTd in ischemia seems to be increased by a mechanism unrelated to localization and severity of coronary disease. PMID- 12848811 TI - Relationship between ST-segment morphology and conduction disturbances detected by signal-averaged electrocardiography in Brugada syndrome. AB - BACKGROUND: Although arrhythmogenesis of Brugada syndrome is still unknown, it has been reported to be associated with conduction disturbances. Two ST-segment morphologies (coved and saddle-back patterns) have been described in this syndrome. No study has sought to determine which morphology has stronger conduction disturbances, thereby associating with life-threatening events. METHODS: Forty-six patients who presented the Brugada-type ECG with either of a characteristic coved (n = 25) or saddle-back (n = 21) pattern of ST-segment morphology underwent signal-averaged ECG (SAECG). SAECG parameters, and the history of life-threatening events defined as syncope or aborted sudden death, were compared between groups. RESULTS: Although filtered QRS duration did not differ between groups, the incidence of late potentials in the coved group was higher than in the saddle-back group (22 patients (88%) versus 4 patients (19%); P < 0.01), showing lower RMS40 and longer LAS40. Life-threatening events occurred in 17 patients (68%) in the coved group and 7 patients (33%) in the saddle-back group (P = 0.02). CONCLUSION: The coved pattern of ST segment was more closely related to conduction disturbances than the saddle-back pattern in patients with Brugada-type ECG. Life-threatening events were more common in patients with the coved ST-segment elevation. Conduction disturbances in the coved pattern of ST segment may reflect a substrate of arrhythmogenesis in Brugada syndrome. PMID- 12848812 TI - Prevalence of potential noninvasive arrhythmia risk predictors in healthy, middle aged persons. AB - BACKGROUND: To date, prevalence and clinical significance of noninvasive arrhythmia risk predictors in apparently healthy, middle-aged persons are largely unknown. METHODS: A total of 110 apparently healthy persons 20-75 years old were enrolled in this prospective observational monocenter study and followed up for 32 +/- 15 months. Baseline investigations included symptom-limited bicycle ergometry, echocardiography, time-domain analysis, and spectral turbulence analysis of the signal-averaged electrocardiogram (ECG), ventricular arrhythmias, and heart rate variability on 24-hour Holter ECG, baroreflex sensitivity, and t wave alternans in all persons. RESULTS: The prevalence of an abnormal signal averaged ECG was 1% for spectral turbulence analysis and varied between 1% and 37% for time-domain analysis depending upon the definition used for an abnormal time-domain analysis. A reduced heart rate variability defined as a standard deviation of normal-to-normal intervals < or =105 ms, <100 ms and <70 ms was found in 12%, 9%, and 1% of persons. A baroreflex sensitivity <6 ms/mmHg and <3 ms/mmHg was present in 15% and 2% of persons. Microvolt t-wave alternans was found to be positive in 5%, negative in 88%, and indeterminate in 7% of persons, respectively. During the 32 +/- 15 months follow-up, no arrhythmic events and no cardiovascular mortality were observed in this population. CONCLUSIONS: Abnormal findings of noninvasive arrhythmia risk stratification can be found in 1-37% of healthy, middle-aged persons when previously reported cut-off values are used. PMID- 12848813 TI - Long-term improvement of QT dispersion is unaffected by short-term changes in blood pressure during treatment of systemic hypertension with enalapril. AB - BACKGROUND: We report the reduction of QT and QTc dispersion in patients treated for 7 years with enalapril for systemic hypertension with left ventricular (LV) hypertrophy. We assess the correlation between QT dispersion and LV mass during this period and at the end of an 8-week period of suspension of enalapril treatment after 5 years. METHODS: Twenty-four previously untreated patients with this condition took enalapril (20 mg twice daily) for 7 years, except during an 8 week period following 5-year follow-up. Cardiovascular parameters were determined by two-dimensional guided M-mode echocardiography, and QT interval was measured, in a pretreatment placebo phase, 8 weeks and 1, 3, 5, and 7 years after the start of the therapy, at the end of the 8-week suspension effected after 5 years, and 8 weeks after the end of the suspension. RESULTS: Therapy rapidly reduced blood pressure (BP) from 156/105 mmHg to normal values: 134/84 mmHg after 8 weeks' treatment, 130-84 mmHg at 7-year follow-up (P < 0.001 with respect to the placebo phase). LV mass index decreased progressively until at 5-year follow-up the reduction had reached 39% (P < 0.001), after which neither LV mass nor any other structural parameter underwent any further significant change. During this time, QT dispersion (DeltaQT) and the dispersion of "corrected" QT (DeltaQTc) decreased significantly: DeltaQT (from 61 +/- 21 to 37 +/- 13 ms) and DeltaQTc (from 67 +/- 27 to 41 +/- 16 ms). After suspension of treatment for 8 weeks following 5-year follow-up, DeltaQT was 40 +/- 14 ms and DeltaQTc was 44 +/- 17 ms; there were no significant changes either in DeltaQT and DeltaQTc or LV hypertrophy although BP had returned to pretreatment values (BP: 150 +/- 16; 101 +/- 10 mmHg). CONCLUSIONS: Long-term enalapril treatment of hypertensive patients with LV hypertrophy induces marked regression of LV mass and improvement of QT dispersion. These improvements occur on a longer timescale than improvement in BP, and are not affected by transient changes in BP values. PMID- 12848814 TI - Testing of a new T-wave subtraction algorithm as an aid to localizing ectopic atrial beats. AB - BACKGROUND: Identifying the timing and morphology of an ectopic P wave from the surface electrogram can aid in the diagnosis and localization of atrial arrhythmias. Given the relatively short coupling interval of atrial ectopic beats, the P wave is often obscured by the larger amplitude QRS-T wave complex. A method to uncover such "buried" P waves using a standard 12-lead surface ECG would be clinically useful and could potentially be a noninvasive guide to catheter ablation of focal atrial tachycardia. METHODS: We developed an automated computerized program (BARD DUO LAB SYSTEM trade mark ) designed to subtract the QRS-T wave complex from the surface electrogram and uncover a previously obscured P wave. The purpose of the present study was to validate this program. The surface ECG from 21 patients undergoing atrial pacing during electrophysiologic study (group I) and 10 patients with atrial tachycardia (group II) were analyzed and the derived P-wave morphology assessed using correlation waveform analysis (CWA) and visual grading by three reviewers. RESULTS: The algorithm successfully uncovered the P wave in each surface ECG. For the 21 patients in group I, average CWA comparing the derived P wave with the previous paced P wave was 83%. Average CWA for group II was 82%. Visual grading of the match between derived P waves and paced P waves revealed a 21/21 match in group I patients and a 12/12 match in 9/10 of group II patients. CONCLUSIONS: An ectopic atrial P wave obscured by a coincident QRS-T wave complex can be accurately uncovered using this new algorithm. Addition of this technique to existing methods may improve the diagnosis of atrial arrhythmias and aid in the localization and ablation of ectopic atrial foci. PMID- 12848815 TI - The predictive value of exercise QRS duration changes for post-PTCA coronary events. AB - BACKGROUND: The sensitivity and predictive values of exercise ECG testing using ST-T criteria after percutaneous transluminal coronary angioplasty (PTCA) are low, precluding its routine use for screening for restenosis. The predictive value of QRS duration criteria during exercise testing (ET) ECG after PTCA for future coronary events has not been reported. The aim of the study was to compare QRS duration changes with ST-T criteria during ET, as a predictor of coronary events after PTCA. METHODS: A prospective study of 206 consecutive patients who underwent ET at a mean of 34 +/- 14 days after their first PTCA, and were the followed for a mean of 23 +/- 9 months. Patients were divided by QRS duration into two groups-Q1: ischemic response (QRS duration prolongation of more than 3 ms relative to the resting duration), and Q2: normal response (QRS duration shortening or without change from resting duration). Patients were also divided by their ST-T response, S1: ischemic response, and S2: normal response. RESULTS: During follow-up 52 patients (58%) experienced restenosis or MI, or underwent CABG-Q1: 44 (85%), Q2: 8(15%) (P < 0.0002), S1: 8 (15%), S2: 44 (85%), (P < 0.641), two patients died-Q1: 1 (1%) and Q2: 1 (1%). For QRS and ST-T, the relative risk of having at least one of the coronary events was 4.02 (CI 2.1-9.9) versus 1.13 (CI 0.8-2.9), respectively. The sensitivity for future coronary events was 85% and 52% and the specificity was 48% and 98% for the QRS and ST-T criteria, respectively. CONCLUSION: QRS prolongation during peak ET ECG after PTCA is a more sensitive marker than ST-T criteria for detection of patients at risk for later coronary events. PMID- 12848816 TI - Usefulness of risk stratification for future cardiac events in infarct survivors with severely depressed versus near-normal left ventricular function: results from a prospective long-term follow-up study. AB - BACKGROUND: Although primary preventive therapy with implantable cardioverter defibrillators has recently been shown to be effective in patients with coronary artery disease and left ventricular dysfunction, further identification of patients at particularly high risk for arrhythmic death would improve the cost effectiveness of device therapy. The value of risk stratification in postinfarction patients with versus those without left ventricular dysfunction has not been investigated in detail in infarct survivors treated according to contemporary therapeutic guidelines. METHODS: Patients with acute myocardial infarction underwent coronary angiography including left ventricular angiography in an attempt to restore antegrade flow of the infarct-related artery. Additionally, patients underwent noninvasive autonomic risk stratification by means of heart rate variability (HRV) and baroreflex sensitivity (BRS) measurements prior to hospital discharge. RESULTS: A total of 411 patients were prospectively included in the study. The primary study endpoint of cardiac death and arrhythmic events was significantly more common in patients with LVEF < or = 35% as compared to those with preserved LV function (27% vs 4%; P < 0.0001). In patients with LV dysfunction, HRV and BRS were significant risk predictors on univariate (P < 0.01 for BRS; P = 0.04 for HRV) and multivariate (P = 0.028 for BRS; P = 0.053 for HRV) analyses. In contrast, in patients with preserved LV function, only patency of the infarct artery but not autonomic markers was significantly predictive of cardiac death and arrhythmic events. CONCLUSION: The present study demonstrates that autonomic testing does not yield predictive power in infarct survivors with preserved left ventricular function. Accordingly, cost effectiveness of risk stratification and subsequent preventive therapy may be improved by restricting risk stratification to patients with impaired LV function. PMID- 12848817 TI - Sympathetic nervous system activity and ventricular tachyarrhythmias: recent advances. AB - Sympathetic nervous system activity (SNSA) is believed to participate in the genesis of ventricular tachyarrhythmias (VTA) but understanding has been impeded by the number and complexity of effects and the paucity of data from humans. New information from studies of genetic disorders, animal models, and spontaneous human arrhythmias indicates the importance of the temporal pattern of SNSA in arrhythmia development. The proarrhythmic effects of short-term elevations of SNSA are exemplified by genetic disorders and include enhancement of early and delayed afterdepolarizations and increased dispersion of repolarization. The role of long-term elevations of SNSA is suggested by animal models of enhanced SNSA signaling that results in apoptosis, hypertrophy, and fibrosis, and sympathetic nerve sprouting caused by infusion of nerve growth factor. Processes that overlap short- and long-term effects are suggested by changes in R-R interval variability (RRV) that precede VTA in patients by several hours. SNSA-mediated alterations in gene expression of ion channels may account for some intermediate-term effects. The propensity for VTA is highest when short-, intermediate, and long-term changes are superimposed. Because the proarrhythmic effects are related to the duration and intensity of SNSA, normal regulatory processes such as parasympathetic activity that inhibits SNSA, and oscillations that continuously vary the intensity of SNSA may provide vital antiarrhythmic protection that is lost in severe heart failure and other disorders. These observations may have therapeutic implications. The recommended use of beta-adrenergic receptor blockers to achieve a constant level of inhibition does not take into account the temporal patterns and regional heterogeneity of SNSA, the proarrhythmic effects of alpha-adrenergic receptor stimulation, or the potential proarrhythmic effects of beta-adrenergic receptor blockade. Further research is needed to determine if other approaches to SNSA modulation can enhance the antiarrhythmic effects. PMID- 12848818 TI - History of atrial fibrillation. PMID- 12848819 TI - Auricular flutter and fibrillation. 1911. PMID- 12848820 TI - Predicting atrial fibrillation after mitral valve replacement. PMID- 12848821 TI - An Arabidopsis porB porC double mutant lacking light-dependent NADPH:protochlorophyllide oxidoreductases B and C is highly chlorophyll-deficient and developmentally arrested. AB - A key reaction in the biosynthesis of chlorophylls (Chls) a and b from cyanobacteria through higher plants is the strictly light-dependent reduction of protochlorophyllide (Pchlide) a to chlorophyllide (Chlide) a. Angiosperms, unlike other photosynthetic organisms, rely exclusively upon this mechanism to reduce Pchlide and hence require light to green. In Arabidopsis, light-dependent Pchlide reduction is mediated by three structurally related but differentially regulated NADPH:Pchlide oxidoreductases, denoted as PORA, PORB, and PORC. The PORA and PORB genes, but not PORC, are strongly expressed early in seedling development. In contrast, expression of PORB and PORC, but not PORA, is observed in older seedlings and adult plants. We have tested the hypothesis that PORB and PORC govern light-dependent Chl biosynthesis throughout most of the plant development by identifying porB and porC mutants of Arabidopsis, the first higher plant por mutants characterized. The porB-1 and porC-1 mutants lack the respective POR transcripts and specific POR isoforms because of the interruption of the corresponding genes by a derivative of the maize Dissociation (Ds) transposable element. Single por mutants, grown photoperiodically, display no obvious phenotypes at the whole plant or chloroplast ultrastructural levels, although the porB-1 mutant has less extensive etioplast inner membranes. However, a light grown porB-1 porC-1 double mutant develops a seedling-lethal xantha phenotype at the cotyledon stage, contains only small amounts of Chl a, and possesses chloroplasts with mostly unstacked thylakoid membranes. PORB and PORC thus seem to play redundant roles in maintaining light-dependent Chl biosynthesis in green plants, and are together essential for growth and development. PMID- 12848822 TI - cPrG-HCl a potential H+/Cl- symporter prevents acidification of storage vacuoles in aleurone cells and inhibits GA-dependent hydrolysis of storage protein and phytate. AB - The putative H+/Cl- symporter cycloprodigiosin-HCl (cPrG-HCl) was used to investigate the role of vacuole acidification in cereal aleurone cell function. The protein storage vacuole (PSV) becomes acidified rapidly when aleurone cells are treated with gibberellic acid (GA) but not abscisic acid (ABA). We show that cPrG prevents PSV acidification in aleurone layers and prevents synthesis of secretory proteins such as alpha-amylase. Our data support the hypothesis that decreased hydrolase synthesis is a consequence of decreased hydrolysis of storage proteins in PSV. Support for this hypothesis comes from experiments showing that breakdown of barley 7S globulins and phytate is inhibited by cPrG in GA-treated aleurone layers. Decreased mobilization of PSV reserves is accompanied by reductions in the free amino acid pool size and in the amount of ions released from the aleurone layer. Vacuolation of the aleurone cell is a diagnostic feature of the response to GA, and vacuolation is also inhibited by cPrG. Evidence that cPrG acts as a potential H+/Cl- symporter in aleurone is presented. We show that cPrG does not inhibit the synthesis and secretion of alpha-amylase when Cl- ions are omitted from the incubation medium. Although cPrG blocks many GA-induced responses of aleurone layers, it does not affect early steps in GA signaling. The SLN1 protein, a negative regulator of GA signaling, is turned over in GA-treated cells in the presence and absence of cPrG. Similarly, synthesis of the transcriptional activator GAMYB is unaffected by the presence of cPrG in GA treated cells. PMID- 12848823 TI - Functional expression of AtHMA4, a P1B-type ATPase of the Zn/Co/Cd/Pb subclass. AB - Mechanisms are required by all organisms to maintain the concentration of essential heavy metals (e.g. Zn and Cu) within physiological limits and to minimise the detrimental effects of non-essential heavy metals (e.g. Cd). Heavy metal P-type ATPases (HMAs) are a subgroup of the P-type ATPase superfamily that may contribute to metal homeostasis in plants. We cloned and characterised a member of this family, AtHMA4, from Arabidopsis thaliana that clusters with the Zn/Co/Cd/Pb subclass of HMAs on phylogenetic analysis. Sequencing of the AtHMA4 cDNA showed that it contained the conserved motifs found in all P-type ATPases and also motifs that are characteristic of heavy-metal ATPases. Escherichia coli mutants defective in the HMAs, CopA and ZntA, were used in functional complementation studies. AtHMA4 was able to restore growth at high [Zn] in the zntA mutant but not at high [Cu] in the copA mutant, suggesting a role in zinc transport. Heterologous expression of AtHMA4 in Saccharomyces cerevisiae made the yeast more resistant to Cd but did not affect sensitivity to other metals compared with vector-transformed controls. The organ specificity of AtHMA4 was analysed in Arabidopsis and showed that AtHMA4 was expressed in a range of tissues with highest expression in roots. AtHMA4 was upregulated in roots exposed to elevated levels of Zn and Mn but downregulated by Cd. Possible physiological roles of this transporter in Arabidopsis are discussed. PMID- 12848824 TI - AtMYB103 regulates tapetum and trichome development in Arabidopsis thaliana. AB - The AtMYB103 gene is a member of the R2R3 MYB gene family in Arabidopsis thaliana. Using the GUS reporter gene, AtMYB103 expression was found to be restricted to the tapetum of developing anthers. Employing RT-PCR and in situ hybridisation, we now show that AtMYB103 is also expressed in trichomes. GUS expression in trichomes was obtained by incorporating the coding and 3' untranslated regions of AtMYB103 into the promoter-GUS constructs. Sense and antisense technologies were used to downregulate AtMYB103 expression. In transgenic lines with reduced AtMYB103 transcript levels, pollen, tapetum and trichome development were altered. The majority of the pollen grains were distorted in shape and had reduced or no cytoplasmic content. Tapetal degeneration occurred early, and large opaque bodies appeared in the tapetal cytoplasm. In transgenic plants, trichomes on cauline and rosette leaves produced additional branches. These overbranched trichomes contained more nuclear DNA than the wild-type trichomes. The results indicate that AtMYB103 is required for tapetal development and microsporogenesis, and negatively regulates trichome endoreduplication linked to the trichome branching. PMID- 12848825 TI - Arabidopsis local resistance to Botrytis cinerea involves salicylic acid and camalexin and requires EDS4 and PAD2, but not SID2, EDS5 or PAD4. AB - Salicylic acid (SA) is an important regulator of plant defense responses, and a variety of Arabidopsis mutants impaired in resistance against bacterial and fungal pathogens show defects in SA accumulation, perception, or signal transduction. Nevertheless, the role of SA-dependent defense responses against necrotrophic fungi is currently unclear. We determined the susceptibility of a set of previously identified Arabidopsis mutants impaired in defense responses to the necrotrophic fungal pathogen Botrytis cinerea. The rate of development of B. cinerea disease symptoms on primary infected leaves was affected by responses mediated by the genes EIN2, JAR1, EDS4, PAD2, and PAD3, but was largely independent of EDS5, SID2/ICS1, and PAD4. Furthermore, plants expressing a nahG transgene or treated with a phenylalanine ammonia lyase (PAL) inhibitor showed enhanced symptoms, suggesting that SA synthesized via PAL, and not via isochorismate synthase (ICS), mediates lesion development. In addition, the degree of lesion development did not correlate with defensin or PR1 expression, although it was partially dependent upon camalexin accumulation. Although npr1 mutant leaves were normally susceptible to B. cinerea infection, a double ein2 npr1 mutant was significantly more susceptible than ein2 plants, and exogenous application of SA decreased B. cinerea lesion size through an NPR1-dependent mechanism that could be mimicked by the cpr1 mutation. These data indicate that local resistance to B. cinerea requires ethylene-, jasmonate-, and SA-mediated signaling, that the SA affecting this resistance does not require ICS1 and is likely synthesized via PAL, and that camalexin limits lesion development. PMID- 12848826 TI - The lower cell density of leaf parenchyma in the Arabidopsis thaliana mutant lcd1 1 is associated with increased sensitivity to ozone and virulent Pseudomonas syringae. AB - Under optimal growth conditions (120 micro mol photons m-2 sec-1 photosynthetically active radiation (PAR), 16-h photoperiod), the recessive ozone sensitive Arabidopsis thaliana L. Heynh. mutant lcd1-1 exhibits a pale phenotype compared to the wild type. Confocal and multiphoton microscopy revealed that the paleness of lcd1-1 is because of a lower cell density in the leaf palisade parenchyma, resulting in decreased chlorophyll content. When exposed to ozone, lcd1-1 leaves become paler and contain an increased amount of the lipid peroxidation product malondialdehyde compared to the wild type, suggesting that lcd1-1 suffers from elevated levels of reactive oxygen species (ROS) generated in the apoplast. Infection of leaves with virulent Pseudomonas syringae reveals higher bacterial growth as well as lower pathogenesis-related protein 1 (PR-1) and PR-5 expression in lcd1-1 than in the wild type. When the wild type and lcd1 1 are exposed to short-term high-light stress, leaves do not bleach in lcd1-1 and potential activities of photosystems I (PSI) and II (PSII) decrease to a similar extent in both the genotypes, indicating that the photosynthetic apparatus is not affected by lcd1-1 mutation. The LCD1 gene, found to contain a nonsense mutation in the mutant, has been identified. It is located at the bottom of chromosome 2 of the Arabidopsis genome. However, the function of the protein encoded by LCD1 is not yet known. We hypothesize that LCD1 plays a role in normal leaf development, and that the increased sensitivity to ozone and virulent P. syringae is a secondary effect that presumably results from the lower-cell-density phenotype in lcd1-1. PMID- 12848827 TI - Transfer of T-DNA and Vir proteins to plant cells by Agrobacterium tumefaciens induces expression of host genes involved in mediating transformation and suppresses host defense gene expression. AB - Agrobacterium tumefaciens is a plant pathogen that incites crown gall tumors by transferring to and expressing a portion of a resident plasmid in plant cells. Currently, little is known about the host response to Agrobacterium infection. Using suppressive subtractive hybridization and DNA macroarrays, we identified numerous plant genes that are differentially expressed during early stages of Agrobacterium-mediated transformation. Expression profiling indicates that Agrobacterium infection induces plant genes necessary for the transformation process while simultaneously repressing host defense response genes, thus indicating successful utilization of existing host cellular machinery for genetic transformation purposes. A comparison of plant responses to different strains of Agrobacterium indicates that transfer of both T-DNA and Vir proteins modulates the expression of host genes during the transformation process. PMID- 12848828 TI - Identification of regulatory sequence elements within the transcription promoter region of NpABC1, a gene encoding a plant ABC transporter induced by diterpenes. AB - Expression of NpABC1, a gene encoding a plasma membrane ATP binding cassette (ABC) transporter in Nicotiana plumbaginifolia, is induced by sclareol, an antifungal diterpene produced at the leaf surface, as well as by sclareolide, a close analog. A genomic fragment including the 1282-bp region upstream of the NpABC1 transcription start was fused to the reporter beta-glucuronidase (gus) gene and introduced into N. tabacum BY2 cells for stable transformation. A 25 fold increase in gus expression was observed when cells were treated with sclareolide and some other terpenes. The combined use of 5'-deletion promoter analysis, gel mobility shift assays, DNase I footprinting, and site-directed mutagenesis allowed us to identify three cis-elements (sclareol box 1 (SB1), SB2, and SB3) located, respectively, within nucleotides -827 to -802, -278 to -243, and -216 to -190 upstream of the NpABC1 transcription start. In vivo evaluation of these elements on sclareolide-induced expression showed that mutation of SB1 reduced expression by twofold, while that of SB2 had no effect. On the other hand, SB3 had a marked effect as it completely abolished sclareolide-mediated expression. NpABC1-gus expression was not induced by the stress signals, salicylic acid and ethylene, but was mediated, to some extent, by methyl jasmonate, which is known to promote diterpene synthesis. PMID- 12848829 TI - In vitro analysis of the sequences required for transcription of the Arabidopsis thaliana 5S rRNA genes. AB - In vivo, we have already shown that only two of the 5S rDNA array blocks of the Arabidopsis thaliana genome produce the mature 5S rRNAs. Deletions and point mutations were introduced in an Arabidopsis 5S rDNA-transcribed region and its 5' and 3'-flanks in order to analyse their effects on transcription activity. In vitro transcription revealed different transcription control regions. One control region essential for transcription initiation was identified in the 5'-flanking sequence. The major sequence determinants were a TATA-like motif (-28 to -23), a GC dinucleotide (-12 to -11), a 3-bp AT-rich region (-4 to -2) and a C residue at -1. They are important for both accurate transcription initiation and transcription efficiency. Transcription level was regulated by polymerase III (Pol III) re-initiation rate as in tRNA genes in which TATA-like motif is involved. Active 5S rDNA transcription additionally required an intragenic promoter composed of an A-box, an Intermediate Element (IE) and a C-box. Double stranded oligonucleotides corresponding to different fragments of the transcribed region, used as competitors, revealed the main importance of internal promoter elements. A stretch of four T is sufficient for transcription termination. Transcription of Arabidopsis 5S rDNA requires 30 bp of 5'-flanking region, a promoter internal to the transcribed region, and a stretch of T for transcription termination. PMID- 12848830 TI - The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats. AB - A single radish nuclear gene, Rfo, restores Ogura (ogu) cytoplasmic male sterility (CMS) in Brassica napus. A map-based cloning approach relying on synteny between radish and Arabidopsis was used to clone Rfo. A radish gene encoding a 687-amino-acid protein with a predicted mitochondrial targeting pre sequence was found to confer male fertility upon transformation into ogu CMS B. napus. This gene, like the recently described Petunia Rf gene, codes for a pentatricopeptide repeat (PPR)-containing protein with multiple, in this case 16, PPR domains. Two similar genes that do not appear to function as Rfo flank this gene. Comparison of the Rfo region with the syntenic Arabidopsis region indicates that a PPR gene is not present at the Rfo-equivalent site in Arabidopsis, although a smaller and related PPR gene is found about 40 kb from this site. The implications of these findings for the evolution of restorer genes and other PPR encoding genes are discussed. PMID- 12848831 TI - Gene trapping of the Arabidopsis genome with a firefly luciferase reporter. AB - Experiments with gene-trap vectors containing the firefly luciferase (LUC) reporter genes were carried out with the aim of analyzing functions of the Arabidopsis genome. Studies with protein fusion-type trap vectors as well as an internal ribosome entry site (IRES)-assisted non-fusion-type vector revealed that both types of vectors were suitable for gene trapping in Arabidopsis, although there were some differences in trapping efficiencies. The established trap lines were subjected to analyses for light responses, demonstrating the powerful and unique applications of a LUC-trapping system. A systematic survey of the insertion sites of the T-DNAs in LUC-expressing lines revealed 12-41% gene trapping efficiencies depending on the vector. We demonstrate that the LUC trapping system provides a unique system with which to monitor temporal expression of plant genes. PMID- 12848832 TI - Psychosocial intervention for rural women with breast cancer: The Sierra-Stanford Partnership. AB - OBJECTIVE: This study was initiated by breast cancer survivors living in a rural community in California. They formed a partnership with academic researchers to develop and evaluate a low-cost, community-based Workbook-Journal (WBJ) for improving psychosocial functioning in geographically and economically isolated women with primary breast cancer. DESIGN: A randomized controlled trial was used to compare the WBJ intervention plus educational materials to educational materials alone (usual care). SETTING: One rural cancer center and several private medical, surgical, and radiation oncology practices in 7 rural counties in the Sierra Nevada Foothills of California. PARTICIPANTS: One hundred women with primary breast cancer who were either within 3 months of diagnosis or within 3 months of completing treatment. INTERVENTION: A community-initiated, theoretically-based Workbook-Journal, designed by rural breast cancer survivors and providers as a support group alternative. It included compelling personal stories, local rural resources, coping strategies, and messages of hope. RESULTS: Community recruiters enrolled 83% of the women referred to the study. Retention at 3-month follow-up was 98%. There were no main effects for the WBJ. However, 3 significant interactions suggested that women who were treated in rural practices reported decreased fighting spirit and increased emotional venting and posttraumatic stress disorder symptoms if they did not receive the WBJ. Among women who receive the WBJ, 74% felt emotionally supported. CONCLUSIONS: This community-based Workbook-Journal may be an effective psychosocial intervention for rural, isolated, and low-income women with breast cancer. Community involvement was essential to the success of this project. PMID- 12848833 TI - Identifying older Chinese immigrants at high risk for osteoporosis. AB - BACKGROUND: Data about whether Asian Americans are a high-risk or a low-risk group for osteoporosis are limited and inconsistent. Few previous studies have recognized that the heterogeneity of the Asian American population, with respect to both nativity (foreign- vs U.S.-born) and ethnicity, may be related to osteoporosis risk. OBJECTIVE: To assess whether older foreign-born Chinese Americans living in an urban ethnic enclave are at high risk of osteoporosis and to refer participants at high risk for follow-up care. DESIGN: Cross-sectional survey and osteoporosis screening, undertaken as a collaborative project by the Chinese American Service League and researchers at the University of Chicago. SETTING: Chicago's Chinatown. PARTICIPANTS: Four hundred sixty-nine immigrant Chinese American men and women aged 50 and older. MEASUREMENTS AND MAIN RESULTS: Chinese Americans in this urban setting are generally recent immigrants from south China with limited education and resources: mean age at immigration was 54, 56% had primary only or no education, and 57% reported "fair" or "poor" self rated health. Eighteen percent are uninsured and 55% receive Medicaid. Bone mineral density (BMD) of the calcaneus was estimated using quantitative ultrasound. Immigrant Chinese women in the study had lower average BMD than reference data for white women or U.S.-born Asian Americans. BMD for immigrant Chinese men in the study was similar to white men at ages 50 to 69, and lower at older ages. Low body mass index, low educational attainment and older age at immigration were all associated with lower BMD. CONCLUSIONS: Foreign-born Chinese Americans may be a high-risk group for osteoporosis. PMID- 12848834 TI - Encouraging Vietnamese-American women to obtain Pap tests through lay health worker outreach and media education. AB - BACKGROUND: Five times more Vietnamese-American women develop cervical cancer than white women. Few studies have examined whether community-based participatory research can effectively address Asian immigrants' health problems. This article reports the preliminary evaluation of 1 such project. METHODS: A coalition of 11 organizations in Santa Clara County, California worked with university researchers to design and simultaneously implement a media education (ME) campaign and a lay health worker outreach (LHWO) program to increase Vietnamese American women's cervical cancer awareness, knowledge, and screening. Two agencies each recruited 10 lay health workers (LHWs), who, in turn, each recruited 20 women who were then randomized into 2 groups: 10 to LHWO+ME (n = 200) and 10 to ME alone (n = 200). LHWs organized meetings with women to increase their knowledge and to motivate them to obtain Pap tests. Participants completed pre- and post-intervention questionnaires. RESULTS: At post-intervention, significantly more LHWO+ME women understood that human papillomavirus and smoking cause cervical cancer. The number of women who had obtained a Pap test increased significantly among women in both LHWO+ME and ME groups, but substantially more in the LHWO+ME group. Significantly more LHWO+ME women said they intended to have a Pap test. CONCLUSIONS: Media education campaigns can increase Vietnamese women's awareness of the importance of Pap tests, but lay health workers are more effective at encouraging women to actually obtain the tests. Lay health workers are effective because they use their cultural knowledge and social networks to create change. Researchers, community members, and community-based organizations can share expert knowledge and skills, and build one another's capacities. PMID- 12848835 TI - Internet access and empowerment: a community-based health initiative. AB - OBJECTIVE: To determine whether access to health information via in-home Internet technology can positively influence empowerment among residents of a low-income urban community. DESIGN: In-home Internet access and training were provided to volunteers, who, along with a comparison group, were interviewed prior to and 1 year after initiation of the program. Community-based participatory research methods were used to design and implement the intervention. SETTING: A 57-block area on the West Side of Chicago. PATIENTS/PARTICIPANTS: Twenty-five community residents completed all phases of the technology intervention. Thirty-five randomly selected neighbors of these residents served as the comparison group. INTERVENTIONS: Members of the intervention group received Internet access via WebTV, training, technical support, and access to a community specific health oriented web page during the course of the study. MEASUREMENTS AND MAIN RESULTS: Intervention group members were similar to comparison group members in terms of empowerment at baseline. After receiving Internet access and training, empowerment related to health decision-making improved significantly in the intervention group. Similar changes did not occur in the comparison group. Affinity for and appreciation of information technology also increased in the intervention group but not in the comparison group. As a result, differences in attitudes toward technology increased between the 2 groups over time. CONCLUSIONS: Using community-based participatory research methods, we found that Internet access to community-specific and general health information can lead to increased empowerment and appreciation of information technology. These benefits accrued among the intervention group but not among a random group of their neighbors. PMID- 12848836 TI - Trust, benefit, satisfaction, and burden: a randomized controlled trial to reduce cancer risk through African-American churches. AB - BACKGROUND: Community-based participatory research (CBPR) approaches that actively engage communities in a study are assumed to lead to relevant findings, trusting relationships, and greater satisfaction with the research process. OBJECTIVE: To examine community members' perceptions of trust, benefit, satisfaction, and burden associated with their participation. DESIGN, SETTING, AND PARTICIPANTS: A randomized controlled trial tested a cancer prevention intervention in members of African-American churches. Data were collected at baseline and 1-year follow-up. MEASUREMENTS: Subscales measured perception of trust in the research project and the project team, benefit from involvement with the project, satisfaction with the project and the team, and perception of burden associated with participation. MAIN RESULTS: Overall, we found high levels of trust, perceived benefit, and satisfaction, and low perceived burden among community members in Partnership to Reach African Americans to Increase Smart Eating. In bivariate analyses, participants in the intervention group reported more perceived benefit and trust (P <.05). Participants in smaller churches reported more benefit, satisfaction and trust, while participants from churches without recent health activities perceived greater benefit, greater satisfaction, and lower burden with the project and the team (P <.05). Participants whose pastors had less educational attainment noted higher benefit and satisfaction; those whose pastors were making personal lifestyle changes noted higher benefit and satisfaction, but also reported higher burden (P <.05). CONCLUSIONS: A randomized clinical trial designed with a CBPR approach was associated with high levels of trust and a perceived benefit of satisfaction with the research process. Understanding variations in responses to a research partnership will be helpful in guiding the design and implementation of future CBPR efforts. PMID- 12848837 TI - A community-centered approach to diabetes in East Harlem. AB - OBJECTIVE: Residents of East Harlem, an impoverished, non-white community in New York city (NYC), have up to 5 times the mortality and complication rates of diabetes compared with NYC residents overall. To determine potentially remediable problems underlying this condition, a community-based collaboration of health providers, community advocates, and researchers, surveyed East Harlem residents with diabetes to assess their knowledge, behaviors, barriers to care, and actions taken in response to barriers. DESIGN: Telephone interviews. SETTING: The 3 hospitals and 2 community clinics serving East Harlem. PARTICIPANTS: Nine hundred thirty-nine of the 1,423 persons (66%) with diabetes identified from these 5 healthcare sites with 2 or more ambulatory visits for diabetes during 1998 who lived in East Harlem. RESULTS: While most respondents (90%) said they know how to take their medicines, between 19% and 39% do not understand other aspects of their diabetes management. Many limit their diabetes care due to concerns about money (16% to 40%), and other barriers, such as language and transportation (19% to 22%). In multivariate analyses, Latinos (relative risk [RR] = 0.77; 95% confidence interval [95% CI] 0.63 to 0.91) and those who do not keep a diabetic diet due to concerns about money (RR = 0.85; 95% CI 0.70 to 0.99) had poorer health status. CONCLUSIONS: A community-based coalition was able to come together, identify areas of concern in diabetes care and assess the needs of adults with diabetes residing and obtaining care in East Harlem. The coalition found that even among those with access to care there remain significant financial barriers to good diabetes care, and a need to address and optimize how individuals with diabetes manage their disease. PMID- 12848838 TI - Religious involvement, social support, and health among African-American women on the east side of Detroit. AB - BACKGROUND: A significant body of research suggests that religious involvement is related to better mental and physical health. Religion or spirituality was identified as an important health protective factor by women participating in the East Side Village Health Worker Partnership (ESVHWP), a community-based participatory research initiative on Detroit's east side. However, relatively little research to date has examined the mechanisms through which religion may exert a positive effect on health. OBJECTIVE: The research presented here examines the direct effects of different forms of religious involvement on health, and the mediating effects of social support received in the church as a potential mechanism that may account for observed relationships between church attendance and health. DESIGN: This study involved a random sample household survey of 679 African-American women living on the east side of Detroit, conducted as part of the ESVHWP. MAIN RESULTS: Results of multivariate analyses show that respondents who pray less often report a greater number of depressive symptoms, and that faith, as an important source of strength in one's daily life, is positively associated with chronic conditions such as asthma or arthritis. Tests of the mediating effect of social support in the church indicated that social support received from church members mediates the positive relationship between church attendance and specific indicators of health. CONCLUSIONS: These findings are consistent with the hypothesis that one of the major ways religious involvement benefits health is through expanding an individual's social connections. The implications of these findings for research and practice are discussed. PMID- 12848839 TI - Community action against asthma: examining the partnership process of a community based participatory research project. AB - BACKGROUND: Community Action Against Asthma (CAAA) is a community-based participatory research (CBPR) project that assesses the effects of outdoor and indoor air quality on exacerbation of asthma in children, and tests household- and neighborhood-level interventions to reduce exposure to environmental asthma triggers. Representatives of community-based organizations, academia, an integrated health system, and the local health department work in partnership on CAAA's Steering Committee (SC) to design and implement the project. OBJECTIVE: To conduct a process evaluation of the CAAA community-academic partnership. DESIGN: In-depth interviews containing open-ended questions were conducted with SC members. Analysis included established methods for qualitative data, including focused coding and constant comparison methods. SETTING: Community setting in Detroit, Michigan. PARTICIPANTS: Twenty-three members of the CAAA SC. MEASUREMENTS: Common themes identified by SC members relating to the partnership's ability to achieve project goals and the successes and challenges facing the partnership itself. MAIN RESULTS: Identified partnership accomplishments included: successful implementation of a complex project, identification of children with previously undiagnosed asthma, and diverse participation and community influence in SC decisions. Challenges included ensuring all partners' influence in decision-making, the need to adjust to "a different way of doing things" in CBPR, constraints and costs of doing CBPR felt by all partners, ongoing need for communication and maintaining trust, and balancing the needs of science and the community through intervention. CONCLUSIONS: CBPR can enhance and facilitate basic research, but care must be given to trust issues, governance issues, organizational culture, and costs of participation for all organizations involved. PMID- 12848840 TI - Improving the nutritional resource environment for healthy living through community-based participatory research. AB - OBJECTIVES: To build health promotion capacity among community residents through a community-based participatory model, and to apply this model to study the nutritional environment of an urban area to better understand the role of such resources in residents' efforts to live a healthy life. DESIGN: A multiphase collaborative study that inventoried selected markets in targeted areas of high African-American concentration in comparison with markets in a contrasting wealthier area with fewer African Americans. SETTING: A community study set in the Los Angeles metropolitan area. PARTICIPANTS: African-American community organizations and community residents in the target areas. INTERVENTIONS: Two surveys of market inventories were conducted. The first was a single-sheet form profiling store conditions and the availability of a small selection of healthy foods. The second provided detailed information on whether the store offered fruit, vegetables, low-fat dairy products, dried goods and other items necessary for residents to consume a nutritious diet. RESULTS: The targeted areas were significantly less likely to have important items for living a healthier life. The variety and quality of fresh fruit and vegetable produce was significantly lower in the target areas. Such products as 1% milk, skim milk, low-fat and nonfat cheese, soy milk, tofu, whole grain pasta and breads, and low-fat meat and poultry items were significantly less available. CONCLUSIONS: Healthy food products were significantly less available in the target areas. The authors conclude from these results that the health disparities experienced by African American communities have origins that extend beyond the health delivery system and individual behaviors inasmuch as adherence to the healthy lifestyle associated with low chronic disease risk is more difficult in resource-poor neighborhoods than in resource-rich ones. PMID- 12848841 TI - Academic incentives for faculty participation in community-based participatory research. AB - Recognizing the need to overcome the obstacles of traditional university- and discipline-oriented research approaches, a variety of incentives to promote community-based participatory research (CBPR) are presented. Experiences of existing CBPR researchers are used in outlining how this methodological approach can appeal to faculty: the common ground shared by faculty and community leaders in challenging the status quo; opportunities to have an impact on local, regional, and national policy; and opening doors for new research and funding opportunities. Strategies for promoting CBPR in universities are provided in getting CBPR started, changing institutional practices currently inhibiting CBPR, and institutionalizing CBPR. Among the specific strategies are: development of faculty research networks; team approaches to CBPR; mentoring faculty and students; using existing national CBPR networks; modifying tenure and promotion guidelines; development of appropriate measures of CBPR scholarship; earmarking university resources to support CBPR; using Institutional Review Boards to promote CBPR; making CBPR-oriented faculty appointments; and creating CBPR centers. PMID- 12848842 TI - Integration of a participatory research strategy into a rural health survey. AB - The Glades Health Survey, a population-based survey of tuberculosis and HIV infection, provides a model for building community-research partnerships with local health departments in ethnically diverse communities. The survey was initiated without broad community participation; a year and a half of organizing established community leadership of the project. Essential factors in the success of the project included a shared objective, direct confrontation of fears about research, inclusion of all socioeconomic and racial/ethnic groups, and community participation in performing the research. These activities led to establishment of a community-based organization that received funding for HIV counseling and testing and HIV prevention case management. PMID- 12848843 TI - Moving from rhetoric to evidence-based action in health care. PMID- 12848844 TI - Community-based participatory research: opportunities, challenges, and the need for a common language. PMID- 12848845 TI - Murine dendritic cell development: difficulties associated with subset analysis. AB - Dendritic cells are bone marrow-derived professional antigen presenting cells that play major roles in both the induction of primary immune responses and tolerance. It has become clear that dendritic cells are a heterogeneous group of cells that vary in cell surface marker expression and function. Multiple dendritic cell subsets have now been defined in mouse lymphoid organs and peripheral tissues. A knowledge of the function and relationship between dendritic cell subsets will be essential for understanding the regulation of immune homeostasis, immune responses and tolerance. While an increasing number of dendritic cell progenitors are being identified, the pathways that connect them remain unclear. In addition, it is unclear whether the functional divisions reflect maturation status, subset specialization or functional plasticity in response to specific pathogen and environmental signals. This review summarizes the current knowledge about the function and lineage relationship of dendritic cell subsets. It also discusses some of the difficulties associated with dendritic cell subset analysis. PMID- 12848846 TI - Tryptophan and the immune response. AB - The immune system continuously modulates the balance between responsiveness to pathogens and tolerance to non-harmful antigens. The mechanisms that mediate tolerance are not well understood, but recent findings have implicated tryptophan catabolism through the kynurenine metabolic pathway as one of many mechanisms involved. The enzymes that break down tryptophan through this pathway are found in numerous cell types, including cells of the immune system. Some of these enzymes are induced by immune activation, including the rate limiting enzyme present in macrophages and dendritic cells, indoleamine 2,3-dioxygenase (IDO). It has recently been found that inhibition of IDO can result in the rejection of allogenic fetuses, suggesting that tryptophan breakdown is necessary for maintaining aspects of immune tolerance. Two theories have been proposed to explain how tryptophan catabolism facilitates tolerance. One theory posits that tryptophan breakdown suppresses T cell proliferation by dramatically reducing the supply of this critical amino acid. The other theory postulates that the downstream metabolites of tryptophan catabolism act to suppress certain immune cells, probably by pro-apoptotic mechanisms. Reconciling these disparate views is crucial to understanding immune-related tryptophan catabolism and the roles it plays in immune tolerance. In this review we examine the issue in detail, and offer additional insight provided by studies with antibodies to quinolinate, a tryptophan catabolite which is also necessary for nicotinamide adenine dinucleotide (NAD +) production. In addition to the immunomodulatory actions of tryptophan catabolites, we discuss the possible involvement of quinolinate as a means of replenishing NAD + in leucocytes, which is depleted by oxidative stress during an immune response. PMID- 12848847 TI - Cytotoxic T cells with reciprocal antigenic peptide presentation function are not generally resistant to mutual lysis. AB - Cytotoxic T cells normally express major histocompatibility complex class I molecules, to which their T cell antigen receptors are restricted. Therefore, a single cytotoxic T cell can not only act as a cytolytic effector cell, but also as an antigen-presenting cell for other cytotoxic T cells of the same or a different clone. In the present paper, we used a murine cytotoxic T cell clone, 10BK.1, recognizing the ovalbumin-derived peptide OVA257-264 in combination with H-2Kb to investigate the consequences of reciprocal antigen presentation by these cytotoxic T cells. These cells proliferate after incubation with the relevant peptide in the absence of added accessory cells, indicating reciprocal antigenic peptide presentation by the cytotoxic T cell. We found that reciprocal lysis of these cells was dependent on the time point of incubation with antigen. We did not observe reciprocal lysis of cytotoxic T cells used 30 days after the last restimulation with antigen. In contrast, 10BK.1 cells used two days after the last restimulation showed an increased capacity for reciprocal lysis. The lytic capacity decreased with time after restimulation. Reciprocal lysis of 10BK.1 cells depended on reciprocal peptide presentation by at least two 10BK.1 cells. Recognition of the antigenic peptide, together with class I molecules on the surface of classical syngeneic target cells did not induce lysis of freshly stimulated 10BK.1 cells, suggesting that reciprocal lysis was not just a consequence of re-activation of the cytotoxic T cells. Reciprocal destruction of freshly activated 10BK.1 cells proceeded independent of CD95/CD95 ligand. Despite an increased secretion of tumour necrosis factor-alpha by 10BK.1 cells on day 2 after antigen stimulation, compared with cells on day 30 after stimulation, tumour necrosis factor-alpha was not responsible for the reciprocal destruction of freshly stimulated 10BK.1 cells. Lysis of preactivated 10BK.1 cells was independent of autocrine interleukin-2 production by the cytotoxic T cells, but interleukin-2 was required for optimal priming of cytotoxic T cells for reciprocal lysis. PMID- 12848848 TI - mRNA of MUC2 is stimulated by IL-4, IL-13 or TNF-alpha through a mitogen activated protein kinase pathway in human colon cancer cells. AB - MUC2 mucin is a secretory glycoprotein which is produced from the intestinal goblet cells and is a major component of the intestinal epithelial mucus. The biological function of MUC2 mucin is considered to be the protection of intestinal epithelial surface, whereas the regulatory mechanism of MUC2 mucin production in immune response is not completely understood. We have studied the effects of cytokines, IL-4, IL-13 and TNF-alpha, on the regulation of MUC2 mRNA in the human colonic cancer cell lines, LS174T and HT29. The quantitative reverse transcription-polymerase chain reaction showed that single addition of IL-4, IL 13 and TNF-alpha to cell culture induced about two-fold increase of MUC2 mRNA level in LS174T cells. Interleukin-4 and IL-13 activated phosphorylation of mitogen-activated protein kinase in LS174T cells. A specific inhibitor of mitogen activated protein kinase pathway, U0126, totally inhibited the increase of MUC2 mRNA by IL-4 or IL-13 in those cells. Therefore, mitogen-activated protein activation of kinase is required for the increase of MUC2 mRNA by IL-4 or IL-13 in LS174T cells. In contrast to LS174T cells, only TNF-alpha increased MUC2 mRNA through a mitogen-activated protein kinase pathway in HT29 cells that express low levels of MUC2 mRNA. These findings sustain a novel phenomenon that MUC2 mRNA expression is differently controlled by IL-4, IL-13, or TNF-alpha in LS174T and HT29 cells, whereas the mitogen-activated protein kinase pathway plays a role in the MUC2 mRNA expression induced by those cytokines in both cell lines. PMID- 12848849 TI - Sustained linked stimulation via CD3 and CD4 is required for the IL-4-independent development of IL-4 synthesizing CD4+ T cells. AB - Previous work has shown that CD4 engagement can promote the development of interleukin-4-producing cells from naive CD4+ T cells activated with anti-CD3 antibody and interleukin-2 in the absence of other exogenous signals, including interleukin-4 itself. When CD44low CD4+ T cells were activated with immobilized anti-CD3 antibody and interleukin-2, they proliferated and produced interferon gamma but not interleukin-4. Co-immobilization of antibodies to CD3 and CD4 enhanced cell recoveries and induced interleukin-4 as well as interferon-gamma synthesis. Here we show that these effects of CD4 ligation were not observed when anti-CD4 antibody was replaced with another CD4 ligand, interleukin-16, or when the anti-CD3 and anti-CD4 antibodies were spatially separated by immobilization on different beads. Removal of the anti-CD4 antibodies within the first three days of stimulation also prevented the development of detectable interleukin-4 producing cells. The data suggest that interleukin-4-independent priming of interleukin-4-producing cells in this system requires sustained stimulation via both the T cell receptor and CD4 with close physical association between the ligands for these two receptors. PMID- 12848850 TI - Alveolar macrophages from subjects with chronic obstructive pulmonary disease are deficient in their ability to phagocytose apoptotic airway epithelial cells. AB - Chronic obstructive pulmonary disease is a highly prevalent, complex disease, usually caused by cigarette smoke. It causes serious morbidity and mortality and costs the global community billions of dollars per year. While chronic inflammation, extracellular matrix destruction and increased airway epithelial cell apoptosis are reported in chronic obstructive pulmonary disease, the understanding of the basic pathogenesis of the disease is limited and there are no effective treatments. We hypothesized that the accumulation of apoptotic airway epithelial cells chronic obstructive pulmonary disease in could be due to defective phagocytic clearance by alveolar macrophages. There have been no previous studies of the phagocytic capacity of alveolar macrophages in chronic obstructive pulmonary disease using physiologically relevant apoptotic airway epithelial cells as phagocytic targets. We developed a phagocytosis assay whereby cultured 16HBE airway epithelial cells were induced to apoptosis with ultraviolet radiation and stained with mitotracker green. Alveolar macrophages from bronchoalveolar lavage from eight control and six chronic obstructive pulmonary disease subjects were analysed following 1.5 h incubation with apoptotic airway epithelial cells, then staining with macrophage marker anti CD33. CD33+/mitotracker green + events (i.e., alveolar macrophages which had phagocytosed apoptotic airway epithelial cells) were analysed using flow cytometry. Phagocytosis of polystyrene microbeads was investigated in parallel. A significantly reduced proportion of alveolar macrophages from chronic obstructive pulmonary disease subjects ingested apoptotic airway epithelial cells compared with controls (11.6 +/- 4.1% for chronic obstructive pulmonary disease versus 25.6 +/- 9.2% for control group). Importantly, the deficiency was not observed using polystyrene beads, suggesting that the failure to resolve epithelial damage in chronic obstructive pulmonary disease may result, at least partially, from specific defects in phagocytic ability of alveolar macrophages to ingest apoptotic airway epithelial cells. PMID- 12848851 TI - Knockout B lymphoma cell lines as biochemical tools to explore multiple signalling pathways. AB - Studies on B lymphocyte signalling pathways using B lymphocytes from genetically modified mice have the disadvantages of primary cell polyclonality and finite life span. B lymphoma cell lines have been generated from mice with targeted mutations in the oct-2, OBF-1, vav-1 and btk genes, as a model system that lacks these limitations and possesses additional potential for experimental manipulation. To assess their utility, activation of the B cell receptor using anti- micro, the Toll-like receptor-4 using lipopolysaccharide and the interleukin-4 receptor were assessed in these cell lines. Differential tyrosine phosphorylation of intracellular proteins was measured in the wild-type controls compared to the corresponding mutant cell lines after B cell receptor stimulation. Intracellular calcium (Ca2+i) was mobilized in the control cell lines but not in the OBF-1 and Vav1-deficient cells, while Xid B cell lines (btk mutant) showed a reduced Ca2+ mobilization. Extracellular signal-regulated kinase 1/2 phosphorylation in response to anti- micro or lipopolysaccharide stimulation was significantly reduced in Vav1-deficient cells. Interleukin-4 stimulation of wild-type cells resulted in a 2-3-fold increase in Stat-6 phosphorylation. These results indicate that the cell lines mimic the biochemical responses of the corresponding primary B cells. They therefore represent a useful model system to investigate the regulation and roles of these and other gene products in B cell signal transduction and activation. PMID- 12848852 TI - Optimization of an ovine antibody-secreting cell assay for detection of antigen specific immunoglobulin production in peripheral blood leukocytes. AB - Enumeration of antibody-secreting cells in peripheral blood by enzyme-linked immunospot (ELISPOT) has been used in human studies to detect antigen-specific antibody production at mucosal tissue sites. An alternative assay for detecting and quantitating antigen-specific antibody responses involves culturing circulating peripheral blood antibody-secreting cells and quantitating specific antibody production in culture supernatant by ELISA. In the present study, antigen-specific peripheral blood lymphocytes were isolated from subcutaneously immunized sheep and the parameters for maximizing in vitro antibody production by in vivo-induced antibody-secreting cells optimized for this species. Maximum antibody-secreting cell responses were observed in peripheral blood collected four days after antigen challenge. The addition of lipopolysaccharide and antisheep immunoglobulin had no effect on in vitro antibody secretion by blood antibody-secreting cells, while the effects of pokeweed mitogen were highly variable. However, the combination of anti-Ig and recombinant ovine interleukin-6 to peripheral blood lymphocyte cultures was found to markedly and consistently enhance specific antibody production. In unstimulated cultures, the optimal peripheral blood lymphocyte concentration for generating the greatest antibody responses was 5.0 x 107 cells per mL, but in cultures stimulated with recombinant ovine interleukin-6/antisheep immunoglobulin, the optimal cell concentration was lowered to approximately 1.0 x 107 cells per mL. In vitro, peak immunoglobulin production was usually achieved by day one in unstimulated cultures. In recombinant ovine interleukin-6/antisheep immunoglobulin-stimulated cultures, antibody levels were similar to unstimulated cultures by day one, however, the levels continued to rise during incubation to reach a maximum between days four and five of incubation. This optimized antibody-secreting cell culture assay is amenable for increasing the sensitivity and reducing the cell numbers required for quantitating antigen-specific antibody induction in large-scale immunization trials in sheep and other large animal species. PMID- 12848853 TI - A novel CD44 antibody identifies an epitope that is aberrantly expressed on acute lymphoblastic leukaemia cells. AB - Previous studies have shown that the antibody 7H9D6 identifies CD44, a glycoprotein receptor for hyaluronic acid. 7H9D6 recognizes an epitope of CD44 that is not always present on CD44 molecules. The 7H9D6 antibody bound to the hyaluronic acid binding domain of CD44 and inhibited cell adhesion to immobilized hyaluronic acid. However, the expression of the 7H9D6 epitope was not sufficient for hyaluronic acid binding. Immunofluorescent staining with 7H9D6 revealed a punctate surface staining pattern, suggesting that CD44 molecules recognized by 7H9D6 are located in clusters on the cell surface. In contrast, other CD44 antibodies produced a uniform staining pattern. Early bone marrow B cells were negative for 7H9D6 but reactive with other CD44 monoclonal antibodies. In contrast, leukaemic cells from 65% of patients (28 of 43) with B lineage acute lymphoblastic leukaemia bound 7H9D6. Patients expressing the 7H9D6 epitope on their leukaemic cells had an increased risk of death (HR 3.5 95% CI 1.1-10.9, P = 0.029) and of disease relapse (HR 3.2 95% CI 1.2-8.5, P = 0.017) when corrected for white cell count. This antibody may be useful for the detection of residual disease in B lineage acute lymphoblastic leukaemia and as a prognostic indicator and for the study of CD44 function. PMID- 12848854 TI - Mucosal vascular addressin cell adhesion molecule-1 is expressed outside the endothelial lineage on fibroblasts and melanoma cells. AB - Mucosal vascular addressin cell adhesion molecule-1 (MAdCAM-1) is predominantly expressed on high endothelial venules in inflamed tissues where it assists with leucocyte extravasation. Here we report that MAdCAM-1 has the potential to be more widely expressed outside the endothelial cell lineage than previously appreciated. Thus, MAdCAM-1 RNA transcripts and cell-surface protein were expressed by NIH 3T3 fibroblasts following activation with tumour necrosis factor alpha (TNF-alpha), and by freshly isolated and cultured primary mouse splenic and tail fibroblasts in the absence of TNF-alpha stimulation. They were constitutively expressed by B16F10 melanoma cells, and expression was enhanced by cell activation with TNF-alpha. Mucosal vascular addressin cell adhesion molecule 1 was expressed on the apical surface of isolated cells, but became predominantly localized to cell junctions in confluent cell monolayers, suggesting it may play a role in the homotypic aggregation of cells. Tumour necrosis factor-alpha enhanced the expression of a firefly luciferase reporter directed by the MAdCAM-1 promoter in NIH 3T3 and B16F10 cells. A DNA fragment extending from nt -1727 to 673 was sufficient to confer cell-type selective expression. Mucosal vascular addressin cell adhesion molecule-1 expressed by NIH 3T3 cells was biologically active, as it supported the adhesion of TK-1 T cells in an alpha4beta7-dependent fashion. The expression of MAdCAM-1 by fibroblasts, and melanomas suggests MAdCAM 1 may play a role in regulating host responses in the periphery, leucocyte transmigration across nonendothelial boundaries, or the homotypic interactions of some malignant melanomas. PMID- 12848855 TI - Why do B cells produce CD40 ligand? AB - The CD40-CD40 ligand (CD40L) interaction is one of the most important receptor ligand interactions that occurs during a T dependent immune response. However, while CD40L is expressed on a range of cell types including activated T cells and B cells, dendritic cells granulocytes, macrophages and platelets, only CD40L on T cells is considered by most immunologists when planning experiments or analysing data. The current theory professes that T cells expressing CD40L can provide signals to B cells that induce proliferation, immunoglobulin class switching, antibody secretion, rescue from apoptosis at different times during the life of a B cell and also has a role in the development of germinal centres and the survival of memory B cells. However, the whole story is more complex than presently understood as human and mouse B cells express CD40L on their surface following activation and can release a soluble form of the ligand. This paper hypothesizes how CD40L on B cells may regulate antibody responses and the development of germinal centres. PMID- 12848859 TI - Visual quantitative estimation: semiquantitative wall motion scoring and determination of ejection fraction. AB - Ejection fraction (EF) is the most commonly used parameter of left ventricular (LV) systolic function and can be assessed by echocardiography. Quantitative echocardiography is time consuming and is as accurate as visual estimation, which has significant variability. We hypothesized that each echocardiographer has developed a mental set of guidelines that relate to how much individual segment shortening constitutes normal function or hypokinesis of varying extents. We determined the accuracy of applying these guidelines to an accepted technique of EF determination using a retrospective analysis of consecutive two-dimensional echocardiographic studies performed on patients who had radioventriculography (RVG) within 48 hours. Using a 12 segment model, we scored each segment at the base and mid-ventricular level based on segmental excursion and thickening. The apex was scored similarly but with 1/3 of the value based on a cylinder-cone model. EF was determined from the sum of segment scores and was estimated visually. We termed this approach visual quantitative estimation (VQE). We correlated the EF derived from VQE and visual estimation with RVG EF. In the training set, VQE demonstrated a strong correlation with RVG (r = 0.969), which was significantly greater than visual estimation (r = 0.896, P < 0.01). The limits of agreement for VQE (+12% to -7%) were similar to the limits of RVG agreement with contrast ventriculography (+10% to -11%) with similar intraobserver and interobserver variabilities. Similar correlation was noted in the prediction set between VQE and RVG EF (r = 0.967, P < 0.001). We conclude that VQE provides highly correlated estimates of EF with RVG. PMID- 12848860 TI - A method of detecting and quantifying severity of myocardial perfusion defects with intravenous ultrasound contrast and breath holding during stress echocardiography. AB - Although breath holding is commonly used to improve and maintain image quality during stress echocardiography, its effects on the qualitative and quantitative analysis of myocardial contrast enhancement (MCE) following intravenously injected microbubbles is unknown. The purpose of this study was to determine how breath holding affects MCE following either an intravenous bolus or continuous infusion of perfluorocarbon containing microbubbles. In 48 patients, intravenous Optison was given at peak dobutamine stress to assess myocardial perfusion. The degree of myocardial opacification was assessed immediately following a breath hold in inspiration (BH(ini)), at the end of a breath hold (BH(term)), and following expiration and a subsequent second breath hold (BH(reinsp)). Pulmonary venous time velocity integrals were recorded during these different phases as well. Eleven patients had quantitative coronary angiography. Mean duration of the breath hold was 7 +/- 1 seconds. Pulmonary venous return fell by 29% +/- 18% at BH(term) (P < 0.001). There was complete disappearance of MCE at BH(term) in 27 of 35 bolus injection patients at peak stress, and no return of MCE following flash destruction during breath holding in 11 of 13 patients receiving continuous infusions. BH(reinsp) resulted in a boluslike return of contrast, with a transient, bright MCE in 44 of 48 patients, and a time intensity plot that resembled a gamma variate function. Perfusion defects were visualized in 25 patients during BH(ini) and 28 patients during BH(reinsp). Coronary artery territory agreement between perfusion assessed during BH(ini) and BH(reinsp) and quantitative coronary angiography was 76% and 81%, respectively. PMID- 12848861 TI - Cyclic variation of myocardial integrated backscatter and myocardial wall thickness during percutaneous coronary angioplasty. AB - Cyclic variation of myocardial integrated backscatter (CVIBS) and change in myocardial wall thickness (WT%) were evaluated during percutaneous transluminal coronary angioplasty (PTCA). Fourteen patients who underwent PTCA of the proximal left anterior descending (LAD) coronary artery were included in the study. PTCA was performed by inflating the balloon at the site of the LAD lesion for 1 minute. CVIBS was measured at three episodes during PTCA in the parasternal short axis view: before the inflation, at the end of 1-minute inflation, and at the fifth-minute after deflation of the balloon. Three regions of interest were used to evaluate the three-vessel territories: mid-anteroseptal area for LAD, mid posterolateral area for circumflex artery, and mid-inferior area for right coronary artery. The WT% was calculated in each area. In the LAD territory, CVIBS measured at the end of 1-minute inflation was lower than the values obtained before PTCA, 5.2 +/- 1.0 decibel (dB) versus 3.7 +/- 0.7 dB (P < 0.01). CVIBS magnitudes increased at the fifth-minute after the deflation back up even to higher levels than pre-PTCA values,6.1 +/- 1.0 dB versus 5.2 +/- 1.0 dB (P < 0.01). The WT% values decreased during balloon inflation but did not recover to the pre-inflation values measured at fifth-minute after deflation. In other sites, there was no change in either CVIBS or WT% values at any time studied. The observed increase in CVIBS may be an indicator of restoration of blood flow to ischemic myocardium. PMID- 12848862 TI - Culture-negative suppurative endocarditis causing severe mitral valve obstruction: complementary use of transesophageal and transthoracic echocardiography. AB - Infective endocarditis causes a myriad number of serious complications. Mitral valve obstruction is a rare complication. We report a 48-year-old Asian female who presented with two-week duration of fever and rapidly developed acute pulmonary edema and cardiogenic shock. Sequential transthoracic and transesophageal echocardiography revealed a rapidly growing vegetation on the anterior mitral leaflet with severe stenosis of the valve. All the blood cultures were negative. The patient underwent a successful mitral valve replacement. A review of 21 previously reported cases of mitral valve obstruction from endocarditis demonstrates the poor prognosis of this entity and supports early surgery. PMID- 12848863 TI - Conservative surgical treatment of anterior mitral valve aneurysm secondary to aortic valve endocarditis. PMID- 12848864 TI - Undefined hypercoaguable state associated with massive right ventricular thrombus and embolism in a previously healthy 17-year-old male. AB - A right ventricular thrombus (RVT) is an unusual finding on echocardiography. We describe a healthy young male patient who developed RVT with subsequent pulmonary embolism (PE), the etiology of which remains uncertain. PMID- 12848865 TI - Normal lymph nodes mimicking a mediastinal mass during transesophageal echocardiography. AB - We report a transesophageal echocardiographic (TEE) study in an adult patient with ischemic stroke, in whom a nodular mass lesion consistent with tumor or mediastinal lymphadenopathy appeared to be interposed between the esophageal probe and the proximal descending aorta/aortic arch. Computed tomography (CT) scan of the chest revealed no pathologic lesions, only the presence of normal mediastinal lymph nodes and a rightward displaced esophagus. This unusual, but normal rightward position of the esophagus appears to have facilitated the visualization of these normal mediastinal lymph nodes on TEE. CT scan of the chest helped clarify the benign nature of these lymph nodes, which mimicked a mediastinal mass on TEE. PMID- 12848867 TI - The hand-carried ultrasound revolution. PMID- 12848866 TI - Three-dimensional transesophageal echocardiographic demonstration of intraatrial baffle obstruction. AB - We report an adult patient with transposition of the great arteries status post Mustard procedure in whom three-dimensional transesophageal echocardiography demonstrated intraatrial baffle obstruction. The baffle could be visualized in both long-axis and "en face" short-axis views. PMID- 12848868 TI - Hand carried echocardiography in the critical care setting. AB - Miniaturized echocardiography units known as hand carried ultrasound (HCU) now exist with features similar to those used for standard echocardiography. The small size and low cost of these units may lead to increased availability of echocardiography to be performed by all physicians taking care of critically ill patients. Use of HCU by critical care physicians may allow for improved bedside diagnosis with improved accuracy over physical examination. Studies comparing HCU with standard echocardiography in the critical care setting have reported that HCU is limited by decreased image quality and that it may miss important diagnoses even when used by experienced sonographers and echocardiographers. Despite its limitations, however, HCU can often answer important clinical questions in the critically ill. This review explores the current literature on the use of HCU in the critical care setting, discusses the limitations of HCU, and examines the costs of implementing this new technology. PMID- 12848869 TI - Utility of hand-carried ultrasound for consultative cardiology. AB - Although the stethoscope has been an important part of the bedside cardiac diagnostic examination for generations of physicians, this clinical tool has been relatively unchanged in over 150 years. Echocardiography is established as an essential diagnostic imaging method for patients with known or suspected cardiovascular diseases. However, routine echocardiography systems are large and heavy, and although they are portable, they remain inconvenient for bedside patient rounds. Technologic advances have resulted in miniaturization of electronic components and small, lightweight ultrasound systems have been recently introduced. These hand-carried units offer clinically acceptable two dimensional image quality for rapid "quick-look" bedside diagnostics, in particular focusing on global and regional left ventricular function and presence or absence of pericardial effusion. This article proposes a general approach to the rapid hand-carried ultrasound cardiac exam as an extension of the physical examination. It details case examples and reviews the initial clinical experience of hand-carried ultrasound on cardiac consultation rounds. Hand-carried ultrasound has promise to have an immediate impact on bedside patient management though expediting and facilitating the delivery of medical care. PMID- 12848870 TI - Can hand-carried ultrasound devices be extended for use by the noncardiology medical community? AB - Echocardiography (echo) is a powerful, noninvasive, inexpensive diagnostic imaging technique that provides important information in a variety of cardiovascular diseases. Echo provides rapid information regarding ventricular and valvular function in the clinical management of patients. Smaller, relatively inexpensive hand-carried cardiac ultrasound (HCU) devices have become commercially available, which can be used for diagnostic cardiac imaging. Because of their relative ease of use, portability, and affordable cost, these new hand held systems have made point-of-care (bedside) echocardiography available to all medical personnel. The rate-limiting step to the widespread use of this technology is the lack of personnel with echo training at the immediate contact point with patients. Although extensive training and experience are needed to acquire and interpret a complete echo, training medical personnel to perform and interpret a limited echo (defined as a brief, diagnosis focused exam) may fully exploit the potential of echo as a point-of-care diagnostic tool and may be accomplished in a short period of time. Presently there are guidelines for independent competency in echocardiography and HCU echo established by several professional organizations and as a result of these rigorous guidelines, other noncardiology medical professionals who could practically derive the greatest benefit are discouraged and virtually precluded from utilizing echo during the initial encounter with the patient. However, there is now a growing body of literature in a diverse group of noncardiology medical personnel that demonstrates that it is possible to quickly and effectively train them to perform and interpret limited echocardiograms. Medical students, medical residents, cardiology fellows with limited experience, emergency department physicians, and surgical intensive care unit staff have all been evaluated after only brief, focused training periods, and investigators found that HCU echo provided important new information, changed therapeutic management, and was vastly superior to the physical exam alone with an acceptable overall level of accuracy. The contribution of echocardiography to the field of cardiovascular disease since its invention has been significant and the newer compact, portable, ultrasound systems have the potential to revolutionize the utilization and availability of echocardiography. To maximize integration of echo into medical practice, physicians and physician extenders could be trained to perform and interpret limited echo to complement their clinical examination and improve their diagnostic skills. The challenge is to provide practical training programs to assure competency in performing point of care echocardiograms. PMID- 12848871 TI - The hand-carried echocardiographic device as an aid to the physical examination. AB - Physical examination skills have been declining over the past several decades while technology has made diagnostic testing increasingly sophisticated. For patients with cardiovascular disease, the best approach to bedside diagnosis would be one that combines both physical examination and ready access to technology at the time of the patient encounter. Most cardiac testing is not performed at the bedside due to equipment size and time limitations for these tests. Small hand-carried echocardiographic devices are now available for rapid bedside examination. These devices compare well to full-featured systems when used in cardiology outpatient settings and in hospitalized patients who are not critically ill. Compared with physical examination by board certified cardiologists, these devices decrease diagnostic error. Early use of hand-carried echocardiographic devices after physical examination has been demonstrated to impact patient triage and treatment as well as uncover otherwise undetected cardiac disease. The degree of training required for responsible use of these devices is as yet unclear. However, organized training sessions have resulted in modest agreement with standard echocardiography and point-of-care echocardiography performed by expert echocardiographers. It is conceivable that the hand-carried echocardiographic devices will be used in medical school curriculum to enhance medical student education in the future. PMID- 12848872 TI - Accuracy of hand-carried ultrasound. AB - Hand-carried ultrasound introduces a new group of devices, operators and usage patterns to echocardiography. This may have significant impact on the accuracy of the findings obtained with hand-carried ultrasound. Although reasonable agreement can be obtained with standard echocardiography in certain circumstances, limitations in imaging modes, device image quality, operator experience, and study completeness may significantly limit the diagnostic accuracy of hand carried ultrasound. Despite this, hand-carried ultrasound has the potential to improve significantly upon the data obtained by physical examination. PMID- 12848873 TI - A hand-carried personal ultrasound device for rapid evaluation of left ventricular function: use after limited echo training. AB - A hand-carried personal ultrasound device (HCPUD) may be used for rapid cardiac screening by physicians with limited echo training. Our objective was to determine the accuracy of rapid HCPUD evaluation of left ventricular (LV) size and function when used by a Cardiology Fellow. Forty-five patients underwent an HCPUD exam using a 2.4-kg device with a 2- to 4-MHz curved transducer and color power Doppler (SonoSite). The results were compared with sonographer-performed and echocardiographer-interpreted exams using conventional equipment. The HCPUD exam lasted 6 +/- 2 minutes. There was 100% agreement between HCPUD and conventional echo on qualitative assessment of LV systolic function. Comparing the HCPUD and conventional linear measurements of left ventricular end-diastolic dimension (LVEDD) and of interventricular septal (IVS) thickness: LVEDD is HCPUD = 0.94 conventional -0.2,r = 0.82, P < 0.0001; IVS is HCPUD= 0.59 conventional+0.6, r = 0.69, P < 0.0001. Thus, an HCPUD can effectively be used after limited training to rapidly screen for qualitative abnormalities of LV systolic function. Quantitative measurements of smaller structures with the HCPUD are more challenging. PMID- 12848874 TI - Automated quantification of left ventricular function by the automated contour tracking method. AB - The automated contour tracking (ACT) method has been developed for the automated measurement of area volume using the energy minimization method without tracing a region of interest. The purpose of this study was to compare the ACT method and left ventriculography (LVG) for the measurement of left ventricular (LV) function in the clinical setting. An apical four-chamber view was visualized by two dimensional echocardiography and recorded for off-line analysis in 14 patients with high-quality images who underwent LVG. The ACT method automatically traces the endocardial border from the recorded images and calculates LV volumes (end diastole and end-systole) and ejection fraction (EF). Both ACT and LVG were compared by linear regression analysis for the measurement of EF. EF determined by the ACT method agreed well with that by LVG (r = 0.96, y = 0.94x + 4.6, standard error of the estimate = 3.9%). The mean difference between the ACT and LVG was -1.4%+/- 7.3%. In conclusion, the ACT method is reliable for noninvasive estimation of EF in high-quality images. This suggests that this new technique may be useful in the automated quantification of LV function. PMID- 12848875 TI - Influence of perindopril on left ventricular global performance during the early phase of inferior acute myocardial infarction: assessment by Tei index. AB - The beneficial effect of angiotensin-converting enzyme inhibitors (ACE inhibitors) on left ventricular (LV) function in patients with acute myocardial infarction (AMI) is widely known. However, controversy exists about their efficacy on patients with small infarcts and preserved LV systolic function. The aim of the present study was to detect the influence of the ACE-I perindopril on the global LV performance in patients with pure inferior AMI (AMI-I) using a Doppler-derived index (DI) that combines systolic and diastolic time intervals (Tei index). Our study included 40 patients with first AMI-I, mean age 60 years +/- 9.06 years (SD) and 24 age- and gender-matched normal patients who constituted the control group (COG). Patients were randomized into two groups to receive the conventional treatment of AMI-I (GCT) or the above therapy plus P (GP). Complete Doppler echocardiography (systolic and diastolic parameters), DI, and systolic blood pressure (SBP) were measured on the 8.07 +/- 1.16(SD) post infarct day. The same examination was performed to COG. The DI was significantly lower in healthy patients(0.45 +/- 0.23)compared with the value in patients of either GP(0.56 +/- 0.03; P = 0.023)or GCT(0.78 +/- 0.05; P = 0.000). Moreover DI was higher in patients of GCT compared with that of GP(P = 0.000). In addition, perindopril administration decreased isovolumic relaxation time(IRT; 120.00 +/- 4.23 vs. 139.00 +/- 6.74; P = 0.006)and increased significantly ejection time (ET;274.25 +/- 7.35 vs. 253.50 +/- 7.68; P = 0.042). SBP in patients of GP was similar to that of GCT(120.5 +/- 2.85 mmHg vs. 112.5 +/- 3.49 mmHg; P = NS). CONCLUSIONS: Global LV function (DI) is impaired in patients with AMI-I. Administration of perindopril has a favorable impact on LV performance in patients with AMI-I, achieved through improvement of the diastolic function (IRT), which indirectly improves LV systolic function (ET, DI). This beneficial influence of perindopril is the result of the direct tissue effect of the drug and not its hemodynamic action. PMID- 12848876 TI - Right ventricular myocardial adaptation to different training protocols in top level athletes. AB - OBJECTIVE: The aim of this study was to analyze right ventricular (RV) myocardial function in competitive athletes with left ventricular (LV) hypertrophy induced by either endurance or strength training. METHODS: Standard Doppler echo, maximal electrocardiogram (ECG) ergometric test, and pulsed tissue Doppler (TD) of LV mitral annulus and of RV tricuspid annulus were performed in 32 competitive endurance athletes (long-distance swimmers; ATE) and in 26 strength-trained athletes (short-distance swimmers; ATS), all males. By use of TD, the following parameters of myocardial function were assessed: systolic peak velocities (Sm), precontraction time, contraction time, early (Em) and late (Am) diastolic velocities, Em/Am ratio, and relaxation time. RESULTS: The two groups were comparable for age, but ATS at rest exhibited higher heart rate, systolic blood pressure, and body surface area. LV mass index did not significantly differ between the two groups. However, ATS characterized increased wall thickness and relative wall thickness, whereas LV stroke volume and both LV and RV end diastolic diameters were greater in ATE. All transmitral and transtricuspid Doppler indexes were higher in ATE, with increased E/A ratios. TD analysis demonstrated in ATE higher Em and Em/Am ratio as well as longer relaxation time both at mitral and at tricuspid annulus level. In the overall population, distinct multiple linear regression models evidenced independent positive association between RV peak Em velocity and LV end-diastolic diameter (P < 0.001) as well as independent direct correlation of the same RV peak Em velocity with both LV stroke volume and maximal workload achieved by bicycle ergometer (both P < 0.001). CONCLUSIONS: Right ventricular early diastolic myocardial function is positively influenced by preload increase in athletes, and represents an independent determinant of cardiac performance during physical effort. Therefore, pulsed TD may be taken into account to distinguish different cardiac adaptation to either endurance or strength sport activities, and eventually to quantify RV adaptation degree to long-term training. PMID- 12848877 TI - Integrated backscatter during harmonic and fundamental frequency imaging--effect of depth, mechanical index, and tissue anisotropy: implications for myocardial tissue characterization. AB - OBJECTIVE: To explore the potential advantages of tissue harmonic imaging (THI) versus fundamental frequency imaging (FFI) when applied to tissue characterization. METHODS: A Philips Medical Systems Sonos 5500 echocardiograph equipped with a broadband transducer (S4) and an on-line quantitative analysis software package (Acoustic Densitometry) was used for imaging. The effect of mechanical index (MI), imaging depth, and anisotropy on relative backscatter amplitude was evaluated. RESULTS: This study demonstrated that imaging with tissue harmonics generated relatively greater backscatter values at clinically relevant imaging depths and instrument settings referenced to FFI. This effect was dependent on MI setting. A direct relationship between backscatter amplitude and MI was demonstrated. Additionally, tissue anisotropy had similar effects on integrated backscatter amplitude during both THI and FFI. However, relative backscatter values at each fiber orientation are greater during THI at similar instrument settings when referenced to FFI. CONCLUSION: Tissue harmonic imaging may offer advantages over FFI for myocardial tissue characterization. PMID- 12848878 TI - Detection of a pseudonormal mitral inflow pattern: an echocardiographic and tissue Doppler study. AB - OBJECTIVE: The aim of this study was to assess the ability of several echocardiographic and tissue Doppler imaging (TDI) derived parameters to improve the noninvasive diagnosis of a pseudonormal mitral inflow pattern. METHODS: Ninety-eight consecutive patients with age-related normal transmitral Doppler profile underwent echocardiography including TDI and measurement of left ventricular end-diastolic pressure (LVEDP) using fluid-filled catheters. Peak transmitral velocities were determined at rest (E, A) and during the strain phase of a Valsalva maneuver. The difference in duration between the pulmonary venous retrograde velocity and the transmitral A-velocity (PVR-A) was calculated from pulsed Doppler recordings. Propagation velocity of the early mitral inflow (VP) was determined by color M-mode. Early diastolic peak mitral annulus velocities (E') and the early diastolic transmyocardial velocity gradient of the posterior basal wall (MVG) were obtained by TDI. RESULTS: Fifty-two patients presented with normal diastolic function (group I: LVEDP9.5 +/- 3 mm Hg, E/A1.1 +/- 0.19), while pseudonormalization, defined as LVEDP 15 mm Hg and E/A > 0.9, was found in 46 patients (group II: LVEDP23 +/- 7 mm Hg, E/A1.43 +/- 0.83). The coefficient of linear correlation (r) and the area under ROC - curve (AUC) to predict LVEDP values 15 mm Hg were maximal for the index PVR-A (AUC = 0.92, r = 0.77), followed byE/E' (AUC = 0.80, r = 0.46), MVG (AUC = 0.65, r = 0.33) and E/VP (AUC = 0.69, r = 0.30), P < 0.01, whereas the decrease in E/A ratio during Valsalva maneuver failed to reach significance. Similar results were observed when echocardiographic parameters were used to estimate the left ventricular diastolic pressure before atrial contraction. CONCLUSIONS: PVR-A enabled the most accurate estimation of LVEDP. TDI-derived indices E/E' and MVG are also reliable alternatives superior to the classical Valsalva maneuver to detect a pseudonormal transmitral Doppler profile. PMID- 12848879 TI - Anatomical M-mode: an old-new technique. AB - The M-mode (motion mode) technique has improved the diagnostic possibilities of echocardiography due to its high temporal resolution. The use of a two dimensional (2D) image as a basis for M-mode analysis at a defined line, independent of the transducer orientation, namely the anatomic M-mode (AMM), has been proposed from the beginning of 2D echocardiography. For several years, however, this could not be accomplished due to several reasons including the limited digital memory, the relatively rough pixels of 2D images, and the low temporal resolution of the screen. The AMM has been improved by the "fully digital" machines. These are able to provide a series of digital data (direction, position, and timing) relative to any single echo received from any point of the tissue. AMM analysis, thus, can be performed in any direction, as a "normal" monodimensional echocardiogram. With respect to traditional M-mode, AMM permits a more detailed analysis of cardiac chambers diameters obtained by linear measurements, regional wall motion of the left ventricle (both at rest and during stress), and location of accessory pathways. In particular, the assessment of left ventricular regional wall motion represents the most important goal of this new technique, which results in marked reduction or even elimination of the limitations due to the subjective character of wall motion evaluation with 2D echocardiography. PMID- 12848880 TI - Mobile components associated with rapidly developing mitral annulus calcification in patients with chronic renal failure: review of mobile elements associated with mitral annulus calcification. AB - Mitral annulus calcification may directly cause cerebrovascular accidents by serving as a source of calcific or thrombotic emboli. This hypothesis has been supported by recent reports of mobile components associated with mitral annulus calcification detected by echocardiography. Cardiovascular calcifications, including mitral annulus calcification, are common in end-stage renal disease and may develop and progress over a short period of time. We report two patients with mobile components associated with mitral annulus calcification and end-stage renal disease. Serial echocardiography documented that the mitral annulus calcification in these two patients had developed rapidly. Follow-up echocardiography in one patient demonstrated resolution of the mobile elements. PMID- 12848881 TI - Eosinophilic heart: marked left ventricular wall thickening and myocardial dysfunction improving with corticosteroid therapy. AB - We report a case of a 34-year-old male with acute severe heart failure associated with marked concentric left ventricular wall thickening and biopsy evidence of eosinophilic myocardial infiltrate. This appears to be an unusual description of this degree of concentric myocardial thickening in eosinophilic myocarditis coupled with Doppler tissue echocardiography. Following high-dose corticosteroid treatment, wall thickness, systolic and diastolic left ventricular function normalized and the patient experienced a dramatic clinical improvement. PMID- 12848882 TI - Pulmonary artery dissection: echocardiographic findings and diagnosis. AB - Pulmonary artery dissection is a rare but life-threatening event, predisposing to sudden cardiac death or cardiogenic shock. It is often associated with underlying congenital disorders predisposing to pulmonary hypertension. Rarely, it is diagnosed by echocardiography or other image techniques. We present a case report of a pulmonary artery dissection, diagnosed primarily by echocardiography. The patient died soon after refusing any interventional approach after a short period under medication. PMID- 12848883 TI - Noncompaction of the ventricular myocardium: report of two cases with bicuspid aortic valve demonstrating poor prognosis and with prominent right ventricular involvement. AB - Noncompaction of the ventricular myocardium is a rare, unclassified cardiomyopathy due to an arrest of myocardial morphogenesis. The characteristic echocardiographic findings consist of multiple, prominent myocardial trabeculations and deep intertrabecular spaces communicating with the left ventricular (LV) cavity. The disease typically involves the LV myocardium, but right ventricular (RV) involvement is not uncommon. The clinical manifestations include heart failure (HF) signs, ventricular arrhythmias and cardioembolic events. Noncompacted myocardium may occur as an isolated cardiac lesion, as well as it can be in association with congenital anomalies. We describe two illustrative cases of noncompaction of the ventricular myocardium, a 19-year-old male with bicuspid aortic valve and progressive worsening of HF, and a 61-year old male with marked RV involvement in addition to LV apical involvement, both with the typical clinical and echocardiographic features of the disease. PMID- 12848884 TI - Coincidence of sporadic left atrial myxoma and benign Schwannoma of the foot erroneously diagnosed as Carney complex. PMID- 12848885 TI - Metastatic hepatocellular carcinoma into the right atrium and ventricle: echocardiographic diagnosis and follow-up. PMID- 12848886 TI - Transcatheter closure of atrial septal defect under combined transesophageal and intracardiac echocardiography. PMID- 12848887 TI - Contrast transesophageal echocardiographic detection of a pulmonary arteriovenous malformation draining into left lower pulmonary vein. AB - We report the identification of a pulmonary arteriovenous malformation draining into the left lower pulmonary vein by contrast two-dimensional transesophageal echocardiography in an adult with no evidence of hereditary hemorrhagic telangiectasia. To our knowledge, this has not been reported previously. This study also emphasizes the importance of transesophageal echocardiographic examination of the left lower pulmonary vein in the detection of a pulmonary arteriovenous malformation. PMID- 12848888 TI - Transesophageal echocardiographic identification of bilateral vertebral artery ostial stenosis. AB - We describe the usefulness of transesophageal echocardiography in correctly diagnosing severe ostial stenosis of both the left and right vertebral arteries in an adult patient. Although transesophageal echocardiography has been previously used to identify left vertebral artery stenosis, to our knowledge this study represents the first demonstration of its usefulness in detecting bilateral vertebral artery ostial stenosis. PMID- 12848892 TI - Distal hinge of plasminogen activator inhibitor-1 involves its latency transition and specificities toward serine proteases. AB - BACKGROUND: The plasminogen activator inhibitor-1 (PAI-1) spontaneously converts from an inhibitory into a latent form. Specificity of PAI-1 is mainly determined by its reactive site (Arg346-Met347), which interacts with serine residue of tissue-type plasminogen activator (tPA) with concomitant formation of SDS-stable complex. Other sites may also play roles in determining the specificity of PAI-1 toward serine proteases. RESULTS: To understand more about the role of distal hinge for PAI-1 specificities towards serine proteases and for its conformational transition, wild type PAI-1 and its mutants were expressed in baculovirus system. WtPAI-1 was found to be about 12 fold more active than the fibrosarcoma PAI-1. Single site mutants within the Asp355-Arg356-Pro357 segment of PAI-1 yield guanidine activatable inhibitors (a) that can still form SDS stable complexes with tPA and urokinase plasminogen activator (uPA), and (b) that have inhibition rate constants towards plasminogen activators which resemble those of the fibrosarcoma inhibitor. More importantly, latency conversion rate of these mutants was found to be approximately 3-4 fold faster than that of wtPAI-1. We also tested if Glu351 is important for serine protease specificity. The functional stability of wtPAI-1, Glu351Ala, Glu351Arg was about 18 +/- 5, 90 +/- 8 and 14 +/- 3 minutes, respectively, which correlated well with both their corresponding specific activities (84 +/- 15 U/ug, 112 +/- 18 U/ug and 68 +/- 9 U/ug, respectively) and amount of SDS-stable complex formed with tPA after denatured by Guanidine-HCl and dialyzed against 50 mM sodium acetate at 4 degrees C. The second-order rate constants of inhibition for uPA, plasmin and thrombin by Glu351Ala and Glu351Arg were increased about 2-10 folds compared to wtPAI-1, but there was no change for tPA. CONCLUSION: The Asp355-Pro357 segment and Glu351 in distal hinge are involved in maintaining the inhibitory conformation of PAI-1. Glu351 is a specificity determinant of PAI-1 toward uPA, plasmin and thrombin, but not for tPA. PMID- 12848893 TI - Ovarian follicular activity and hormonal profile during estrous cycle in cows: the development of 2 versus 3 waves. AB - Most estrous cycles in cows consist of 2 or 3 waves of follicular activity. Waves of ovarian follicular development comprise the growth of dominant follicles some of which become ovulatory and the others are anovulatory. Ovarian follicular activity in cows during estrous cycle was studied with a special reference to follicular waves and the circulating concentrations of estradiol and progesterone. Transrectal ultrasound examination was carried out during 14 interovulatory intervals in 7 cows. Ovarian follicular activity was recorded together with assessment of serum estradiol and progesterone concentrations. Three-wave versus two-wave interovulatory intervals was observed in 71.4% of cows. The 3-wave interovulatory intervals differed from 2-wave intervals in: 1) earlier emergence of the dominant follicles, 2) longer in length, and 3) shorter interval from emergence to ovulation. There was a progressive increase in follicular size and estradiol production during growth phase of each wave. A drop in estradiol concentration was observed during the static phase of dominant anovulatory follicles. The size of the ovulatory follicle was always greater and produced higher estradiol compared with the anovulatory follicle. In conclusion, there was a predominance of 3-wave follicular activity that was associated with an increase in length of interovulatory intervals. A dominant anovulatory follicle during its static phase may initiate the emergence of a subsequent wave. Follicular size and estradiol concentration may have an important role in controlling follicular development and in determining whether an estrous cycle will have 2 or 3-waves. PMID- 12848894 TI - Epidermal Growth Factor Receptor: A Novel Target for Anticancer Treatment. PMID- 12848895 TI - A brief guide to the selection of quality of life instrument. AB - There are numerous quality of life (QOL) scales. Because QOL experts are often partial to their own scales, researchers need to be able to select scales for themselves. Scales best suited for longitudinal purposes (clinical trials and audit) have different properties to those suited for cross-sectional studies (population and correlational studies and clinical use). The reason and logic of these differences is explained. For longitudinal use, researchers need to consider the relationship between item set, population and treatment; scales can be short, floor and ceiling effects must be avoided, and there should be extended response options. For cross-sectional use scales should have a wide range of items, should be longer, and there are no adverse floor and ceiling effects, and response options can be simpler to allow a larger set of items. PMID- 12848896 TI - Potential role of the interaction between equine estrogens, low-density lipoprotein (LDL) and high-density lipoprotein (HDL) in the prevention of coronary heart and neurodegenerative diseases in postmenopausal women. AB - BACKGROUND: An inverse relationship between the level of high-density lipoprotein (HDL) and coronary heart disease (CHD) has been reported. In contrast, oxidized HDL (oHDL) has been shown to induce neuronal death and may play an important role in the pathogenesis of CHD. In the present study we have investigated a: the effect of various equine estrogens on HDL oxidation, b: the inhibition of LDL oxidation by HDL and c: the effect of these estrogens on LDL oxidation in the presence of HDL. RESULTS: All 11 equine estrogens tested protected the HDL from oxidation in a concentration dependant manner. Equilenin, 17beta dihydroequilenin, and 17alpha-dihydroequilenin (Delta6-8-estrogens) were found to be the most potent inhibitors of HDL oxidation. Some of the novel ring B unsaturated estrogens were 2.5 to 4 times more potent inhibitors of HDL oxidation than 17beta-estradiol. HDL was found to delay LDL oxidation. The protection of LDL oxidation by HDL is enhanced by the addition of estrogen, with equilenin being again more potent than 17beta-estradiol. CONCLUSIONS: Equine estrogens can differentially inhibit the oxidation of HDL with the Delta6-8-estrogens being the most potent antioxidants. The ability of estrogens to enhance HDL's antioxidant activity is to our knowledge the first report of an interaction of estrogen with HDL that results in the delay or inhibition of LDL oxidation. This may be another mechanism by which estrogens may reduce the risk of CHD and neurodegenerative diseases in healthy and younger postmenopausal women. PMID- 12848897 TI - Programmed cell death in trypanosomatids: is it an altruistic mechanism for survival of the fittest? AB - The protozoan parasites Leishmania, Trypanosoma cruzi and Trypanosoma brucei show multiple features consistent with a form of programmed cell death (PCD). Despite some similarities with apoptosis of mammalian cells, PCD in trypanosomatid protozoans appears to be significantly different. In these unicellular organisms, PCD could represent an altruistic mechanism for the selection of cells, from the parasite population, that are fit to be transmitted to the next host. Alternatively, PCD could help in controlling the population of parasites in the host, thereby increasing host survival and favoring parasite transmission, as proposed by Seed and Wenk. Therefore, PCD in trypanosomatid parasites may represent a pathway involved both in survival and propagation of the species. PMID- 12848898 TI - The acute neurotoxicity of mefloquine may be mediated through a disruption of calcium homeostasis and ER function in vitro. AB - BACKGROUND: There is no established biochemical basis for the neurotoxicity of mefloquine. We investigated the possibility that the acute in vitro neurotoxicity of mefloquine might be mediated through a disruptive effect of the drug on endoplasmic reticulum (ER) calcium homeostasis. METHODS: Laser scanning confocal microscopy was employed to monitor real-time changes in basal intracellular calcium concentrations in embryonic rat neurons in response to mefloquine and thapsigargin (a known inhibitor of the ER calcium pump) in the presence and absence of external calcium. Changes in the transcriptional regulation of known ER stress response genes in neurons by mefloquine were investigated using Affymetrix arrays. The MTT assay was employed to measure the acute neurotoxicity of mefloquine and its antagonisation by thapsigargin. RESULTS: At physiologically relevant concentrations mefloquine was found to mobilize neuronal ER calcium stores and antagonize the pharmacological action of thapsigargin, a specific inhibitor of the ER calcium pump. Mefloquine also induced a sustained influx of extra-neuronal calcium via an unknown mechanism. The transcription of key ER proteins including GADD153, PERK, GRP78, PDI, GRP94 and calreticulin were up regulated by mefloquine, suggesting that the drug induced an ER stress response. These effects appear to be related, in terms of dose effect and kinetics of action, to the acute neurotoxicity of the drug in vitro. CONCLUSIONS: Mefloquine was found to disrupt neuronal calcium homeostasis and induce an ER stress response at physiologically relevant concentrations, effects that may contribute, at least in part, to the neurotoxicity of the drug in vitro. PMID- 12848899 TI - Epithelial Na, K-ATPase expression is down-regulated in canine prostate cancer; a possible consequence of metabolic transformation in the process of prostate malignancy. AB - BACKGROUND: An important physiological function of the normal prostate gland is the synthesis and secretion of a citrate rich prostatic fluid. In prostate cancer, citrate production levels are reduced as a result of altered cellular metabolism and bioenergetics. Na, K-ATPase is essential for citrate production since the inward Na+ gradients it generates are utilized for the Na+ dependent uptake of aspartate, a major substrate for citrate synthesis. The objective of this study was to compare the expression of previously identified Na, K-ATPase isoforms in normal canine prostate, benign prostatic hyperplasia (BPH) and prostatic adenocarcinoma (PCa) using immunohistochemistry in order to determine whether reduced citrate levels in PCa are also accompanied by changes in Na, K ATPase expression. RESULTS: Expression of Na, K-ATPase alpha1 and beta1 isoforms was observed in the lateral and basolateral plasma membrane domains of prostatic epithelial cells in normal and BPH prostates. Canine kidney was used as positive control for expression of Na, K-ATPase alpha1 and gamma isoforms. The alpha1 isoform was detected in abundance in prostatic epithelial cells but there was no evidence of alpha2, alpha3 or gamma subunit expression. In advanced PCa, Na, K ATPase alpha1 isoform expression was significantly lower compared to normal and BPH glands. The abundant basolateral immunostaining observed in normal and BPH tissue was significantly attenuated in PCa. CONCLUSION: The loss of epithelial structure and function and the transformation of normal epithelial cells to malignant cells in the canine prostate have important implications for cellular metabolism and are accompanied by a down regulation of Na, K-ATPase. PMID- 12848900 TI - Role of glucocorticoids in mediating effects of fasting and diabetes on hypothalamic gene expression. AB - BACKGROUND: Fasting and diabetes are characterized by elevated glucocorticoids and reduced insulin, leptin, elevated hypothalamic AGRP and NPY mRNA, and reduced hypothalamic POMC mRNA. Although leptin replacement can reverse changes in hypothalamic gene expression associated with fasting and diabetes, leptin also normalizes corticosterone; therefore the extent to which the elevated corticosterone contributes to the regulation of hypothalamic gene expression in fasting and diabetes remains unclear. To address if elevated corticosterone is necessary for hypothalamic responses to fasting and diabetes, we assessed the effects of adrenalectomy on hypothalamic gene expression in 48-hour-fasted or diabetic mice. To assess if elevated corticosterone is sufficient for the hypothalamic responses to fasting and diabetes, we assessed the effect of corticosterone pellets implanted for 48 hours on hypothalamic gene expression. RESULTS: Fasting and streptozotocin-induced diabetes elevated plasma glucocorticoid levels and reduced serum insulin and leptin levels. Adrenalectomy prevented the rise in plasma glucocorticoids associated with fasting and diabetes, but not the associated reductions in insulin or leptin. Adrenalectomy blocked the effects of fasting and diabetes on hypothalamic AGRP, NPY, and POMC expression. Conversely, corticosterone implants induced both AGRP and POMC mRNA (with a non-significant trend toward induction of NPY mRNA), accompanied by elevated insulin and leptin (with no change in food intake or body weight). CONCLUSION: These data suggest that elevated plasma corticosterone mediate some effects of fasting and diabetes on hypothalamic gene expression. Specifically, elevated plasma corticosterone is necessary for the induction of NPY mRNA with fasting and diabetes; since corticosterone implants only produced a non significant trend in NPY mRNA, it remains uncertain if a rise in corticosterone may be sufficient to induce NPY mRNA. A rise in corticosterone is necessary to reduce hypothalamic POMC mRNA with fasting and diabetes, but not sufficient for the reduction of hypothalamic POMC mRNA. Finally, elevated plasma corticosterone is both necessary and sufficient for the induction of hypothalamic AGRP mRNA with fasting and diabetes. PMID- 12848901 TI - A nearly continuous measure of birth weight for gestational age using a United States national reference. AB - BACKGROUND: Fully understanding the determinants and sequelae of fetal growth requires a continuous measure of birth weight adjusted for gestational age. Published United States reference data, however, provide estimates only of the median and lowest and highest 5th and 10th percentiles for birth weight at each gestational age. The purpose of our analysis was to create more continuous reference measures of birth weight for gestational age for use in epidemiologic analyses. METHODS: We used data from the most recent nationwide United States Natality datasets to generate multiple reference percentiles of birth weight at each completed week of gestation from 22 through 44 weeks. Gestational age was determined from last menstrual period. We analyzed data from 6,690,717 singleton infants with recorded birth weight and sex born to United States resident mothers in 1999 and 2000. RESULTS: Birth weight rose with greater gestational age, with increasing slopes during the third trimester and a leveling off beyond 40 weeks. Boys had higher birth weights than girls, later born children higher weights than firstborns, and infants born to non-Hispanic white mothers higher birth weights than those born to non-Hispanic black mothers. These results correspond well with previously published estimates reporting limited percentiles. CONCLUSIONS: Our method provides comprehensive reference values of birth weight at 22 through 44 completed weeks of gestation, derived from broadly based nationwide data. Other approaches require assumptions of normality or of a functional relationship between gestational age and birth weight, which may not be appropriate. These data should prove useful for researchers investigating the predictors and outcomes of altered fetal growth. PMID- 12848902 TI - The Ischemic Stroke Genetics Study (ISGS) Protocol. AB - BACKGROUND: The molecular basis for the genetic risk of ischemic stroke is likely to be multigenic and influenced by environmental factors. Several small case control studies have suggested associations between ischemic stroke and polymorphisms of genes that code for coagulation cascade proteins and platelet receptors. Our aim is to investigate potential associations between hemostatic gene polymorphisms and ischemic stroke, with particular emphasis on detailed characterization of the phenotype. METHODS/DESIGN: The Ischemic Stroke Genetic Study is a prospective, multicenter genetic association study in adults with recent first-ever ischemic stroke confirmed with computed tomography or magnetic resonance imaging. Patients are evaluated at academic medical centers in the United States and compared with sex- and age-matched controls. Stroke subtypes are determined by central blinded adjudication using standardized, validated mechanistic and syndromic classification systems. The panel of genes to be tested for polymorphisms includes beta-fibrinogen and platelet glycoprotein Ia, Iba, and IIb/IIIa. Immortalized cell lines are created to allow for time- and cost efficient testing of additional candidate genes in the future. DISCUSSION: The study is designed to minimize survival bias and to allow for exploring associations between specific polymorphisms and individual subtypes of ischemic stroke. The data set will also permit the study of genetic determinants of stroke outcome. Having cell lines will permit testing of future candidate risk factor genes. PMID- 12848903 TI - Identification of a distinct class of cytoskeleton-associated mRNAs using microarray technology. AB - BACKGROUND: Interactions between mRNA and the cytoskeleton are critical for the localization of a number of transcripts in eukaryotic somatic cells. To characterize additional transcripts that may be subject to this form of regulation, we developed a two-step approach that utilizes biochemical fractionation of cells to isolate transcripts from different subcellular compartments followed by microarray analysis to examine and compare these subpopulations of transcripts in a massively-parallel manner. RESULTS: Using this approach, mRNA was extracted from the cytoskeleton-rich and the cytosolic fractions of the promyelocytic HL-60 cell line. We identify a subset of 22 transcripts that are significantly enriched in the cytoskeleton-associated population. The majority of these encode structural proteins and/or proteins known to interact with elements of the cytoskeleton. Localization required an intact actin cytoskeleton and was largely conserved upon differentiation of precursor HL-60 cells to a macrophage-like phenotype. CONCLUSIONS: We conclude that the association of transcripts with the actin cytoskeleton in somatic cells may be a critical post-transcriptional regulatory event that controls a larger class of genes than has previously been recognized. PMID- 12848904 TI - [Primary analysis on protein maps of Helicobacter pylori strains associated with gastric carcinoma]. AB - OBJECTIVE: To identify specific proteins of Helicobacter pylori (H. pylori) that associated with gastric carcinoma. METHODS: The whole-cell proteins of H. pylori were separated by two-dimensional electrophoresis (2-DE). The protein maps of four H. pylori strains associated with gastric carcinoma and nine strains that isolated from patients with non-gastric carcinoma were then compared by ImageMaster 2D v3.1. MALDI-TOF mass spectrometry was performed to identify the proteins of interest. The proteins were searched by software mascot and identified by peptide fingerprint map. RESULTS: Three proteins seemed to be associated with gastric carcinoma including acylneuraminate cytidylyltransferase with Mowse score 79 with the sequence coverage of 32%. The other two had no unambiguous protein to match. CONCLUSION: Acylneuraminate cytidylyltransferase seemed to be a specific H. pylori protein associated with the presence of gastric carcinoma. Other two were novel proteins that might be associated with gastric carcinoma. However, the mechanism needs to be explored. PMID- 12848905 TI - [Meta-analysis on the epidemiology of Helicobacter pylori infection in China]. AB - OBJECTIVE: To understand the epidemiologic features and etiology of Helicobacter pylori (H. pylori) infection in China so as to provide evidence for the development of preventive measures on H. pylori infection and related diseases. METHODS: Meta-analysis was used to evaluate the data extracted from 1990 - 2002 published papers on the epidemiology of H. pylori infection in China. RESULTS: The average infection rate of H. pylori infection in China was 58.07%, with 50% in 10 to 20 year-olds. Significant intrafamilial clustering was observed. H. pylori infection was a risk factor to develop gastrointestinal disease. CONCLUSIONS: H. pylori infections in Chinese were common and extensively distributed, which might due to close contact between family members. PMID- 12848906 TI - [In-vitro activity of rabeprazole, lansoprazole, and esomeprazole against Helicobacter pylori]. AB - OBJECTIVE: To investigate the antimicrobial activity of Pariet, Tekpron, Nexium, respectively, against Helicobacter pylori (H. pylori) in vitro. METHODS: Antimicrobial effects of these medicines were evaluated through detection of MICs for 3 H. pylori strains isolated from different countries. RESULTS: The MIC(99) contents were 2.25 mg/L, 42.5 mg/L and 360 mg/L, respectively, for the three medicines. The strains under testing exhibited the same susceptibility to each medicine. Nexium did not inhibit the bacteria under the concentration of 3.6 - 36 mg/L with more and bigger H. pylori colonies seen when compared with controls. CONCLUSIONS: The growth inhibitory activity appeared to be different among the three PPI medicines under investigation, with Rabeprazole the most potential agent of the three. Data suggested that the action of growth inhibition in vitro was resting on the characteristic of the given PPI as well as the supplements of the medicine. PMID- 12848907 TI - [Analyses on one case of severe acute respiratory syndrome 'super transmitter' and chain of transmission]. AB - OBJECTIVES: To investigate the transmission process of severe acute respiratory syndrome (SARS) and to evaluate the infectiveness of SARS patients in different periods of disease epidemics. METHODS: Standardized questionnaire was used to conduct case investigation and contact tracing by combining the field investigation and telephone interview. Transmission process, infectivity, transmission chain and contact history of SARS were studied through data analyses. RESULTS: On 25th March 2003, a 91 year old man was admitted to Hospital J in Beijing with stroke and fever. He died on 30th March. From 31st March, there was an outbreak of SARS among his contacts in the family and in the hospital he was admitted to. Contacts would include his relatives, other co-patients and health care workers in the Hospital J. Chinese Field Epidemiology Training Program trainees conducted an investigation of the outbreak. Among the 207 contacts of the index cases through different generations, there were 36 cases of SARS (attack rate 17%) patients with one death. There were 12 cases having directly contact with the index case and 13 cases with one secondary case. The transmission chains of this outbreak could clearly be depicted. All the cases had close contacts during the symptomatic period of their index patients. Among the relatives, 85% of the cases had 3 - 5-day contact with their index patients after the onset of the illnesses. There was no significant difference between the two attack rates-70% for whose who had contact with the patient before and after illness onset) and 67% for those who only had contact after the onset of the illness. Out of the 44 social acquaintances and 38 of the family members who had contacts with the index patients during the incubation period, no one was found ill. Among the close contacts at the hospital who had no protection when providing care to the patient, the attack rate was found over 80%. CONCLUSIONS: All the secondary cases of this outbreak had a history of direct and close contacts to the index patients after the onset of the illness. There was no evidence indicating that SARS cases were infectious during their incubation period. PMID- 12848908 TI - [Study on the epidemiological characteristics of severe acute respiratory syndrome in Shanxi province]. AB - OBJECTIVE: To describe the epidemiological characteristics and related factors of SARS in Shanxi in order to provide scientific basis for prevention and control of severe acute respiratory syndrome (SARS). METHODS: Data on clinically-diagnosed SARS cases reported to Shanxi Center for Disease Control and Prevention through SARS reporting system of Shanxi province and epidemiological reports were collected from early March to 20 May, 2003. The characteristics of SARS distribution in time, place and population in Shanxi were described. The epidemiological characteristics and related influential factors were analyzed with EPI info 6.0 software. RESULTS: Since the first imported SARS case was diagnosed clinically on 7 March and till 20 May in Shanxi province, the number of cumulative clinically-diagnosed SARS cases were 445 with an attack rate of 1.34/10,000. 20 deaths occurred in that period with the mortality rate 4.49%. The number of cases increased from 28 March and formed the first peak. However, the number continued to increase until 18 April to have formed the second peak. Since then, the number of cases has gradually decreased gradually. Since 19 May, there has been no clinically-diagnosed cases being reported. SARS cases were mostly seen in urban areas of the city (83.82% of the total SARS cases) with sporadic cases found in rural areas. Students and medical staff and people from 20 - 59 years of age occupied the large part of the cases. Age specific mortality rate increased with age and the male/female ratio was 1:0.87. CONCLUSION: In Shanxi province, the SARS epidemic seemed to have had the following stages: importation of the first case, gradual increase of the number of cases to reach the peak and decreasing. Case identification at early stage as well as taking measures to decrease the chance of transmission were strategically crucial for controlling the spread of SARS virus in the community. PMID- 12848909 TI - [Analysis on the trend of severe acute respiratory syndrome epidemic in Inner Mongolian Autonomous Region]. AB - OBJECTIVE: To analyse the severe acute respiratory syndrome (SARS) epidemics in Inner Mongolian Autonomous Region and to provide scientific basis for prevention and control strategies against it. METHODS: Data from legal communicable diseases surveillance reporting system was analyzed epidemiologically. RESULTS: The first SARS case was reported in Inner Mongolian Autonomous Region on March 27, 2003. Up to May 20, there were 446 cumulative SARS cases in the whole region (with 287 confirmed cases and 159 suspected cases) and 61 cumulative recovered cases had been discharged from the hospitals (56 confirmed cases and 5 suspected cases). Another 131 cases were excluded the original diagnoses of SARS including 10 confirmed cases and 121 suspected cases. 25 confirmed cases died with a mortality rate of 8.7%. Cumulatively, the number of reported cases were distributed in 30 counties in 9 prefectures. Statistical analysis on time sequence of the occurrence of cases showed that majority (67.7% of the total) of the cases concentrated in between April 13 and April 29. The number of cases had started to decrease since April 24 with an average of 5.3 cases per day between May 3 and May 8 and an average of 0.3 cases per day between May 9 and today. CONCLUSIONS: SARS epidemics in our region could be divided into three phases. The first phase fell in between March 18 and April 15 with the first case being imported, the number of cases rose sharply, covering 14 counties in 6 prefectures, having a feature of family clustering. The second phase was from April 16 to April 28, with the appearance of secondary infection, having sharp rise of the cases and spreading to 24 counties in 10 prefectures. One of the major features was that hospitals had become the important sources of secondary infection. Finally, the third phase was between April 29 and May 20, with small wave crests of cases, spreading to 38 counties in 10 prefectures with a high proportion of cases with no history of direct contact with diagnosed SARS patients. Thus, no obvious transmission chain was noticed at this phase. PMID- 12848910 TI - [Severe acute respiratory syndrome epidemiology in Sichuan province]. AB - OBJECTIVE: To explore the epidemiological characters of severe acute respiratory syndrome (SARS) in Sichuan province in order to provide evidence for prevention and control. METHODS: To generate data on SARS in Sichuan province through descriptive and analytical studies on time, geographic distribution, population, source of infection, the way of case finding, symptom, diagnosis and treatment of the cases. RESULTS: The peak of the epidemic last from April 16 to May 7. The number of cases in Luzhou and Guangyuan cities took up 60% of the total. Mobile population occupied 68% of the cases. Most of the patients were above the age of 20 with a sex ratio of 1.5:1 (m/f). 80% of the cases had a history of working in Guangdong province and recently returned to their hometowns. The main symptoms and signs of the SARS patients would include fever, cough and chest X-ray changes. CONCLUSION: All cases were imported. Fluctuation of the epidemics was mainly affected by the mobility of working population who recently returned to their hometowns. Measures concerning the prevention and control of the epidemics would mainly target on the isolation of confirmed and suspected patients who might serve as the sources of infection through setting up quarantine station, assigned hospitals and special 'fever-clinics'. PMID- 12848911 TI - [The epidemiology of neural tube defects in high-prevalence and low-prevalence areas of China]. AB - OBJECTIVE: To describe the epidemiology of neural tube defects (NTDs) in high- and low-prevalence areas of China. METHODS: Birth defects surveillance data, collected from 1992 through 1994 was analyzed. These data were collected as part of the Sino-American cooperative project on NTDs prevention. We classified NTDs as anencephaly, encephalocele, high-level and low-level spina bifida (SB) according to location of the lesion (high vs low) and whether the defect was isolated or occurred in association with other birth defects. Rates were compared in the high-prevalence (North) region and the low-prevalence (South) region, after adjusted for classification, urban and rural, season and sex, and calculated the adjusted rate of NTDs. RESULTS: Among seven hundred and eighty four NTDs cases in 326 874 recorded births (include in livebirth, stillbirth and fetal death with a gestational age of at least 20 weeks), the overall NTDs prevalence in the North was 5.57/1,000 births, and in the South was 0.88/1 000. There were also significant differences in the prevalence of anencephaly, encephalocele, high-level and low-level SB between North (0.97, 0.49, 2.75 and 1.11/1,000 birth) and South (0.36, 0.15, 0.21 and 0.14/1,000 birth) (P < 0.01), with adjusted prevalences in the North 3 - 7 times higher than those in the South. There were significant difference between urban (2.04) and rural areas (6.57/1,000 birth) in the North (P < 0.01), urban (0.52) and rural areas (0.95/1,000 birth) in the South (P < 0.05). Adjusted prevalence rates in the rural were 3 - 4 times higher than those of urban in the North and 1.6 - 1.9 times higher than in the South; The seasonal rate of high-level SB increased between September and November in the North (3.44/1,000 birth), while the seasonal rate of anencephaly decreased between September and November (0.18/1,000 birth) in the South. However there were no seasonal changes in other classified NTDs both in the South and North. CONCLUSIONS: The birth prevalence of NTDs in the North of China was the highest in the world. There were significant differences between the North and the South, urban and rural. There was seasonal change in high-level SB in the North, which was in accordance to the phenotype of NTDs. It was suggested that there might exist etiological heterogeneity among anecephalus, low- and high-level SB. PMID- 12848912 TI - [Mean level of blood pressure and rate of hypertension among people with different levels of body mass index and waist circumference]. AB - OBJECTIVE: To assess the relationship between body mass index (BMI), waist circumference (WC) and blood pressure (BP) including hypertension. METHODS: A total of 30 003 participants aged 35 - 59 from different parts of China were surveyed for cardiovascular disease risk factors in two independent cross sectional studies that were carried out in 1992 - 1994 and in 1998. Data were pooled to analyze the levels of BP and rates of hypertension with different levels of BMI and WC. RESULTS: The level of BP and rate of hypertension were significantly increasing with rising of WC among the majority of BMI groups (P value for trend < 0.05) and significantly increasing with rising of BMI in all WC groups (P value for trend < 0.05). The age adjusted hypertension rate with BMI < 24.0 kg/m(2) and WC < 85/80 cm (men/women), BMI < 24.0 kg/m(2) and WC >or= 85/80 cm (M/W), BMI 24.0 - 27.9 kg/m(2) and WC < 85/80 cm (M/W), BMI 24.0 - 27.9 kg/m(2) and WC >or= 85/80 cm (M/W), BMI >or= 28 kg/m(2) and WC >or= 85/80 cm (M/W) were 16.5%, 29.8%, 29.6%, 39.2% and 57.5% for men and 14.1%, 20.6%, 24.7%, 30.3% and 43.3% for women, respectively. CONCLUSION: Independent influence of BMI or WC on BP was noticed. Keeping BMI and WC at normal level might effectively prevent the occurrence of hypertension. PMID- 12848913 TI - [Study on mortality, incidence and risk factors of stroke in a cohort of elderly in Xi'an, China]. AB - OBJECTIVE: To prospectively study the relationship between risk factors and stroke incidence and mortality in the Chinese elderly. METHODS: An analytic study in a cohort population of 1,268 male retired cadres in a military setting. A health-screening program was carried out for all cadres aged 55 years or older in Xi'an in February 1987. Baseline data on smoking, cholesterol, triglyceride, blood pressure, body mass index (BMI), histories of hypertension, hyperlipidemia and coronary heart disease (CHD), stroke, diabetes and family histories of cardiovascular disease were investigated. Main outcome measures were stroke incidence, stroke and all-cause mortality. RESULTS: The cohort was followed up until June 30, 2001 and a total number of follow-up person-year was 15,546. During the follow-up period, there were 113 new stroke cases identified and the adjusted incidence was 727 per 100,000 person-year. Forty-five deaths were due to stroke and the adjusted mortality was 289 per 100,000 person-year. Using Cox model analysis, after adjustment on age, total cholesterol, triglyceride, smoking, drinking and physical exercise, we noticed that the systolic pressure, BMI, history of CHD and hyperlipidemia were independent risk factors for stroke incidence and morality. CONCLUSION: The incidence and mortality of stroke in this cohort were lower than those in the same age group of general population. Monitoring and controlling body mass index and blood pressure level seemed to be important factors for the prevention of stroke in the elderly. PMID- 12848914 TI - [A case-control study on the risk factors for road injury]. AB - OBJECTIVE: To investigate the risk factors of road injury. METHODS: Case-control study was used. From November 2001 to August 2002, 406 drivers who had 438 drivers who had not experienced a motor vehicle crash in Huanggu district, Shenyang city were recruited by randomly selection on time of day, day of week and site in the same period at same district. Face to face interviews with drivers were conducted according to a highly structured questionnaire covering the circumstances of the current trip, usual behavior and background characteristics of the drivers and the condition of motor vehicles. Stanford sleepiness scale and Epworth sleepiness scale were used to quantify acute and chronic sleepiness respectively. RESULTS: Increased risk was associated with drivers who identified themselves as having chronic doziness (OR = 1.98, 95% CI: 1.26 - 3.12). Increase in risk was associated with measures of acute tiredness, but without statistical significance (OR = 2.38, 95% CI: 0.89 - 6.31). Comparing to permanent daytime work pattern, rotating shifts or permanent night-work pattern increased the risk of crash (OR = 2.09, 95% CI: 1.48 - 2.94). The risk of motor vehicle crash among the drivers who drank alcohol in the previous 6 hours was 3.59 times (95% CI: 1.13 - 11.39) of those drivers who did not drink. Driving violations also contributed to the increased risk of crash (OR = 1.73, 95% CI: 1.22 - 2.46). CONCLUSION: Factors as chronic doziness, rotating shifts or permanent night-work pattern, driving under alcohol impairment, violation of motor vehicle regulation all significantly increased the risk of road injury. Acute sleepiness might serve as a potential risk factor for road injury. PMID- 12848915 TI - [The application of indirect immuno-fluorescence assay in the diagnosis of severe acute respiratory syndrome]. AB - OBJECTIVE: To explore the temporal profile of serum antibody against coronavirus in patients with severe acute respiratory syndrome (SARS), and to evaluate the reliability of indirect immuno-fluorescence assay (IFA) in the diagnosis of SARS. METHODS: Clinically confirmed SARS patients, suspected SARS patients, and controls were included in the study. IFA was used to detect the serum antibody against SARS coronavirus. General information about the subjects was collected using a standard questionnaire. RESULTS: The positive rates of specific IgG and IgM against SARS virus within 10 days after onset of the disease were 55.1% and 16.3% respectively and then increased up to 89.8% for IgG and 65.3% for IgM. After 25 days of the onset of the disease, 90.9% patients became positive for both IgG and IgM. Results from chi-square for trend test revealed that the positive rates of both IgG and IgM increased with time (chi(2) for trend = 16.376, P = 0.00005 for IgG; chi(2) for trend = 28.736, P = 0.00000 for IgM). Sensitivity, specificity and agreement value of IFA regarding the diagnosis of SARS were all higher than 90%. CONCLUSION: IFA can be used to assist diagnosis of SARS after 10 days of the onset of disease. PMID- 12848916 TI - [Study on vector ability of Nosopsyllus wualis leizhouensis in the transmission of plague]. AB - OBJECTIVE: To find out the vector ability and function of Nosopsyllus wualis leizhouensis in the transmitting plague. METHODS: In T: 19 degrees C +/- 1 degrees C, RH: 85% +/- 5%, data regarding the vector ability as cluster spreading, single flea spreading, single flea transmitting plague to single animal, formative bacterial embolus and infection fleas life-span through experiments was gathered. RESULTS: The rate of infection on fleas was 94.64%, with 100% transmission rate of colony to spread, and 30% from single flea spreading to single animal. In the experiment of single flea transmission, all of the 388 rattus loseas were bitten by the fleas with bacterial, but only 9 animals were characteristically infected with the transmission potential, vector efficiency, survival potential of embolus, vector index as 0.360, 0.257, 0.868 and 0.223 respectively. The mean survive days of infected flea feed with blood were 17.58 (1 - 58), and the mean survive days of hunger infected flea were 7.25 (1 - 16). Formative bacterial embolus days were 8.80 (2 - 16) and the rate of embolus flea was 78.12%. CONCLUSION: Nosopsyllus wualis leizhouensis could serve as vector and important in the mode of plague transmittion. PMID- 12848917 TI - [Sequences analysis of cytochrome C oxidase subunit I gene in Aedes albopictus from different geographic strains in China]. AB - OBJECTIVE: To compare the sequences of cytochrome C oxidase subunit I gene (COI) in Aedes albopictus from different geographic strains in China and to discuss the differences in susceptibility among different geographic strains to dengue virus (DV). METHODS: COI was amplified with polymerase chain reaction method and sequenced from its genomic DNA. Molecular phylogenetic trees were constructed with Neighbor-Joining method. RESULTS: Sequence length of COI fragment in each geographic strains was 415 bp. The rates of shift and reverse of base pairs in Simao strain were 1.93% and 0.24% respectively. The rate of shift in Mawei and Nanning strains was 0.48%. The analyses of phylogenetic of COI sequences showed that there was close relationship between Simao strain in Yunnan and Mawei strain in Guizhou and between Mawei strain and Nanning strain in Guangxi. CONCLUSIONS: The susceptibility was widely related to many factors including genetic and environmental ones. COI in Aedes albopictus from different geographic strains in China belonged to the same gene type. There were no direct correlations between COI gene type in different geographic strains and susceptibility to DV. PMID- 12848918 TI - [Dynamic expression of antigen MG7 in the gastric cancer and gastric precancerous lesions]. AB - OBJECTIVE: To study the kinetics of MG7 expression in the process of gastric cancer development. METHODS: The expression level of antigen MG7 on gastric mucosa in 406 cases was determined by immunohistochemical techniques. The classification of intestinal metaplasia of gastric mucosa was determined by histochemistry techniques on gastric mucosa in 82 cases. RESULTS: The positive rates of MG7 expression in normal gastric mucosa, intestinal metaplasia and dysplasia of gastric mucosa and gastric cancer all increased gradually (P < 0.01). The positive rates of MG7 expression in superficial gastritis, atrophic gastritis and gastric cancer increased in sequence (P < 0.01). The positive rate of antigen MG7 expression in III intestinal metaplasia of gastric mucosa was significantly different with I and II intestinal metaplasia (P < 0.05). CONCLUSIONS: MG7 was quite specific in gastric cancer thus could be used as a good index in the screening of gastric cancer. Patients with III intestinal metaplasia of gastric mucosa, atrophic gastritis and dysplasia of gastric mucosa should be closely followed in order to improve the early detection on gastric cancer. It seemed that MG7 was clinically valuable in the dynamic follow-up of gastric precursors. PMID- 12848919 TI - [Study on candidate genes of benazepril related cough in Chinese hypertensives]. AB - OBJECTIVE: To investigate the associations between angiotensin converting enzyme inhibitors (ACEIs) related cough and ACE I/D and bradykinin beta(2) receptor (BDKRB2) C/T polymorphism. METHODS: A case-control study, nested in a 3-year community-based postmarketing surveillance of benazepril in 1 831 Chinese hypertensives was carried out. Three hundred and fifty-one cases having suffered benazepril related cough were identified and genotyped. Genotyped controls were selected through a stratified sampling design by age, sex and kidney function status. RESULTS: The allele frequencies in cases were I 65.4%, D 34.6% and T 53.0%, C 47.0% and the genotype frequencies were II 42.2%, ID 46.4%, DD 11.4% (ACE) and CC 21.6%, CT 50.9%, TT 27.6% (BDKRB2), respectively. Genotype frequencies were both in Hardy-Weinberg equilibrium. According to stratified analyses by sex, kidney function status and age, no association was found between BDKRB2 C/T polymorphism and cough. For ACE I/D polymorphism, in men with decompensated kidney function, patients with ID or DD genotype having 4.805 times the risk of those with II genotype in developing cough. In women aged 35 to 49 years with normal or compensated kidney function, the OR of DD genotype was 5.128. No associations were detected in other subgroups. CONCLUSION: It was suggested that kidney function status and some specific characteristics surrogated by age and sex had modified the effect of ACE I/D variant on cough. PMID- 12848920 TI - [Study on the correlation of between infection, inflammation and coronary artery disease]. AB - OBJECTIVE: Recently studies showed infections of Chlamydia pneumoniae (Cp), Helicobacter pylori (Hp) and cytomegalovirus (CMV) played roles in the development of atherosclerosis. The aim of this study was to study relationship between infection of Cp, Hp and CMV, systemic inflammation and coronary artery disease (CAD). METHODS: Fourty-five patients with at least one coronary artery stenosis > 50% and 33 control subjects with negative coronary angiography were recruited for this case-control study from May 2000 to October 2001. Antibodies against Cp, Hp and CMV were measured and serum C-reactive protein (CRP) levels determined for each case. CRP level > 0.8 mg/dl was defined at elevated CRP level. RESULTS: The prevalence of Cp IgG, Hp IgG or Hp IgA antibody was associated with CAD (P = 0.017, P < 0.001, P = 0.009). After adjustment for age, gender, smoking, hypertension, hyperlipidemia and diabetes, the association was still seen. Mean CRP value was significantly higher in patients with CAD, compared to those without CAD (P < 0.001). Multivariate analysis showed statistical significance (P = 0.03). Elevated levels of CRP were found to be an important parameter for CAD (P = 0.032). The prevalence of Cp IgG antibody, Hp IgG and CMV IgG antibody all showed no association with elevated levels of CRP (P = 0.391, P = 0.253, P = 0.724). The ratio of elevated levels of serum basic CRP in the group with IgG antibodies to 3 pathogens was 32.1% while in the group with IgG antibodies to or= 60 years group vs < 60 years group P = 0.0006, BUN >or= 28.6 mmol/L vs < 28.6 mmol/L group P = 0.0415, SCr >or= 884 micro mol/L vs < 884 micro mol/L group P = 0.0146, value of plasma calcium multiplied by plasma phosphate >or= 40 vs < 40 group P = 0.0464, KT/V >or= 1.30 vs < 1.30 group P = 0.0215). CONCLUSIONS: The mortality related prognostic factors of maintained hemodialysis patients seemed to include age, BUN, SCr, value of plasma calcium multiplied by plasma phosphate, and KT/V, which were all tested during the third dialysis month. Among them, age and the value of plasma calcium multiplied by plasma phosphate were risk factors of death, while the others were protective factors. PMID- 12848922 TI - [A case-control study on the risk factor of perinatals' congenital malformations in seven cities of Guangxi]. AB - OBJECTIVE: To investigate the possible risk factors of congenital malformations in cities of Guangxi. METHODS: A case-control study was carried out on 281 cases of congenital malformations and 730 controls. Analysis of simple factor and multiple factors unconditional logistic regression were done. RESULTS: The analysis of simple factor and multiple factors showed that main risk factors of congenital malformations as multiple pregnancies (OR = 2.6), pregnancy complications (OR = 3.2), exposure to chemical substances before or during pregnancy (OR = 3.0), taking sedatives (OR = 10.2), hormone drug (OR = 9.4) or Chinese herbal medicines (OR = 2.5) during the early stage of pregnancy, mothers' blood type as AB (OR = 3.5) or A (OR = 2.2), mothers' emotion being nervous and melancholy (OR = 2.6), mothers' occupation being workers (OR = 3.8) or peasants (OR = 3.0), fathers' exposure to noise (OR = 5.7) or suffering from chronic diseases (OR = 2.8). CONCLUSIONS: Some risk factors were identified as having important effect on perinatal congenital malformations, including taking sedatives, hormone drug or Chinese herbal medicines during the early stage of pregnancy, mothers' emotion being nervous and melancholy, multiple pregnancies, pregnancy complications, exposure to chemical substances before or during pregnancies, mothers' blood type as AB or A, mothers' occupation being workers or peasants, fathers' exposure to noise or suffering from chronic diseases. PMID- 12848923 TI - [An effective method to reduce bias between two compared groups: propensity score]. AB - OBJECTIVE: Through introduction of principal theory and algorithm of propensity score to design SAS macro programs for binary data. METHODS: Propensity score method was used to compare the differences of character variables between two groups, and the association of DNR (Do Not Resuscitate) with the mortality of congestive heart failure was evaluated with different methods. RESULTS: Significant differences among the character variables between two groups were effectively balanced with stratification or matching method. The odds ratios of DNR with the in-hospital mortality rate of congestive heart failure were estimated identical with different algorithms and to find that the association of DNR to in-hospital mortality was highly significant. CONCLUSION: Propensity score was a good algorithm that could be used to analyze any kind of observational data for matching the effects among the character variables. PMID- 12848924 TI - Retrograde transport redux. AB - Trafficking of trophic factors in axons can occur in a retrograde and anterograde direction. Recent findings bring further support for a vesicle-based process for retrograde transport but raise new questions that need to be pursued. Unraveling the exact mechanisms that account for retrograde transport of neurotrophins and their receptors will reveal the cellular requirements for propagating trophic signals over long distances. PMID- 12848925 TI - Visual system plasticity begins in the retina. AB - Visual experience is known to induce developmental plasticity in visual cortex; now, Tian and Copenhagen report that experience regulates the development of retinal circuitry itself. Both pruning of retinal ganglion dendrites into ON or OFF sublamina and the emergence of pure ON versus OFF responses require visual experience. PMID- 12848926 TI - "The little engine that could": voltage-dependent Na(+) channels and the subthalamic nucleus. AB - The most effective treatment for late-stage Parkinson's disease is to electrically stimulate the subthalamic nucleus (STN) at high frequencies. Why this strategy works is unclear. The work by Do and Bean shows that the Na channels in STN neurons have distinctive features--like resurgence--that regulate their spiking behavior, providing new insights into the mechanism of DBS. PMID- 12848927 TI - NPY and the regulation of behavioral development. AB - Neuropeptide Y is implicated in the regulation of feeding in vertebrates, but recent studies in transgenic mice are contradictory. In this issue of Neuron, Wu et al. show a dual role for the Drosophila NPY (dNPF) in the developmental regulation of larval foraging and social behaviors, demonstrating a conserved role for this peptide in complex behaviors. PMID- 12848928 TI - Gating of GIRK channels: details of an intricate, membrane-delimited signaling complex. AB - G protein-coupled inwardly rectifying potassium channels (GIRK/Kir3) are important elements in controlling cellular excitability. In recent years, tremendous progress has been made toward understanding various components involved in channel activation, modulation, and signaling specificity. In this review, we summarize these recent findings and attempt to put them in context with recently available structural data. PMID- 12848929 TI - Basic helix-loop-helix factors in cortical development. AB - Transcription factors with bHLH motifs modulate critical events in the development of the mammalian neocortex. Multipotent cortical progenitors are maintained in a proliferative state by bHLH factors from the Id and Hes families. The transition from proliferation to neurogenesis involves a coordinate increase in the activity of proneural bHLH factors (Mash1, Neurogenin1, and Neurogenin2) and a decrease in the activity of Hes and Id factors. As development proceeds, inhibition of proneural bHLH factors in cortical progenitors promotes the formation of astrocytes. Finally, the formation of oligodendrocytes is triggered by an increase in the activity of bHLH factors Olig1 and Olig2 that may be coupled with a decrease in Id activity. Thus, bHLH factors have key roles in corticogenesis, affecting the timing of differentiation and the specification of cell fate. PMID- 12848930 TI - All-optical histology using ultrashort laser pulses. AB - As a means to automate the three-dimensional histological analysis of brain tissue, we demonstrate the use of femtosecond laser pulses to iteratively cut and image fixed as well as fresh tissue. Cuts are accomplished with 1 to 10 microJ pulses to ablate tissue with micron precision. We show that the permeability, immunoreactivity, and optical clarity of the tissue is retained after pulsed laser cutting. Further, samples from transgenic mice that express fluorescent proteins retained their fluorescence to within microns of the cut surface. Imaging of exogenous or endogenous fluorescent labels down to 100 microm or more below the cut surface is accomplished with 0.1 to 1 nJ pulses and conventional two-photon laser scanning microscopy. In one example, labeled projection neurons within the full extent of a neocortical column were visualized with micron resolution. In a second example, the microvasculature within a block of neocortex was measured and reconstructed with micron resolution. PMID- 12848931 TI - The regulation of glucose metabolism by HIF-1 mediates a neuroprotective response to amyloid beta peptide. AB - It is frequently argued that both amyloid beta (Abeta) and oxidative stress are involved in the pathogenesis of Alzheimer's disease (AD). We show here that clonal nerve cell lines and primary cortical neurons that are resistant to Abeta toxicity have an enhanced flux of glucose through both the glycolytic pathway and the hexose monophosphate shunt. AD brain also has increased enzymatic activities in both pathways relative to age-matched controls. The Abeta-induced changes in glucose metabolism are due to the activation of the transcription factor hypoxia inducible factor 1 (HIF-1). As a result of Abeta-induced changes in glucose metabolism, Abeta-resistant cells are more readily killed by glucose starvation and by classes of antipsychotic drugs that inhibit glucose uptake. PMID- 12848932 TI - Evidence in support of signaling endosome-based retrograde survival of sympathetic neurons. AB - The mechanism by which target-derived Nerve Growth Factor (NGF) signaling is propagated retrogradely, over extremely long distances, to cell bodies to support survival of neurons is unclear. Here we show that survival of sympathetic neurons supported by NGF on distal axons requires the kinase activity of the NGF receptor, TrkA, in both distal axons and cell bodies. In contrast, disruption of TrkA activity exclusively in proximal axonal segments affects neither retrograde NGF-TrkA signaling in cell bodies nor neuronal survival. Ligand-receptor internalization is necessary for survival of neurons supported by NGF on distal axons. Furthermore, antibody neutralization experiments indicate that retrogradely transported NGF, within cell bodies, is critical for neuronal survival but not for growth of distal axons. Taken together, our results indicate that retrogradely transported NGF-TrkA complexes promote sympathetic neuron survival. PMID- 12848933 TI - NGF signaling in sensory neurons: evidence that early endosomes carry NGF retrograde signals. AB - Target-derived NGF promotes the phenotypic maintenance of mature dorsal root ganglion (DRG) nociceptive neurons. Here, we provide in vivo and in vitro evidence for the presence within DRG neurons of endosomes containing NGF, activated TrkA, and signaling proteins of the Rap1/Erk1/2, p38MAPK, and PI3K/Akt pathways. Signaling endosomes were shown to be retrogradely transported in the isolated sciatic nerve in vitro. NGF injection in the peripheral target of DRG neurons increased the retrograde transport of p-Erk1/2, p-p38, and pAkt in these membranes. Conversely, NGF antibody injections decreased the retrograde transport of p-Erk1/2 and p-p38. Our results are evidence that signaling endosomes, with the characteristics of early endosomes, convey NGF signals from the target of nociceptive neurons to their cell bodies. PMID- 12848934 TI - Visual stimulation is required for refinement of ON and OFF pathways in postnatal retina. AB - ON and OFF pathways separately relay increment and decrement luminance signals from retinal bipolar cells to cortex. ON-OFF retinal ganglion cells (RGCs) are activated via synaptic inputs onto bistratified dendrites localized in the ON and OFF regions of the inner plexiform layer. Postnatal maturational processes convert bistratifying ON-OFF RGCs to monostratifying ON and OFF RGCs. Although visual deprivation influences refinement of higher visual centers, no previous studies suggest that light regulates either the development of the visual-evoked signaling in retinal ON and OFF pathways, nor pruning of bistratified RGC dendrites. We find that dark rearing blocks both the maturational loss of ON-OFF responsive RGCs and the pruning of dendrites. Thus, in retina, there is a previously unrecognized, pathway-specific maturation that is profoundly affected by visual deprivation. PMID- 12848935 TI - FRET two-hybrid mapping reveals function and location of L-type Ca2+ channel CaM preassociation. AB - L-type Ca(2+) channels possess a Ca(2+)-dependent inactivation (CDI) mechanism, affording feedback in diverse neurobiological settings and serving as prototype for unconventional calmodulin (CaM) regulation emerging in many Ca(2+) channels. Crucial to such regulation is the preassociation of Ca(2+)-free CaM (apoCaM) to channels, facilitating rapid triggering of CDI as Ca(2+)/CaM shifts to a channel IQ site (IQ). Progress has been hindered by controversy over the preassociation site, as identified by in vitro assays. Most critical has been the failure to resolve a functional signature of preassociation. Here, we deploy novel FRET assays in live cells to identify a 73 aa channel segment, containing IQ, as the critical preassociation pocket. IQ mutations disrupting preassociation revealed accelerated voltage-dependent inactivation (VDI) as the functional hallmark of channels lacking preassociated CaM. Hence, the alpha(1C) IQ segment is multifunctional-serving as ligand for preassociation and as Ca(2+)/CaM effector site for CDI. PMID- 12848936 TI - Subthreshold sodium currents and pacemaking of subthalamic neurons: modulation by slow inactivation. AB - Neurons of the subthalamic nucleus (STN) are spontaneously active. By voltage clamping dissociated rat STN neurons with their own firing patterns, we found that pacemaking is driven by two kinds of subthreshold sodium current: a steady state "persistent" sodium current and a dynamic "resurgent" sodium current, which promotes rapid firing by flowing immediately after a spike. These currents are strongly regulated by a process of slow inactivation that is active at physiological firing frequencies. Slow inactivation of the pacemaking sodium currents promotes a constant frequency of tonic firing in the face of small, steady changes in input and constitutes a form of adaptation at the single-cell level. Driving cells at a high rate (75 Hz) produced pronounced slow inactivation (60%-70%) of resurgent, persistent, and transient components of sodium current. This inactivation is likely to contribute to effects of clinical deep-brain stimulation on STN excitability. PMID- 12848937 TI - Light adaptation through phosphoinositide-regulated translocation of Drosophila visual arrestin. AB - Photoreceptor cells adapt to bright or continuous light, although the molecular mechanisms underlying this phenomenon are incompletely understood. Here, we report a mechanism of light adaptation in Drosophila, which is regulated by phosphoinositides (PIs). We found that light-dependent translocation of arrestin was defective in mutants that disrupt PI metabolism or trafficking. Arrestin bound to PIP(3) in vitro, and mutation of this site delayed arrestin shuttling and resulted in defects in the termination of the light response, which is normally accelerated by prior exposure to light. Disruption of the arrestin/PI interaction also suppressed retinal degeneration caused by excessive endocytosis of rhodopsin/arrestin complexes. These findings indicate that light-dependent trafficking of arrestin is regulated by direct interaction with PIs and is required for light adaptation. Since phospholipase C activity is required for activation of Drosophila phototransduction, these data point to a dual role of PIs in phototransduction. PMID- 12848938 TI - Contribution of Drosophila DEG/ENaC genes to salt taste. AB - The ability to detect salt is critical for the survival of terrestrial animals. Based on amiloride-dependent inhibition, the receptors that detect salt have been postulated to be DEG/ENaC channels. We found the Drosophila DEG/ENaC genes Pickpocket11 (ppk11) and Pickpocket19 (ppk19) expressed in the larval taste sensing terminal organ and in adults on the taste bristles of the labelum, the legs, and the wing margins. When we disrupted PPK11 or PPK19 function, larvae lost their ability to discriminate low concentrations of Na(+) or K(+) from water, and the electrophysiologic responses to low salt concentrations were attenuated. In both larvae and adults, disrupting PPK11 or PPK19 affected the behavioral response to high salt concentrations. In contrast, the response of larvae to sucrose, pH 3, and several odors remained intact. These results indicate that the DEG/ENaC channels PPK11 and PPK19 play a key role in detecting Na(+) and K(+) salts. PMID- 12848939 TI - Developmental control of foraging and social behavior by the Drosophila neuropeptide Y-like system. AB - Animals display stereotyped behavioral modifications during development, but little is known about how genes and neural circuits are regulated to turn on/off behaviors. Here we report that Drosophila neuropeptide F (dNPF), a human NPY homolog, coordinates larval behavioral changes during development. The brain expression of npf is high in larvae attracted to food, whereas its downregulation coincides with the onset of behaviors of older larvae, including food aversion, hypermobility, and cooperative burrowing. Loss of dNPF signaling in young transgenic larvae led to the premature display of behavioral phenotypes associated with older larvae. Conversely, dNPF overexpression in older larvae prolonged feeding, and suppressed hypermobility and cooperative burrowing behaviors. The dNPF system provides a new paradigm for studying the central control of cooperative behavior. PMID- 12848940 TI - Synaptic transmission and plasticity in the absence of AMPA glutamate receptor GluR2 and GluR3. AB - The AMPA glutamate receptor (AMPAR) subunits GluR2 and GluR3 are thought to be important for synaptic targeting/stabilization of AMPARs and the expression of hippocampal long-term depression (LTD). In order to address this hypothesis genetically, we generated and analyzed knockout mice deficient in the expression of both GluR2 and GluR3. We show here that the double knockout mice are severely impaired in basal synaptic transmission, demonstrating that GluR2/3 are essential to maintain adequate synaptic transmission in vivo. However, these mutant mice are competent in establishing several forms of long-lasting synaptic changes in the CA1 region of the hippocampus, including LTD, long-term potentiation (LTP), depotentiation, and dedepression, indicating the presence of GluR2/3-independent mechanisms of LTD expression and suggesting that AMPA receptor GluR1 alone is capable of various forms of synaptic plasticity. PMID- 12848941 TI - Inhibitory and excitatory mechanisms underlying auditory responses to learned vocalizations in the songbird nucleus HVC. AB - Speech and birdsong require auditory feedback for their development and maintenance, necessitating precise auditory encoding of vocal sounds. In songbirds, the telencephalic song premotor nucleus HVC contains neurons that respond highly selectively to the bird's own song (BOS), a property distinguishing HVC from its auditory afferents. We examined the contribution of inhibitory and excitatory synaptic inputs to BOS-evoked firing in those HVC neurons innervating a pathway essential for audition-dependent vocal plasticity. Using in vivo intracellular techniques, we found that G protein-coupled, potassium-mediated inhibition, tuned to the BOS, interacts with BOS-tuned excitation through several mechanisms to shape neuronal firing patterns. Furthermore, in the absence of this inhibition, the response bias to the BOS increases, reminiscent of cancellation mechanisms in other sensorimotor systems. PMID- 12848942 TI - An HLA-peptide mimics organ-specific antigen in autoimmune uveitis: its role in pathogenesis and therapeutic induction of oral tolerance. AB - Autoimmune uveitis is a sight threatening disease, which is conventionally treated with immunosuppressive medication. New treatment strategies include immunological approaches and aim at antigen specificity like oral tolerance. A peptide from the sequence of certain HLA-class I molecules plays a central role in the pathogenesis. When T cells recognize the HLA-peptide and are activated they are enabled to pass the blood-retina barrier. In the eye they recognize a cross-reactive organ-specific peptide and cause inflammation, which presents as uveitis. Here, we used the HLA-peptide as oral tolerogen to treat uveitis patients in an open study. All patients showed a positive therapeutic response and could reduce their long-lasting conventional immunosuppressive treatment. We did not observe any side effects. Moreover, side effects from conventional therapy could be reduced significantly. PMID- 12848943 TI - Functional genomics in the study of autoimmune diseases. AB - Functional genomics can be defined as a series of new biotechnological tools based on whole-genome information that can be used to understand basic mechanisms of gene regulation, structure and function. Most of these tools have been used and modeled in research of lower organisms such as the yeast C. Elegans. The time has come for their use in human genome research and there is hope that the new technologies and possibilities will provide with new knowledge on disease pathogenesis and the development of new drugs. PMID- 12848944 TI - Systemic sclerosis: current views of its pathogenesis. AB - Systemic sclerosis (SSc) is an autoimmune disorder of unknown etiology characterized by severe and often progressive cutaneous and visceral fibrosis, pronounced alterations in the microvasculature, and numerous cellular and humoral immune abnormalities. Clinically, SSc is very heterogeneous, encompassing a spectrum ranging from mild limited forms of skin sclerosis with minimal internal organ involvement to severe skin and multiple internal organ fibrosis. Mortality and morbidity in SSc are very high and are directly related to the extent of the fibrotic and microvascular alterations. A better understanding of the pathogenesis of this incurable disorder will help to better target and design effective therapy in the future. PMID- 12848945 TI - Autoreactive CD4(+) T cells to beta(2)-glycoprotein I in patients with antiphospholipid syndrome. AB - Antiphospholipid syndrome (APS) is characterized by recurrent thrombosis and intrauterine fetal loss in association with antiphospholipid antibodies (aPL). We have recently identified autoreactive CD4(+) T cells to beta(2)-glycoprotein I (beta(2)GPI) that promote aPL production in APS patients. beta(2)GPI-specific CD4(+) T cells preferentially recognize the antigenic peptide containing the major phospholipid-binding site in the context of DRB4*0103 (DR53). T-cell receptor beta chains of beta(2)GPI-specific T cells are highly restricted and mainly utilize rearranged Vbeta7 or Vbeta8 gene segments. T-cell helper activity that stimulates B cells to produce anti-beta(2)GPI antibodies is mediated through IL-6 and CD40-CD40 ligand engagement. beta(2)GPI-specific T cells respond to reduced beta(2)GPI and recombinant beta(2)GPI fragments produced in bacteria, but not to native beta(2)GPI, indicating that the epitopes recognized by beta(2)GPI specific T cells are apparently cryptic. Activation of beta(2)GPI-specific T cells resulting in production of pathogenic anti-beta(2)GPI antibodies can be induced by the exposure to cryptic peptides of beta(2)GPI. Finally, beta(2)GPI specific T cell is a reasonable target of potential therapeutic strategies that selectively suppress pathogenic aPL production in APS patients. PMID- 12848946 TI - Giant cell arteritis. AB - Giant cell arteritis is the most common vasculitis in Caucasians. The aetiology of this disease remains uncertain. This article reviews some of the recent work in epidemiology and pathology in this field, with particular regard to the immunohistochemical findings in temporal artery biopsy specimens. The possible correlation between clinical features and biopsy specimen histology is discussed, and a model of pathogenesis is presented. PMID- 12848947 TI - Proinsulin-a pathogenic autoantigen in type 1 diabetes. AB - Type 1 diabetes (T1D) is an autoimmune disease characterised by immunity to pancreatic beta-cell autoantigens, associated with beta-cell destruction leading to insulin deficiency and hyperglycaemia. The rigorous definition of an autoimmune disease requires evidence that an autoantigen elicits pathological immune responses. Using criteria for the pathogenicity of an autoantigen, we examine the evidence for proinsulin as an autoantigen in T1D. We conclude that proinsulin satisfies these criteria. As a corollary, proinsulin is a potential immunotherapeutic tool for the prevention of T1D. PMID- 12848948 TI - Antinuclear antibodies specific for primary biliary cirrhosis. AB - The serological hallmark of primary biliary cirrhosis (PBC) is the presence of antimitochondrial antibodies. However, antinuclear antibodies (ANA) are also detectable in approximately 50% of subjects with PBC. Most clinical laboratories use indirect immunofluorescence microscopy to detect ANA and two labeling patterns that predominate in PBC are 'punctate nuclear rim' and 'multiple nuclear dots.' Work over the past several years has shown that antibodies giving these patterns most often recognize nuclear pore membrane protein gp210 and nuclear body protein sp100, respectively. These ANA are highly specific for PBC and detected in approximately 25% of patients. Less frequently, ANA apparently unique to PBC recognize other proteins of the nuclear envelope and nuclear bodies. While antibodies against gp210, sp100 and some other nuclear proteins are very specific to PBC and may therefore be useful diagnostic markers, their connection to pathogenesis remains to be elucidated. PMID- 12848949 TI - Parvovirus B19 infection and autoimmune disease. AB - Human parvovirus B19 infections may cause a widespread benign and self-limiting disease in children and adults, known as erythema infectiosum or fifth disease. A variety of further manifestations are associated with the infection such as arthralgias, arthritis, leukopenia and thrombocytopenia, anemia and vasculitis, spontaneous abortion and hydrops fetalis in pregnant women. Both in children and adults parvovirus B19 infections have been frequently implicated as a cause or trigger of various forms of autoimmune diseases affecting joints, connective tissue and large and small vessels. In addition, autoimmune neutropenia, thrombocytopenia and hemolytic anemia are known as sequelae of B19 infection. The molecular basis of the autoimmune phenomena and resultant pathogenesis is unclear. The involvement of molecular mimicry between cellular and viral proteins, the induction of enhanced cytokine production via the viral transactivator protein NS1 and the phospholipase A2-like activity of the capsid protein VP1 may contribute to the induction of autoimmune reactions. All the known data and the potential mechanisms involved in the pathogenesis will be discussed in this review. PMID- 12848950 TI - Management of glucocorticoid induced osteoporosis in premenopausal women with autoimmune disease. AB - Numerous inflammatory rheumatic diseases occurring in premenopausal women require the use of high doses of glucocorticoids (GC). It was believed for many years that premenopausal women were, at least to some extent, protected from bone loss associated with GC therapy. However, epidemiological studies performed in premenopausal women with systemic lupus erythematosus, demonstrate that these patients have lower bone mineral density as compared to age-matched controls. This is explained in part by the underlying disease and in part by treatment with GC. The American College of Rheumatology recommends life style adaptation, supplementation with calcium and vitamin D in patients receiving, or initiating therapy with >/=5 mg equivalent prednisone/day. Bisphosphonates are recommended, but they should be used with caution in young women as they cross the placenta and can affect skeletal remodeling in the foetus. Bisphosphonates have a prolonged terminal half-life and data on their safety extends to 10 years. It is therefore critical to inform premenopausal women about the risks of bisphosphonates and to recommend bisphosphonates with shorter terminal half-life. PMID- 12848951 TI - The control of T cell activation vs. tolerance. AB - B7 costimulators and the T cell growth factor IL-2 are important stimuli for the activation of T lymphocytes and the development of effective immune responses. Recent studies show that the same signals promote the development of regulatory T cells or the apoptotic death of activated T cells, and thus, function to terminate immune responses and maintain self-tolerance. The balance between the different outcomes of the same signals may be determined by the magnitude of these signals and the presence or absence of concomitant stimuli, such as those generated during innate immune responses to microbes. PMID- 12848952 TI - The epidemiology of autoimmune diseases. AB - Autoimmune diseases are among the leading causes of death among young and middle aged women in the United States. Incidence rates vary among the autoimmune diseases, with estimates ranging from less than one newly-diagnosed case of systemic sclerosis to more than 20 cases of adult-onset rheumatoid arthritis per 100,000 person-years. Prevalence rates range from less than 5 per 100,000 (e.g. chronic active hepatitis, uveitis) to more than 500 per 100,000 (Grave disease, rheumatoid arthritis, thyroiditis). At least 85% of thyroiditis, systemic sclerosis, systemic lupus erythematosus, and Sjogren disease patients are female. Although most diseases can occur at any age, some diseases primarily occur in childhood and adolescence (e.g. type 1 diabetes), in the mid-adult years (e.g. myasthenia gravis, multiple sclerosis), or among older adults (e.g. rheumatoid arthritis, primary systemic vasculitis). Ethnic and geographic differences in incidence of specific autoimmune diseases have been documented, but specific groups may be at higher risk for some diseases and lower risk for other diseases. The incidence of type 1 diabetes increased but the rates of rheumatoid arthritis declined over the past 40 years. Thus although there are commonalities, there are also important demographic differences between diseases. Disease-specific research, as well as studies that focus on potentially related diseases, needs to be conducted. PMID- 12848953 TI - Cryptic infection and autoimmunity. AB - Infection as an environmental factor in autoimmunity has long been recognized. Numerous examples can be found in which pathogens express antigens that cross react with host antigens or induce local inflammatory responses that can lead to autoimmune responses through a very complex set of circumstances. Borrowing from the relationship between chronic infection with hepatitis C virus and autoimmune hepatitis as an example, we consider the possibility that infection with an unknown virus having specific tissue tropism could lead to a perceived autoimmune process. We raise the question whether such should be considered for Type 1 diabetes. PMID- 12848954 TI - Microchimerism in autoimmune disease: more questions than answers? AB - Recent studies indicate cell traffic occurs between the fetus and mother during pregnancy and that low numbers of fetal cells commonly persist in the maternal circulation for years thereafter. Microchimerism refers to a small number of cells or DNA from one individual harbored in another individual. Autoimmune diseases are more common among women and often increase in incidence following reproductive years. Chronic graft vs. host disease is an iatrogenic form of chimerism with similarities to some autoimmune diseases for which the HLA relationship of donor and host are of central importance. When considered together, these observations led to the hypothesis that microchimerism and HLA relationships of host and non-host cells are involved in autoimmune disease. The hypothesis is applicable to men, children and women without pregnancies because there are other sources of microchimerism, including from a twin, the mother or a blood transfusion. Microchimerism has now been investigated in a number of different diseases with some results supporting a potential role in disease pathogenesis. However, fetal and maternal microchimerism are also found in organs affected by non-autoimmune conditions. Moreover, microchimerism is commonly detected in the peripheral blood of healthy individuals raising the intriguing question of whether these cells are simple remnants of pregnancy or whether they might also have beneficial effects for the host. PMID- 12848955 TI - The antiphospholipid syndrome: multiple faces beyond the classical presentation. PMID- 12848956 TI - New insights into the pathogenesis of systemic sclerosis. AB - Systemic sclerosis (SSc) is a connective tissue disorder characterized by vascular abnormalities and excessive collagen synthesis. Extracellular matrix overproduction by fibroblasts results from abnormal interactions among endothelial cells, mononuclear cells (lymphocytes and monocytes) and fibroblasts, in a setting of vascular hyperreactivity and tissue hypoxia. Many autoantibodies have been identified in the sera of SSc patients; some of them are specific to the disease, such as anti-centromere antibodies in limited SSc, anti topoisomerase 1 and anti-RNA polymerase I/III antibodies in diffuse SSc. Their pathogenetic role(s) remains uncertain. However, genetic, environmental and possibly alloreactive factors might also contribute to disease susceptibility. PMID- 12848957 TI - Anti-neutrophil cytoplasmic antibodies and pathogenesis of small vessel vasculitides. AB - Anti-neutrophil cytoplasmic antibodies (ANCA) are a heterogenous group of autoantibodies with a broad spectrum of clinically associated diseases. ANCA testing has been established as a useful tool for the diagnosis of small vessel vasculitides, especially of 'ANCA-associated vasculitides' (AAV), such as Wegener's granulomatosis, microscopic polyangiitis and Churg-Strauss syndrome, in which circulating ANCA are commonly found. Within the last 20 years these antibodies were subject of intensive studies and a growing body of evidence arose for a distinct role of ANCA in the pathogenesis of the AAV. Our current concept of whether ANCA directly or indirectly contribute to vascular damage (ANCA cytokine-sequence-theory) was mainly developed from in vitro studies and is supported by data from clinical investigations as well as animal models. Recently a direct causal link between ANCA and the development of glomerulonephritis and vasculitis has been demonstrated. We now know that a passive transfer of ANCA is sufficient to induce disease, but it remains to be discovered how the autoantibodies to neutrophil antigens might triggered. PMID- 12848958 TI - From proteomics to molecular epidemiology: relevance of autoantibodies. PMID- 12848959 TI - Modifications of arginines and their role in autoimmunity. AB - Posttranslational modifications of proteins often perform a key role in the biological functioning of proteins. Some of these modifications also change the immunogenicity of proteins and peptides and create 'self'-antigens which might induce autoimmune responses. In particular modifications of arginines within a defined protein context can lead to a specific B-cell immune response. This review discusses the generation of such modifications and their relevance for autoimmunity. PMID- 12848960 TI - Autoantibody-induced apoptosis as a possible mechanism of autoimmune retinopathy. AB - Circulating antibodies specific to retinal proteins have been associated with retinal dysfunction in patients with retinopathy. Anti-recoverin antibodies found in patients with cancer-associated retinopathy (CAR) represent a unique model to study the relationship between retinal degeneration and autoimmunity. A body of evidence from in vitro and in vivo studies indicates that anti-recoverin autoantibodies are cytotoxic to retinal cells and induce apoptotic death of retinal photoreceptor cells, which leads to the degeneration of the photoreceptor cell layer. Similar to anti-recoverin autoantibodies, antibodies with other retinal specificities induce their target retinal cell death by activating a caspase 3-dependent apoptotic pathway. Thus, autoantibody-induced apoptosis may be a common pathway that leads to retinal death and blindness. PMID- 12848961 TI - Chronic urticaria and autoimmune thyroid disease: is there a link? AB - Chronic idiopathic urticaria (CIU) is a common and frustrating disorder. A subset of patients with CIU has been classified as autoimmune on the basis of two main findings: association with thyroid autoimmunity and anti-IgE and/or anti-IgE receptor antibodies. There is growing evidence that some cases of CIU are associated with thyroid autoimmunity. Patients with CIU have an increased frequency of Hashimoto thyroiditis with the presence of antibodies to thyroglobulin or a microsomal-derived antigen (peroxidase), even in euthyroid patients. No broad non-specific autoimmunity is present. Antibodies reactive with Fc(epsilon)RI, the high-affinity IgE receptor are found in sera of 10-40% of patients with CIU. A smaller subset of patients with CIU lack anti-Fc(epsilon)RI antibodies and have anti-IgE antibodies that cross-link Fc(epsilon)RIs occupied by IgE. There are a minority of CIU patients that lack such antibodies, but can degranulate basophils by a serum factor that has not yet been determined. Most patients with anti-IgE antibodies have an IgG antibody directed against the 34 kd alpha subunit of the IgE receptor. There are no data to suggest that any of the anti thyroid antibodies is pathogenic in terms of CIU, and most likely these are associated, parallel, autoimmune events. Screening for thyroid autoimmunity and function is advisable in all patients with CIU for the early identification of patients requiring either treatment of underlying thyroid dysfunction or follow up. PMID- 12848962 TI - The role of iron in T cell development and autoimmunity. AB - Iron is a vital metal for the proliferation of all cells including those of the immune system. Iron deficiency causes several defects in both the humoral and cellular arms of immunity. One of the most profound changes is a reduction in peripheral T cells and atrophy of the thymus. The presence of transferrin receptor on immature, proliferating thymocytes and the inhibition of thymocyte proliferation and differentiation by anti-transferrin receptor antibody highlight the importance of iron to T cell development. Growing evidence suggests that T cells may in turn, regulate iron metabolism perhaps through interactions with the non-classical major histocompatibility complex gene HFE. The association of the iron transporter NRAMP1 with several autoimmune disorders along with evidence that iron can catalyze the production of cryptic epitopes of several autoantigens, establishes a potential role for iron in the development of autoimmunity. PMID- 12848963 TI - 10th International Congress on Antiphospholipid Antibodies--summary. AB - The 10th International Congress on Antiphospholipid Antibodies (Sicily, Italy, September 29-October 3, 2002) (Fig. 1) provided enlightening aspects on the recent developments in antiphospholipid syndrome (APS) and antiphospholipid antibodies in more than 150 lectures and posters. Researchers from all aspects of medicine attended the meeting, implicating the systemic characteristics of APS. The important breakthroughs are summarized. PMID- 12848964 TI - Antiphospholipid syndrome: pathogenic mechanisms. AB - Despite the strong association between antiphospholipid antibodies (aPL) and thrombosis, the pathogenic role of aPL in the development of thrombosis has not been fully elucidated. Proposed pathophysiological mechanisms may be categorized into two types. First, aPL may act in vivo by disrupting the kinetics of the normal procoagulant and anticoagulant reactions occurring on cell membranes. Second, aPL may stimulate certain cells thereby altering the expression and secretion of various molecules. In this article, we review the mechanisms by which aPL may develop thrombotic events. PMID- 12848965 TI - Autoantibodies to ion channels at the neuromuscular junction. AB - Autoantibodies directed against voltage- or ligand-gated ion channels and their associated proteins at the neuromuscular junction give rise to a family of neurological autoimmune diseases. Antibodies to acetylcholine receptors or muscle specific kinase present on the postsynaptic muscle membrane are associated with different forms of myasthenia gravis (MG). Antibodies to the presynaptic voltage gated potassium and calcium channels are responsible for acquired neuromyotonia and Lambert-Eaton myasthenic syndrome (LEMS), respectively. The patients respond to immunotherapies and their plasma can transfer defects in neuromuscular transmission to mice, indicating that these are antibody-mediated conditions. In a small proportion of cases, ion channel antibodies have also been implicated in neurological dysfunction in the central nervous system. In these conditions, a proportion of the patients have an underlying tumour, thymoma in both MG and neuromyotonia and small cell lung carcinoma in LEMS, emphasising the putative role of autoimmunity to tumour antigens as a cause of neurological disease. PMID- 12848966 TI - B lymphocytes contribute to autoimmune disease pathogenesis: current trends and clinical implications. AB - Abnormal B lymphocytes influence the pathogenesis of many autoimmune diseases, in addition to serving as the origin of pathogenic autoantibodies. Although aberrant B cell function and autoimmunity have complex polygenic origins, recent studies in mouse models of autoimmune diseases have revealed overlapping defects in signal transduction pathways that alter B cell survival or activation, and lead to an autoimmune phenotype. Discovery of these important signaling pathways in mice has lead to an intense search for B cell abnormalities that correlate with autoimmune diseases in humans. This search has identified potential targets for therapeutic intervention that are the focus of planned and ongoing human clinical trials. This promises an arsenal of highly targeted, less toxic therapies focused on restoring normal B cell function that will eliminate pathogenic autoantibodies and replace the current use of immunosuppressive drugs. PMID- 12848967 TI - Antibodies against alpha-fodrin in Sjogren's syndrome. AB - Alpha-fodrin is a part of the membrane skeleton and expressed in the majority of mammalian cells. It is cleaved in apoptosis by caspase 3. One of the cleavage products, a 120-kDa protein, represents a neoantigen. Antibodies against that cleavage product of alpha-fodrin have originally been described in a murine model of Sjogren's syndrome. In addition, they are also present in up to 93% of patients with Sjogren's syndrome, depending on the stringency of the classification used. Although antibodies against alpha-fodrin are observed in other diseases characterized by chronic apoptosis, they are a valuable laboratory marker in the evaluation of Sjogren's syndrome. PMID- 12848968 TI - Liver transplantation for primary biliary cirrhosis. AB - Liver transplantation is now the accepted treatment for patients with end-stage disease or intractable symptoms. The success rate is high. Those grafted for PBC are at greater risk of developing acute and chronic rejection and are less likely to be weaned from immunosuppression. Following transplantation, AMA persist and histological features of PBC may be seen in the allograft, in up to 50% by 10 years. PMID- 12848969 TI - MHC class II peptide flanking residues of exogenous antigens influence recognition by autoreactive T cells. AB - Molecular mimicry between exogenous microbial antigens and self-epitopes has been proposed as a triggering mechanism for autoimmune diseases for many years. We reported that a peptide from a protein specific to Chlamydia pneumoniae (Cpn0483) which shares a motif with the dominant encephalitogenic epitope of the self antigen, rat myelin basic protein (rat68-86), elicits experimental autoimmune encephalomyelitis (EAE) in Lewis rats. We recently observed that rat68-86 utilizes aspartic acid (D) and arginine (R) in the common motif as primary and secondary TCR contacts, respectively. In contrast, the encephalitogenic activity of Cpn0483 is dependent on R and the C-terminal asparagine (N), which flanks the MHC class II-P9 anchor residue. Thus, rat68-86 and Cpn0483 share a common motif, are encephalitogenic and are both restricted by MHC class II RT1.B(l). T cells from rats immunized with the encephalitogenic Cpn0483 peptide proliferated to the priming peptide as well as to the non-encephalitogenic CpnN>A analog. However, CpnN>A-primed T cells did not respond to the native Cpn0483 peptide. We conclude that the MHC-flanking C-terminal asparagine residue markedly influences T cell recognition by the chlamydial peptide. PMID- 12848970 TI - Pathogenesis of Sjogren's syndrome. AB - The pathogenesis of Sjogren's syndrome is poorly understood. Genetic and environmental factors appear to contribute to the development of this syndrome. Viral infection is one of the most likely environmental factors. The primary lesion of Sjogren's syndrome is in the exocrine glands. A majority of the infiltrating cells in the lesion are CD(4+) CD45RO(+) memory T cells. Although antigen-presentation to T cells seems to occur in the exocrine tissues, these T cells are not fully activated. On the other hand, B cells comprise approximately 20% of the infiltrating cells, and several features of this syndrome are attributed to stimulated B cells. The presence of autoantibodies, such as anti-SS A/Ro and SS-B/La antibodies, is one of the characteristic features and is associated with severe disorders. Some antibodies appear to play a direct pathogenic role, for example, in cases of congenital heart block and sicca symptoms. Chronic inflammation with possible T cell-dependent antigen stimulation appears to induce neoplastic transformation of lymphocytes. PMID- 12848971 TI - T cell immunity and primary biliary cirrhosis. AB - T lymphocytes play a pivotal role in the autoimmune response in primary biliary cirrhosis (PBC). Recent studies have shown that there is overlapping in the PDC E2-specific T and B cell epitopes. In addition, helper T and cytotoxic T cell epitopes all contain a shared peptide sequence. In addition, recognition of exogenous antigens including bacterial antigens by autoantigen-specific T cell and the mechanism of molecular mimicry provide a clue to clarifying the pathogenesis of PBC. Furthermore, the findings that autoantigen-immune complexes cross present and also that the presentation of autoantigen is of a higher relative efficiency, define a unique role of autoantibodies in the pathogenesis of the autoimmune disease. The mechanism of immune-mediated bile duct damage in PBC, including the possible role of T cell-mediated cytotoxicity and molecular mimicry is discussed. PMID- 12848972 TI - Chemical-specific properties co-determine the type of adverse immune response. AB - Many drugs but also environmental pollutants may cause adverse reactions in susceptible individuals that are reminiscent of autoimmune syndromes. Apart from a number of predisposing often inherent, idiosyncratic determinants, chemical specific properties might be involved as well. Notably, reactive chemicals or metabolites may provoke formation or release of immunosensitizing neo-antigens (a.o. hapten-carrier complexes or cryptic epitopes). In addition reactive chemicals but also certain inert chemicals may trigger macrophages and other inflammatory cells to release proinflammatory products that, via elicitation of costimulatory help, support hapten- or neo-antigen-specific T cell activation. In addition, chemicals may influence immunoregulatory processes and modulate for instance the balance between type 1 and type 2 responses. Here, we review data showing that chemically induced upregulation of second or costimulatory signals co-determines not only whether, but also what type of an adverse immune response (type 1 or type 2) is triggered. PMID- 12848973 TI - Autoantibodies to factor VIII with catalytic activity. AB - Hemophilia A is an X-linked, recessive, bleeding disorder caused by defective or deficient factor VIII (FVIII) molecules. Infusion of purified FVIII to patients with severe hemophilia A results in approximately 25% of the cases, in the emergence of anti-FVIII antibodies (inhibitors) that are known to neutralize the pro-coagulant activity of FVIII by steric hindrance. We recently reported on the proteolysis of FVIII by allo-antibodies in the plasma of high responder patients with severe hemophilia A, demonstrating a new mechanism by which FVIII inhibitors may prevent the pro-coagulant function of FVIII. Hemophilia is the first model where a direct link between the hydrolysis of the target molecule and the occurrence of the clinical manifestations may be established. It also represents the first example in humans, of the induction of catalytic antibodies following the exogenous administration of an antigen. The characterization of FVIII inhibitors as site-specific proteases may provide new approaches to the treatment of inhibitors. PMID- 12848974 TI - Apoptosis and the liver: relation to autoimmunity and related conditions. AB - Apoptosis is a normal physiologic form of cell death that follows activation of either an intrinsic or extrinsic pathway. In the intrinsic, various stimuli, such as oxidative stress, lead to mitochondrial dysfunction and the release of pro apoptotic factors. Ligand binding to cell surface death receptors, such as Fas, activates the extrinsic pathway. Due to the rapid clearance of apoptotic cells, detection and quantification of apoptotic cells is prone to underestimation. In the liver, the importance of apoptosis is evident both during development and homeostasis of the biliary tree. Apoptosis also plays a prominent role in liver pathogenesis. Induction of the extrinsic apoptotic pathway by cytotoxic lymphocytes predominates in autoimmune liver diseases, viral hepatitis, and liver allograft rejection. Biliary cell apoptosis is highly regulated by bcl-2 family members. Both the extrinsic and intrinsic pathways are active in alcohol-related liver disease. Overexpression of anti-apoptotic proteins and FasL allow liver tumor cells to evade tumor specific cytotoxic lymphocytes. Agents that modulate apoptosis may be of future therapeutic benefit in a number of liver diseases. PMID- 12848975 TI - Application of tetramer technology in studies on autoimmune diseases. AB - Autoreactive T cells are thought to play a role in the immunopathogenesis of autoimmune diseases. Analysis of such cells had long been hampered by lack of suitable assays. Recently developed tetramer technology is based on the recognition of specific peptide-MHC complex by T cell receptor and on the increased binding affinity of multimerized peptide-MHC complex. MHC class I and class II tetramers can be used to detect autoreactive CD4(+) and CD8(+) T cells, while nonclassical MHC (such as CD1d) tetramer can be used to detect other T cell groups, for example natural killer T cells. Tetramer technologies enable direct quantitation of autoreactive T cells in blood and affected tissues. It is also possible to carry out phenotypic and functional characterization of specific T cells on a single cell basis by using tetramers. Of special interest, in situ tetramer staining has the great potential of analyzing autoreactive T cells in their cellular environments. Utilization of tetramers in studies of autoreactive T cells is expected to generate important information regarding the role of such cells in the underlying mechanisms of autoimmune diseases. PMID- 12848976 TI - Dosage and characterization of circulating DNA: present usage and possible applications in systemic autoimmune disorders. AB - The discovery of extracellular nucleic acids in the circulation was firstly reported in 1948. In the last few years it has been demonstrated that the entire spectrum of genetic changes seen in primary tumors could also be detected in the serum of patients with solid tumors. This observation has also opened up exciting possibilities for tumor detection and monitoring. More recently investigators started looking for other forms of non-host DNA in the plasma/serum so that in 1997 the presence of fetal DNA in the plasma/serum of pregnant women was demonstrated. This finding suggested that maternal plasma fetal DNA would be a very valuable material for noninvasive prenatal diagnosis and monitoring. It has been also postulated that the presence of the two-way trafficking of nucleated cells and free DNA between the mother and fetus may have potential implications for the development of certain autoimmune diseases. Concerning autoimmune disorders, Tan was the first author to describe the presence of high levels of circulating DNA in patients with systemic lupus erythematosus (SLE) in 1986. Later on different authors demonstrated that elevated levels of serum DNA was also present in patients with other diseases including rheumatoid arthritis. We have analyzed both circulating free DNA and DNA extracted from nucleated blood cells in scleroderma and in lupus patients but, by using gel electrophoresis, we were able to define the pattern of the DNA, instead of simply dosing its amount in the circulation. We have found that SLE and SSc have anomalous patterns of DNA both in serum and in the Buffy-coat and that these patterns are typical for each disorder. It is possible that understanding the biological significance of the diversity in DNA pattern exhibition in white blood cells may give new insights into the pathophysiology of autoimmune disorders. It is also conceivable that circulating and immune-competent cellular DNA markers might offer the promise of precise quantitative analysis useful for diagnostic purposes, without the need to establish difficult cutoffs as is necessary for protein markers. PMID- 12848977 TI - Epitope spreading and molecular mimicry as triggers of autoimmunity in the Theiler's virus-induced demyelinating disease model of multiple sclerosis. AB - The pathogenesis of multiple sclerosis (MS), a human demyelinating disease of the central nervous system (CNS), is currently unknown. It is widely thought that MS is an autoimmune disease which is supported by animal studies showing that myelin specific CD4+ T cells can induce similar clinical disease in mice as observed in MS. However, the mechanism(s) of activation of these autoreactive CD4+ T cells are unknown. Although genetic susceptibility is important, other factors may be involved. Viral infections have long thought to be involved in the pathogenesis of MS although there exists little or no direct evidence implicating a role for a specific virus in MS pathogenesis. This review will discuss two models of virus induced CNS autoimmunity, molecular mimicry and epitope spreading. These two mechanisms of activation of autoreactive T cells are presented in the context of MS. PMID- 12848978 TI - Rheumatic fever: the T cell response leading to autoimmune aggression in the heart. AB - Molecular mimicry between streptococcal and heart components has been proposed as the triggering factor leading to autoimmunity in rheumatic heart disease (RHD). CD4+ T cells apparently are the ultimate effectors of chronic heart lesions. In this review we summarize the studies on the T cell response of peripheral blood mononuclear cells and T cell clones infiltrating heart lesions from RHD patients against streptococcal M protein peptides and human heart tissue proteins. The T cell receptor usage and the cytokine profile of intralesional mononuclear cells are also presented. PMID- 12848979 TI - Autoreactive T cell responses in pemphigus and pemphigoid. AB - Pemphigus and pemphigoid are cutaneous autoimmune diseases characterised by autoantibodies directed against specific adhesion proteins of the epidermis and dermal-epidermal junction. These proteins are usually associated with desmosomes or hemidesmosomes. Binding of antibodies to their targets leads to the loss of cell-cell or cell-matrix adhesion and subsequently to blister formation. The humoral aspects of the autoimmune responses in pemphigus and pemphigoid have been extensively studied in the past. More recently, the cellular interactions resulting in the formation of autoantibodies and the involvement of autoreactive T cells in these diseases have attracted increased interest. In this review, the current knowledge on T cell involvement in pemphigus and pemphigoid is summarised. PMID- 12848980 TI - Improving the effectiveness of autoantibody testing in the clinic. AB - To enable clinicians to use autoantibody tests sensibly and economically, laboratories providing the test service have a responsibility to supply the test 'specifications'. This should include information about the test's validity and reliability, its normal range or range within a control population, its sensitivity and its specificity for different conditions against both control subjects and those with disorders which need to be distinguished by the test. This information will permit the development of predictive values, or better, of likelihood ratios which can then be used to generate clinically useful post-test probabilities. To date, autoantibody tests have been used to advantage by those skilled in their interpretation but often poorly by those less familiar with the area. It is appropriate now to demystify these tests by underpinning their clinical use with good operational data. PMID- 12848981 TI - CD8+ T suppressor cells are back to the game: are they players in autoimmunity? AB - The CD8+ T suppressor lymphocytes identified in humans belong to three different subpopulations. All of them inhibit the proliferation of antigen-specific T cells. The type 1 and type 2 of CD8+ T suppressor cells are characterized by the CD8+CD28- phenotype, while no detailed data are available at the moment on the phenotype of the type 3 of CD8+ T suppressor cells. The type 1 of CD8+ suppressor T lymphocytes acts by inducing alteration of expression of co-stimulatory molecules on dendritic cells. A cell-to-cell contact is required to mediate this effect. The type 2 of CD8+ T suppressor cells induces inhibition via cytokine secretion (IFNgamma, IL6) and do not need to interact directly with antigen presenting cells. The type 3 of CD8+ T suppressor cells mediates its function through the secretion of IL10. The complexity and multiplicity of CD8+ T suppressor cell subsets suggests that these cells may have an important role in the regulation of the immune homeostasis, acting together with the CD4+ T regulatory cell subpopulations. The specificity of the functions of each of these suppressor/regulatory subsets in the immune network requires to be clarified to better understand the immune system, its functions and the possibilities to modulate its activities in the course of immune-mediated diseases. PMID- 12848982 TI - Sex hormones and rheumatoid arthritis. AB - Sex hormones are implicated in the immune response, with estrogens as enhancers at least of the humoral immunity and androgens and progesterone as natural immune suppressors. In male rheumatoid arthritis (RA) patients, androgen replacement seems to ameliorate the disease and supports their involvement in the pathophysiology of the disease. The combination of androgens with cyclosporin A or methotrexate has been found to potentiate the apoptosis of monocytic inflammatory cells as well as to reduce the cell growth at least in vitro. Considerable interest has been devoted in the last years as to whether the use of oral contraceptive pills (OCs) may have a protective effect on the risk of RA. The results of many controlled studies have been found contradictory. At the present time, no consensus has been achieved regarding OCs administration and its relationship to the prevention or development of RA. In addition, an association of estrogen receptor gene polymorphism with age at onset of RA has been observed and might further explain inter-individual clinical and therapeutical-response variations. Local increased estrogen concentrations and decreased androgen levels have been observed in RA synovial fluids and seem to play a more important role in the immune/inflammatory local response. PMID- 12848983 TI - Immunopathogenesis, loss of T cell tolerance and genetics of autoimmune gastritis. AB - Over the past 10 years experimental autoimmune gastritis has been established as a highly defined model of organ-specific autoimmunity. Autoimmune gastritis represents one of the few autoimmune diseases in which the causative autoantigens, namely the gastric H/K ATPase alpha- and beta-subunits, are defined. Furthermore, it has been clearly established that a CD4+ T cell response to the H/K ATPase beta-subunit, in particular, is essential for the initiation of autoimmune gastritis. The immunopathology of autoimmune gastritis is due to a disruption of the normal developmental pathways of the mucosa, rather than a direct depletion of the end-stage parietal and zymogenic cells. CD4+CD25+ regulatory T cells were first described in experimental autoimmune gastritis and there has been a recent explosion of interest in the potential role of these immunoregulatory T cells in protection against a variety of autoimmune diseases. The availability of H/K ATPase deficient mice has begun to provide considerable insight into the basis for tolerance to the gastric autoantigens. Experimental autoimmune gastritis has also provided valuable insight into our understanding of the genetics of disease susceptibility and four distinct genetic regions have been identified which confer susceptibility to this organ-specific disease. The highlights of these recent advances are the subject of this review. PMID- 12848984 TI - Treatment of pregnancy loss in Hughes syndrome: a critical update. AB - Recurrent pregnancy loss is one of the main manifestations of antiphospholipid (Hughes) syndrome (APS). Fetal deaths (beyond the 10th week of gestation) are the most typical obstetric complication of APS. Data from controlled therapeutic trials are difficult to analyse, due to small samples and great heterogeneity in the obstetric history and antiphospholipid antibody profiles of women included. Corticosteroids are more harmful than beneficial. Heparin and aspirin are the drugs of choice for APS-related miscarriage, although it is not clear whether combination of both drugs is necessary for all women. The role of immunoglobulins is not well defined. PMID- 12848985 TI - Autoantibodies to heparan sulfate proteoglycans. PMID- 12848986 TI - Chemokines in autoimmunity: from pathology to therapeutics. AB - Leukocyte recruitment, accumulation, and activation have been both a unifying and enigmatic feature of a variety of autoimmune pathologies. While these processes were not well-known for decades, recent scientific discoveries have underscored the importance of specific chemokines in the evolution of autoimmunity. This has been supported by in vivo data from clinical studies and animal model experiments. Although chemokines are an attractive target for drug development, there are hurdles that need to be cleared. Nonetheless, the quest to understand chemokine biology and develop effective therapies continues to capture the imagination of scientists in academia and pharma alike. PMID- 12848987 TI - Anti-chromatin (anti-nucleosome) autoantibodies. AB - Anti-chromatin autoantibodies were one of the first autoantibodies ever detected since they make up the majority of antibodies causing LE Cell formation. Anti chromatin autoantibodies have had many names over the last few decades: LE cell factor; anti-nucleosome; anti-deoxyribonucleoprotein (DNP); and anti-(H2A-H2B DNA). These autoantibodies are found in approximately 75% of people with systemic lupus erythematosus and up to 100% of people with drug-induced lupus. They are also found in 20-50% of patients with autoimmune hepatitis type I (lupoid hepatitis). Anti-chromatin are not generally found in any other disease, thus showing very good sensitivity and specificity for patients with lupus, drug induced lupus and lupoid hepatitis. A number of studies have shown that in patients with lupus, anti-chromatin often correlates better with kidney disease than anti-DNA. Recent genetic analyses of murine models of lupus have identified at least three loci that work together to cause anti-chromatin antibodies and glomerulonephritis in mice. It will be an important breakthrough when the functions of the genes at these loci are identified. PMID- 12848988 TI - HIV and autoimmunity. AB - The association of immune dysfunction in patients with human immunodeficiency virus (HIV) infection and AIDS and the development of autoimmune diseases is intriguing. Yet, the spectrum of reported autoimmune phenomena in these patients is increasing. An infectious trigger for immune activation is one of the postulated mechanisms and derives from molecular mimicry. During frank loss of immunocompetence, autoimmune diseases that are predominantly T cell subtype CD8 driven predominate. There is evidence for B cell stimulation and many autoantibodies are reported in HIV patients. We propose a staging of autoimmune manifestations related to HIV/AIDS manifestations and the total CD4 count and viral load that may be beneficial in identifying the type of autoimmune disease and establishing the proper therapy. In stage I there is the acute HIV infection, and the immune system is intact. In this stage, autoimmune diseases may develop. Stage II describes the quiescent period without overt manifestations of AIDS. However, there is a declining CD4 count indicative of some immunosuppression. Autoimmune diseases are not found. During stage III there is immunosuppression with a low CD4 count and the development of AIDS. CD8 T cells predominant and diseases such as psoriasis and diffuse immune lymphocytic syndrome (similar to Sjogren's syndrome) may present or even be the initial manifestation of AIDS. Also during this stage no autoimmune diseases are found. In stage IV there is restoration of immune competence following highly active anti-retroviral therapy (HAART). In this setting, there is a resurgence of autoimmune diseases. The frequency of reported rheumatological syndromes in HIV-infected patients ranges from 1 to 60%. The list of reported autoimmune diseases in HIV/AIDS include systemic lupus erythematosus, anti-phospholipid syndrome, vasculitis, primary biliary cirrhosis, polymyosits, Graves' disease, and idiopathic thrombocytopenic purpura. Also, there is an array of autoantibodies reported in HIV/AIDS patients which include anti-cardiolipin, anti-beta2 GPI, anti-DNA, anti-small nuclear ribonucleoproteins (snRNP), anti-thyroglobulin, anti-thyroid peroxidase, anti myosin, and anti-erythropoietin antibodies. The association of autoantibodies in HIV-infected patients to clinical autoimmune disease is yet to be established. With the upsurge of HAART, the incidence of autoimmune diseases in HIV-infected patients is increasing. In this review, we describe the various autoimmune diseases that develop in HIV/AIDS patients through possible mechanisms related to immune activation. PMID- 12848989 TI - Accelerated atherogenesis in autoimmune rheumatic diseases. AB - The observation that systemic inflammatory rheumatic diseases such as rheumatoid arthritis (RA) are associated with a significantly increased rate of cardiovascular disease, which often occurs at a younger age than in the normal population, is particularly important given the increasing interest in the role of inflammation in atherogenesis in the general population. This review examines the accumulating evidence for accelerated atherogenesis of RA and updates the hypothesis that vasculitis plays a major role in this. Endothelial dysfunction (ECD), widely regarded as initial lesion in atherogenesis, has been shown to occur commonly in primary vasculitis. This ECD is a diffuse event, demonstrable in more than one vascular bed. It is not simply due to scarring in the vessel wall, related to the focal inflammation of the underlying vasculitis, since it may be reversed by suppression of the immune inflammation. However, the mechanisms for this ECD differ from that of the primary vasculitis. Preliminary evidence suggests that inflammatory mediators such as CRP, TNF, or sphingolipids may be involved. The diffuse ECD of vasculitis may have important consequences for both the progression of the primary disease and for cardiovascular events. A model for the role of vasculitis-induced ECD in the accelerated atherogenesis of rheumatic diseases is presented. These concepts are discussed together with the messages they suggest for 'idiopathic' atherosclerosis in the general population. PMID- 12848990 TI - Autoantibody-mediated atherosclerosis. AB - Beta2-glycoprotein I (beta2-GPI) is a major antigen for antiphospholipid antibodies (aPL) present in patients with antiphospholipid syndrome (APS). Oxidized low-density lipoprotein (oxLDL) is subsequently targeted by beta2-GPI and anti-beta2-GPI autoantibodies. Ligands specific for beta2-GPI derived from oxLDL have been characterized as oxidized forms of cholesteryl linoleate, such as 7-ketocholesterol-9-carboxynonanoate, i.e. 9-oxo-9-(7-ketocholest-5-en-3beta yloxy) nonanoic acid, (namely oxLig-1). The in vitro phenomenon that it is significantly increased in binding of oxLig-1 containing liposomes to macrophages via an interaction with beta2-GPI and an anti-beta2-GPI autoantibody (via the Fcgamma receptor) may propose a novel mechanism on 'autoantibody-mediated atherosclerosis'. Furthermore, autoantibodies against a complex of beta2-GPI and oxLig-1 are detected in sera of APS patients and appearance of the antibodies is associated with episodes of thrombosis, especially, arterial thrombosis. Thus, autoimmune atherogenesis linked to beta2-GPI interaction with oxLDL and autoantibodies may be present in APS. PMID- 12848991 TI - Pregnancy outcome in systemic lupus erythematosus: good news for the new millennium. AB - Systemic lupus erythematosus (SLE) is the autoimmune disease that most commonly compromises pregnancy. Moreover, the relationship between SLE and pregnancy is in both directions. However, in the recent years there has been a great change in the perception of the effects of pregnancy on SLE flares and of SLE on pregnancy outcome (both fetal and maternal). The current experience indicates that pregnancy in patients with SLE should not be regarded as an unacceptable high risk condition for the mother or her baby provided that careful planning of conception and multidisciplinary monitoring and treatment are carried out. PMID- 12848992 TI - Prolactin and autoimmunity. AB - Prolactin (PRL) is a versatile hormone that is produced by the anterior pituitary gland and various extrapituitary sites including immune cells. Furthermore, PRL has widespread influences on proliferation and differentiation of a variety of cells in the immune system and is, in effect, a cytokine. PRL-receptors (PRL-R) are distributed throughout the immune system and are included as members of the cytokine receptor superfamily. PRL-R signal transduction is mediated by a complex array of signaling molecules of which JAK2, Stat1 and Stat5 pathway have been well studied. In PRL-stimulated T cells, the transcription factor gene, interferon regulatory factor-1 provides a mechanism whereby PRL can regulate the immune response. The human PRL gene is situated on the short arm of chromosome 6 close to the major histocompatibility complex. Polymorphisms of the human PRL gene have implications for production of lymphocyte PRL in SLE. Mild and moderate hyperprolactinemia (HPRL) has been demonstrated in 20-30% of SLE patients and is associated with active disease. HPRL may have a role in lupus nephritis and central nervous system involvement of SLE patients. HPRL stimulated the production of autoantibodies. These evidences support the important role of PRL in autoimmunity and autoimmune diseases, mainly SLE. PMID- 12848993 TI - Antiendothelial cell antibodies in systemic lupus erythematosus. AB - Sera from patients with systemic lupus erythematosus have been reported to contain IgM and/or IgG binding to endothelial cells (EC), i.e. anti-EC antibodies (AECA). Similar autoantibodies have been claimed to occur in a number of conditions associated with vasculitis. The original cyto-enzyme-linked immunosorbent assay (ELISA) remains the most widely used method for the detection of AECAs, although numerous pitfalls have been identified since then. These difficulties may explain why a consensus on the prevalence of AECAs has not been reached thus far. It is therefore desirable to confirm a positive result in the cyto-ELISA using other methods, such as flow cytometry, immunoprecipitation or Western blot. Yet, these methods appear to be difficult to use on a routine basis. With regard to the AECA effects, their binding induces activation of ECs, as substantiated by up-regulation of adhesion molecules, and synthesis of cytokines and chemokines, followed by their secretion. Some of these autoantibodies encourage the local production of tissue factor, and thereby favour coagulation. Other AECAs trigger apoptosis of ECs, although the Fas receptor does not seem to be involved in this process. In fact, since the target antigens are not well defined, the current challenge is to identify EC target molecules, and thus to gain further insights into the pathogenesis of diseases with vasculitis. PMID- 12848994 TI - Heparin-induced thrombocytopenia with thrombotic sequelae: a review. AB - Heparin-induced thrombocytopenia (HIT) occurs in 1-5% of patients treated with heparin. The pathogenesis involves the formation of antibodies to heparin platelet factor 4 complexes, and the major clinical sequelae are thrombotic. Diagnosis is based on a combination of clinical and laboratory data. Treatment consists of stopping heparin, but, insofar as the risk of thrombosis remains high, treatment by alternative antithrombotic agents is indicated. Most clinical experience has been with danaparoid sodium and hirudin. The use of low-molecular weight heparins (LMWH) in subsequent HIT episodes has been described, but is not recommended, especially with the introduction of new agents, such as oral thrombin inhibitors and pentasaccharides, which are hoped to reduce the use of heparins and the occurrence of HIT. PMID- 12848995 TI - About CD4pos CD25pos regulatory cells. AB - Autoreactive cells that escape from thymic negative selection are either anergized or counter-regulated in the peripheral lymphoid organs by regulatory T (Treg) cells. Most investigated Treg cells are CD4pos cells expressing the alpha chain of the IL-2 receptor (CD25). Absence of this particular population leads to the development of a wide range of autoimmune disorders, suggesting a critical role for CD4pos CD25pos cells in the maintenance of normal immune homeostasis. Numerous studies have aimed at unraveling the mechanisms of regulation, focusing mainly on naturally occurring Treg cells (generated in naive animals) and regulation of CD4pos autoreactive cells. In contrast, generation following antigenic priming of CD4pos CD25pos Treg cells with non-self (viral) antigen specificity and regulation of virus-activated CD4pos and CD8pos cells remain to be investigated. In this review, we describe the data collected over the past few years of intense focus on CD4pos CD25pos Treg cells and present our approach to investigate the generation of antigen-specific Treg cells during the course of an anti-viral immune response. PMID- 12848996 TI - Genetic dissection of familial autoimmune thyroid diseases using whole genome screening. AB - The autoimmune thyroid diseases (AITDs) are multifactorial disease which are caused by genetic susceptibility and environmental triggers. Various epidemiological and genetic techniques can be employed to study the genetic contribution to disease development. Most epidemiologic data support an important genetic contribution to the development of AITD. The genetic susceptibility to AITD involves several genes with varying effects. Some AITD susceptibility genes are most likely immune modifying genes which increase the susceptibility to autoimmunity in general (e.g. HLA, CTLA-4) while others may be thyroid-specific (e.g. thyroglobulin). These genes probably act in concert to increase the autoimmune reactions in susceptible individuals and direct them towards the thyroid. PMID- 12848997 TI - Th2-type immunopathological manifestations induced by mercury chloride or gold salts in the rat: signal transduction pathways, cellular mechanisms and genetic control. AB - Heavy metals induce various immunopathological disorders including an increase in serum IgE concentration in predisposed humans. The effects of HgCl2 or gold salts differ depending on the strain of rats tested: they induce Th2-mediated immunopathology in Brown-Norway (BN) rats while HgCl2 triggers an immunosuppression in Lewis (LEW) rats. The disease is due to the emergence of self-MHC class II reactive Th2 cells in BN rats. Autoreactive T cells are also found in HgCl2-injected LEW rats but they produce TGFbeta and IL-10 and have immunoregulatory properties. Hg or Au act on the early steps of T cell activation resulting in IL-4 and IFNgamma gene expression with preferential IL-4 expression in BN rats. Analyzing the effects of HgCl2 on T cells led us to identify a new signaling pathway implicated in IL-4 production. An important feature of this model concerns genetics. Indeed Th2-dependent autoimmunity induced by metals occurs only in BN rats that are genetically committed to develop Th2 responses. Cellular features at play are discussed as well as the identification of loci that control both the Th1/Th2 balance and susceptibility to autoimmunity. PMID- 12848998 TI - Adoptive cellular gene therapy of autoimmune disease. AB - Autoimmune disorders represent inappropriate immune responses directed at self tissue. Because CD4+ T cells are important mediators in the pathogenesis of autoimmune disease, they are ideal candidates for cell-based gene therapy. Using retrovirally-transduced cells and luciferase bioluminescence, we have demonstrated that primary T cells and hybridomas, rapidly and preferentially home to the sites of inflammation in organ-specific autoimmune disease. These cells, transduced with retroviral vectors to drive expression of various 'regulatory proteins', such as IL-4, IL-10 and IL-12p40, deliver these immunoregulatory proteins to the inflamed lesions, providing therapy for experimental models of autoimmune disease such as EAE, CIA and NOD mice. This technique was originally developed in our lab in the murine model of multiple sclerosis, EAE, where T cell hybridomas reactive with myelin basic protein (MBP) were transduced to express and used to deliver the modulatory cytokine, IL-4. Recently we have observed that the cytokine receptor antagonist, IL-12p40 transduced anti-myelin basic protein (MBP) TCR-transgenic T cells (but not CII-reactive T cells) were effective in preventing EAE whereas the CII-reactive, but not MBP-reactive T cells, transduced to express IL-12p40, would treat CIA. PMID- 12848999 TI - Primary biliary cirrhosis and hepatocellular carcinoma. AB - The findings by epidemiological studies on the link between PBC and HCC are in general agreement with the notion that cirrhosis is a risk factor for HCC development. From the clinical perspective, this implies that in PBC patients with cirrhosis, the screening for HCC should be considered for evaluating prognosis as well as therapeutic options. At this time, it is not possible to determine whether any PBC-specific risk factors other than cirrhosis per se exist for the development of HCC. Identification of such risk factors may point to new mechanisms involved in the carcinogenesis of HCC. In order to answer the question whether the underlying mechanisms for PBC are risk factors for HCC, more aggressive clinical studies with larger patient populations are needed. Such studies should include patients with PBC as well as patients with cirrhosis of other etiologies, both have to be carefully matched for patient characteristics including race, gender, age, disease stage and period of follow-up. On the other hand, the resolution of this issue also relies on a better understanding of the molecular pathogenesis of PBC itself. PMID- 12849000 TI - The pathogenesis of inflammatory muscle diseases: on the cutting edge among the environment, the genetic background, the immune response and the dysregulation of apoptosis. AB - Inflammatory muscle diseases (IMD), including dermatomyositis (DM) and polymyositis (PM), affect skeletal muscle, leading to profound tissue modification. The etiology of IMD is unknown, but multiple steps of the disease pathogenesis have been identified. The main alterations involve the immune response. Cellular infiltrates found in the muscle provide strong evidence for the involvement of a preferential immune mechanism of muscle damage. The pathologic differences found between PM and DM indicate a different role played by cell-mediated and humoral immune alterations. It is well accepted that in the pathogenetic pathway both host genes and environmental factors are involved. Apoptosis, or programmed cell death, is a complex process that plays a key role in many physiological events. It regulates the turnover of immune cells and is one of the mechanisms involved in ensuring a competent, non-autoreactive repertoire of lymphocytes. Apoptosis as a mechanism of muscle fibre death has been described in several neuromuscular disorders and muscular dystrophies, and evidence of a lack of apoptosis in IMD suggests a failure of apoptotic clearance of inflammatory cells playing a role in the maintenance of chronic cytotoxic muscle fibre damage. Most likely, the failure of apoptosis seems to be the main hallmark of the pathogenesis of IMD. PMID- 12849001 TI - Autoimmunity, oxidized LDL and cardiovascular disease. AB - Atherosclerosis is the major cause of cardiovascular disease (CVD) and in addition to established risk factors as smoking, hypertension, diabetes and dyslipidemia, inflammation and autoimmune reactions have been much discussed recently. Several lines of evidence indicate that also inflammation and autoimmune reactions are highly relevant in atherosclerosis and CVD. Inflammatory cells and cytokines are present in lesions, already at an early stage; animal experiments suggest that immune reactions, though not necessary for development of atherosclerosis, can modulate disease development and systemic inflammation is associated with an enhanced risk of CVD. The enhanced risk of CVD in a major autoimmune disease, systemic lupus erythematosus (SLE), is therefore highly relevant, and in addition to being an important clinical problem, SLE-related CVD could give insights into the nature of autoimmunity in atherosclerosis and CVD in general. We recently defined traditional and non-traditional risk factors for CVD in SLE. These include increased atherosclerosis (as determined by intima-media thickness of carotid artery); raised oxidized low density lipoprotein (OxLDL) and autoantibodies to OxLDL; dyslipidemia with raised triglycerides and Lp(a) and decreased HDL-cholesterol concentrations; raised systemic inflammation; presence of anti-phospholipid antibodies including lupus anticoagulant, homocysteine levels and more frequent osteoporosis. Disease duration, smoking, blood pressure or diabetes mellitus did not differ significantly between the groups. Taken together, immune reactions are highly relevant in atherosclerosis, and patients with autoimmune disease like SLE are at high-risk of CVD. If confirmed prospectively, non-traditional risk factors like OxLDL in the circulation, autoantibodies against OxLDL and phospholipids and inflammation could lead to new therapeutic strategies and insight into disease mechanisms. PMID- 12849002 TI - Viral etiopathogenesis of Sjogren's syndrome: role of the hepatitis C virus. AB - Patients with hepatitis C virus (HCV) chronic infection present some extrahepatic manifestations that may mimic the clinical, immunologic and histological manifestations of primary Sjogren's syndrome (SS). Thus, HCV patients with sicca symptomatology and positive autoantibodies could be misdiagnosed as a 'primary' SS. Nevertheless, there are several clinical and immunologic features that could help us differentiate both processes. PMID- 12849003 TI - Autologous hematopoietic stem cell transplantation for Crohn's disease. AB - The feasibility and early experience of hematopoietic stem cell transplantation for severe Crohn's disease (CD) are discussed. The natural history, therapy, and evidence for autoimmunity of CD are outlined and related to this new therapy in terms of morbidity and mortality. PMID- 12849004 TI - Immunglobulin repertoire analysis provides new insights into the immunopathogenesis of Sjogren's syndrome. AB - This review focuses on the use of immunglobulin (Ig) variable region genes by B cells from patients with primary Sjogren's syndrome (pSS) and the biologic insights that this provides. Comparison of the Ig repertoire from the blood and parotid gland of pSS patients with that of normal donors suggests that there are typical disturbances of B cell homeostasis with depletion of memory B cells from the peripheral blood and accumulation/retention of these antigen-experienced B cells in the inflamed tissue. Although there are clonally expanded B cells in the parotid gland, generalized abnormalities in the B cell repertoire are also found in pSS patients. The vast majority of the current data indicate that there is no major molecular abnormality in generating the IgV chain repertoire in patients with pSS. In contrast, disordered selection leads to considerable differences in the V(L) gene usage and V(H) CDR3 length of the B cell Ig repertoire in pSS patients. The nature of the influences that lead to disordered selection in pSS remains to be determined, but should provide important clues to the etiology of this autoimmune inflammatory disorder. PMID- 12849005 TI - Pathogenesis of heparin-induced thrombocytopenia and thrombosis. AB - Heparin-induced thrombocytopenia and thrombosis (HIT/T) is a common immune mediated disorder often manifested by life-threatening thrombosis. There is increasing evidence to indicate that HIT/T is caused by antibodies to complexes between platelet factor 4 (PF4) and heparin that activate platelets, monocytes and vascular endothelium leading to the generation of thrombin. Advances in defining the immunological basis of HIT/T have yielded insights into the antigenic determinants, antibody-antigen interactions and effector responses that contribute to its pathogenesis. However, these studies also reveal that anti PF4/heparin antibodies develop far more commonly than clinically overt disease, raising questions as to serologic and other factors that predispose to clinical thrombocytopenia and thrombosis. An improved understanding of the natural history of HIT/T and the introduction of alternative anticoagulants have led to a somewhat improved clinical outcome. The recent development of a monoclonal anti heparin/PF4 antibody and the establishment of a murine model of HIT/T may help to better define the pathogenesis and management of this common autoimmune disorder. PMID- 12849006 TI - Hematopoietic stem cell therapy for type 1 diabetes: induction of tolerance and islet cell neogenesis. AB - Diabetes is a chronic disease with significant morbidity and mortality. Pancreas or islet cell transplantation is limited by a shortage of donors and chronic immune suppression to prevent allograft rejection. Consequently, interest exists in islet cell neogenesis from embryonic or mesenchymal stem cell as a possible cure for diabetes. However, unless tolerance to islet cells is re-established, diabetes treated by islet cell transplantation would remain a chronic disease secondary to immune suppression related morbidity. If islet cell tolerance could be re-induced, a major clinical hurdle to curing diabetes by islet cell neogenesis may be overcome. Recent studies suggest that adult hematopoietic stem cells (HSC) can reintroduce tolerance to auto-antigens. It is possible that HSC may also be able to switch lineage and, therefore, be a convenient source of stem cells for both inducing tolerance and islet cell regeneration. PMID- 12849007 TI - Insulin autoimmunity: prediction/precipitation/prevention type 1A diabetes. AB - Type 1 diabetes of both the NOD mouse and man is associated with autoimmunity directed against insulin which is the only beta cell specific autoantigen identified to date. One can use autoantibodies to insulin to predict diabetes, use insulin peptides to create insulin autoantibodies, insulitis and diabetes, and use insulin or its peptides in animal models to prevent diabetes. An expanding set of resources are now available for the development and testing in man of therapies to prevent type 1 diabetes, and a number of trials utilizing insulin peptides are now underway. PMID- 12849008 TI - Autoantibodies to IGF-II mRNA binding protein p62 and overexpression of p62 in human hepatocellular carcinoma. AB - Studies of autoantibodies in systemic rheumatic diseases have provided abundant evidence suggesting that autoimmune responses are antigen-driven and that autoantibodies often can be viewed as reporters of the immune system revealing the identity of antigens which might be playing roles in the pathophysiology of the disease process. Recent data from our laboratory suggest a similar mechanistic process may be involved in humoral immune responses in certain cancers such as hepatocellular carcinoma (HCC). HCC is unique in that one can follow a cohort of patients with chronic liver disease who will likely progress to develop malignancy over a period of 10 or more years. It has been observed that during transition from chronic liver disease to HCC, novel autoantibodies can appear which are not detected prior to pre-malignant conditions. The hypothesis is that these novel antibody responses may be stimulated by cellular proteins which are involved in carcinogenesis. By immunoscreening an expression library to isolate cDNA clones of autoantigens, a RNA-binding autoantigen p62 has been identified in HCC recently and autoantibodies to p62 were found in 21% of a cohort of HCC patients. p62 is a cytoplasmic protein which binds to mRNA encoding insulin-like growth factor II (IGF-II), a growth factor which is known to be overexpressed in HCC and is tumorigenic in transgenic animals. The expression of p62 is developmentally regulated, and expressed in fetal, but not in adult liver. Our recent observations showing that p62 was aberrantly expressed in 30% of unselected HCC suggest that it could play a role in HCC and other tumors by upregulating expression of growth factor IGF-II in the milieu of other oncogenic factors. PMID- 12849009 TI - The clearance of apoptotic cells: implications for autoimmunity. AB - Apoptosis has been clearly characterised by the ability to limit the activation of inflammatory responses through the disposal of the apoptotic cell by rapid uptake by phagocytes. The exposure of phosphatidylserine deriving from the loss of plasma lipid asymmetry is the early membrane signal which alerts the phagocyte about the imminent apoptotic death of the cell. Also modifications of membrane carbohydrate groups on apoptotic cells contribute to phagocyte recognition. Soluble proteins such as C1q, mannose-binding lectin, surfactant proteins A and D, C-reactive protein, C3bi, beta2-glycoprotein I and growth arrest specific gene 6 bind to apoptotic cells and act as "opsonins" thus favouring their clearance. A redundant and promiscuous system of receptors including integrins, scavenger receptors, CR3 and CR4, calreticulin, CD14 and Mer receptor ensures an efficient and rapid uptake of apoptotic cells. In animal models and in human pathology, single genetic defects of molecules involved in apoptotic cell clearance seem to be the main determinant in the development of autoimmunity. The uptake of apoptotic cells by phagocytes provides an immunomodulatory effect in that it triggers the release of anti-inflammatory cytokines, inhibits the production of inflammatory cytokines and leads to T cell tolerance. Impaired clearance of apoptotic cells or the presence of 'danger' signals may modify the balance between tolerance induction and activation of T cells leading to an effective autoimmune response. PMID- 12849010 TI - Sex ratio and rheumatic disease. AB - Some thyroid, rheumatic and hepatic diseases consistently have high female:male ratios, but many autoimmune diseases do not. Gonadal hormones, if they play a role in determining sex ratios, likely do so through a threshold or permissive mechanism. Sex differences related to X-inactivation, imprinting, X or Y chromosome genetic modulators, and intrauterine influences, exposures, vulnerable periods, or thresholds are alternative, theoretical, explanations for sex differences of incidence. PMID- 12849011 TI - The targets of nephritogenic antibodies in systemic autoimmune disorders. AB - In situ formation of immune complexes is a well recognized mechanism of renal injury in systemic autoimmune disorders. The identification of intrinsic renal antigens that are targets of nephritogenic antibodies is a field of active investigation. Recently, two proteins expressed in the kidney have been characterized as renal antigens. Alpha-actinin, an actin-binding protein localized in glomerular podocytes, is the major target of nephritogenic anti-DNA antibodies. Alpha-enolase, a glycolytic enzyme, is a target of nephritogenic anti DNA and non-anti-DNA antibodies. PMID- 12849012 TI - Role of profibrogenic cytokines secreted by T cells in fibrotic processes in scleroderma. AB - IL-4 is potentially a major profibrogenic cytokine upregulating the expression of collagen genes. In vivo studies have shown that the disruption of STAT6, IL-4R, IL-4 or transforming growth factor-beta (TGF-beta) genes in TSK mice, which develop scleroderma-like syndrome, prevented the occurrence of skin sclerosis and of autoantibodies. Additionally, it is known that the homozygosity of TSK mutation is lethal and the embryos die by day 7-8 of pregnancy. The disruption of IL-4 gene rescued from death TSK/TSK mice suggesting a role for IL-4 in embryonic development. Since TGF-beta should compensate the lack of IL-4 on regulation of collagen gene expression, we have studied the effect of IL-4 on the expression of TGF-beta gene. Our results showed IL-4 dependence of the transcription of TGF beta gene and that in both TSK/+ and TSK/TSK IL-4-/- mice the expression of TGF beta gene is impaired. PMID- 12849013 TI - What do we mean when we talk about ethical cancer care? PMID- 12849014 TI - Oncology patients' and oncology nurses' perceptions of nurse caring behaviours. AB - Previous research on patients' and nurses' perceptions of nurse caring behaviours has documented significant differences in the ranking of important behaviours. However, these samples have included a variety of medical-surgical patients and nurses and different types of institutional settings, all of which may have affected the results. The present study sought to determine if patients and nurses from one subspecialty area and one institution would have more concordant perceptions of caring. Forty-four oncology patients and 49 oncology nurses completed the Respondents Perceptions of Caring Behaviour Scale (RPCBS). Results showed that overall mean patient rankings were highly correlated with mean nurse rankings (Spearman's correlations coefficient 0.94, P<0.0001). The Wilcoxon two sample rank sum test was used to test the difference in rank of the 20 items between the patients and nurses. There was a significant difference in rank in only six of the 20 items. These data suggest that oncology patients and nurses have more concordant perceptions of caring than previously investigated groups. Implications for practice and further research are discussed. PMID- 12849017 TI - The experiences of patients and nurses with a nurse-led peripherally inserted central venous catheter line service. AB - Nurse-led Peripherally Inserted Central Venous Catheter (PICC) services are becoming commonplace in cancer centres across the UK. Research has shown that these services are cost-effective and are associated with fewer catheter-related complications, including infection and thrombosis, than skin-tunnelled catheters. This exploratory study aimed to explore patients' and nurses' experiences of a nurse-led PICC line service. Recorded interviews were conducted with 10 cancer patients with PICC lines inserted, five hospital-based nurses and five community based nurses experienced in caring for patients with these lines. Respondents' recorded accounts were transcribed, coded and analysed to determine patients' and nurses' perceptions of the service and the extent to which collaborative practice was promoted. Findings were synthesized into five themes: Education, Formation of Expectations, Confidence, Sensory Experience and Adaptation. All respondent groups were positive about the PICC line service and the specialist oncology nurses working within it. The findings did identify some aspects of the service which could be improved, including the timing of information given to patients and the use of more innovative educational approaches. This study has informed the development of a potential flexible service structure which may be incorporated into established and differing care settings using existing resources. PMID- 12849018 TI - The impact of expanding the numbers of clinical nurse specialists in cancer care: a United Kingdom case study. AB - In the last few years there has been an increase in the number of clinical nurse specialists (CNSs) working in cancer care throughout the UK. This paper examines the issues raised by the proliferation of site-specific cancer CNS posts, developed to give a 'nursing lead' for a particular tumour type. The issues common to all CNS posts are exacerbated when a number of specialists are involved in the cancer journey, causing potential fragmentation and confusion. Key issues need to be addressed if the role is to remain credible, and ensure clarity for patients and the multiprofessional team. These include role clarification, sensitive integration, interface with other cancer CNSs and the multiprofessional team, evaluation, cost, education and training, recruitment and succession planning and the 'career cul-de-sac'. Cancer nursing's response to these challenges will set the professional agenda for education, training and future models of service delivery. PMID- 12849019 TI - The development of a conceptual framework for advancing cancer nursing practice in Europe. AB - Continued professional development through formal education programmes is essential for improving the quality of care provided to patients with cancer. This paper describes a European Oncology Nursing Society project concerned with advanced cancer nursing education. While the development of an education programme for advanced cancer nursing practice in Europe proves difficult at this time, this project has provided the opportunity to examine some of the key elements of advanced cancer nursing work. In turn, the information gained has enabled the development of a conceptual framework for advancing cancer nursing practice in Europe to be constructed. PMID- 12849020 TI - Measurement of cancer attitudes: a review. AB - Many psychologists maintain that the attitudes we hold about a particular topic or subject will influence the way in which we behave towards it: positive attitudes lead to positive behaviours and vice versa. A literature search was conducted using the databases of MEDLINE and CINAHL. This produced research-based evidence showing that despite treatment advances, cancer remains one of the most feared of all diseases, not only among the public but also among oncology health care professionals. However, the practice of measuring attitudes is fraught with difficulties. This review article will discuss the link between attitudes and behaviour and review pertinent literature regarding public and professional attitudes towards cancer before critically analysing the most common tools for evaluating cancer attitudes and making recommendations for the future of attitude measurement. PMID- 12849021 TI - Evaluating the efficacy of scalp cooling using the Penguin cold cap system to reduce alopecia in patients undergoing chemotherapy for breast cancer. PMID- 12849022 TI - The association of fatigue with febrile neutropenia in patients receiving radiotherapy. PMID- 12849023 TI - Nursing management of chemotherapy-induced nausea and vomiting in children. Current prescribing and administration practice--is it being used to its full potential? PMID- 12849025 TI - Communication between nurses and simulated patients with cancer: evaluation of a communication training programme. AB - In this paper the effect of a communication training programme on the instrumental and affective communication skills employed by ward nurses during the admittance interview with recently diagnosed cancer patients was investigated. The training focused on teaching nurses skills to discuss and handle patient emotions. For this purpose, 46 nurses participated in 92 videotaped admittance interviews with simulated patients. The study had a randomized pre-test-post-test design. Multi-level analysis was used to measure the effects of the training. The results revealed that the trained nurses significantly increased asking open-ended psychosocial questions, which indicates that they were actively exploring patients' feelings. Furthermore, the patients showed a significant increase in affective communication. In conclusion, the results of this study demonstrate that, although limited, training can induce favourable changes in the communication skills of nurses, and can even affect patient communication. Future studies should focus on the further evaluation of educational programmes to enhance communication skills. PMID- 12849028 TI - Barriers to research utilization: the clinical setting and nurses themselves. AB - The advance of the evidence-based practice (EBP) movement has been evident in almost every Western country and health system over the past two or three decades, fueled by an ever-rising demand on resources. Nurses at all levels are increasingly expected to address the key challenge of EBP, which is to use research evidence in a conscientious, explicit and judicious way when making decisions about patient care. The main aim of the paper is to encourage nurses to embrace the challenge of EBP. First, as background, this paper presents key findings from the limited body of research which has examined barriers to research utilization in the nursing context. Nurses generally feel there are many barriers, with primary barriers being lack of time, lack of relevant skills, poor team-working and several aspects of nursing 'culture' (ritualistic care, no authority and no incentives). Some conceptual models in implementation of research findings are introduced, and a summary presented of key areas which nurses need to address when considering research utilization. PMID- 12849029 TI - Nursing in Colorectal Cancer Initiative: the audit phase. Part 2. Content validity of the audit tool and implications of the standards set for clinical practice. AB - This paper gives an account of the process of refining the content validity of an audit tool, which defines and measures best practice in colorectal cancer nursing and identifies areas for development. The European Oncology Nursing Society (EONS) and AstraZeneca collaborated to develop the Nursing in Colorectal Cancer Initiative (NICCI). The initiative was funded through educational grants from AstraZeneca and led by EONS. It is a project with two components, education and audit. The education component culminated in a manual that provides a core set of materials concerning fundamental aspects of colorectal cancer, to foster a common understanding amongst nurses at national and international levels. An audit tool was developed to measure standards of nursing care in relation to the delivery of cytotoxic chemotherapy to patients with advanced colorectal cancer. The content validity of the audit tool was established in three stages by expert panel review with revisions made to the content and organisation of the audit measures at each stage. The standards set by the NICCI Audit Project have key implications for multi-professional practice in colorectal cancer care. PMID- 12849030 TI - Clinically effective cancer care: working with families. AB - Families of those with cancer frequently report that their information and support needs are not being met to their satisfaction. Moreover there is evidence of prolonged stress and overt psychological morbidity within this group. Family nursing is suggested as a means by which nurses can assist families adapt to the stresses of supporting others with cancer, yet this has not been tested to any great extent. This paper offers a critical review of a selection of key studies where nursing has been shown to demonstrate positive health outcomes for families when a member has cancer. PMID- 12849031 TI - Using action research in paediatric oncology to develop an oral care algorithm. AB - The concurrent use of an oral assessment guide and oral care algorithm proved valuable in providing a more consistent approach to the oral care of children on a particular paediatric oncology ward (Gibson et al. 1997). However, as action research was chosen to develop practice, the diagnosis of new problems was anticipated. The identification of problem areas arose from two sources-informal feedback from the practice setting and via a formal audit using a tripartite approach consisting of a structured interview, vignettes and an analysis of existing documentation. The findings firstly demonstrated that practitioners experienced problems associated with the interpretation of the information contained within the existing algorithm. Secondly, there was also strong evidence to show that the production of a second 'therapeutic' algorithm would be beneficial to patient care. Finally, to ensure clarity, minor changes were required to the oral assessment guide. By using a collaborative approach between researchers and practitioners a positive resolution to a commonly identified clinical problem was expedited. PMID- 12849033 TI - The arts in cancer care: learning from experience. PMID- 12849032 TI - Palliative care in the community for children with cancer in South East England. AB - In-depth interviews and discussions were held with 40 different professionals in South East England involved in managing palliative care for children with cancer in the community. Participants included paediatric oncologists and outreach nurse specialists in tertiary centres, paediatricians in shared care units, children's community nurses, general practitioners, social workers and child psychologists. The research examined palliative care services available in the region, exploring attitudes to both current provision and possible service improvements. Providing palliative care in the community involves multi-agency collaboration and the study highlighted a range of different approaches to case management with the diversity of resources available. Key improvements proposed by health-care professionals included: better communication and liaison between all the professionals involved; clearer allocation of roles and responsibilities; 24-hour availability of specialist advice on palliative care for children with cancer; faster access to social work and psychology services at the community level; continuity of nursing and respite care. The provision of specialist local palliative care services for children with cancer was generally rejected. The participants favoured improving community palliative care for all children with life-limiting or life-threatening conditions with community nursing teams providing continuity of care and outreach nurses providing specialist advice and support. PMID- 12849034 TI - Stories about breast cancer in Australian women's magazines: information sources for risk, early detection and treatment. AB - Sixty articles in five Australian women's magazines were analyzed for journalistic qualities, metaphors, narrative features and accuracy of clinical facts related to risk, early detection and treatment of breast cancer. The stories were features, news features or soft news stories. The stories reflected the 'good news' editorial style of women's magazines. A dominant theme in the stories was that early detection of breast cancer is crucial and equals survival. While there were few inaccuracies in the stories, there was little detail of treatment modalities, an emphasis on lifestyle as a risk factor and a prevailing message that a genetic history of breast cancer means you will get it. A major implication of the findings is that nurses, who provide information to women, must be aware of the goals of journalists and the educational power of narrative logic of stories in women's magazines. PMID- 12849036 TI - Experiences of women with a diagnosis of breast cancer: a clinical pathway approach. AB - The study presented in this paper formed the first part of a large survey of breast cancer patients in one health authority in England, UK looking at individual needs expressed by women with a diagnosis of breast cancer. The paper provides an account of the experiences of 12 women with a diagnosis of breast cancer. The women represent a wide age range and different stages of illness. The transcribed accounts of the women were analysed by means of Qualitative Solutions and Research, Non-Numerical Unstructured Data Indexing Searching and Theorising (QSR*NUDIST). The study examined the individual experiences of women with a diagnosis of breast cancer and its aftermath as they passed through different stages related to it. The women's experiences are presented within the conceptual framework of the clinical pathway and their accounts represent their journey along the pathway. Various significant points in this journey are portrayed representing the women's reactions to diagnosis, treatment, femininity and body image, support, family and friends, information and after care. PMID- 12849037 TI - Nursing in Colorectal Cancer Initiative--the audit phase. Part 1. Development of the audit tool. AB - This paper describes the structure of an audit study together with the two-phase development of an audit tool and methodology. The purpose of the study was to develop a tool that defines and measures best practice in colorectal cancer nursing, and identifies areas for development. The European Oncology Nursing Society (EONS) and AstraZeneca collaborated to develop the Nursing in Colorectal Cancer Initiative (NICCI). The initiative was funded through educational grants from AstraZeneca and led by EONS. It is a project with two components, education and audit. The purpose of the project is to provide a sound educational basis for nurses in the elements of colorectal cancer care, together with the tools to evaluate and develop practice. The education phase culminated in an educational manual that provides a core set of materials concerning the fundamental aspects of colorectal cancer, to foster a common understanding amongst nurses nationally and internationally. The education manual provided the referenced rationale for the definition of a set of hallmarks and standards of clinical practice. The audit tool was developed from the hallmarks and standards and is focused on the care of patients receiving cytotoxic chemotherapy for colorectal cancer. The audit methodology is based on the structure, process outcome triad, together with the TELER system of audit. PMID- 12849038 TI - Pain experience following radical treatment for head and neck cancer. AB - During an investigation into the quality of life of people in the year following radical treatment for head and neck cancer, it became apparent that pain was a significant problem. Therefore, the current study was conducted to gain an understanding of the incidence and nature of pain in people who had received radical treatment for head and neck cancer and to explore their attitudes to pain and pain relief. A single cohort study was undertaken using a structured questionnaire designed for people with head and neck cancer (the EORTC QLQ-C30 and the EORTC QLQ H+N35). Fifty questionnaires were received from people who had received radical treatment between 6 and 12 months previously and who were disease-free. Twenty nine of the respondents also consented to a follow-up semi structured interview. The interviews generated qualitative data about personal experiences of pain and pain management in head and neck cancer. Patient recollections from interviews identified that only nine out of 29 (31%) interviewees had pain at diagnosis. However, 74% (37) of respondents to the questionnaire reported some degree of pain at follow-up. Patients having both surgery and radiotherapy were significantly more likely to have troublesome pain than patients who received radiotherapy only (Fisher's exact test=0.039). The qualitative data identified a wide variety of pain sites, a number of which were not covered by the EORTC tool. A significant number of patients were rejecting regular pharmacological management and using a variety of other pain-relieving measures. We conclude that through an understanding of pain experience and attitudes to pain management, nurses can facilitate the care and support of patients in pain following treatment for head and neck cancer. PMID- 12849039 TI - Nursing research and nursing practice: an exploration of two different cultures. AB - Despite increasing efforts by both the government and the professions to institute evidence-based practice, there still remains a significant research practice gap in UK nursing. This paper explores the two worlds of practice and research and the factors that may affect them. It suggests that academic researchers and practitioners often have different perspectives and are working under different imperatives. These different cultures need to be recognized and made more explicit. Consideration will be given as to what sort of evidence is required by practitioners to nurse effectively, how we should identify this evidence and the role which research plays. The disharmony between the worlds of research and practice extends the discussion to consider the ways in which research is produced and disseminated. The question is raised as to why researchers fail to identify the contingencies, adaptations and social biases that affect their work. Strategies that might facilitate a franker discussion of research and the way in which it is produced will be illustrated by examples of research work undertaken by the author. PMID- 12849040 TI - Children's cancer nurses sharing knowledge: the challenge of networking internationally. PMID- 12849041 TI - A study of factors influencing patients' decisions to undergo bone marrow transplantation from a sibling or matched related donor. AB - This study provides a theoretical analysis of factors influencing patients' decisions to undergo bone marrow transplantation. Data were gathered using single, unstructured interviews with seven bone marrow transplant candidates, and analysed in accordance with grounded theory methodology. Six theoretical categories were generated from the data. These were: the logical mind, gut instinct, personal aims, hope, preparing for transplant and trust in professionals. The findings are discussed in relation to theoretical literature and other research. The results of the study suggest that providing clinical or statistical information about transplantation may not, in itself, constitute adequate decision support. The adoption of a predominantly one-way process, whereby health-care professionals inform and educate patients, may fail to elicit broader subjective and contextual issues that influence patients' decisions. Patients may not fully consider treatment risks, or may interpret treatment information in idiosyncratic ways. Professionals may not foster adequate appreciation of risks, or may inadvertently frame information in ways that render some treatment options more acceptable than others. Recommendations focus upon: increasing professionals' awareness of the complex issues that may underpin patients' decisions, broader assessment of patients' understanding of decision situations, and tailoring information to meet the needs of individual patients. PMID- 12849044 TI - Evidence-based management of constipation in the oncology patient. AB - Constipation is suffered by over 50% of cancer patients and is regarded as one of the most distressing symptoms causing both physical and emotional distress. A need to treat constipation is often due to a failure to prevent it. There is little literature in this area and research based on clinical trials as to best treatment is virtually non-existent. It is suggested that many health-care professionals dismiss constipation as a relatively trivial problem, resulting in a lack of attention to the subject. There is a lack of consensus on the definition of constipation and confusion regarding effective methods for prevention and treatment. It could be argued that health-care professionals are more intent on monitoring the direct effects rather than the secondary effects of treatment. Since constipation is largely preventable, there is a need to highlight the importance of prevention in addition to establishing effective treatment guidelines. Oncology nurses are in an ideal position to identify cancer patients in a high-risk category and utilize preventive strategies. The study described has led to the development of evidence-based drug guidelines to be used in the prevention of constipation and acute/chronic constipation to ensure that patients receive the best treatment possible. PMID- 12849045 TI - The EONS core curriculum revision project. AB - This paper provides an overview of the EONS (European Oncology Nursing Study) core curriculum revision project. The aim of this project was to revise the EONS core curriculum for a Post-Basic Course in Cancer Nursing, first published in 1989, so that it would better reflect patient and family need and contemporary cancer nursing practice. A further aim of the project was to address the educational needs of both adult and paediatric oncology nurses. To achieve these aims a core curriculum revision meeting was organized and attended by representatives from most of the EU Member States. A rough draft of the core curriculum was developed, circulated and subjected to further scrutiny and modification. Following approval of the final draft, the core curriculum was translated from English into the 10 other official EU languages and made available in both paper and electronic form. The updated core curriculum was launched during the ECCO-10 conference and has now been disseminated widely amongst European cancer nurses and the wider nursing and medical community. PMID- 12849046 TI - Quality of life in cancer care: patients' experiences and nurses' contribution. AB - People with cancer experience great changes in their lives. Disease, treatment and prognosis affect their quality of life. The aim of this literature review was to ascertain how the illness affects patients' quality of life and to what extent nurses can influence or improve this experience. Twenty-two research articles were reviewed, the majority of which were qualitative in nature, studies that allow patients to portray their experiences in their own words. The concept of quality of life was defined as consisting of well-being in four core domains, psychological, social, physical and spiritual. It was found that patients' quality of life is affected in all four core domains and that these domains are overlapping and interdependent. The qualitative data also highlighted that each patient's experience is unique to that patient. Nurses have to be aware of this and remain sensitive to the individual patient's needs. Few of the articles reviewed examined nursing care from the patient's point of view. Tentative conclusions were that nurses can improve patients' quality of life by ensuring they are competent in daily practice, by giving patients a feeling of security and by treating them with respect as individuals and human beings. PMID- 12849047 TI - Men's experiences of their partner's primary and recurrent breast cancer. AB - The reaction and support from the partner of the woman with breast cancer is viewed by health professionals as an important factor in her adjustment to disease. However, there is little theory and research that focuses on the individual concerns and experiences of the man living with a woman who has breast cancer. Literature concerning the experience of breast cancer recurrence for both the partner and the patient is particularly sparse. The aim of this literature review is to explore the experiences of male partners of women with breast cancer. The role of the male partner supporting the woman with breast cancer, and the potential concerns he may have, will be explored. It will also focus on the partners' experience of living with a woman with primary breast cancer. Finally, the partners' experience of living with a woman with a recurrence of metastatic breast cancer will be considered. Relevant literature is reviewed and critiqued to place current knowledge in context, including identifying areas for further research. PMID- 12849048 TI - Oncology out-patients nursing: a challenge within the changing face of cancer care. AB - Eighty to 90% of cancer care is delivered in the out-patient setting, and thus the out-patient nurse's role is pivotal to patient care. A report published in 1990 (NHS Management Executive) suggested that there had been no change in the traditional 'handmaiden' role of the out-patient nurse over the previous 20 years. This study was designed using the conceptual framework of the domains of ambulatory nursing practice developed by Verran (1981). Two self-report questionnaires were developed in order to identify what nurses were actually doing in out-patients and the activities which both nurses and doctors felt should be undertaken by nurses. Questionnaires were distributed to all out patient nurses, clinical nurse specialists and doctors working within an oncology out-patients department in a London teaching hospital. Results showed that oncology-trained nurses were more involved in areas where specialist skills and knowledge are important for patient care. However, there were still non-nursing aspects of care that could be relinquished. Both nurses and doctors saw the role of the oncology-trained nurse as important in undertaking professional nursing activities within the out-patient department. This study suggests that specialist oncology out-patient nurses are beginning to break away from the traditional 'handmaiden' nurse's role utilizing their knowledge and skills within these departments. However, they still have to develop the role further in order to meet the needs of patients and their families within the out-patient setting. PMID- 12849049 TI - Fatigue patterns in Chinese patients receiving chemotherapy. AB - A descriptive study was conducted to explore the patterns, risk factors and experience of fatigue in Chinese cancer patients receiving chemotherapy. Forty two adult patients from an out-patient clinic of a university hospital in Hong Kong participated in the study. They were asked to complete a diary over a period of 2 weeks from the beginning of their chemotherapy cycle. The diary incorporated a 10cm horizontal visual analogue scale (VAS) for measuring fatigue intensity and a marked space next to the VAS for recording body temperature. Further, the Chinese version of the fatigue subscale of the Profile of Mood States was completed before the chemotherapy session, at the end of the first week post treatment and at the end of the second week. Sociodemographic, physiological and disease/treatment-related information was obtained from the medical records. A semi-structured, open-ended interview was conducted with 18 of these patients. Most Chinese cancer patients (73.8%-90.5%) receiving chemotherapy in this study experienced fatigue. The peak level of fatigue was reported at day 3 post treatment and it was more evident in the evenings. Female patients, patients with stage I cancer and those with nasopharyngeal carcinoma reported higher levels of fatigue (P<0.05). No relationship was found between body weight and fatigue, and some association existed between febrile illness and fatigue (P<0.001). Qualitative data affirmed the pattern of fatigue intensity and various descriptions about the experience and coping techniques of fatigue were found. Individualized and repeated assessment of patients with fatigue, as well as correcting the treatable factors that contribute to fatigue are suggested. PMID- 12849052 TI - The "Autoimmune diseases" 40th anniversary. PMID- 12849053 TI - Pathogenic effects of anti-Fc gamma receptor IIIb (CD16) on polymorphonuclear neutrophils in non-organ-specific autoimmune diseases. AB - The receptors FcgammaRIIIb and FcgammaRIIa for the Fc portion of IgG are naturally expressed in polymorphonuclear neutrophils (PMN). Autoantibodies (Ab) against FcgammaRIIIb exist in patients with non-organ-specific disease. These may be categorized, based on the results of an indirect immunofluorescence (IIF) test for the detection of anti-membrane-bound FcgammaRIIIb autoAbs, and an enzyme linked immunosorbent assay for that of soluble FcgammaRIIIb-recognizing autoAbs. The IIF+ autoAbs are not cytotoxic, and prolong the survival of the cells. The autoAb-triggered anti-apoptotic signal may be transduced through FcgammaRIIa and/or CD11b, the beta-chain of the neighboring complement receptor type 3. However, FcgammaRIIIb appears to be as competent as FcgammaRIIa, because the results obtained using the respective monoclonal Abs are additive. Soluble FcgammaRIIIb binds to CD11b and produces similar effects, suggesting that autoAb stimulated FcgammaRIIIb can work in concert with CD11b. Anti-FcgammaRIIIb conditioned supernatant of PMNs induces the transcription of messenger RNA for granulocyte colony-stimulating factor (CSF) and granulocyte-macrophage CSF, followed by the release of these anti-apoptotic factors. The delay in apoptosis is accompanied by a down-regulated expression of Bax. Thus, apoptosis of aged PMNs can be modulated by signaling through FcgammaRIIIb, which may occur in patients with PMN-binding anti-FcgammaRIIIb autoAbs. PMID- 12849054 TI - Immunomodulation for treatment and prevention of atherosclerosis. AB - In recent years, it has become clear that the immune system has a major role in atherosclerosis development and progression. More specifically, atherosclerosis fulfills all four criteria to define a condition as being an autoimmune in nature. This association also suggests that modification of the immune response in atherosclerosis could affect this process. In this review, we summarize different aspects of immunomodulation in atherosclerosis. These include immunosuppression, active immunization, induction of tolerance, administration of intravenous immunoglobulin, gene therapy, cytokine network manipulation and statins. PMID- 12849055 TI - Pathogenesis of autoimmune diseases associated with 8.1 ancestral haplotype: effect of multiple gene interactions. AB - Genetic studies have shown that individuals with certain HLA alleles have a higher risk of specific autoimmune disease than those without these alleles. Particularly, the association in all Caucasian populations of an impressive number of autoimmune diseases with genes from the HLA-B8,DR3 haplotype that is part of the ancestral haplotype (AH) 8.1 HLA-A1, Cw7, B8, TNFAB*a2b3, TNFN*S, C2*C, Bf*s, C4A*Q0, C4B*1, DRB1*0301, DRB3*0101, DQA1*0501, DQB1*0201 has been reported by different research groups. This haplotype, the more common one in northern Europe, is also associated in healthy subjects with a number of immune system dysfunctions. It has been proposed that a small number of genes within the 8.1 AH modify immune responsiveness and hence affect multiple immunopathological diseases. In this paper, the characteristic features of this haplotype that might give rise to these diverse conditions are reviewed, focusing on the role of multiple gene interactions in disease susceptibility of 8.1 AH. PMID- 12849056 TI - Antimitochondrial antibodies in primary biliary cirrhosis: the role of xenobiotics. AB - Primary biliary cirrhosis is an enigmatic autoimmune disease of women characterized by antimitochondrial antibodies and destruction of intrahepatic bile ducts. The etiology of PBC is unknown, but we present data herein that the disease may be induced by xenobiotic (i.e. chemicals) exposure. In particular, we postulate that halogenated compounds will bind to the autoantigen, break tolerance, and lead to an intense mucosal response. PMID- 12849057 TI - Hughes syndrome crosses boundaries. AB - Hughes (antiphospholipid) syndrome is a condition in continuous evolution. Since it was first described almost 20 years ago, significant advances in its diagnosis and management have been made. Recently, classification criteria for definite antiphospholipid syndrome have been proposed and validated. However, there is still controversy regarding issues such as the utility of new diagnostic tests (detecting antibodies to beta(2)-glycoprotein I, phospholipids other than cardiolipin or phospholipid mixtures), the role of antiphospholipid antibodies in neurological conditions such as demyelinating disease, cognitive impairment or migraine and the optimal management of thrombosis and obstetric manifestations. These are all briefly discussed in this review. PMID- 12849058 TI - Clinical relevance of antiprothrombin antibodies. AB - Antiprotrombin antibodies belong to the family of antiphospholipid antibodies (aPLs). The clinical relevance of antiprothrombin antibodies has not been established and it depends on the applied detection method. Antibodies against phosphatidylserine-prothrombin complex (aPS/PT) are closely associated with clinical features of antiphospholipid syndrome (APS) and lupus anticoagulant rather than antibodies against prothrombin alone. The determination of aPS/PT in routine clinical practice should be done in conjunction with other aPLs detection to improve the likelihood of recognising the APS, which would ultimately facilitate the management of the disease. PMID- 12849059 TI - Endothelium as a target for anti-phospholipid antibodies and for therapeutical intervention. AB - Endothelium activation seems to represent one of the pathogenic mechanisms that induce the trombophilic state of the anti-phospholipid syndrome. The rationale behind such a statement lies on the demonstration that: (a) the major antigen of the anti-phospholipid antibodies (beta 2 glycoprotein I) can be expressed on the endothelial cell membrane, (b) the endothelial beta 2 glycoprotein I offers suitable epitopes for circulating antibodies, (c) the binding of anti-beta 2 glycoprotein I antibodies is capable to induce the appearance of a pro-coagulant and pro-inflammatory phenotype. Both in vitro and in vivo experimental models support such a hypothesis. Although a classical vasculitic process cannot be found in the anti-phospholipid syndrome there is indirect evidence that endothelial activation/damage does occur also in vivo. The demonstration that hydroxymethylglutaryl Co-enzyme A reductase enzyme inhibitors (statins) can block endothelial cell activation induced by anti-beta 2 glycoprotein I antibodies as well as by pro-inflammatory cytokines offers new therapeutical approaches. PMID- 12849060 TI - New insights into the pathogenesis of antineutrophil cytoplasmic autoantibody associated vasculitis. AB - Antineutrophil cytoplasmic autoantibodies (ANCA) directed to proteinase 3 (PR3 ANCA) or myeloperoxidase (MPO-ANCA) are closely associated with the idiopathic systemic necrotizing vasculitides, in particular Wegener's granulomatosis, microscopic polyangiitis and its renal limited manifestation, and Churg Strauss Syndrome. Many in vitro studies show that those ANCA have phlogistic potential, particularly at the interface of neutrophils and endothelial cells. A limited number of studies in experimental animals support their pathogenetic role. However, ANCA alone are not sufficient, as based on clinical and experimental data, and other, probably exogenous factors, seem necessary for disease induction and (re)activation. Among those silica and particularly, the carriage of Staphylococcus aureus have been proposed. Besides, various genetic factors are involved in disease susceptibility. The ANCA-associated vasculitides are systemic autoimmune diseases in which the interplay of autoimmunity with environmental and genetic factors determines their clinical expression. PMID- 12849061 TI - Neutrophil-specific autoantibodies in chronic inflammatory bowel diseases. AB - This review intends to highlight important differences between neutrophil specific autoantibodies (NSA) typically found in chronic inflammatory bowel diseases (CIBD) and anti-neutrophil cytoplasm antibodies (ANCA) associated with primary systemic small vessel vasculitides (SSVV). Indirect immunofluorescence (IF) techniques alone cannot distinguish NSA from ANCA and special measures must be taken to separate these two autoantibody populations. Many autoantigens originating in all cell compartments may be targeted by NSA in CIBD, several of these being constituents of neutrophil nuclei. Apart from the use of NSA in the differential diagnosis between Crohn's disease (CD) and ulcerative colitis (UC), very limited clinical significance is ascribed to these antibodies in CIBD. Laboratory reports on NSA-positivity must be clearly distinguishable from reports on ANCA to help avoid clinical misinterpretation. PMID- 12849062 TI - Is peripartum cardiomyopathy an organ-specific autoimmune disease? AB - Peripartum cardiomyopathy (PPCM) is a rare and serious heart disease that exclusively afflicts women during childbearing years. Symptoms include rapid onset of cardiovascular insufficiency occurring during pregnancy, initiated anytime between the third trimester until 5 months post-partum in the absence of any other signs or history of heart disease. The rare incidence of PPCM and the absence of any relevant animal models have limited research and understanding of the pathogenic mechanisms involved. Several compelling sets of data support the view that PPCM is a form of autoimmune IDCM. However, PPCM differs from autoimmune IDCM in that (a) it is associated with unique sets of autoantibodies and autoantigens, (b) it has a relatively rapid onset, and (c) it exclusively affects pregnant women. Furthermore, the etiology of PPCM is dependent on the interaction of pregnancy associated factors, e.g. increased hemodynamic stress, vasoactive hormones and fetal microchimerism, that co-operate in the context of essential immune and genetic environments for disease progression. Our model of PPCM attempts to represent how multiple factors, e.g. pregnancy, genetics, immune dysregulation, and fetal microchimerism are held in a complex dynamic balance that can co-operate towards the maintenance of cardiovascular health or disease in the mother (Fig. 1). A more thorough study of the precise nature of the cardiac tissue autoantigens may lead to the identification of the mechanisms of breakdown of self-tolerance and perhaps also the putative etiologic agent(s). Further studies of the precise nature of the cardiac tissue autoantigens and the specific factors governing the balance between tolerance and autoimmunity in the periphery, e.g. expression of PD-L1 on cardiac tissues and the role of regulatory T cells, may help to elucidate the autoimmune mechanisms of PPCM. PMID- 12849063 TI - Recombinant autoantigens. AB - Autoimmune diseases frequently manifest themselves in fulminating immune reactions involving T and B cells that lead to the appearance of circulating autoantibodies. Such antibodies are considered to be of diagnostic relevance and may be pathognomonic for their associated autoimmune diseases. The advent of recombinant protein expression technology has made it possible to economically produce high quality autoantigens for use in immunoassays to detect autoantibodies in patients' sera. This short review is intended to give the reader a brief description of the various autoantigen production methodologies with their advantages and disadvantages, and the limitations of these techniques in the diagnostics of autoimmune diseases. PMID- 12849064 TI - Autoimmunization to epidermal growth factor, a component of the immunological homunculus. AB - Epidermal growth factor (EGF) is being tried as a vaccine in cancer immunotherapy with the aim of inducing neutralizing antibodies that might affect EGF-dependent tumors. Here we summarize our experience using the EGF self-molecule as an autoimmunigen. We report here that IgG anti-EGF antibodies are prevalent in healthy people and that augmentation of the response to EGF requires conjugation to an effective carrier and an adjuvant. Paradoxically, the response to EGF immunization could be enhanced by an 'immunosuppressive' treatment with cyclophosphamide, most probably by suppressing active control mechanisms. EGF is expressed in the thymus. Thus, EGF may be added to the immunological homunculus, the class of self-antigens to which there is both natural autoimmunity and natural regulation of the autoimmunity. The results using EGF as a vaccine can teach us about the homunculus and how to activate it. PMID- 12849065 TI - Iodine: an environmental trigger of thyroiditis. AB - Like most autoimmune diseases of humans, chronic lymphocytic (Hashimoto's) thyroiditis results from the combination of a genetic predisposition and an environmental trigger. A body of clinical and epidemiologic evidence points to excessive ingestion of iodine as an environmental agent. In genetically determined thyroiditis in animals, iodine enrichment has been shown to increase the incidence and severity of disease. Its mechanism of action is still uncertain. Using a new animal model of autoimmune thyroiditis, the NOD.H2(h4) mouse, we have been able to show that iodine enhances disease in a dose-dependent manner. Immunochemical studies suggest that iodine incorporation in the thyroglobulin may augment the antigenicity of this molecule by increasing the affinity of its determinants for the T-cell receptor or the MHC-presenting molecule either altering antigen processing or by affecting antigen presentation. PMID- 12849066 TI - Autoantibodies to factor VIII. AB - Anti-Factor VIII (FVIII) antibodies represent a unique model to study the relationship between natural autoreactivity (natural antibodies to FVIII of healthy individuals) and disease-associated autoimmunity ('spontaneous' FVIII inhibitors of patients with anti-FVIII autoimmune disease) to a single human protein antigen. Although natural and disease-associated anti-FVIII antibodies are not readily distinguished based on the comparison of their isotypic distribution and epitope mapping, available studies of cross-reacting idiotypes suggest that FVIII inhibitors in patient's plasma encompass two populations of anti-FVIII antibodies - some antibodies result from the clonal expansion of B lymphocytes that exist previous to the treatment with FVIII and secrete anti FVIII antibodies with properties similar to those of natural anti-FVIII antibodies present in healthy individuals, other inhibitors are produced by B cell clones that have undergone affinity-maturation and hypermutation of the V regions of the antibodies they produce. The implications for the treatment of autoimmune patients with anti-FVIII inhibitors are discussed. PMID- 12849067 TI - High-density lipoprotein-associated apolipoprotein A-I: the missing link between infection and chronic inflammation? AB - The etiology of chronic immuno-inflammatory diseases including rheumatoid arthritis (RA), multiple sclerosis (MS), systemic lupus erythematosus (SLE), and atherosclerosis is far from being elucidated. It is generally accepted that multiple factors are involved in the development of such pathologies, including factors of genetic susceptibility that interact in complex ways with diverse environmental factors, i.e. gender, nutrition, environment, etc. Furthermore, infection has often been pinpointed as playing a causal role. However, no distinctive pattern has yet emerged from the tremendous number of compiled results that would provide a generally acceptable hypothesis of the etiology of immuno-inflammatory diseases, and the possibility of a persistent antigenic stimulus arising from an infection cannot be confirmed or refuted. At the cellular level, chronic inflammation is characterized by the infiltration of immuno-inflammatory cells into the target tissue, which mostly precedes tissue damage. At the inflammatory site, monocytes and T lymphocytes are in close proximity. We have demonstrated that contact-mediated activation of monocytes by stimulated T lymphocytes is a major stimulus triggering the production of large amounts of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL 1beta) whose importance in chronic inflammation is well known. We recently established that high-density lipolipoprotein (HDL)-associated apolipoprotein (apo) A-I is a specific inhibitor of cytokine production in monocyte-macrophages upon contact with stimulated T cells. HDL-associated apo A-I is a negative acute phase protein, i.e. a protein whose level is lowered by more than 25% during the acute phase. This review aims at highlighting the fact that HDL-associated apo A I might play the role of a constitutive anti-inflammatory factor. The decrease of plasma levels of HDL-associated apo A-I upon acute inflammation may be a sign of the possible development of chronic inflammation, i.e. individuals presenting with risk factors might develop chronic inflammatory diseases after infection. We thus hypothesize that HDL-associated apo A-I might be the missing link between infection and chronic inflammation. PMID- 12849068 TI - The health status of community based elderly in the United Arab Emirates. AB - Little is known about the elderly in the United Arab Emirates (UAE), a country with both developing country features (high fertility rate, few elderly, strong traditional culture) and developed country characteristics (high-income economy, urbanized population, high growth rate of people aged 65+ years). In this cross sectional survey of 184 randomly chosen community based people aged 65+ years, the mean age was 71.8 +/- 6.3, 52% were female, 76% were married, 11% were literate, 89% lived in multi-generational households, 85% lived in households with servants and 15% had a personal servant. Health status was largely independent of age. Compared with the ambulatory aged USA population, the rate of functional independence in activities of daily living (ADL) (83%) was similar and chronic medical problems were less frequent, with the notable exception of diabetes (37% UAE, 10-12% USA). Almost all (95%) participants in this study rated their health as satisfactory or higher, compared with 82% of US ambulatory elderly. There appeared to be a significant under-diagnosis of psychological problems. In the presence of a high regard for traditional values, close family ties, universal practice of religion and high economic resources, the elderly in the UAE have a high level of health, which they maintain into their later years. There may be a need to substantially increase health care resources for aged care in the near future due to the high prevalence of diabetes, amount of hidden psychological morbidity and known demographic trends. Encouraging families to continue to provide home based long-term care may minimize the need for government intervention in this area. PMID- 12849069 TI - Aspects of quality of life in persons with pre-lingual deafness using sign language: subjective wellbeing, ill-health symptoms, depression and insomnia. AB - This study investigated health-related quality of life, expressed as subjective wellbeing and the prevalence of depressive symptoms and insomnia, among elderly pre-lingually deaf persons using sign language. Comparisons were made with elderly hearing people. Forty-five pre-lingually deaf persons, 65 years or older, took part (a response rate of 46%). Subjective wellbeing was assessed with the Gothenburg Quality of Life (GQL) instrument. Depressive symptoms were rated with the 15-item version of the geriatric depression scale (GDS), and insomnia was measured with Livingston's sleep scale. Ratings of subjective wellbeing among elderly pre-lingually deaf people were generally high. One third of the deaf persons demonstrated depressive symptoms and nearly two thirds suffered from insomnia. There was substantial correlation between insomnia, depressive symptoms and lower subjective wellbeing. The results strengthened the assumption that depressive symptoms and sleep disturbance are more frequent among elderly pre lingually deaf people using sign language than among hearing people. On the other hand, and contrary to our expectations, this did not imply significantly lower perceived subjective wellbeing compared with hearing elderly people. Results must be interpreted with caution due to limitations in the study. PMID- 12849070 TI - Effect of upper gastrointestinal endoscopy on cardiopulmonary changes in very old patients. AB - Although routine gastroscopy is regarded as a safe examination, it is sometimes associated, especially in elderly patients, with serious arrhythmias. We studied the influence of gastroscopy in very old patients on the occurrence and number of ECG changes and on oxygen saturation. Electrocardiographic (ECG) changes on a 24 h Holter recording and arterial oxygen saturation (SaO2) changes measured by pulse oximetry were observed in 37 hospitalized patients aged 80 years or more undergoing gastroscopy without premedication or conscious sedation and with supplementary oxygen (2 l/min). Gastroscopy did not induce significant arterial oxygen desaturation. ST changes were greatest or equal to that during gastroscopy in 16 (48%) patients. The number of VES increased during the 1-h period after gastroscopy, especially in those patients with an ST level change of over 1 mm h after gastroscopy (P=0.01) and in patients suffering from heart disease (P=0.007). In other arrhythmias no significant change was observed and no fatal complications occurred. Gastroscopy is a safe procedure also in very old patients. However, it induces increased number of VES after endoscopy in patients suffering from heart disease. In those patients a close follow-up of adverse symptoms is advisable, also for a short period after gastroscopy. PMID- 12849071 TI - Study of respiratory function in the elderly with different nutritional and cognitive status and functional ability assessed by plethysmographic and spirometric parameters. AB - From a sample of 265 elderly subjects (age 80.2 +/- 6.8 years) admitted to a geriatric care unit, free from cardiac and respiratory diseases, with normal chest X-ray and gas analysis, 53 subjects were selected and their respiratory functions (assessed by spirometric parameters and airway resistance 'Raw') were studied to correlate them with nutritional status, cognitive impairment, independence in everyday life activities and mood disorders, assessed, respectively, by the Mini Nutritional Assessment, rapid Mini Mental State test (MMSr), activities of daily living (ADL), instrumental activities of daily living (IADL) evaluation, Barthel Index and Cornell Depression Scale. The enrolled subjects were able to perform normally a forced expiration, although most of them committed errors in this test, according to the American Thoracic Society (ATS) criteria. Thus, about 32% started at lung volume0.12 s); in 62% of the subjects expiration time was too short and in 58% the terminal plateau was <2 s. Nevertheless, the spirometric parameters (and Raw) were considered normal. Forced vital capacity and peak expiratory flow (but not FEV1) were higher in the subjects without disability, while cognitive and nutritional status did not seem to have any influence on spirometric performance; MMSr score was related to compliance with ATS criteria for acceptability of the forced manoeuvre; mental and mood disorders, nutritional conditions and disability did not seem to have any influence on error rates. Our data show that our geriatric enrolled patients were able to perform an imperfect, often unfinished, but acceptable forced expiration; dynamic index values were related to disability, while the errors in starting the test were related to the mental conditions. PMID- 12849072 TI - Upper gastrointestinal endoscopy in patients aged 85 years or more. Results of a feasibility study in a district general hospital. AB - We performed a cross sectional analysis of the feasibility and yield of upper gastrointestinal endoscopy (UGE) in a cohort of patients aged 85 years or more. The study involved 218 patients who underwent diagnostic upper gastrointestinal endoscopy in a district general hospital between 1994 and 1998. Indication, use of sedation, endoscopic findings and treatment after endoscopy were evaluated. Indications for gastroscopy were suspicious of upper gastrointestinal bleeding (UGI) bleeding (41%), anemia (15%), and presence of dyspeptic- (31%), alarm- (9%) and/or reflux symptoms (3%). Serious UGI disease (cancer, peptic ulcer, reflux oesofagitis and/or erosive gastritis/duodenitis) was detected in 97 patients (44%). With respect to clinical presentation, serious UGI disease was present in 61% with bleeding, in 57% with reflux symptoms, in 42% with alarm symptoms, in 33% with anemia and in 28% with dyspepsia. Carcinoma was detected in eight patients (3.8%), all of them were treated with supportive care. In very old people gastroscopy is generally performed on sound indications reveals serious UGI disease in almost one out of two patients, markedly influences medical treatment, and reveals low malignancy rates (3.8%). In these patients, UGE is worthwhile and should not be omitted because of age considerations. PMID- 12849073 TI - The role of lipid profile in determining the risk of ischemic stroke in the elderly: a case-control study. AB - In this study, we investigated the association of lipids with ischemic stroke and its different subtypes in elderly patients. In particular, lipid parameters not extensively investigated so far in previous case-control studies specifically focused in the old population, such as lipoprotein Lp (a) and Apoproteins AI (ApoAI) and B (ApoB), have been taken into account. Seventy nine patients (mean age 83 +/- 7.4, range 67-99), consecutively admitted to a Geriatric Ward between January 1998 and June 2000 with acute stroke (first event) were studied. A complete clinical and laboratory assessment, including neurological evaluation, head CT scan, carotid ultrasonography and ECG, was employed to define the clinical and etiologic stroke subtype, according to standardized criteria. Fasting blood samples were collected within 48 h from admission, for determination of total cholesterol (TC), triglycerides (TG), High Density Lipoprotein-cholesterol (HDL-C), Lp(a), ApoAI and ApoB; Low Density Lipoprotein Cholesterol (LDL-C) was estimated by Friedwald formula. Eighty eight age and sex matched outpatients, referred to the hospital for non-inflammatory disorders of joints and musculoskeletal system, served as controls. Patients showed HDL-C and HDL-C/ApoAI ratio significantly lower than controls, with higher LDL-C/HDL-C ratio. Analysis on quartiles of lipoprotein concentrations showed also a significant increase in odds of stroke for LDL-C concentrations over 100 mg/dl, in absence of a linear relationship between LDL-C levels and risk. Multiple logistic regression, adjusting for non-lipid risk factors for stroke, confirmed the independent association of low HDL-C and HDL-C/ApoAI with all strokes, as well as with each subtype. In conclusion, these data suggest that lipids give some contribution to stroke risk even in the elderly, with a more prevalent role for HDL than LDL, and that lipid profile assessment must be taken into account in estimating the individual risk of stroke. PMID- 12849074 TI - A comparison of correlates of self-rated health and functional disability of older persons in the Far East: Japan and Korea. AB - Self-rated health and physical functioning are recognized as important indicators of health in older persons. Rarely, however, there have been studies done which examine cross-cultural differences in the health of older people using these measures, especially among non-Western countries. The objective of this study was to examine patterns of association of self-rated health and functional disability of Japanese and Korean elderly people living in the community, using nationwide surveys of persons aged 60 years or over. There were striking similarities in the general pattern of associations with covariates. In the multivariate analysis, age, work status, comorbidity, depressive symptoms, life satisfaction, hospitalization, and functional disability were strongly associated with self rated health in both populations. For functional disability, older age, female, low social contact, depressive symptoms, poor life satisfaction, and poor self rated health were found to be significantly associated. Some differences in the structure of associations with self-rated health, however, were noted. Women tended to assess their health more favorably than men in Korea, but in the Japanese elderly gender differences disappeared when other variables were taken into account. Health-related variables tended to be more closely associated with functional disability in the Japanese sample. An overall similarity, however, in the pattern of associations of these measures supports their utility in assessing and comparing the health of older populations in this region. PMID- 12849075 TI - Gender and rural-urban differences in reported health status by older people in Bangladesh. AB - The study aims to (i) describe regional variation and gender differences in health status of older people (60 years and older) in Bangladesh, indicated by self-reported health problems and functional ability; (ii) explore influence of socio-economic factors on health status of older people. In a cross-sectional study in rural and urban Bangladesh, 696 older persons were asked about their health problems and ability to manage activities of daily living (ADL). More than 95% of older people reported health problems. Approximately 80% of elderly women in both the regions reported having four or more health problems compared with 42% and 63% elderly men in the urban and rural regions, respectively. More women (urban: 55%; rural: 36%) than men (urban: 32%; rural: 22%) also reported difficulties with ADL. Irrespective of age, sex and area of residence, those reporting greater number of health problems were more likely to report difficulty with at least one ADL task. Reporting pattern of specific health problems varied between urban and rural regions. Socio-economic indicators were found to have little influence on reporting of health problems, particularly in the rural region. Observed regional difference may be related to the influence of social and environmental factors, and level of awareness concerning certain health conditions. PMID- 12849076 TI - Functional results after patellar fractures in elderly patients. AB - Elderly patients tend to fracture the patella because of a simple fall. In this group of patients restoration of primary functional level is crucial. Sixty-eight patients more than 65 years of age with patellar fractures, who were treated in our institution between January 1990 and December 1999 were evaluated. There were 43 females and 25 males. The ages ranged from 65 to 88 years (17 patients>80). We focus on the mechanism, which led to the fracture, the pre-morbid and post-morbid state of the patients and the various aspects of treatment in this group of patients. The majority (82%) had a simple fall. Half of the patients had background diseases. Forty-five patients (66%) had comminuted patellar fracture. In 58 patients (85%) the extensor mechanism was disrupted, which therefore, necessitated an operation (group I). The rest, ten patients, were treated conservatively (group II). Follow-up time ranged between 0.5 and 10.5 years (mean 4.5 years). Minor complications were noted in five patients. Sixty patients (88%) had intact extensor mechanism, while five patients in group I (9%) and three in group II (30%) had extension-lag of 10-30 degrees (P<0.05). None of the patients had non-union, cosmetic disturbances or developed osteoarthritis. Fifty-six patients (82%) (41 patients in group I and five in group II, P<0.05) achieved the same independence mobility status post-fracture. In elderly patients the cause of the fracture is a direct blow by simple fall to the patella causing comminuted fractures; however, with these patients excellent or good results were eventually achieved. Surgical treatment yielded better results than non-surgical. PMID- 12849077 TI - A positive role of phosphatidylinositol 3-kinase in aging phenotype expression in cultured human diploid fibroblasts. AB - In order to detect the role that phosphatidylinositol 3-kinase (PI3K) plays in the aging of human diploid fibroblasts, we analyzed cellular inositol phospholipids and expression of PI3Ks. In aged cells a decrease in phosphatidylinositol 3,4-bisphosphate (PI3,4P(2)) was notable, while phosphatidylinositol 3-phosphate (PI3P) and phosphatidylinositol 4,5-bisphosphate (PI4,5P(2)) decreased slightly. On the other hand, the messages of PI3K IIalpha, Vps34, and p110delta decreased and that of PI3K IIbeta increased during aging. These changes might relate to the aging phenomena, with the PI3K subspecies functioning differentially. Consistently, a PI3K inhibitor LY294002 greatly suppressed enlargement and flattening of cell body and nucleus as well as cell proliferation, both phenotypes being typical of aged cells. An oxidative stress, pulse exposure to hydrogen peroxide (H(2)O(2)), induced these senescent cell-like phenotypes, which LY294002 was also able to abolish. Upon examining three different cell systems (HL-60, N1E-115, and PC-12 cells) we found clear parallelism in a cellular event between the dependence on a PI3K activity and the sensitivity to H(2)O(2). On the analogy of these relationships, we could hypothesize that expression of an aging phenotype such as the morphogenesis is positively promoted by some PI3K subspecies, if such a phenotype as cell cycling is negatively affected by attenuation of another PI3K function in the course of cellular aging. PMID- 12849078 TI - Pyridoindole antioxidant stobadine protected bovine serum albumin against the hydroxyl radical mediated cross-linking in vitro. AB - On exposure to free radicals generated by the Fenton reaction system of Fe(2+)/EDTA/H(2)O(2)/ascorbate, bovine serum albumin (BSA), used as a model of water-soluble protein, was losing its water solubility depending on the concentration of the chelated iron. The precipitate was found irreversibly insoluble even in concentrated urea. In the soluble fraction, SDS-PAGE analysis proved the presence of dimers and trimers of BSA, accompanied by enhanced bityrosine fluorescence. The pyridoindole antioxidant stobadine inhibited the process of albumin insolubilization in a concentration-dependent manner, the protective effect being more efficient than that of 2-keto-4-methiolbutyric acid (KMBA). Stobadine was, however, less effective than trolox. The inhibitory effect of the antioxidants, expressed as IC(50), correlated well with the reciprocal values of corresponding second order rate constants for scavenging OHz.rad; radicals. The results indicated that the insolubilization of BSA induced by the Fenton system of Fe(2+)/EDTA/H(2)O(2)/ascorbate was caused by OHz.rad; radical mediated cross-linking of the albumin. The model system proved to be suitable for convenient testing of OHz.rad; radical scavenging ability of new antioxidants in a non-lipid environment. PMID- 12849079 TI - Regional cerebral blood flow and EEG in clinically diagnosed dementia with Lewy bodies and Alzheimer's disease. AB - This study was undertaken in order to compare regional cerebral blood flow (rCBF) and EEG findings of patients with clinically diagnosed dementia with Lewy bodies (clinDLB) and Alzheimer's disease (clinAD). Furthermore, within the clinDLB group to compare cases with and without neuropathologically verified Lewy bodies (LBs). When we studied 200 dementia cases in a prospective longitudinal dementia study, 48 had clinDLB and 45 clinAD in retrospective analyses. EEG information was analysed in 34 clinDLB and 28 clinAD patients and cerebral blood flow, measured with the Xe 133 inhalation method, in 26 clinDLB and 25 clinAD. There were no differences in EEG between the clinDLB and clinAD groups or between the cases with and without LBs. The rCBF patterns in the clinDLB and clinAD groups showed similar reductions in the temporoparietal areas. The rCBF in cases with LBs showed heterogeneous pathology. The imaging results in clinDLB and clinAD were strikingly similar. The EEG and rCBF could not differentiate between cases with or without LB. The study illustrates the lack of specific changes of EEG and rCBF in cases with LB pathology. PMID- 12849080 TI - Screaming, shrieking and muttering: the noise-makers amongst dementia patients. AB - We have described twelve dementia patients with noise making. We categorized noise making into (i) persistent screaming, (ii) perseverative vocalization, (iii) continuous chattering, muttering, singing or humming, and (iv) swearing, grunting and bizarre noise-making. The patients' ages ranged from 70 to 92 years with a mean of 78. There were four males and eight females. Five had Alzheimer's disease and the remaining seven vascular dementia. Five exhibited mild to moderate aggression while another five displayed severe aggression. All but one had motor restlessness and four had delusions or hallucinations. In ten patients the dementia was moderate to severe with five having total dissolution of speech. All but two were in the third stage of the illness and were totally dependent for basic self care. There was a wide network of anatomical structures involved in the twelve patients. The findings are related to what is known of the neuroanatomic location of the lesions and the role of frontal subcortical circuitry and neurotransmitter systems in relation to behavior. Noise-making could be explained by damage to the frontal lobe or interruption of the complex subcortical circuits and related brain chemistry. Treatment of patients with noise-making remains a challenge. PMID- 12849081 TI - Effects of a nursing intervention program on disabled patients and their caregivers. AB - This study evaluated a continuous home-visiting program designed for disabled patients. An intervention program was also designed for caregivers that included education and disabled care skills training, and emotional support over a 22-week period. The criteria for the disabled patients were as follows: (1) aged 18 years or older, (2) Barthel activity of daily living (ADL) score of <80 on the first visit, and (3) onset of disability prior to study was less than 2 years. A total of 126 participants were transferred from several hospitals to the community to be cared for by caregivers. Patients were divided into two groups: control group (n=56), and intervention group (n=70). In the control group, caregivers were not provided with any special training and patients were just evaluated at the beginning and end of the program. In the intervention group, caregivers were given intensive training in care for the disabled and completed eight home visits. The outcomes were measured before and after program using Barthel ADL index, life satisfaction scale (LSS) and caregiver's burnout scale (CBS). The results showed that improvements in ADL, LSS and CBS were significantly greater in the intervention group, compared to the control group. Multiple linear regression models showed that improvements in ADL were higher among patients with early-stage (onset of disability was less than 6 months before the start of the program) disability, patients whose caregivers had good health status, patients in the intervention group and patients who received more daily care hours. For LSS, there was a negative correlation with change in CBS and financial burden. In conclusion, the intervention program significantly improved disabled patients' ADL scores, as well as caregivers' LSS and CBS scores. It is necessary to coordinate medical and public health resources in the community to provide disabled patients and their caregivers with specific training and caregiving skills. PMID- 12849082 TI - Surgical management of spinal stenosis: a comparison of immediate and long term outcome in two geriatric patient populations. AB - This retrospective study was performed to assess and compare the immediate safety and long term outcome of surgical decompression of spinal stenosis when performed for geriatric patients aged 65-74 years (Group A) and those >75 years (Group B) of age. Some 283 patients were studied (179 and 104 patients in Groups A and B, respectively). The mean follow-up time was 41.5 months (range: 9-115) and 42.9 months (range: 9-106) for Groups A and B, respectively. Within both treatment groups, there was a significant (P<0.0001) subjective improvement in low back and radicular pain, as well as the ability to perform daily activities. When compared to preoperative levels, the oral scores for pain while performing daily activities were significantly (P<0.001) improved in both treatment groups. The overall postoperative complication rate was similar between the groups. Age is not a contraindication for decompressive lumbar spine surgery. However, since both Group A and B patients are likely to suffer minor perioperative complications, increased vigilance and careful monitoring are essential for this high risk population. PMID- 12849083 TI - High serum TSH levels are associated with depression in the elderly. AB - In order to investigate the association between elevated serum TSH levels and depression in the elderly, we conducted a population-based study of 451 over 60 year-old outpatients of a general University Hospital. Patients were divided into Group I (GI) (248 individuals) with high serum TSH levels, but otherwise no important condition or disease, and Group II (GII) (203 patients) with no previous diagnosis of thyroid or mood disease, referred to the hospital because of nonthyroidal severe diseases. All patients were clinically examined and classified according to DMS-IV for mood disturbance and had serum TSH, free T4 levels and antithyroid antibodies measured. High serum TSH levels (11.6+/-14.8 mU/l) were observed in 65/203 (32%) patients of GII. Among these patients, 42/65 (65%) had normal free T4 concentrations (1.23+/-0.98 ng/dl), no clinical manifestation of hypothyroidism and thus were considered to present subclinical hypothyroidism. Depression was observed in 24 cases from GI (9.7%) and 29 from GII (14.3%) and was frequent in the subclinical hypothyroid patients (49%). Our results suggest that mood disturbances are frequent in the elderly with elevated serum TSH levels, but they do not differ in the primary hypothyroid and the nonthyroidal sick patients. PMID- 12849084 TI - Screening for visual impairment in older people: validation of the Cardiff Acuity Test. AB - Poor visual acuity (VA) is a risk factor for falls, and a common impediment to rehabilitation, but conventional VA testing is difficult in dysphasic, deaf or confused patients. In the Cardiff acuity test (CAT) observation of the subject's eye movements (preferential looking) indicates if they can see a vanishing optotype on a card. The test is quick, and requires no speech or understanding on the part of the subject. We consider its usefulness in frail, elderly patients. Seventy-three patients aged 47-99 (mean 78) years, were tested in good lighting, wearing their usual spectacles. Cards ordered 'A' to 'K' with increasingly faint targets were sequentially presented at 1 m, until the subject's eye movements indicated the target to be invisible. We then performed conventional Snellen acuity chart testing. Twenty-three were retested by a second observer, and 24 subjects were retested by the same observer on another day. We used correlation coefficients to confirm inter-observer (r=0.95, P<0.01), and test-retest (r=0.97, P<0.01) reliability. Snellen chart measurements of VA were possible in all but six subjects. The results of CAT and Snellen acuity tests showed statistically significant correlation (r=0.35, P<0.01). The World Health Organisation (WHO) define significant impairment as a Snellen test acuity below 6/18, and using the 'G' card as a threshold were able to detect this degree of impairment with a sensitivity of 91.7%, and a specificity of 90.9%. The CAT is reliable, and highly acceptable to elderly patients, and may be useful as a screening tool in clinical practice, and for epidemiological purposes. PMID- 12849085 TI - Biological age--what is it and can it be measured? AB - Biological age is a concept used loosely and with little objectivity to describe a shortfall between a population cohort average life expectancy and the perceived life expectancy of an individual of the same age. Many biomarkers decline roughly linearly with age with a slope of <1% per annum. The use of a battery of 16 biomarkers is described as a method of calculating an individual biological age. They include: the concentration of prostacyclin in fibroblasts, cell membrane viscosity, the electroretinogram, baroreflex regulation of the heart rate, the concentration of lymphocytes, leucocyte density and velocity, grip strength, cells of the corneal endothelium and the buccal epithelium, neck muscle mobility, and vital capacity. Although not subjected to a prospective validation, the method might provide an objective approach to this widely used concept. PMID- 12849086 TI - Predictors of nursing home admission for older adults hospitalized with heart failure. AB - The objective of this retrospective chart review study was to determine the prevalence and predictors of nursing home admission of older hospitalized heart failure patients. Subjects were Medicare beneficiaries discharged with a principal diagnosis of heart failure in 1994 in the state of Alabama, United States. The outcome variable was admission to a nursing home after hospital discharge. Using multivariable logistic regression analyses we determined patient and care variables independently associated with admission to a nursing home. Patients (n = 985) had a mean (+/- S.D.) age of 79 (+/- 7.5) years, 61% were female and 18% African-American. Eighty-three (8%) patients were admitted to a nursing home. Over 80% of those admitted to a nursing home had prior nursing home residence. After adjustment for various demographic, clinical and care variables, age (adjusted odds ratio [OR] = 1.14; 95% confidence interval [95%CI] = 1.06 1.23), pre-admission residence in a nursing home (adjusted OR = 1422; 95%CI = 341 5923), and length of hospital stay (adjusted OR = 1.11; 95%CI = 1.02-1.20) were independently associated with admission to a nursing home. Among patients with no prior nursing home residency (n = 908), 15 (2%) patients were newly admitted to a nursing home upon discharge. In addition to age and length of stay, diabetes (adjusted OR = 6.46; 95%CI = 1.58-26.41) was independently associated with new admission to a nursing home. In conclusion, nursing home admission rate for this cohort of older hospitalized heart failure patients was low. Age, length of hospital stay, and diabetes were associated with new nursing home admissions. Further studies are needed to identify modifiable risk factors for nursing home admissions and to develop appropriate interventions. PMID- 12849087 TI - A longitudinal study on changes of movement performance and their relation to medical conditions in a female population followed from age 70 to 78. AB - We described longitudinal changes of movement performance in a population-based sample of women followed from age 70 to 78. We also studied the cross-sectional relationships between medical conditions and movement performance at baseline, and longitudinal relationships between baseline medical conditions and changes of movement performance. Two hundred and thirty-four women aged 70 years participated in the baseline study, and 88 women participated in a follow-up study 8 years later. Movement performance was measured by an optoelectronic test, the postural-locomotor-manual (PLM) test, which objectively and precisely measures the subject's mobility of lower and upper extremities. Information on medical conditions including selected diseases and symptoms were obtained by self report and/or by physical examination. Movement time (MT), an indicator of the overall movement performance of the PLM test, increased over 8 years. This change was mainly related to prolonged duration of the locomotor phase (walking forward), but not to the duration of the manual phase (goal-directed arm reaching). At baseline, poor PLM performance was related to hypertension, orthostatic hypotension, cerebrovascular diseases, chronic bronchitis, depression, arthritis, dizziness, chest pain, dyspnea, joint problems, leg pain, tiredness, number of diseases and number of symptoms at baseline. Increased MT during follow-up was associated with arthritis and dyspnea at baseline, and newly developed diseases during follow-up. Our study results indicated that 70-year-old women had a general slowing of their movement performance over 8 years. Age related decrements of movement performance were more striking in the lower extremities than in the upper extremities. Arthritis and dyspnea at baseline, and incident diseases during follow-up were related to this age-related decline of movement performance. PMID- 12849088 TI - Longitudinal changes in higher-level functional capacity of an older population living in a Japanese urban community. AB - This study examined the present state and longitudinal changes in higher-level functional capacity in a Japanese urban community. Persons aged 65-84 years living in a suburb of central Tokyo participated in a baseline survey held in 1991 (n = 814) and followed-up for 8 years. Outcome measures were disabilities in: instrumental self-maintenance (IADL), the intellectual activity scale (intellectual activity) and the social role scale (social role), as measured by subscales of the Tokyo Metropolitan Institute of Gerontology Index of Competence (TMIG-index of competence). At baseline among the three subscales, both older men and women had the highest prevalence of disability in social role, followed in turn by intellectual activity and IADL disability. The 8-year longitudinal survey on subjects who had no initial disability (229 men and 235 women) in all three subscales of TMIG-index of competence demonstrated that they were most likely to lose social role function with advancing age, followed in turn by intellectual activity and IADL. The Cox proportional hazard model analysis for all 814 participants revealed that baseline level of social role and intellectual activity significantly predicted the new onset of IADL disability during the 8 year follow-up period even after controlling for gender, age, and chronic medical conditions. In summary, disability in social role and intellectual activity do not only likely precede IADL disability, but also predict significantly the future onset of IADL disability in a Japanese urban community older population. PMID- 12849089 TI - Social and medical risk indicators for 8-year mortality in a Swedish urban elderly population. AB - The aim of this study was to identify and evaluate social and medical risk indicators for mortality in an urban elderly population. Altogether 217 subjects (144 women and 73 men, mean age 78 years, range 69-96 years of age) participated in an examination 1990/91. Eighty-eight persons (55 women and 33 men) had died, and 129 subjects (89 women and 40 men) were alive January 1, 1999. Several risk indicators were found and those with the highest statistical explanatory power to predict mortality were: tremor, inability for heavy housework, a pathological second heart sound, low triceps skinfold, low diastolic blood pressure and decreased appetite. A multivariate model (MVM) utililizing both social and medical risk indicators, and a clinical model (CM) based on the judgement of a registered nurse identified 49 and 34%, respectively, of those who died during the 8-year period (n = 88). A third risk group, the intervention group, comprising individuals selected by either the MVM or CM models, identified 56% of those who died. The latter procedure could be used to define risk groups for mortality in future intervention studies. The combination of social and medical risk indicators in MVM, and a CM might be used in studies with larger sample sizes in order to increase the knowledge in this field. PMID- 12849090 TI - Age-related changes in sensory and secretomotor nerve endings in the larynx of F344/N rat. AB - The aim of the present study was to define the age-related changes in sensory and secretomotor nerve endings in the larynx of F344/N rats. For this purpose, laryngeal tissue sections obtained from 12-, 24- and 35-month-old F344/N rats were compared with respect to the density, distribution and morphology of various types of sensory and secretomotor nerve endings immunoreactive for protein gene product 9.5 (PGP 9.5), calcitonin gene-related peptide (CGRP) and substance P (SP). Two distinct forms of PGP 9.5-immunoreactive motor end-plates were noted; the large sized motor end-plates localized in thyroarytenoid and cricoarytenoid muscles were degenerated in aged rats, while the small sized motor end-plates, localized predominantly in vocal muscles, did not show any age-related changes. CGRP- and SP-immunoreactive nerve fibres of the laryngeal glands did not show any age-related changes. Subepithelial laminar nerve endings immunoreactive to PGP 9.5 showed degeneration with ageing. Aggregates of terminal arborisations in the subepithelial region were smaller in aged animals. PGP 9.5-immunostained taste cells and well-developed subgemmal network were abundant in 12- and 24-month-old rats, but only a few were noted in aged rats. The total number of taste buds decreased significantly with ageing. CGRP- and SP-immunostained taste bud-nerve endings were noted in 12- and 24-month-old rats, but only rarely in 35-month-old rats. The laryngeal epithelium contained PGP 9.5-, CGRP- and SP-immunoreactive thin free nerve endings with many varicosities; their number and distribution were similar between 12- and 24-month-old rats, while only a few endings were observed in 35-month-old rats. Our results indicated that ageing is associated with the reduction of laryngeal sensory and secretomotor nerve endings. PMID- 12849091 TI - Long-term changes in mood of an aged population: repeated Zung-tests during a 10 year follow-up. AB - Associates and predictors of lowered mood were investigated in a 10-year prospective study of 411 random persons of three birth cohorts (aged 75, 80 and 85 years) in Helsinki, Finland. High Zung-score ( > 45 points = lowered mood) was found in 24% of subjects and clearly associated with age. Lowered mood was also associated with pessimistic attitudes towards life and impaired survival prognosis. The mean Zung-score fell drastically during the first follow-up (from 39.1 to 34.6 points, P < 0.001) and remained unchanged thereafter at 10-year examination of the survivors (33.9 points). Lowering mood (increase in Zung score) was best predicted by low baseline Zung-score (r = -0.673, P < 0.001), high baseline MMSE-score (r = -159. P < 0.05) and simultaneous changes in MMSE scores (r = 0.269, P < 0.01). The data show that lowered mood is associated with pessimistic attitudes towards life, cognitive impairment and impaired survival and that cognitive impairment exposes a patient to lowering mood. It is possible that a screening program improves the mood of aged population. PMID- 12849092 TI - Answering comments on the letter of J.O. Hornquist. An expression of concern on the paper 'Sleep and hypnotic use in relation to perceived somatic and mental health among the elderly' by Dr Ragnar Asplund (Arch. Gerontol. Geriatr. 31, 199 205 (2000)), which appeared in Arch. Gerontol. Geriatr. 35, 189-192 (2002). PMID- 12849093 TI - Suicides and the third age. AB - The phenomenon of suicide represents a complex problem, the specific aspects of which should be examined by a multifactorial analysis, particularly in the elderly subjects. Although the research on risk factors continues to grow, only a limited knowledge is available on the biological changes increasing the risk for suicide. Similarly, limited information is at our disposal about the contributing psychosocial processes extending beyond the demographic factors. Although the best explored population is the elderly using primary care services, no proven interventions are known for the time being, although some efforts to test certain approaches reaching these older adults are under way. Apparently even more, continued efforts are needed to change the attitudes toward the mental illnesses and their treatments in general, in order to reach the older adults who are still outside of the health care services. PMID- 12849094 TI - Alterations of the lymphocytic set-up in elderly patients with cancer. AB - It is assumed that the increased incidence of neoplastic pathologies with advancing age is correlated with the immunosenescence and with the altered immune surveillance. The present study was aimed at evaluating the role of the immunocompetent system and immunosenescence in carcinogenesis. A pool of 99 subjects (38 females, 61 males) has been analyzed in three groups as follows. Group A: 51 elderly subjects with cancer (16 females and 35 males, average age 73.7 +/- 7.5 years). Group B: 24 young subjects with cancer (12 females, 12 males, average age 49.5 +/- 10.3 years). Group C: 24 elderly subjects without any clinical evidence of cancer (10 females, 14 males, average age 74.6 +/- 6.3 years). Hemo-chromocytometric analysis and cytofluorimetric typifying have been performed in all subjects. A decrease of T (CD3+)-lymphocytes has been observed in group A, if compared to group B (P < 0.007), and to group C (P < 0.01), The T (CD4+)-lymphocytes were fewer in group A, than in group C (P < 0.004), and also the NK cells showed the same trend (P < 0.002). The numbers of leukocytes and monocytes increased in group A compared to group C (P < 0.01 and P < 0.004, respectively). Red cell numbers, hemoglobin and hematocrit values were lower in group A than in group B (P < 0.03, P < 0.03, P < 0.01, respectively), and also than in group C (P < 0.007, P < 0.001, P < 0.01, respectively), The results demonstrate that the alterations of the immunocompetent cells, particularly of the T-cell pool, may play an important role in the carcinogenesis of the elderly. PMID- 12849095 TI - Hospital readmission of persons with hip fracture following medical rehabilitation. AB - A significant percentage of older adults hospitalized and treated for hip fracture are readmitted to a hospital within six months. We analyzed information from a national database, the Uniform Data System for Medical Rehabilitation. Records for 8,236 patients (1994-98) who received inpatient medical rehabilitation following treatment for hip fracture were examined. Mean age was 76.51 years (S.D. = 12.48) with 71% female and 79% non-Hispanic White. The primary outcome measure was incidence of hospital readmission 0-180 days post discharge. The hospital readmission rate was 16.7%. A Cox regression model predicting rehospitalization included the following variables (p < 0.05): basic daily living skills, age, length of stay, ethnicity, and gender. There was a statistically significant difference in the percent of male versus female patients rehospitalized for Hispanic subjects but not for non-Hispanic white or African American subjects. The greatest variability occurred among male patients. A total of 18.1% of non-Hispanic White males and 16.8% of African American males were rehospitalized. In contrast, only 10.1% of Hispanic males were rehospitalized. Basic daily living skills, length of hospital stay, age, ethnicity and gender were variables associated with hospital readmission following medical rehabilitation in persons with hip fracture. These variables should be considered in developing intervention programs to reduce the risk of hospital readmission. PMID- 12849096 TI - Effect of age on the induction of 8-oxo-2'-deoxyguanosine-releasing enzyme in rat liver by gamma-ray irradiation. AB - Aged (27 months of age) and young (6 months of age) Fischer 344/DuCrj rats were exposed to gamma-ray irradiation, and their livers were compared for levels of oxidative DNA modifications and repair enzyme activities. The amounts of 8-oxo-2' deoxyguanosine (8-oxodG) in the nuclear DNA of the livers of both young and aged rats increased immediately after irradiation, by 1.7-fold in the livers of young rats and 2.7-fold in the livers of the aged rats. Also, the rate of 8-oxodG decay was slower in the livers of the aged rats than in young rat liver, and remained above the baseline level even 1 week after irradiation. The activities of 8-oxodG releasing enzymes peaked 2 and 6 h after irradiation in the livers of young and aged rats, respectively. The repair activity in the livers of the young rats was increased by sevenfold 2 h after irradiation, while the livers of the aged rats showed a twofold increase 6 h after irradiation. These results suggest that the ability to repair damaged DNA is lower in aged rats, and that the accumulation of oxidative DNA damage that takes place during aging may be related to this decline in repair activity. PMID- 12849097 TI - Ninety-seven-year-old people: general presentation, and some general and medical characteristics from a Swedish population study. AB - This study--the first in a series of reports--is a description of some general and medical characteristics of 97-year-olds from the representative longitudinal Gerontological and geriatric population studies in Goteborg, Sweden. The sample comprised 117 females and 15 males, a total of 132 97-year-olds. The probands were examined in their homes, with an interview based on a questionnaire. Blood hemoglobin, blood glucose and serum cobolamines were analyzed with standard methods, as well as a detailed hematological analysis which will be reported separately. An examination by a nurse and a physician was completed, comprising also dental status, visual ability and a simple hearing test. Anthropometric measurements and ECG were performed, and blood pressure was measured. Judged from cross-sectional data from our population studies, many symptoms and conditions seem to have a higher prevalence at age 97 than at age 70 and 85, respectively, especially regarding visual impairment, oral dryness, and hearing and ECG abnormalities. On the other hand the prevalence of edentulousness was surprisingly low. We find this investigation important for several reasons, namely its comprising a total community sample at this very high age in an urban area. Furthermore, the subjects are survivors of a cohort of elderly investigated several times by us since 1971/72 at age 70. In some aspects these survivors seem to be an elite of elderly people. PMID- 12849098 TI - Bacterial endocarditis of oral etiology in an elderly population. AB - The aim of this study was to analyze the prevalence and characteristics of bacterial endocarditis (BE) of oral origin in a group of elderly people. A retrospective study of 115 BE clinical records was performed, focusing on the demographic and predisposing features, as well as on the analytical and clinical variables. Twenty-two of the 115 cases were excluded as they were detected in intravenous drug users. Of the remaining 93 cases, 54.8% were diagnosed in patients older than 60 years of age (group A) and 45.2% in patients younger than 60 years (group B). There were 16 cases (17.2%) of oral origin; 4 BE cases mainly associated with tooth extractions were found in group A and 12 BE (most of them related with odontogenic abscesses) in group B. Within group A, 1 patient (25%) had not an underlying cardiac condition versus 5 cases (41.6%) in group B. Even though the prevalence of BE of oral origin in patients older than 60 is low, the high frequency of cardiopathies, poor oral health and high number of dental procedures shown by the old population makes them a risk group for BE of oral origin. PMID- 12849099 TI - Normal aging and AIDS. AB - Normal aging is accompanied by a wide range of changes in brain structure and function. In this work we seek to examine the hypothesis that normal aging results in neuropsychological findings similar to those produced by neuropsychological impairment related to AIDS. Specifically, we attempt to compare the level of neuropsychological performance in three groups: a normal elderly group (n = 46), a young group of AIDS patients (n = 36), and a young group of normal subjects (n = 22). The results indicated that, despite age differences between the normal elderly and AIDS patients, there was great similarity in the neuropsychological test performance of these two groups. In contrast, both groups demonstrated significantly different neuropsychological test performance than that evidenced by the younger normal controls. These results support the notion that neuropsychological changes related to normal aging are of a nature similar to those observed in AIDS and, therefore, affect functions subserved by frontal-subcortical brain regions. PMID- 12849100 TI - The unfavorable nature of preoperative delirium in elderly hip fractured patients. AB - The onset of delirium is frequent in elderly patients who sustain hip fractures. The purpose of this study was to characterize different patterns of preoperative and postoperative delirium, to study factors associated with preoperative delirium and to evaluate the possible different outcome of these patients. This retrospective study comprised 281 elderly patients with hip fractures undergoing surgical fixation. Data collection included age, sex, length of stay, type of fracture, cognitive status by mini mental state examination (MMSE), assessment of possible delirium by the confusion assessment method (CAM) and functional outcome assessed by functional independence measure (FIM). A database search was conducted to identify whether delirium onset occurred prior to or following surgery. About 31% of the total sample developed delirium. Delirious patients tended to be more disabled (P = 0.03) and cognitively impaired (P = 0.018), compared with non-delirious patients. Most delirious cases (53%) had their onset in the preoperative period. Patients with preoperative delirium were older (P = 0.03), had a lower prefracture mobility (P < 0.01), impaired cognition (P = 0.04) and showed an adverse functional outcome in terms of FIM score. Regression analysis showed that prefracture dementia, prefracture mobility and low MMSE scores were strongly associated with higher probability of having preoperative delirium, with no additional effect of other variables. It is concluded that preoperative delirium should be viewed as a separate entity with unfavorable nature and adverse outcome. Careful preventive measures and better treating strategies should be employed to avoid this clinical condition. PMID- 12849101 TI - Blood transfusion requirements in elderly hip fracture patients. AB - The purpose of this study was to investigate blood transfusion patterns in elderly hip fractured patients and to determine the clinical predictive criteria for blood utilization. This retrospective study involved data analysis of 302 patients, undergoing surgical repair of pertrochanteric and subcapital fractures. Patients of the transfused group had significantly lower admission hemoglobin (P < 0.0001) and significantly more pertrochanteric fractures. About 80% of patients of the lowermost quartile of admission hemoglobin were transfused, compared with < 20% of the uppermost group (P < 0.0001). Admission hemoglobin levels < 12 g% and pertrochanteric fractures were independently associated with transfusions (odds ratio (O.R.) 0.475, C.I. 0.377-0.598, P < 0.0001 and O.R. 1.885, C.I. 1.05 3.215, P = 0.02, respectively). The results support the adoption of a policy considering primarily these two factors. Other factors we studied had no predictive power. Cross-matching may be reserved only for patients presenting with hemoglobin < 12 g% and pertrochanteric fractures. This practice seems safe and should help in reducing the extent of unnecessary blood ordering. PMID- 12849102 TI - Post-hospitalization mortality in the elderly. AB - The level of disability and polypathology in hospitalized elderly is usually high. Multidimensional and functional assessment allows to identify risk factors for clinical and functional failure of patients. Many studies point out that identifying predictors of high-risk patients is a necessary step in accurate targeting. We evaluated 395 subjects (175 women, 202 men, mean age 77.9 year) during their hospitalization in our Geriatric ward. Baseline data included: demographics variables, medical diagnosis, functional evaluation, and laboratory values. After a 6-month follow up 80 (20.2%) subjects died. In our study, male gender, dependence at the Dependence Medical Index (DMI), low serum albumin (< 2.8 g/dl), impaired score at the Instrumental Activities of Daily Living scale (IADL), score lower than 13.7 at the acute physiology and chronic health evaluation (APACHE II) and neoplasm were independent predictors of 6-month post hospitalization mortality. The high mortality rate of our sample could be a marker of considerable frailty among elderly patients. Our study shows that a poor functional status is a more reliable prognostic factor than type and number of admitting diagnosis. Clinical evaluation, improved with information about functional status, is a feasible and practical way of detecting risk of short term post-hospitalization mortality of elderly subjects. PMID- 12849103 TI - Sleepiness and sleep in elderly subjects with hearing complaints. AB - The aim of this study was to evaluate the relation of hearing complaints to daytime sleepiness and sleep in a group of elderly men and women. A questionnaire survey was undertaken among 10,216 elderly subjects in northern Sweden. The mean (+/- S.D.) ages of the men and women were 73.0 +/- 6.0 and 72.6 +/- 6.7 years, respectively. Poor hearing was reported by 43.1% of the men and 22.8% of the women. The relative numbers with hearing complaints increased with age. Daytime sleepiness (DS) was reported by 32.9% of the men and 23.2% of the women. In a multiple logistic regression analysis significant independent correlates of daytime sleepiness in men were: hearing, good versus poor (odds ratio [OR] 1.5; confidence interval [CI] 1.2-1.8), health, good versus poor (3.3; 2.6-4.4), sleep, good versus poor (2.1; 1.6-2.8) and age, > or =80 years versus < 70 years (1.5; 1.1-2.1). The corresponding correlates in women were: hearing (1.4; 1.1 1.8), health (4.3; 3.4-5.4), sleep (2.2; 1.6-2.8) and age (1.6; 1.3-2.2). Thus, poor hearing was associated with increased daytime sleepiness independently of health, sleep status and age. Also subjects with hearing complaints more often slept poorly, woke up more often and had more difficulty in falling asleep after waking. PMID- 12849104 TI - Banking on regaining public confidence. PMID- 12849106 TI - 20000 brains removed and retained without relatives' consent. PMID- 12849107 TI - Cerebellar damage limits some, but not all, motor-control functions. PMID- 12849108 TI - D3 receptors and levodopa-induced dyskinesia. PMID- 12849109 TI - Abciximab extends treatment window for stroke. PMID- 12849112 TI - Spanish centre for rehabilitation of multiple sclerosis to be built. PMID- 12849115 TI - Neuroprotection for Parkinson's disease: a call for clinically driven experimental design. PMID- 12849116 TI - WHO will be "vigourously engaged" in neurological disorders. PMID- 12849117 TI - Supratentorial grade II astrocytoma: biological features and clinical course. AB - Because of its unpredictable clinical course, treatment strategies for low-grade (grade II) astrocytoma vary from "wait and see" to gross tumour resection followed by immediate radiotherapy. Clinical studies on grade II astrocytoma show that 5-year-survival ranges from 27% to 85% of patients with very few consistent prognostic variables besides the patient's age and the presence of neurological deficit. There is no universally recognised choice of therapy for patients with astrocytoma grade II, partly because of the shortcomings of histological classification systems. Routine microscopy tends to underestimate malignancy grading of astrocytomas and in most cases cannot distinguish between indolent and progressive subtypes. Recent studies suggest that proliferation and genetic markers can be used to identify subgroups of astrocytoma grade II with a rapid progressive clinical course. Therefore these markers should be included in ongoing and future clinical studies of patients with astrocytoma grade II. PMID- 12849118 TI - Interactions between antiepileptic and chemotherapeutic drugs. AB - Cancer and epilepsy commonly co-occur, and concomitant administration of antiepileptic (AEDS) and chemotherapeutic drugs (CTDs) is necessary in many cases. Many drugs are metabolised by the hepatic cytochrome P450 (CYP) isoenzyme system, and coadministration of AEDs and CTDs can lead to clinically relevant interactions by induction or inhibition of enzymes in shared metabolic pathways. These interactions can cause insufficient tumour and seizure control or lead to unforeseen toxicity. Enzyme-inducing AEDs reduce the effects of taxanes, vinca alkaloids, methotrexate, teniposide, and camptothecin analogues. Inhibition of the metabolism of nitrosoureas or etoposide by valproic acid can lead to CTD toxicity. Poor seizure control may result from the combinations of phenytoin with cisplatin or corticosteroids, and valproic acid with methotrexate. Increased toxicity of AEDs can occur when phenytoin is combined with 5-fluorouracil. Use of enzyme-inducing AEDs should be avoided in patients with cancer, particularly in association with chemotherapy. Generally, valproic acid-although not free from interactions-would be the agent of first choice. Some of the newer AEDs not metabolised by the P450 system may prove to be good alternatives. PMID- 12849119 TI - Therapeutic hypothermia for acute stroke. AB - Experimental evidence and clinical experience show that hypothermia protects the brain from damage during ischaemia. There is a growing hope that the prevention of fever in stroke will improve outcome and that hypothermia may be a therapeutic option for the treatment of stroke. Body temperature is directly related to stroke severity and outcome, and fever after stroke is associated with substantial increases in morbidity and mortality. Normalisation of temperature in acute stroke by antipyretics is generally recommended, although there is no direct evidence to support this treatment. Despite its obvious therapeutic potential, hypothermia as a form of neuroprotection for stroke has been investigated in only a few very small studies. Therapeutic hypothermia is feasible in acute stroke but owing to serious side-effects--such as hypotension, cardiac arrhythmia, and pneumonia--it is still thought of as experimental, and evidence of efficacy from clinical trials is needed. PMID- 12849120 TI - The production and use of cells as therapeutic agents in neurodegenerative diseases. AB - Although progressive neurodegenerative diseases have very different and highly specific causes, the dysfunction or loss of a vulnerable group of neurons is common to all these disorders and may allow the development of similar therapeutic approaches to the treatment of diseases such as amyotrophic lateral sclerosis, Parkinson's disease, and Huntington's disease. When a disease is diagnosed, the first step is to instigate protective measures to prevent further degeneration. However, most patients are symptom-free until almost all of the vulnerable cells have become dysfunctional or have died. There are known molecular mechanisms and processes in stem cells and progenitor cells that may be of use in the future design and selection of cell-based replacement therapies for neurological diseases. This review provides examples of conceptual and clinical problems that have been encountered in the development of cell-based treatments, and specific criteria for the effective use of cells in the future treatment of neurodegenerative diseases. PMID- 12849121 TI - Homocysteine and Alzheimer's disease. AB - BACKGROUND: A high circulating concentration of the amino acid homocysteine is an independent risk factor for stroke. Alzheimer's disease (AD) commonly co-occurs with stroke. Epidemiological studies found associations between hyperhomocysteinaemia and both histologically confirmed AD and disease progression and revealed that dementia in AD was associated with evidence of brain infarcts on autopsy. Thus, hyperhomocysteinaemia and AD could be linked by stroke or microvascular disease. However, given known relations between B-group vitamin deficiency and both hyperhomocysteinaemia and neurological dysfunction, direct causal mechanisms are also plausible. RECENT DEVELOPMENTS: A recent prospective study (S Seshadri and colleagues N Engl J Med; 2002 346: 476-83) showed hyperhomocysteinaemia to be a strong, independent risk factor for dementia and AD. The researchers found a graded increase in risk of both outcomes with rising plasma concentration of homocysteine after multivariate control for putative risk factors for AD. In conjunction with demonstration of a fall in homocysteine concentrations in response to increasing B-group-vitamin status, these findings give hope that mental decline, or AD itself, could be prevented by dietary modification or food fortification. WHERE NEXT? 25% of dementia cases are attributed to stroke. The possibility that some of the other 75% might be prevented by the lowering of homocysteine concentrations greatly increases the hope of maintaining self-sufficiency into old age. If homocysteine lowering can reduce the incidence of dementia or AD, decreased incidence of these disorders may be seen in Canada and the USA, where government-mandated folate-fortification programmes are in effect. Future research should focus on early detection of AD and on the possibility that the disease itself, or its primary symptom, could be prevented by folate supplementation. PMID- 12849122 TI - Convulsive ergotism: epidemics of the serotonin syndrome? AB - Between 1085 and 1927, epidemics of "convulsive ergotism" were widespread east of the Rhine in Europe due to consumption of grain contaminated with ergot, which is produced by the fungus Claviceps purpurea. West of the Rhine, consumption of ergot-contaminated food caused epidemics of gangrenous ergotism. The clinical features of convulsive ergotism--muscle twitching and spasms, changes in mental state, hallucinations, sweating, and fever lasting for several weeks--suggest serotonergic overstimulation of the CNS (ie, the serotonin syndrome). The ergot alkaloids are serotonin agonists. Dihydroergotamine binds to serotonin receptors in the dorsal horn of the spinal cord, which is the site of neuropathological changes in convulsive ergotism. Dihydroergotamine given to human beings can cause the serotonin syndrome. Ergots produced by different strains of Claviceps purpurea, and those growing in different soils, may have different ergot alkaloid compositions. An alkaloid, present in high concentrations in ergots from east of the Rhine, may have caused convulsive ergotism at a circulating concentration insufficient to produce peripheral ischaemia. The serotonin syndrome may, therefore, have been a public-health problem long before it was recognised as a complication of modern psychopharmacology. PMID- 12849123 TI - Paraneoplastic polymyositis in recurring adrenal gland carcinoma. PMID- 12849124 TI - Septo-optic dysplasia plus. PMID- 12849125 TI - Neural transplantation for the treatment of Parkinson's disease. PMID- 12849132 TI - Quantum mind. PMID- 12849133 TI - Clinical research on trial. PMID- 12849135 TI - Neurogenetics study was unethical, says NNI report. PMID- 12849136 TI - Neurophysiologist reprimanded by UK General Medical Council. PMID- 12849137 TI - US national plan to reduce stroke burden unveiled. PMID- 12849140 TI - Up Kv creek with a paddle. PMID- 12849144 TI - Drug resistance in epilepsy: not futile, but complex? PMID- 12849145 TI - Erythropoiesis and neuroprotection: two sides of the same coin? PMID- 12849146 TI - Alcohol consumption and the risk of dementia. PMID- 12849147 TI - Highs and lows of blood pressure:a cause of Alzheimer's disease? PMID- 12849148 TI - Not-so-silent infarcts. PMID- 12849150 TI - The use of quantitative magnetic-resonance-based techniques to monitor the evolution of multiple sclerosis. AB - Conventional MRI can improve accuracy in the diagnosis of multiple sclerosis (MS) and monitor the efficacy of experimental treatments. However, conventional MRI provides only gross estimates of the extent and nature of tissue damage associated with this disease. Other quantitative magnetic-resonance-based techniques have the potential to overcome the limitations of conventional MRI and, as a consequence, to improve our understanding of the natural history of MS. Magnetisation-transfer, diffusion-weighted, and functional MRI--as well as proton magnetic-resonance spectroscopy--are helping us to elucidate the mechanisms that underlie injury, repair, and functional adaptation in patients with MS. These techniques are substantially changing our understanding of how MS causes irreversible disability and should be used more extensively in clinical trials and in studies of disease progression. PMID- 12849151 TI - Clinically important drug interactions in epilepsy: general features and interactions between antiepileptic drugs. AB - There are two types of interactions between drugs, pharmacokinetic and pharmacodynamic. For antiepileptic drugs (AEDs), pharmacokinetic interactions are the most notable type, but pharmacodynamic interactions involving reciprocal potentiation of pharmacological effects at the site of action are also important. By far the most important pharmacokinetic interactions are those involving cytochrome P450 isoenzymes in hepatic metabolism. Among old generation AEDs, carbamazepine, phenytoin, phenobarbital, and primidone induce the activity of several enzymes involved in drug metabolism, leading to decreased plasma concentration and reduced pharmacological effect of drugs, which are substrates of the same enzymes (eg, tiagabine, valproic acid, lamotrigine, and topiramate). In contrast, the new AEDs gabapentin, lamotrigine, levetiracetam, tiagabine, topiramate, vigabatrin, and zonisamide do not induce the metabolism of other AEDs. Interactions involving enzyme inhibition include the increase in plasma concentrations of lamotrigine and phenobarbital caused by valproic acid. Among AEDs, the least potential interaction is associated with gabapentin and levetiracetam. PMID- 12849152 TI - Medical decompressive therapy for primary and metastatic intracranial tumours. AB - Medical decompressive therapy (MDT) with corticosteroids and mannitol is often used in patients with primary or metastatic brain tumours. This review highlights the lack of sound evidence regarding the indications and dosage schedule of steroids, prolonged use of which may cause debilitating complications. The available evidence supports the short-term use of MDT for raised intracranial pressure or progressive neurological deficits, but in the absence of these symptoms, MDT is not recommended for stable focal deficits, abnormal higher mental functions, seizures, or as prophylaxis during cranial irradiation. A practical stepladder guideline (based on symptom severity) is proposed with a starting daily dexamethasone dose of 6 mg for non-severe headache and or vomiting; 12 mg for progressive focal neurological deficit with or without non severe headache or vomiting; and 24 mg dexamethasone with mannitol for severe headache, vomiting, or altered consciousness. Depending on the clinical response, dose can be increased to the next step(s) or tapered every 48 h (more slowly in patients who are dependent on steroids). A scheme for the assessment of efficacy and toxicity prevention is also proposed. The proposed guidelines may be used as a template for further clinical research. PMID- 12849153 TI - Adenosine receptors and Huntington's disease: implications for pathogenesis and therapeutics. AB - Huntington's disease (HD) is a devastating hereditary neurodegenerative disorder, the progression of which cannot be prevented by any neuroprotective approach, despite major advances in the understanding of its pathogenesis. The study of several animal models of the disease has led to the discovery of both loss-of normal and gain-of-toxic functions of the mutated huntingtin protein and the elucidation of the mechanisms that underlie the formation of huntingtin aggregates and nuclear inclusions. Moreover, these models also provide good evidence of a role for excitotoxicity and mitochondrial metabolic impairments in striatal neuronal death. Adenosine has neuroprotective potential in both acute and chronic neurological disorders such as stroke or Parkinson's disease. Here we review experimental data on the role of A1 and A2A adenosine receptors in HD that warrant further investigation of the beneficial effects of A1 agonists and A2A antagonists in animal models of HD. Future pharmacological analysis of adenosine receptors could justify the use of A1 agonists and A2A antagonists for the treatment of HDin clinical trials. PMID- 12849154 TI - Consent to thrombolysis in acute ischaemic stroke: from trial to practice. AB - Thrombolysis with intravenous alteplase for acute ischaemic stroke, which is already in clinical use in the USA and Canada, has recently been approved in the European Union. Until now, the use of alteplase has been limited to research or "off label" therapy in most of Europe. Despite the growing importance of this therapy and the many guidelines for use and comments on its effects, there is a lack of information about informed consent. The process of obtaining informed consent in the therapeutic decision process allows the patients to express their point of view on the risk-to-benefit ratio of thrombolytic therapy, on the different outcomes of stroke, and on the effect of thrombolysis. The discussion between the clinician and the patient or patient's family, during the process of obtaining consent to thrombolysis, can be problematic because therapy has to be given with extreme urgency. In this article, I highlight the importance of obtaining informed consent, help set out the process, and analyse the situations in which obtaining consent to thrombolysis may be legally and ethically avoided. PMID- 12849155 TI - A case of splanchnic steal syndrome? PMID- 12849162 TI - An artificial distinction. PMID- 12849163 TI - The truth is out there. PMID- 12849165 TI - Parke-Davis whistle-blower speaks at last. PMID- 12849170 TI - Dysfunctional cellular transport causes neurodegeneration. PMID- 12849173 TI - Take your pyridostigmine: that's an (ethical?) order! PMID- 12849174 TI - EUCARE--past, present, and future. PMID- 12849175 TI - Childhood opsoclonus myoclonus. PMID- 12849176 TI - Rational therapy for relapsing multiple sclerosis. PMID- 12849177 TI - Gulf war syndrome: narrowing the possibilities. PMID- 12849179 TI - Stroke in Papua New Guinea. PMID- 12849181 TI - DNA microarrays: translation of the genome from laboratory to clinic. AB - As the complete sequences of human and other mammalian genomes become available we are faced with the challenge of understanding how variation in sequence and gene expression contributes to neurological and psychiatric disorders. DNA microarrays, or DNA chips, provide the means to measure simultaneously where and when thousands of genes are expressed. Microarrays are changing the way that researchers approach work at the bench and have already yielded new insights into brain tumours, multiple sclerosis, acute neurological insults such as stroke and seizures, and schizophrenia. The study of disease-related changes in gene expression is the first step in the long process in translation of genome research to the clinic. Eventually, the changes observed in microarray studies will need to be independently confirmed and we wil need to understand how gene expression changes translate into functional effects at the cellular level in the nervous system. Progress in these studies will translate into array-based disease classification schemes and help optimise therapy for individual patients based on gene expression patterns or their genetic background. PMID- 12849182 TI - Ultrasound in the treatment of ischaemic stroke. AB - Intravenous alteplase (recombinant tissue plasminogen activator) has been shown to be beneficial within a short 3 h window after stroke. Ultrasound has a thrombolytic capacity that can be used for pure mechanical thrombolysis or improvement of enzyme-mediated thrombolysis. Mechanical thrombolysis with ultrasound needs high intensities at the clot (>2 W/cm2) that may have unwanted side-effects, whereas improvement of enzymatic thrombolysis can be done at the safer energy levels used in diagnostic ultrasound. Methods of improving enzymatic thrombolysis with ultrasound include intra-arterial delivery of thrombolytic agents with an ultrasound-emitting catheter and targeted and non-targeted non invasive transcranial ultra sound delivery during intravenous thrombolytic infusion. Animal and clinical studies of sonothrombolysis have shown clot lysis and accelerated recanalisation of arterial occlusion has been seen in in vitro flow models, occluded peripheral and coronary arteries, and intracerebral arteries. Controlled clinical trials to test safety management and effectiveness of both strategies are in progress. PMID- 12849183 TI - The therapeutic potential of cannabis. AB - Research of the cannabinoid system has many similarities with that of the opioid system. In both instances, studies into drug-producing plants led to the discovery of an endogenous control system with a central role in neurobiology. Few compounds have had as much positive press from patients as those of the cannabinoid system. While these claims are investigated in disorders such as multiple sclerosis spasticity and pain, basic research is discovering interesting members of this family of compounds that have previously unknown qualities, the most notable of which is the capacity for neuroprotection. Large randomised clinical trials of the better known compounds are in progress. Even if the results of these studies are not as positive as many expect them to be, that we are only just beginning to appreciate the huge therapeutic potential of this family of compounds is clear. PMID- 12849184 TI - Novel therapies for Duchenne muscular dystrophy. AB - The development of therapeutic strategies that overcome the unique problems posed by Duchenne muscular dystrophy (DMD) has lead to the development of many contemporary approaches to human disease in general. Various treatment approaches have been explored--such as pharmacological therapies and cell-based, cytokine, and genetic therapies--that are all targeted to specific features of dystrophic DMD muscle pathology. In genetic therapies, the large size of the dystrophin gene has necessitated the development and use of novel functional minidystrophin and microdystrophin genes, muscle-specific promoter systems, and gutted adenoviral systems. In addition to these well defined viral strategies, plasmid vectors and the upregulation of utrophin (a dystrophin homologue) have potential. Various novel genetic approaches--such as antisense-mediated exon skipping, gene correction, and new cytokine approaches--are also being developed. Together these exciting developments bring an effective treatment for DMD closer than ever before. PMID- 12849185 TI - Disruption of cellular transport: a common cause of neurodegeneration? AB - In many cases, the clinical manifestations of inherited neurodegenerative disorders appear after decades of normal function, which suggests that neurons may die through cumulative damage. Several genes that cause these diseases have been identified in recent years, but no common pathogenetic mechanism has been found. However, the most recent studies have begun to implicate the same mechanism in a range of neurodegenerative diseases, particularly those that involve motor neurons. The results of these studies suggest that the morphology and energy requirements of neurons make them particularly susceptible to the disruption of cellular transport systems. PMID- 12849186 TI - A misdiagnosis of multifocal motor neuropathy. PMID- 12849192 TI - Time to strike out stroke. PMID- 12849191 TI - Unstable connections. PMID- 12849194 TI - UK epilepsy plan is a "derisory response to a major issue". PMID- 12849202 TI - Mental mapping project kicks off in Japan. PMID- 12849203 TI - Approval of alteplase in Europe: will it change stroke management? PMID- 12849204 TI - Diabetes and the blood-brain barrier. PMID- 12849205 TI - Many pathways lead to hereditary spastic paraplegia. PMID- 12849206 TI - Saving the brain during coronary artery bypass grafting. PMID- 12849207 TI - Stroke and neurology: a plea from the WFN. PMID- 12849209 TI - Immunotherapy for Alzheimer's disease. AB - Recent studies in murine models of Alzheimer's disease (AD) have found that active immunisation with amyloid-beta peptide (Abeta) or passive immunisation with Abeta antibodies can lessen the severity of Abeta-induced neuritic plaque pathology through the activation of microglia. These antibodies can be detected in the serum and CSF. Whether they slow down or speed up the development and progression of AD has not been determined. Furthermore, the conditions that induce formation of such antibodies are unknown, or how specific they are to AD. However, the evidence suggests at least a potential beneficial role for some features of neuroinflammation in AD. A clinical phase II study of an active immunisation approach with AN1792 was started in 2001, but was recently suspended after some patients developed serious adverse events. These were most likely caused by the activation of the proinflammatory cascade. Immunotherapy approaches represent fascinating ways to test the amyloid hypothesis and may offer genuine opportunities to modify disease progression. This review focuses on immunisation strategies and details of the pathways involved in antibody clearance of Abeta. PMID- 12849210 TI - Genes and parkinsonism. AB - Genetic studies in families with mendelian inheritance of Parkinson's disease (PD) have reported the cloning of several disease-associated genes. These studies of rare familial forms of the disease have cast doubt on our understanding of the role of genetics in typical PD and have complicated the classification of the disorder. However, this genetic information might help us to construct a hypothesis for the pathogenetic processes that underlie PD. In this review we describe the molecular genetics of PD as currently understood to help explain the pathways that underlie neurodegeneration. PMID- 12849211 TI - Dementia associated with Parkinson's disease. AB - Dementia affects about 40% of patients with Parkinson's disease; the incidence of dementia in these patients is up to six times that in healthy people. Clinically, the prototype of dementia in PD is a dysexecutive syndrome. Loss of cholinergic, dopaminergic, and noradrenergic innervation has been suggested to be the underlying neurochemical deficits. Nigral pathology alone is probably not sufficient for the development of dementia. Although there is some controversy with regard to the site and type of pathology involved, dementia is likely to be associated with the spread of pathology to other subcortical nuclei, the limbic system, and the cerebral cortex. On the basis of more recent studies, the main pathology seems to be Lewy-body-type degeneration with associated cellular and synaptic loss in cortical and limbic structures. Alzheimer's disease-type pathology is commonly associated with dementia but less predictive. Recent evidence from small studies suggests that cholinesterase inhibitors may be effective in the treatment of dementia associated with PD. PMID- 12849212 TI - Long-term prognosis after lacunar infarction. AB - Lacunar infarcts, small deep infarcts that result from occlusion of a penetrating artery, account for about a quarter of all ischaemic strokes. These infarcts have commonly been regarded as benign vascular lesions with a favourable long-term prognosis. However, recent studies have shown that this is only the case early in the disease course. A few years after infarct, there is an increased risk of death, mainly from cardiovascular causes. The risk of recurrent stroke after lacunar infarct is similar to that for most other types of stroke, and patients have an increased risk of developing cognitive decline and dementia. Age, vascular risk factors, high nocturnal blood pressure, and severity of cerebral small-vessel disease at onset have significant prognostic implications for almost all outcomes. More studies on mechanisms, prevention, and treatment are needed to provide specific guidance on the long-term management of patients with lacunar infarcts. Risk-factor modification is likely to play a large part in therapeutic interventions targeted at this stroke subtype. PMID- 12849213 TI - Iron misregulation in the brain: a primary cause of neurodegenerative disorders. AB - High iron concentrations in the brains of patients and the discovery of mutations in the genes associated with iron metabolism in the brain suggest that iron misregulation in the brain plays a part in neuronal death in some neurodegenerative disorders, such as Alzheimer's, Parkinson's, and Huntington's diseases and Hallervorden-Spatz syndrome. Iron misregulation in the brain may have genetic and non-genetic causes. The disrupted expression or function of proteins involved in iron metabolism increases the concentration of iron in the brain. Disturbances can happen at any of several stages in iron metabolism (including uptake and release, storage, intracellular metabolism, and regulation). Increased brain iron triggers a cascade of deleterious events that lead to neurodegeneration. An understanding of the process of iron regulation in the brain, the proteins important in this process, and the effects of iron misregulation could help to treat or prevent neurodegenerative disorders. PMID- 12849214 TI - A woman with multiple sclerosis and pink saliva. PMID- 12849215 TI - Neurally-mediated complete heart block. PMID- 12849221 TI - Happiness pills. PMID- 12849222 TI - Jong-Wook Lee: new understanding, new hope? PMID- 12849224 TI - Cambridge primate research centre comes under scrutiny. PMID- 12849225 TI - Second gene identified for familial hemiplegic migraine. PMID- 12849231 TI - Melanocortins key to trigger sex on the brain. PMID- 12849232 TI - Predictive testing for Huntington's disease: a universal model? PMID- 12849233 TI - Monoamine oxidase inhibitors--is it time to up the TEMPO? PMID- 12849236 TI - Transcranial magnetic stimulation in neurology. AB - Transcranial magnetic stimulation (TMS) is a non-invasive tool for the electrical stimulation of neural tissue, including cerebral cortex, spinal roots, and cranial and peripheral nerves. TMS can be applied as single pulses of stimulation, pairs of stimuli separated by variable intervals to the same or different brain areas, or as trains of repetitive stimuli at various frequencies. Single stimuli can depolarise neurons and evoke measurable effects. Trains of stimuli (repetitive TMS) can modify excitability of the cerebral cortex at the stimulated site and also at remote areas along functional anatomical connections. TMS might provide novel insights into the pathophysiology of the neural circuitry underlying neurological and psychiatric disorders, be developed into clinically useful diagnostic and prognostic tests, and have therapeutic uses in various diseases. This potential is supported by the available studies, but more work is needed to establish the role of TMS in clinical neurology. PMID- 12849237 TI - Promotion of axonal regeneration in the injured CNS. AB - Molecules that are found in the extracellular environment at a CNS lesion site, or that are associated with myelin, inhibit axon growth. In addition, neuronal changes--such as an age-dependent reduction in concentrations of cyclic AMP- render the neuron less able to respond to axotomy by a rapid, forward, actin dependent movement. An alternative mechanism, based on the protrusive forces generated by microtubule elongation or the anterograde transport of cytoskeletal elements, may underlie a slower form of axon elongation that happens during regeneration in the mature CNS. Therapeutic approaches that restore the extracellular CNS environment or the neuron's characteristics back to a more embryonic state increase axon regeneration and improve functional recovery after injury. These advances in the understanding of regeneration in the CNS have major implications for neurorehabilitation and for the use of axonal regeneration as a therapeutic approach to disorders of the CNS such as spinal-cord injury. PMID- 12849238 TI - Familial and sporadic fatal insomnia. AB - Familial fatal insomnia (FFI)--a hereditary prion disease caused by a mutation at codon 178 of the prion-protein (PrP) gene (PRNP) that leads to a D178N substitution in the protein--and its sporadic form, sporadic fatal insomnia (SFI), have similar disease phenotypes. Both disorders have clinical features of disrupted sleep (loss of sleep spindles and slow-wave sleep and enacted dreams during rapid-eye-movement sleep), autonomic hyperactivation, and motor abnormalities (myoclonus, ataxia, dysarthria, dysphagia, and pyramidal signs). PET shows pronounced thalamic and limbic hypometabolism that becomes more widespread in later stages. Neuropathological assessment reveals severe neuronal loss and astrogliosis of the anterior medial thalamus and inferior olives, with later cerebral cortical and cerebellar involvement. Accumulation of an isoform of protease-resistant PrP fragment in FFI distinct from that found in a familial form of Creutzfeldt-Jakob disease with the same D178N mutation, shows the effect of the polymorphism at codon 129 of PRNP on phenotypic expression and the possibility of distinct prion "strains" with diverse pathological potential. Intriguing clinicopathological correlations in FFI and SFI suggest a role for the thalamolimbic system in the regulation of sleep and other circadian functions. PMID- 12849239 TI - Cardioembolic stroke: an update. AB - Embolism of cardiac origin accounts for about one fifth of ischaemic strokes. Strokes due to cardioembolism are in general severe and prone to early recurrence. The risk of long term recurrence and mortality are high after a cardioembolic stroke. Cardioembolism can be reliably predicted on clinical grounds but is difficult to document. MRI, transcranial doppler, echocardiogram, Holter monitoring, and electrophysiological studies increase our ability to identify the source of cardioembolism. Non-valvular atrial fibrillation is the commonest cause of cardioembolic stroke. Despite its enormous preventive potential, continuous oral anticoagulation is prescribed for less than half of patients with atrial fibrillation who have risk factors for cardioembolism and no contraindications for anticoagulation. Alternatives to oral anticoagulation in this setting include safer and easier to use antithrombotic drugs and definitive treatment of atrial fibrillation. Available evidence does not support routine immediate anticoagulation of acute cardioembolic stroke. PMID- 12849240 TI - A 38-year-old man with a 9 month history of neurological and cognitive impairment. PMID- 12849247 TI - Dreams. PMID- 12849248 TI - Herbal medicines--primum non nocere. PMID- 12849250 TI - Another set back for fetal transplants for Parkinson's disease. PMID- 12849258 TI - MyLifeBits: holding on to the present to preserve the past. PMID- 12849259 TI - Dietary vitamin E and Parkinson's disease: something to chew on. PMID- 12849260 TI - Alexander disease: GFAP mutations unify young and old. PMID- 12849261 TI - Artificial nutrition and hydration therapy in advanced dementia. PMID- 12849262 TI - Should neurologists measure D-dimer concentrations? PMID- 12849264 TI - Computer-assisted imaging to assess brain structure in healthy and diseased brains. AB - Neuroanatomical structures may be profoundly or subtly affected by the interplay of genetic and environmental factors, age, and disease. Such effects are particularly true in healthy ageing individuals and in those who have neurodegenerative diseases. The ability to use imaging to identify structural brain changes associated with different neurodegenerative disease states would be useful for diagnosis and treatment. However, early in the progression of such diseases, neuroanatomical changes may be too mild, diffuse, or topologically complex to be detected by simple visual inspection or manually traced measurements of regions of interest. Computerised methods are being developed that can capture the extraordinary morphological variability of the human brain. These methods use mathematical models sensitive to subtle changes in the size, position, shape, and tissue characteristics of brain structures affected by neurodegenerative diseases. Neuroanatomical features can be compared within and between groups of individuals, taking into account age, sex, genetic background, and disease state, to assess the structural basis of normality and disease. In this review, we describe the strengths and limitations of algorithms of existing computer-assisted tools at the most advanced stage of development, together with available and foreseeable evidence of their usefulness at the clinical and research level. PMID- 12849265 TI - Vascular cognitive impairment. AB - Cerebrovascular disease is the second most common cause of acquired cognitive impairment and dementia and contributes to cognitive decline in the neurodegenerative dementias. The current narrow definitions of vascular dementia should be broadened to recognise the important part cerebrovascular disease plays in several cognitive disorders, including the hereditary vascular dementias, multi-infarct dementia, post-stroke dementia, subcortical ischaemic vascular disease and dementia, mild cognitive impairment, and degenerative dementias (including Alzheimer's disease, frontotemporal dementia, and dementia with Lewy bodies). Here we review the current state of scientific knowledge on the subject of vascular brain burden. Important non-cognitive features include depression, apathy, and psychosis. We propose use of the term vascular cognitive impairment, which is characterised by a specific cognitive profile involving preserved memory with impairments in attentional and executive functioning. Diagnostic criteria have been proposed for some subtypes of vascular cognitive impairment, and there is a pressing need to validate and further refine these. Clinical trials in vascular cognitive impairment are in their infancy but support the value of therapeutic interventions for symptomatic treatment. PMID- 12849266 TI - Seronegative generalised myasthenia gravis: clinical features, antibodies, and their targets. AB - Myasthenia gravis (MG) is a well-recognised disorder of neuromuscular transmission that can be diagnosed by the presence of antibodies to the acetylcholine receptor (AChR). However, some patients (about 15%) with generalised MG do not have detectable AChR antibodies. There is some evidence, however, that this "seronegative" MG is an antibody-mediated disorder. Plasma from patients with the disorder seems to contain various distinct humoral factors: IgG antibodies that reversibly inhibit AChR function; a non-IgG (possibly IgM) factor that indirectly inhibits AChR function; and an IgG antibody against the muscle-specific kinase (MuSK). The presence of antibodies against MuSK appears to define a subgroup of patients with seronegative MG who have predominantly localised, in many cases bulbar, muscle weaknesses (face, tongue, pharynx, etc) and reduced response to conventional immunosuppressive treatments. Moreover, muscle wasting may be present, which prevents complete response to these therapies. PMID- 12849267 TI - Gastrointestinal dysfunction in Parkinson's disease. AB - There is growing recognition that gastrointestinal dysfunction is common in Parkinson's disease (PD). Virtually all parts of the gastrointestinal tract can be affected, in some cases early in the disease course. Weight loss is common but poorly understood in people with PD. Dysphagia can result from dysfunction at the mouth, pharynx, and oesophagus and may predispose individuals to aspiration (accidental inhalation of food or liquid). Gastroparesis can produce various symptoms in patients with PD and may cause erratic absorption of drugs given to treat the disorder. Bowel dysfunction can consist of both slowed colonic transit with consequent reduced bowel-movement frequency, and difficulty with the act of defecation itself with excessive straining and incomplete emptying. Recognition of these gastrointestinal complications can lead to earlier and potentially more effective therapeutic intervention. PMID- 12849268 TI - Multiple sclerosis in the Japanese population. AB - Multiple sclerosis (MS) in Asian populations is characterised by the selective and severe involvement of the optic nerve and spinal cord as well as low prevalence rates. 15-40% of cases of MS in Japan are of this "opticospinal" type. This form of MS generally has a higher age at onset and a higher female to male ratio than conventional MS. Opticospinal MS is also characterised by frequent relapses, severe disability, few brain lesions visible on MRI, long lesions extending over many vertebral segments visible on spinal-cord MRI, pleocytosis and an absence of oligoclonal bands in the CSF, and a pronounced shift in the responses of T-helper-1 and T-cytotoxic-1 cells throughout relapse and remission phases. Conventional MS in Japanese people is, like MS in white people, associated with HLA-DRB1*1501, whereas opticospinal MS is associated with HLA DPB1*0501. In Japanese people born after modernisation in the 1960s, the ratio of conventional to opticospinal MS has increased rapidly. Opticospinal MS is likely to have a distinct immune-mediated mechanism, which is not operative in conventional MS. PMID- 12849269 TI - MRI of basilar-artery-aneurysm growth. PMID- 12849276 TI - Feral children. PMID- 12849277 TI - vCJD complacency--cause for alarm? PMID- 12849279 TI - WHO struggles with appeal to prevent meningitis outbreak. PMID- 12849280 TI - New website for Alzheimer's disease launched in Spain. PMID- 12849285 TI - Fighting fire with fire in autoimmune disease. PMID- 12849290 TI - Potential of statins for the treatment of multiple sclerosis. PMID- 12849291 TI - Characterisation of cognitive impairment in schizophrenia. PMID- 12849292 TI - Homocysteine and the brain: vascular risk factor or neurotoxin? PMID- 12849293 TI - Treatment of patients with aneurysmal subarachnoid haemorrhage. PMID- 12849294 TI - Is it time to conclude that NMDA antagonists have failed? PMID- 12849297 TI - Mild cognitive impairment: prevalence, prognosis, aetiology, and treatment. AB - Mild cognitive impairment (MCI) is a recently described syndrome that is currently thought of as a transition phase between healthy cognitive ageing and dementia. Although this notion seems to be reasonable, the general nature of the term MCI--including its many definitions--makes accurate accounting of the prevalence, prognosis, and potential benefit from treatment somewhat difficult. The differences in cognitive profile and clinical progression among individuals with MCI are generally recognised. However, recent evidence also suggests that the aetiological heterogeneity among individuals with MCI could be greater than previously reported. For example, cerebrovascular disease seems to be underestimated as a potential cause of MCI. In this review, I attempt to recognise workable definitions of MCI to discuss the prevalence, pathophysiology, prognosis, and possibilities for treatment of this disorder. PMID- 12849298 TI - Immunotherapy in autoimmune neuromuscular disorders. AB - Important progress has been made in our understanding of the cellular and molecular processes underlying autoimmune neuromuscular diseases that has led us to identify targets for rational therapeutic intervention. Although antigen specific immunotherapy is not yet available, old and new immunomodulatory treatments, alone or in combination, provide effective immunotherapy for most autoimmune disorders. In parallel, the achievements of molecular medicine provide more specific yet largely experimental therapeutic tools that need to be tested in the human diseases. Here we review the principles and targets of immunotherapy for autoimmune neuromuscular disorders, address applications and practical guidelines, and give an outlook on future developments. PMID- 12849299 TI - Cellular biology of epileptogenesis. AB - The ionic currents that underlie the mechanisms of epileptogenesis have been systematically characterised in different experimental preparations. The recent elucidation of the molecular structures of most membrane channels and receptors has enabled structure-function analyses in both physiological and pathophysiological conditions. The neurophysiological and biomolecular features of epileptogenic mechanisms that putatively account for human epilepsies are summarised in this review. Particular emphasis is given to epilepsies that are associated with genetically determined alterations of ligand-gated and voltage gated ion channels. Changes in ionic currents that flow through sodium, potassium, and calcium channels can lead to different types of epilepsies. Inherited or acquired changes that alter the function of receptors for acetylcholine, glutamate, and gamma-aminobutryic acid are also involved. better understanding of the role of these epileptogenic mechanisms will promote new advances in the development of selective and targeted antiepileptic drugs. PMID- 12849300 TI - Stroke epidemiology: a review of population-based studies of incidence, prevalence, and case-fatality in the late 20th century. AB - This overview of population-based studies of incidence, prevalence, mortality, and case-fatality of stroke was based on studies from 1990. Incidence (first stroke in an individual's lifetime) and prevalence were computed by age, sex, and stroke type. Age-standardised incidence and prevalence with the corresponding 95% CI were plotted for each study to facilitate comparisons. The review shows that the burden of stroke is high and is likely to increase in future decades as a result of demographic and epidemiological transitions in populations. The main features of stroke epidemiology include modest geographical variation in incidence, prevalence, and case-fatality among the--predominantly white- populations studied so far, and a stabilisation or reversal in the declining secular trends in the pre-1990s rates, especially in older people. However, further research that uses the best possible methods to study the incidence, risk factors, and outcome of stroke are urgently needed in other populations of the world, especially in less developed countries where the risk of stroke is high, lifestyles are changing rapidly, and population restructuring is occurring. PMID- 12849301 TI - Design of future acute-stroke treatment trials. AB - Very few trials of acute stroke treatments show efficacy of a tested agent on the prespecified primary outcome. We can learn many lessons from the studies that achieve only neutral results. Preclinical studies have been flawed by use of models of transient not permanent brain ischaemia, treatments that aim to protect cerebral grey matter independently of white matter, delivery of the study drug within too short a time window after ischaemic insult, use of surrogate outcome measures in the short term instead of function in the long-term, and small sample sizes. Clinical trials have been hampered by heterogeneity in causes of stroke and inability to classify subtypes of cause; the short time available to rescue ischaemic brain tissue; the haemorrhagic transformation that can cause severe functional consequences seen frequently in infarcted brain tissue; the lack of valid, reliable, sensitive, and simple tools for assessment of functional outcome; and, above all, small treatment effects that are difficult to detect or refute. In this review, we look at the designs and results of all controlled trials of treatments for ischaemic stroke, and try to identify opportunities to improve future treatment assessment. PMID- 12849302 TI - The vegetarian who ate a sausage with curry sauce. PMID- 12849308 TI - Free will. PMID- 12849309 TI - Chemical weapons--shifting the goal posts. PMID- 12849311 TI - World Health Report charts way forward on CV disease prevention. PMID- 12849315 TI - Polymer implant promotes axon growth in rat model of SCI. PMID- 12849319 TI - Comprehensive genetic screening program started for Spanish patients with AD. PMID- 12849323 TI - Lorenzo's oil may help to prevent ALD symptoms. PMID- 12849324 TI - Memantine offers new possibilities for the treatment of VaD. PMID- 12849325 TI - Apparently sporadic CJD and covert health-care transmissions. PMID- 12849326 TI - Yet another spinocerebellar ataxia--will it ever end? PMID- 12849327 TI - Dyskinesias after transplantation in Parkinson's disease. PMID- 12849328 TI - Practice effects on cognitive tasks: a major problem? PMID- 12849329 TI - Sumatriptan versus eletriptan: which is best? PMID- 12849330 TI - A neurohistory tour in Vienna. PMID- 12849332 TI - Vagus-nerve stimulation for the treatment of epilepsy. AB - Vagus-nerve stimulation (VNS) is now an accepted treatment for patients with refractory epilepsy. There have been many studies suggesting that VNS affects the brain in such areas as the thalamus and other limbic structures. In addition, there is some evidence that norepinephrine is important in the prophylactic antiseizure effects of VNS. The efficacy of VNS has been established for partial seizure types, even in refractory patients who did not respond to surgical treatment for epilepsy. There are also data, from open-label studies, that suggest efficacy in other seizure types. Therefore, VNS seems to be a broad spectrum treatment for epilepsy. Improvement is not immediate but increases over 18-24 months of treatment. Most studies report subjective improvements in various quality-of-life measurements during treatment with VNS--objective trials have confirmed this observation. Side-effects are mainly stimulation related and reversible and they tend to decrease over time. They are generally mild to moderate and seldom necessitate the removal of the device. No idiosyncratic side effects have been reported in 12 years of experience, and VNS does not interact with antiepileptic drugs. Most adverse events are predictable and related to the specific stimulation regimen. VNS does not have cognitive and systemic side effects and can, therefore, be a valuable treatment approach even for patients who have poor tolerance of antiepileptic drugs. PMID- 12849333 TI - The genetic basis of dyslexia. AB - Dyslexia, a disorder of reading and spelling, is a heterogeneous neurological syndrome with a complex genetic and environmental aetiology. People with dyslexia differ in their individual profiles across a range of cognitive, physiological, and behavioural measures related to reading disability. Some or all of the subtypes of dyslexia might have partly or wholly distinct genetic causes. An understanding of the role of genetics in dyslexia could help to diagnose and treat susceptible children more effectively and rapidly than is currently possible and in ways that account for their individual disabilities. This knowledge will also give new insights into the neurobiology of reading and language cognition. Genetic linkage analysis has identified regions of the genome that might harbour inherited variants that cause reading disability. In particular, loci on chromosomes 6 and 18 have shown strong and replicable effects on reading abilities. These genomic regions contain tens or hundreds of candidate genes, and studies aimed at the identification of the specific causal genetic variants are underway. PMID- 12849334 TI - Intravenous immunoglobulin for chronic inflammatory demyelinating polyradiculoneuropathy: a systematic review. AB - This review discusses the efficacy and safety in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) of intravenous immunoglobulin and compares this treatment with plasma exchange and prednisolone. We searched publications from 1985 onwards for randomised controlled studies examining the effects of intravenous immunoglobulin in patients with this immune-mediated neuromuscular disorder. Six trials, with 170 patients in total, were judged eligible. A significantly higher proportion of patients improved in disability within a month after the start of treatment with intravenous immunoglobulin than with placebo (relative risk 3.17 [95% CI 1.74 to 5.75]). During this period, intravenous immunoglobulin has similar efficacy to plasma exchange and oral prednisolone; therefore which of these treatments should be the first choice is currently uncertain. An algorithm on treatment approaches for CIDP is proposed. PMID- 12849335 TI - Inflammation in multiple sclerosis: the good, the bad, and the complex. AB - Inflammation has always been thought of as detrimental in the pathophysiology of multiple sclerosis (MS). However, emerging genetic data, magnetic-resonance imaging studies, and immunopathological evidence challenge this simplistic view. The evidence leads to the conclusion that inflammation is tightly regulated, and that its net effect may be beneficial in MS, thus explaining some of the results from recent trials of anti-inflammatory agents. We argue that the use of anti inflammatory drugs to treat MS may not be appropriate in all cases. Precise identification of the inflammatory pathways to be targeted in the different phases of the disease and the timing of such interventions are therefore crucial. PMID- 12849336 TI - History of headache in medieval Persian medicine. AB - Despite the many studies into the history of headache, the ways in which the disorder was treated in medieval Persia are not well known in the west. Several documents still exist from which the definitions and treatments of headache in medieval Persia can be ascertained. These documents give detailed and precise clinical information on different types of headache. The medieval doctors listed various signs and symptoms, apparent causes, and hygienic and dietary rules for prevention of headaches. The medieval writings are both accurate and vivid, and they provide long lists of substances used in the treatment of headaches. Many of the approaches of physicians in medieval Persia are accepted today; however, still more of them could be of use to modern medicine. The main objective of this paper is to review the clinical approaches to headache used by practitioners in medieval Persia. PMID- 12849337 TI - Beware of optic neuritis! PMID- 12849338 TI - Ataxic form of central pontine myelinolysis. PMID- 12849344 TI - Dazed and confused. PMID- 12849345 TI - No sex please, we're British (neurologists). PMID- 12849347 TI - US Alzheimer's disease prevention study under attack. PMID- 12849349 TI - Ethical guidence given on genetics of normal human behaviour. PMID- 12849357 TI - Cyclo-oxygenase inhibitors and Alzheimer's: are we well ADAPTed? PMID- 12849358 TI - Anticoagulants and antiplatelet agents in acute ischaemic stroke. PMID- 12849359 TI - Refined analysis of transcranial Doppler HITS. PMID- 12849360 TI - A neurological basis of stuttering? PMID- 12849362 TI - Paraneoplastic neurological syndromes. PMID- 12849363 TI - Secretases as targets for the treatment of Alzheimer's disease: the prospects. AB - The amyloid hypothesis is still used to explain the pathogenesis of Alzheimer's disease. Despite all progress made, however, the molecular causes of the amyloid pathology, and of the tau pathology, tend to be ignored in most patients with this disorder (sporadic, late-onset). Mutant genes for amyloid precursor protein (APP) or presenilin cause early-onset familial Alzheimer's disease (<1% of all cases) and have helped to elucidate APP processing and amyloid-peptide formation by alpha, beta, and gamma secretases. Inhibition of production of amyloid peptides by inhibitors of beta and gamma secretases has been suggested as the rational and most specific therapeutic approach. Alternatively, or additionally, the activation of alpha secretase would increase non-amyloidogenic processing of APP. Here we review fundamental, genetic, and clinical arguments on which the therapeutic strategies for design of secretase agonists and antagonists are based, with special attention to physiological model systems to assess the potential of current efforts. PMID- 12849364 TI - Neuroimaging, the ischaemic penumbra, and selection of patients for acute stroke therapy. AB - Advances in neuroimaging have been central to the expansion of knowledge in the neurosciences over the past 20 years. One of the most important roles of brain imaging is in the selection of patients for acute stroke therapy. Currently, computed tomography (CT) is commonly used to select patients who have had strokes for thrombolytic therapy on the basis of the absence of haemorrhage and, more controversially, the presence of early CT changes of ischaemia. Since patients with ischaemic penumbra are more likely than those without to respond to therapy, identification of patients with this feature will become increasingly important. Although several imaging modalities can identify the penumbra, the most practical is magnetic resonance imaging (MRI) showing perfusion-weighted and diffusion weighted imaging mismatch. Although uncertainties in image interpretation remain, surrogate MRI outcome measures are becoming an important component of translational research. Future developments in imaging technologies may provide other opportunities for surrogate outcome studies. PMID- 12849365 TI - Subcortical ischaemic vascular dementia. AB - Vascular dementia is the second most common type of dementia. The subcortical ischaemic form (SIVD) frequently causes cognitive impairment and dementia in elderly people. SIVD results from small-vessel disease, which produces either arteriolar occlusion and lacunes or widespread incomplete infarction of white matter due to critical stenosis of medullary arterioles and hypoperfusion (Binswanger's disease). Symptoms include motor and cognitive dysexecutive slowing, forgetfulness, dysarthria, mood changes, urinary symptoms, and short stepped gait. These manifestations probably result from ischaemic interruption of parallel circuits from the prefrontal cortex to the basal ganglia and corresponding thalamocortical connections. Brain imaging (computed tomography and magnetic resonance imaging) is essential for correct diagnosis. The main risk factors are advanced age, hypertension, diabetes, smoking, hyperhomocysteinaemia, hyperfibrinogenaemia, and other conditions that can cause brain hypoperfusion such as obstructive sleep apnoea, congestive heart failure, cardiac arrhythmias, and orthostatic hypotension. Cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL)and some forms of cerebral amyloid angiopathy have a genetic basis. Treatment is symptomatic and prevention requires control of treatable risk factors. PMID- 12849366 TI - The hypothalamus in episodic brain disorders. AB - Episodic brain disorders (EBD) form an intriguing group of neurological diseases in which at least some of the symptoms occur in attacks. The hypothalamus integrates many brain functions, including endocrine and autonomic control, and governs various body rhythms. It seems a likely site in which the initiation of attacks of EBD can be modulated. Indeed, the hypothalamus has a crucial role in EBD such as narcolepsy and cluster headache. The same may be true for migraine and depression. Here we summarise the evidence supporting an important role for the hypothalamus in the initiation of disease episodes in various EBD. Study of the various pathophysiological concepts of EBD within the context of the hypothalamus may prove a fruitful example of cross-fertilisation between various research areas. PMID- 12849367 TI - Santiago Ramon y Cajal and the Spanish school of neurology. AB - Advances in neurology are now possible thanks to the endeavours of a few scientists who in the past laid firm foundations for the study of the nervous system. Santiago Ramon y Cajal (1852-1934) was one such pioneer of brain exploration and is acknowledged as the founder of modern neuroscience. He described the structure and organisation of virtually all parts of the nervous system and developed theories, including the neuron doctrine and the law of functional polarisation, that are the cornerstones of neuroscience. In addition to devoting his life to research, Ramon y Cajal was a dedicated teacher and mentor and created a school that greatly contributed to the flourishing of neurology. PMID- 12849376 TI - No pictures please. PMID- 12849377 TI - Mental health act or public order bill--cause for concern. PMID- 12849380 TI - Controversy over brain surgery for heroin addiction in Russia. PMID- 12849391 TI - Celebrity appeals for research dollars "intensifying" in USA. PMID- 12849392 TI - Global democratic consensus on neuropathological disease criteria. PMID- 12849393 TI - Stroke care in the UK--is it good enough? PMID- 12849394 TI - Do dietary antioxidants prevent Alzheimer's disease? PMID- 12849395 TI - The neurology of mitochondrial DNA disease. AB - In this era of "the gene and the genome", communication of complex genetic information to individuals and their families is becoming an increasingly common but difficult task for the clinician. This problem is particularly evident in the rapidly evolving field of mitochondrial disease: the clinician is faced with a diversity of clinical presentations and myriad mutations with, for many, only a loose relation between genotype and phenotype. The aim of this review is to familiarise the clinician with the main clinical syndromes encountered in practice, and to provide an overview of current concepts of mitochondrial genetics, including recent advances in molecular aetiology. In addition, we have included clinical guidance on the investigation and management of patients with suspected or proven mitochondrial disease based on our own experience over the past decade. PMID- 12849396 TI - Epilepsy in autism. AB - There is an increased but variable risk of epilepsy in autism. Three main factors -age, cognitive level, and type of language disorder--account for variability in the reported prevalence of epilepsy. The prevalence is highest in studies that have included adolescents and young adults, individuals with moderate to severe mental retardation and those with motor deficits, and individuals with severe receptive language deficits. The association of autism with clinical or subclinical epilepsy might denote common genetic factors in some cases. Whether subclinical epilepsy has adverse effects on cognition, language, and behaviour is debated, as is the relation of autistic regression with an epileptiform electroencephalogram to Landau-Kleffner syndrome. There is no evidence-based treatment recommendation for individuals with autism, regression, and subclinical epilepsy. Double-blind studies with sufficient power to resolve this issue are urgently needed. PMID- 12849397 TI - Progressive supranuclear palsy: where are we now? AB - This review provides an update on progressive supranuclear palsy (PSP, or Steele Richardson-Olszewski disease), an adult-onset neurodegenerative disorder characterised by early postural instability, which leads to falls, and a vertical supranuclear-gaze palsy. Recent epidemiological studies have shown that the disorder is more common than previously recognised, that it is commonly misdiagnosed, and that it may present to a wide range of hospital specialists. The diagnosis of PSP hinges on clinical acumen. Attempts to identify a suitable biomarker in the CSF or a specific and sensitive imaging or neurophysiological technique have so far failed to have a significant effect on the diagnostic process. Better understanding of the molecular pathology of PSP has highlighted the importance of tau-protein accumulation and tau-genotype susceptibility in its pathogenesis. No drug treatment significantly and consistently benefits patients, and novel therapies are urgently required. PMID- 12849398 TI - Friedreich's ataxia: treatment within reach. AB - Friedreich's ataxia (FA) is the most prevalent cerebellar ataxia in children and adults in Europe. FA is one of a growing number of diseases known to be caused by triplet-repeat expansions. The causative mutation is a GAA trinucleotide-repeat expansion in the first intron of the frataxin gene. The mitochondrial localisation of frataxin and decreased oxidation activity in vivo and in vitro show that FA is a mitochondrial disease. Frataxin is involved in iron metabolism and may protect mitochondria from oxidative damage. The understanding of the disease has only just begun and possible treatments are within reach. In this review I discuss the clinical knowledge of FA and recent developments that have helped to elucidate the pathogenesis of the disease and made the first therapeutic attempts possible. PMID- 12849399 TI - When should temporal-lobe epilepsy be treated surgically? AB - Our current knowledge of mesial-temporal-lobe epilepsy (MTLE) is extensive, yet still insufficient to draw final conclusions on the optimal approach to its therapy. MTLE has been well characterised and can usually be identified with noninvasive studies including scalp electroencephalography (EEG) and video monitoring with ictal recording, magnetic resonance imaging, single-photon emission computed tomography, positron emission tomography, neuropsychological assessment, and historical and clinical data. Sometimes, invasive EEG is needed to confirm mesial-temporal-lobe seizure onset, which, combined with the underlying pathological abnormality (the substrate) of mesial temporal sclerosis (hippocampal neuronal loss and gliosis), defines MTLE. This disorder is the most common refractory partial epilepsy, and also the one most often treated surgically, because medical treatment fails in 75% of cases, and surgical treatment succeeds in a similar percentage. Despite the recent publication of the first randomised trial of surgical treatment for MTLE, questions remain about the neurological consequences of both medical and surgical treatment, the ultimate gains in quality of life parameters, and the precise predictors of success. Long term follow-up and analyses of multiple factors in large groups of contemporary patient populations will be necessary to fully answer the question, "is temporal lobe epilepsy a surgical disease?" Right now it should be considered one in most cases. PMID- 12849400 TI - Why did NMDA receptor antagonists fail clinical trials for stroke and traumatic brain injury? AB - Glutamate N-methyl-D-aspartate (NMDA) receptor antagonists (competitive receptor antagonists, ion channel blockers, and glycine antagonists)--such as selfotel, aptiganel, eliprodil, licostinel and gavestinel--failed to show efficacy in clinical trials of stroke or traumatic brain injury. This failure has been attributed to the deficient properties of the molecules that entered human trials and to inappropriate design of clinical studies. In this article we hypothesise that glutamate may be involved in the acute neurodestructive phase that occurs immediately after traumatic or ischaemic injury (excitotoxicity), but that, after this period, it assumes its normal physiological functions, which include promotion of neuronal survival. We propose that NMDA receptor antagonists failed stroke and traumatic brain injury trials in human beings because blockade of synaptic transmission mediated by NMDA receptors hinders neuronal survival. PMID- 12849401 TI - Fits and deafness. PMID- 12849402 TI - Fibroelastoma of the mitral valve--a curable cause of stroke. PMID- 12849408 TI - A no brainer. PMID- 12849409 TI - Gene patenting--diagnosing the problem. PMID- 12849411 TI - Olfactory ensheathing cells to be used in spinal-cord repair trial. PMID- 12849414 TI - Future uncertain for variant Creutzfeldt-Jakob disease. PMID- 12849416 TI - Beta-catenin, brains, and beyond. PMID- 12849421 TI - St John's wort for the treatment of depression. PMID- 12849422 TI - Pathological tau proteins in argyrophilic grain disease. PMID- 12849423 TI - The Women's Health Initiative trial and stroke risk. PMID- 12849425 TI - The potential of anti-inflammatory drugs for the treatment of Alzheimer's disease. AB - Genetic evidence suggests that generation of amyloid beta peptide is the pivotal step in the pathophysiology of Alzheimer's disease (AD). The mechanism by which this peptide induces neurodegeneration may involve inflammatory processes. Pharmacological suppression of inflammation may therefore ameliorate the neuropathology. Basic research studies provide substantial evidence that inflammatory processes present in the brains of patients with AD are destructive, and that anti-inflammatory drugs can provide protection. Furthermore, epidemiological studies suggest that anti-inflammatory drugs reduce the risk of AD. However, there is not yet any strong evidence from completed randomised controlled trials that anti-inflammatory treatment is beneficial. Large trials of glucocorticoid therapy, hydroxychloroquine, and non-steroidal anti-inflammatory drugs (NSAIDs) in the treatment of AD have so far been disappointing. Several studies, including a large primary prevention trial with NSAIDs, are still in progress. Major issues of selection of patients, drug regimen, and duration of treatment remain unresolved. PMID- 12849426 TI - The genetics of migraine. AB - The search for genes involved in the pathophysiology of migraine poses major difficulties. First, there is no objective diagnostic method to assess the status of the individuals studied. Second, migraine is a polygenic multifactorial disorder. Familial hemiplegic migraine (FHM) is the only known autosomal dominant subtype of migraine. In half the families with FHM who have been studied, there are mutations in the calcium-channel gene CACNA1A, located on chromosome 19. In other families, a locus has been mapped on chromosome 1. The role of these loci in typical migraine is still unknown. A susceptibility locus for migraine with aura has been located on chromosome 19 (but is distinct from CACNA1A) and a genome-wide linkage analysis has mapped a susceptibility locus on chromosome 4. Another locus for migraine may be on the X chromosome. Finally, many positive association studies have been published, but few have been replicated. PMID- 12849427 TI - Paraneoplastic neurological syndromes: an update on diagnosis, pathogenesis, and therapy. AB - Since the discovery of the first clinically relevant anti neuronal antibody specific for a paraneoplastic aetiology in 1985, the number of such reactivities has grown at a rate of about one per year. Clinicians can now diagnose a paraneoplastic syndrome much more easily. This ability is especially important because, typically, the neurological symptoms occur before the cancer is diagnosed. Early tumour diagnosis is essential, because effective treatment of the cancer still seems to be the most efficient treatment option for the neurological symptoms. Immuno modulatory therapy should, nevertheless, be initiated as early as possible and seems especially helpful for peripheral syndromes and limbic encephalitis. The recent fundamental advances in understanding of the autoimmune pathology of these disorders, especially the role of cytotoxic T cells, should eventually lead to more effective treatment options. PMID- 12849428 TI - New systems for delivery of drugs to the brain in neurological disease. AB - The restricted or regulated entry of most blood-borne substances into the brain has been recognised for more than a century. The blood-brain barrier (BBB) shielding function provided by endothelial cells is important in the treatment of neurological diseases because this exclusion of foreign substances also restricts entry of many potentially therapeutic agents into the brain. The recent identification of several neuroactive proteins of potential therapeutic value has highlighted the crucial need for effective and safe transcapillary delivery methods to the brain. One promising method is delivery through brain capillaries by augmentation of pinocytotic vesicles delivery systems that use this cellular mechanism are in development. Recent investigations in animal models show that large molecules of neurotherapeutic potential can be conjugated to peptidomimetic ligands, which bind to selected peptide receptors, and are then internalised and transported in small vesicles across the cytoplasmic brain capillary barrier. These conjugates have been shown to remain functionally active and effective in animal models of neurological disease. PMID- 12849429 TI - Classification and genetics of dystonia. AB - Dystonia is a syndrome characterised by sustained muscle contractions, producing twisting, repetitive, and patterned movements, or abnormal postures. The dystonic syndromes include a large group of diseases that have been classified into various aetiological categories, such as primary, dystonia-plus, heredodegenerative, and secondary. The diverse clinical features of these disorders are reflected in the traditional clinical classification based on age at onset, distribution of symptoms, and site of onset. However, with an increased awareness of the molecular and environmental causes, the classification schemes have changed to reflect different genetic forms of dystonia. To date, at least 13 dystonic syndromes have been distinguished on a genetic basis and their loci are referred to as DYT1 to DYT13. This review focuses on the molecular and phenotypic features of the hereditary dystonias, with emphasis on recent advances. PMID- 12849430 TI - Superficial siderosis 20 years after brain tumour. PMID- 12849435 TI - Being selfish. PMID- 12849436 TI - The million dollar question. PMID- 12849448 TI - Modelling the schizophrenic brain. PMID- 12849449 TI - Rehabilitation research--time for a change of focus. PMID- 12849450 TI - Galantamine for vascular dementia. PMID- 12849451 TI - Migraine's magnetic attraction. PMID- 12849452 TI - Mutations in GABA-receptor genes cause human epilepsy. PMID- 12849453 TI - The ethical limits of neuroscience. PMID- 12849454 TI - Mouse models for neurological disease. AB - The mouse has many advantages over human beings for the study of genetics, including the unique property that genetic manipulation can be routinely carried out in the mouse genome. Most importantly, mice and human beings share the same mammalian genes, have many similar biochemical pathways, and have the same diseases. In the minority of cases where these features do not apply, we can still often gain new insights into mouse and human biology. In addition to existing mouse models, several major programmes have been set up to generate new mouse models of disease. Alongside these efforts are new initiatives for the clinical, behavioural, and physiological testing of mice. Molecular genetics has had a major influence on our understanding of the causes of neurological disorders in human beings, and much of this has come from work in mice. PMID- 12849455 TI - Deep brain stimulation for Parkinson's disease: disrupting the disruption. AB - Many people are disabled by Parkinson's disease (PD) despite the drug treatments that are currently available. For these patients, neurosurgery has the potential to help restore their function. The most effective neurosurgical procedures to date use electrical stimulation--deep brain stimulation (DBS)--of small targets in the brain by use of a pacemaker-like device to deliver constant stimulation. Although these operations can produce striking results, the mechanism by which delivery of electrical stimulation to targets deep in the brain can restore function in the motor system is not clear. This type of surgery probably works by interfering with and shutting down abnormal brain activity in areas where the current is delivered, such as the thalamus, globus pallidus, or the subthalamic nucleus. With this abnormal neuronal activity neutralised, motor areas of the brain can resume their function and normal movements are reinstated. Current research is aimed at elucidating how DBS works and using this information to develop better treatments for patients with PD and other neurological disorders. PMID- 12849456 TI - The role of nitric oxide in multiple sclerosis. AB - Nitric oxide (NO) is a free radical found at higher than normal concentrations within inflammatory multiple sclerosis (MS) lesions. These high concentrations are due to the appearance of the inducible form of nitric oxide synthase (iNOS) in cells such as macrophages and astrocytes. Indeed, the concentrations of markers of NO production (eg, nitrate and nitrite) are raised in the CSF, blood, and urine of patients with MS. Circumstantial evidence suggests that NO has a role in several features of the disease, including disruption of the blood-brain barrier, oligodendrocyte injury and demyelination, axonal degeneration, and that it contributes to the loss of function by impairment of axonal conduction. However, despite these considerations, the net effect of NO production in MS is not necessarily deleterious because it also has several beneficial immunomodulatory effects. These dual effects may help to explain why iNOS inhibition has not provided reliable and encouraging results in animal models of MS, but alternative approaches based on the inhibition of superoxide production, partial sodium-channel blockade, or the replacement of lost immunomodulatory function, may prove beneficial. PMID- 12849457 TI - The genetics of sleep disorders. AB - The contribution of genetic components to the pathology of sleep disorders is increasingly recognised as important. Genetic studies have identified genes that may be important in the regulation of circadian rhythms, which in turn determine the time of sleep onset and waking. Recent studies have shown that mutations in hPER2 are associated with autosomal-dominant familial advanced-sleep-phase syndrome. Genetic studies in a canine model of narcolepsy and in knock-out mice have led to the identification of the hypothalamic hypocretin (orexin) neurotransmitter system as a key target for human narcolepsy. The contribution of genetic factors to obstructive sleep apnoea syndrome (OSAS) has led to a better understanding of this complex disorder that may be part of a larger syndrome associated with respiratory, cardiovascular, and metabolic dysfunction. The aim of this review is to discuss the current knowledge on the role of genetic factors in sleep disorders, in particular circadian disorders, narcolepsy, restless-legs syndrome, and OSAS. PMID- 12849458 TI - Pathophysiology of cluster headache: a trigeminal autonomic cephalgia. AB - Cluster headache is a form of primary neurovascular headache with the following features: severe unilateral, commonly retro-orbital, pain accompanied by restlessness or agitation, and cranial (parasympathetic) autonomic symptoms, such as lacrimation or conjunctival injection. It occurs in attacks typically of less than 3 h in length and in bouts (clusters) of a few months during which the patient has one or two attacks per day. The individual attack involves activation of the trigeminal-autonomic reflex; thus, such headaches can be broadly classified with the other trigeminal-autonomic cephalgias, such as paroxysmal hemicrania and the syndrome of short-lasting unilateral neuralgiform headache attacks with conjunctival injection and tearing. Observations of circadian biological changes and neuroendocrine disturbances have suggested a pivotal role for the hypothalamus in cluster headache. Functional neuroimaging with PET and anatomical imaging with voxel-based morphometry have identified the posterior hypothalamic grey matter as the key area for the basic defect in cluster headache. PMID- 12849459 TI - Experiences of Parkinson's disease in India. PMID- 12849465 TI - Dyscalculia. PMID- 12849466 TI - A scared new world? PMID- 12849479 TI - Multiple sclerosis exacerbations and infection. PMID- 12849480 TI - Neuroprotection after cardiac arrest. PMID- 12849481 TI - Cannabinoids on trial for multiple sclerosis. PMID- 12849483 TI - New horizons for stroke prevention: PROGRESS and HOPE. AB - Inhibitors of angiotensin-converting enzyme (ACE) act by blocking the conversion of angiotensin I to angiotensin II, which is catalysed by this enzyme. ACE inhibitors also prevent the breakdown of bradykinin, a potent vasodepressor agent, and prevent the effects of angiotensin II, which include increase in blood pressure, peripheral vasoconstriction, and stimulation of aldosterone secretion from the adrenal cortex. Physiological and pathological studies have shown that ACE inhibitors have beneficial effects, such as increasing vascular compliance, regression of periarteriolar collagen area, improvement of coronary reserve, and regression of resistance-artery structure and left-ventricular hypertrophy. Information about the role of ACE inhibitors in stroke prevention has been limited. This review explores the epidemiological evidence for hypertension as a risk factor for stroke, a national guideline for blood-pressure control to reduce the incidence of stroke and cardiovascular disease, the findings of two recently published clinical trials on prevention of stroke and cardiovascular disease after administration of ACE inhibitors (PROGRESS and HOPE), and the implications of the findings for redefinition of future management of blood-pressure control for individuals at high risk of stroke and cardiovascular disease. The PROGRESS and HOPE trials have shown that ACE inhibitors have an important role in the prevention of stroke and cardiovascular-disease events. PMID- 12849484 TI - Neurological disorders caused by inherited ion-channel mutations. AB - Several neurological diseases-including neuromuscular disorders, movement disorders, migraine, and epilepsy-are caused by inherited mutations of ion channels. The list of these "channelopathies" is expanding rapidly, as is the phenotypic range associated with each channel. At present the best understood channelopathies are those that affect muscle-fibre excitability. These channelopathies produce a range of disorders which include: periodic paralysis, myotonias, malignant hyperthermia, and congenital myasthenic syndromes. By contrast, the mechanisms of diseases caused by mutations of ion channels that are expressed in neurons are less well understood. However, as for the muscle channelopathies, a striking feature is that many neuronal channelopathies cause paroxysmal symptoms. This review summarises the clinical features of the known neurological channelopathies, within the context of the functions of the individual ion channels. PMID- 12849485 TI - After a decade of intravesical vanilloid therapy: still more questions than answers. AB - Vanilloid sensitivity is a functional signature of a subset of unmyelinated fibres innervating the urinary bladder. The role that these nerves have in the physiological control of storage and voiding is unclear. However, after the bladder has been disconnected by spinal injury from the pontine micturition centre, vanilloid-sensitive fibres assume a central role in the reflex emptying of the bladder that occurs at low volumes. Intravesical vanilloid (capsaicin or resiniferatoxin) administration is beneficial in this disorder by "desensitising" these nerves. Resiniferatoxin is superior to capsaicin in terms of its tolerability profile. Investigators are moving rapidly to identify the mechanisms by which desensitisation to vanilloids occurs. Vanilloids induce lasting, but fully reversible, changes in gene expression, including downregulation of the vanilloid receptor subtype 1. It is hoped that application of gene chip technologies will address the global profile of vanilloid-induced changes in gene expression and their relative contribution to desensitisation. Drugs that target signalling mechanisms that bring about these changes in gene expression have obvious therapeutic potential. PMID- 12849486 TI - Is epilepsy a progressive disorder? Prospects for new therapeutic approaches in temporal-lobe epilepsy. AB - During the past decade, it has become apparent that neural circuits undergo activity-dependent reorganisation. In pathological disorders with recurring episodes of excessive neural activity, such as temporal-lobe epilepsy, brain circuits can undergo continual remodelling. For clinical practice, seizure induced remodelling implies that after a diagnosis of epilepsy, recurring seizures can cause continuing neural reorganisation and potentially contribute to progressive severity of the epilepsy and to cognitive and behavioural consequences. The alterations induced by seizures include neuronal death and birth, axonal and dendritic sprouting, gliosis, molecular reorganisation of membrane and extracellular-matrix proteins, and intermediates involved in cellular homoeostasis. These changes are influenced by genetic background and seizure type, thus identification of genetic risk factors should be a priority. Therapeutic modification of seizure-induced molecular and cellular responses offers new opportunities for intervention beyond seizure suppression. PMID- 12849487 TI - Phantom-limb pain: characteristics, causes, and treatment. AB - Phantom-limb pain is a common sequela of amputation, occurring in up to 80% of people who undergo the procedure. It must be differentiated from non-painful phantom phenomena, residual-limb pain, and non-painful residual-limb phenomena. Central changes seem to be a major determinant of phantom-limb pain; however, peripheral and psychological factors may contribute to it. A comprehensive model of phantom-limb pain is presented that assigns major roles to pain occurring before the amputation and to central as well as peripheral changes related to it. So far, few mechanism-based treatments for phantom-limb pain have been proposed. Most published reports are based on anecdotal evidence. Interventions targeting central changes seem promising. The prevention of phantom-limb pain by peripheral analgesia has not yielded consistent results. Additional measures that reverse or prevent the formation of central memory processes might be more effective. PMID- 12849488 TI - Treatment of neuropathic pain in Sierra Leone. AB - During Sierra Leone's violent decade-long war, the warring parties used amputation, especially of arms, as a means of terror. In a camp for amputees in the capital city Freetown, Medecins Sans Frontieres established a clinic and a treatment programme for neuropathic pain. Insecurity and cultural and language barriers have complicated this work, but medical and humanitarian benefits have been demonstrated. Pain services are virtually non-existent in less-developed countries. There have recently been no major treatment advances for neuropathic or phantom pain; however, the general body of knowledge about amputation pain can be increased by observations from these difficult settings. PMID- 12849494 TI - Rogue peptide theories. PMID- 12849495 TI - Rabies--what can neurologists do? PMID- 12849497 TI - New surgical interventions for Parkinson's disease. PMID- 12849506 TI - Remote possibility of sensory prostheses. PMID- 12849507 TI - Emotions and the brain: linking affective disorders to brain regions. PMID- 12849508 TI - Cell therapy for Huntington's disease, the next step forward. PMID- 12849509 TI - Hydrogen magnetic resonance spectroscopy in Alzheimer's disease. PMID- 12849510 TI - Deciphering Parkinson's disease--PARK8. PMID- 12849512 TI - The placebo effect in neurological disorders. AB - Recent evidence suggests that the placebo effect is mediated by the dopaminergic reward mechanisms in the human brain and that it is related to the expectation of clinical benefit. On the basis of this theory, we propose some criteria for the proper investigation of the placebo effect, and review the evidence for a placebo effect in Parkinson's disease, depression, pain, and other neurological disorders. We also discuss the evidence for the use of placebos in long-term substitution programmes for the treatment of drug addiction. PMID- 12849513 TI - Treatment of neural injury with marrow stromal cells. AB - We describe our preclinical studies on the use of bone-marrow stromal cells (MSC; an uncharacterised mixed population of plastic-adherent cells) in the treatment of neural injury. These cells obtained from donor rats or human beings have been directly transplanted into brain or administered intra-arterially or intravenously. MSC selectively target injured tissue and promote functional recovery. Signals that target inflammatory cells to injured tissue probably direct MSC to injury sites. Although some MSC express proteins typical of neural cells, the possibility that benefit is derived by replacement of infarcted tissue with differentiated MSC is highly unlikely. MSC activate endogenous restorative responses in injured brain, which include angiogenesis, neurogenesis, and synaptogenesis. Given the robust therapeutic benefit of these cells in the treatment of experimental neural injury, and the fact that MSC have been used in the treatment of other human disease, there is justification for further preclinical studies leading to clinical trials for the treatment of neural injury such as stroke. PMID- 12849514 TI - Human rabies: a disease of complex neuropathogenetic mechanisms and diagnostic challenges. AB - Rabies is inevitably fatal and presents a horrifying clinical picture. Human rabies can manifest in either encephalitic (furious) or paralytic (dumb) forms. The brainstem is preferentially involved in both clinical forms, though there are no clinical signs of brainstem dysfunction. Differences in tropism at the inoculation site or the CNS, in the route of spread, or in the triggering of immune cascades in the brainstem may account for clinical variation. Rabies still poses diagnostic problems, particularly the paralytic form, which closely resembles Guillain-Barre syndrome, or when a patient is comatose and cardinal signs may be lacking. Molecular methods allow reliable detection of rabies-virus RNA in biological fluids or tissue before death. Deviations from the recommendations on prophylaxis of the World Health Organization lead to unnecessary loss of life. To date, attempts to treat human rabies have been unsuccessful. PMID- 12849515 TI - Hereditary motor and sensory neuropathies: a biological perspective. AB - Mutations in genes expressed in Schwann cells and the axons they ensheath cause the hereditary motor and sensory neuropathies known as Charcot-Marie-Tooth (CMT) disease. At present, mutations in ten different genes have been identified, chromosomal localisation of many other distinct inherited neuropathies has been mapped, and new genetic causes for inherited neuropathies continue to be discovered. How to keep track of these mutations is a challenge for any neurologist, partly because the mutations are commonly presented as an expanding list to be memorised without a biological context of how the encoded proteins behave in the cell. A further challenge for investigators studying diseases of the peripheral nervous system is the increasing complexity of myelination, axonal function, and interactions between Schwann cells and axons. To address these concerns, we present the mutated genes causing these inherited neuropathies in the context of the cell biology of the Schwann cell and axon, and we begin to develop a model of how the various genes may interact in the pathogenesis of CMT disease. PMID- 12849516 TI - Coronary-artery bypass surgery and the brain: persisting concerns. AB - Mortality after coronary-artery bypass surgery (CABS) has fallen steadily over recent years. Concern remains, however, about the effect of this surgery on the brain. The problem of brain damage after CABS is multifactorial, involving microembolism, disturbed perfusion, metabolic derangement, and inflammatory responses. Microemboli numbers have been linked to the likelihood of neuropsychological deterioration after surgery. Risk factors for cerebral changes after CABS include older age, gender, neurological disease, diabetes, and calcification of the aorta. These risk factors are important because, in comparison with the early 1990s, patients undergoing CABS are now older and tend to have a greater number of comorbid conditions. Changes in surgical technique, such as the introduction of arterial-line filters and membrane oxygenators, have led to a reduction of both microemboli and neuropsychological disturbance. However, the problem persists, prompting further studies on surgical technique and neuroprotective strategies. PMID- 12849517 TI - A paradigm shift in neurorehabilitation. AB - The practice of neurorehabilitation in the clinic has undergone a paradigm shift as a result of influences from basic and clinical research. I have identified six areas of knowledge that by advancing so rapidly have brought about this paradigm shift: first, the increased understanding of how the CNS is reorganised after training or injury; second, the knowledge of how declarative and procedural memory operates and how this can influence rehabilitation therapy; third, a greater appreciation of the chemical factors that promote learning and neural remodelling; fourth, the fact that computational neuroscience can teach us how complex behaviour can emerge from the interaction of thousands of neurons; fifth, the influence of evidence-based medicine on neurorehabilitation; and sixth, the importance of reliable outcome measures for both injury and treatment. These are young scientific disciplines that offer great opportunities for further research. The complexity of neurorehabilitation will also require greater attention to a substantially neglected problem, the incorporation of techniques that have been proven effective in clinical trials into routine and effective clinical practice. PMID- 12849518 TI - Conn's dementia. PMID- 12849524 TI - Tetrachromats. PMID- 12849527 TI - Dosing in phase II trial of Alzheimer's vaccine suspended. PMID- 12849538 TI - Predictive value of MRI findings in multiple sclerosis. PMID- 12849539 TI - Susceptibility testing for Alzheimer's disease: race for the future. PMID- 12849540 TI - Do leisure activities protect against Alzheimer's disease? PMID- 12849541 TI - Structural magnetic resonance imaging in the practical assessment of dementia: beyond exclusion. AB - Neuroimaging is increasingly used to aid diagnosis in dementia. The traditional view that imaging is important solely as means of excluding treatable causes of dementia is maintained by many guidelines. These conditions however, account for a tiny proportion (<1%) of all causes of dementia. Over the past few years it has been recognised that a more accurate diagnosis and prognosis is important for patients and their families. The different pathological processes that produce cerebral dysfunction at a cellular level also produce macroscopic effects that can be detected in vivo with imaging. Clinically useful measures that distinguish between neurodegenerative disorders at an early stage are still awaited. The most likely future use of structural imaging will be the identification of patients at risk for Alzheimer's disease or with preclinical Alzheimer's disease. For magnetic resonance imaging (MRI) this will mean focusing on those areas that are affected earliest in the disease; ie, entorhinal cortex and hippocampus, using high resolution structural MRI or sophisticated brain mapping techniques. Imaging research is also likely to focus on measuring progression and detecting therapeutic effect. As such, MRI is already become an indispensable tool in clinical trials in dementia. PMID- 12849542 TI - Prediction of epileptic seizures. AB - For almost 40 years, neuroscientists thought that epileptic seizures began abruptly, just a few seconds before clinical attacks. There is now mounting evidence that seizures develop minutes to hours before clinical onset. This change in thinking is based on quantitative studies of long digital intracranial electroencephalographic (EEG) recordings from patients being evaluated for epilepsy surgery. Evidence that seizures can be predicted is spread over diverse sources in medical, engineering, and patent publications. Techniques used to forecast seizures include frequency-based methods, statistical analysis of EEG signals, non-linear dynamics (chaos), and intelligent engineered systems. Advances in seizure prediction promise to give rise to implantable devices able to warn of impending seizures and to trigger therapy to prevent clinical epileptic attacks. Treatments such as electrical stimulation or focal drug infusion could be given on demand and might eliminate side-effects in some patients taking antiepileptic drugs long term. Whether closed-loop seizure prediction and treatment devices will have the profound clinical effect of their cardiological predecessors will depend on our ability to perfect these techniques. Their clinical efficacy must be validated in large-scale, prospective, controlled trials. PMID- 12849543 TI - Cell transplantation, myelin repair, and multiple sclerosis. AB - A decade ago, therapeutic strategies to remyelinate the CNS in diseases such as multiple sclerosis had much experimental appeal, but translation of laboratory success into clinical treatments appeared to be a long way off. Within the past 12 months, however, the first patients with multiple sclerosis have received intracerebral implants of autologous myelinating cells. Here we review the clinical and biological problems presented by multiple sclerosis disease processes, and the background to the development of myelin-repair strategies. We attempt to highlight those areas where difficulties have yet to be resolved, and draw on various experimental findings to speculate on how remyelinating therapies are likely to develop in the foreseeable future. PMID- 12849544 TI - Management of acute stroke. AB - Acute stroke is a medical emergency. The outcome is influenced by the rapidity and quality of initial care, which is best delivered by specialists with appropriate facilities, including 24 h access to computed tomography. Stroke management requires attention to simple details and has been summarised in clinical guidelines. General measures include control of blood glucose concentration, temperature, fluid balance, and oxygenation. Blood-pressure management is a matter for continuing research. Aspirin should be started early in ischaemic stroke, but heparin has doubtful value; antiembolism stockings may be preferable. Thrombolysis has a substantial benefit in selected circumstances, and trials continue to examine a wider role. Neuroprotection remains experimental, though magnesium is potentially of interest. Early neurosurgery has a role limited to decompression of lesions in the posterior fossa but is under trial for large hemispheric infarcts and moderate intracerebral bleeds. PMID- 12849545 TI - Growth-factor gene therapy for neurodegenerative disorders. AB - Preclinical neuroscience has advanced rapidly over the past two decades. New approaches for treating neurological disease, including gene-based therapies, nervous-system growth factors, stem cells, novel vaccines, and modulation of the immune system, offer the potential to prevent cell loss and degeneration in the brain, rather than attempting to compensate for loss after it has occurred. I will review one of these prospective therapies: growth-factor gene therapy for Alzheimer's disease, an approach that is currently the subject of a phase I clinical trial. Other disease targets for gene therapy will also be discussed, including Parkinson's disease, Huntington's disease, inborn errors of metabolism, and cancer. The progress of gene-therapy clinical trials is aiding the transition to molecular and gene-targeted therapeutic approaches which have the potential to improve dramatically the prognosis of neurological disease. PMID- 12849546 TI - A neurologist in Zambia. AB - Those uninitiated to medicine in the tropics may consider neurology and neurologists superfluous in this environment. In actual fact, the high burden of neurologic disorders combined with a dearth of physician-level care providers brings such expertise into high demand. Clinical work in sub-Saharan Africa offers the neurologist an opportunity to care for a wide range of conditions, including disorders Westerners may only be familiar with via historical accounts. Physicians starved of academic exchange and paramedical-care providers who frequently function without physician-level support are equally enthusiastic for additional training in the diagnosis and treatment of the nervous system disorders which are encountered daily. PMID- 12849547 TI - How should stroke services be organised? PMID- 12849556 TI - The first word. PMID- 12849557 TI - Mind the gap: research and practice in cancer care. PMID- 12849558 TI - Advanced prostate cancer patients' relationships with their spouses following hormonal therapy. AB - The paper analyses the adverse impact of the hormonal therapy's side-effects on advanced prostate cancer patients' capacity for maintaining their pre-treatment spousal relationships, the pros and cons of their ways of coping with the difficulties involved, and the implications of these issues for oncology nursing interventions. In-depth interviews were conducted with 15 Israeli prostate cancer patients, and the data were processed according to the constant comparative analysis method. The findings show that partners' supportive attitude and patients' attempts to stage resilience led to pre-treatment relationships' maintenance, and that, accordingly, patients' rejection by their partners and resignation to this difficulty resulted in spousal ties' disruption. However, whereas the relationships' maintenance came at the cost of burdensome constant pretence, their disruption was eventually reconstructed by the patients in positive terms. These findings are analysed against the backdrop of the constraints imposed by the hormonal therapy on patients' general coping competence and ability to enjoy non-sexual activities, which impair their capability to solve their marital problems more effectively. They would thus benefit from training oncology nurses who specialize in the unique difficulties facing them and their partners, and from oncology nursing interventions that focus on relevant information provision and counselling. PMID- 12849562 TI - The effect of cytotoxic chemotherapy on female fertility. AB - Nurses are now primarily responsible for the provision of patient information, and the administration of an escalating number of cytotoxic agents. This paper aims to provide nurses with key information concerning the adverse effects of cytotoxic chemotherapy on the reproductive system of women of childbearing age. The provision of information on gonadal function and fertility is vital, particularly with the increase in the survival of women treated for cancer, and the trend towards women starting a family later in life. Gonadal toxicity of the various cytotoxic agents, disruption of the menstrual cycle, premature menopause, avoidance of pregnancy, chemotherapy in pregnancy, and fertility prospects post chemotherapy are addressed in this paper. PMID- 12849563 TI - Quality and efficacy of educational materials on cancer-related fatigue: views of patients from two European countries. AB - Cancer-related fatigue is a symptom with great implications for the quality of life of those that experience it. It is regarded as one of the most distressing symptoms that people with cancer develop. Its aetiology is complex, and although the mechanisms underlying fatigue have not been fully clarified it is evident that it is exacerbated by treatments intended to cure or palliate the disease. Patients at risk of cancer-related fatigue need access to information that will enable them to manage it effectively. There are a growing number of materials available to patients in different European countries on this topic, but it is unclear how useful patients find these. This study was undertaken to explore this through conducting focus groups with patients in the United Kingdom and Switzerland. One focus group was conducted in each country. Findings from these determined that individuals voiced common concerns: fatigue had not been addressed in the clinical setting - individuals surmised why this occurred; participants had not accessed materials on cancer-related fatigue previously; they made recommendations for future resources for patients. However, what was evident was that unless patients can access materials on this topic, their quality becomes purely an academic issue. PMID- 12849564 TI - Establishing nursing research priorities on a paediatric haematology, oncology, immunology and infectious diseases unit: involving doctors and parents. AB - Following a Delphi survey undertaken with nurses on a specialist children's unit to identify priorities for nursing research, this paper outlines the results of a survey to ascertain the views of doctors and parents regarding the results of the Delphi. This approach was in keeping with national guidance on multi-professional working in paediatric oncology and the importance of service-user involvement in planning and evaluating care. Convenience samples of doctors (n=16) and parents (n=10) were asked to rank the priorities previously identified by nurses. Results highlighted that in the main, nurses, doctors and parents agreed on the key areas that should take priority for research. Nurse's knowledge of day-to-day symptom management, children's quality of life, negotiation and communication in relation to care provision were identified by all three groups as high priorities. There were some areas where the views varied: this was generally in relation to the different primary focus of the individual groups-parents being very concerned with the effect of daily ward routines and procedures and their child's overall hospital experience, nurses with issues such as staff retention and morale, whereas doctors were more concerned with issues around information giving and consent to treatment. Limitations of the study, including sample selection and the transient nature of the population involved are discussed within the paper. The paper concludes that all three groups shared similar views, being focused on issues directly related to patient care. Consensus between the groups should result in future research initiatives reflecting a shared focus and responding to an identified need. PMID- 12849565 TI - Emotional support for dying patients--the nurses' perspective. AB - This study describes and explains the provision of emotional support to the dying patients from the nurses' viewpoint. The results are part of a questionnaire survey concerned with terminal care on inpatient wards at community health centres in Finland. The sample consisted of 328 nurses from 32 health centres. The data were collected using a structured questionnaire in which emotional support was measured by multiple-choice items and one open-ended question. The data were analysed using content analysis. Listening, touching, expressing empathy, attending to the patients' wishes, comforting, encouraging and being present were the most common forms of emotional support. The support was mainly provided in connection with basic care. Problems concerning the provision of emotional support were focused on organisational resources, nursing staff, patients and family members. The size of the health centre, further training in the care of terminal patients and reading the literature were associated with the provision of support. In conclusion, nurses feel they need time as well as sufficient skills and competencies so that they can provide emotional support to the extent and in the way that they themselves want to. PMID- 12849566 TI - A comparison of cancer patients and nurse specialist's symptom-assessment scores, in an acute hospital. PMID- 12849567 TI - Cancer care in Belgium. PMID- 12849569 TI - Quo vantis: cancer nursing in Europe. PMID- 12849570 TI - The universal, situational, and personal needs of cancer patients and their main carers. AB - The importance of psychosocial factors in the cancer journey for both patients and their carers has been recognised in UK health policy. The aim of this paper is to consider--(1) which needs are regarded as important by patients and their carers; (2) which are the unmet needs of patients and their carers. The study focuses on the responses of 233 paired cancer patients and their carers derived from a cross-sectional questionnaire survey in three health authorities in North West England. The majority of patient-carer pairs expressed the importance of having good relationships with health-care professionals and receiving good quality information. These can be identified as "universal" needs. A minority of patient-carer pairs expressed the importance of other kinds of needs. These items were mainly to do with managing daily life, emotions, and social identity and can be identified as "situational" or "personal" needs. Where there were discrepancies between patients and carers, patients were likely to identify more needs as important. In contrast, carers have more unmet needs, reflecting their comparative neglect. Broadly, "universal" needs are being met, but "situational" and "personal" needs, particularly among carers, are relatively unmet and require greater attention on the part of nurses and primary care professionals. PMID- 12849571 TI - Is there still an indication for nursing patients with prolonged neutropenia in protective isolation?. An evidence-based nursing and medical study of 4 years experience for nursing patients with neutropenia without isolation. AB - Patients with severe neutropenia due to high-dose chemotherapy and/or total-body irradiation are at risk of serious infections and are frequently nursed in strict protective isolation. This is a costly procedure and results in a psychological burden for the patient and its significance has been debated for a long time. The introduction of very potent systemic antibiotics, antibiotic prophylaxis, haematopoietic growth factors and peripheral stem cell transplantation might have decreased the need for it. We performed a systematic literature review and conducted a medical/nursing guideline study. In the literature we searched especially for prospective randomised studies. Only six were found, these were prospective randomised studies and contradicted each other on the usefulness of protective isolation. In an initiative aimed at promoting evidence-based care, we conducted a combined medical and nursing guideline study consisting of three parts: (1) inventory of (inter) national guidelines; (2) analysis of potential sources of infection; and (3) follow-up study post-implementation of new guidelines. RESULTS: (1) The practices in different centres in Europe appeared to vary widely. (2) Micro-organisms spread easily, especially if hands are not adequately dried. Isolation does not prevent this. Based on these findings we decided to stop protective isolation. This change of policy was combined with a campaign for optimal hygiene and introduction of hand alcohol. (3) We monitored the incidence of febrile neutropenia, infections and use of systemic antibiotics and antifungals in a 3-year period without protective isolation and compared this with the findings in the preceding 3 years with isolation. No significant differences in infections and mortality were found. We concluded that abandoning protective isolation combined with increased hygienic measures in nursing of patients with severe neutropenia does not increase the risk of infections, but improves the quality of care and patient satisfaction and reduces costs. PMID- 12849572 TI - An overview of sperm cryopreservation services for adolescent cancer patients in the United Kingdom. AB - For young men, a consequence of surviving childhood malignancy can be iatrogenic infertility. Current health policies focus on the elimination of "post code lotteries" in cancer services. The extent to which sperm cryopreservation services for young men at risk of infertility from cancer treatment are provided and standardised within the United Kingdom and Ireland, was therefore the subject of this research. This paper draws on data from a three-stage study to describe sperm cryopreservation services for adolescent males, identifying current provision of sperm cryopreservation at the majority of United Kingdom Children's Cancer Study Group (UKCCSG) centres. In particular, the ways in which services are managed and written information provided to patients and their families was focused upon. Nurses from 18 of 22 UKCCSG centres responded to a questionnaire, six nurses from the replying centres participated in further, focused interviews. Results suggested that, during a 1 year period, approximately 118 adolescent males within the United Kingdom and Ireland could potentially have benefited from cryopreservation services. Of the responding centres, 15 offered a cryopreservation service. However, the majority (n=14) lacked consistency and co ordination in their service provision. The provision of written information to this patient group was limited and analysis revealed all was of a poor quality. Findings from the study led the researchers to conclude that all young men at risk of iatrogenic sterility from cancer treatment could benefit from the production and systematic application of national guidelines and that standardising sperm cryopreservation services would fit with current health policy. PMID- 12849573 TI - Research into practice: the reality of implementing a non-pharmacological breathlessness intervention into clinical practice. AB - The work of Corner and colleagues in the non-pharmacological management of breathlessness in patients with lung cancer has aroused considerable interest. Cancer and palliative care nurses are encouraged to incorporate the breathlessness intervention into their clinical practice but this has not always proved easy or straightforward. This paper draws on the authors' experience as lung cancer nurse specialists to explore some of the difficulties nurses in clinical practice may encounter when attempting to translate the research findings into their own areas and suggests ways these difficulties may be overcome. PMID- 12849574 TI - Keeping cancer patients informed: a challenge for nursing. AB - The aim of this study was to find out how much information cancer patients feel they get from nurses and physicians; how that information is provided; and what other sources patients use in their search for information. Also, the meaning of information was surveyed. The sample comprised 273 cancer patients. Data were collected with a questionnaire specifically developed for this research. Descriptive statistics and non-parametric tests were used for statistical analyses. The results indicated that there is still much to do when it comes to informing cancer patients. Around half of the respondents had not received enough information about the prognosis, the alternatives of treatment and the effects of cancer or treatment. The provision of written information by staff was regarded as insufficient. Leaflets of cancer and related issues were the most popular source of additional information. The length of time since diagnosis, employment status, physical condition and mood had the strongest associations with patients receiving or searching for information. Most respondents wanted information because it had a positive impact on their feelings and attitudes and it helped them to cope with their situation. PMID- 12849575 TI - Introducing a nurse biopsy role in a skin cancer clinic. PMID- 12849577 TI - Transforming cancer care with the help of information technology. PMID- 12849576 TI - Adolescent cancer--the need to evaluate current service provision in the UK. AB - This article suggests that service provision for adolescents with cancer requires further attention by health service researchers. Evidence of the care needs of this patient group remains scarce, particularly in terms of evaluations of the available care settings in the UK. The authors discuss the challenges facing the adolescent cancer patient population, and discuss the findings from a recent ethnographic evaluation of one of the first specialist UK units dedicated to the care of adolescents with cancer. The implications for further evaluative research, education and staff-related concerns are explored in the light of the findings. PMID- 12849578 TI - "Nothing is carved in stone!": uncertainty in children with cancer and their families. AB - Although more children are surviving childhood cancer, they and their families still face many new stressors and challenges. Understanding the experiences of childhood cancer in children and families is conditional upon building a sound and comprehensive knowledge base that is grounded in research. Accordingly, a longitudinal interpretive qualitative study was conducted to arrive at an understanding of children's and families' perspectives on having to experience the childhood cancer-symptom trajectory. A purposive sample of 39 children with cancer and their families were recruited into the study. Open-ended interviews and participant observation were the primary data collection methods. Data were analyzed by the constant comparative method of grounded theory and analysis of illness narratives. Findings revealed that in addition to the stress and suffering resulting from the children's multiple symptom experiences, children and their families experienced many other events or 'rough spots' that made cancer and life difficult to get through. Although the 'rough spots' were numerous and varied, they all resulted in children and their families experiencing feelings of uncertainty. This paper focuses on describing the nature of uncertainty experienced by the children with cancer and their families. Recommendations for nursing practice and research are discussed. PMID- 12849579 TI - Seventh Robert Tiffany Annual Nursing Lecture; Cancer nursing in the UK: practice, policy or just pretending? PMID- 12849580 TI - Women's perceptions of seroma and their drainage following mastectomy and axillary lymph node dissection. AB - AIM: The aim of this research was to study the effect of seroma on women's perception of daily functional and emotional status after surgical treatment for breast cancer. Furthermore, the experiences of the actual drainage procedure of seroma was studied. METHOD: The study had a prospective and comparative design. A study specific questionnaire was developed and used. Eighty-two women operated with modified radical mastectomy completed the forms. Forty-one women with seroma were compared with the 41 without seroma. The items focused on women's perceptions of daily-life situations, postoperative pain, problems with the surgical scar, preoperative information, general health, levels of anxiety, depression, psychosocial support and contact with the registered nurse in the hospital. RESULTS: Overall the perceived emotional and functional status, pain and general health did not differ between the two groups. Women with seroma contacted the registered nurse in the hospital more frequently after hospital discharge. Most women with seroma had no or little pain and anxiety during the aspiration and found the procedure fully acceptable. Practical information concerning self-care and the aspiration procedure was considered insufficient. The return visit to the nurse for wound observation was important and provided psychosocial support. CONCLUSION: Seroma and its drainage is well accepted. Patients should be better informed about the possibility of a seroma and its treatment. Staff continuity and particularly the presence of a special trained nurse for wound control and for psychosocial support are much appreciated. PMID- 12849581 TI - Ozone treatment for radiotherapy skin reactions: is there an evidence base for practice? AB - Clinical staff and researchers working together can do much to bridge the gap between research and practice. This paper reports on the practice of treating severe radiotherapy skin reactions with ozone therapy; a practice that has been in place for a number of years at a specialist oncology hospital in England and perceived to be beneficial in terms of wound healing and pain relief. A multidisciplinary team of clinical staff and researchers questioned the evidence base for this practice and a literature search revealed little support for the effectiveness of this treatment in this particular context. The views of patients receiving ozone therapy were sought and assessment forms were completed to gain objective information on the progress (or otherwise) of wound healing. While patients perceived the ozone treatment to be beneficial in terms of pain relief, it was impossible to isolate the impact of ozone alone as other preparations and treatments were also being given. Patient reports and nursing assessments did not support that ozone was effective at wound healing. A more formal evaluation of this treatment is being planned, supported by the shared governance initiative at the study site and a continued collaboration between clinical staff and researchers. PMID- 12849582 TI - Breast and cervical cancer screening among Korean American elderly women. AB - The purpose of this study was to estimate the prevalence and correlates of breast and cervical cancer screening tests among Korean American elderly women. This study examined the effects of individual socio-demographic background, acculturation level (e.g., proportion of life spent in the US, spoken English proficiency), health status and access to health care on uptake of mammography and Pap smear tests in the past 2 years. Subjects were 130 Korean American women aged 60 and older living in the Greater Metropolitan Baltimore Area. A cross sectional face-to-face interview was conducted. The 2-year prevalence was only 35% for mammography and 29% for Pap smear tests. In multiple logistic regression analysis, educational attainment, government support, multiple chronic conditions, and routine checkups were associated with having mammography. Age, acculturation and insurance were associated with having a Pap test. Taking advantage of the positive influence of identified covariates may increase the effectiveness of cancer control interventions among Korean American elderly women. PMID- 12849583 TI - Impact of the palliative care clinical nurse specialist on patients and relatives: a stakeholder evaluation. AB - Within the last decade, there has been a rapid expansion in the United Kingdom of clinical nurse specialists (CNSs), yet there has been limited evidence of their effectiveness. The clinical area of palliative care has one of the largest numbers of CNSs which has particularly started to escalate in the acute hospital setting. Following the Calman-Hine Report and the more recent NHS Cancer Plan, it is anticipated that the number of CNSs in palliative and cancer care is likely to grow. In light of the current policy emphasis of Clinical Governance and Clinical Effectiveness, there is an urgent need to demonstrate the contribution that CNSs make. This paper presents the qualitative findings of an evaluation study on the impact of the CNS within a palliative care team in a large acute hospital in the North-West of England. A stakeholder evaluation that encompassed tape-recorded semi-structured interviews to explore how the stakeholders saw the impact of the CNS within palliative care was performed. Twenty-seven interviews were conducted with stakeholders across the hospital setting including senior nurses, consultants, junior doctors and nurses representing the different grades. Data were analysed for emerging themes utilizing a case and cross-case analysis. The results indicated that the CNSs within palliative care in an acute hospital are seen to be potentially beneficial for both patients and relatives. This paper will focus on exploring these perceived benefits, which included symptom control, psychological care and being an advocate for patients and relatives. PMID- 12849585 TI - The worth of qualitative research. PMID- 12849584 TI - Existential concerns in late stage cancer. AB - In health care, it is generally acknowledged that individuals experiencing cancer illness and dying of cancer face a challenging time. Oncology and palliative care have responded by developing an expertise of treatment and care. The opportunity to receive the best of health care is accepted as an important concern to patients. However, recent research explains that many areas of dying cancer patients' concerns remain undisclosed and unresolved (Heaven & Maguire 1997). The purpose of this qualitative study was to identify dying cancer patients' concerns with the aim of developing a Concerns Questionnaire to further explore the relationship between patients' concerns and psychological and physical distress. In-depth interviews were carried out with 21 patients with advanced cancer who had been told they were no longer to receive curative treatments. Content analysis of the interview transcripts showed that existential concerns were a significant issue for these patients. Furthermore, patients' opportunity for reflection, personal change and inner discovery at the end of their illness could be enhanced by earlier experiences of holistic cancer care. PMID- 12849586 TI - A Delphi study on the research priorities of European oncology nurses. AB - The importance of establishing priorities for cancer nursing research has been repeatedly acknowledged (Corner 1993, Hunt 2001). A Delphi survey designed to capture research priorities of European cancer nurses was undertaken to document issues perceived as important to European Oncology Nursing Society (EONS) members and to set priorities to underpin an evolving research strategy. The survey comprised three phases, beginning at the second EONS Spring Convention (2000) when European cancer nurses were approached to complete a phase 1 questionnaire. In phase 2, research priority categories generated from responses to the phase 1 questionnaire were reviewed by a group of European cancer nurses. The categories and the placement of all the research questions within these categories were comprehensively reviewed. The final phase involved the distribution of the second questionnaire. Two hundred and twenty three nurses responded to the first questionnaire and 117 responded to the second questionnaire. Prioritisation of research categories reflected diverse views between respondents, with many categories receiving similar overall scores, of almost equal priority. Nonetheless, a number of categories demonstrated high priority throughout Europe namely needs of patients related to communication, information and education; symptom management; experiences of disease and its treatment; cancer nursing research issues (e.g. research facilitation) and cancer nursing education issues. It is recommended that future research examines clusters of research questions around the highest ranked priorities. This may provide both direction and focus to European-wide cancer nursing research, whilst ensuring clinically relevant investigation. PMID- 12849589 TI - Critical path analysis in head and neck cancer: a management technique for surgical oncology. AB - This paper describes a study of critical path analysis (CPA) as applied to the process of care for patients with head and neck cancer. CPA is a management technique, which has been developed in both the Armed Forces and a wide range of industries. It identifies the parts of a process where improvements are most likely to make an impact on the overall time for that process. There is evidence that reducing the overall times for an oncology care process has a major impact on outcome for patients. This paper describes an appraisal of a formal application of CPA in the setting of oncology. The technique was applied to a single head and neck unit. The results were compared with published standards for times in the care process and with the comparable times for the whole of the South and West of England. The critical path (the combination of processes that limits the total time taken for the whole process) at this unit ran through the combined oncology clinic, leading to a decision to increase the frequency of these clinics, which lead to a reduction in overall process time for patients. A randomised controlled trial in five centres in the South and West region will follow which includes patient quality of life outcomes and collaborative practice to assess the impact of CPA on the patient care process. PMID- 12849590 TI - The meaning of the breast cancer follow-up experience for the women who attend. AB - Attendance at a breast cancer follow-up clinic is a standard part of the after care offered to women who have had breast cancer. This research was undertaken to elicit an insight into the experience of attending for follow-up after primary breast cancer. The research was undertaken using a qualitative approach using the method of phenomenological enquiry in order to gain understanding from the perspective of the women who attend follow-up clinic. Six women were interviewed. Interviews were taped and transcribed verbatim. Analysis of the transcripts indicated a cycle of emotions associated with attendance at follow-up clinic. Required attendance produced anxiety in women who were otherwise living free from anxiety about breast cancer recurrence. This anxiety appeared to create a need to attend in order to gain reassurance of continued well-being. Despite gaining reassurance all of the women in this study reported feelings of dissatisfaction with the follow-up clinic experience. In the current climate of over-extended and under-staffed clinics it may be timely to examine current practice for effectiveness in terms of physical and psychological impact on the patient and efficient use of healthcare resources. PMID- 12849591 TI - BRCA1 and BRCA2 genetic testing for ovarian cancer: is it all good news? AB - In England and Wales, ovarian cancer is responsible for about 15 deaths per 100,000 women annually (NHS Executive 1999). It is estimated that only 5-10% of ovarian cancer cases have a hereditary basis (Watson et al. 1996, Hallowell 1999, Eeles & Powles 2000), although recent media excitement may have led some people to believe the odds are higher. The majority of these hereditary cases are thought to be caused by germline mutations in BRCA1 and BRCA2 cancer susceptibility genes (Antoniou et al. 2000, Berchuk et al. 1999). This article will examine current knowledge about BRCA1 and BRCA2 genetic testing for ovarian cancer and follow the journey taken by women who are considering such testing. Genetic counselling and genetic testing will be critically evaluated and the implications for clinical practice, the individual and society as a whole will be discussed. PMID- 12849592 TI - A survey of nurse specialists working with patients with lung cancer. PMID- 12849593 TI - Survey of information leaflets on advice for acute radiation skin reactions in UK radiotherapy centres: a rationale for a systematic review of the literature. PMID- 12849595 TI - Integrated medicine: is self-help important? PMID- 12849594 TI - Re: Management of acute radiotherapy induced skin reactions: a literature review. Eur J Oncol Nurs. 2001 Dec;5(4):221-33. PMID- 12849596 TI - An oral care protocol intervention to prevent chemotherapy-induced oral mucositis in paediatric cancer patients: a pilot study. AB - The likelihood of paediatric cancer patients experiencing oral mucositis has increased significantly as high-dose and multiple chemotherapy agents are used in the treatment of childhood cancer in recent years. The resulting oral ulcerative lesions can cause significant pain, dysphagia, alteration in nutritional status and increased risk for localized infections that could disseminate systemically. The purpose of this pilot study was to determine the clinical benefits of an oral care protocol intervention on the prevention and reduction of the severity of oral mucositis in paediatric patients receiving chemotherapy. Fourteen children were enrolled in the study; there were seven children in the control and seven in the experimental group. In the experimental group, children received a preventive oral care protocol consisting of tooth brushing, normal saline rinse and 0.2% chlorhexidine mouth rinse. Children in the control group received usual care according to the study's clinical setting. Data were collected at baseline, then twice a week for 3 weeks. The incidence of ulcerative lesions, severity of oral mucositis and the related pain intensity were used as the main outcome variables. The experimental group exhibited fewer and less painful oral mucositis lesions. The results of this study support the preventive use of oral care protocols in paediatric patients undergoing chemotherapy for cancer treatment. PMID- 12849597 TI - Helping children speak up in the health service. AB - This paper reports the main findings of a study undertaken to investigate children's perceptions of health-care professionals, the environment in which they are cared for and their information needs when sick. The main focus of the investigation was 6-10-year-old chronically sick children (predominantly with cancer). A group of healthy school children were used as a comparison in order to highlight similarities and differences. The draw and write technique and an open ended stimulus task were presented to a total of 99 children (50 hospital, 49 community). Trent NHS Executive and the South Bank University funded the project through Action for Sick Children (formerly NAWCH). The purpose of this paper is to describe how children appear to view care environments including comments on the sorts of information they want and from whom this might be obtained. A wealth of valuable qualitative data, in the form of pictures and comments were collected and are discussed here alongside a selection of the pictures. The researchers also make certain recommendations for the care of sick children, which were drawn directly from their pictures, responses and comments. A previous paper evaluated the draw and write technique and open-ended stimulus task as a tool for gaining information from a child's perspective and as a possible aid to the construction of audit tools (Bradding & Horstman 1999). A full report of this study, published by South Bank University, is available from the authors. This detailed report includes an extended range of drawings, which can be used for teaching and discussion purposes. PMID- 12849598 TI - Patients' cancer journeys in Kintyre--a qualitative study of the care, support and information needs of people with cancer and their carers. AB - Although only a short flight from Glasgow, the Kintyre area of Argyll, Scotland is 4 hours by road. Patients and their health providers face challenges when gaining access to cancer care services. As part of a Scottish government funded project, examining current services for patients with cancer, an independent qualitative study was commissioned. The method comprised selecting 30 individuals who had received cancer services within the past 3 years. Using semi-structured interviews, critical incident techniques prompts the researcher recorded patients' stories. Two main forms of information were discovered. Firstly, insight was gained into practical issues. These included how airport, health and other services operated. Secondly, the study discovered the importance of other factors in patients' lives while they were undergoing treatment. For example, patients' experiences if they had relatives living near the Glasgow treatment centres. The simple methods used in this study are generalisable in several settings particularly in communities where local factors may influence the experience of people receiving services from providers situated in a geographically distant location. Perhaps the most important feature, agreed by all parties, was that the insights gained from this study may not have been gained using more 'mainstream' quantitative methods. PMID- 12849599 TI - Breast cancer: reactions of Israeli men to their wives' diagnosis. AB - The purpose of this research was to describe Israeli husbands' reactions to their wives' diagnosis of breast cancer. This quantitative, descriptive, preliminary study employed a convenience sample of 20 men and investigated their perception of the information they received, their psychosocial responses, the effects on their marital relationship, and their perceived spousal social support. The study instruments included the Locke Wallace Marital Adjustment Scale, the Psychosocial Adjustment to Illness Scale, and a Social Support Questionnaire. This subject has not been researched in Israel previously. Husbands voiced total trust in the health-care delivery system, even though they reported a need for more information on the disease, treatment, and issues concerning their marital relationship. Husbands had difficulties with their day-to-day activities, and almost half of them cited problems both at work and at home. They described stress, including depression and sleep disturbances. They reported changes in their sexual interest and sexual functioning as well. In general, husbands felt the support of their wives. Husbands whose wives have breast cancer need a network of support to address their specific issues and concerns. The Breast Care Clinical Nurse Specialist is cited as being a pivotal component of such a support network. PMID- 12849600 TI - Evaluation of advanced home care (AHC). The next-of-kin's experiences. AB - This project studied next-of-kin's experiences of advanced home care (AHC) during the palliative care and death of a family member. The aim of the study was to evaluate how next-of-kin experienced information provided, care, symptom relief, and care-giving burden. A self-questionnaire with 24 questions was sent to all next-of-kin (n=82) who had had a family member cared for at home during one year. Seventy-five (91%) responded. Next-of-kin were generally more satisfied with emotional support and care than with information provided. Symptom relief was sufficient in most cases. Next-of-kin's experiences of burden were: 45% felt homebound, 26% felt isolated at home and 51% reported a sleep deficit. Women felt homebound (P<0.01) to a higher degree than men. However, 87% stated that they would choose AHC again, in a similar situation. AHC provided 'quite good' symptom relief and 'good' emotional support to patients and their next-of-kin during the terminal phase leading to death. Information provided needs to be improved and the situation and needs of informal caregivers requires more assessment. PMID- 12849601 TI - Practice development for cancer care. PMID- 12849602 TI - Research into the workload and roles of oncology nurses within an outpatient oncology unit. AB - This project evaluated the roles and workload of Registered Nurses (RNs) within an outpatient Cancer Care Centre (CCC) of a tertiary referral hospital in Sydney, Australia. The research incorporated the two distinct areas of radiation oncology and haematology oncology. Patient throughput (incorporating clinic attendance and treatments) was known to have increased substantially since the centre opened in 1994. The nursing Full Time Equivalents (FTEs) for the CCC were determined when the centre opened with minimal change to the nursing establishment since then. Nursing staff perceived that their workload had increased substantially and that their roles had become more complex. Work sampling was conducted over a 3-month period and identified the range of nursing activities performed by the oncology outpatient nurse. The nurses were found to have a large administrative role while their nursing activities ranged from basic nursing tasks to more specialist activities including patient counseling and complex chemotherapy regimes. PMID- 12849605 TI - Minimising the use of sedation/anaesthesia in young children receiving radiotherapy through an effective play preparation programme. AB - The delivery of radiotherapy to young children is a challenging process, yet little research has been undertaken to examine practice. Many centres routinely sedate or anaesthetise young children to overcome problems relating to non adherence to treatment. This can lead to an increased risk of clinical complications, sub-optimal nutrition, sleep disruption and is time consuming and costly. This article describes how sedation can be minimised in young children through an effective play preparation programme. To examine the effectiveness of such a programme, which had been in progress for a number of years, an audit was undertaken on a paediatric oncology unit within a regional radiotherapy centre. This took place over a 5-year period (1994-1999), on all patients aged 2-5 years. Information was collated on age, tumour type, site, duration of radiotherapy, the need for involuntary immobilisation, days requiring sedation, and whether radiotherapy was given with curative or palliative intent. Outcome data presented on 63 children suggest sedation can be minimised in this age group through the implementation of an effective play preparation programme. Of 1030 treatment days, only 111 days required sedation (10.8%), over the whole age range. No general anaesthetics were given. Only 6 patients were sedated for the whole of their treatment (9.5%), with 52 patients requiring no sedation at all (82.5%). PMID- 12849606 TI - Psychosocial and economic aspects of a trial of management of mild and borderline cervical abnormalities (TOMBOLA). AB - TOMBOLA is a randomized controlled trial to determine (a) the most effective and efficient management strategy for women with borderline or mildly dyskaryotic cervical smear results, and (b) the most appropriate treatment for women with abnormalities detected following colposcopy. This paper outlines the trial's psychosocial and economic protocol, and explains how TOMBOLA can provide detailed evidence on the psychosocial and economic impact of alternative management options and follow-up. This protocol has been developed in parallel with the clinical protocol, thereby offering the opportunity for the development of a type specific outcome measure and detailed piloting of all instruments. The multi disciplinary approach to TOMBOLA allows the combination of clinical outcomes with both generic and specific psychosocial and health-related quality of life measures. The convergent validity between these measures will be assessed, with a view to developing a specific utility index for the future assessment of cervical screening modalities. PMID- 12849607 TI - Reflecting on paediatric oncology nursing practice using Benner's Helping Role as a framework to examine aspects of caring. AB - This paper uses Benner's helping domain (Benner 1984) to reflect on two aspects of paediatric oncology nursing practice. Exemplars of care will illustrate the use of therapeutic touch and bereavement skills whilst caring for two children with cancer. Children need to be given time and opportunity to express their feelings, fears and anxiety to enable them to understand and cope with their loss. They may also need support of a familiar adult and the multi-disciplinary team. In establishing a commitment to a healing relationship touch can be a valuable tool. Touch is a complex form of non-verbal communication that can be used to convey warmth, empathy and comfort. It is important that nurses are aware of both the types and meaning so it can be used therapeutically and appropriately. Paediatric oncology nurses are ideally placed to establish meaningful relationships with children who have cancer and their families but must be aware of their own limitations. To further improve care we must continue to reflect on the richness of our practical experiences and chart our expert knowledge. PMID- 12849608 TI - Evaluation of a hospice based reflexology service: a qualitative audit of patient perceptions. AB - Complementary therapies are being increasingly used in palliative care in the drive to improve patients' emotional, psychological and spiritual health, and enhance the quality of their lives. The importance of seeking the 'user' perspective when evaluating such services is becoming increasingly acknowledged. However, it is also extremely important that we elicit such perspectives in an ethically sensitive manner. This study used a simple semi-structured questionnaire to elicit the views of a convenience sample of 34 patients receiving palliative care at a specialist palliative core unit in the north of England who had completed a course of 4-6 sessions of reflexology. Patients' comments about the therapy and the service as a whole were overwhelmingly positive. They identified relaxation, relief from tension and anxiety, feelings of comfort and improved well-being as beneficial effects of their course of reflexology. Patients also spontaneously evaluated the experience holistically in terms of the wider therapeutic environment--the centre, the staff and the therapist as well as the therapy itself. The increasing demand for evidence based practice now challenges researchers to provide a relevant holistic assessment of complementary therapies using approaches that are both ethical and sensitive to the needs of this vulnerable patient population. PMID- 12849609 TI - Institutional management of cancer-related fatigue: a comparison of clinical specialties. AB - Fatigue is the most common symptom associated with cancer and its treatment and is now widely recognized as a significant problem for patients with cancer (Ream & Richardson 1999). As a result of the restructuring of cancer services, much of the care given to patients is delivered by a multitude of nurses across a spectrum of clinical settings. While some research, albeit minimal, has evaluated nurses' knowledge and attitudes of cancer-related fatigue in various clinical settings, factors associated with institutional management of fatigue and the support and encouragement given to nurses by institutions regarding cancer related fatigue appears to be unexplored. Nurses were recruited from community, general medical, general surgical and oncology clinical settings. A postal questionnaire to evaluate institutional cancer-related fatigue management was administered. Data were analysed descriptively. Institutional management of fatigue is poor across the clinical specialties involved in this survey. Topics addressed were interdisciplinary working, documentation and standards, accountability, information giving, staff education and quality assurance initiatives. Little difference was detected across the clinical specialties for the majority of these issues. Although individual health professionals have a personal responsibility to ensure the care they provide addresses patients' needs appropriately, institutional support and direction is essential. It seems that health-care institutions have not recognized the importance of this issue. This unacceptable situation must be resolved to ensure all health professionals are supported by their institutions to make every effort to improve the management of this symptom for patients with cancer. PMID- 12849611 TI - What is HER2? AB - The focus of recent research into the cause of cancer has been the cell systems controlling growth. This has shown that mutations affecting growth factor receptors are involved in oncogenesis. The human epidermal growth factor receptor (HER) family is one of the best characterized systems in breast cancer and has important functions in the control of normal growth regulation. HER2 has a central role in the HER family, interacting with the other members of this receptor family to potentiate intracellular signalling. Although our understanding of how HER2 functions and is regulated is limited, studies have linked HER2 gene amplification/receptor overexpression to cancer development. HER2 gene amplification/receptor overexpression is observed in a significant proportion of various tumour types, occurs relatively early during cancer progression and is associated with poor prognosis in women with breast cancer. Knowing the HER2 status of patients with breast cancer will become increasingly important for making management decisions. PMID- 12849612 TI - Prognostic and predictive implications of HER2 status for breast cancer patients. AB - Women diagnosed with breast cancer face many emotional and psychological challenges, which are often related to how they perceive their chances of survival. A number of factors are known to be indicators of breast cancer prognosis, including tumour type, size and grade, and lymph node status. These factors can be used alone or in combination to make management decisions based on a broad assessment of tumour aggression. However, identifying an independent marker of prognosis has proved more difficult. The human epidermal growth factor receptor-2 (HER2) is amplified/overexpressed in 25-30% of breast tumours. This event has been shown to be associated with poor prognosis in breast cancer patients. Initial observations indicated that median survival duration is reduced by at least 50% in patients who overexpress HER2 compared with those who do not overexpress the receptor. This finding has been supported by many subsequent studies and data are accumulating to indicate that the relationship holds for both node-negative and node-positive patients. This could be particularly important for identifying the 30% of node-negative patients who will progress despite therapy. HER2 status may also be able to predict the outcome of therapy. It appears likely that HER2 overexpression predicts for resistance to tamoxifen and other hormonal therapies. It is also possible that HER2-overexpressing patients may respond better to dose-intense anthracycline therapy and worse to cyclophosphamide/methotrexate/5-fluorouracil than non-overexpressing patients. Explaining these associations to patients may empower them to deal with the psychological effects of a diagnosis of breast cancer while providing hope for the future. PMID- 12849613 TI - A practical guide to HER2 testing. AB - Nurses are becoming increasingly involved in providing information about the full range of medical care to patients. With the identification of increasing numbers of prognostic factors in cancer, this role is extending to include testing for these factors. HER2 status has prognostic and predictive value in breast cancer and the receptor is a target for specific therapies. To identify patients who might benefit from more aggressive treatment regimens or new therapeutic options, it is vital that HER2 abnormalities are accurately measured. A variety of techniques have been developed to measure HER2 overexpression or amplification, including fluorescence in situ hybridization, immunohistochemistry, polymerase chain reaction and enzyme-linked immunosorbent assay. To date, no one test has been accepted as standard and pathology laboratories are using different tests. This has implications for the interpretation of results, which in turn has implications for the widespread use of HER2 testing. This article describes the various tests available for the measurement of HER2, identifies the features of the ideal HER2 test and which test currently best fits these criteria, and considers whether HER2 status should be a routine part of the clinical evaluation of breast cancer patients. Important information that nurses should consider when providing information on HER2 testing to patients is highlighted. PMID- 12849614 TI - Herceptin: increasing survival in metastatic breast cancer. AB - Growth factors and their receptors play an important role in the pathogenesis of human cancer. The human epidermal growth factor receptor-2 (HER2) is overexpressed in approximately 30% of breast cancers and this is associated with poor clinical outcome. Overexpression of HER2 has been demonstrated to play a direct role in oncogenic transformation. Murine monoclonal antibodies (muMAbs) targeting the extracellular domain of the HER2 receptors suppress HER2-positive cancer cell growth, with muMAb 4D5 having particularly potent activity. A humanized form of muMAb 4D5 was generated by converting all but the antigen binding region of muMAb 4D5 into human IgG consensus sequences. The humanized monoclonal antibody, Herceptin, preferentially targets HER2-overexpressing cells, produces responses in breast cancer patients and is well tolerated. In a pivotal phase III trial, Herceptin administered in combination with chemotherapy (anthracycline/cyclophosphamide or paclitaxel) was compared with chemotherapy alone. The combination was found to produce significant survival benefits in HER2 positive metastatic breast cancer patients. These results have led to the approval of Herceptin for clinical use in the USA and elsewhere. PMID- 12849615 TI - Anti-HER2 therapy: how to use Herceptin in clinical practice. AB - The recombinant, humanized monoclonal antibody Herceptin is administered as an initial 4 mg/kg i.v. infusion over 90 min, followed by weekly 2 mg/kg i.v. infusions over 30 min. In pivotal clinical trials Herceptin, both as a single agent and in combination with chemotherapy, was found to produce a significant survival benefit in HER2-positive metastatic breast cancer patients. Importantly, Herceptin was well tolerated and was not associated with a poor side-effect profile, producing mainly mild to moderate side-effects. These were generally associated with the initial infusion. The incidence of adverse events was low with Herceptin therapy and severe vomiting and alopecia, typical of many chemotherapy treatment regimens, were not experienced by patients in the trials. Cardiac dysfunction was the most significant Herceptin-related adverse event but could, in most cases, be managed using standard therapy. The favourable adverse event profile seen with Herceptin is reflected in the quality of life (QoL) data. The health-related QoL of patients receiving Herceptin therapy alone or in combination with chemotherapy was found to be maintained. Previous studies have noted a deterioration of health-related QoL in patients treated with chemotherapy only. Thus, as well as providing significant survival benefit, Herceptin therapy improves patient well being. PMID- 12849616 TI - Herceptin: implications for breast cancer management. AB - It appears that no further significant improvements in the survival of women with metastatic breast cancer are likely to result from the use of traditional chemotherapeutic agents. Research attempting to identify novel agents has focused on biological therapies, which can be used to target specific abnormalities in cancer cells. Human epidermal growth factor receptor-2 (HER2) is a cell membrane receptor with growth-regulating activity. Studies indicate that the 25-30% of women with breast cancer who overexpress HER2 have aggressive disease and a worse prognosis than those who do not overexpress HER2. Herceptin targets HER2 and is the first humanized monoclonal antibody approved for therapeutic use. Clinical trials have demonstrated that Herceptin is well tolerated, produces durable objective responses and improves survival in women with metastatic breast cancer. Its side-effects are generally mild to moderate and differ from those of traditional cytotoxic agents. Cardiotoxicity, the most significant adverse effect of Herceptin, is manageable in most patients using standard therapy. The likely positioning of Herceptin in the treatment of HER2-overexpressing metastatic breast cancer is in combination with paclitaxel as first-line therapy and as second- or third-line therapy when administered alone. It is likely that the exploitation of HER2 as a target for therapy and the development of Herceptin will serve as a model for the development of future biological therapies. PMID- 12849617 TI - The challenge of gene therapy. PMID- 12849618 TI - Nurses' knowledge and attitudes towards cancer-related fatigue. AB - Fatigue is the most common symptom associated with cancer and cancer treatment and is now widely recognized as a significant problem for patients with cancer (Ream & Richardson 1999). Although the profile of cancer-related fatigue has grown over the past few years within the specialty of oncology, not all patients with cancer receive care from cancer specialists. As a result of restructuring of cancer services, the majority of patients within the UK receive their cancer treatment outwith a cancer centre and are supported by District General Hospitals and community-based services. Consequently, it is appropriate to evaluate nurses' fatigue knowledge and attitudes across a range of clinical settings caring for patients with cancer. Nurses were recruited from community, general medical, general surgical and oncology clinical settings (n=470). A postal questionnaire to evaluate fatigue knowledge and attitudes was administered. A response rate of 43% was achieved. Data were analysed descriptively. Underestimation of fatigue incidence (by 28% of respondents), poor knowledge and practice regarding fatigue assessment and management, and poor fatigue communication practices were common throughout all clinical areas. However, nurses from all settings consistently demonstrated a good understanding of the impact of fatigue on patients with cancer and an appreciation of the importance of the role of the nurse in fatigue management. While oncology nurses demonstrated the greatest fatigue knowledge, these were not significantly better than the other specialties. A significant proportion of nurses caring for patients with cancer are unaware of fatigue's incidence, assessment and management. As cancer incidence rates are set to rise and the spectrum of clinical settings in which patients receive their care is increasing, it is imperative that this common and debilitating symptom receives appropriate recognition from health-care professionals. A number of strategies to enhance knowledge and information dissemination should be initiated to ensure that patient outcomes in relation to cancer-related fatigue are improved. PMID- 12849619 TI - Management of acute radiotherapy induced skin reactions: a literature review. AB - Up to 60% of patients with cancer will receive radiotherapy treatment at some stage during their illness. One of the main side effects of this treatment is an acute skin reaction, which may range in severity from a mild erythema to moist sloughing off of the epidermis. The management of these skin reactions varies between cancer centres and is often based on personal preference and historical practices rather than clinical evidence. This paper presents a literature review that was conducted to discover the most commonly recommended treatments and the research evidence to support them. The review was conducted in two parts and looked at both clinical papers and research studies. A multitude of treatments were recommended in the clinical papers, but few were supported by high quality research. As a result of combining the evidence from both sources, preliminary guidelines for acute skin reaction management can be produced, but these should be implemented in clinical practice and their clinical performance evaluated. There is a need for further research to review treatments that have produced promising results in the reviewed research studies and to evaluate other commonly recommended topical treatments. PMID- 12849620 TI - Screening for colorectal cancer using the faecal occult blood test: a critical literature review. AB - Colorectal cancer (CRC) is a major health problem whose outcome is crucially dependent on the stage of disease at the time of diagnosis. Consequently, early detection of CRC by mass population screening is the subject of much international research and debate. A critical literature review was undertaken to determine the effectiveness of faecal occult blood (FOB) testing in CRC screening. Four issues were addressed: an appraisal of available FOB tests, their impact on CRC mortality, the appropriate screening interval and the factors influencing public compliance with FOB screening. A computerized literature search was conducted, supplemented by cross-referencing and hand searching. Forty studies which fulfilled pre-defined inclusion and exclusion criteria were appraised. A thorough assessment of methodology, combined with an applied hierarchy of evidence, resulted in the selection of 22 studies for review. The evidence from this review is that no single (currently used) FOB test provides optimal screening performance, that biennial FOB screening reduces CRC mortality by between 6-18%, that there is a small advantage in annual screening and that better dissemination of information can improve compliance rates. An appreciation of the current evidence and issues surrounding CRC screening is important to nurses in their capacity as health promoters and educators. Definitive answers to many important questions regarding FOB screening for CRC do not currently exist. Before the concept of a national screening programme could be supported, further research is needed, the key elements of which must include the assessment of more sensitive and specific FOB tests and the promotion of population compliance. PMID- 12849621 TI - An evaluation of the quality of a chemotherapy administration service established by nurses in an oncology day care centre. AB - Traditionally, chemotherapy administration has been a medical role in Hong Kong. In 1998, two chemotherapy nurses took over part of this role within a major oncology day care centre after completing a course in chemotherapy administration and patient care. The aim of this study was to measure and evaluate the quality of care by using a self-developed audit form and the Worthing Chemotherapy Satisfaction Questionnaire (WSCQ). All records of patients who had had i.v. cannulas inserted by nurses over a period of one year were audited for the accuracy of the i.v. cannulation (n=2217). Also, patient satisfaction was assessed in 50 patients. The results showed that the accuracy of i.v. cannulation was kept constantly above 80% from the eighth month of auditing onwards. It was also found that patients preferred the use of a butterfly needle for the administration of chemotherapy. The study revealed that the level of information received regarding treatment (r=0.36, P=0.011) and satisfaction of practical advice on caring for side effects (r=0.35, P=0.012) were determinants of patients' overall satisfaction. Furthermore, anxiety when patients received chemotherapy could also be associated with overall satisfaction (r=-0.27), although the P value was of borderline significance only (P=0.58). In conclusion, the study reflected a high level of quality of care in relation to this element of nurse-led practice. PMID- 12849622 TI - Important aspects of care and assistance for parents of children, 0-18 years of age, on or off treatment for cancer. Parent and nurse perceptions. AB - Important aspects of care and assistance for parents of children (0-18 years) diagnosed with cancer were investigated. Parents (N=114) and nurses (N=121) were asked the following questions: 'What caring aspects are important for you/the parent to feel cared for?' and 'What help, if any, do you/the parent need outside the hospital?' Nurses were asked to answer for a certain parent. Data were analyzed by content analysis. The following aspects of care were identified: accessible care, clinical competence, continuity, emotional support, information, participation in care, participation in decision making, physical ambience, shift in parenting roles, social competence, temporary assistance, own child is well cared for, and time. Most parents mentioned the importance of social competence and information, most nurses mentioned the importance of information and emotional support. The following aspects of assistance were identified: accessible care, emotional support, financial support, instrumental support, and support to live a normal family/social life. A third of the parents and a fourth of the nurses were of the opinion that the parent did not need any assistance. Most parents who mentioned a need of assistance mentioned a need for emotional and instrumental support. Most nurses mentioned a parental need for emotional support and accessible care. PMID- 12849623 TI - Cancer-related fatigue in children and teenagers: the unrecognized symptom? PMID- 12849624 TI - An evaluation of the impact of a self-care coping intervention on psychological and clinical outcomes in adolescents with newly diagnosed cancer. AB - The demands of cancer treatment are particularly challenging for newly diagnosed adolescents. If attempts to cope with these demands are unsuccessful, adolescents may not comply with or may refuse treatment. As a result, their chances of survival are decreased. The purpose of this study, guided by the Adolescent Self Sustaining Model, was to determine the effects of a three-part educational intervention designed to facilitate copying on psychological (hopefulness, hopelessness, self-esteem, self-efficacy and symptom distress) and clinical outcomes (treatment toxicity) among adolescents newly diagnosed with cancer. This two-site study used a longitudinal experimental two-group design with adolescents randomly assigned to the intervention or control group. Four measurement points spanning the first 6 months of treatment were included in the design. Of 93 eligible adolescents, 78 (46 females and 32 males) agreed to participate. No statistically significant differences between the intervention and control groups, or between male and female participants, were detected at any measurement point. Differences in scores over time within groups were noted. Explanations for the lack of group differences are offered, as are recommendations for strengthening the intervention and design for future testing. PMID- 12849627 TI - 'I don't have any energy': The experience of fatigue in young adult survivors of childhood cancer. AB - Although it is speculated that fatigue occurs equally in adults, children and adolescents with cancer, little research exists to substantiate this view. Evidence that fatigue continues after treatment is limited in both the adult and paediatric oncology literature. Due to the current lack of knowledge, more information on the phenomenology of fatigue of childhood cancer survivors is desirable. Therefore a study was conducted to explore the concept of fatigue from a survivor's perspective. A semi-structured interview was conducted with a purposeful sample of 35 long-term survivors of childhood cancer who reported feeling extremely fatigued. The topics which were covered during the interview included the nature, onset and pattern of fatigue, sleep rest pattern, what helps with fatigue and what does not help, and the impact of fatigue on their daily life. Most survivors who were diagnosed with cancer in their adolescence identified fatigue as a significant side-effect of the treatment. The majority of survivors who were toddlers or preschooler at the time of cancer treatment mentioned that, as far as they could recall, they had suffered from fatigue their entire life. The course of fatigue during the day differed among the survivors, although the majority reported to be fatigued when waking up in the morning. None of the survivors reported sleep problems. Many survivors slept 9 hours or more. Fatigue was defined by all respondents as having a negative impact on their daily lives. Findings revealed that fatigue is a serious problem for some young adult survivors of childhood cancer and affects many aspects of quality of life. PMID- 12849628 TI - Psychosocial impact of recurrent cancer. AB - The experience of cancer recurrence is multifaceted and is influenced by social support, family support and medical and nursing intervention. The diagnosis of recurrence is often more traumatic than the initial diagnosis and the future for the patient becomes less certain. A chronic illness model provides a useful framework for understanding the experience of recurrent cancer. People who are diagnosed with recurrent cancer recognize that it is a time of re-evaluation, transformation and redefinition. Relationships take on an important significance and informational and support needs are paramount. Much of the literature suggests that the role transformation, or the adaptation of the individual to their circumstances, is a central theme of the experience of cancer recurrence. The development of a specific model to assess the role transformation that patients appear to experience in the face of the uncertainty of their recurrent disease, would be invaluable when developing a theoretical framework to guide future research and nursing care. PMID- 12849629 TI - Cancer patients', family members' and professional helpers' conceptions and beliefs concerning death. AB - The purpose of this study was to examine the attitudes of different groups of people towards death, as well as their opinions and beliefs concerning life after death. The study is part of a larger research project in which cancer patients' suffering and coping were under examination. Data were collected in semi structured focused interviews with 32 patients with incurable cancer, 13 family members, 13 nurses and 13 doctors from two central hospitals and four community health centres. The interviews were tape-recorded and transcribed verbatim. Data were analysed by content analysis. Most of the interviewees were not afraid of death. The professionals were slightly more often afraid of death than the patients and the family members. The participants gave many reasons for their opinions. The interviewees were not very interested in reading death-related literature. Only nurses had read a great deal of literature on death. Most of the participants believed in the existence of God and in life after death. The participants had different kinds of conceptions of the events after death. The doctors differed from the other groups in that they read less literature on death and had less faith in God and in life after death. PMID- 12849630 TI - Taking practice forward in paediatric oncology: the impact of a newly developed education programme for nurses working in shared-care hospitals. AB - Fundamental to any change or development in health care is the quality of the service received by patients and their families. Practitioner-led initiatives that are designed to improve the quality of care are more likely to succeed if they are underpinned with appropriate educational input. A structured programme in paediatric oncology nursing was developed to meet the needs of nurses working in District General Hospitals. Nurses' perceptions of the consequences of undertaking the programme were explored using a focus group approach. Findings have shown that there have been significant changes to nurses' roles and this in turn has had an impact on the individual and the organization. It was perceived that there are positive outcomes for both the service and quality of care for children with cancer and their families. The shared-care nurses believe that this has been the result of improved knowledge, skill and expertise, better communication between care providers, and the collaborative frameworks which have evolved. PMID- 12849631 TI - Sperm banking and fertility concerns: enhancing practice and the support available to men with cancer. PMID- 12849632 TI - Palliative care for people who have a learning disability: exploring the professional needs of qualified district nurses. PMID- 12849633 TI - Finding a role for cancer nurses in research. PMID- 12849634 TI - Perceived symptoms and quality of life in women with breast cancer receiving radiation therapy. AB - The purpose of this study was to describe symptoms, side-effects and quality of life (QoL) of women with breast cancer during and following treatment with radiation therapy. The sample consisted of 134 women with breast cancer. Symptoms were measured using a modified version of the Oncology Treatment Toxicity Tool (OTTAT) and QoL was measured using the Cancer Rehabilitation Evaluation System short form (CARES-sf). The results showed an increase in experienced symptoms and their severity as the treatment progressed. QoL was perceived as poorest at baseline before treatment had started. During the treatment, QoL scores leveled out and an improvement could be seen after completion of treatment. The study findings provide directions and suggestions for assessment and management of perceived symptoms for women receiving radiation therapy for breast cancer from the second week and up to 2 weeks after completion of therapy is the critical time-period for targeting interventions for experienced symptoms and side-effects from radiation therapy. PMID- 12849637 TI - Identifying patient concerns: improving the quality of patient visits to the oncology out-patient department--a pilot audit. AB - Much emphasis is placed on reducing waiting times to improve the quality of patient visits; however, it has been pointed out that improving the quantity of patients seen does not necessarily equate with quality (Hart 1996). This paper describes the design, implementation and evaluation of a pilot feasibility project and audit to improve the quality of patient's follow-up visits to the oncology out-patient department through the use of a 'concerns checklist'. Questionnaires administered to patients and health-care professionals involved in the project were audited. The results indicated that patients found completing the checklist an improvement to the service and helpful with regard to communication, information and staff-patient relationships. PMID- 12849638 TI - Participation of relatives in the care of cancer patients. AB - This study aims to find out how important it is for relatives to take part in the care of cancer patients and to see to what extent they actually do take part. Participation is defined as consisting of concrete care activities, emotional support and participation in decision-making concerning the patient's care. The sample comprised 168 relatives of cancer patients from oncology wards all over Finland. Data were collected with a questionnaire specifically developed for this study. Nonparametric tests were used for statistical analyses. The results showed that the most important aspect of relatives' participation was supporting the patient emotionally; providing concrete care and participation in decision-making were both considered less important. Over 80% of the relatives said they gave a considerable amount of support to the patient. Most relatives said that they visited the patient in hospital, talked to the patient about their concerns and were near them whenever they felt they needed that. On the other hand, relatives did not take part in discussions about care, nor were they informed about such a possibility. PMID- 12849639 TI - Establishing nursing research priorities on a paediatric haematology, oncology, immunology and infectious diseases unit: a Delphi survey. AB - For research to be successfully integrated and applied to practice, ownership and identification must come from those who are most likely to implement research into practice. This was one of the reasons for undertaking a Delphi survey to identify and rank the research priorities for clinical nursing research in a paediatric haematology, oncology, immunology and infectious diseases unit. The 'Nurses' Research Group' initiated the survey as a first step towards developing a strategy for evidence-based nursing. Four members of the research group volunteered to establish a working party to undertake the survey. This paper describes a four-round Delphi survey. The survey questionnaire was sent to all nursing staff on the unit. The initial process identified 151 research topics/themes. Through a process of refinement the priority list was reduced to 89. Repeat rounds were completed, culminating in the identification of four top priority areas of: symptom management, negotiation of care between the child and family, quality-of-life issues and retention of staff. The findings indicate directions for future clinical nursing research that will benefit specialist nurses, children and young people and their families. This paper provides a detailed account of the method, procedure and outcomes of the Delphi survey. The limitations of the Delphi survey method are also addressed and in this survey these included time (the length of time it took to complete the survey and time needed to complete each questionnaire), maintaining motivation of respondents, and the influence of researchers working in the research setting. PMID- 12849640 TI - Handling bad news: an innovative training approach. PMID- 12849642 TI - Health promotion and cancer nursing. PMID- 12849641 TI - A longitudinal study of cancer-related fatigue in patients with breast cancer: research in progress. PMID- 12849643 TI - A preliminary study of patients' perceptions of routine follow-up after treatment for breast cancer. AB - Early detection and longer survival have led to increasing numbers of patients requiring follow-up after treatment for breast cancer, yet there is little consensus as to how this should be achieved. Breast cancer patients have needs that change over time but the current system of follow-up is traditionally routinized and lacks an individualized approach. This preliminary study was designed to ascertain patients' perceptions of routine follow-up care after completion of treatment for breast cancer. A cross-sectional survey of a stratified systematic sample of patients was utilized. Data were collected using semi-structured, taped interviews. The tapes were inductively analysed and coded to ascertain predominant themes. Twenty-four patients were recruited. Analysis indicated that follow-up examinations were hurried (18 patients), investigations were not reassuring (11) and that the lack of continuity was unacceptably poor (22). Many patients (19) felt uncomfortable expressing emotional concerns or asking questions. The majority (18) stated that they would prefer to receive all or part of their follow-up from a breast care nurse. These findings have implications for service provision in terms of quality and cost-effectiveness. On the basis of these results a patient-focused, nurse-led intervention is being evaluated against traditional medical follow-up for patients with breast cancer. PMID- 12849645 TI - A European cancer nursing curriculum for a post-basic course in cancer prevention: outcomes of a collaborative project. AB - The core curriculum distance learning project in cancer prevention was funded by Europe Against Cancer and the University of Southampton and supported by the European Oncology Nursing Society. The commitments of the project were to collaborative working, involvement of as many EU countries as possible, joint writing and design of the materials, and a flexible and useful end-product. The ambitions of the project were realized by the collective production of materials on cancer prevention that are available either as a teaching resource in English, French or Portuguese, or an accredited post-graduate course by distance learning in English. This paper will focus on the tensions and issues arising out of the collaborative process; these were connected to funding, translation, dissemination and evaluation. PMID- 12849646 TI - Clinical research in paediatric oncology and the role of the research nurse in the UK. AB - Survival rates for childhood cancer have improved considerably as new drugs, treatment protocols and supportive therapies have developed through clinical trials. Such studies take a considerable amount of time and organization and alongside scientific and medical staff the research nurse plays a vital role. The facets of this role will be outlined in this paper. Involvement begins at an early stage of study development and includes applying for Research Ethics Committee approval of the project. Once the study has opened the research nurse is then responsible for patient recruitment, monitoring and follow-up. The research nurse works within a team but also with a degree of autonomy ensuring that standards of patient care are maintained by adhering to guidelines for clinical research in general and those aimed at children specifically. Providing detailed information and support to the child and family, staff and outside agencies are other notable aspects of the post. The role of the research nurse continues to develop, as clinical trials maintain a significant role in improving the treatment for childhood cancer. PMID- 12849647 TI - Attitudes to cancer of medical and surgical nurses in a district general hospital. AB - The personal beliefs and attitudes of nurses can have serious implications for their practice. Yet there are a lack of studies on nurses' attitudes to cancer in the UK in general, and in Northern Ireland specifically. The need for such studies is further emphasized by the importance of nurses' role in the Cancer Units of district general hospitals, as proposed by the Calman-Hine Report (Department of Health 1995). The aim of this study was to assess the attitudes to cancer of medical and surgical nurses who care for cancer patients in one district general hospital in Northern Ireland. A questionnaire comprising the Cancer Attitude Scale (Corner 1993) was administered to 106 qualified nurses. The response rate was 68.9% (n=73). Responses to items on the Cancer Attitude Scale show that while generally these nurses held positive attitudes, a high proportion also held negative views about cancer. Of particular concern were their attitudes to the value of active treatments. The implications of these findings are discussed. PMID- 12849648 TI - Problems of infertility following cancer therapy in childhood and adolescence. AB - Hormones regulate ovarian and testicular function. These are produced by the anterior lobe of the pituitary gland, the hypothalamus and by the sex organs. Impaired gonadal function may result from damage to any of these structures and the toxic effects of chemotherapy and radiotherapy can cause this damage. This paper will discuss the effects these treatments can have on adults who have survived a childhood cancer, and describe the subsequent implications for clinical practice. PMID- 12849649 TI - Randomized trials in the treatment of cancer. PMID- 12849650 TI - Support for patients with breast cancer: evaluation of needs. PMID- 12849651 TI - Glycoprotein IIb/IIIa inhibition in the setting of acute ST-segment elevation myocardial infarction. AB - Glycoprotein (GP) IIb/IIIa inhibitors have been extensively studied in the setting of percutaneous coronary intervention (PCI) and in the management of non ST-segment elevation acute coronary syndromes. However, the use of GP IIb/IIIa inhibitors is less well established in the setting of acute ST-segment elevation myocardial infarction (MI). Multiple nonrandomized studies suggest that combination therapy with GP IIb/IIIa inhibitors and thrombolytic agents leads to increased rates of TIMI 3 flow. However, two clinical trials involving over 22,000 patients demonstrated that combination therapy is associated with only modest reductions in major adverse cardiac events, does not reduce mortality, and is associated with an increase in bleeding. In the setting of primary PCI, four clinical trials involving over 3,000 patients demonstrated that GP IIb/IIIa inhibition results in a significant decrease in the need for urgent target vessel revascularization but not in reductions of death or recurrent MI. Thus, GP IIb/IIIa inhibition may provide only limited benefits in the setting of acute ST segment elevation MI. PMID- 12849652 TI - Early and long-term clinical outcomes associated with reinfarction following fibrinolytic administration in the Thrombolysis in Myocardial Infarction trials. AB - OBJECTIVES: We hypothesized that early recurrent myocardial infarction (MI) following fibrinolytic administration would be assessed with higher mortality at both 30 days and 2 years. BACKGROUND: Although early recurrent MI after fibrinolytic therapy has been associated with increased early mortality in the acute MI setting, its relation to long-term mortality has not been fully explored. METHODS: Mortality data were ascertained in 20,101 patients enrolled in the Thrombolysis In Myocardial Infarction (TIMI) 4, 9, and 10B and Intravenous NPA for the Treatment of Infarcting Myocardium Early (InTIME-II) acute MI trials. RESULTS: The frequency of symptomatic recurrent MI during the index hospitalization was 4.2% (836/20,101). Recurrent MI during the index hospital period was associated with increased 30-day mortality (16.4% [137/836] vs. 6.2% [1,188/19,260], p < 0.001). Likewise, recurrent MI was associated with a sustained increase in mortality up to two years, even after adjustments were made for covariates known to be associated with mortality and recurrent MI (hazard ratio 2.11, p < 0.001). However, this higher mortality at 2 years was due to an early divergence in mortality by 30 days and was not due to a significant increase in late mortality between 30 days and 2 years (4.38% [31/707] vs. 3.76% [685/18,206], p = NS). Percutaneous coronary intervention during the index hospitalization was associated with a lower rate of in-hospital recurrent MI (1.6% vs. 4.5%, p < 0.001) and lower two-year mortality (5.6% vs. 11.6%, p < 0.001). Performance of coronary artery bypass graft surgery was also associated with a lower recurrent rate of MI (0.7% vs. 4.3%, p < 0.001) and lower two-year mortality rate (7.95% vs. 10.6%, p = 0.0008). CONCLUSIONS: Early recurrent MI is associated with increased mortality up to two years. However, most deaths occur early, and the risk of additional deaths between the index hospital period and two years was not significantly increased among patients with recurrent MI. Percutaneous coronary intervention during the index hospitalization was associated with a lower risk of recurrent MI and a lower risk of two-year mortality. PMID- 12849653 TI - "Watchful waiting" after thrombolysis: it's time for a re-evaluation. PMID- 12849654 TI - Maintenance of sinus rhythm in patients with atrial fibrillation: an AFFIRM substudy of the first antiarrhythmic drug. AB - OBJECTIVES: This study evaluated the efficacy of antiarrhythmic drugs for the treatment of atrial fibrillation (AF). BACKGROUND: The most effective and safest antiarrhythmic drug for the treatment of AF is unknown. METHODS: The Atrial Fibrillation Follow-up Investigation of Rhythm Management (AFFIRM) study compared two treatment strategies--rate control and rhythm control--in patients with AF and risk factors for stroke or death. This substudy, performed in patients randomized to rhythm control, compared different antiarrhythmic drugs by randomly assigning the first drug treatment to: 1) amiodarone, 2) sotalol, or 3) a class I drug. The primary end point was the proportion of patients alive, in sinus rhythm, with no additional cardioversions and still taking the assigned drug at one year. Comparisons were made between patients eligible for each of three drug pairs. RESULTS: At one year, in 222 patients randomized between amiodarone and class I agents, 62% were successfully treated with amiodarone, compared with 23% taking class I agents (p < 0.001). In 256 patients randomized between amiodarone and sotalol, 60% versus 38% were successfully treated, respectively (p = 0.002). In 183 patients randomized between sotalol and class I agents, 34% versus 23% were successfully treated, respectively (p = 0.488), although this portion of the substudy was stopped early when amiodarone was shown to be better than class I agents. Sinus rhythm was achieved in nearly 80% of patients at one-year follow-up with serial therapy. Adverse effects were common. CONCLUSIONS: Amiodarone was more effective at one year than either sotalol or class I agents for the strategy of maintenance of sinus rhythm without cardioversion. PMID- 12849655 TI - Restoring sinus rhythm in atrial fibrillation: a Pyrrhic victory? PMID- 12849656 TI - Long-term outcome of patients with silent versus symptomatic ischemia six months after percutaneous coronary intervention and stenting. AB - OBJECTIVES: We sought to evaluate the incidence of silent ischemia versus symptomatic ischemia six months after percutaneous coronary intervention (PCI) and its impact on prognosis and to test the utility of myocardial perfusion single-photon emission computed tomography (SPECT), or MPS, for risk stratification in these patients. BACKGROUND: Silent ischemia is frequent after PCI. However, little is known about silent ischemia and long-term outcome after PCI and stenting. METHODS: In 356 consecutive patients with successful PCI and stenting and follow-up MPS after six months, long-term follow-up (4.1 +/- 0.3 years) was performed. The MPS images were interpreted by defining summed stress, rest, and difference scores (summed difference score [SDS] = extent of ischemia) and related to symptoms and outcome. Critical events included cardiac death, myocardial infarction, and target vessel revascularization. RESULTS: Eighty-one patients (23%) had evidence of target vessel ischemia, which was silent in 62%. The only independent predictor of silent ischemia was SDS (odds ratio 0.64, p = 0.001). During follow-up, 67 critical events occurred. For patients with an SDS of 0, 1-4, and >4, the critical event rates were 17%, 29%, and 69%, respectively. Similarly, patients without ischemia, silent ischemia, and symptomatic ischemia had 17%, 32%, and 52% of critical events, respectively. Diabetes (relative risk 1.98, p = 0.03) and SDS (relative risk 1.2, p < 0.001) were independent predictors of critical events. The MPS image added incremental information for the prediction of critical events. CONCLUSIONS: Six months after PCI and stenting, 23% of patients had target vessel ischemia, which was silent in 62%. Silent ischemia predicted a worse outcome than did no ischemia and tended to have a better outcome than symptomatic ischemia. This was closely related to the extent of ischemia. The SDS added incremental value to pre-scan findings with respect to diagnosis and prognosis, indicating the utility of MPS for risk stratification after PCI and stenting. PMID- 12849657 TI - Silent ischemia: unsafe at any time. PMID- 12849658 TI - Early use of glycoprotein IIb/IIIa inhibitors in non-ST-elevation acute myocardial infarction: observations from the National Registry of Myocardial Infarction 4. AB - OBJECTIVES: We sought to identify patient and hospital features associated with early glycoprotein (GP) IIb/IIIa inhibitor therapy for non-ST-elevation (NSTE) myocardial infarction (MI) and to relate this treatment to in-hospital outcomes. BACKGROUND: Glycoprotein IIb/IIIa inhibitors have improved outcomes in randomized trials of NSTE MI, leading national treatment guidelines to recommend their use. Their actual use, safety, and effectiveness have not been well characterized beyond trial populations. METHODS: We studied 60,770 patients with NSTE MI treated between July 2000 and July 2001 at 1,189 hospitals in a U.S. registry. Using logistic regression, we identified patient and hospital features associated with GP IIb/IIIa inhibition within 24 h after presentation. We also compared outcomes by early treatment versus no treatment after adjusting for patient and hospital characteristics and treatment propensity. RESULTS: Only 25% of eligible patients received early GP IIb/IIIa therapy. Elderly patients, women, minority patients, and those without private insurance received such therapy less often than their counterparts. Treated patients had lower unadjusted in-hospital mortality (3.3% vs. 9.6%, p < 0.0001) remaining significantly lower after adjustment for patient risk, treatment propensity, and hospital characteristics (adjusted odds ratio, 0.88; 95% confidence interval, 0.79 to 0.97). Hospitals that adopted early GP IIb/IIIa inhibition more rapidly also had lower adjusted mortality rates than those slower to adopt such therapy. CONCLUSIONS: Glycoprotein IIb/IIIa inhibitor therapy appears to be underused in early management of NSTE MI patients. Because this therapy is associated with better outcomes, it represents a target for quality improvement. PMID- 12849659 TI - Early glycoprotein IIb/IIIa inhibitors in patients with non-ST-segment elevation myocardial infarction: are we ready to open the floodgates? PMID- 12849660 TI - Do associations with C-reactive protein and extent of coronary artery disease account for the increased cardiovascular risk of renal insufficiency? AB - OBJECTIVES: We sought to determine whether the association of higher C-reactive protein levels (CRP) and more extensive coronary artery disease (CAD) explains the high cardiovascular risk of renal insufficiency (RI). BACKGROUND: Renal insufficiency and renal failure (RF) have been associated with increased cardiovascular risk in several studies, and it has been suggested that this association may be due to higher CRP levels and greater extent of CAD. To what extent CRP or severity of CAD explains this risk is uncertain. METHODS: A total of 1,484 patients without myocardial infarction (MI) undergoing angiography were entered and followed for 3.0 +/- 1.6 years; RI and RF were defined as estimated glomerular filtration rates (GFR) of 30 to 60 and <30 ml/min; CRP was measured by immunoassay and > or = 1.0 mg/dl defined as elevated. A CAD score was determined by extent and severity of angiographic disease. Multivariate Cox regressions were performed using seven standard risk factors, homocysteine, GFR, CRP, and CAD score. RESULTS: Mean age was 64 years, and 67% were men; CAD was absent in 24%, mild in 11%, and severe (> or =70% stenosis) in 60%; CRP and CAD scores increased with declining renal function (median CRP: 1.2, 1.4, 2.2 mg/dl, p < 0.001 and CAD score: 8.1, 8.7, 9.3, p = 0.008 for no-RI, RI, and RF). During follow-up, 208 patients (15%) died or had nonfatal MI. Unadjusted hazard ratio (HR) for death/MI was 2.3 for RI and 5.1 for RF (p < 0.0001). Adjustment for CRP (HR, 2.2, 4.5), CAD score (HR, 2.1, 5.1), and all other risk factors (HR, 1.7, 4.5) had minimal or modest impact on RI and RF risk; HR increased to 5.4 (p < 0.001) for presence of both elevated CRP and RI/RF. CONCLUSIONS: Renal insufficiency, CRP, and angiographic CAD, although correlated, are largely independent predictors of cardiovascular risk, suggesting the importance of both inflammation and as yet undefined RI-related risk factors. PMID- 12849661 TI - Incremental value of combined perfusion and function over perfusion alone by gated SPECT myocardial perfusion imaging for detection of severe three-vessel coronary artery disease. AB - OBJECTIVES: We hypothesized that combining functional assessment to perfusion enhances the ability of electrocardiographic gating Tc-99m sestamibi single photon emission computed tomography (gated SPECT) myocardial perfusion imaging (MPI) to detect defects in multiple vascular territories in patients with severe three-vessel coronary artery disease (3VD). BACKGROUND: In patients with 3VD, perfusion defects in multiple vascular territories may not always be evident due to globally reduced perfusion. METHODS: Gated SPECT MPIs were interpreted sequentially with perfusion first, followed by combined perfusion/function, in 143 patients with angiographic 3VD and a control group of 112 non-3VD patients. All patients underwent coronary arteriography within one month of MPI. RESULTS: In 3VD patients, combined perfusion/function analysis yielded significantly greater numbers of abnormal segments/patient (6.2 +/- 4.7 vs. 4.1 +/- 2.8, p < 0.001) and more defects in multiple vascular territories (60% vs. 46%, p < 0.05) than perfusion alone. In the control group, there were no differences between the combined perfusion/function and perfusion alone interpretations. Multivariate analysis of 15 different clinical, stress, and scintigraphic variables in all patients revealed age (p < 0.0001) and number of abnormal vascular territories by combined perfusion/function (p < 0.0001) to be the most powerful predictors of 3VD. Addition of functional data to clinical, stress, and perfusion yielded a significant increase in the predictive value of 3VD (global chi-square: 131.7 vs. 89.8, p < 0.00001). Specificity of combined perfusion/function analysis was not lower than perfusion alone (72% vs. 69%, p = NS). CONCLUSIONS; Adjunctive assessment of function with perfusion by gated SPECT MPI enhances the detection of defects in multiple vascular territories in patients with severe 3VD, without adversely affecting its specificity. PMID- 12849662 TI - Statins enhance postischemic hyperemia in the skin circulation of hypercholesterolemic patients: a monitoring test of endothelial dysfunction for clinical practice? AB - OBJECTIVES: The present study aims to investigate whether laser Doppler flowmetry can be used to monitor improvements in vascular function during statin therapy. BACKGROUND: Endothelial dysfunction is an early feature of atherosclerosis in hypercholesterolemic patients and can be improved by statins. There are several methods to assess endothelial function in vivo, none of them being feasible in everyday practice. METHODS: Skin perfusion, measured by laser Doppler flowmetry, was assessed at rest and during reactive hyperemia. Nineteen hypercholesterolemic patients (age 42 to 73 years, total cholesterol 5.4 to 9.6 mmol/l) were studied before and during statin therapy. To further investigate the mechanisms, postischemic skin hyperemia was measured before and after intradermal injection of the nitric oxide synthase inhibitor L-NAME and its inactive isoform D-NAME (0.5 micromol/10 microl each). On a separate day, the healthy volunteers were reexamined before and 2 h after 1,000 mg aspirin. RESULTS: Postischemic skin blood flow was markedly reduced in hypercholesterolemic patients (45 +/- 11%) compared with healthy controls (238 +/- 20%, p < 0.0001) and improved after statin therapy (113 +/- 15%, p = 0.0005 vs. pre-treatment). In the healthy volunteers, the hyperemic responses were not significantly different after L-NAME and D-NAME. Aspirin reduced hyperemia from 274 +/- 49% to 197 +/- 40% (p = 0.025). CONCLUSIONS: Reactive hyperemia of the skin microcirculation can be easily and reproducibly assessed by laser Doppler flowmetry. Vasodilator prostaglandins are the major mediators of postischemic skin hyperemia, which is impaired in hypercholesterolemic patients and can be enhanced by cholesterol lowering therapy. Thus, laser Doppler flowmetry may represent a tool to assess and monitor vascular function during therapy in everyday practice. PMID- 12849663 TI - Statins, skin, and the search for a test of endothelial function. PMID- 12849664 TI - Are lipid-lowering drugs also antiarrhythmic drugs? An analysis of the Antiarrhythmics versus Implantable Defibrillators (AVID) trial. AB - OBJECTIVES: This study sought to evaluate the antiarrhythmic effects of lipid lowering drug therapy as assessed by ventricular tachyarrhythmia (ventricular tachycardia [VT]/ventricular fibrillation [VF]) recurrences recorded by an implantable cardioverter defibrillator (ICD) in patients with atherosclerotic heart disease (ASHD). BACKGROUND: Randomized trials of lipid-lowering drugs suggest reduction of sudden death (SD) in patients with ASHD. Because SD is usually secondary to VT/VF, this observation suggests that lipid-lowering therapy has antiarrhythmic effects. METHODS: The probability of VT/VF recurrence in patients with ASHD treated with an ICD in the Antiarrhythmics Versus Implantable Defibrillators (AVID) trial who did not receive lipid-lowering drug therapy (n = 279) was compared with that in patients who received early and consistent lipid lowering therapy (n = 83). In addition, all-cause mortality and cardiac mortality of all patients in the AVID trial with ASHD who did not receive lipid-lowering therapy (n = 564) were compared with that of those who received early and consistent lipid-lowering therapy (n = 149). RESULTS: Using multivariate analyses, lipid-lowering therapy was associated with a reduction in the relative hazard for VT/VF recurrence of 0.40 (95% confidence interval [CI] 0.15 to 0.58) (adjusted p = 0.003) in the ICD subgroup. Lipid-lowering therapy was also associated with a reduction in the relative hazard for all-cause mortality of 0.36 (95% CI 0.15 to 0.68) (adjusted p = 0.03) and a reduction in the relative hazard for cardiac mortality of 0.39 (95% CI 0.16 to 0.78) (adjusted p = 0.04) in the larger study population. CONCLUSIONS: In patients with ASHD who have received an ICD, lipid-lowering therapy is associated with reduction in the probability of VT/VF recurrence, suggesting that part of the benefit of lipid-lowering therapy may be due to an antiarrhythmic effect. PMID- 12849665 TI - Lipid-lowering therapy for prevention of ventricular tachyarrhythmias. PMID- 12849666 TI - The prevalence of atrial fibrillation in incident stroke cases and matched population controls in Rochester, Minnesota: changes over three decades. AB - OBJECTIVES: We sought evidence of a change in the prevalence of atrial fibrillation (AF) over a 30-year period among residents of Rochester, Minnesota. BACKGROUND: Atrial fibrillation is increasingly encountered in clinical practice, but there is limited data on secular trends of AF over time. METHODS: Within a longitudinal case-control study of ischemic stroke, the prevalence of AF and of selected comorbid conditions among incident stroke cases and age- and gender matched controls between 1960 and 1989 was determined. RESULTS: The mean age +/- standard deviation for the 1,871 stroke cases (45% men) and matched controls was 75 +/- 11 years. For cases, age-adjusted estimates of AF prevalence for 1960 to 1969, 1970 to 1979, and 1980 to 1989 were 11%, 13%, and 16%, respectively, for men, and 13%, 16%, and 20% for women. For controls, the rates were 5%, 8%, and 12%, respectively, for men, and 4%, 6%, and 8% for women. Increasing AF prevalence was associated with increasing age (doubling of odds per decade of age in both cases and controls) and calendar time adjusted for age and gender (cases: odds ratio [OR] per 5 years 1.13, 95% confidence interval [CI], 1.05 to 1.22; controls: OR per 5 years 1.24, 95% CI 1.12 to 1.37). The rates of increase with calendar time were significant for cases (p = 0.001) and controls (p < 0.001) and comparable between the genders. CONCLUSIONS: The prevalence of AF increased significantly in ischemic stroke patients and their controls from 1960 to 1989 in Rochester, Minnesota, independent of age and gender. The rate of increase did not differ significantly between men and women. PMID- 12849667 TI - Atrial fibrillation--more evidence that it's here to stay. PMID- 12849668 TI - Modulating effects of age and gender on the clinical course of long QT syndrome by genotype. AB - OBJECTIVES: We aimed to determine whether long QT syndrome (LQTS) genotype has a differential effect on clinical course of disease in male and female children and adults after adjustment for QTc duration. BACKGROUND: Genotype influences clinical course of the LQTS; however, data on the effect of age and gender on this association are limited. METHODS: The LQTS genotype, QTc duration, and follow-up were determined in 243 cases of LQTS caused by the KCNQ1 potassium channel gene mutations (LQT1), 209 cases of LQTS caused by the HERG potassium channel gene mutations (LQT2), and 81 cases of LQTS caused by the SCN5A sodium channel gene mutation (LQT3) gene carriers. The probability of cardiac events (syncope, aborted cardiac arrest, or sudden death) was analyzed by genotype, gender, and age (children < or = 15 years and adults 16 to 40 years). In addition, the risk of sudden death and lethality of cardiac events were evaluated in 1,075 LQT1, 976 LQT2, and 324 LQT3 family members from families with known genotype. RESULTS: During childhood, the risk of cardiac events was significantly higher in LQT1 males than in LQT1 females (hazard ratio [HR] = 1.72), whereas there was no significant gender-related difference in the risk of cardiac events among LQT2 and LQT3 carriers. During adulthood, LQT2 females (HR = 3.71) and LQT1 females (HR = 3.35) had a significantly higher risk of cardiac events than respective males. The lethality of cardiac events was highest in LQT3 males and females (19% and 18%), and higher in LQT1 and LQT2 males (5% and 6%) than in LQT1 and LQT2 females (2% for both). CONCLUSIONS; Age and gender have different, genotype-specific modulating effects on the probability of cardiac events and electrocardiographic presentation in LQT1 and LQT2 patients. PMID- 12849669 TI - Recognition of far-field electrograms during entrainment mapping of ventricular tachycardia. AB - OBJECTIVES: The goal of this study was to assess entrainment for distinguishing far-field potentials (FFP) due to depolarization of tissue at a distance from the mapping catheter from the local potential (LP) due to depolarization of tissue at the catheter electrode during mapping of ventricular tachycardia (VT). BACKGROUND: Electrograms with multiple peaks commonly complicate mapping and identification of catheter ablation targets in infarcts. METHODS: Retrospective analysis of catheter mapping data from eight patients with prior infarction was performed to evaluate multipotential electrograms at sites where pacing entrained VT. Potentials that were visible and not altered during pacing were defined as FFP. Potentials obscured by the pacing stimulus were designated possible LPs. The criteria for FFP were then assessed in a second cohort of five patients. RESULTS: At 32 of 39 (82%) sites with multiple potentials, entrainment identified one of the potentials as an FFP. Radiofrequency ablation, assessed at 15 sites, reduced the amplitude of LPs by 62%, without significant effect on FFP amplitude. At 56% of sites with multiple potentials, measuring the postpacing interval to an FFP would lead to erroneous classification of the site location relative to the reentry circuit. In prospective evaluation, double potentials were identified at 77 sites in infarcts; entrainment demonstrated an FFP at 66 (86%) sites. CONCLUSIONS: Far-field potentials are common during mapping in infarcts. Many can be distinguished from local potentials by entrainment, improving the accuracy of mapping. PMID- 12849670 TI - Combining low-intensity and maximal exercise test results improves prognostic prediction in chronic heart failure. AB - OBJECTIVES: This study investigated the combination of maximal and low-intensity exercise testing in predicting prognosis in chronic heart failure (CHF), using one single exercise test (two-step protocol). BACKGROUND: Risk assessment based on any single factor has limited accuracy and reproducibility. METHODS: Treadmill exercise testing was performed in 202 consecutive CHF patients (174 male; mean age 52 +/- 11 years) using "breath-by-breath" gas exchange monitoring. Oxygen uptake (VO(2)) kinetics were defined as oxygen deficit (DeltaVO(2) x time [rest to steady state] - Sigma VO(2) [rest to steady state]) and mean response time (MRT = oxygen-deficit/DeltaVO(2)). Peak VO(2) (VO(2)max) was defined as the highest VO(2). Mean follow-up was 873 +/- 628 days. The primary end point was cardiac mortality and the need for urgent heart transplantation. RESULTS: Forty four patients (22%) died and 15 (7%) were urgently transplanted. In both univariate and multivariate analyses, MRT >50 s was the most powerful predictor of the primary end point (hazard ratio [HR] 4.44), followed by predicted VO(2)max <50% (HR 3.50) and resting systolic blood pressure <105 mm Hg (HR 2.49, all p < 0.001). A majority (n = 130 [64%]) had one or none of these risk factors, with a one-year event rate of only 3%. Patients with two risk factors (n = 45 [22%]) were at medium risk (one-year event rate of 33%). Twenty-seven patients (13%) had all three risk factors, with a one-year event rate of 59%. The area under the curve, using the number of risk factors, was 0.86 +/- 0.04 for the primary end point at one year. These results were independent of medication, in particular, beta-blockade. CONCLUSIONS: A combination of low-intensity and maximal exercise test results improves assessment of prognosis in patients with CHF. PMID- 12849671 TI - Exercise testing in heart failure: maximal, submaximal, or both? PMID- 12849672 TI - Preserved response of mitochondrial function to short-term endurance training in skeletal muscle of heart transplant recipients. AB - OBJECTIVES: We sought to determine whether intrinsic mitochondrial function and regulation were altered in heart transplant recipients (HTRs) and to investigate the response of mitochondrial function to six-week endurance training in these patients. BACKGROUND: Despite the normalization of central oxygen transport during exercise, HTRs are still characterized by limited exercise capacity, which is thought to result from skeletal muscle metabolic abnormalities. METHODS: Twenty HTRS agreed to have vastus lateralis biopsies and exercise testing: before and after training for 12 of them and before and after the same control period for eight subjects unwilling to train. Mitochondrial respiration was evaluated on saponin-permeabilized muscle fibers in the absence or presence (maximum respiration rate [V(max)]) of saturating adenosine diphosphate. RESULTS: Mitochondrial function was preserved at the level of sedentary subjects in untrained HTRs, although they showed 28 +/- 5% functional aerobic impairment (FAI). After training, V(max), citrate synthase, cytochrome c oxidase, and mitochondrial creatine kinase (CK) activities were significantly increased by 48%, 40%, 67%, and 53%, respectively (p < 0.05), whereas FAI decreased to 12 +/- 5% (p < 0.01). The control of mitochondrial respiration by creatine and mitochondrial CK was also improved (p < 0.01), suggesting that phosphocreatine synthesis and transfer by the mitochondrial CK become coupled to oxidative phosphorylation, as shown in trained, healthy subjects. CONCLUSIONS: In HTRs, the mitochondrial properties of skeletal muscle were preserved and responded well to training, reaching values of physically active, healthy subjects. This suggests that, in HTRs, immunosuppressive drugs do not alter the intrinsic muscle oxidative capacities and that the patients' physical handicap results from nonmitochondrial mechanisms. PMID- 12849673 TI - Thromboxane inhibition improves renal perfusion and excretory function in severe congestive heart failure. AB - OBJECTIVES: The aim of this study was to evaluate whether thromboxane inhibition can favorably affect renal perfusion and clinical conditions in patients affected by severe heart failure. BACKGROUND: The renal formation of the vasoconstrictor thromboxane A(2) (TxA(2)) is increased during cardiac failure. METHODS: By oral administration of picotamide (a renal TxA(2) synthase and TxA(2)/prostaglandin H(2) receptor inhibitor), we blocked renal TxA(2). Fourteen patients in New York Heart Association functional class IV were studied according to a randomized, double-blinded, cross-over design. Each of the two eight-day periods of testing was preceded by a three-day period during which certain vasoactive medications were stopped. RESULTS: Daily 24-h total urinary thromboxane B(2) (TxB(2)), the stable metabolite of TxA(2), dropped at the end of picotamide treatment (p < 0.01 vs. baseline). Compared with placebo, effective renal plasma flow and the glomerular filtration rate increased (p < 0.01 and p < 0.05, respectively), thus leading to a significant decrease in the filtration fraction (p < 0.01). Renal vascular resistance decreased consistently (p < 0.01). In all patients, picotamide treatment was associated with an increase in diuresis and natriuresis (p < 0.001 vs. baseline). Plasma creatinine decreased (p < 0.05 vs. baseline). Patients also showed improvement in several clinical parameters, including a significant decrease in both pulmonary and venous pressure (p < 0.01 vs. baseline). CONCLUSIONS: These results indicate that renal thromboxane formation plays an important role in renal vascular resistance in patients with severe heart failure, such as those described in the present study. Inhibition of TxA(2) improves renal hemodynamics and kidney function and favorably affects indexes of cardiac performance. PMID- 12849674 TI - Hemodynamic and clinical effects of tezosentan, an intravenous dual endothelin receptor antagonist, in patients hospitalized for acute decompensated heart failure. AB - OBJECTIVES: We sought to investigate the efficacy and safety of tezosentan, a dual endothelin receptor antagonist, in patients hospitalized for acute heart failure (HF). BACKGROUND: Tezosentan has been previously shown to improve hemodynamics in patients with stable chronic HF. METHODS: In a double-blind fashion, 292 patients (cardiac index < or =2.5 l/min per m(2) and pulmonary capillary wedge pressure (PCWP) > or =15 mm Hg) who were admitted to the hospital and in need of intravenous treatment for acute HF and central hemodynamic monitoring were randomized to 24-h intravenous treatment with tezosentan (50 or 100 mg/h) or placebo. Central hemodynamic variables, the dyspnea score, and safety variables were measured. RESULTS: After 6 h of treatment, significantly greater increases in the cardiac index and decreases in PCWP were observed with both tezosentan dosages than with placebo (mean treatment effects at 0.38 and 0.37 l/min per m(2) with 50 and 100 mg/h and -3.9 mm Hg for each dose, respectively; p < 0.0001). This effect was maintained during the remaining infusion and for > or =6 h after treatment cessation. A tendency for an improved dyspnea score and a decreased risk of clinical worsening was observed after 24 h of treatment with each tezosentan dose. Adverse events, more frequent with tezosentan than with placebo (headache, asymptomatic hypotension, early worsening of renal function, nausea, vomiting), were dose-related. CONCLUSIONS: Intravenous tezosentan rapidly and effectively improved hemodynamics in these patients. The similar beneficial effects of the two dosages and the increased dose-related adverse events with the higher dosage suggest that the optimal dosing regimen is <50 mg/h. PMID- 12849675 TI - Prospective echocardiographic diagnosis and surgical repair of anomalous origin of a coronary artery from the opposite sinus with an interarterial course. AB - OBJECTIVES: In this study, we sought to describe the mode of presentation, anatomic features, diagnostic techniques, and surgical outcome in a group of patients with anomalous origin of a coronary artery from the opposite sinus with an interarterial course between the great arteries (AOCA). BACKGROUND: Anomalous origin of a coronary artery from the opposite sinus with an interarterial course is associated with myocardial ischemia and sudden cardiac death, particularly in adolescents and young adults. METHODS: The cardiology database at Children's Hospital of Wisconsin was reviewed to identify all patients diagnosed with AOCA. RESULTS: From September 1997 to August 2002, 10 patients were identified with AOCA; all were children/adolescents (age range, 3 months to 20 years; weight range, 4.7 to 72 kg), and nine were diagnosed prospectively by transthoracic echocardiography (TTE). Symptoms of cardiac ischemia initiated investigation in 4/10 patients at a mean age of 16 +/- 2.8 years; the other six had TTE for suspected congenital heart disease/musculoskeletal chest pain. The left coronary artery originated from the right sinus in six patients, and the right coronary artery originated from the left sinus in four patients. An intramural course of the AOCA within the anterior aortic wall was found in 9/10 patients and was reliably identified by TTE; the other patient had an intramyocardial course of the anomalous coronary. Surgical repair was performed in 8/10 patients at a mean age of 13 +/- 4.7 years. Unroofing of the intramural portion of the AOCA to relocate the ostia in the appropriate sinus was successfully performed in seven patients. All patients status post unroofing were asymptomatic with patent coronary flow by Doppler and normal exercise treadmill testing at a median follow up interval of 1.5 years. CONCLUSIONS: Anomalous origin of a coronary artery from the opposite sinus with an interarterial course is frequently characterized by an intramural course, which can be prospectively identified by TTE. Unroofing the intramural segment without bypass grafting can reliably repair the intramural form of AOCA. PMID- 12849676 TI - What is the most effective method of detecting anomalous coronary origin in symptomatic patients? PMID- 12849677 TI - Oral sildenafil as long-term adjunct therapy to inhaled iloprost in severe pulmonary arterial hypertension. AB - OBJECTIVES: We sought to investigate the impact of adjunct sildenafil on exercise capacity and hemodynamic parameters in patients with pulmonary arterial hypertension (PAH) who fulfilled predefined criteria of deterioration despite ongoing treatment with inhaled iloprost. BACKGROUND: Inhaled iloprost is an effective therapy in PAH. The phosphodiesterase-5 inhibitor sildenafil exerts pulmonary vasodilation and may amplify prostanoid efficacy. METHODS: Of 73 PAH patients receiving long-term inhaled iloprost treatment, 14 fulfilled criteria of deterioration unresponsive to conventional treatment. These patients received adjunct oral sildenafil over a period of nine to 12 months, leaving the inhalative iloprost regimen unchanged. RESULTS: Before iloprost therapy, the baseline 6-min walking distance was 217 +/- 31 m (mean +/- SEM), with an improvement to 305 +/- 28 m within the first three months of iloprost treatment and a subsequent decline to 256 +/- 30 m after 18 +/- 4 months. Adjunct therapy with sildenafil reversed the deterioration and increased the 6-min walk distance to 346 +/- 26 m (p = 0.002, Wilcoxon test) at three months of combined therapy, with a sustained efficacy up to 12 months (349 +/- 32 m, p = 0.002). The distribution of New York Heart Association functional classes (IV/III/II) improved from September 9, 2000, before sildenafil, to January 8, 2003, after nine to 12 months with sildenafil. All hemodynamic variables changed favorably: pulmonary vascular resistance decreased from 2,494 +/- 256 before sildenafil to 1,950 +/- 128 dynes.s.cm(-5).m(2) after three months of adjunct sildenafil (p = 0.036). Two patients died of severe pneumonia during the period of combined therapy. No further serious adverse events occurred. CONCLUSIONS; In patients with severe PAH deteriorating despite ongoing prostanoid treatment, long-term adjunct oral sildenafil improves exercise capacity and pulmonary hemodynamics. A combination of prostanoids and sildenafil is an appealing concept for future treatment of pulmonary hypertension. PMID- 12849678 TI - Streptozotocin-induced hyperglycemia exacerbates left ventricular remodeling and failure after experimental myocardial infarction. AB - OBJECTIVES: The aim of the present study was to determine whether streptozotocin (STZ)-induced hyperglycemia exacerbates progressive left ventricular (LV) dilation and dysfunction after myocardial infarction (MI). BACKGROUND: Diabetes mellitus (DM) adversely affects the outcomes in patients with MI. However, it is unknown whether DM can directly affect the development of post-MI LV remodeling and failure. METHODS: Male mice were injected intraperitoneally with STZ (200 mg/kg; DM group) or vehicle only. At two weeks, MI was created in the STZ injected (DM+MI group) or vehicle-injected mice (MI group) by left coronary artery ligation, and they were followed up for another four weeks. RESULTS: Survival during six weeks was significantly lower in the DM+MI versus MI group (25% vs. 71%; p < 0.01), despite a similar infarct size (60 +/- 2% vs. 61 +/- 2%; p = NS). Echocardiography after two weeks of ligation showed LV dilation and dysfunction with MI, both of which were exaggerated in the DM+MI group. Likewise, LV end-diastolic pressure and lung weight were increased in mice with MI, and this increase was enhanced in the DM+MI group. The myocyte cross-sectional area in the non-infarcted LV increased to a similar degree in the DM+MI and MI groups, whereas the collagen volume fraction was greater in the DM+MI group. Deoxyribonucleic acid laddering was greater in the DM+MI group. CONCLUSIONS: Hyperglycemia decreased survival and exaggerated LV remodeling and failure after MI by increasing interstitial fibrosis and myocyte apoptosis. Diabetes mellitus could be a risk factor for heart failure, independent of coronary artery lesions. PMID- 12849679 TI - Induction of left ventricular remodeling and dysfunction in the recipient heart after donor heart myocardial infarction: new insights into the pathologic role of tumor necrosis factor-alpha from a novel heterotopic transplant-coronary ligation rat model. AB - OBJECTIVES: The present study investigated the effects of tumor necrosis factor (TNF)-alpha and angiotensin II (ANG II) on cardiac remodeling and dysfunction at the early stage of acute myocardial infarction (MI) by using a novel heterotopic cardiac transplantation-coronary ligation model. BACKGROUND: A recent clinical study has demonstrated a possible role of monocytosis in the development of left ventricular (LV) remodeling in patients with acute MI reperfusion. METHODS: We performed isogenic heterotopic cardiac transplantation and simultaneous coronary ligation to produce MI in the donor heart and to evaluate the hearts of both donors and recipients in Lewis rats. RESULTS: A significant decrease in LV fractional shortening and positive rate of rise in LV pressure and a significant increase in LV end-diastolic dimension/body weight and LV end-diastolic pressure were observed in the recipient hearts in the ligation group on day 7. TNF-alpha was significantly elevated not only in the plasma but also in the recipient hearts in the ligation group. In contrast, ANG II was significantly increased only in the infarct region of the donor hearts, but not in the plasma. Furthermore, the recipients' transient LV remodeling and dysfunction were completely abolished by the intravenous administration of a TNF-alpha antagonist. CONCLUSIONS; We developed a novel cardiac transplantation-coronary ligation model capable of inducing MI in the absence of downstream hemodynamic effects and allowing differential quantification of indexes of cardiac remodeling in vivo, including the local and remote effects of ANG II and TNF-alpha on cardiac remodeling. PMID- 12849680 TI - Prognostic significance of coronary blood flow velocity patterns in patients with reperfused acute myocardial infarction and TIMI-2 flow. PMID- 12849684 TI - Report by the editors. PMID- 12849685 TI - Recovery from manual metal arc-stainless steel welding-fume exposure induced lung fibrosis in Sprague-Dawley rats. AB - Welders with radiographic pneumoconiosis abnormalities have exhibited a gradual clearing of the X-ray identified effects following removal from exposure. In some cases, the pulmonary fibrosis associated with welding fumes appears in a more severe form in welders. Accordingly, to investigate the disease and recovery process of pneumoconiosis induced by welding-fume exposure, rats were exposed to welding fumes with concentrations of 63.6+/-4.1 mg/m(3) (low dose) and 107.1+/ 6.3 mg/m(3) (high dose) of total suspended particulate for 2 h per day in an inhalation chamber for a total of 2 h or 15, 30, 60 or 90 days. Thereafter, the rats were no longer exposed and allowed to recover from the welding fume-induced lung fibrosis for 90 days. When compared to the unexposed control group, the lung weights significantly increased in both the low- and high-dose rats from day 15 to 90. A histopathological examination combined with fibrosis-specific staining revealed that the lungs from the low-dose rats did not exhibit any significant progressive fibrotic changes. Whereas, the lungs from the high-dose rats exhibited early delicate fibrosis from day 15, which progressed into the perivascular and peribronchiolar regions by day 30. Interstitial fibrosis appeared at day 60 and became prominent by day 90, along with the additional appearance of pleural fibrosis. Recovery, evaluated based on the body and lung weights and a histopathological examination, was observed in both the high and low-dose rats that were exposed up to 30 days. The rats exposed for 60-90 days at the low dose also recovered from the fibrosis, yet the rats exposed for 60-90 days at the high dose did not fully recover. Consequently, recovery from pneumoconiosis induced by welding-fume exposure was observed when the degree of exposure was short-term and moderate. PMID- 12849686 TI - Assessment of pubertal development in juvenile male rats after sub-acute exposure to bisphenol A and nonylphenol. AB - The effects of bisphenol A and nonylphenol on pubertal development in the intact juvenile/peripubertal male Sprague-Dawley rats was observed in this study from PND23-52/53. Two groups of rats were administered orally with either 100 mg/kg body weight of nonylphenol or bisphenol A. Another group of rats were administered orally with a mixture of 100 mg/kg body weight of nonylphenol and bisphenol A. Control group was administered with the vehicle of Tween-80 with corn oil (1:9 v/v). Observations made in this study included growth, age at preputial separation, thyroid, liver, testis and kidney weight and histology, epididymal and seminal vesicle plus coagulation gland weight. Nonylphenol and bisphenol A have been observed to cause delay in puberty onset as well as testicular damage in the treatment groups when compared to the control; spermatogenesis was affected in most treated rats. Bisphenol A also caused the enlargement of the kidney and hydronephrosis. Administration of nonylphenol and bisphenol A as a mixture has caused less than additive effects. PMID- 12849687 TI - Estrous cyclicity and ovarian follicles in female rats after prenatal exposure to 3,3',4,4',5-pentachlorobiphenyl. AB - Alterations of estrous cyclicity and ovarian follicles following prenatal exposure to PCB126 were examined. Female SD rats were given (i.g.) 25 pg, 2.5, 250 ng and 7.5 microg of PCB126/kg or the vehicle on days 13-19 postconception. Vaginal opening (VO) in the 250 ng and 7.5 microg offspring was significantly delayed. All groups showed irregular estrous cyclicity following VO, but it became normal after a few days. However, the start of normal estrous cyclicity following VO in the 2.5, 250 ng and 7.5 microg groups was significantly delayed. At 30 and 50 days old, the 2.5, 250 ng and 7.5 microg groups showed significantly fewer antral follicles and a higher number of atretic follicles. The 7.5 microg group at 50 days old revealed significantly fewer corpus luteums. In 50-day-old offspring, the 2.5, 250 ng and 7.5 microg groups showed a significant reduction in serum 17beta-estradiol and progesterone levels and significantly higher levels of PCB126 in the fatty tissue compared with the vehicle group. Thus, while the prenatal dose of PCB126 used in this study did not induce malformation of the external genitalia or persistent ovarian disruption, disruption of ovarian function at puberty was found in the 2.5 ng group of pups born to dams exposed to 17.5 ng/kg PCB126. The present study suggests that PCB126, at least in part, exerted direct effects on the ovary as shown by the disruption of estrous cyclicity. PMID- 12849688 TI - Comparative evaluation of four urinary tubular dysfunction markers, with special references to the effects of aging and correction for creatinine concentration. AB - Comparative evaluation was made on alpha(1)-microglobulin (alpha(1)-MG), beta(2) microglobulin (beta(2)-MG), retinol binding protein (RBP) and N-acetyl-beta-D glucosaminidase (NAG), as a marker of renal tubular dysfunction after environmental exposure to cadmium (Cd), with special references to the effects of aging and correction for creatinine concentration. For this purpose, a previously established database of 817 never-smoking Japanese women (at the ages of 20 to 74 years) on hematological [hemoglobin, serum ferritin (FE), etc.] and urinary parameters [alpha(1)-MG, beta(2)-MG, creatinine (cr), and a specific gravity] was revisited. For the present analysis, the database was supplemented by the data on RBP and NAG in urine. The exposure of the women to Cd was such that the geometric mean Cd in urine was 1.3 microg/g cr. Among the four tubular dysfunction markers, NAG showed the closest correlation with Cd, followed by alpha(1)-MG and then beta(2)-MG, and RBP was least so although the correlations were all statistically significant. The observed values of the markers gave the best results, whereas correction for a urine specific gravity gave poorer correlation, and it was the worst when correction for creatinine concentration was applied. Age was the most influential confounding factor. The effect of age appeared to be attributable at least in part to the fact that both creatinine and, to a lesser extent, the specific gravity decreased as a function of age. Iron deficiency anemia of sub clinical degree as observed among the women did not affect any of the four tubular dysfunction markers. In conclusion, NAG and alpha(1)-MG, rather beta(2) MG or RBP, are more sensitive to detect Cd-induced tubular dysfunction in mass screening. The use of uncorrected observed values of the markers rather than traditional creatinine-corrected values is recommended when comparison covers people of a wide range of ages. PMID- 12849689 TI - Inhibition of aflatoxin B1 mutagenicity by cyclopiazonic acid in the presence of human liver preparations. AB - Co-occurrence of cyclopiazonic acid (CPA) and aflatoxin B(1) (AFB(1)) has been reported in different food commodities. Recently, we have shown that CPA reduces AFB(1) mutagenicity in the standard Salmonella-Microsome-Assay using rat S9-mix for metabolic activation (Environ. Toxicol. Pharmacol. 11 (2002) 207). When using S9-mix prepared from individual liver fractions of human patients, CPA was found to be non-mutagenic, but exerted a significant reduction of the mutagenicity of AFB(1). Moreover, CPA was shown to inhibit testosterone hydroxylation, but not methoxyresorufin dealkylation (MROD), in human S9. Thus, the reduction of the AFB(1) mutagenicity by CPA may be attributed to the inhibitory effect of CPA on cytochrome P450 (CYP450) 3A4 activity. These findings might be of relevance to the epidemiology of food-borne mycotoxicosis as similar molar ratios to those investigated here have been reported in food commodities. PMID- 12849690 TI - Antioxidant property of metallothionein in fasted mice. AB - Metallothionein (MT) is a low-molecular-weight and sulfur-rich protein that is induced by not only heavy metals but also physiological stresses such as fasting and restraint stresses. Although MT plays a role as a radical scavenger and a regulator of metabolism of metals, the biological function of MT induced by fasting stress has not been elucidated. In this study, we investigated the antioxidative role of MT in fasted mice. In fasted mice, the lipid peroxidation level of the liver was elevated by 24-h fasting stress, and pre-induction of hepatic MT by Zn diminished hepatic lipid peroxidation. Although 24-h fasting stress induced MT synthesis in the liver, other antioxidants such as catalase, manganese-superoxide dismutase (Mn-SOD), and glutathione peroxidase (GSHPx) were not activated in the liver. Moreover, the hepatic MT level was still elevated by fasting stress after seven cycles of repetition of alternate fasting and feeding every 24 h, but the activities of catalase, Mn-SOD and GSHPx were unchanged. These results indicate that MT induced by fasting stress plays partly as an antioxidant. PMID- 12849691 TI - Investigation of testicular toxicity of nefiracetam, a neurotransmission enhancer, in rats. AB - Testicular toxicity of nefiracetam (N-(2,6-dimethylphenyl)-2-(2-oxo-1 pyrrolidinyl) acetamide), a neurotransmission enhancer, was investigated in male Slc:SD rats. Nefiracetam was orally administered daily at 1500 mg/kg for 4 weeks, and the animals were killed sequentially during the course of administration to determine testicular histopathological changes and sperm head counts (SHC), and hormonal changes. Retention of step 19 spermatids, sporadic degeneration of pachytene spermatocytes and step 7 spermatids in the stage VII seminiferous tubules, and a decrease in SHC were seen as earliest changes after 1 week of administration. These changes gradually advanced up to atrophy of seminiferous tubules with multinucleated-giant-cell formation after 4-week administration. Serum and testicular testosterone levels were decreased, but recovered to the control levels within a day following a single administration, and the decreases were repeated after 1-week administration. These results suggest that nefiracetam induced earliest changes could be caused by the decreased level of testicular testosterone. PMID- 12849692 TI - Identification of organic fractions of diesel exhaust particulate (DEP) which inhibit nitric oxide (NO) production from a murine macrophage cell line. AB - Diesel exhaust particulates (DEPs) can constitute a large component of the particulate air pollution in urban areas and is a health concern. The effects of DEP on nitric oxide (NO) production by a murine macrophage cell line (RAW264.7) in response to interferon-gamma (INFgamma), lipopolysaccharide, (LPS) and Bacillus Calmette-Guerin (BCG) were studied. The DEP was fractionated into organic and inorganic fractions (carbonaceous core). The organic portion was further divided into asphaltene, saturates, less polar aromatics, more polar aromatics and resins-containing fractions. Each fraction was tested for the ability to suppress NO production from BCG-stimulated macrophages. DEP crude organic extract, more polar aromatic hydrocarbon, and resin fractions dose dependently inhibited BCG-stimulated NO production. It is concluded that the responsiveness of the macrophages to stimuli, such as BCG, is suppressed by DEP and that this activity is most predominant in the polar aromatic hydrocarbons and resins-containing fractions. PMID- 12849693 TI - Silica induces nuclear factor-kappaB activation through TAK1 and NIK in Rat2 cell line. AB - Silica has been known to be a factor in acute cell injury and chronic pulmonary fibrosis. In Rat2 fibroblasts, silica induced the activation of nuclear factor kappa B (NF-kappaB), which plays a crucial role in regulating the expression of many genes involved in the subsequent inflammatory response. In addition, we observed that transforming growth factor-beta activated kinase 1 (TAK1) and NF kappaB-inducing kinase (NIK) were involved in silica-mediated NF-kappaB activation in Rat2 cells. The dominant negative mutant forms of TAK1 and NIK inhibited the silica-induced NF-kappaB activation in Rat2 cells. Furthermore, we demonstrated that endogenous TAK1 is phosphorylated in silica-stimulated Rat2 cells. These results indicate that TAK1 functions as a critical mediator in the silica-induced signaling pathway. PMID- 12849694 TI - Study of the activity of several brain enzymes like markers of the neurotoxicity induced by perinatal exposure to lead and/or cadmium. AB - Five enzymes, considered keys to the correct function of the central nervous system (CNS) were chosen to study their utility as markers of the possible neurotoxic effects produced by the perinatal exposure to lead and/or cadmium. With this aim, lead acetate (300 mg/l) and/or cadmium acetate (10 mg/l) were administered to pregnant Wistar rats from day 1 of pregnancy to parturition (day 0) or until weaning (day 21). Brains were used to determine the activity of acid phosphatase (ACP), alkaline phosphatase (ALP), catalase (CAT), acetylcholinesterase (AchE) and ATP-ases (Na(+)/K(+) and Mg(2+)/Ca(2+) dependent). The general decrease of the activity of these enzymes observed after the treatment with heavy metals suggests that lead and cadmium are able to reach the CNS and impair its function by neurochemical changes. On the other hand, the results showed an increase of the activity of CAT; which possibly indicates that lead and cadmium might have other mechanisms of action, such as increasing oxidative damage in the CNS. Finally, the study of the effects produced by the simultaneous administration of lead and cadmium suggests that, in general, both metals exert an additive effect, either competing for the same inhibitory binding sites or increasing cellular damage. In conclusion, the study of the activity of enzymes is revealed as an effective method to measure the neurotoxicity induced by lead and cadmium. PMID- 12849695 TI - Ultraviolet-A induced changes in photosystem II of thylakoids: effects of senescence and high growth temperature. AB - Ultraviolet-A (UV-A) radiation induced changes in photosystem II (PS II) of senescing leaves of wheat seedlings were investigated. UV-A radiation did not show any significant effect on the level of photosynthetic pigments. However, the decline in F(v)/F(m) and oxygen evolution rate indicated the damaging effect of the radiation on primary photochemistry of PS II. Modification at the Q(B) binding site was inferred from the observed downshift of peak temperature of thermoluminescence (TL) B-bands. The UV-A induced changes in PS II of chloroplasts from senescing leaves were found to be synergistically accelerated by high growth temperature. PMID- 12849696 TI - Hydrogen-bonding dynamics of free flavins in benzene and FAD in electron transferring flavoprotein upon excitation. AB - The dynamic natures of two hydrogen-bonding model systems, riboflavin tetrabutylate (RFTB)-trichloroacetic acid (TCA) and RFTB-phenol in benzene, and of electron-transferring flavoprotein (ETF) from pig kidney upon excitation of flavins was investigated by means of steady state and time-resolved fluorescence spectroscopy. In both model systems fluorescence intensities of RFTB decreased as TCA or phenol was added. The spectral characteristics of ETF under steady state excitation were quite similar to those of the RFTB-TCA system, but not to those of the RFTB-phenol system. The observed fluorescence decay curves of ETF fit well with the calculated decay curves with two lifetime components, as in the model systems. Averaged lifetime was 0.9 ns. The time-resolved fluorescence spectrum of ETF shifted toward longer wavelength with time after pulsed excitation, which was also observed in the RFTB-TCA system. In the RFTB-phenol system the emission spectrum did not shift at all with time. These results reveal that the dynamic nature of ETF can be ascribed to aliphatic hydrogen-bonding(s) of the isoalloxazine ring with surrounding amino acid(s). From the fluorescence characteristics of ETF in comparison with the model systems, human ETF and other flavoproteins, it was suggested that ETF from pig kidney does not contain Tyr-16 in the beta subunit, unlike human ETF. PMID- 12849697 TI - Influence of substituted 1,4-anthraquinones on the chlorophyll fluorescence and photochemical activity of pea thylakoid membranes. AB - The effect of substituted 1,4-anthraquinones on the photochemical activity and chlorophyll fluorescence of thylakoid membranes was examined. Both the fluorescence and the photochemical activity depend on the 1,4-anthraquinone substituent. Stronger quinone-induced quenching of the chlorophyll fluorescence than quinone-induced changes in the activity of photosystem II is observed. The type (Cl or Br) and the position (Cl) of the chalogen atom strongly influence the degree of inhibition of PSII electron transport and the quenching of chlorophyll fluorescence. The data suggest that the quenching of chlorophyll fluorescence is due rather to the interaction of the 1,4-anthraquinones and chlorophyll molecules than to an indirect effect caused by stimulation of the photochemistry. PMID- 12849698 TI - Low-power laser irradiation improves histomorphometrical parameters and bone matrix organization during tibia wound healing in rats. AB - The influence of daily energy doses of 0.03, 0.3 and 0.9 J of He-Ne laser irradiation on the repair of surgically produced tibia damage was investigated in Wistar rats. Laser treatment was initiated 24 h after the trauma and continued daily for 7 or 14 days in two groups of nine rats (n=3 per laser dose and period). Two control groups (n=9 each) with injured tibiae were used. The course of healing was monitored using morphometrical analysis of the trabecular area. The organization of collagen fibers in the bone matrix and the histology of the tissue were evaluated using Picrosirius-polarization method and Masson's trichrome. After 7 days, there was a significant increase in the area of neoformed trabeculae in tibiae irradiated with 0.3 and 0.9 J compared to the controls. At a daily dose of 0.9 J (15 min of irradiation per day) the 7-day group showed a significant increase in trabecular bone growth compared to the 14 day group. However, the laser irradiation at the daily dose of 0.3 J produced no significant decrease in the trabecular area of the 14-day group compared to the 7 day group, but there was significant increase in the trabecular area of the 15 day controls compared to the 8-day controls. Irradiation increased the number of hypertrophic osteoclasts compared to non-irradiated injured tibiae (controls) on days 8 and 15. The Picrosirius-polarization method revealed bands of parallel collagen fibers (parallel-fibered bone) at the repair site of 14-day-irradiated tibiae, regardless of the dose. This organization improved when compared to 7-day irradiated tibiae and control tibiae. These results show that low-level laser therapy stimulated the growth of the trabecular area and the concomitant invasion of osteoclasts during the first week, and hastened the organization of matrix collagen (parallel alignment of the fibers) in a second phase not seen in control, non-irradiated tibiae at the same period. The active osteoclasts that invaded the regenerating site were probably responsible for the decrease in trabecular area by the fourteenth day of irradiation. PMID- 12849699 TI - Chemical and photolytical transformation of biomedically significant compounds in the presence of deuterated solvents. AB - The effect of the nature of solvent on the properties of biomedically important compounds is of particular importance. The conversion of certain biomedical compounds with deuterated solvents is an area of research that has not been accorded adequate recognition in the literature. We explored this area in the interest of shedding some light on the possible effects of solvent on the nature of the solute. The transformation of specific medically important compounds such as bilirubin, thymine, uracil, dehydrocholesterol (7-DHC) and vitamin D(3) was observed in the presence of deuterated solvents such as heavy water and deuterated chloroform. The products of the relevant reactions were confirmed spectrophotometrically. An additional feature to our investigation involved the photolysis of the aforementioned compounds by solar irradiation. The pure samples were dissolved in solutions of the deuterated solvents, corresponding to concentrations of typically 10(-2) mM, and exposed to sunlight for about 15-30 min. The deuterated solvents caused chemical transformation in all chemical compounds tested, and produced intense characteristic absorbance maxima between 200 and 700 nm. Sunlight exposure was also effective in either augmenting the effects of deuterated solvent as in bilirubin and 7-DHC or reducing it as with thymine or having no effect as with uracil or completely changing it as in vitamin D(3). It has been shown that the use of deuterated solvents produces unique chemical and photochemical conversions of bilirubin, 7-DHC, thymine, uracil and vitamin D(3). This was attributed to the fact that deuterated compounds display a somewhat different chemistry to their ordinary counterparts and that possibly thermodynamic considerations could be responsible for the novel transformations. PMID- 12849700 TI - Photodynamic inactivation of isolated crayfish neuron requires protein kinase C, PI 3-kinase and Ca2+. AB - Involvement of some signalling pathways in response to photodynamic therapy (PDT) of sulfonated aluminium phthalocyanine Photosens has been studied in isolated nerve cell. Neurone photosensitisation with 10(-7) M Photosens gradually inhibited firing and irreversibly abolished neuronal activity. Activation of protein kinase C (PKC) by phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) precipitated PDT-induced abolition of neurone activity and caused nucleus swelling and impairment of the nucleus border. Elevation of cytosolic Ca(2+) concentration by ionomycin or thapsigargin also reduced neurone lifetime. In contrast, the PKC inhibitors staurosporine, hypericin or chelerythrine as well as the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitors wortmannin or LY294002 increased neurone lifetime. These results showed that PKC, PI 3-kinase and Ca(2+) are involved in PDT-induced neurone inactivation and following death. PMID- 12849701 TI - Microfluorometric study of oxygen dependence of (1"-pyrene butyl)-2-rhodamine ester probe in mitochondria of living cells. AB - The access to oxygen concentration is of importance in various organelles of living cells, especially in mitochondria. A new probe, (1"-pyrene butyl)-2 rhodamine ester, was designed to target this organelle. We present here the properties of the probe in such an environment. Microspectrofluorometry confirms the localization of the probe in the mitochondrial environment at low doses (1 microM). At these doses, the cell toxicity experiments show no effect on the cell growth. The emission spectrum measured at an excitation wavelength of 340 nm (pyrene centered) indicates energy transfer from the pyrene to the rhodamine chromophore, as also observed in an ethanol solution. With excitation at 337 nm, the excited state decays biexponentially with lifetime decays of 6-9 ns and 90 ns. The first corresponds to the intrinsic fluorescence of the cell and the latter corresponds to the pyrene chromophore. In degassed conditions the pyrene lifetime decay increases up to 130 ns. Under an oxygen atmosphere the lifetime decays decrease to 62 ns. The lifetime changes with the oxygen concentration were compared with the range obtained during our previous study in ethanol solution (5 220 ns). The observed differences were interpreted by studying the lifetime of the probe in simplified environments, liposome suspensions and protein solutions. In this paper we show that the new probe can be used to measure the fluctuation of oxygen concentration in the surroundings of mitochondria. PMID- 12849702 TI - Conformation of gramicidin-A in CTAB micellar media. AB - Gramicidin A (gA) is a linear pentadecapeptide, which exhibits various conformations depending on the environment. The conformational behavior of gA in spherical and rod-shaped cationic micelles formed by cetyltrimethylammonium bromide (CTAB) surfactant has been studied using circular dichroism (CD) and fluorescence spectroscopy, and a probable structure of gramicidin A in CTAB media has been proposed. A CD study shows that gramicidin A assumes beta(6.3) helical structure in cationic spherical as well as rod-shaped CTAB micellar media. Modeling studies show the flexibility of the side chain conformation particularly in tryptophan-9. Study of intrinsic fluorescence of tryptophans in gramicidin A indicates three distinct environments for the four-tryptophan residues in CTAB media. PMID- 12849703 TI - Pre-exposure effects of 1 and 3 MHz therapeutic ultrasound on ConA activated spleenocytes. AB - This study was performed to evaluate the pre-exposure effects of ultrasound (1 MHz or 3 MHz) on ConA activated spleenocyte proliferation and cytokine production. Cells were treated for 10 min at various intensities, rested for 1h and stimulated with the T cell activator ConA. The cells were then analyzed for the effects of non-thermal ultrasound on cell growth and the presence of IL-2, IL 4 and IFN-g. The data show that pre-exposure of spleenocytes had no significant effects on the proliferation of ConA activated spleenocytes at either 1 or 3 MHz (10 min at 0.1 or 0.5 W/cm(2)). Significant increases in IL-2 were observed in both 1 and 3 MHz pre-treated and ConA activated spleenocytes. Cells pre-treated with 1 MHz and stimulated with ConA showed a significant increase in IL-4 and IFN g. Conversely, cells pre-treated with 3 MHz and stimulated with ConA show a significant decrease in IL-4 and IFN-g. Interleuken-4 is known to increase the growth of mast cells, inhibit macrophage activation and increases the activity of the T cell subpopulation, T(H2). Interferon-gamma is known to stimulate production of collagen in fibroblasts, enhance debridement activity of macrophage and inhibit activity of the T cell subpopulation, T(H2). PMID- 12849704 TI - A Bcl-2 transgene expressed in hepatocytes does not protect mice from fulminant liver destruction induced by Fas ligand. AB - We compared the biological mechanism of cell death during hepatotoxicity induced by ligation of the Fas receptor in wild-type and liver-specific bcl-2 transgenic mice. Transgenic overexpression of Bcl-2 in mouse hepatocytes can prevent lethal hepatitis induced by agonistic anti-Fas antibodies. In contrast, Fas ligand (FasL)-induced death cannot be overcome in bcl-2 transgenic mice, indicating that anti-Fas antibodies do not reliably mimic the more physiological ligand. Different apoptotic parameters, viz. caspase activation, cytochrome c release and nuclear DNA degradation were analysed. No differences, however, could be observed between wild-type and bcl-2 transgenic mice after injection with a lethal dose of soluble FasL, indicating that apoptosis by FasL-dependent ligation is not modulated by Bcl-2 in vivo. These results demonstrate that the stimulus determines the outcome between type I mitochondria-independent apoptosis, in the case of FasL, or type II mitochondria-dependent and Bcl-2-inhibitable apoptosis, in the case of anti-Fas antibodies. PMID- 12849705 TI - Hypoglycaemia downregulates endotoxin-induced production of tumour necrosis factor-alpha, but does not affect IL-1beta, IL-6, or IL-10. AB - The aim of the present study was to investigate the effect of hypoglycaemia on the production capacity of the proinflammatory cytokines tumour necrosis factor alpha (TNFalpha) and interleukin-1beta (IL-1beta) in subjects with and without diabetes. Hyperinsulinaemic (360 pmolm(-2) x min(-1)) stepped hypoglycaemic (5.0 3.5-2.5 mmoll(-1)) glucose clamps were performed in eight diabetic patients and in six non-diabetic subjects, and hyperinsulinaemic normoglycaemia (5.0 mmoll( 1)) control experiments were performed in four non-diabetic subjects. Circulating levels of cytokines and endotoxin-induced production of TNFalpha, IL-1beta, IL-6, and IL-10 were assessed. The effects of insulin and adrenaline were measured in separate in vitro experiments. In non-diabetic subjects, hypoglycaemia downregulated the production capacity of TNFalpha in a concentration-dependent fashion (P=0.007), but not of IL-1beta, IL-6, or IL-10. Compared to controls, the production capacity of TNFalpha in diabetic patients was already suppressed at normoglycaemia (P=0.02) and only fell in response to hypoglycaemic nadir (P=0.04). The downregulation of TNFalpha could not be explained by increased insulin or adrenaline levels. We conclude that hypoglycaemia specifically downregulates TNFalpha production capacity. Diabetic patients already have a suppressed TNFalpha production capacity at non-hypoglycaemic levels. PMID- 12849706 TI - Glutamine decreases lipopolysaccharide-induced IL-8 production in Caco-2 cells through a non-NF-kappaB p50 mechanism. AB - Glutamine (Gln) supplementation has been shown to decrease production of pro inflammatory cytokines by the human intestinal mucosa. The mechanism of this is poorly understood. We hypothesize that Gln down-regulates lipopolysaccharide (LPS)-stimulated pro-inflammatory cytokine production in Caco-2 cells by nuclear factor-kappa B (NF-kappaB). Caco-2 cells were incubated with different concentrations of Gln with or without methionine sulfoximine (MS, an inhibitor of glutamine synthetase) before stimulation with LPS. IL-6, IL-8, IL-10 and TNF alpha protein and mRNA level were determined. NF-kappaB translocation was determined using an ELISA-based kit. IL-8 was the only detectable cytokine/chemokine. The largest amount of IL-8 was secreted by cells in the presence of MS with no Gln in the medium after exposure to LPS. LPS increased IL 8 production, peaking 10h after LPS administration. The addition of Gln (0.5 or 5.0mM) decreased IL-8 peptide and mRNA expression. LPS increased NF-kappaB nuclear translocation in the presence or absence of MS. Neither Gln nor MS altered NF-kappaB nuclear translocation. These results indicate that the lack of glutamine increases IL-8 production by Caco-2 cells after LPS stimulation. However, the glutamine-mediated decrease in LPS-stimulated IL-8 production is not associated with NF-kappaB p50 nuclear binding. PMID- 12849707 TI - Cytokine gene expression--part of host defence in pulpitis. AB - Analyses of cytokines mediating inflammatory reactions are key to understanding the etiopathology of various diseases. This study investigated differences in cytokine gene expression between pulps from healthy virgin teeth and from symptomatic vital teeth with severe caries lesions in a group of young, healthy individuals. The mRNA levels of IL-1alpha, IL-1beta, IL-6, IL-8, and IL-18 were measured concomitantly by quantitative real-time RT-PCR. IL-1alpha and IL-1beta were not expressed at significantly higher levels in symptomatic versus clinically healthy pulps, while the difference was significant for the other cytokines (log-rank test, P<0.05). A concordance test for independence revealed significant correlation between IL-1alpha and IL-1beta, and between IL-6, IL-8, and IL-18 mRNA levels (P<0.05). The cytokine-specific differences revealed a differential significance of gene expression in cytokine regulation. The hypothesis that increase of cytokine mRNA expression is part of host reaction in pulpitis was corroborated by our observation. PMID- 12849708 TI - In situ localization of TNFalpha/beta, TACE and TNF receptors TNF-R1 and TNF-R2 in control and LPS-treated lung tissue. AB - Tumor necrosis factor (TNF) has been implicated in several infectious and inflammatory lung diseases. Two closely related variants, TNFalpha and TNFbeta, elicit various cellular responses via two distinct TNF receptors, the 55-kDa TNF R1 and the 75-kDa TNF-R2. Recently, a TNFalpha-converting enzyme (TACE) was described, which cleaves and releases the membrane-bound TNFalpha. In the present study in normal rat and human lung tissue, the constitutive expression of TNFalpha/beta, TACE and TNF-R1/R2 was investigated by immunohistochemical techniques. In addition, TNFalpha and TNFbeta mRNA were localized by in situ hybridization. Both TNFalpha and TNFbeta were detected in various lung cell types. Expression of TNFalpha was particularly prominent in bronchial epithelial cells and vascular smooth muscle cells, next to alveolar macrophages. Both in situ hybridization for TNFalpha message and TACE immunostaining matched this expression profile. TNFbeta-so far only known to be produced by lymphocytes-was demonstrated in alveolar macrophages, bronchial epithelial cells, vascular smooth muscle cells and endothelial cells at the protein and the message level. Both TNF receptors were detected, with TNF-R1 being prominent on bronchial epithelial cells and endothelial cells, and TNF-R2 being expressed by nearly all cell types. Following LPS stimulation in isolated rat lungs TNFalpha/beta signal intensity was largely reduced due to liberation of stored TNFalpha/beta, while TACE immunoreactivity remained unchanged or was enhanced, demonstrating increased TNF generation. We conclude that both TNFalpha and TNFbeta are constitutively expressed by several non-leukocytic cell types in the human and rat lung. In concert with the expression of TACE and the TNF receptors R1 and R2, this finding suggests in addition to the known role of the TNF system in inflammation physiological functions of the TNF system in different compartments of the adult lung, with the vasculature and the bronchial tissue being of particular interest in addition to the leukocyte/macrophage populations. PMID- 12849709 TI - Expression of cytokines by multipotent neural progenitor cells. AB - Recent work with mammalian neural stem cells has highlighted the role of cytokine signaling in the proliferation and differentiation of these multipotent cells. While the responsiveness of neural progenitors to exogenously applied growth factors has been demonstrated in vivo as well as in vitro, little attention has been given to the production of cytokines by these cells. Here we use immunocytochemistry, RT-PCR, and ELISA to show that under standard growth conditions multipotent neural progenitor cells from humans express multiple cytokines including IL-1alpha, IL-1beta, IL-6, TGF-beta1, TGF-beta2, TNF-alpha, but not IL-2, IL-4, or IFN-gamma. Neural progenitor cells from rat and mouse express some, but not all, of these cytokines under similar conditions. While the function of cytokine expression by neural progenitor cells remains to be elucidated, these signaling molecules are known to be involved in neural development and may play a role in the activation of quiescent stem cells by a variety of pathological processes. PMID- 12849710 TI - Chronic pain and fibromyalgia: the regulatory perspective. AB - Chronic pain is an important clinical entity that represents a currently unmet medical need. Relief of pain is an important public health goal for patients of all ages, from perinatal to geriatric. This article will describe some of the current regulatory issues in developing and approving drugs to treat chronic pain. It will also begin to familiarize the reader with the importance of the so called 'label' and some of its roles to enable the best 'risk-benefit' decisions be made for, and by, patients with chronic pain. PMID- 12849711 TI - The epidemiology of chronic generalized musculoskeletal pain. AB - Chronic widespread musculoskeletal pain has been subjected to several epidemiological studies during the last decade. According to these, approximately 10% of the general population report such complaints, clearly indicating chronic widespread musculoskeletal pain as a major health problem in the Western world. Almost unanimously, all studies found higher rates of such complaints among women compared with men, but the mechanisms responsible for the skewed gender ratio remain unknown.Chronic widespread musculoskeletal pain is the clinical hallmark of fibromyalgia and has been the subject of numerous epidemiological studies. The prevalence of fibromyalgia is reportedly 3-5%, again with a significant female predominance. Although the aetiopathogenesis of both fibromyalgia and chronic widespread musculoskeletal pain without other features of fibromyalgia remains an enigma, there is a body of evidence suggesting psychological and sociocultural factors as important for contracting such pain syndromes. PMID- 12849712 TI - Chronic diffuse musculoskeletal pain, fibromyalgia and co-morbid unexplained clinical conditions. AB - This chapter reviews our current knowledge on the presence of overlapping syndromes in one form of chronic diffuse pain, fibromyalgia. Patients with fibromyalgia often present with signs and symptoms of other unexplained clinical conditions, including chronic fatigue syndrome, irritable bowel syndrome, temporomandibular disorders, and multiple chemical sensitivities. The high prevalence, impact on function and opportunities for treatment underscore the need for clinicians and researchers to screen routinely for co-morbid unexplained clinical conditions among persons with fibromyalgia. We, therefore, describe a simple approach to screening for such conditions in accordance with published criteria. Interventions should directly address both fibromyalgia symptoms and co morbid unexplained clinical conditions, as well as the multiple factors that propagate pain, fatigue and limitations in function. PMID- 12849713 TI - Measuring clinical pain in chronic widespread pain: selected methodological issues. AB - Assessing clinical pain is an important task in clinical practice and research. A large empirical literature has documented that patients' pain reports can be systematically biased by a number of methodological factors. This chapter reviews a selection of methodological issues that can affect pain ratings, including: the impact of recall bias, the use of paper and electronic diaries to assess pain experiences, ecological momentary assessment methods as a way of capturing real time pain data in the real world, and pain scaling and data analysis considerations. Data from a recent study that implemented an electronic diary for capturing real-time pain data are presented and reviewed in the context of the methodological factors reviewed above. It is concluded that methodological factors can greatly affect our understanding of patients' pain experiences. PMID- 12849714 TI - Evoked pain measures in fibromyalgia. AB - Fibromyalgia is defined by widespread pain and tenderness at a minimum of 11 of 18 defined tender points. Current evidence indicates that tender points are not unique to fibromyalgia and are simply regions in the body where all people are more tender. Tenderness (i.e. sensitivity to pressure) is widespread in fibromyalgia rather than being confined to tender points, and patients are also more sensitive to heat, cold and electrical stimulation. Using the number of painful tender points as a measure of tenderness is clinically expedient but is theoretically vulnerable to bias and is influenced by subjective distress. Other means of assessing tenderness (e.g. pressure dolorimeter devices, or more elaborate psychophysical methods) demonstrate the same increased pain sensitivity in fibromyalgia that is noted with tender point assessments, but these measures are relatively independent of biasing factors or distress. Fibromyalgia is one of only a few syndromes defined by the presence of both spontaneous (i.e. clinical) and evoked (i.e. experimental) pain. While the issues associated with the evaluation of spontaneous pain are shared with all chronic pain syndromes, the issues associated with the evaluation of evoked pain sensitivity are specific to fibromyalgia and related musculoskeletal disorders. This chapter focuses on the evaluation of altered pain sensitivity in fibromyalgia. It describes current measurement methodology, briefly reviews studies of sensitivity to experimentally evoked painful and non-painful sensations, analyses the factors assessed by different measurement methodologies, and concludes with recommendations for future diagnostic criteria and measurement methods. PMID- 12849715 TI - Pharmacological therapies in fibromyalgia. AB - The fibromyalgia syndrome (FMS) is a common, chronic, widespread pain disorder that mainly affects middle-aged women. In addition to pain complaints, fatigue and disturbed sleep are symptoms frequently reported by these patients. Many FMS patients also meet diagnostic criteria for mood disorders (e.g. depression) as well as other so-called 'functional somatic syndromes', including irritable bowel syndrome, temporomandibular joint disorder, and subsets of chronic low-back pain. A wide variety of medications are used to manage the eclectic symptomatology of FMS patients, although relatively few have been rigorously tested. This chapter provides a contemporary update of the state of FMS pharmacotherapy, with an emphasis on compounds that have been tested in double-blind, randomized, controlled trials. Particular attention is paid to the efficacy of these therapies on the associated symptoms and co-morbid syndromes commonly seen in FMS patients. PMID- 12849716 TI - Physical exercise in fibromyalgia and related syndromes. AB - Fibromyalgia and related syndromes are characterized by chronic pain and fatigue. This chapter identifies the types of exercise that are effective for these patients and provides recommendations for exercise prescriptions. Based on a systematic review of randomized controlled studies of exercise, we suggest that low-intensity aerobic exercise, such as walking, can improve function and symptoms. Aerobic exercise performed twice a week at moderate intensity can improve aerobic capacity and reduce tenderness. Pool exercise can improve function, distress and symptoms. Strength training at adequate load can improve strength without exacerbation of symptoms. Most patients tolerate low-intensity exercise. High-intensity exercise should be undertaken with caution. Due to the large variability of functioning and symptom severity in patient populations, exercise prescriptions should be individualized and should include a long-term plan to maximize functioning and wellbeing. Studies with larger populations, allowing subgroup analyses regarding benefits and adverse effects of programmes, are needed. PMID- 12849717 TI - Psychological and behavioural therapies in fibromyalgia and related syndromes. AB - Psychological and behavioural therapies are being applied to patients with fibromyalgia (FM) with increasing frequency. The rationale for including psychological therapies is not for the treatment of co-morbid mood disorders, but rather to manage the many non-psychiatric psychological and social factors that comprise pain perception and its maintenance. This chapter reviews the involvement of mental health professionals under both the biomedical and biopsychosocial models of illness and describes cognitive behavioural therapy (CBT), a commonly used form of psychological therapy in the management of chronic pain conditions. The empirical literature supports the use of CBT with FM in producing modest outcomes across multiple domains, including pain, fatigue, physical functioning and mood. Greatest benefits appear to occur when CBT is used adjunctively with exercise. While the benefits are not curative or universally obtained by all patients, the benefits are sufficiently large to encourage future refinement of CBT for this population of patients. PMID- 12849718 TI - Complementary and alternative medicine in fibromyalgia and related syndromes. AB - Complementary and alternative medicine (CAM) has gained increasing popularity, particularly among individuals with fibromyalgia syndrome (FMS) for which traditional medicine has generally been ineffective. A systematic review of randomized controlled trials (RCTs) and non-RCTs on CAM studies for FMS was conducted to evaluate the empirical evidence for their effectiveness. Few RCTs achieved high scores on the CONSORT, a standardized evaluation of the quality of methodology reporting. Acupuncture, some herbal and nutritional supplements (magnesium, SAMe) and massage therapy have the best evidence for effectiveness with FMS. Other CAM therapies have either been evaluated in only one RCT with positive results (Chlorella, biofeedback, relaxation), in multiple RCTs with mixed results (magnet therapies), or have positive results from studies with methodological flaws (homeopathy, botanical oils, balneotherapy, anthocyanidins, dietary modifications). Lastly, other CAM therapies have neither well-designed studies nor positive results and are not currently recommended for FMS treatment (chiropractic care). PMID- 12849719 TI - Chronic widespread pain and fibromyalgia: what we know, and what we need to know. AB - Fibromyalgia (FM) is currently defined as the presence of both chronic widespread pain (CWP) and the finding of 11/18 tender points on examination. Only about 20% of individuals in the population with CWP also have 11/18 tender points; these individuals are considerably more likely to be female, and have higher levels of psychological distress. There is no clear clinical diagnosis for the other 80% of individuals with less than 11/18 tender points, but it is likely that these persons, like FM patients, also have pain that is 'central' (i.e. not due to inflammation or damage of structures) rather than peripheral in nature. Research into FM has taught us a great deal about the confluence of neurobiological, psychological and behavioural factors that can cause chronic central pain. These conditions respond best to a combination of symptom-based pharmacological therapies, and non-pharmacological therapies such as exercise and cognitive behavioural therapy. In contrast to drugs that work for peripheral pain due to damage or inflammation (e.g. NSAIDs, corticosteroids), neuroactive compounds [especially those that raise central levels of noradrenaline (norepinephrine) or serotonin] are most effective for treating central pain. PMID- 12849721 TI - Cytotoxicity of inorganic mercury in murine T and B lymphoma cell lines: involvement of reactive oxygen species, Ca(2+) homeostasis, and cytokine gene expression. AB - Mercury is a highly toxic heavy metal; exposure to mercury in humans and animals causes damage in several organs or systems including the immune system. To characterize the toxicity of mercury in the immune cells, the cytotoxic effects of inorganic mercury were studied in two distinct lymphoma lines, the murine T lymphoma (EL4) and B lymphoma (A20) cells. Mercury concentration-dependently decreased cell viability, membrane integrity, and proliferation in both EL4 and A20 cells. Mercury increased the reactive oxygen species (ROS) production in both EL4 and A20 cells, and pretreatment with antioxidants reversed mercury-induced ROS generation. Pretreatment of cells with antioxidants N-acetylcysteine (NAC) and silymarin decreased mercury-induced lactate dehydrogenase (LDH) release in both types of cells; however, Ca(2+) channel blocker lanthanum (La(2+)) decreased it only in A20 cells. The mode of cytotoxicity was a mixture of both apoptosis and necrosis. Mercury-induced apoptosis and necrosis in the two cell lines were indicated by staining with Hoechst 33258, propidium iodide, and co-staining with annexin V and propidium iodide. Both mercury-induced apoptosis and necrosis were attenuated by antioxidants. Mercury increased gene expression of IL-4 and TNFalpha in EL4 cells; these cytokines were not expressed in A20 cells. Data suggested different pathways of mercury-induced cytotoxicity in T and B lymphoma cells and involvement of ROS, Ca(2+) homeostasis, and inflammatory cytokine gene expression. PMID- 12849722 TI - Comparative toxicity of ifosfamide metabolites and protective effect of mesna and amifostine in cultured renal tubule cells. AB - Renal injury is a common side effect of the chemotherapeutic agent ifosfamide. Current evidence suggests that the ifosfamide metabolite chloroacetaldehyde contributes to this nephrotoxicity. The present study examined the effects of chloroacetaldehyde and acrolein, another ifosfamide metabolite, on rabbit proximal renal tubule cells in primary culture. The ability of the uroprotectant medications sodium 2-mercaptoethanesulfonate (mesna) and amifostine to prevent chloroacetaldehyde- and acrolein-induced renal cell injury was also assessed. Chloroacetaldehyde and acrolein (25-200 M) produced dose-dependent declines in neutral red dye uptake, glucose transport and glutathione content. Chloroacetaldehyde was a more potent toxin than acrolein. Pretreatment of cells with the glutathione-depleting agent buthionine sulfoximine enhanced the toxicity of both chloroacetaldehyde and acrolein while co-administration of mesna or amifostine prevented metabolite toxicity. These results support the hypothesis that chloroacetaldehyde is responsible for ifosfamide-induced nephrotoxicity. The protective effect of mesna and amifostine in vitro contrasts with clinical experience showing that these medications do not eliminate ifosfamide nephrotoxicity. PMID- 12849723 TI - Mutagenicity of bitumen and asphalt fumes. AB - The mutagenicity of asphalt fumes was tested with the Salmonella bioassays. The aim was to investigate if recycled additives modify the genotoxicity of emissions. Recycling of old asphalt is increasing, and we studied also the mutagenicity of emissions sampled during the re-use of asphalt. The composition of vapours and fumes were analysed by gas chromatography and by liquid chromatography. Bitumens containing coal fly ash (CFA) or waste plastics were heated to the paving temperatures in the laboratory. In the field, bitumen fumes were collected during paving of stone mastic asphalts (lime or CFA as a filler), remixing of stone mastic asphalt (lime or CFA as a filler), and of asphalt concrete. All the lab-generated vapour fractions were non-mutagenic. The particulate fractions were mutagenic with TA98 in the presence of the S9 activation. In addition, the lab-fumes from bitumen containing waste plastics were positive with both strains without S9. Only particulate fractions sampled in the field were tested. They were mutagenic with and without metabolic activation with both strains. The mutagenic potency of the field samples was higher than that of the lab-generated fumes without S9, and the remixing fumes were more mutagenic than the normal paving and lab-generated fumes with S9. The use of inorganic additive, CFA, did not change the mutagenicity of the fumes, whereas the organic additive, waste plastics, increased the mutagenicity of the laboratory emissions significantly. PMID- 12849724 TI - Aromatase activity modulation by lindane and bisphenol-A in human placental JEG-3 and transfected kidney E293 cells. AB - Aromatase is the cytochrome P-450 involved in converting androgens to estrogens. The cytochrome P-450 family plays a central role in the oxidative metabolism of compounds including environmental pollutants. Since lindane and bisphenol-A (BPA) are two well-characterized endocrine disruptors that have been detected in animals and humans, it was important to learn whether they could affect aromatase activity and consequently estrogen biosynthesis. The present study investigates the effects of BPA and lindane on cytotoxicity, aromatase activity and mRNA levels in human placental JEG-3 cells and transfected human embryonal kidney 293 cells. Both cell lines were exposed to increasing concentrations of lindane (25, 50 and 75 microM) and bisphenol-A (25, 50 and 100 microM) over different time periods (10 min-18 h). As a result, none of these concentrations showed cytotoxicity. After short pre-incubation times (10 min-6 h), aromatase activity was enhanced by both compounds. Longer time incubation (18 h), however, produced dose-related inhibition. Lindane and BPA had no significant effects on CYP19 mRNA levels. Therefore, lindane and BPA modulate aromatase activity suggesting an interaction with the cytochrome P-450 aromatase. This study highlights the endocrine-modulating properties of lindane and bisphenol-A. PMID- 12849725 TI - The role of quinone reductase (NQO1) and quinone chemistry in quercetin cytotoxicity. AB - The effects of quercetin on viability and proliferation of Chinese Hamster Ovary (CHO) cells and CHO cells overexpressing human quinone reductase (CHO+NQO1) were studied to investigate the involvement of the pro-oxidant quinone chemistry of quercetin. The toxicity of menadione was significantly reduced in CHO+NQO1 cells compared to wild-type CHO cells, validating the NQO1-overexpression in the CHO+NQO1 transfectant. Quercetin inhibited the proliferation of wild-type CHO and CHO+NQO1 cells to a similar extent without affecting cell viability, indicating that NQO1 enrichment of CHO cells did not provide increased protection. On the other hand, inhibition of NQO1 in both types of cells by dicoumarol significantly potentiated the inhibitory effect of quercetin on cell proliferation, revealing the role of NQO1 in cellular protection against quercetin. Altogether, these results can be explained by the hypothesis that both wild-type CHO and CHO+NQO1 cells contain sufficient NQO1 activity for optimal protection against the pro oxidant effect of quercetin on cell proliferation. The results also point at a cellular NQO1 threshold for optimal protection against quercetin. This NQO1 threshold seems to be in the range of NQO1 activities already present in various tissues. PMID- 12849726 TI - Loss of mitochondrial transmembrane potential and caspase-9 activation during apoptosis induced by the novel styryl-lactone goniothalamin in HL-60 leukemia cells. AB - Styryl-lactones such as goniothalamin represent a new class of compounds with potential anti-cancer properties. In this study, we investigated the mechanisms of goniothalamin (GTN), a plant styryl-lactone induced apoptosis in human promyelocytic leukemia HL-60 cells. This plant extract resulted in apoptosis in HL-60 cells as assessed by the externalisation of phosphatidylserine. Using the mitochondrial membrane dye (DIOC(6)) in conjunction with flow cytometry, we found that GTN treated HL-60 cells demonstrated a loss of mitochondrial transmembrane potential (Deltapsi(m)). Further immunoblotting on these cells showed activation of initiator caspase-9 and the executioner caspases-3 and -7. Pretreatment with the pharmacological caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone (Z-VAD.FMK) abrogated apoptosis as assessed by all of the apoptotic features in this study. In summary, our results demonstrate that goniothalamin induced apoptosis occurs via the mitochondrial pathway in a caspase dependent manner. PMID- 12849727 TI - Effects of the mycotoxin ochratoxin A and some of its metabolites on human kidney cell lines. AB - The modulations of complement-regulating surface proteins on a human embryonic and a renal carcinoma cell line are described regarding the effects of ochratoxin A and some of its metabolites on the surface markers CD46, CD55 and CD59. Membrane integrity, cell proliferation and metabolic activity were reduced to different extents, depending on the kind of mycotoxin and the dosage, which was ranging from 10 to 1000 ng/ml. The number of cells carrying surface markers was suppressed significantly at 1000 ng/ml, in some cases even at 100 ng/ml, whereas the intensity of receptor expression on the positive cells was found to be stimulated. The fraction RE2 (OTC) isolated from an OTA-containing crude toxin surpassed the effects of all other ochratoxin metabolites. Apart from well-known cytotoxic and genotoxic effects modulation of cell surface marker expression by low concentrations of OTA and OTC deserves more attention with regard to its immuno-pathogenic importance. Furthermore, occurrence and impact of the mycotoxin OTC should be studied more into detail. PMID- 12849728 TI - Predictive value of comparative molecular field analysis modelling of naphthalene inhibition of human CYP2A6 and mouse CYP2A5 enzymes. AB - The objects of this study were first to compare how well the recently constructed structure-inhibition activity relationship models of mouse CYP2A5 and human CYP2A6 predict the interaction of naphthalene in liver microsomes and secondly to study if these CYP enzymes actually oxidize naphthalene. The CoMFA model of CYP2A5 predicted the IC(50) value of naphthalene to be 42 microM (18-115 microM 95% CL) whereas in the in vitro experiment the result was 74 microM (65-83 microM) with the corresponding values for CYP2A6 being 41 microM (18-112 microM) and 25 microM (21-30 microM), respectively. Naphthalene appeared to be a competitive inhibitor both for mouse and human liver microsomal coumarin 7 hydroxylase, which is the specific probe activity for CYP2A5 and CYP2A6. The K(i) value for the mouse enzyme was between 12-26 microM and for the human enzyme 1.2 5.6 microM. A 1-h in vitro incubation of naphthalene with human and pyrazole treated mouse liver microsomes produced more 1-naphthol than 2-naphthol. Antibody against the purified CYP2A5 inhibited 50-60% of the formation of 1-naphthol and 30-40% of the formation of 2-naphthol. These results indicate that in silico CoMFA models predict relatively well the interaction of naphthalene with CYP2A5 and CYP2A6 and that these CYPs actually oxidize naphthalene in vitro. CoMFA CYP2A5 and CYP2A6 models are thus useful as a technique for elucidating the interaction and potency of untested chemicals with these CYPs. PMID- 12849729 TI - Induction of cytotoxicity and ssDNA breaks by 9-bromo-5-morpholino-tetrazolo[1,5 c]quinazoline in tumor cells cultured in vitro. AB - 9-Bromo-5-morpholino-tetrazolo[1,5-c]quinazoline (BMTQ) acted cytotoxically on murine leukemia cell line L1210 and human colon carcinoma cells Caco-2. We found the two highest concentrations of BMTQ (149.2 and 74.6 microM) induced an acute cytotoxic effect, however other tested concentrations (<74.6 microM) manifested a concentration/dependent and time/dependent cytotoxic effect. The sensitivity of murine leukemia cells L1210 and human colon carcinoma cells Caco-2 was expressed in the same order. The cytotoxicity of BMTQ was not accompanied by changes of the cell cycle profile. Following the cytotoxicity-related effects of BMTQ we observed the induction of ssDNA breaks after BMTQ treatment. All the concentrations of BMTQ increased the level of ssDNA breaks 1.3-2.9 times (after 2 h of treatment) and 1.6-2.8 times (after 4 h of treatment) in Caco-2 cells compared to the control. No apoptotic DNA fragmentation induced by BMTQ in Caco-2 cells was recorded. PMID- 12849730 TI - Anticlastogenic effect of aqueous extracts of Agaricus blazei on CHO-k1 cells, studying different developmental phases of the mushroom. AB - The Agaricus blazei Murill (ABM) mushroom, known as the sun mushroom, is native to Brazil and has become known for its medicinal properties. This study examined the anticlastogenic effect of Agaricus blazei in Chinese hamster ovary cells, CHO k1, by means of a chromosome aberration test using methyl methanesulphonate (MMS, 10(-4)M) as the DNA damage inducing agent. Two mushroom lines were used, ABM 99/26 and ABM 97/11, and the latter was used in the young (Y) and sporulating (S) developmental phases. The cells were treated for 12 h with MMS alone or combined with aqueous extracts of A. blazei at a final concentration of 0.15%, which were prepared at three different temperatures: (a) hot (60 degrees C), (b) room temperature (25 degrees C) and (c) chilled (4 degrees C). Mushroom extracts showed a marked anticlastogenic effect against DNA damage, as evidenced by a decrease in the number of cells with breaks, regardless of the line used, or the developmental stage or the temperature at which the extract was prepared. Generally, the extracts were more effective in reducing the isochromatid type breaks. The data obtained suggest that extracts of A. blazei mushroom are anticlastogenic under the conditions tested, mainly during the G1 and S stages of the cell cycle, where chromosome breaks of the isochromatid type are produced by the MMS agent. PMID- 12849731 TI - An assessment of the phototoxic hazard of a personal product ingredient using in vitro assays. AB - Where substances are intended for use in personal products applied to the skin an assessment of potential phototoxic hazard is required. This report describes a tiered testing strategy involving in vitro assays used for the phototoxic hazard assessment of a personal product ingredient (Ingredient X). The initial assay was measurement of a UV/visible absorption spectrum to identify absorption at relevant wavelengths. This was followed by in vitro assays for phototoxicity (3T3 cell neutral red uptake phototoxicity test) and photoallergy (photobinding to human serum albumin). These in vitro screens gave equivocal results for Ingredient X which appeared to suggest a weak phototoxic reaction. To further evaluate the phototoxic hazard of Ingredient X to human skin, a phototoxicity assay using a 3-D human skin model was conducted. Ingredient X did not cause phototoxicity in this assay. Overall conclusions from these studies were that although Ingredient X showed slight intrinsic potential for photoactivation, it was unlikely to present a hazard to human skin. This report illustrates the value in a step-wise strategy of the use of human skin models to help interpret the results of other in vitro phototoxicity assays. PMID- 12849732 TI - Rates of metabolism of chlorzoxazone, dextromethorphan, 7-ethoxycoumarin, imipramine, quinidine, testosterone and verapamil by fresh and cryopreserved rat liver slices, and some comparisons with microsomes. AB - In the present study we have investigated the disappearance of chlorzoxazone, dextromethorphan, 7-ethoxycoumarin, imipramine, quinidine, testosterone and verapamil from the medium in which fresh and cryopreserved rat liver slices were incubated. These compounds are all substrates of major isoforms of cytochrome P450 expressed in the liver. The metabolism of five of these compounds in microsomes from rat liver was also examined. Determinations of the concentrations of the compounds were performed employing LC/MS. Intrinsic clearance values (CL(ints)) were calculated on the basis of the concentration-vs.-time curves. No significant differences in the CL(int) values obtained with fresh and cryopreserved rat liver slices were observed for any of the compounds. The highest CL(int) value estimated with liver slices was observed for testosterone and the lowest values were with chlorzoxazone and 7-ethoxycoumarin. The total CL(int) values for 7-ethoxycoumarin and imipramine, calculated using scaling factors, were similar for liver slices and microsomes. In the case of testosterone, this total CL(int) was approximately 3.7-fold lower, whereas for dextromethorphan and quinidine it was 2.5- and 8.5-fold higher, respectively, with liver slices than with microromes. In conclusion, the rate of metabolism of the seven compounds tested with rat liver slices was not affected by cryopreservation. This finding adds further support to the general conclusion that the major activities involved in drug metabolism are not affected by cryopreservation of rat liver slices. PMID- 12849733 TI - Verification of the BALB/c 3T3 cell transformation assay after improvement by using an ITES-medium. AB - We carried out the first-step verification study on our ITES-medium-improved BALB/c 3T3 cell transformation assay. In order to estimate its potential use as a short-term screening method for putative carcinogens, 31 chemicals were tested in the improved transformation assay. The test chemicals consisted of 18 carcinogens and 13 noncarcinogens. The present improved transformation assay did not use an exogenous metabolizing system. Data analysis was carried out on 34 chemicals, including assay data for three chemicals reported previously by the authors. As a result, the improved transformation assay showed a concordance of 73.5% with a rodent bioassay, a sensitivity for carcinogens of 71.4%, and a specificity for detection of noncarcinogens of 76.9%. The improved transformation assay detected all of the genotoxic carcinogens, and five of 11 nongenotoxic carcinogens as positive. It can be expected that the improved transformation assay will be able to detect not only genotoxic carcinogens with high probability but also approximately 50% of nongenotoxic carcinogens within about 3 weeks. Hence, these preliminary findings suggest that our improved transformation assay will be a reliable and useful short-term test procedure of screening for potential carcinogens, and it encourages us to conduct further experiments on many carcinogens and noncarcinogens. PMID- 12849734 TI - Ocular toxicity of some corneal penetration enhancers evaluated by electrophysiology measurements on isolated rabbit corneas. AB - The influence on electrical resistance and membrane potential of rabbit corneas in vitro of some chemicals used as adjuvants in ophthalmic formulations was investigated, in the attempt to correlate changes in electrophysiological properties of the corneal tissue (possibly indicative of toxic/damaging effects to the corneal epithelium), with the promoting effect of the substances on transcorneal permeation in vitro of timolol maleate (TM). The chemicals, tested at different concentrations, were benzalkonium chloride (BAC), sodium ethylenediaminetetraacetate (EDTA), polyoxyethylene-20-stearyl ether (PSE), polyethoxylated castor oil (PCO), deoxycholic acid sodium salt (DC) and cetylpyridinium chloride (CPC). For these substances, definite correlations were found between promoting activity for permeation of TM and modification of electrophysiological parameters. These parameters were in all cases significantly altered by all agents at all concentrations after a 5-h contact. However, after a 1-h contact, 0.001% PSE and CPC did not significantly modify the corneal resistance, while PCO and PSE did not significantly modify the transcorneal potential at the tested concentrations. Only 0.001% PSE, a nonionic surfactant used as solubilizer and emulsifier, active as promoter for TM, did not modify both electrophysiological parameters to a significant extent after 1 h. The results of this study indicate correlations between ocular toxicity, promoting activity for transcorneal permeation of timolol and modification of the electrophysiological parameters. PMID- 12849735 TI - Hzf-3 expression in the amygdala after establishment of conditioned taste aversion. AB - We studied the regulation of the expression of the inducible orphan nuclear receptor known as HZF-3 (or Nurr1) in acquisition of conditioned taste aversion in rats. Our results show that HZF-3 expression in the lateral/basolateral (LA/BLA) amygdala complex was significantly up-regulated when both the conditioned and the unconditioned stimuli were paired, but not when either of the stimuli was presented alone. Induction in the LA/BLA had a faster onset than induction in the central nucleus of the amygdala. The results implicate HZF-3 in the acquisition of associative aversive experiences. PMID- 12849736 TI - Ultrastructural study of gap junctions between dendrites of parvalbumin containing GABAergic neurons in various neocortical areas of the adult rat. AB - Parvalbumin (PV)-containing GABAergic neurons in the hippocampus form dual networks linked by both dendrodendritic gap junctions and mutual inhibitory synapses. Recent physiological studies have demonstrated similar functional connectivity among cortical GABAergic neurons, but the corresponding structures have not been fully analyzed at the electron microscopic level. In this study we examined detailed ultrastructural features of gap junctions between PV neurons in the mature neocortex. Light microscopic observations and confocal laser scanning microscopy revealed frequent dendrodendritic contacts between PV neurons. Electron microscopic analysis provided direct morphological evidence for the existence of gap junctions between 22 pairs of PV-immunoreactive dendrites in the visual, auditory, and somatosensory cortices. Their ultrastructural features that were characteristic of immunolabeled profiles were consistent with the general structure of gap junctions. In one case a gap junction coexisted with a dendrodendritic chemical synapse, making a mixed synapse. Importantly, we also encountered a gap junction between PV positive and negative, presumptive non principal cell-derived, dendrites. Quantitative analysis was made in 16 pairs of PV positive dendrites forming gap junctions in the infragranular layers of the somatosensory cortex. Diameters of these dendrites ranged from 0.3 to 2.7 microm, suggesting diverse locations of gap junctions along the proximal-distal axis of dendritic trees, but the majority (81%) were less than 1 microm. The mean size of gap junctions along apposing membranes was 0.22+/-0.09 microm. By using this size, the theoretical value of a junctional conductance was estimated to be 2.1 5.3 nS. Dendrites of PV neurons in the infragranular layers of the somatosensory cortex were reconstructed light microscopically and the sites of contacts with other PV neurons were mapped. Although these contacts do not necessarily imply gap junctional coupling, their number (5.3+/-2.3 per cell, n=11) suggested the degree of connectivity of less than 10 coupling from single PV neurons with others. Sholl analysis revealed that only 38% of their dendrites occurred within 200 microm from the soma. The present study demonstrated detailed ultrastructural features of gap junctions between mature cortical PV neurons. These features will facilitate not only identification of gap junctions in variously labeled neurons but also analysis of their functional aspects by enabling theoretical estimate of their junctional conductances. PMID- 12849737 TI - Quinacrine attenuates increases in divalent metal transporter-1 and iron levels in the rat hippocampus, after kainate-induced neuronal injury. AB - The present investigation was carried out to elucidate the effect of the antimalarial drug quinacrine on levels of expression of the non-heme iron transporter, divalent metal transporter-1 (DMT1) and iron, in the hippocampus of rats after kainate treatment. The untreated hippocampus was lightly stained for DMT1, while an increase in DMT1 staining in astrocytes in the degenerating cornu ammonis (CA) fields, after kainate lesions. The increased DMT1 immunoreactivity was correlated with increased levels of Fe3+ and Fe2+ staining in the CA fields, as demonstrated by iron histochemistry (Perl's and Turnbull's blue stain for Fe3+ and Fe2+). The increases in DMT1 and iron staining were significantly attenuated by quinacrine. Rats injected with kainate and daily i.p. injections of quinacrine (5 mg/kg) for 7 days or 2 weeks showed significantly lower levels of DMT1 immunoreactivity and iron staining, compared with rats injected with kainate and saline. These results show that DMT1 expression is closely linked to iron levels, and provide further support for a crucial role that DMT1 plays in iron accumulation in the degenerating hippocampus. PMID- 12849738 TI - Concomitant up-regulation of astroglial high and low affinity nerve growth factor receptors in the CA1 hippocampal area following global transient cerebral ischemia in rat. AB - We have examined the effect of global transient cerebral ischemia, evoked in rat by 10 min of cardiac arrest, upon the changes in the cellular expression of two nerve growth factor (NGF) receptors (TrkA and p75) in the hippocampus. We have used immunocytochemical procedures, including a quantitative analysis of staining, along with some quantitative morphological analyses. We have found, under ischemic conditions, a decrease of TrkA immunoreactivity in degenerating CA1 pyramidal neurons and in neuropil. On the other hand, a strong, ischemia induced up-regulation of TrkA and p75 immunoreactivity was observed in the majority of reactive astroglia population in the adjacent CA1 hippocampal region. The colocalization of the two receptors in the same reactive astroglial cells was evidenced by double immunostaining and further supported by quantitative morphological analysis of TrkA and p75 immunoreactive glial cells. Our data implicate the involvement of NGF receptors in the postischemic regulation of astrocytic function; however, the lack of NGF receptor expression on some astrocytes suggests heterogeneity of astroglia population. Our results also indicate that the lack of neuroprotective action of astroglial NGF induced in the ischemic hippocampus [J Neurosci Res 41 (1995) 684; Acta Neurobiol Exp 57 (1997) 31; Neuroscience 91 (1999) 1027] is not caused by a paucity of NGF receptors but may rather be due to the counteraction of some proinflammatory substances, released simultaneously by glia cells. On the other hand, the up-regulated astroglial TrkA receptor may be an important target for exogenous NGF, which, as previously described [J Neurosci 11 (1991) 2914; Neurosci Lett 141 (1992) 161], exerts a neuroprotective effect in ischemia. PMID- 12849739 TI - Inhibiting cell proliferation during formation of the glial scar: effects on axon regeneration in the CNS. AB - Following a CNS lesion many glial cell types proliferate and/or migrate to the lesion site, forming the glial scar. The majority of these cells express chondroitin sulphate proteoglycans (CS-PGs), previously shown to inhibit axonal growth. In this study, in an attempt to diminish glial scar formation and improve axonal regeneration, proliferating cells were eliminated from the lesion site. Adult rats received a continuous infusion of 2% cytosine-D-arabinofuranoside (araC) or saline for 7 days over the lesion site, immediately following a unilateral transection of the right medial forebrain bundle. Additional groups of rats that received subdural infusions prior to the lesion, and lesioned rats which received no infusion, were also compared in the analyses. Animals were killed at 4, 7, 12 or 18 days post-lesion (dpl) and immunohistochemistry was used to determine the effects of these treatments on tyrosine hydroxylase (TH) lesioned axons, and on the injury response of glial cells. Almost complete elimination of NG2 oligodendrocyte progenitor cells from the lesion site was seen up to 7 dpl in araC-infused animals; reduced numbers of reactive CD11b microglia were also seen but no effects were seen on the injury response of GFAP astrocytes. Significantly more TH axons were seen distal to the lesion in araC treated brains, but these numbers dwindled by 18 dpl. PMID- 12849740 TI - Pb2+ inhibits the hyperpolarization-activated current in acutely isolated dorsal root ganglion neurons. AB - The hyperpolarization-activated h channel current (Ih) reported to be present in acutely isolated rat dorsal root ganglion (DRG) neurons is inhibited by Cs+ and ZD7288. It was recently reported that lead (Pb2+) inhibits voltage-gated Ca2+ and K+ channels in DRG neurons but the effect of Pb2+ on Ih has so far not been reported. Using whole-cell patch clamp technique we show that Pb2+ specifically inhibited Ih. External application of 0.1, 1 and 10 microM Pb2+ reversibly reduced the amplitude of Ih in a dose-dependent manner, with an IC50 value of 3.7 microM and a Hill coefficient of 1.1. Pb2+ shifted the activation curve of Ih by 9.3 mV but had no effect on the slope factor. Pb2+ inhibited Ih in a voltage dependent manner and slowed down the activation process, indicating an action of Pb2+ on the activation kinetics of h channels. Our studies thus demonstrated that Pb2+ is a dose-dependent, voltage-dependent and reversible blocker of Ih in DRG neurons. PMID- 12849741 TI - Characterization of an apamin-sensitive potassium current in suprachiasmatic nucleus neurons. AB - In neurons of the suprachiasmatic nucleus, spike frequency adaptation and membrane afterhyperpolarization occur during a train of action potentials. Extracellular Ca2+ may regulate neuronal excitability by several mechanisms, including activation of small conductance and large conductance Ca(2+)-activated K+ channels. The overall goal of this study was to examine the role of Ca(2+) activated K+ currents in individual suprachiasmatic nucleus neurons. To this end, we used the nystatin-perforated patch technique to record currents from suprachiasmatic nucleus neurons. Iberiotoxin and tetraethylammonium, antagonists of large conductance Ca(2+)-activated K+ channels, had no effect on the membrane afterhyperpolarization. However, antagonists of small conductance Ca(2+) activated K+ channels, apamin and d-tubocurarine, reduced the amplitude of the membrane afterhyperpolarization and inhibited the spike frequency adaptation that occurred during a train of action potentials. Although there was no significant difference in membrane AHP between different portions of the circadian day, apamin and d-tubocurarine increased the spontaneous firing frequency of suprachiasmatic nucleus neurons during the daytime. In voltage-clamp mode, membrane depolarization-activated currents were followed by an outward tail current reversing near the K+ equilibrium potential. The tail current decayed with a time constant of 220 ms at +20 mV and 149 ms at -40 mV. Apamin irreversibly and d-tubocurarine reversibly inhibited the tail current. The tail current amplitude was also reduced by the GABAA receptor antagonist, bicuculline methiodide, while picrotoxin (another GABAA receptor antagonist) was without effect. Removal of extracellular Ca2+ or the addition of Cd2+ reversibly inhibited the tail current. These results indicate that apamin- and d tubocurarine-sensitive small conductance Ca(2+)-activated K+ channels have a modulatory function on the action potential firing frequency as well as the membrane afterhyperpolarization that follows a train of action potentials in suprachiasmatic nucleus neurons. Importantly, our data also indicate that a portion of the effects of bicuculline methiodide on suprachiasmatic nucleus neurons may be mediated by inhibition of small conductance Ca(2+)-activated K+ channels. PMID- 12849742 TI - Involvement of amygdala networks in epileptiform synchronization in vitro. AB - We used field potential and intracellular recordings in rat brain slices that included the hippocampus, a portion of the basolateral/lateral nuclei of the amygdala (BLA) and the entorhinal cortex (EC). Bath application of the convulsant 4-aminopyridine (50 microM) to slices (n=12) with reciprocally connected areas, induced short-lasting interictal-like epileptiform discharges that (i) occurred at intervals of 1.2-2.8 s, (ii) originated in CA3, and (iii) spread to EC and BLA. Cutting the Schaffer collaterals abolished them in both parahippocampal areas where slower interictal-like (interval of occurrence=4-17 s) and prolonged ictal-like discharges (duration=15+/-6.9 s, mean+/-S.D., n=7) appeared. These new types of epileptiform activity originated in either EC or BLA. Similar findings were obtained in slices (n=19) in which the hippocampus outputs were not connected with the EC and BLA under control conditions. Cutting the EC-BLA connections made independent slow interictal- and ictal-like activities appear in both areas (n=5). NMDA receptor antagonism (n=6) abolished ictal-like discharges and reduced the duration of the slow interictal-like events. Repetitive stimulation of BLA at 0.5-1 Hz in Schaffer collateral cut slices, induced interictal-like epileptiform depolarizations in EC and reversibly blocked ictal like activity (n=14). Thus, CA3 outputs in intact slices entrain EC and BLA networks into an interictal-like pattern that inhibits the propensity of these parahippocampal areas to generate prolonged ictal-like paroxysms. Accordingly, NMDA receptor-dependent ictal-like events are initiated in BLA or EC once the propagation of CA3-driven interictal-like discharges to these areas is abated by cutting the Schaffer collaterals. Similar inhibitory effects also occur by activating BLA outputs directed to EC at rates that mimic the CA3-driven interictal-like pattern. PMID- 12849743 TI - Up-regulation of P2X2, P2X4 receptor and ischemic cell death: prevention by P2 antagonists. AB - In the present work we examined the involvement of selected P2X receptors for extracellular ATP in the onset of neuronal cell death caused by glucose/oxygen deprivation. The in vitro studies of organotypic cultures from hippocampus evidenced that P2X2 and P2X4 were up-regulated by glucose/oxygen deprivation. Moreover, we showed that ischemic conditions induced specific neuronal loss not only in hippocampal, but also in cortical and striatal organotypic cultures and the P2 receptor antagonists basilen blue and suramin prevented these detrimental effects. In the in vivo experiments we confirmed the induction of P2X receptors in the hippocampus of gerbils subjected to bilateral common carotid occlusion. In particular, P2X2 and P2X4 proteins became significantly up-regulated, although to different extent and in different cellular phenotypes. The induction was confined to the pyramidal cell layer of the CA1 subfield and to the transition zone of the CA2 subfield and it was coincident with the area of neuronal damage. P2X2 was expressed in neuronal cell bodies and fibers in the CA1 pyramidal cell layer and in the strata oriens and radiatum. Intense P2X4 immunofluorescence was localized to microglia cells. Our results indicate a direct involvement of P2X receptors in the mechanisms sustaining cell death evoked by metabolism impairment and suggest the use of selected P2 antagonists as effective neuroprotecting agents. PMID- 12849744 TI - Ciliary neurotrophic factor in the olfactory bulb of rats and mice. AB - Ciliary neurotrophic factor (CNTF) is primarily regarded as an astrocytic lesion factor, promoting neuronal survival and influencing plasticity processes in deafferented areas of the CNS. Postnatal loss of neurons in CNTF-deficient mice indicates a function of the factor also under physiological conditions. In the olfactory bulb, where neurogenesis, axo- and synaptogenesis continue throughout life, CNTF content is constitutively high. The cellular localization of CNTF in the rat olfactory bulb is not fully resolved, and species differences between mouse and rat are not yet characterized. In the present study, four different CNTF antibodies and double immunolabeling with specific markers for glial and neuronal cells were used to study the cellular localization of CNTF in rat and mouse olfactory bulb. Specificity of the detection was checked with tissue from CNTF-deficient mice, and investigations were complemented by immunolocalization of reporter protein in mice synthesizing beta-galactosidase under control of the CNTF promoter (CNTF lacZ-knock-in mice). In both species, CNTF localized to ensheathing cell nuclei, cell bodies and axon-enveloping processes. Additionally, individual axons of olfactory neurons were CNTF immunoreactive. Both CNTF protein content and immunoreaction intensity were lower in mice than in rats. Scattered lightly CNTF-reactive cells were found in the granular and external plexiform layers in rats. Some CNTF-positive cells were associated with immunoreactivity for the polysialylated form of the neural cell adhesion molecule, which is expressed by maturing interneurons derived from the rostral migratory stream. In CNTF lacZ-knock-in mice, beta-galactosidase reactivity was found in ensheathing cells of the olfactory nerve layer, and in cells of the glomerular, external plexiform and granular layers. The study proves that CNTF is localized in glial and neuronal structures in the rodent olfactory bulb. Results in mice provide a basis for investigations concerning the effects of a lack of the factor in CNTF deficient mice. PMID- 12849745 TI - Hyperbaric oxygen therapy protects against mitochondrial dysfunction and delays onset of motor neuron disease in Wobbler mice. AB - The Wobbler mouse is a model of human motor neuron disease. Recently we reported the impairment of mitochondrial complex IV in Wobbler mouse CNS, including motor cortex and spinal cord. The present study was designed to test the effect of hyperbaric oxygen therapy (HBOT) on (1) mitochondrial functions in young Wobbler mice, and (2) the onset and progression of the disease with aging. HBOT was carried out at 2 atmospheres absolute (2 ATA) oxygen for 1 h/day for 30 days. Control groups consisted of both untreated Wobbler mice and non-diseased Wobbler mice. The rate of respiration for complex IV in mitochondria isolated from motor cortex was improved by 40% (P<0.05) after HBOT. The onset and progression of the disease in the Wobbler mice was studied using litters of pups from proven heterozygous breeding pairs, which were treated from birth with 2 ATA HBOT for 1 h/day 6 days a week for the animals' lifetime. A "blinded" observer examined the onset and progression of the Wobbler phenotype, including walking capabilities ranging from normal walking to jaw walking (unable to use forepaws), and the paw condition (from normal to curled wrists and forelimb fixed to the chest). These data indicate that the onset of disease in untreated Wobbler mice averaged 36+/ 4.3 days in terms of walking and 40+/-5.7 days in terms of paw condition. HBOT significantly delayed (P<0.001 for both paw condition and walking) the onset of disease to 59+/-8.2 days (in terms of walking) and 63+/-7.6 days (in terms of paw condition). Our data suggest that HBOT significantly ameliorates mitochondrial dysfunction in the motor cortex and spinal cord and greatly delays the onset of the disease in an animal model of motor neuron disease. PMID- 12849746 TI - Normal retinal development and retinofugal projections in mice lacking the retina specific variant of actin-binding LIM domain protein. AB - The actin-binding LIM domain protein (abLIM) is the mammalian homologue of UNC 115, a protein mediating axon guidance in C. elegans. AbLIM is widely expressed with three isoforms differing from one another by the length of their amino termini. Experiments utilizing dominant-negative mutants in the chick retina suggested a role for abLIM in axon path finding in retinal ganglion cells (RGCs). To investigate which variant is involved in the regulation of mammalian RGC axon guidance, we analyzed their expression profile in mice. The longest variant, abLIM-L, is highly enriched in the ganglion cell layer. AbLIM-L is up-regulated postnatally which temporally overlaps with the period of RGC axon remodeling. In contrast, the abLIM-M and abLIM-S variants are widespread and remain relatively constant through development. By selective gene targeting, we ablated abLIM-L to explore its functional significance in vivo. AbLIM-L mutant mice exhibit no apparent morphological or functional defects in photoreceptors and inner retinal neurons. Retinofugal projections and synaptic maturation also appear normal. These data suggest that abLIM-M is likely the isoform performing the essential function related to axon guidance. PMID- 12849747 TI - Retinotopic map plasticity in adult cat visual cortex is accompanied by changes in Ca2+/calmodulin-dependent protein kinase II alpha autophosphorylation. AB - In adult cats, the induction of homonymous binocular central retinal lesions causes a dramatic reorganization of the topographic map in the sensory-deprived region of the primary visual cortex. To investigate the possible involvement of the alpha-subunit of the calcium/calmodulin dependent protein kinase type II (alphaCaMKII) in this form of brain plasticity, we performed in situ hybridization and Western blotting experiments to analyze mRNA, protein and autophosphorylation levels of this multifunctional kinase. No differences in the mRNA or protein levels were observed between the central, sensory-deprived and the peripheral, non-deprived regions of area 17 of retinal lesion animals or between corresponding cortical regions of normal control animals. Western blotting with an alphaCaMKII threonine-286 phosphorylation-state specific antiserum consistently showed a small, albeit not significant, increase of alphaCaMKII autophosphorylation in the central versus the peripheral region of cortical area 17, and this both in normal subjects as well as in retinal lesion animals with a 3-day post-lesion survival time. In contrast, a post-lesion survival time of 14 days resulted in a alphaCaMKII autophosphorylation level that was four times higher in visually-deprived area 17 than in the non-deprived cortical region. This increased phosphorylation state is not a direct consequence of the decrease in visual activity in these neurons, because we would have expected to see a similar change at shorter or longer post-lesion survival times or in the visually deprived visual cortex of animals in which the left optic tract and the corpus callosum were surgically cut. No such changes were observed, leading to the conclusion that the phosphorylation changes observed at 14 days are related to a delayed reorganization of the retinotopic map of the striate cortex. PMID- 12849748 TI - X11alpha modulates secretory and endocytic trafficking and metabolism of amyloid precursor protein: mutational analysis of the YENPTY sequence. AB - The neuronal adaptor X11alpha interacts with the conserved -GYENPTY- sequence in the C-terminus of amyloid precursor protein (APP) or its Swedish mutation (APPswe) to inhibit Abeta40 and Abeta42 secretion. We hypothesized that the -YENP motif essential for APP endocytosis is also essential for X11alpha-mediated effects on APP trafficking and metabolism, and that X11alpha modulates APP metabolism in both secretory and endocytic pathways. X11alpha failed to interact with the endocytic-defective APPswe mutants Y738A, N740A, or P741A, and thus did not modulate their trafficking or metabolism. However, endocytic-competent APPswe Y743A had unique trafficking and metabolism including a prolonged half-life and increased secretion of catabolites compared with APPswe. In contrast to endocytic defective mutants, X11alpha interacted with APPswe Y743A as well as with APPswe. Thus, similar to APPswe, coexpression of X11alpha with APPswe Y743A retarded its maturation, prolonged its half-life, and inhibited APPs, Abeta40, and Abeta42 secretion. Collectively, these data suggest that by direct interaction with the APPswe -YENP- motif in the cytoplasmic tail, X11alpha modulated its trafficking and processing in both secretory and endocytic compartments, and may reduce secretion of Abeta generated in either pathway. PMID- 12849749 TI - The neuronal distribution of cannabinoid receptor type 1 in the trigeminal ganglion of the rat. AB - Cannabinoid compounds have been shown to produce antinociception and antihyperalgesia by acting upon cannabinoid receptors located in both the CNS and the periphery. A potential mechanism by which cannabinoids could inhibit nociception in the periphery is the activation of cannabinoid receptors located on one or more classes of primary nociceptive neurons. To address this hypothesis, we evaluated the neuronal distribution of cannabinoid receptor type 1 (CB1) in the trigeminal ganglion (TG) of the adult rat through combined in situ hybridization (ISH) and immunohistochemistry (IHC). CB1 receptor mRNA was localized mainly to medium and large diameter neurons of the maxillary and mandibular branches of the TG. Consistent with this distribution, in a de facto nociceptive sensory neuron population that exhibited vanilloid receptor type 1 immunoreactivity, colocalization with CB1 mRNA was also sparse (<5%). Furthermore, very few neurons (approximately 5%) in the peptidergic (defined as calcitonin gene-related peptide- or substance P-immunoreactive) or the isolectin B4-binding sensory neuron populations contained CB1 mRNA. In contrast, and consistent with the neuron-size distribution for CB1, nearly 75% of CB1-positive neurons exhibited N52-immunoreactivity, a marker of myelinated axons. These results indicate that in the rat TG, CB1 receptors are expressed predominantly in neurons that are not thought to subserve nociceptive neurotransmission in the noninjured animal. Taken together with the absence of an above background in situ signal for CB2 mRNA in TG neurons, these findings suggest that the peripherally mediated antinociceptive effects of cannabinoids may involve either as yet unidentified receptors or interaction with afferent neuron populations that normally subserve non-nociceptive functions. PMID- 12849750 TI - Ganglioside/calmodulin kinase II signal inducing cdc42-mediated neuronal actin reorganization. AB - Cell surface glycoconjugates are thought to mediate cell-cell recognition and play roles in neuronal development and functions. We demonstrated here that exposure of neuronal cells to nanomolar levels of gangliosides Neu5Acalpha 8Neu5Acalpha 3Galbeta 4GlcCer, Galbeta 3GalNAcbeta 4(Neu5Acalpha 8Neu5Acalpha 3)Galbeta 4GlcCer (GD1b), Neu5Acalpha 3Galbeta 3GalNAcbeta 4(Neu5Acalpha 8Neu5Acalpha 3)Galbeta 4GlcCer (GT1b) or its oligosaccharide portion induced a rapid and transient activation of Ca2+/calmodulin-dependent protein kinase II (CaM-KII) in the subplasmalemma. Galbeta 3GalNAcbeta 4(Neu5Acalpha 3)Galbeta 4GlcCer (GM1), GalNAcbeta 4(Neu5Acalpha 3)Galbeta 4GlcCer, Neu5Acalpha 3Galbeta 4GlcCer, Neu5Acalpha 3Galbeta 3GalNAcbeta 4(Neu5Acalpha 3)Galbeta 4GlcCer (GD1a), and Neu5Acalpha 8Neu5Acalpha 3Galbeta 3GalNAcbeta 4(Neu5Acalpha 8Neu5Acalpha 3) Galbeta 4GlcCer were ineffective. GT1b and GD1b stimulated transient elevation of bulk cytosolic Ca2+ levels while GM1 slightly elevated the levels and GD1a did not. Thus, the cytosolic Ca2+ elevation by the gangliosides may trigger the CaM KII activation. The treatment was accompanied by peripheral actin polymerization and filopodia formation in NG108-15 cells and primary hippocampal neurons, but not in glial cells. CaM-KII inhibitors blocked both CaM-KII activation and the subsequent filopodia formation. A small G-protein cdc42 was a potential downstream target of CaM-KII activated by the gangliosides. These results suggest that oligosaccharides of the gangliosides serve as potential regulators of the filopodia formation in neuronal cells by triggering the activation of CaM-KII followed by cdc42 up-regulation via a cell surface receptor-like component. The filopodia formation induced by the gangliosides may have a physiological relevance because long-term exposure of hippocampal neurons to GT1b oligosaccharide induced advanced dendritogenesis. Furthermore, exposure of cerebellar neurons to GT1b oligosaccharide facilitated CaM-KII-dependent dendritic outgrowth and branch formation of cerebellar Purkinje neurons, in which actin isoforms were localized to motile structures in dendrites. Thus, the ganglioside/CaM-KII signal plays a role in modulating dendritic morphogenesis by inducing cdc42-mediated actin reorganization. PMID- 12849751 TI - Hypertension-induced changes in monoamine receptors in the prefrontal cortex of rhesus monkeys. AB - Hypertension affects approximately 60 million people in the United States. Recent studies have demonstrated that hypertension may produce progressive changes in the CNS. The present study is focused on reports in the literature that hypertension may significantly alter neurotransmitter systems, particularly dopamine (DA) and norepinephrine (NE). To address this, DA and norepinephrine (NE) receptor binding was assessed in the prefrontal cortex (PFC) of 15 male rhesus monkeys using on-the-slide in vitro assays for the DA1, NE alpha1 and NE alpha2 receptors as well as for the DA and NE uptake transporters. Eight monkeys underwent surgical coarctation of the mid-thoracic aorta which produced sustained, untreated hypertension as defined by a systolic pressure above 150 mm Hg. Compared with normotensive controls, chronic, untreated hypertension produced a significant decrease in DA1 and NE alpha1 receptor binding and an increase in DA uptake (DAU) receptor binding in the prefrontal cortex. While the mechanisms by which untreated hypertension alters DA and NE receptors is not known, the use of this non-human primate model should provide the means to uncover neurobiological changes that occur with untreated hypertension. PMID- 12849752 TI - Pifithrin-alpha suppresses p53 and protects cochlear and vestibular hair cells from cisplatin-induced apoptosis. AB - Cisplatin, a commonly used antineoplastic agent, destroys the sensory hair cells in the cochlear and vestibular system leading to irreversible hearing loss and balance problems. Cisplatin-induced hair cell damage presumably occurs by apoptosis. Recent studies suggest that p53 may play an important role initiating cisplatin-induced apoptosis in some cell types. To determine if p53 plays a role in cisplatin-mediated hair cell loss, cochlear and utricular organotypic cultures were prepared from postnatal day 3-4 rats and treated with cisplatin or cisplatin plus pifithrin-alpha (PFT), a p53 inhibitor. Control cultures were devoid of p53 immunolabeling, caspase-1 and caspase-3 labeling and p53 protein was absent from Western blots. Cisplatin (1-10 microg/ml) caused a dose-dependent loss of hair cells in cochlear and utricular cultures, up-regulated phospho-p53 serine 15 immunolabeling, increased the expression of phospho-p53 serine 15 in Western blots from 6 to 48 h after the onset of cisplatin-treatment, and increased caspase-1 and caspase-3 labeling in cochlear and vestibular cultures. Addition of PFT (20-100 microM) to cisplatin-treated cochlear and utricular cultures resulted in a dose-dependent increase in hair cell survival; suppressed the expression of p53 in Western blots and eliminated caspase-1 and caspase-3 labeling in cultures. These results suggest that the tumor suppressor protein, p53, plays a critical role in initiating apoptosis in cochlear and vestibular hair cells. Temporary suppression of p53 with PFT provides significant protection against cisplatin induced hair cell loss and offers the potential for reducing the ototoxic, vestibulotoxic and neurotoxic side effects of cisplatin. PMID- 12849753 TI - Light microscopic study of GluR1 and calbindin expression in interneurons of neocortical microgyral malformations. AB - Rat neocortex that has been injured on the first or second postnatal day (P0-1) develops an epileptogenic, aberrantly layered malformation called a microgyrus. To investigate the effects of this developmental plasticity on inhibitory interneurons, we studied a sub-population of GABAergic cells that co-express the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor GluR1 subunit and the calcium-binding protein, calbindin (CB). Both malformed and control cortex of adult (P40-60) animals contained numerous interneurons double-stained for CB and GluR1. Immunoreactivity (IR) for CB was up-regulated in perikarya of interneurons within supragranular layers of control cortex between P12 and P40. However, in malformed adult (P40) cortex, CB-IR levels were significantly lower than in adult controls, and fell midway between levels in immature and adult control animals. Between P12 and P40, GluR1-IR was down-regulated in perikarya of interneurons in control cortex. Somatic GluR1-IR levels in malformed adult (P40) cortex were not different from adult controls. These neurons formed a dense plexus of highly GluR1-positive spiny dendrites within layer II. The dendritic plexus in the malformation was more intensely GluR1-immunoreactive than that in layer II of control cortex. This was due to apparent changes in thickness and length of dendrites, rather than to significant changes in the number of interneuronal perikarya in the microgyral cortex. Results indicate that the population of GluR1/CB-containing interneurons is spared in malformed microgyral cortex, but that these cells sustain lasting decreases in their somatic expression of calbindin and alterations of dendritic structure. Potential functional implications of these findings are discussed. PMID- 12849754 TI - Transient attenuation of protein kinase Cepsilon can terminate a chronic hyperalgesic state in the rat. AB - Recently we demonstrated that a single 3-day episode of carrageenan-induced acute cutaneous inflammation can create a chronic state of increased susceptibility to inflammatory hyperalgesia. In this latent "primed" state, although there is no ongoing hyperalgesia, the hyperalgesic response to subsequent challenges with inflammatory agent (prostaglandin E2; PGE2) is greatly enhanced. Furthermore, the PGE2-induced hyperalgesia in primed skin was found to require activity of the epsilon isozyme of protein kinase C (PKCepsilon), a second messenger that is not required for PGE2-induced hyperalgesia in control animals. In the present study we tested the hypothesis that activity of PKCepsilon not only plays a critical role in the expression of primed PGE2-induced hyperalgesia, but also in the development and maintenance of the primed state itself. Antisense oligodeoxynucleotide was employed to produce a decrease in PKCepsilon in the nerve, verified by Western blot analysis. PKCepsilon was found to be essential both for the development of carrageenan-induced hyperalgesic priming, as well as for the maintenance of the primed state. Furthermore, hyperalgesic priming could be induced by an agonist of PKCepsilon (pseudo-receptor octapeptide for activated PKCepsilon) at a dose that itself causes no hyperalgesia. The finding that transient inhibition of PKCepsilon can not only prevent the development of priming, but can also terminate a fully developed state of priming suggests the possibility that selective targeting PKCepsilon might be an effective new strategy in the treatment of chronic inflammatory pain. PMID- 12849755 TI - Interleukin-1 beta in the hypothalamus potentiates feline defensive rage: role of serotonin-2 receptors. AB - The neurochemistry of aggression and rage has largely focused on the roles played by neurotransmitters and their receptor mechanisms. In contrast, little attention has been given to the possible functions of other substances. Interleukin-1beta is an immune and brain-derived cytokine that is present in the hypothalamus. Functionally, interleukin-1 has been shown to induce the release of serotonin (5 HT), a neurotransmitter known to potently affect aggression and rage behavior. Thus, the goal of the present study was to test the hypothesis that interleukin 1beta in the medial hypothalamus could modulate defensive rage behavior in the cat. In the first experiment, electrical stimulation of sites in the medial hypothalamus from which defensive rage could be elicited and where microinjections of specific compounds were later placed, facilitated defensive rage elicited from the periaqueductal gray (PAG), thus demonstrating the functional relationship between these two regions. In the second experiment, microinjections of relatively low doses of interleukin-1beta into the medial hypothalamus potentiated defensive rage behavior elicited from the midbrain periaqueductal gray in a dose-related manner. In the third experiment, pretreatment with a selective 5-HT2 receptor antagonist, LY-53857, blocked the facilitating effects of interleukin-1beta upon defensive rage. These findings reveal for the first time that brain cytokines can dramatically alter aggressive behavior. In particular, interleukin-1beta in the medial hypothalamus potentiates defensive rage behavior elicited from the periaqueductal gray in the cat, and the potentiating effects of interleukin-1beta on this form of emotional behavior are mediated via a 5-HT2 receptor mechanism. PMID- 12849756 TI - A comparative magnetoencephalographic study of cortical activations evoked by noxious and innocuous somatosensory stimulations. AB - We recorded somatosensory-evoked magnetic fields and potentials produced by painful intra-epidermal stimulation (ES) and non-painful transcutaneous electrical stimulation (TS) applied to the left hand in 12 healthy volunteers to compare cortical responses to noxious and innocuous somatosensory stimulations. Our results revealed that cortical processing following noxious and innocuous stimulations was strikingly similar except that the former was delayed approximately 60 ms relative to the latter, which was well explained by a difference in peripheral conduction velocity mediating noxious (Adelta fiber) and innocuous (Abeta fiber) inputs. The first cortical activity evoked by both ES and TS was in the primary somatosensory cortex (SI) in the hemisphere contralateral to the stimulated side. The following activities were in the bilateral secondary somatosensory cortex (SII), insular cortex, cingulate cortex, anterior medial temporal area and ipsilateral SI. The source locations did not differ between the two stimulus modalities except that the dipole for insular activity following ES was located more anterior to that following TS. Both ES and TS evoked vertex potentials consisting of a negativity followed by a positivity at a latency of 202 and 304 ms, and 134 and 243 ms, respectively. The time course of the vertex potential corresponded to that of the activity of the medial temporal area. Our results suggested that cortical processing was similar between noxious and innocuous stimulation in SI and SII, but different in insular cortex. Our data also implied that activities in the amygdala/hippocampal formation represented common effects of noxious and tactile stimulations. PMID- 12849757 TI - Selective age-related loss of calbindin-D28k from basal forebrain cholinergic neurons in the common marmoset (Callithrix jacchus). AB - A significant number of the cholinergic neurons in the basal forebrain of the primate, but not the rodent brain contain the calcium binding protein calbindin D28k (CB). Previous experiments in our laboratory have demonstrated a substantial age-related loss of CB from the human basal forebrain cholinergic neurons (BFCN). The present study investigated the possible age-related loss of CB from the BFCN in a non-human primate species, the common marmoset (Callithrix jacchus). Quantitative analysis of matching sections as well as unbiased stereological determination of neuronal number were used in 16 adult marmosets ranging in age between 2 and 15 years. No significant changes were observed in the number of choline acetyltransferase-positive BFCN when a group of young animals (< or =4 years) was compared with a 6-8-year-old group and a 9-15-year-old group. Similarly, no age-related changes were observed in Nissl-stained magnocellular basal forebrain (putatively cholinergic) neurons. In contrast, the BFCN of the two older groups of animals displayed a significant loss of CB. The age-related loss of CB occurred in all sectors of the BFCN, but was greatest in the anterior sector of this cell group. The CB loss was neurochemically specific since the BFCN in the older groups of animals continued to express other markers such as high and low affinity neurotrophin receptors. The age-related loss of CB from the marmoset BFCN was also regionally selective as CB positive neurons in other structures, such as the cerebral cortex and the striatum displayed no apparent age-related changes. These results indicate that the marmoset BFCN display a significant and selective age-related loss of CB reminiscent of that observed in the human. Therefore, the common marmoset represents an appropriate animal model in which the consequences of BFCN CB loss can be investigated in depth. Loss of CB from the aged BFCN is likely to reduce the capacity of these neurons to buffer intracellular calcium and to leave them vulnerable to insults which can result in increased calcium levels. The vulnerability of the CB-negative BFCN in the aged marmoset to various insults which disturb calcium homeostasis remains to be investigated. PMID- 12849758 TI - The involvement of dopamine and nitric oxide in the endocrine and behavioural action of endomorphin-1. AB - Previous publications have demonstrated a prominent central and corticotropin releasing hormone-mediated action of the endomorphins (EMs) on both open-field behaviour and the hypothalamo-pituitary-adrenal (HPA) axis. In the present experiments, the direct action of endomorphin-1 (EM1) on pituitary adrenocorticotropic hormone (ACTH) release, adrenal corticosterone secretion and the roles of nitric oxide (NO) and dopamine (DA) in the HPA and behavioural responses elicited by EM1 were investigated in mice. In vitro perifusion studies indicated that the action of EM1 on the HPA system appears to be confined to the hypothalamus, as EM1 did not influence the corticosterone secretion from adrenal slices and moderately attenuated the ACTH release from anterior pituitary slices. In in vivo experiments, NG-nitro-L-arginine (L-NNArg) pretreatment brought about a profound inhibition of both the endocrine and the behavioural responses. On the other hand, haloperidol completely abolished the increases in square crossing and rearing, without affecting corticosterone release. The direct action of EM1 on striatal DA release was therefore also investigated in an in vitro superfusion system. Although EM1 did not influence the basal release of tritiated DA, it significantly enhanced the transmitter release evoked by electric impulses and pretreatment with L-NNArg resulted in a considerable inhibition of the release elicited by EM1. In conclusion, our endocrine studies suggest an important role of NO in the mediation of the EM1-evoked corticosterone secretion. They also indicate that EM1 activates the HPA axis at a hypothalamic level and dopamine is not involved in this process. In contrast, the behavioural experiments reflect that the locomotor activation induced by EM1 is mediated by NO and dopamine, and the superfusion studies demonstrate that NO transmits the dopamine release enhancing effect of EM1. PMID- 12849759 TI - Voluntary freewheel running selectively modulates catecholamine content in peripheral tissue and c-Fos expression in the central sympathetic circuit following exposure to uncontrollable stress in rats. AB - Modulation of sympathetic drive to the spleen is one potential mechanism whereby physical activity prevents stress-induced splenic immune suppression in rats. The current study tested the hypothesis that voluntary freewheel running reduces peripheral sympathetic drive by modulating stress-induced activity of brain regions synaptically linked to sympathetically innervated peripheral organs, including the adrenals and spleen. To this end, adrenal and splenic catecholamine content and activity of the central sympathetic circuit indexed by c-Fos protein induction, elicited by acute exposure to inescapable tail shock, were measured. Stressor exposure depleted adrenal and splenic norepinephrine content and elicited a robust increase in c-Fos in the brains of sedentary rats. Physical activity status had no effect on adrenal norepinephrine content. Indicative of attenuated sympathetic drive to the spleen, however, 6 weeks of voluntary freewheel running diminished stress-induced splenic norepinephrine depletion, and significantly attenuated stress-induced c-Fos in specific brain regions responsible for sympathetic regulation, including tyrosine hydroxylase immunoreactive neurons of the locus coeruleus, A5 cell group and rostral ventrolateral medulla. Results suggest that voluntary activity attenuates sympathetic drive to the spleen during stressor exposure by selectively modulating stress-induced activity of the central sympathetic circuit. The attenuation of sympathetic responses observed in this study may be one important mechanism for the protective effect of physical activity against stress-related illness and immunosuppression. PMID- 12849760 TI - Long-term potentiation in visual cortical projections to the medial prefrontal cortex of the rat. AB - In order to investigate neural mechanisms by which the prefrontal cortex adaptively modifies its activities based on past experience, we examined whether or not sensory cortical projections to the medial prefrontal cortex support long term potentiation (LTP) in rats. Monosynaptic projections from the secondary visual cortex, mediomedial area (V2MM) to the infralimbic cortex were confirmed by orthodromic as well as antidromic activation of single units. High-frequency stimulation (50 Hz, 2 s) induced LTP (approximately 45% increase over the baseline) in the V2MM projection to the infralimbic cortex. LTP induction in this pathway was completely blocked by an injection (i.p.) of CPP, an N-methyl-D aspartate receptor antagonist. LTP was also induced in the ventral hippocampal projection to the infralimbic cortex by the same high-frequency stimulation. The present results suggest that modification of synaptic weights of afferent sensory cortical projections is one mechanism underlying learning-induced changes in prefrontal cortical neural activities. PMID- 12849761 TI - L-Dopa medication remediates cognitive inflexibility, but increases impulsivity in patients with Parkinson's disease. AB - In the current study we investigated the role of dopamine in attentional and socio-emotional functioning by examining effects of withdrawing dopaminergic medication in patients with Parkinson's disease (PD). Patients "on" medication exhibited abnormal betting strategies on a task of decision-making, reflecting impulsive behaviour and/or delay aversion, whilst the same patients "off" medication exhibited abnormally increased switch costs when switching between two tasks, reflecting attentional inflexibility. Hence, these data replicate and extend previous findings that dopaminergic medication improves or impairs cognitive performance depending on the nature of the task and the basal level of dopamine function in underlying cortico-striatal circuitry. PMID- 12849762 TI - Dissociable effects of dopaminergic therapy on spatial versus non-spatial working memory in Parkinson's disease. AB - There is now evidence for definite and early cognitive deficits in Parkinson's disease (PD), involving, in particular, executive functions and working memory. However, the distinction between visuo-spatial and non-spatial working memory deficits and the impact of dopamine on these deficits are still open to debate. The aim of this study was therefore to investigate cognitive and motor performance in PD patients in two conditional associative learning tasks requiring either spatial or non-spatial visual working memory. The subject had to point to visual targets according to the visual characteristics of memorised visual cues (colour, position and form). To assess the effect of L-dopa therapy, PD patients were studied over two consecutive days: one ON/OFF group of nine PD patients with treatment (ON condition) on the first day and without treatment (OFF condition) on the second day; and another OFF/ON group of nine PD patients tested on reverse. The PD groups were compared to a control group of nine age matched healthy subjects. Our main data demonstrate that: (1) in PD patients with OFF treatment, the response time of manual pointing is increased mainly in the non-spatial working memory task; and (2) in PD patients with ON treatment, either the response time is normal (on the first day) or is increased in both visuo spatial and non-spatial tasks. We suggest that this dissociation between spatial versus non-spatial working memory deficits in non-medicated PD might be related to compensatory mechanisms that occur following fronto-striatal dysfunction. PMID- 12849763 TI - Does the cerebellum contribute to specific aspects of attention? AB - We present data on attentional and neuropsychological functions of 16 patients with focal cerebellar lesions (13 tumours, 3 haematomas) compared to normative test data, and to 11 control subjects matched for age, gender, and years of education. Patients showed distinct deficits in qualitative aspects of a divided attention task, and in a working memory task. Performance in selective attention was unimpaired. The results support the concept that the cerebellum plays a role not only in motor, but also in higher cognitive functions. They are discussed on the basis of the idea that prediction and preparation are fundamental functions of the cerebellum. Therefore, the results confirm the idea that cerebellar lesions lead to reduced performance in specific attention tasks. PMID- 12849764 TI - Cortical and subcortical contributions to the representation of temporal information. AB - Converging evidence suggests that temporal representations of brief durations are derived subcortically. We tested split-brain patient JW in order to investigate whether these representations project bilaterally or unilaterally to cortex. Using visual stimuli to signal time intervals, JW was asked to compare the duration of a pair of standard stimuli that were presented bilaterally with a comparison stimulus that was presented to either the left or right visual field. Assuming the hand of response is controlled by the contralateral cerebral hemisphere, a hand by visual field interaction was predicted if the representation of stimulus duration was restricted to the cerebral hemisphere receiving the lateralized stimulus. However, we failed to observe this interaction for two different ranges of stimulus durations, both in the hundred (Experiment 2) to hundreds (Experiment 1) of milliseconds range. Instead, there was a consistent right hemisphere advantage in task performance. When the task then required a discrimination based on the physical size of the stimuli rather than their duration, an interaction between response hand and visual field was obtained (Experiment 3). Taken together, these results suggest that (1) even though the comparison stimulus was presented unilaterally, the representation of its duration was available to both cerebral hemispheres, and (2) a right hemisphere advantage in psychophysical tasks requiring the comparison of successive stimuli is observed for temporal and non-temporal judgments. PMID- 12849765 TI - The contributions of lesion laterality and lesion volume to decision-making impairment following frontal lobe damage. AB - Lesions to prefrontal cortex (PFC) in humans can severely disrupt everyday decision-making, with concomitant effects on social and occupational functioning. Forty-six patients with unilateral lesions to prefrontal cortex and 21 healthy control subjects were administered three neuropsychological measures of decision making: the Iowa Gambling Task, the Cambridge Gamble Task, and the Risk Task. Magnetic resonance imaging (MRI) scans were acquired from 40 patients, with region of interest (ROI) mapping of prefrontal subregions. The frontal patients showed only limited damage in medial and orbital prefrontal cortex, but greater damage in lateral prefrontal regions of interest. Patients with right frontal lesions preferred the risky decks on the Iowa Gambling Task, and differed significantly from left frontal and control subjects. Within the right frontal group, the preference for the risky decks was correlated with the total lesion volume and the volume of damage outside of the ventromedial prefrontal region. Right and left frontal groups did not differ significantly on the Cambridge Gamble Task or the Risk Task, and performance was not associated with lesion volume. The results indicate a laterality effect on the Iowa Gambling Task, and the contribution of prefrontal regions outside the ventromedial region to task performance. The Cambridge Gamble Task and Risk Task were less sensitive to the effects of unilateral frontal lobe lesions, and may be more selectively associated with ventral prefrontal damage. PMID- 12849766 TI - Differential deficits in expression recognition in gene-carriers and patients with Huntington's disease. AB - Previous studies in symptomatic patients and asymptomatic gene-carriers of Huntington's disease (HD) reported a differential deficit in the recognition of facial expressions of disgust. This impairment may point to involvement of the basal ganglia in the recognition of disgust. In this study, we compared the performance of 20 patients with symptoms of HD, 20 gene-carriers of HD and 20 healthy controls on two tests of facial expressions in order to further investigate the role of the basal ganglia in disgust recognition. Recognition of fear, rather than disgust, was most severely impaired in the patients, who were also impaired at recognising expressions of anger, disgust and sadness. Direct testing for a differential deficit in disgust at the group level (and at the level of individual HD cases) revealed that the patients were in fact significantly more impaired on the other negative expressions than on disgust. The gene-carriers were not impaired on any expression, although there was a trend for the gene-carriers to be poorer at recognising fearful faces than the controls. We argue that the expression recognition performance of the patients and gene-carriers simply reflects differences in task difficulty, rather than dysfunction of any mechanisms dedicated to specific emotions. In contrast to previous studies in patients or gene-carriers of HD, our findings provide no evidence for a role of the basal ganglia in the recognition of disgust and cast doubt on whether results from HD patients and gene-carriers can be used in support of a double dissociation between recognition of disgust and fear. PMID- 12849767 TI - On the automaticity/cerebellar deficit hypothesis of dyslexia: balancing and continuous rapid naming in dyslexic and ADHD children. AB - The present study examined the relationship of reading disability (RD) and attention deficit hyperactivity disorder (ADHD) to balancing problems. In the cerebellar deficit hypothesis of dyslexia of Nicolson et al. [Trends Neurosci. 24 (2001) 508], balancing problems are taken as sign of a cerebellar deficit and were found to be associated with dyslexia. Four groups of 10 children each, representing all combinations of RD (absent versus present) and ADHD (absent versus present), were included. However, poor balancing (assessed both singly and together with a secondary task) was not found to be associated with RD, but with ADHD. In contrast, poor performance on continuous rapid naming tasks (digit and color naming) was found to be associated with RD and not with ADHD. PMID- 12849768 TI - Coordination of fingertip forces during precision grasping in multiple system atrophy. AB - While the pathology and autonomic nervous system components of multiple system atrophy (MSA) have been well described, little is known about the associated motor dysfunction. One prominent feature of MSA is parkinsonism, although ataxias and pyramidal tract signs are frequently present. To investigate the nature of motor deficits in MSA, a natural grip-lift movement requiring a precision grasp was used to examine force coordination. Subjects were asked to grasp an instrumented object using the fingertips of the thumb and index finger and lift it 10 cm above the table surface. Subjects with MSA demonstrated a prolonged duration between object contact and initiation of the lifting drive that increased with the weight of the object. During this period these subjects produced large grasping forces generating a significant portion of the eventual grip force employed to hold the object. In contrast, control subjects generated grip and load forces in parallel after establishing contact with the object. Therefore, subjects with MSA showed a disrupted performance on both the sequential (grasp, then lift) and simultaneous (grip and load force development) portions of this task. Only after initiation of the vertical lifting drive did subjects with MSA generate forces in a similar manner to control subjects. These findings demonstrate that subjects with MSA exhibit a disrupted coordination of grasp and could suggest a general deficit in motor control resulting from multi focal neural degeneration. PMID- 12849769 TI - Intentional and automatic measures of specific-category effect in the semantic impairment of patients with Alzheimer's disease. AB - The breakdown of semantic knowledge relative to living and non-living categories was studied in patients with Alzheimer's disease (AD). The same living and non living items were used in a semantic battery and in a semantic priming paradigm exploring automatic access to the semantic system. Although AD patients showed a semantic deficit on the intentional semantic battery, they demonstrated normal semantic facilitation on the priming task. In the AD group as a whole, the semantic impairment did not preferentially affect the living category either in the intentional or automatic condition. Instead, a prevalent deficit for the living category was found in three AD patients (14% of the group) on the intentional semantic tasks, but not on the automatic one. These findings support the view that the category effect may not be a generalised phenomenon in AD but may be restricted to a limited number of patients. The intentional/automatic dissociation of the semantic breakdown demonstrated by AD patients is discussed in relation to different theories regarding the organisation of semantic memory. PMID- 12849770 TI - Line bisection following hemispherectomy. AB - Two left- and right-hemispherectomized patients with contralateral hemianopia and 20 normal controls were administered a line bisection task. All hemispherectomized patients showed a strong bisection bias towards their blind visual field. This contralateral bias persisted when patients were forced to start scanning within their blind hemifield, supporting the idea of a strategic adaptation of attention towards the blind visual field. In all patients the hemispherectomy was performed as a result of cortical abnormality (congenital or acquired) and therefore early changes in functional cerebral organization may have occurred in these patients. The absence of a neglect-like ipsilateral bias and the presence of a hemianopic-like contralateral bias in line may represent a functional deficit or suggest that plastic changes following hemispherectomy induced an adaptive functional re-organization of spatial attention in both left- and right-hemispherectomized patients. PMID- 12849771 TI - Inhibitory processing following damage to the parietal lobe. AB - We investigated inhibitory properties of spatial attention in a group of four patients with lesions involving the posterior parietal lobe. In a first experiment, a double cue inhibition of return (IOR) procedure was employed. The parietal patients showed an IOR effect only when they had to detect targets that appeared on the contralesional side. In a second experiment, we combined an IOR procedure with a Stroop task [Psychon. Bull. Rev. 8 (2001) 315] to explore the neural basis of "inhibitory tagging" as described by Fuentes et al. [Q. J. Exp. Psychol. Hum. Exp. Psychol. 52 (1999) 149]. The results from the control participants replicated the findings of Vivas and Fuentes, Stroop interference was reduced at the cued location, relative to the uncued location. The parietal patients showed a similar result, but only for contralesional targets. These findings suggest that IOR is modulated by the parietal lobe, and that, through this process, the parietal cortex influences the application of inhibitory tagging to stimuli. PMID- 12849772 TI - Differences in word associations to pictures and words. AB - Normal subjects were asked to produce the "first word that comes to mind" in response to pictures or words that differed with respect to manipulability and animacy. In separate analyses across subjects and items, normal subjects produced a significantly higher proportion of action words (that is, verbs) to pictures as compared to words, to manipulable as compared to non-manipulable stimuli and to inanimate as compared to animate stimuli. The largest proportion of action words was elicited by pictures of non-living, manipulable objects. Furthermore, associates to words matched standard word associates significantly more often than those elicited by pictures. These data suggest that pictures and words initially contact different forms of conceptual information and are consistent with an account of semantic organization that assumes that information is distributed across different domains reflecting the mode of acquisition of that knowledge. PMID- 12849773 TI - Executive skills in Klinefelter's syndrome. AB - Executive skills are those involved in concept formation, problem solving, switching tasks, inhibiting inappropriate responses, initiating rapid and fluent responses, planning and sustained attention. Different patterns of disorder amongst these skills have been found in several developmental abnormalities including autism, attention deficit hyperactivity disorder (ADHD) and Turner's syndrome (TS). This study explored, for the first time, executive skills in children with Klinefelter's syndrome (KS), a sex chromosome abnormality in which there is one or more additional X-chromosomes. Intelligence in KS is normal but there is academic underachievement. A battery of executive tasks was administered, in a series of case studies, to three 10-year-old boys with KS and to controls matched for age, sex and intelligence. The results demonstrate that children with KS have impairments in executive skills. However, the pattern of impairment is task-specific. There is evidence from multiple tasks of impairment in inhibitory skills, for each case of KS. In contrast, concept formation, problem solving, task switching and speeded responding are normal. These results support theories that argue for distinct sub-components of executive skills within development that may develop relatively independently. The results have relevance for modelling both child and adult executive systems. They also confirm that an additional X-chromosome has highly selective effects upon the consequent cognitive phenotype seen in development. PMID- 12849776 TI - Genomic islands and the evolution of catabolic pathways in bacteria. AB - Genes for the degradation of organic pollutants have usually been allocated to plasmid DNAs in bacteria or considered non-mobile when detected in the chromosome. New discoveries have shown that catabolic genes can also be part of so-called integrative and conjugative elements (ICElands), a group of mobile DNA elements also known as genomic islands and conjugative transposons. One such ICEland is the clc element for chlorobenzoate and chlorocatechol degradation in Pseudomonas sp. strain B13. Genome comparisons and genetic data on integrase functioning reveal that the clc element and several other unclassified ICElands belong to a group of elements with conserved features. The clc element is unique among them in carrying the genetic information for several degradation pathways, whereas the others give evidence for pathogenicity functions. Many more such elements may exist, bridging the gap between pathogenicity and degradation functions. PMID- 12849777 TI - Gene transfer occurs with enhanced efficiency in biofilms and induces enhanced stabilisation of the biofilm structure. AB - There has been much interest in bioremediation based on the introduction of bacteria able to catabolise recalcitrant compounds deposited in the environment. In particular, the delivery of catabolic information in the form of conjugative plasmids to bacterial populations in situ has great potential. As most bacteria in the environment live in surface-associated communities (biofilms), the gene transfer systems within these communities need to be better characterised for bio enhancement strategies to be developed. Recent findings suggest that gene transfer does take place within biofilms, but studies also identified limitations and bottlenecks of the process. The dense population structure in biofilms increases plasmid dispersal by conjugation, and the conjugation mechanism itself may stimulate biofilm development. Moreover, DNA release and transformation seem to be part of a biofilm-related life cycle and released DNA stabilises the biofilm structure. Both of these gene-transfer mechanisms may be autocatalytically promoted in biofilms, presenting new possibilities for efficient bio-enhancement strategies. PMID- 12849778 TI - The role of mobile genetic elements in bacterial adaptation to xenobiotic organic compounds. AB - Retrospective studies clearly indicate that mobile genetic elements (MGEs) play a major role in the in situ spread and even de novo construction of catabolic pathways in bacteria, allowing bacterial communities to rapidly adapt to new xenobiotics. The construction of novel pathways seems to occur by an assembly process that involves horizontal gene transfer: different appropriate genes or gene modules that encode different parts of the novel pathway are recruited from phylogenetically related or distant hosts into one single host. Direct evidence for the importance of catabolic MGEs in bacterial adaptation to xenobiotics stems from observed correlations between catabolic gene transfer and accelerated biodegradation in several habitats and from studies that monitor catabolic MGEs in polluted sites. PMID- 12849779 TI - Diversity and dynamics of microbial communities in engineered environments and their implications for process stability. AB - The availability of molecular biological tools for studying microbial communities in bioreactors and other engineered systems has resulted in remarkable insights linking diversity and dynamics to process stability. As engineered systems are often more manageable than large-scale ecosystems, and because parallels between engineered environments and other ecosystems exist, the former can be used to elucidate some unresolved ecological issues. For example, the process stability of methanogenic bioreactors containing well-defined trophic groups appears to depend on the diversity of the functional groups within each trophic level as well as on how these functional groups complement each other. In addition to using engineered systems to study general ecological questions, microbial ecologists and environmental engineers need to investigate conditions, processes, and interactions in engineered environments in order to make the ecological engineering of bioreactor design and operation more practicable. PMID- 12849780 TI - Phytoextraction of metals and metalloids from contaminated soils. AB - The removal of inorganic contaminants by plants is termed phytoextraction. Recent studies have looked at the feasibility of phytoextraction, and demonstrate that both good biomass yields and metal hyperaccumulation are required to make the process efficient. Adding chelating agents to soil to increase the bioavailability of contaminants can sometimes induce hyperaccumulation in normal plants, but may produce undesirable environmental risks. Thus, it is necessary to investigate the mechanisms responsible for hyperaccumulation, using natural hyperaccumulators as model plant species. Recent advances have been made in understanding the mechanisms responsible for hyperaccumulation of Zn, Cd, Ni and As by plants. Attempts to engineer metal tolerance and accumulation have so far been limited to Hg, As and Cd, and although promising results have been obtained they may be some way from practical application. More fundamental understanding of the traits and mechanisms involved in hyperaccumulation are needed so that phytoextraction can be optimised. PMID- 12849781 TI - Elucidating the microbial component of natural attenuation. AB - Microbial reactions are a key determinant in natural attenuation. However, providing unequivocal evidence of the extent of their involvement is challenging. Several approaches are being developed to meet this challenge, including the use of contaminant-specific transformation products, carbon- or hydrogen-based stable isotopic analysis and reactive transport modeling. These approaches emphasize the ongoing need to integrate strategically between temporally and spatially variant geochemical conditions, the ecological characteristics of the resident microbial communities and their resultant pollutant-transformation capabilities. PMID- 12849782 TI - Molecular and physiological approaches to understanding the ecology of pollutant degradation. AB - Pollutant biodegradation in the environment occurs in the context of various interactions among microorganisms. To understand this ecological process, identification of functionally important populations is considered to be the primary step, which can be followed by isolation and laboratory pure-culture studies of the important organisms. Laboratory studies can then proceed to the analysis of in situ activity and interactions with other organisms. Such studies will shape a deeper understanding of the ecology of pollutant degradation and facilitate the development of new bioremediation strategies. PMID- 12849783 TI - Stable-isotope probing of nucleic acids: a window to the function of uncultured microorganisms. AB - Phylogeny based on ribosomal RNA sequences alone is rarely a reliable indicator of microbial function. To circumvent this problem, nucleic acid based techniques have been developed that exploit the physical properties of stable isotopes to study microbially mediated processes within complex environmental samples. Investigations using labelled substrates, or which detect variations in the natural abundance of isotopes, have thus revealed the metabolic function of microorganisms without the need to isolate them in culture. PMID- 12849784 TI - Biotechnological prospects from metagenomics. AB - The recognition that most microorganisms in the environment cannot be cultured by standard methods stimulated the development of metagenomics, which is the genomic analysis of uncultured microorganisms. Two types of analysis have been used to obtain information from metagenomic libraries: a function-driven approach, in which metagenomic libraries are initially screened for an expressed trait, and a sequence-driven approach, in which libraries are initially screened for particular DNA sequences. New antibiotics and enzymes are among the early discoveries from metagenomics. Future refinement of methods that enrich for genes of particular function will accelerate the rate of discovery of useful molecules. PMID- 12849785 TI - Environmental application of array technology: promise, problems and practicalities. AB - Array technology has been applied in environmental research using innovative approaches in gene expression, comparative genomics and mixed community analysis. Greater fundamental understanding of sources of experimental and analytical error in array experiments should facilitate the future application of array technology to environmental analysis. PMID- 12849786 TI - Monitoring bacterial pathogens in the environment: advantages of a multilayered approach. AB - The application of advanced and highly sensitive molecular techniques to the detection of specific bacteria in the freshwater environment is limited, in the first instance, by sampling strategy and sample quality. Further combinations of molecular methods and techniques from apparently unrelated disciplines will ultimately shape the monitoring techniques of the future. PMID- 12849788 TI - Interdisciplinary sciences in the 21st century. AB - With the advance of genome projects the search for the order of cellular processes has become a realistic goal. Yet this order is hidden behind a screen of data, which obscures the actual connectivity of genes and proteins. In order to expose the true nature of these networks it becomes necessary to apply in silico analysis based on certain parallels between biological and complex technical systems. These efforts far exceed current notions of computational biology. PMID- 12849789 TI - Vaccines for the prevention of neglected diseases--dengue fever. AB - Dengue and dengue hemorrhagic fever have spread to all tropical areas of the developing world, but still remain largely neglected diseases. Several promising vaccine candidates in the form of live attenuated and chimeric vaccines have been developed and are currently in human clinical trials. However, significant practical, logistic, and scientific challenges remain before these vaccines can widely and safely be applied to vulnerable populations. Vector control, community education and public health measures must be pursued in parallel with vaccine development. PMID- 12849790 TI - Nanotechnology: convergence with modern biology and medicine. AB - The worldwide emergence of nanoscale science and engineering was marked by the announcement of the National Nanotechnology Initiative (NNI) in January 2000. Recent research on biosystems at the nanoscale has created one of the most dynamic science and technology domains at the confluence of physical sciences, molecular engineering, biology, biotechnology and medicine. This domain includes better understanding of living and thinking systems, revolutionary biotechnology processes, the synthesis of new drugs and their targeted delivery, regenerative medicine, neuromorphic engineering and developing a sustainable environment. Nanobiosystems research is a priority in many countries and its relevance within nanotechnology is expected to increase in the future. PMID- 12849791 TI - Astrobiology, the transcendent science: the promise of astrobiology as an integrative approach for science and engineering education and research. AB - Astrobiology is rapidly gaining the worldwide attention of scientists, engineers and the public. Astrobiology's captivation is due to its inherently interesting focus on life, its origins and distribution in the Universe. Because of its remarkable breadth as a scientific field, astrobiology touches on virtually all disciplines in the physical, biological and social sciences as well as engineering. The multidisciplinary nature and the appeal of its subject matter make astrobiology ideal for integrating the teaching of science at all levels in educational curricula. The rationale for implementing novel educational programs in astrobiology is presented along with specific research and educational policy recommendations. PMID- 12849793 TI - Development of methods of male contraception: impact of the World Health Organization Task Force. AB - OBJECTIVE: To give an historical record of the research of the World Health Organization (WHO) Task Force to develop methods of male contraception; to examine the social, political, medical, pharmaceutical, funding, and other factors that influenced progress; and to suggest reasons why such methods are only now becoming available. DESIGN: Review of basic and clinical research over 30 years. SETTING: Task force of a multinational agency and collaborating agencies. CONCLUSION(S): Through the involvement of many international scientists, the WHO Task Force has uniquely contributed to the exploratory phases of the research in male contraception and by its multicenter contraceptive efficacy studies has accelerated progress towards the ideal hormonal method. Despite an adverse climate involving social and political attitudes, funding constraints, and pharmaceutical industry hesitation, WHO formed coalitions with governments and international agencies to sustain research with results that apply to men in culturally diverse populations and thereby to influence activities across the whole range of global reproductive health and family planning. PMID- 12849794 TI - Does insurance coverage decrease the risk for multiple births associated with assisted reproductive technology? AB - OBJECTIVE: To determine whether insurance coverage for ART is associated with transfer of fewer embryos and decreased risk of multiple births. DESIGN: Retrospective cohort study of a population-based sample of IVF procedures performed in six U.S. states during 1998. SETTING: Three states with mandated insurance coverage (Illinois, Massachusetts, and Rhode Island) and three states without coverage (Indiana, Michigan, and New Jersey). PARTICIPANT(S): Seven thousand, five hundred sixty-one IVF transfer procedures in patients < or = 35 years of age. MAIN OUTCOME MEASURE(S): Number of embryos transferred, multiple birth rate, triplet or higher order birth rate, and triplet or higher order gestation rate. RESULT(S): A smaller proportion of procedures included transfer of three or more embryos in Massachusetts (64%) and Rhode Island (74%) than in the noninsurance states (82%). The multiple-birth rate in Massachusetts (38%) was less than in the noninsurance states (43%). The insurance states all had protective odds ratios for triplet or higher order births, but only the odds ratio (0.2) for Massachusetts was significant. This decreased risk in Massachusetts resulted from several factors, including a smaller proportion of patients with three or more embryos transferred, lower implantation rates when three or more embryos were transferred, and greater rates of fetal loss among triplet or higher order gestations. CONCLUSION(S): Insurance appears to affect embryo transfer practices. Whether this translates into decreased multiple birth risk is less clear. PMID- 12849795 TI - To insure or not to insure: that is the question. PMID- 12849796 TI - To pay or not to pay. PMID- 12849797 TI - Impact of insurance coverage on in vitro fertilization practice patterns: a complex relationship. PMID- 12849798 TI - Insurance is not a magic bullet for the multiple birth problem associated with assisted reproductive technology. PMID- 12849799 TI - Ortho Evra/Evra versus oral contraceptives: follicular development and ovulation in normal cycles and after an intentional dosing error. AB - OBJECTIVE: To compare the effects of the contraceptive patch to oral contraceptives (OCs) on follicular size and incidence of ovulation in normal cycles and after dosing errors. DESIGN: Randomized, open-label. SETTING: Twelve centers. PATIENT(S): One hundred twenty-four ovulatory women. INTERVENTION(S): Subjects received either the patch (groups 1 and 2) or one of three OCs. Correct dosing occurred in cycles 1, 2, 3, and 5. The following dosing errors were planned during cycle 4, a shortened 10-day cycle: [1] patch group 1 subjects wore one patch for 10 consecutive days; [2] for patch group 2 and OC subjects, 7 dosing days were followed by 3 drug-free days. MAIN OUTCOME MEASURE(S): Follicular size, as determined at each cycle by the maximum mean follicular diameter. RESULT(S): After a 3-day dosing error, follicular size was significantly smaller in the patch group (mean, 7.0 mm) vs. each OC group (range of means, 11.8-17.1 mm). Similar results were seen after proper dosing. The incidence of ovulation was significantly lower for the patch users than for women using OCs. CONCLUSION(S): Follicular size and incidence of ovulation were significantly reduced among contraceptive patch users compared with women using OCs in normal cycles and after planned dosing errors. PMID- 12849800 TI - Blood leukocyte subsets are modulated in patients with endometriosis. AB - OBJECTIVE: To determine whether blood leukocyte populations could be modulated in endometriosis. DESIGN: Case-control study. SETTING: Eight clinical institutions of the Montreal area. PATIENT(S): Women with regular menstrual cycles who underwent laparoscopy or laparotomy between 1997 and 2001 and who were not under hormonal treatment for at least 3 months were selected. This study includes 175 cases and 131 controls. MAIN OUTCOME MEASURE(S): The proportion of blood leukocytes expressing markers for T, B lymphocytes, monocytes, or natural killer (NK) cells were compared by flow cytometric analysis between cases and controls. RESULT(S): Age and parity were identified as important confounders. Given that smoking, history of acute infection, and previous use of oral contraceptives strongly correlate with the level of some blood leukocyte populations, these parameters were taken into account in addition with age and parity when the level of blood leukocyte subsets were evaluated in cases and controls. Blood monocytes expressing CD14 and CD44 molecules were increased in patients with endometriosis. Alternatively, B lymphocytes were shown to be significantly decreased in cases compared with controls. CONCLUSION(S): Although these results suggest that endometriosis is associated with some systemic manifestations, the exact role of these modulations remains unclear. PMID- 12849801 TI - Factors that determine discordant outcome from shared oocytes. AB - OBJECTIVE: To investigate the factors that may be related to pregnancy in oocyte recipients who shared oocytes from the same donor and showed discordant pregnancy outcome. DESIGN: Matched pair analysis. SETTING: IVF academic center. PATIENT(S): Five hundred forty-two oocyte donation cycles that shared oocytes from 197 donors with discordant outcome. INTERVENTION(S): Egg donation and embryo transfer. MAIN OUTCOME MEASURE(S): Pregnancy rate and implantation rate. RESULT(S): Three hundred sixty-five matched-paired discordant outcome oocyte recipients were analyzed. Pregnant and nonpregnant recipients were similar in terms of age, serum E(2) levels, endometrial thickness, indications for oocyte donation, and abnormal sperm parameters. No differences were found in the number of oocytes received per patient, fertilization rate, IVF/intracytoplasmic sperm injection (ICSI) distribution, embryo quality, or on embryo transfer difficulty. CONCLUSION(S): Discordant pregnancy outcome could not be explained by the different egg recipients and cycle factors studied. In addition to chance, other factors must be investigated that could explain discordant outcome in egg recipients sharing oocytes from single donors. PMID- 12849802 TI - Legal considerations for cryopreservation of sperm and embryos. AB - OBJECTIVE: To summarize the case law for cryopreservation of sperm and embryos and make recommendations for desirable characteristics that should be included in a cryopreservation clinic's disposition agreements. DESIGN: A literature review of case law and legal review articles was performed. SETTING: Academic research center. PATIENT(S): None. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): None. RESULT(S): Two court cases involving cryopreserved semen and five cases involving cryopreserved embryos are reviewed. CONCLUSION(S): The state of the law surrounding cryopreservation is recent and unsettled. Disposition agreements can provide cryopreservation clinics some degree of certainty in this unsettled area of law. Those drafting cryopreservation disposition agreements are assisted by lessons learned from cryopreservation cases and insight gained from law review articles. It is evident that cryopreservation agreements will be most successful and most likely to be enforced if they are unambiguous, consistent with public policy, and include: a duration provision, each individual's contact information, the individual's provisions for use of their gametes in case of death, and responsibilities of cryopreservation clinic and individuals. Individuals must enter into agreements with a true understanding of the contained provisions. PMID- 12849803 TI - Pronuclear morphology evaluation with subsequent evaluation of embryo morphology significantly increases implantation rates. AB - OBJECTIVE: To elucidate the relative predictive value of implantation markers at different stages of preimplantation development. DESIGN: Correlation of pronuclear morphology with embryo morphology and implantation rates in retrospective and prospective analysis of in vitro fertilization/intracytoplasmic sperm injection (IVF-ICSI) treatment cycles. SETTING: Private infertility center. PATIENT(S): A total of 441 couples undergoing infertility treatment. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Size of pronuclei and distance between them, the number and polarization of nucleolus precursor bodies (NPB) at the one-cell stage, embryo cleavage and fragmentation rates on days 2 and 3, and pregnancy and implantation rates. RESULT(S): Polarization of the NPB in both pronuclei had a statistically significant correlation with normal membrane breakage during ICSI (40%, compared with 33% easy, and 31% difficult membrane breakage) and also with faster cleavage and lower fragmentation rates of embryos. Sixty-one percent of implanting embryos had polarization of the NPB in both pronuclei compared with 37% for all embryos. Larger distance between pronuclei and their unequal size had a statistically significant correlation with slower cleavage and inferior embryo quality. Embryo selection based on only pronuclear morphology or on only day-3 embryo morphology yielded implantation rates of 15.1% and 12.1%, respectively. Embryo selection based on sequential evaluation of both pronuclear morphology and embryo morphology on day 3 resulted in a 21.1% implantation rate. CONCLUSION(S): Polarization of NPB in both pronuclei is as reliable marker of implantation as embryo morphology on day 3. However, pronuclear morphology assessment improves embryo selection only when it is combined with embryo morphology evaluation on day 3. PMID- 12849804 TI - Relationship between peak serum estradiol levels and treatment outcome in in vitro fertilization cycles after embryo transfer on day 3 or day 5. AB - OBJECTIVE: To examine the relationships between peak serum estradiol (E(2)) levels and treatment outcome in in vitro fertilization (IVF) cycles after embryo transfer (ET) on day 3 or day 5. DESIGN: Retrospective analysis of 697 IVF-ET cycles between January 1999 and December 2001. SETTING: A university-affiliated assisted reproduction program. PATIENT(S): Infertile patients undergoing IVF-ET cycles. INTERVENTION(S): Peak E(2) concentration in serum was determined on the day of human chorionic gonadotropin (hCG) administration. The IVF-generated embryos were cultured for 2 days until transfer on day 3. If more than four 8 cell embryos were present on day 3, embryo culture was continued until day 5 for blastocyst transfer. MAIN OUTCOME MEASURE(S): Clinical pregnancy rates. RESULT(S): High peak E(2) levels did not adversely affect treatment outcome. After the cycles were divided according to the day of ET, high peak E(2) levels were associated with improved pregnancy rates after ET on day 5 but not on day 3. CONCLUSION(S): Increasing peak E(2) levels in IVF cycles are associated with improved pregnancy rates after ET on day 5. PMID- 12849805 TI - Monitoring of in vitro fertilization-embryo transfer cycles by ultrasound versus by ultrasound and hormonal levels: a prospective, multicenter, randomized study. AB - OBJECTIVE: To determine whether cycle monitoring using both serum E(2) and ultrasound findings yields superior clinical pregnancy rates during IVF-embryo transfer (ET) compared to monitoring with ultrasound alone. DESIGN: Prospective, randomized, multicenter, patient-blinded study. SETTING: Four assisted conception units in the United Kingdom. PATIENT(S): Two hundred ninety-seven women believed to be normal responders undergoing IVF treatment. INTERVENTION(S): Patients were randomly allocated on day 7 of stimulation to one of the two hCG administration criteria: [1] the E(2)-to-follicle > or =11 mm ratio was between 250 and 500 pmol/L/follicle and at least 2 follicles reached a mean diameter of 18 mm or [2] at least 2 follicles reached a mean diameter of 18 mm and the endometrium thickness was > or =8 mm. MAIN OUTCOME MEASURE(S): Duration and cumulative dose of recombinant human FSH, total number of growing follicles, oocytes retrieved, number and quality of embryos, pregnancy rates, and ovarian hyperstimulation syndrome (OHSS) rates. RESULT(S): Two hundred ninety-seven patients were randomized to one of the two criteria groups. Of these, 288 (97%) received urinary (u)-hCG (143 in group A and 145 in group B). One hundred three women in group A (72%) met both criteria for hCG administration. Pregnancy and OHSS rates were similar (34.3% vs. 31.4% and 4.9% vs. 4.1%, respectively). CONCLUSION(S): The addition of E(2)/follicle criteria to ultrasound monitoring of IVF cycles in normal responders seldom changes the timing of hCG, and does not increase pregnancy rates or the risk of OHSS. PMID- 12849806 TI - Comparison of coasting and intravenous albumin in the prevention of ovarian hyperstimulation syndrome. AB - OBJECTIVE: To compare the efficacy of coasting to prevent ovarian hyperstimulation syndrome (OHSS) with i.v. albumin and to determine if different timing of gonadotropin coasting would affect the IVF outcomes. DESIGN: Retrospective study. SETTING: University hospital-based IVF program. PATIENT(S): One hundred sixty-two women undergoing IVF treatment cycles who were considered to be at high risk for developing OHSS. INTERVENTION(S): Gonadotropin administration was withheld in 60 patients: either coasting occurred before follicular maturation (early coasting subset, n = 28) or follicular maturity was attained before coasting (late coasting subset, n = 32). Outcomes were compared in 102 patients at risk for OHSS who received i.v. albumin on the day of oocyte retrieval. MAIN OUTCOME MEASURE(S): Incidence of OHSS and outcome parameters. RESULT(S): Although the fertilization rates and the incidence of OHSS did not differ significantly between the coasting and i.v. albumin groups, the mean number of oocytes retrieved, the pregnancy rates, and implantation rates were significantly lower in the coasting group. There is no statistical difference in the entire outcome examined and the incidence of OHSS between the early and late coasting subsets. CONCLUSION(S): Coasting is as effective as i.v. albumin in preventing OHSS in high-risk patients but yields inferior pregnancy rates. Early coasting is as successful as late coasting in preventing OHSS with similar IVF outcome. PMID- 12849807 TI - Outcome of intracytoplasmic sperm injection in relation to the meiotic pattern in patients with severe oligoasthenozoospermia. AB - OBJECTIVE: The aim of the study was to evaluate the intracytoplasmic sperm injection outcome in a selected group of patients with oligoasthenozoospermia in relation to the results obtained from their meiotic analysis. DESIGN: Retrospective clinical study. SETTING: An assisted reproduction service and a university department. PATIENT(S): One hundred thirty-seven men with oligoasthenozoospermia grouped in relation to their meiotic pattern. INTERVENTION(S): Two hundred twenty-four intracytoplasmic sperm injection cycles from 137 men with oligoasthenozoospermia in whom diagnostic meiotic analyses had been performed. MAIN OUTCOME MEASURE(S): Fertilization, pregnancy, implantation, and abortion rates. RESULT(S): There were no significant statistical differences in fertilization, pregnancy, implantation, or abortion rates among the three groups studied. CONCLUSION(S): No statistically significant differences in fertilization, pregnancy, implantation, or abortion rates were found in patients with oligoasthenozoospermia in relation to the meiotic pattern. PMID- 12849808 TI - Differential effects of unopposed versus opposed hormone therapy, tibolone, and raloxifene on substance p levels. AB - OBJECTIVE: To evaluate the effects of unopposed therapy (conjugated equine estrogens [CEE]) vs. opposed therapy (CEE and medroxyprogesterone acetate), tibolone, and raloxifene on serum substance p levels. DESIGN: Clinical study. SETTING: University hospital. PATIENT(S): One hundred eight postmenopausal women were assigned to four treatment groups: unopposed hormone therapy (HT) (n = 30), opposed HT (n = 48), tibolone (n = 18), and raloxifene (n = 12). INTERVENTION(S): Conjugated equine estrogens, CEE and medroxyprogesterone acetate, tibolone, and raloxifene were administered orally; blood samples were collected before therapy and 3 months after. MAIN OUTCOME MEASURE(S): Serum substance p levels were measured before and at the end of the third month of the treatment.The serum substance p levels were increased in the unopposed HT group after treatment. On the contrary, substance p levels were decreased in the opposed HT group, in the tibolone group, and in the raloxifene group. CONCLUSION(S): Addition of progesterone (P) to estrogen (E) treatment significantly decreases serum substance p levels. Tibolone and raloxifene exert the same effect. PMID- 12849809 TI - Effect of obesity on recombinant follicle-stimulating hormone absorption: subcutaneous versus intramuscular administration. AB - OBJECTIVE: To determine whether recombinant follicle-stimulating hormone (rFSH) should be administered intramuscularly (i.m.) or subcutaneously (s.c.) to obese women. DESIGN: Randomized, single-center, two-way crossover study. SETTING: Academic clinical research center. SUBJECT(S): Nineteen healthy women of reproductive age with body mass indices of 19.9 kg/m(2)-42.8 kg/m(2). INTERVENTION(S): Leuprolide acetate 3.75 mg i.m. to achieve pituitary down regulation as determined by serum E(2) levels. Subjects were then given a single dose of 300 IU rFSH either i.m. or s.c.. Multiple blood sampling was performed over the next two weeks, and after retreatment with leuprolide, a second 300 IU rFSH dose was given via the other administration route. MAIN OUTCOME MEASURE(S): Serum samples were analyzed in duplicate for follicle-stimulating hormone (FSH) using a standard radioimmunoassay in a single run. Maximum concentrations (C(max)), times to C(max) (T(max)), and extent of absorption (area under curve [AUC]) with i.m. vs. s.c. administration were compared using paired analysis. RESULT(S): Maximal concentrations were achieved within 24 hours with both s.c. and i.m. routes. No significant differences were found in C(max), T(max), or AUC with s.c. vs. i.m. administration. A decline of AUC occurred among subjects of higher body mass index (BMI) with rFSH given either s.c. or i.m.. Subcutaneous administration achieved AUCs comparable to i.m. administration in both normal weight and obese subjects. CONCLUSION(S): Our data indicate that the s.c. administration of rFSH is appropriate for women regardless of body mass. PMID- 12849810 TI - Clinical outcome of using ganirelix acetate versus a 4-day follicular phase leuprolide acetate protocol in unselected women undergoing in vitro fertilization. AB - OBJECTIVE: To compare the clinical outcome of controlled ovarian hyperstimulation (COH) in unselected patients undergoing IVF using multidose ganirelix acetate versus 4 days of administration of leuprolide acetate. DESIGN: Retrospective cohort study. SETTING: A fertility and IVF center. PATIENT(S): Two hundred forty seven women who underwent COH-IVF between April 1, 1999, and January 30, 2001. INTERVENTION(S): Pituitary suppression according to a 4-day follicular phase leuprolide acetate protocol (236 women) or a multidose ganirelix acetate regimen (133 women). MAIN OUTCOME MEASURE(S): Amount of gonadotropin used, days of stimulation, cancellation rate, number of oocytes retrieved, implantation rate, and clinical pregnancy rate. RESULT(S): Compared with leuprolide acetate recipients, ganirelix recipients required significantly less gonadotropin and the mean day of hCG administration was 4 days earlier. Among women younger than 35 years of age, the implantation rate (15% vs. 6%) and the clinical pregnancy rate per initiated and transferred cycle (27% vs. 12% and 32% vs. 15%, respectively) were significantly higher in the ganirelix group than the leuprolide acetate group. CONCLUSION(S): Compared with a 4-day leuprolide acetate protocol, COH-IVF using a multidose ganirelix acetate protocol reduces treatment duration and amount of gonadotropin used. In younger women, the latter protocol is associated with significantly better pregnancy and implantation rates. PMID- 12849811 TI - Predictive usefulness of cycle day 10 follicle-stimulating hormone level in a clomiphene citrate challenge test for in vitro fertilization outcome in women younger than 40 years of age. AB - OBJECTIVE: To determine the usefulness of the day 10 FSH level of a clomiphene citrate challenge test in predicting IVF outcome in women younger than 40 years of age. DESIGN: Retrospective chart review. SETTING: Academic fertility center. PATIENT(S): Three hundred fifty-three women younger than 40 years of age undergoing 483 IVF cycles. MAIN OUTCOME MEASURE(S): Delivery rates. RESULT(S): An inverse relationship was observed between the likelihood of successful pregnancy and both cycle day 3 and cycle day 10 FSH levels. In women with a normal day 3 FSH level, elevated day 10 FSH levels predicted similarly poor IVF outcomes as for women with an abnormal day 3 FSH level. There was no specific threshold FSH level beyond which pregnancies failed to occur. Successful pregnancies were achieved with aggressive stimulation even in women with FSH levels greater than 20 mIU/mL. CONCLUSION(S): The clomiphene citrate challenge test (CCCT) correlates with IVF outcomes in women younger than 40 years of age. Stimulated day 10 FSH levels are strongly predictive of decreased IVF success even when day 3 FSH levels are normal. Results of the CCCT are useful for patient counseling before the IVF cycle and for choosing the optimal gonadotropin regimen. PMID- 12849812 TI - A new model for ovarian follicular development during the human menstrual cycle. AB - OBJECTIVE: To evaluate changes in ovarian follicle dynamics during the human menstrual cycle to test the hypothesis that folliculogenesis occurs in a wave like fashion. DESIGN: Prospective longitudinal study. SETTING: Healthy volunteers in an academic research environment. PATIENT(S): Fifty healthy women of reproductive age (range 19-43 years) with a history of regular menstrual cycles not taking medications known to interfere with reproductive function were evaluated. INTERVENTION(S): Transvaginal ultrasonography was performed daily for one interovulatory interval (IOI). MAIN OUTCOME MEASURE(S): Changes in the diameter and number of follicles > or =5 mm were evaluated. RESULT(S): Sixty eight percent of women exhibited two waves of follicle development during the IOI and 32% exhibited three waves. Waves were characterized by an increase and subsequent decrease in the number of follicles > or =5 mm occurring in association with the growth of > or =2 follicles to > or =6 mm. A day effect and day by wave interaction were detected in the mean diameter of the largest three follicles and the number of follicles > or =5 mm. CONCLUSION(S): The follicular wave phenomenon in women provides a new model for ovarian function during the menstrual cycle and will improve our understanding of the ovarian response to fertility and hormonal contraceptive regimens. PMID- 12849813 TI - Decreased total antioxidant status and increased oxidative stress in women with polycystic ovary syndrome may contribute to the risk of cardiovascular disease. AB - OBJECTIVE: To determine oxidative stress by the level of protein carbonyls and total antioxidant status (TAOS), and whether oxidative stress is associated with increased risk of cardiovascular disease in women with polycystic ovary syndrome (PCOS). DESIGN: Controlled clinical study. SETTING: University hospital. PATIENT(S): Thirty women with PCOS and 31 healthy control women. INTERVENTION(S): Biometric measures and blood samples collection. MAIN OUTCOME MEASURE(S): C reactive protein (CRP), lipid fractions, glucose, protein carbonyls, insulin, and other hormone (gonadotropins, androgens) levels and TAOS were measured. The estimate of insulin resistance was calculated by homeostasis model assessment (HOMA-R). RESULT(S): The women with PCOS had significantly higher serum fasting insulin, CRP, protein carbonyl levels, HOMA-R, LH levels, and LH/FSH ratios than healthy women. However, TAOS was significantly lower in women with PCOS. TAOS was negatively correlated with fasting insulin, HOMA-R, CRP, and protein carbonyls. Fasting insulin was positively correlated with protein carbonyls. High density lipoprotein (HDL) was inversely associated with fasting insulin, HOMA-R, and protein carbonyls. CONCLUSION(S): Increased oxidative stress and decreased antioxidant capacity may contribute to the increased risk of cardiovascular disease in women with PCOS, in addition to known risk factors such as insulin resistance, hypertension, central obesity, and dyslipidemia. PMID- 12849814 TI - Frequency of the T228A polymorphism in the SORBS1 gene in children with premature pubarche and in adolescent girls with hyperandrogenism. AB - OBJECTIVE: Because the metabolic actions of insulin are more impaired than the mitogenic pathways in polycystic ovary syndrome (PCOS), genes coding for proteins involved in insulin-mediated glucose transport can be considered as candidate genes. The sorbin and SH3-domain-containing-1 (SORBS1) gene codes for c-Cbl associated protein (CAP) involved in insulin-mediated glucose uptake. An association study showed that a missense variant of the SORBS1 gene is protective against obesity and diabetes. We tested the hypothesis that the frequency of the protective allele would be decreased in children with premature pubarche and adolescent girls with hyperandrogenism. DESIGN: Association study. SETTING: Academic research environment. PATIENT(S): Children referred for the evaluation of premature pubarche (n = 79), adolescent girls with hyperandrogenism (n = 56), and healthy nondiabetic controls (n = 50). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Frequency of the T228A allele in our patients and the relationship of body mass index to presence or absence of the T228A variant in our patient population. RESULT(S): Using allele-specific restriction fragment length polymorphism, allele frequencies were found to be similar among the premature pubarche, hyperandrogenism, and control groups (6.0%, 4.6%, and 8.0%, respectively). No statistically significant relationships were found between the SORBS1 genotypes and body mass index or hormone status. CONCLUSION(S): This SORBS1 polymorphism does not play a major role in premature pubarche, hyperandrogenism, and/or polycystic ovary syndrome in our patient population. PMID- 12849816 TI - Familial associations in women with polycystic ovary syndrome. AB - OBJECTIVE: To confirm whether there was a familial association between polycystic ovary syndrome (PCOS) and thromboembolic disease, ovarian or breast cancer, diabetes, and heart disease. DESIGN: Cross-sectional study. SETTING: A university hospital in the United Kingdom. PATIENT(S): Two hundred and seventeen women with and without PCOS under the care of the same consultant gynecologist at a teaching hospital. INTERVENTION(S): Questionnaire survey. MAIN OUTCOME MEASURE(S): Prevalence of a personal or positive family history of thromboembolism, ovarian cancer, breast cancer, diabetes, and heart attacks. RESULT(S): In an analysis of the replies from 41 women with PCOS and 66 controls, we found a statistically significant positive family history of breast cancer and heart attacks among women with PCOS. There was no statistically significant difference in the mean age, ethnic origin, or prevalence of a family history of other diseases. CONCLUSION(S): Our results show a positive association between polycystic ovary syndrome and a family history of breast cancer and heart disease. These associations may be genetic in origin, or secondary to a complex interplay of genetic, intrauterine, and environmental factors. More studies are required to confirm these findings and determine the factors that explain these associations. PMID- 12849815 TI - Comparison of various simple insulin sensitivity and beta-cell function indices in lean hyperandrogenemic and normoandrogenemic young hirsute women. AB - OBJECTIVE: To assess and compare various simple insulin sensitivity and beta-cell function indices in lean, hirsute, young women. DESIGN: Prospective study. SETTING: Departments of endocrinology and metabolism at a university and a state hospital. PATIENT(S): Seventy-one hirsute young women were classified as hyperandrogenemic or normoandrogenemic. MAIN OUTCOME MEASURE(S): Insulin sensitivity and beta-cell function indices derived from a single sample and an oral glucose tolerance test (OGTT). RESULT(S): Lean hyperandrogenemic hirsute women have insulin resistance and increased beta-cell function. The most sensitive indices of insulin resistance were total and 1-hour and 2-hour post challenge insulin levels during OGTT. When a cut-off value of 3.2 or greater for homeostasis model assessment of insulin resistance (HOMA-IR) was accepted, 46% of hyperandrogenemic women and 30% of normoandrogenemic women were insulin resistant. Fasting insulin level was best correlated with the fasting insulin resistance index, HOMA-IR, and Quicky index. The HOMA-IR was best correlated with fasting insulin level and the hepatic insulin sensitivity index (ISI(HOMA)). CONCLUSION(S): Insulin levels based on OGTT are the most useful index of insulin resistance and beta-cell function index in hirsute women. The HOMA-IR may be a proposed global test for insulin resistance; it correlated well with both OGTT derived insulin resistance and beta-cell function indices and with global insulin resistance indices derived from a single sample (such as ISI (HOMA), Quicky index, FIRI(-1), fasting Belfiore index, and glucose/insulin ratio). PMID- 12849817 TI - Improved endometrial assessment using cyclin E and p27. AB - OBJECTIVE: To evaluate endometrial expression of cyclin E and p27 in fertile and infertile women. DESIGN: Retrospective clinical study. SETTING: University medical center and private practice. PATIENT(S): Thirty-three fertile volunteers, 83 women seeking infertility treatment, and 23 women undergoing mock cycles. INTERVENTION(S): Endometrial biopsy. MAIN OUTCOME MEASURE(S): Cyclin E and p27 immunohistochemistry. RESULT(S): Glandular cyclin E and p27 expression dramatically changed in intensity and subcellular localization throughout the menstrual cycle. In normal control biopsies, glandular cyclin E progressed from the basal to the lateral cytoplasm (midproliferative phase) to the nucleus (days 18 to 19) and was absent in biopsies after day 20. First appearing on days 17 to 19, p27 was found only in the nuclei. Cyclin E was more frequently seen after day 20 in infertility patients. In the hyperstimulated cycles, staining for cycle E in proliferative samples was more intense than in the natural cycles, but p27 staining was unchanged. CONCLUSION(S): Cyclin E and p27 may be clinically useful markers of development in the endometrium. As cell cycle regulators, cyclins reveal underlying biochemical processes driving endometrial progression and may partly represent the means by which estrogen and progesterone regulate this dynamic tissue. PMID- 12849818 TI - Sexual function in women with complete androgen insensitivity syndrome. AB - OBJECTIVE: To investigate sexual function in women with complete androgen insensitivity syndrome (CAIS) and to investigate the prevalence of factors that might contribute to sexual difficulties. DESIGN: Cross sectional survey and clinical examination. SETTING: Tertiary hospital multidisciplinary intersex clinic and an international peer support group for CAIS. PATIENT(S): Sixty-six adult women with CAIS. INTERVENTION(S): Self-completed survey of sexual function, genital normality perceptions, and compliance and satisfaction with vaginal hypoplasia treatments. Hospital case notes review, and genital examination for prevalence of vaginal and clitoral hypoplasia. MAIN OUTCOME MEASURE(S): Golombok Rust Inventory of Sexual Satisfaction (GRISS) scores of study participants were compared against the scores of the test validation population (as control). In physical examination participants, anatomical dimensions were assessed against published normal values for clitoral and vaginal sizes. RESULT(S): We found that 90% of women with CAIS in this study had sexual difficulties when compared with the general female population, most commonly sexual infrequency and vaginal penetration difficulty; 77% perceived their vagina as small, but on genital examination only 35% had vaginal hypoplasia. CONCLUSION(S): Androgen deficiency leads to sexual problems. Vaginal hypoplasia and negative psychological adaptation to living with an intersex condition are likely to have contributed to the high rates of sexual problems found in this study. Treatments for vaginal hypoplasia need to be evaluated with outcome studies of long-term sexual function, quality of life, and satisfaction. Clinical services for the management of intersex conditions need to be multidisciplinary and aim to optimize the patient's physical and psychological health. PMID- 12849819 TI - Intrauterine insemination results are not affected if Hysterosalpingo Contrast Sonography is used as the sole test of tubal patency. AB - OBJECTIVE: To assess whether women can be assigned to intrauterine insemination (IUI) treatment on the basis of fallopian tubal patency, diagnosed by Hysterosalpingo Contrast Sonography (Hy Co Sy). DESIGN: Case controlled, clinical study. SETTING: Tertiary referral center. PATIENT(S): The cases were consecutive, infertile women who underwent Hy Co Sy and IUI. The control group was women who had IUI over the same period but whose tubal patency was assessed by hysterosalpingogram (HSG) or laparoscopy and dye (lap and dye). INTERVENTION(S): Women with bilateral patency at Hy Co Sy and with unexplained, anovulatory, or male factor infertility underwent IUI using the partner's (IUI-H) or donor's (IUI D) semen. Their outcome was compared with that of cohorts of women who had been examined using HSG or lap and dye. MAIN OUTCOME MEASUREMENT(S): Clinical pregnancy rate per cycle and cumulative pregnancy rates at IUI-H or IUI-D. RESULT(S): The clinical pregnancy rates per cycle at IUI-H or IUI-D did not differ among the three groups. The cumulative pregnancy rates after three cycles of IUI-H were 0.17, 0.15, and 0.17 in the Hy Co Sy, HSG, and lap and dye cohorts, respectively, and 0.69, 0.77, and 0.54 in the same groups after six cycles of IUI D. There were no differences in the clinical characteristics or stimulation regimes used. CONCLUSION(S): Women screened as "normal" by Hy Co Sy may be allocated to treatments that rely on accurate assessment of tubal patency without compromising their chance of conception. PMID- 12849820 TI - Long-term follow-up of cryopreserved hemi-ovary autografts in ewes: pregnancies, births, and histologic assessment. AB - OBJECTIVE: To evaluate a 2-year follow-up of cryopreserved hemi-ovary autografts in ewes. DESIGN: Animal study. SERTTING: University fertility center, Hospices Civils de Lyon; Ecole Nationale Veterinaire de Lyon, INSERM U 418 Hocaron;pital Debrousse, Lyon; and Hopital Edouard Herriot, Lyon, France. PATIENT(S): Grivette ewes. INTERVENTION(S): Recently we reported four pregnancies and six live births after transplantation of frozen-thawed hemi-ovary in six different ewes. The four remaining ewes were monitored for 2 years. After the last birth, the autografted ovary was removed in each ewe during a final laparotomy. The entire grafted ovary was sliced to estimate the remaining primordial follicle population 2 years after grafting. MAIN OUTCOME MEASURE(S): Uterine ultrasound scanning was performed to diagnose pregnancy. Histological assessment of the grafted ovary was performed after delivery. RESULT(S): The four remaining ewes began new gestations. For two of them, this was a second gestation obtained more than 2 years after the autograft. These two ewes delivered male lambs, which died immediately after delivery because of distocia. The lambs were both oversized for gestational age; autopsy found no malformation. A twin pregnancy of a healthy male and a healthy female occurred in May 2002, and a singleton male was born in February 2002. All grafted ovaries showed drastic reduction in follicle population. CONCLUSION(S): Frozen-thawed ovary autograft allowed recovery of fertility a very long time after the procedure despite a drastic reduction in the total number of follicles. PMID- 12849821 TI - Coculture of human oviductal cells maintains mitochondrial function and decreases caspase activity of cleavage-stage mouse embryos. AB - OBJECTIVE: To investigate the mitochondrial function and caspase activity in mouse embryos after human oviductal cell coculture. DESIGN: Experimental laboratory study. SETTING: University gynecology unit. ANIMAL(S): MF-1 (female); BALB/c (male) mice. INTERVENTION(S): Mouse embryos were cocultured with human oviductal cells. MAIN OUTCOME MEASURE(S): Mitochondrial transmembrane potential (Delta psi(m)) and caspase activity. RESULT(S): Compared to embryos after coculture in Chatot-Ziomek-Bavister (CZB) medium supplemented with 0.5 mg/mL of BSA (CZB), Delta psi m of embryos cultured in CZB was significantly lower at the two-cell (CZB, 2.04 +/- 0.412; coculture, 4.34 +/- 0.563) and morula (CZB, 6.06 +/- 0.548; coculture, 7.12 +/- 0.568) stages. Cocultured embryos and in vivo developed embryos had comparable Delta psi m. Caspase activity was not detected in unfragmented cleavage-stage embryos and morula developed in vivo. In vitro cultured morula possessed caspase activity. The activity was significantly reduced in the cocultured morula. CONCLUSION(S): Human oviductal cells maintained the mitochondria function in terms of mitochondrial transmembrane potential and decreased the caspase activity to improve the development of mouse embryo. PMID- 12849822 TI - Role of the plasminogen system in basal adhesion formation and carbon dioxide pneumoperitoneum-enhanced adhesion formation after laparoscopic surgery in transgenic mice. AB - OBJECTIVE: To evaluate the role of plasminogen activator inhibitor-1 (PAI-1), urokinase plasminogen activator (uPA), and tissue-type plasminogen activator (tPA) in adhesion formation after laparoscopic surgery. DESIGN: Prospective, randomized study. SETTING: Academic research center. ANIMAL(S): Seventy female wild-type and transgenic knockout mice for PAI-1 (PAI-1(-/-)), uPA (uPA(-/-)) or tPA (tPA(-/-)). INTERVENTION(S): Standardized lesions to induce peritoneal adhesions were performed during laparoscopy. To evaluate basal adhesions and pneumoperitoneum-enhanced adhesions, the pneumoperitoneum was maintained for 10 minutes or 60 minutes, respectively. Peritoneal biopsy samples were obtained during and after 60 minutes of carbon dioxide pneumoperitoneum. MAIN OUTCOME MEASURE(S): Adhesions were blindly scored after 7 days. Concentrations of PAI-1 and tPA were measured by using enzyme-linked immunosorbent assay. RESULT(S): In PAI-1, uPA, and tPA wild-type mice, pneumoperitoneum enhanced adhesions. Compared with wild-type mice, basal adhesions were fewer in PAI-1(-/-) mice and more in uPA(-/-) and tPA(-/-) mice. Pneumoperitoneum did not enhance adhesions in these transgenic mice. PAI-1 concentration increased after 60 minutes of pneumoperitoneum whereas tPA concentration did not change. CONCLUSION(S): Impaired fibrinolysis increases basal adhesions. The absence of pneumoperitoneum enhanced adhesions in PAI-1(-/-), uPA(-/-), and tPA(-/-) mice and the increase in PAI-1 expression indicate that PAI-1 up-regulation by carbon dioxide pneumoperitoneum is a mechanism of pneumoperitoneum-enhanced adhesion formation. PMID- 12849823 TI - A technique for transplantation of ovarian cortical strips to the forearm. AB - OBJECTIVE: To describe a forearm heterotopic ovarian transplantation technique. DESIGN: Case study. SETTING: Academic medical center. PATIENT(S): One patient with stage IIIB squamous cell cervical carcinoma and one patient with recurrent benign ovarian cysts. INTERVENTION(S): Preparation of thin ovarian cortical slices and transplantation under the skin of the forearm. MAIN OUTCOME MEASURE(S): Follicular development and oocyte retrieval; cyclical estradiol (E(2)) and progesterone (P(4)) production; restoration of serum follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels to reproductive age range. RESULT(S): Both patients were menopausal immediately after oophorectomy. The first patient developed a dominant follicle 10 weeks after transplantation, and her gonadotropin levels decreased to nonmenopausal levels. Percutaneous aspiration of ovarian follicles yielded a metaphase I (M-I) oocyte that was matured to metaphase II (M-II). The first patient's graft was functional for at least 21 months. In the second patient, ovarian follicle development was detected 6 months after transplantation, and periodic menstruation occurred thereafter. Spontaneous ovulation was confirmed by a midluteal increase in her P(4) levels. Menstruation and follicle development continued for more than 2 years after the transplant. CONCLUSION(S): Heterotopic transplantation of ovarian tissue to the forearm is a simple and promising technique to restore ovarian function in women who become menopausal due to chemotherapy, surgery, or radiation. PMID- 12849824 TI - Reliability, feasibility, and safety of minihysteroscopy with a vaginoscopic approach: experience with 6,000 cases. AB - OBJECTIVE: To assess the reliability, feasibility, and safety of lens-based minihysteroscopy. DESIGN: Retrospective comparative study. SETTING: Academic research environment. PATIENT(S): Women with different indications for hysteroscopy. INTERVENTION(S): Six thousand seventeen outpatient diagnostic hysteroscopies with a minihysteroscope (2.7-mm outer diameter [OD] telescope with 3.5-mm OD single-flow diagnostic sheath) and 4,204 with traditional hysteroscope (4-mm OD telescope with 5-mm OD single-flow diagnostic sheath). All hysteroscopies were performed using a vaginoscopic approach and saline to distend the uterus. MAIN OUTCOME MEASURE(S): Rate of successful introduction of the hysteroscope, rate of satisfactory examinations, pain intensity experienced using an arbitrary pain scale (0 = no pain; 1 = low pain; 2 = moderate pain; 3 = severe pain), and number of side effects and complications. RESULT(S): In the minihysteroscopy, group rates of successful introduction and satisfactory examinations were significantly higher than in the traditional hysteroscope group (99.52% vs.72.53% and 98.53% vs. 92.33%, respectively), while pain and vagal reactions were significantly lower (0.10 +/- 0.34 vs.1.09 +/- 0.53 and 2.25% vs.17.12%, respectively). CONCLUSION(S): Hysteroscopy with lens-based minihysteroscopes was easier, less painful, more reliable, and safer than with 5 mm hysteroscopes. Minihysteroscopy with a vaginoscopic approach is a very well tolerated, effective, and safe outpatient procedure. PMID- 12849825 TI - A double-blind randomized trial comparing the Cavaterm and the NovaSure endometrial ablation systems for the treatment of dysfunctional uterine bleeding. AB - OBJECTIVE: To compare two second-generation endometrial ablation systems in women with dysfunctional uterine bleeding (DUB) who want conservative surgical treatment. DESIGN: A double-blind, randomized trial. SETTING: A minimal access gynecological surgery unit in northeast England. PATIENT(S): Fifty-seven women diagnosed with DUB were recruited, with 55 undergoing surgery and completing 12 month follow-up. INTERVENTION(S): Thirty-seven women underwent a NovaSure endometrial ablation, and 18 had a Cavaterm endometrial ablation. Clinical and quality of life data were collected 6 and 12 months after treatment. MAIN OUTCOME MEASURE(S): Amenorrhea, menstrual change, quality of life, sexual activity, patient satisfaction, and procedure acceptability. RESULT(S): Amenorrhea, hypomenorrhea, eumenorrhea, and menorrhagia rates for the Cavaterm and Novasure groups at 12 months were 2/18 (11%) vs. 16/37 (43%); 11/18 (61%) vs. 10/37 (27%); 5/18 (27%) vs. 6/37 (16%); and 0/18 vs. 5/37 (13%), respectively. At 12 months, 83% and 92% of women were either satisfied or very satisfied in the Cavaterm or Novasure groups, respectively. There were no major complications in either group. CONCLUSION(S): Both the Cavaterm and the Novasure endometrial ablation systems are effective in reducing menstrual loss in women with DUB and achieve high rates of patient satisfaction. The Novasure system achieved a statistically significantly higher rate of amenorrhea in this study. PMID- 12849826 TI - Visualization of the epigastric vessels and bladder before laparoscopic trocar placement. AB - OBJECTIVE: Transillumination and laparoscopic visualization are two techniques recommended to minimize the risks of injury to abdominal wall structures during insertion of secondary trocars. This study was designed to determine the effectiveness of these techniques to locate the epigastric vessels and superior bladder margin. DESIGN: Prospective observational. SETTING: Academic medical centers. PATIENT(S): One hundred five women undergoing laparoscopy for tubal sterilization, infertility, pelvic masses, or pelvic pain. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The ability to visualize the superficial and inferior epigastric vessels, and bladder margin; body mass index (BMI; in kilograms per meter squared); and skin color. RESULT(S): Transillumination successfully visualized 64% of superficial epigastric vessels and was less effective both as weight increased (BMI <25 kg/m(2): 86%; BMI = 25-30 kg/m(2): 61%; BMI >30 kg/m(2): 25%) and in dark-skinned women (69%) compared to those with lighter skin (42%). Laparoscopic visualization successfully identified 82% of inferior epigastric vessels and 46% of bladder margins, and was less effective as weight increased. CONCLUSION(S): Transillumination can successfully locate superficial epigastric vessels, and laparoscopic visualization can locate inferior epigastric vessels and the superior bladder margin in the majority of women undergoing laparoscopy. Transillumination is less effective in dark-skinned women, and both techniques are less effective with increasing body weight. PMID- 12849827 TI - Dynamics of endometrial thickness over time: a reappraisal to standardize ultrasonographic measurements in an infertility program. AB - OBJECTIVE: To assess changes in endometrial thickness during different phases of spontaneous uterine contractions in the menstrual cycle. DESIGN: Prospective, longitudinal study. SETTING: Infertility clinic. PATIENT(S): Sixty-eight infertile patients seen for basic assessment between January 30 and July 1, 2002. MAIN OUTCOME MEASURE(S): Endometrial thickness on two-dimensional transvaginal ultrasonography. RESULT(S): Changes in endometrial thickness were seen on transvaginal ultrasonongraphy in 52 of 68 (76.47%) patients during spontaneous uterine contractions throughout the follicular phase of menstrual cycle. Changes involved the myometrium and subendometrium. Changes in thickness ranged from 1.3 mm to 2.8 mm, regardless of the day of study. CONCLUSION(S): Multiple measurements must be made to calculate a mean endometrial thickness that takes into account alterations due to contractions. PMID- 12849828 TI - Endometrial polyp secondary to tamoxifen therapy. PMID- 12849829 TI - An unexpected quadruplet heterotopic pregnancy after bilateral salpingectomy and replacement of three embryos. AB - OBJECTIVE: To report a case of combined intrauterine and interstitial twin pregnancies after bilateral salpingectomy and IVF with replacement of three embryos. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 31-year old woman known to have bilateral salpingectomy for ectopic pregnancies who underwent IVF. INTERVENTION(S): Laparotomy. MAIN OUTCOME MEASURE(S): Postoperation intrauterine monozygotic twins survival and birth. RESULT(S): After removing the interstitial monozygotic twin pregnancy, the patient had an uneventful postoperative course and delivered two healthy girls by cesarean section at 38 weeks' gestation. CONCLUSION(S): Heterotopic pregnancy can still occur in women treated by IVF after bilateral salpingectomy. The early sonography follow-up of IVF pregnancy would be of value because of the reported higher incidence of pathological pregnancies and especially monozygotic twinning. PMID- 12849830 TI - Analysis of semen from patients chronically treated with low or moderate doses of aspirin-like drugs. PMID- 12849831 TI - High survival rate of vitrified human oocytes results in clinical pregnancy. PMID- 12849833 TI - Quorum guidelines for meta-analyses. PMID- 12849834 TI - Quorum guidelines for meta-analyses. PMID- 12849835 TI - The practice of science at its best. PMID- 12849837 TI - The practice of science at its best. PMID- 12849838 TI - Adjunctive use of dexamethasome in Clomid resistant patients. PMID- 12849840 TI - The best-fit model for endometriosis. PMID- 12849842 TI - Fibroid embolization--not without problems. PMID- 12849846 TI - Mitochondrial benzodiazepine receptors regulate oxygen homeostasis in the early mouse embryo. AB - The peripheral benzodiazepine receptor (Bzrp) has been implicated in the control of several processes, including mitochondrial biogenesis and embryo development. The present study examined the impact that specific Bzrp ligands have on oxygen homeostasis in the early mouse embryo. Day 9 embryos at the 16-18 somite pair stage were exposed to standard (21% oxygen) and suboptimal (5% oxygen) oxygen tensions in whole embryo culture. Analysis of gene expression used relative PCR to monitor changes in nuclear respiratory factor-1 (Nrf1), mitochondrial 16S ribosomal RNA (16S rRNA), and genes for several glycolytic enzymes. Ocular development was highly sensitive to periods of hypoxia through a mechanism blocked with the potent Bzrp ligand PK11195. Hypoxia led to a decline of Nrf1 and 16S rRNA levels also through a mechanism blocked with PK11195. Similar activity was observed for FGIN-1-27 whereas Ro5-4864 had contradictory effects. Morpholino based gene knockdown of Nrf1 (anti-NRF1) produced a sequence-specific decrease in 16S rRNA insensitive to PK11195. These functional relationships suggest that Bzrp dependent signals regulate the Nrf1 --> Tfam1 --> mtDNA --> 16S rRNA pathway in response to oxygen levels. The activity of PK11195 most likely has a pharmacodynamic basis with regards to specific embryonic precursor target cell populations, transducing a mitochondrial signal to an Nrf1 response analogous to retrograde regulation in yeast for mitochondria-to-nucleus signaling. PMID- 12849847 TI - Effect of oral dosing vehicles on the developmental toxicity of flubendazole in rats. AB - Flubendazole was suspended in deionized water or olive oil and administered by gavage once daily to pregnant rats on Days 8-15 of pregnancy to examine if the embryolethal and teratogenic doses were affected by the vehicles used. Flubendazole in olive oil caused a statistically significant increase in embryolethality at doses of 7.83 mg/kg per day and higher, with complete resorption in all dams at 31.33 mg/kg per day. When flubendazole was suspended in deionized water, a significant increase in embryolethality occurred only at a maternal dose of 125.32 mg/kg per day. The proportion of litters with anomalous fetuses was significantly increased at doses of 31.33 mg/kg per day and above when flubendazole was administered in deionized water, but increased at doses at four times lower when flubendazole was administered as in olive oil. Administered as a single dose in olive oil on any one of Days 6-12 of pregnancy, a flubendazole dose of 31.33 mg/kg caused significant increases in embryolethality and decreased fetal body weights on Days 7-9, with an 82.7% incidence of embryolethality on Day 8, with complete resorption in 5 of the 8 dams. The critical periods for teratogenic effects were between Days 8 and 11 of pregnancy, with Day 9 being the most critical. Fetuses with gross, skeletal, or internal anomalies were seen in dams given a single dose of as low as 7.83 mg/kg. PMID- 12849848 TI - A case-control teratological study of vaginal natamycin treatment during pregnancy. AB - The aim of this study was to investigate the teratogenicity of vaginal natamycin treatment during pregnancy, because the data of human epidemiological studies have not been published. Pair analysis of cases with congenital abnormalities and matched healthy controls was carried out. The population-based data set of the Hungarian Case-Control Surveillance of Congenital Abnormalities between 1980 and 1996 included as total, 38,151 pregnant women who delivered newborn infants without any defects (control group) and 22,843 pregnant women who had fetuses or newborns with congenital abnormalities. 62 (0.27%) and 98 (0.26%) pregnant women were treated with the natamycin in the case and control groups, respectively (crude OR 1.1 with 95% CI: 0.8-1.5). A teratogenic potential of vaginal natamycin treatment during the second and third months of pregnancy, the critical period of most major congenital abnormalities, was not indicated in the case-control pair analysis (adjusted OR 0.9 with 95% CI: 0.4-1.8). A somewhat higher mean birth weight (72 g) was found in control newborn infants born to mothers with natamycin treatment compared with the data of control newborn infants without this treatment (adjusted P=0.01), though mean gestational age was shorter. The conclusion of this study is that the treatment with vaginal natamycin during pregnancy presents no detectable teratogenic risk to the fetus. PMID- 12849849 TI - Suppression of decidual cell response induced by dibutyltin dichloride in pseudopregnant rats: as a cause of early embryonic loss. AB - In our previous study, dibutyltin dichloride (DBTCl) caused preimplantation embryonic loss and postimplantation embryonic loss in rats following administration at 7.6 mg/kg and above on Days 0-3 and at 3.8 mg/kg and above on Days 4-7 of pregnancy, respectively. This study was designed to assess the effects of DBTCl on uterine function as a cause of early embryonic loss using pseudopregnant rats. DBTCl was given orally to pseudopregnant rats at 3.8, 7.6 or 15.2 mg/kg on pseudopregnant day (PPD) 0-3 or on PPD 4-7. The decidual cell response was induced by bilateral uterine scratch on PPD 4. The uterine weight on PPD 9 served as an index of uterine decidualization. Uterine weight and serum progesterone levels on PPD 9 were significantly decreased after administration of DBTCl at 7.6 mg/kg and above on PPD 0-3 and PPD 4-7. DBTCl had no effect on the serum estradiol levels and number of corpora lutea. Administration of progesterone reversed the suppression of uterine decidualization in rats given DBTCl on PPD 0-3. It can be concluded that DBTCl suppresses the uterine decidual cell response and decreases progesterone levels, and these effects are responsible for early embryonic loss due to DBTCl exposure. PMID- 12849850 TI - Micronucleus formation in 3-day mouse embryos associated with maternal exposure to chlorpyrifos during the early preimplantation period. AB - Chlorpyrifos, an organophosphate pesticide, was evaluated for its ability to induce cytogenetic damage in preimplantation embryos after maternal exposure. Pregnant female mice were intraperitoneally (i.p.) administered a single dose of chlorpyrifos (40 or 80 mg/kg) at 10:00 h on Day 0 of pregnancy. On Day 3 of gestation, blastocysts were collected and evaluated for gross morphology, micronucleus (MN) frequency, and cell number. A significant increase in MN frequency indicating cytogenetic damage was observed in the treatment groups in comparison to control. The MN frequency revealed a clear dose-dependent increase. There was also a significant decrease in the embryo cell number in the 80 mg/kg treated group. A simultaneous decrease in the cell number and increase in MN frequency may reflect an embryonic developmental disadvantage resulting from maternal treatment with chlorpyrifos. PMID- 12849851 TI - Decreased anogenital distance and increased incidence of undescended testes in fetuses of rats given monobenzyl phthalate, a major metabolite of butyl benzyl phthalate. AB - The objective of this study was to determine the adverse effects of monobenzyl phthalate (MBeP), a major metabolite of butyl benzyl phthalate (BBP), on the development of the reproductive system, and to assess the role of MBeP in the antiandrogenic effects of BBP. Pregnant rats were given MBeP by gavage at 167, 250, or 375 mg/kg on days 15-17 of pregnancy. Fetuses were examined on day 21 of pregnancy. Maternal body weight gain and food consumption were significantly decreased at 167 mg/kg and higher. Fetal weight was significantly decreased at 375 mg/kg. A significant increase in the incidence of undescended testes and decrease in the anogenital distance (AGD) and ratio of AGD to the cube root of body weight was found in male fetuses at 250 mg/kg and higher. The AGD and ratio of AGD to the cube root of body weight of female fetuses in the MBeP-treated groups were comparable to those in the control group. The present data indicate that MBeP produces adverse effects on the development of the reproductive system in male offspring and suggest that MBeP may be responsible for the antiandrogenic effects of BBP. PMID- 12849852 TI - Prenatal effects of DuP-697-the irreversible, highly selective cyclooxygenase-2 inhibitor. AB - DuP-697 (5-bromo-2-(4-fluorophenyl)-3-[4-(methylsulfonyl)phenyl]-thiophene), like celecoxib and rofecoxib, is a vicinal diaryl heterocycle highly selective cyclooxygenase-2 (COX-2) inhibitor. The aim of the study was to evaluate prenatal tolerability of DuP-697. The drug was administered orally in Tween 80 water suspension once a day to pregnant Wistar rats, on Days 7-18 of gestation. The initial dose, similar to the rat antipyretic dose, was set at 0.05 mg/kg. The middle dose, 3.5 mg/kg, corresponded to the rat anti-inflammatory and analgesic dose. The high dose was set at 35.0 mg/kg. Control animals received Tween 80 water suspension. On Day 21 of gestation, fetuses were delivered by laparotomy and double stained with alcian blue and alizarin red S or examined using the Wilson technique. Intrauterine growth retardation occurred in the groups exposed to the middle and highest dose of DuP-697. Minimal reactive and degenerative hepatic changes were found in both drug-exposed and control groups. Skeletal malformations were seen occasionally in all drug-treated and control groups. A significant increase in skeleton variations, such as delayed and asymmetrical ossification, was observed in fetuses exposed to the highest drug dose when compared with the control. These changes were not increased (P<0.1) in the middle drug-dose group. The experimentally-derived NOAEL for developmental toxicity was 0.05 mg/kg, and the corresponding LOAEL was 3.5 mg/kg. PMID- 12849853 TI - Estrogenic effects of food wrap packaging xenoestrogens and flavonoids in female Wistar rats: a comparative study. AB - The objective of this study was to compare the estrogenicity of xenoestrogens found in food wrap packaging and phytoestrogen flavonoids. Uterotrophic and vaginal cornification assays were performed on immature and ovariectomized rats. Genistein, bisphenol F, and octylphenol were identified as estrogenic only in immature rats. Using vaginal cornification as a more specific estrogenic parameter, all tested compounds except tangeretin were active in immature rats. While apigenin and kaempferol appeared to have low estrogenic activity, they potentialized the uterotrophic effect of 17beta-estradiol in immature rats. These data showed that (i) phytoestrogens like genistein can be as potent or even more estrogenic than compounds found in food wrap packaging, (ii) immature rats appear to be a more sensitive in vivo model than ovariectomized rats in term of estrogenicity, (iii) the vaginal cornification assay could be a sensitive and useful test to detect weak estrogenic compounds to which humans can be exposed via food. PMID- 12849854 TI - Relationship between sleeping posture and fluctuations in nocturnal scrotal temperature. AB - Elevated testicular temperatures are a potential risk factor for impaired spermatogenesis and reduced semen quality. The development of portable devices for the measurement of scrotal temperatures closely correlating with testicular temperature offered the opportunity to identify periods with high scrotal temperatures during daily life. Notably, sleeping periods covering approximately 1/3 of the day are associated with increased scrotal temperatures to around 36 degrees C. Combining infrared video taping with scrotal temperature profiling, our study clarified that sudden drops of scrotal temperature during sleep are the consequence of changes of posture. Nocturnal scrotal temperatures were significantly higher during sleeping on the side compared with periods lying on the back (+0.65 degrees C, right scrotum, P<0.001; +0.54 degrees C, left scrotum, P<0.001). Median values of nocturnal scrotal temperatures did not significantly differ between 11 volunteers with normozoospermia and 22 men with oligozoospermia. Furthermore, continued scrotal temperature profiling during the following day only showed significantly higher values among men with oligozoospermia than among volunteers for the left scrotal side (+0.70 degrees C, P<0.05), but not for the right scrotal side (+0.39 degrees C, P>0.05). This difference could not be attributed to higher scrotal temperature values during periods with physical inactivity (median in both groups above 36 degrees C) but the duration of physical inactivity in men with oligozoospermia was 50% longer than in volunteers. Thus, in the setting of the present study it was not possible to demonstrate a close correlation between scrotal temperature and semen quality. PMID- 12849855 TI - The presence of environmental pollutants in the semen of farm animals (bull, ram, goat, and boar). AB - Organochlorine pesticides and polychlorinated biphenyls are widely used in agriculture and industry, respectively, and may affect male reproduction function. Although several pollutants have been detected in human semen, similar studies in farm animals have not appeared. In the present study, the semen of bulls, rams, goats, and boars was assayed for the organochlorine pesticides hexachlorobenzene (HCB), hexachlorocyclohexane (HCH, alpha-, beta-, and gamma isomers), dieldrin, and heptachlor epoxide, for DDT-related chemicals (o,p'-DDE, p,p'-DDE, p,p'-DDD, p,p'-DDT), and for the PCBs congeners (PCB-52, -101, -138, 150, and -180). In all species of farm animals, the most frequently detected pollutants were p,p'-DDE (80-100% of samples), HCB (73.9-100%), and gamma-HCH (69.6-100%). Species differences in the concentrations of HCB, alpha-, beta-, and gamma-HCH, dieldrin, p,p'-DDE, p,p'-DDD, and PCBs were noted as well as differences in the concentrations of some isomers of HCH, DDT-related chemicals, and PCB congeners in the same species. PMID- 12849856 TI - Lead and cadmium concentration in the seminal plasma of men in the general population: correlation with sperm quality. AB - The concentration of lead and cadmium in the seminal plasma of men in the general population, including fertile and infertile subjects, was measured. Semen samples were categorised as demonstrating oligospermia (sperm concentration < 20 x 10(6)/ml), asthenospermia (<50% motilesperm), oligoasthenospermia (a combination of the two criteria), and azoospermia (no sperm). An increase in lead and cadmium levels was observed in infertile men and there was a significant negative correlation of cadmium and lead semen concentration with sperm motility and sperm concentration in oligoasthenospermic men. PMID- 12849857 TI - Semen quality of Indian welders occupationally exposed to nickel and chromium. AB - The semen quality of 57 workers from a welding plant in South India and 57 controls was monitored. Blood nickel and chromium concentrations were determined by ICP-MS. Analysis of semen samples was performed in accordance with World Health Organization criteria. The blood level of nickel and chromium for the 28 exposed workers was 123.3 +/- 35.2 and 131.0 +/- 52.6 microg/l, resepctively, which was significantly higher than the 16.7 +/- 5.8 and 17.4 +/- 8.9 microg/l for the control group (n=27). Sperm concentrations of exposed workers were 14.5 +/- 24.0 millions/ml and those of the control group were 62.8 +/- 43.7 millions/ml. Rapid linear sperm motility was decreased in exposed workers compared to controls. There was a significant positive correlation between the percentage of tail defects and blood nickel concentration in exposed workers. The sperm concentration showed a negative correlation with blood chromium content in workers. More abnormal characteristics were found in the semen of exposed workers. Semen abnormalities correlated with the number of years of exposure to welding fumes containing nickel and chromium. PMID- 12849858 TI - Bisphenol A-induced apoptosis of cultured rat Sertoli cells. AB - Bisphenol A (BPA) was examined for its effects on cultured Sertoli cells established from 18-day-old rat testes. We demonstrated that exposure of cultured Sertoli cells to BPA decreased the cell viability in a dose- and a time-dependent manner and that exposure to BPA brought about morphologic changes of the cells, such as membrane blebs, cell rounding, cytoskeletal collapse, and chromatin condensation or fragmentation, all of which conform to the morphologic criteria for apoptosis. Immunocytochemistry showed that active caspase-3, a major execution caspase, was expressed in round Sertoli cells positively labeled by the TUNEL method. Co-localization of active caspase-3 and aggregated actin fragments was also observed in the round Sertoli cells. Theses results suggest that BPA induces cell death of Sertoli cells by promoting apoptosis. Apoptosis-inducing cell death was observed in cells exposed to 150-200 microM BPA, while BPA at <100 microM had only slight cytotoxic effects on the cells. PMID- 12849859 TI - Effect of PCB 126 and PCB 153 on incidence of apoptosis in cultured theca and granulosa cells collected from small, medium and large preovulatory follicles. AB - The aim of the presented study was to evaluate the effects of PCB 126 and PCB 153 on granulosa and theca cell apoptosis. Granulosa and theca cells were collected from small, medium, and large preovulatory porcine follicles and cultured as monolayers. Cells were initially cultured for 24 h to allow attachment to the plates. Media were changed and 100 pg/ml PCB 126 or 100 ng/ml PCB 153 were added. After 48 h, granulosa and theca cells were fixed for assessment of the number of apoptotic cells utilizing a Hoechst staining technique or frozen for measurement of caspase-3 activity. Media were collected for testosterone concentration analysis from theca cell cultures or estradiol from granulosa cell cultures. Neither PCB 153 nor PCB 126 had an effect on testosterone secretion by theca cells collected from small and medium size follicles, while both PCBs decreased testosterone secretion by large follicles. The decrease in testosterone secretion by large follicles under the influence of both PCBs was paralleled by a suppression of caspase-3 activity and a decreased incidence of apoptotic bodies. Neither of the PCBs had an effect on estradiol secretion by granulosa cells collected from small and medium size follicles, while both PCBs increased estradiol in granulosa cells collected from large follicles. PCB-associated increased estradiol secretion by granulosa cells collected from large follicles was accompanied by suppression of caspase-3 activity and a decreased incidence of apoptotic bodies. In conclusion, we have presented evidence that in preovulatory follicles PCBs inhibit both theca and granulosa cells apoptosis. Therefore, an exposure to PCBs may cause alterations in the pattern of terminal differentiation of follicles and attenuate spontaneous elimination of atretic follicles. PMID- 12849862 TI - Trophoblastic remodeling in normal and preeclamptic pregnancies: implication of cytokines. AB - OBJECTIVE: To summarize the recent knowledge on the implications of placenta and cytokines in normal and preeclamptic pregnancies. DATA SOURCES: A literature search was conducted of applicable articles related to interactions between trophoblast and cytokines in generating preeclampsia. CONCLUSIONS: The initiating event in preeclampsia has been postulated to be the reduced uteroplacental perfusion as a result of abnormal extravillous cytotrophoblast invasion and remodeling of the uterine spiral arteries. Focal ischemia and hypoxia, deportation of hypoxemic trophoblast cells and abnormal expression of various placental biologic molecules, particularly the cytokines, are thought to lead to widespread dysfunction of the maternal vascular endothelium resulting in overproduction of endothelin and thromboxane, enhanced vascular sensitivity to angiotensin II, and reduced secretion of vasodilators such as nitric oxide and prostacyclin. These alterations, in turn, cause hypertension, proteinuria and edema, and pathologies in many organ systems (kidney, lung, liver, brain). PMID- 12849863 TI - Comparison of point-of-care-testing glucose meters with standard laboratory measurement of the 50g-glucose-challenge test (GCT) during pregnancy. AB - OBJECTIVES: Although glucose meters are well-established instruments for self monitoring blood glucose levels, diagnostic and screening procedures should be performed using standard laboratory methods. In addition to standard laboratory methods, HemoCue is authorized for screening and diagnostic purposes in Germany. The rapid development of other glucose meters makes it necessary to re-evaluate this recommendation. Our objective was to test the usefulness of glucose meters in screening pregnant patients for gestational diabetes. METHODS: The 50-g glucose challenge test was administered to one hundred and ninety-three pregnant patients whose blood glucose levels were then simultaneously measured with five portable meters and the HemoCue. The results were compared to our standard method (Hexokinase). A cut-off of 7.8 mmol/L was used and sensitivity, specificity, accuracy, the Youden index, and the Kappa index were calculated. The tests were performed by well-trained personnel (C.D. and U.S.). RESULTS: 1212 measurements were performed on 193 patients. All glucose meters showed a very good correlation (r > 0.90). None of the measurements showed an extreme deviation necessitating the error grid analysis. The GlucoTouch (5.93% +/- 7.4) and the HemoCue (-9.04% +/- 5.9) showed a mean deviation greater than 5%. None of the meters had a mean deviation greater than 10%. The accuracy fluctuated between 0.85 and 0.94. The Kappa index was between 66 to 85. In our clinical trial, the Accu-Chek, Glucotouch, OneTouch, and Precision demonstrated greater accuracy and a higher Kappa index than the HemoCue. CONCLUSIONS: Our data showed good concordance in statistical and clinical parameters for most of the six glucose meters. The HemoCue, recommended as a standard method in several countries, did not show better concordance than most of the tested glucose meters. When used by well trained personnel, the accuracy of the Accu-Chek, Glucotouch, One-Touch, and Precision was acceptable for use in gestational diabetes screening. PMID- 12849864 TI - Enzymatic diagnostic test for Muscle-Eye-Brain type congenital muscular dystrophy using commercially available reagents. AB - OBJECTIVES: Mutations disrupting the interaction of extra-cellular ligands and alpha-dystroglycan are responsible for an etiologically heterogeneous group of autosomal recessive congenital muscular dystrophies (CMD) that can have associated brain and eye abnormalities. The objective is to develop a diagnostic test for one of these CMDs, Muscle-Eye-Brain disease (MEB), due to mutations in the gene encoding Protein O-Mannosyl beta-1,2-N-acetylglucosaminyltransferase 1 (POMGnT1). DESIGN AND METHODS: POMGnT1 enzyme activity was determined in extracts of muscle biopsies from four MEB patients and various controls using commercially available reagents. RESULTS: All four MEB muscle samples showed a highly significant decrease in POMGnT1 activity relative to controls. CONCLUSIONS: The assay of POMGnT1 activity in MEB muscle provides a rapid and relatively simple diagnostic test for this disease. CMDs associated with brain malformations such as MEB, WWS and FCMD are heterogenous in clinical presentation and on radiologic examination, suggesting that POMGnT1 assays of muscle biopsies should be used as a screening procedure for MEB in all CMD patients associated with brain malformations. PMID- 12849865 TI - Plasma glutathione peroxidase in healthy young adults: influence of gender and physical activity. AB - OBJECTIVES: Oxidative stress is implicated in the pathophysiology of many cardiovascular diseases. Plasma glutathione peroxidase (pl x GPx, GPx3) is an antioxidant enzyme found in the extracellular fluid. This study aimed to determine reference values for serum GPx3 concentration and GPx activity in young healthy males and females of similar aerobic fitness and to determine the impact of acute physical activity on serum GPx. DESIGN AND METHODS: 52 young, healthy but not physically trained subjects (24 male, 28 female; age = 20.4 +/- 0.7 yr, cycling VO(2 max) = 39.2 +/- 1.6 mL/kg/min; mean +/- SE) participated in this study. An independent group of 18 subjects participated in an acute, 90 min bout of 50% VO(2 max) cycling exercise. Serum GPx activity and GPx3 protein levels, as well as estradiol and 8-iso- prostaglandin F(2alpha) (8-iso-PGF(2alpha;) an index of lipid peroxidation) were determined. RESULTS: Females had significantly higher serum GPx3 concentration (29.1 +/- 1.6 vs. 24.2 +/- 1.3 mg/L, p < 0.01) and serum GPx activity (256.4 +/- 10.4 vs. 222.8 +/- 15.6 U/L, p < 0.05) than males; specific activity (U/mg) was not different between genders. There was no significant gender difference in 8-iso-PGF(2alpha). No significant correlation was found between either GPx activity or GPx3 concentration and serum estradiol or VO(2)(max). The acute, prolonged, mild intensity exercise did not affect serum GPx activity or 8-iso-PGF(2a) levels in males or females. CONCLUSIONS: The results of this study suggest that in a young, healthy but not physically well trained population females are endowed with slightly higher serum GPx3 concentrations and GPx activities than males, but the functional significance of this has not been established. Furthermore, the results indicate that serum GPx levels are not associated with aerobic fitness level, or serum estradiol concentration and that acute, prolonged, mild exercise does not affect the activity of serum GPx in this population. PMID- 12849866 TI - DNA as an enzyme. The effect of imidazole derivatives as cofactors and metal ions as activators or inhibitors. AB - OBJECTIVE: A highly sensitive spectrofluorometric method using dichlorofluorescin (LDCF) was employed to study the rate of electron transfer reaction in presence of DNA and some imidazole derivatives. RESULTS: In our experiments, DNA possessed a unique enzyme like catalytic function in oxidative conversion of nonfluorescent LDCF to fluorescent dichlorofluorescein (DCF). The rate enhancement was associated with the turn over constant: k(p) = 10 s(-1) for DNA and cinnamoyl imidazole as a cofactor. A biphasic saturation curve was observed when the reaction velocities were measured at fixed concentrations of DNA and variable amounts of carnosine. Each of the biphasic trends gave the Scatchard values of V(m1)/K(m1) = 3.1 x 10(-5) and V(m2)/K(m2) = 5.1 x 10(-6) with K(m1) = 2.7 x 10( 5) M and K(m2) = 4.2 x 10(-4) M for carnosine. Although Ni (II) and Pb (II) induced inhibition in the rate of electron transfer reaction in presence of DNA and cinnamoyl imidazole or carnosine, metal ions such as Mg (II), Cd (II), Zn (II) and Fe (II) caused activation of DNA. The rates of the reactions showed strong dependency on electronegativity and conductivity of metal ions, namely the increase in activity of DNA in presence of each metal ion correlated inversely with the electronegativities of the metal and was also related directly to the conductivities of individual metal. These effects were observed both in activation and also inhibition of DNA reaction. Imidazole compounds, e.g., Histidine, N-trans cinnamoyl imidazole and imidazole along with Cd (II) produced further rate enhancement. The increase was several times greater with N-trans cinnamoyl imidazole. CONCLUSIONS: This effect could provide additional evidence for the importance of an intermediary cofactor that could facilitate the transfer of the electron from the reaction site to the DNA conductive chord. This was most guaranteed by the conjugated system provided by a compound such as N-trans cinnamoyl imidazole. PMID- 12849867 TI - Helicobacter pylori is associated with modified lipid profile: impact on Lipoprotein(a). AB - OBJECTIVES: Helicobacter pylori is a controversial risk factor for atherosclerosis. We investigated whether the bacterium persistent inflammation or the expression of the cytotoxin-associated gene A (CagA) may affect serum lipids as well as Lipoprotein(a). DESIGN AND METHODS: Two hundred-eleven healthy volunteers were evaluated for lipids and Lipoprotein(a). Helicobacter pylori was characterized by Urea Breath Test and IgG-anti-CagA. apo(a) Kringle-IV polymorphism was genotyped. RESULTS: Prevalence of the infection was 72%; 43% of subjects expressed CagA reactivity. Infected subjects showed increased levels of cholesterol, LDL-cholesterol, and cholesterol/HDL-cholesterol atherogenic index. Association with the Helicobacter pylori CagA(-) strains persisted after the adjustment for covariates. Significant difference between infected and uninfected subjects was found in Lipoprotein(a) levels. This difference did not arise from the Kringle-IV genotype. CONCLUSIONS: The infection per se significantly modified serum lipid and Lipoprotein(a) concentrations. CagA does not seem to be a reliable marker of pathogenicity for the atherogenic complications of H. pylori infection. PMID- 12849868 TI - Insulin, proinsulin and insulin resistance status in relation to lipid profiles among school children in Taiwan-The Taipei Children Heart Study. AB - PURPOSE: Evidence suggests that there may be a metabolic syndrome characterized by hyperinsulinemia or insulin resistance associated with increased cardiovascular disease risk. The purpose of this study is to evaluate insulin, proinsulin or insulin resistance to determine which is the best parameter to predict lipid profiles among children in Taiwan. METHODS: After multi-stage sampling, we randomly included 852 school children (415 boys and 437 girls) with a mean age of 13 yr in this study. We measured insulin and intact proinsulin levels by RIA (<0.2% cross-reactivity) and estimated insulin resistance index (IRI) using the homeostatic model assessment (HOMA) method. We used standard methods to measure atherosclerotic lipid profiles including total cholesterol (CHOL), triglyceride (TG), HDL-C, apolipoprotein A (ApoA), apolipoprotein B (ApoB), and lipoprotein[a] and calculated LDL-C and TCHR (total cholesterol to HDL-C ratio) levels. RESULTS: Girls had higher CHOL, LDL-C, ApoA and ApoB levels than boys (p < 0.001). There was no significant difference in insulin, proinsulin and IRI status between boys and girls. Among boys, insulin, proinsulin and IRI were positively correlated with TG, ApoB and TCHR and negatively related to HDL C. Among girls, these associations were attenuated and became insignificantly for TCHR and HDL-C. After adjusting for potential confounders, IRI and insulin were still positively associated with TG and ApoB levels and negatively associated with HDL-C in boys. However, in girls, proinsulin and insulin were positively associated with TG only. Finally, in the stepwise regression analyses, IRI was a better predictor of TG, HDL-C, and ApoB than insulin or proinsulin in boys. However, in girls, proinsulin was a stronger predictor than insulin or IRI for TG and TCHR. CONCLUSION: From this study, we found that IRI (in boys) and proinsulin (in girls) levels are generally more significant and stronger parameters than insulin for predicting lipid profiles among children in Taiwan. PMID- 12849869 TI - Lectin and serum-PSA interaction as a screening test for prostate cancer. AB - OBJECTIVES: The present investigation was designed to distinguish prostate cancer and benign prostate hyperplasia by lectin-prostate specific antigen (PSA) binding. DESIGN AND METHODS: The quantitative precipitin method of concanavalin A (Con A)-carbohydrate interaction was explored with the serum PSA of patients suffering from prostatic complications. RESULTS: The carbohydrate content in the precipitate after binding of Con A with serum PSA of prostate cancer was significantly lower than that of benign prostate hyperplasia. This may be due to altered sugar chain structure or less glycosylation of PSA in prostate cancer. CONCLUSIONS: We conclude that a serum value <3.0 microg/ml of the carbohydrate content of Con A-PSA precipitate indicates strong suspicion for prostate cancer and this cut off level is effective in reducing the rate of unnecessary biopsies in men with total PSA value between 4.0 to 10.0 ng/ml. PMID- 12849870 TI - Involvement of oxidative and nitrosative stress in promoting retinal vasculitis in patients with Eales' disease. AB - OBJECTIVES: Eales' disease (ED) is an idiopathic retinal vasculitis condition, which affects retina of young adult males. The histopathological hallmark in ED is the adhesion of leukocytes to the endothelium and the infiltration of these cells into the retinal parenchyma. Phagocyte generated free radicals have been implicated in mediating tissue damage associated with various inflammatory vasculopathies. In the present study, we have investigated the possible role of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in causing retinal tissue damage in ED. DESIGN AND METHODS: 35 patients with ED and 20 healthy control subjects were included in the study. Monocytes (MC) were separated from peripheral blood of the respective study participants. Inducible nitric oxide synthase (iNOS) protein expression was assessed using Western blot and 3 nitrotyrosine (3NTYR) by reversed phase high performance liquid chromatography (RP HPLC). Thiobarbituric acid reactive substances (TBARS) were determined by measuring malondialdehyde (MDA) formed. Superoxide dismutase (SOD) activity was assayed based on the ability of SOD to inhibit auto-oxidation of epinephrine. Iron, copper and zinc content were determined using Atomic Absorption Spectrophotometer. Immunolocalization of iNOS and 3NTYR was performed on the surgically excised epiretinal membranes (ERM) from patients with ED. RESULTS: There was a significant increase in the expression of iNOS, as well as 3NTYR accumulation, diminished SOD activity, elevated lipid peroxides, iron, copper and decreased zinc content in the MC of patients with ED when compared with healthy control subjects. The elevated levels of ROS and RNS products correlated with diminished antioxidant status in patients with ED. Strong immunoreactivity for iNOS and 3NTYR was observed in inflammatory cells and endothelial cells in ERM obtained from patients with ED. CONCLUSIONS: Our findings from this study clearly reveal the involvement of RNS and ROS in the development of retinal vasculitis in ED. Based on our present study and earlier studies we confirm the role of free radicals in mediating retinal tissue damage in ED. Hence we believe selective inhibition of iNOS or supplementation with antioxidants vitamin E and C might be beneficial in controlling retinal vasculitis in patients with ED. PMID- 12849871 TI - Persistence of vitamin B12 insufficiency among elderly women after folic acid food fortification. AB - OBJECTIVE: To estimate the associated risk of folate and vitamin B12 (B12) insufficiency, as well as vitamin repletion, following folic acid food fortification. DESIGN: Retrospective cross-sectional study over a 5-year period. SETTING: Two large laboratory databases in the provinces of Ontario and British Columbia, Canada. PARTICIPANTS: Canadian women aged 65 years and over who underwent concomitant clinical testing of serum folate and B12 during the pre fortification period of January 1996 to December 1997 in Ontario (n = 733) and British Columbia (n = 3839), and in the near-complete post-fortification period of January 1998 to December 2000 in Ontario (n = 4415) and British Columbia (n = 6677). MEASUREMENTS: Geometric mean concentrations of serum folate and B12 before and after folate fortification. Prevalence ratios (PR) were used to separately compare the post- and pre-fortification period rates of folate deficiency (below 6.0 nmol/L); B12 insufficiency (below 150 pmol/L); and B12 insufficiency in combination with supraphysiological concentrations of serum folate (above 45 nmol/L). RESULTS: The mean baseline folate and B12 concentrations were similar between provinces. Using the combined provincial data, the mean serum folate concentration increased by 64% after fortification, from 14.8 to 24.2 nmol/L (p < 0.001). The average B12 concentration increased from 280 to 300 pmol/L, which was more pronounced in BC (p < 0.001) than in Ontario (p = 0.16). The prevalence of folate deficiency declined from 6.3% to 0.88% after fortification (PR 0.14, 95% confidence interval [CI] 0.11-0.18), while the decline in B12 deficiency was less pronounced (PR 0.78, 95% CI 0.71-0.86). CONCLUSIONS: The prevalence of combined B12 insufficiency with supraphysiological concentrations of serum folate increased from 0.09% pre-fortification to 0.61% post (PR 7.0, 95% CI 2.6-19.2). The introduction of folic acid food fortification was associated with a substantial improvement in the folate status of Canadian women aged 65 years and older, paralleled by a large decline in the rate of folate deficiency. Improvement in the B12 status of these women was far less pronounced. Because the prevalence of combined B12 insufficiency and supraphysiological concentrations of serum folate may have increased with folic acid food fortification, consideration should be given to confirming this finding, and possibly, to the addition of B12 to folate fortified foods. PMID- 12849872 TI - Evaluation of glucose-6-phosphate dehydrogenase activity in two different ethnic groups using a kit employing the haemoglobin normalization procedure. AB - AIM: Correct evaluation of Glucose-6-Phosphate Dehydrogenase (G-6-PD) activity of two ethnic groups using a fully quantitative kit with a simultaneous Hemoglobin Normalization (Hb Normalization) procedure. DESIGN AND METHODS: Two groups of mothers and their healthy full term newborns of Greek (n = 1.166) and Albanian (n = 818) origin were tested for their G-6-PD activity employing a direct normalization protocol. RESULTS: Greek mothers and newborns showed a higher prevalence for G-6-PD deficiency as compared to those of Albanian origin. Males of G-6-PD deficient mothers confirmed the efficacy of the method. CONCLUSION: A fully quantitative G-6-PD kit employing Hb Normalization is essential for the correct classification of G-6-PD activity, both in male and female subjects. PMID- 12849873 TI - Age-related increases in plasma malondialdehyde and protein carbonyl levels and lymphocyte DNA damage in elderly subjects. AB - OBJECTIVES: Increased oxidative stress has been hypothesized to play an important role in the aging process. A role for oxidative damage in normal aging is supported by studies in experimental animals, but there is limited evidence in humans. To investigate the relationship between the oxidative stress and aging in humans, we determined lipid and protein oxidation in plasma as well as DNA damage in lymphocytes in young and elderly subjects. DESIGN AND METHODS: 55 healthy subjects were divided into young (21-40 years) and elderly (61-85 years) groups. Plasma malondialdehyde (MDA), protein carbonyl (PC) levels, and grade of DNA damage in lymphocytes using comet assay as well as total ferric reducing antioxidant power (FRAP) in plasma were determined in young and elderly subjects. RESULTS: Plasma MDA and PC levels were found to be increased in plasma of elderly subjects as compared to young subjects. Increases in endogenous and H2O2-induced DNA damage were also observed in lymphocytes of elderly subjects. In addition, we detected a significant decrease in FRAP values in elderly subjects. Plasma MDA, PC levels and endogenous and H2O2-induced DNA damage were positively correlated with aging, but negatively with FRAP values. CONCLUSION: We evaluated MDA, PC levels and lymphocyte DNA damage altogether in both young and elderly subjects for the first time. The results of this study strongly support the presence of increased oxidative stress in elderly subjects. PMID- 12849874 TI - An amplification-free hybridization-based DNA assay to detect Streptococcus pneumoniae utilizing the up-converting phosphor technology. AB - OBJECTIVES: Development of a generally applicable sensitive hybridization-based assay devoid of any target amplification for the detection and identification of (pathogenic) bacterial and viral species. DESIGN AND METHODS: Using a sandwich hybridization format, the presence of a species-specific nucleic acid sequence is detected by means of Lateral Flow (LF) and Up-converting Phosphor Technology (UPT, a luminescent tracer). As a model, detection of the pathogen Streptococcus pneumoniae was investigated using a probe against the single-copy lytA gene. RESULTS: Detection of S. pneumoniae (in particular a 1200 p lytA sequences) required less than 1 ng of genomic DNA (approximate size 2.2 Mb). Hybridization and detection were performed in a complex background containing 10 microg fish sperm DNA. CONCLUSIONS: The results indicate the possibility to detect nucleic acid targets in nonamplified DNA samples using easy, inexpensive, amplification free hybridization-based assays and the ultra sensitive UPT reporters. Employment of UPT allows to by-pass target amplification and therefore brings genetic-based testing a step closer to the point-of-care environment. Detection of S. pneumoniae with only 1 ng of DNA indicates a potential for applications in the field of infectious diseases. PMID- 12849875 TI - Plasma homocysteine levels in obese and non-obese subjects with or without hypertension; its relationship with oxidative stress and copper. AB - OBJECTIVES: The relationship between plasma total Homocysteine (tHcy) and oxidative stress and plasma levels of lipids, insulin and copper levels were investigated in obese and nonobese hypertensives. DESIGN AND METHODS: Plasma tHcy levels were determined by an enzyme immunoassay method. Plasma lipid peroxidation levels were measured as thiobarbituric acid reactive substances (TBARS) by spectrophotometric methods. Plasma levels of copper and insulin were measured by atomic absorption spectrophotometer and electrochemiluminescence method, respectively. RESULTS: Plasma tHcy, copper and insulin levels did not differ in nonobese hypertensives compared to nonobese normotensives. Plasma TBARS levels were significantly increased in nonobese hypertensives when compared to nonobese normotensives (p < 0.001). Plasma tHcy, TBARS, copper and fasting insulin levels were significantly higher in obese normotensives and hypertensives than in nonobese normotensives and hypertensives, respectively (for each comparison; p < 0.001). There was a significant difference in plasma tHcy, TBARS and copper levels between obese subjects with or without hypertension (for each comparison p < 0.01). The univariate analyses demonstrated a significant positive correlation between tHcy and TBARS (coefficient +/- SE, 0.411 +/- 0.115, p < 0.01) and copper (coefficient +/- SE, 0.425 +/- 0.135, p < 0.01) in obese subjects. In a multivariate regression analysis in obese subjects tHcy was positively correlated with TBARS (coefficient +/- SE, 0.480 +/- 0.155, p < 0.01) and copper (coefficient +/- SE, 0.486 +/- 0.140, p < 0.01). CONCLUSIONS: We hypothesize that in the presence of other traditional risk factors, Hcy may have a permissive role in the endothelium damage even within the normal range and this role may be related to free radical generating systems. Therefore, modest elevation of plasma Hcy may causally be involved in the pathogenesis of atherosclerosis and/or cardiovascular disease. PMID- 12849877 TI - Magnetoencephalography in pediatric neurology and in epileptic syndromes. AB - In recent years, great advances in the knowledge of neuromagnetism have permitted the application of Superconducting Quantum Interference Devices to the pathophysiologic study of the human brain. In particular, in pediatric neurology, the integration of biomagnetism with magnetic resonance imaging and other techniques for medical imaging have allowed for precise neuromagnetic measurements of the human brain. The more frequently used technique is magnetoencephalography. Recent data have illustrated the usefulness of magnetoencephalography in mapping activity of sensory and motor areas and in studying the spatiotemporal pattern of brain activation specific to somatosensory function. Moreover, magnetoencephalography is an important tool to localize epileptic activity; magnetic source imaging superimposes magnetoencephalographic localizations on the magnetic resonance imaging and yields improved spatial resolution as compared with surface electroencephalography. The role of magnetoencephalography in evaluating patients with epilepsy continues to evolve; in fact, it seems to be very useful in the localization of the epileptogenic zone in patients with partial epilepsy. This application of magnetoencephalography is essential in the selection of epileptic children candidates to surgical treatment of seizures. PMID- 12849878 TI - Long-term outcome in children with gliomas of the anterior visual pathway. AB - We performed a retrospective assessment of the long-term visual, neurologic, and systemic outcomes of 47 patients with anterior visual pathway gliomas seen at the Johns Hopkins Hospital. All of the patients had follow-up of at least 10 years or died during the follow-up period. Two patients died before 10 years of follow-up were achieved. The remaining 45 patients (including three patients who subsequently died) had follow-up of 10-28 years (mean, 15.3 years; median, 15 years). Sixteen of the patients in this study, most of whom had neurofibromatosis type 1 (NF1), received no treatment. None of these patients died or developed neurologic morbidity as a result of their tumor. Thirty-one of the patients, most of whom did not have evidence of NF1, received treatment. Many of these patients subsequently developed neurologic, endocrine, or visual morbidity. However, although patients with anterior visual pathway gliomas who were not treated fared better visually, neurologically, and systemically than patients who were treated, patients who required treatment for progression generally had a good overall prognosis, particularly patients with tumors that did not involve the hypothalamus. Most of these patients survived and maintained useful vision in at least one eye. We believe that patients with anterior visual pathway gliomas, particularly those with NF1, should not be treated unless there is clear clinical or neuroimaging evidence of progression. PMID- 12849879 TI - Diagnostic capability of CSF ferritin in children with meningitis. AB - The current study was performed to determine the normal cerebrospinal fluid (CSF) ferritin level according to age and cut-off value for early diagnosis of bacterial menignitis. The subjects (N = 203) consisted of children who received the CSF examination at Department of Pediatrics in Chonnam National University Hospital between May 1996 and July 2001. The subjects were classified into four groups; non-meningitis, viral meningitis, bacterial meningitis, and bacterial meningitis suspected group. CSF ferritin of the meningitis group was significantly higher than that of the non-meningitis or viral meningitis groups. CSF ferritin had positive correlation with white blood cell (WBC) count and protein in CSF but negative correlation with CSF glucose (P < 0.01). CSF ferritin decreased progressively up to 1 year but such a tendency was not evident in patients over 1 year in age in the non-meningitis group. For early diagnosis of bacterial meningitis, 15.6 ng/mL was considered as the appropriate cut-off value of CSF ferritin (a sensitivity of 96.2% and a specificity of 96.6%). PMID- 12849880 TI - Uncoupling of EEG-clinical neonatal seizures after antiepileptic drug use. AB - A prospective study of the efficacy of seizure cessation by phenobarbital versus phenytoin administration utilized both clinical and electroencephalographic expressions of seizure behaviors. The phenomenon of uncoupling was defined as the persistence of electrographic seizures despite the suppression of >or=50% clinical seizures after either one or both antiepileptic drugs use. Fifty-nine neonates (25 to 43 weeks estimated gestational age) with electrically-confirmed seizures were assigned to either of two drugs and continuously monitored over a 24-hour period. Nine of the fifty-nine patients had only electrographic seizure expression both before and after drug administration. Of the remaining 50 patients who had both electrical and clinical seizure expression before treatment, 24 infants responded to the first choice of an antiepileptic drug with no further seizures. Fifteen of the remaining 26 infants (58%) with persistent seizures after treatment had uncoupling of electrical and clinical expressions of seizures; no difference in the uncoupling effect was noted for neonates who were treated with either antiepileptic drug or based on prematurity or gender. Serial electroencephalographic monitoring helps document continued electrographic seizure expression after antiepileptic drug use, following complete or partial suppression of clinical seizure behaviors. PMID- 12849881 TI - Neurologic outcome of acute measles encephalitis according to the MRI patterns. AB - The aim of this prospective study was to clarify the ramifications of neuroimaging patterns associated with neurologic outcomes of acute measles encephalitis. Twelve patients with neurologic complications stemming from a measles outbreak were diagnosed based on characteristic clinical features, the presence of measles-specific antibodies, and abnormal cerebrospinal fluid findings. Neuroimaging studies were then performed during the acute phase of the disease. Our subject group consisted of four males and eight females (mean age: 9.9 years). Main neurologic findings were loss of consciousness, seizures, and paralysis. Serum tests for measles-specific Ig G and M antibodies were positive in eleven patients. Neuroimages of the group revealed the following three abnormalities: (1). demyelination of white matter with hypoperfusion on interictal single-photon-emission computed tomography, (2). high signal intensity of gray matter with hypoperfusion on interictal single-photon-emission computed tomography, and (3). normal MRI with hypoperfusion on single-photon-emission computed tomography. We identified a new pattern of acute measles encephalitis that involved cortical and/or deep gray matter, and this finding suggests a variant in the pathogenesis of acute measles encephalitis. Patients that demonstrated a gray matter abnormality had a higher rate of complications and required longer periods of hospitalization. PMID- 12849882 TI - The diagnostic role of short duration outpatient V-EEG monitoring in children. AB - Video-electroencephalographic monitoring enables correlation between behavioral and EEG data, however, because it requires prolonged hospitalization, it may be stressful and expensive. This study aimed to assess the benefits and limitations of this procedure in children. We analyzed 39 children classified according to clinical complaints: doubts about epilepsy classification in 23 (Group I); differential diagnosis with nonepileptic events in eight (Group II); and differential diagnosis between cognitive decline and subtle seizures in eight (Group III). Clinical episodes were recorded in 37 patients (94.9%). In Group I, seizure type was reclassified in 11 patients and epileptic syndrome in nine. In two patients a previously unnoticed seizure type was recorded. In Group II, four patients presented epileptic seizures. In Group III, nonconvulsive status was detected in five. Video-electroencephalographic monitoring enabled major modification of therapeutic approach in 21 patients and guided new neuroimaging studies in 10 patients. In conclusion, in patients with frequent seizures, short video-electroencephalographic monitoring allows proper classification of epileptic syndromes, and diagnosis of nonepileptic seizures, promoting introduction of appropriate treatment with a relatively low cost. PMID- 12849883 TI - Elevated levels of measles antibodies in children with autism. AB - Virus-induced autoimmunity may play a causal role in autism. To examine the etiologic link of viruses in this brain disorder, we conducted a serologic study of measles virus, mumps virus, and rubella virus. Viral antibodies were measured by enzyme-linked immunosorbent assay in the serum of autistic children, normal children, and siblings of autistic children. The level of measles antibody, but not mumps or rubella antibodies, was significantly higher in autistic children as compared with normal children (P = 0.003) or siblings of autistic children (P 4 mg glimepiride once daily for 1.5 years. HbA(1c) was reduced from 8.4% at baseline to 7.1% after 4 months and 6.9% after 1 and 1.5 years (median intra individual change from baseline: -1.4, -1.5, and -1.7%, respectively; P<0.0001). Treatment with glimepiride also resulted in significant and stable weight loss relative to baseline, with the exception of patients with a body mass index of <25 kg/m(2). Mean body weight was reduced from 79.8 kg at baseline to 77.9 kg after 4 months, 77.2 kg after 1 year, and 76.9 kg after 1.5 years (mean intra individual change from baseline: -1.9 kg, P<0.0001; -2.9 kg, P<0.05; -3.0 kg, P<0.005, respectively). Therefore, once daily glimepiride provides effective glycaemic control, and may have advantages over other sulphonylureas, because it exhibits weight neutralizing/reducing effects in patients with Type 2 diabetes. PMID- 12849920 TI - Gln-Arg192 polymorphism of paraoxonase 1 is associated with carotid intima-media thickness in patients of type 2 diabetes mellitus of Chinese. AB - Serum paraoxonase (PON) is a high-density lipoprotein-bound enzyme that can prevent oxidation of low-density lipoprotein by hydrolyzing lipid peroxides and thus exert an anti-atherogenic effect. Recent studies have suggested that glutamine(Q isoform)/arginine(R isoform) polymorphism at position 192 of PON(1) gene is associated with macrovascular disease of type 2 diabetes mellitus (T2DM). We re-investigated this relationship using carotid intima-media thickness (IMT) as a surrogate continuous variable for macroangiopathy. The genotype and allele frequency of PON(1) 192 Q/R polymorphism was assayed by polymerase chain reaction restriction fragment length polymorphism in 152 type 2 diabetic patients and 128 healthy subjects from a population of Chinese Han nationality in ChengDu area. The carotid IMT was measured by B-mode ultrasonography in type 2 diabetic patients. No differences were found in PON(1) gene Q/R polymorphism in the type 2 diabetic patients when compared with the control group. The mean carotid IMT in type 2 diabetic subjects with the QQ, QR and RR genotype was 0.65+/-0.27, 0.83+/ 0.27 and 1.05+/-0.32 mm, respectively. One-way ANOVA showed that IMT was significantly greater in the RR subgroup than in both QR and QQ subgroups (P<0.01). Multivariate logistic regression analysis showed R allele to be the main determinant of IMT variability (OR 4.0 95% CI 2.10-7.40 P=0.005). Our data support the view that 192 R allele of PON(1) gene is a risk factor for macrovascular disease of T2DM in Chinese. PMID- 12849921 TI - Prevalence of metabolic syndrome in an urban population: Tehran Lipid and Glucose Study. AB - The aim of the present investigation was to determine the prevalence of the metabolic syndrome among 103,68 of the adults (4,397 men and 5,971 women) aged 20 years and over, participating in the Tehran Lipid and Glucose Study. The metabolic syndrome was defined by the presence of three or more of the following components: abdominal obesity, hypertriglyceridemia, low HDL-C, high blood pressure, and high fasting glucose. The unadjusted prevalence of metabolic syndrome in the study population was 30.1% (CI 95%: 29.2-31.0) and age standardized prevalence was 33.7% (CI 95%: 32.8-34.6). The prevalence increased with age in both sexes. The metabolic syndrome was more commonly seen in women than in men (42% vs. 24%, P<0.001). Low HDL-C was the most common metabolic abnormality in both sexes. Except for high FPG, all abnormalities were more common in women than in men (P<0.001). Most of those with metabolic syndrome had three components of the syndrome (58%), 33% had four, and 9% had five components. This report on the metabolic syndrome from Iran shows a high prevalence of this disorder. Efforts on promoting healthy diets, physical activity, and blood pressure control must be undertaken. PMID- 12849922 TI - Underuse of evidence-based treatment modalities in diabetic patients with non-ST elevation acute coronary syndrome. A prospective nation wide study on acute coronary syndrome (FINACS). AB - This study was designed to evaluate how new treatment guidelines of acute coronary syndrome (ACS) without ST elevation have been implemented in clinical practice especially in diabetic patients. A prospective follow-up was performed on 501 consecutive patients with suspected ACS without ST elevation admitted to nine hospitals in Finland between 15 January and 11 March 2001. The study group included 143 (29%) diabetic patients. Their risk profile was more severe than in non-diabetic patients; ST-depression on admission electrocardiography 57 versus 38%; P<0.0001, elevated troponin levels 66 versus 56%; P<0.05. Six months composite incidence of death, new myocardial infarction (MI), refractory angina or readmission for unstable angina was 39% in diabetic patients and 20% in non diabetic patients (P<0.0001). In spite of this more severe risk profile, glycoprotein (GP) IIb/IIIa receptor antagonists and statins were used with similar frequency in non-diabetic and diabetic patients (15 vs. 19 and 48 vs. 54%, respectively; P=NS for both). In diabetic patients mean delay for in hospital coronary angiography was longer (6.4 vs. 4.2 days, P<0.05) and it was performed less often (32 vs. 45% P<0.05). Our results show that diabetic patients with ACS have higher risk profile and worse outcome than non-diabetic patients. Despite their indisputable benefits in diabetic patients, statins, GP IIb/IIIa receptor antagonists and invasive strategy were underused or often neglected. Further education is needed to change attitudes and to better implement new guidelines into clinical practice. PMID- 12849923 TI - Study on lifestyle-intervention and impaired glucose tolerance Maastricht (SLIM): design and screening results. AB - The study on lifestyle-intervention and impaired glucose tolerance Maastricht (SLIM) is a 3 years randomised clinical trial designed to evaluate the effect of a combined diet and physical activity intervention program on glucose tolerance in a Dutch population at increased risk for developing type 2 diabetes. Here the design of the lifestyle-intervention study is described and results are presented from the preliminary population screening, conducted between March 1999 and June 2000. In total, 2,820 subjects with an increased risk of having disturbances in glucose homeostasis (i.e. age >40 years and BMI>25 kg/m(2) or a family history of diabetes) underwent a first oral glucose tolerance test (OGTT). Abnormal glucose homeostasis was detected in 826 subjects (30.4%): 226 type 2 diabetes (type 2DM, 8.3%), 215 impaired fasting glucose (IFG, 7.9%) and 385 impaired glucose tolerance (IGT, 14.2%). Both increasing age and BMI were strongly related to the prevalence of IGT and diabetes. After a second OGTT, 114 subjects with glucose intolerance and in otherwise good health were eligible for participation in the intervention study (SLIM). The high prevalence of disturbances in glucose homeostasis observed in the preliminary screening underscore the importance of early (lifestyle) interventions in those at risk for developing diabetes. SLIM will address this topic in the Dutch population. PMID- 12849924 TI - Cognitive impairment, physical disability and depressive symptoms in older diabetic patients: the Fremantle Cognition in Diabetes Study. AB - OBJECTIVE: To determine whether the prevalence of dementia, depression and/or disability in older diabetic subjects warrants an active screening approach by diabetes health care workers. PATIENTS AND METHODS: The initial 223 members of a cohort of 529 eligible diabetic subjects, aged 70 years or over, were screened for cognitive impairment (using the Mini-Mental State Examination (MMSE) and Informant Questionnaire for Cognitive Decline in the Elderly (IQCODE)), physical impairments and depressive symptoms. RESULTS: Virtually all subjects were community-dwelling (99%) and mobile (86%) and relatively few had moderate or severe deficits in activities of daily living (ADL) (17.5%). The prevalences of cognitive impairment and probable dementia estimated from the screening tests were high (range 10.8-17.5%) compared with population studies. Any deficit with ADL was reported by 53% of the subjects and common problems included urinary and faecal incontinence. Scores consistent with clinical depression were reported by 14.2% of the sample but 50.2% of the remainder reported one or more depressive symptoms below the cut-off point for clinical depression. Only 36% of the study subjects were free of deficits in any domain. CONCLUSIONS: Community-living older diabetic subjects have high rates of cognitive impairment, deficits in physical function and depressive symptomatology suggesting that screening programs for functional impairments and depression may be beneficial in health care systems that manage older diabetic patients. PMID- 12849925 TI - Prevalence of diabetes, impaired fasting glucose and insulin resistance syndrome in an urban Indian population. AB - OBJECTIVE: Epidemiological study among urban subjects in western India to determine prevalence of diabetes, insulin resistance syndrome (IRS) and their risk factors. METHODS: Randomly selected adults > or =20 years were studied using stratified sampling. Target sample was 1,800 (men 960, women 840). 1123 subjects (response 62.4%) were evaluated and blood samples were available in 532 men and 559 women (n=1091, 60.6%). Measurement of anthropometric variables, blood pressure, fasting blood glucose and lipids was performed. Atherosclerosis risk factors were determined using current guidelines. Diabetes was diagnosed when the subject was a known diabetic or fasting blood glucose was > or =126 mg/dl, impaired fasting glucose (IFG) diagnosed when fasting glucose was 110-125 mg/dl. IRS was diagnosed when any three of-IFG, high triglycerides >150 mg/dl, low HDL cholesterol (men<40 mg/dl, women<50 mg/dl), central obesity (men>102 cm, women>88 cm), or high normal blood pressure (>130/>85 mmHg) or hypertension-were present. RESULTS: Diabetes was present in 70 men (13.2%) and 64 women (11.5%). Age adjusted prevalence of diabetes was 9.3% in men (95% confidence intervals (CI) 6.7-11.8), 8.1% in women (CI 5.8-10.4) and 8.6% overall (CI 6.9-10.3). IFG was in 28 men (5.3%) and 29 women (5.2%). IRS was present in 52 men (9.8%) and 114 women (20.4%) with age-adjusted prevalence of 7.9% in men (CI 6.7-9.1) and 17.5% in women (CI 14.4-20.6) with an overall prevalence of 12.8% (CI 10.8-14.8). Other metabolic abnormalities of IRS in men and women were high triglycerides in 32.1 and 28.6%, low HDL cholesterol in 54.9 and 90.2%; central obesity in 21.8 and 44.0%, and high normal blood pressure or hypertension in 35.5 and 32.4%. IFG subjects had similar atherosclerosis risk factor profile as normal subjects while those with IRS and diabetes had significantly greater prevalence of obesity, central obesity, hypertension, high triglycerides and low HDL (P<0.01). CONCLUSIONS: There is s significant prevalence of diabetes and IRS in this urban Indian population. Subjects with diabetes as well as IRS have greater prevalence of obesity, central obesity, hypertension, hypertriglyceridemia and low HDL as compared with normal subjects. PMID- 12849926 TI - Can human pancreatic beta-cells regenerate in vivo? PMID- 12849929 TI - Involvement of serotonin in depression: evidence from postmortem and imaging studies of serotonin receptors and the serotonin transporter. AB - Definitive conclusions on the role of serotonin receptors and transporter in suicide and depression have been elusive in studies of postmortem brain tissue. A number of methodological differences in these studies have made it difficult to reach a consensus, but crucial issues are being identified and incorporated into newer studies. This review will follow the evolution of serotonin receptor and transporter studies in postmortem tissues that initially focused on suicide and gradually incorporated depressive disorders as psychiatric assessments were increasingly performed. Studies in postmortem tissues on the serotonin-1A and serotonin-2A receptors and the serotonin transporter will be reviewed and compared with imaging studies of the same sites in depressed subjects. Critical issues to control in future studies of serotonin receptors in postmortem tissues include variables such as the cause of death (i.e. suicide), the specific psychiatric diagnoses of the subjects, whether the disorder was in remission at the time of death, long-term medication histories, psychoactive substance use disorders, the smoking history, the hemisphere from which tissues were dissected, and the specific cytoarchitectonic region to be evaluated. Carefully controlled studies in postmortem tissues will ensure a greater likelihood of reaching a consensus on the involvement of monoamine measures in the etiology of depression. PMID- 12849930 TI - Genetics of the serotonergic system in suicidal behavior. AB - Genetic factors contribute to the risk of psychopathology in many psychiatric conditions, but the specific genes are yet to be identified. Neurotransmitter alterations are implicated in the etiology of psychopathology based, in part, on studies of neurotransmitter receptors and their biosynthetic or degradative enzymes in postmortem tissue. Identification of the altered receptors and enzymes serves to identify candidate genes of potential etiological significance. Polymorphisms in these genes can contribute to alterations in protein function in vivo that are part of the neurochemical underpinnings of psychopathologies such as major depressive disorder, psychoses, alcoholism, personality disorders, aggressive-impulsive traits, or suicidal behavior. Altered serotonergic function is implicated in the etiology and pathogenesis of several major psychiatric conditions. In particular, there is much evidence for an association of lower serotonergic function and suicidal behavior. Thus genes related to the serotonergic system are candidate genes worthy of study as part of the genetic diathesis for suicidal behavior. This review examines the following polymorphisms in the serotonin biosynthetic enzyme tryptophan hydroxylase (TPH; A779C substitution), the serotonin transporter (5-HTT, 5-HTTLPR allele), the 5-HT(1B) receptor (G861C, C129T substitution) and the 5-HT(2A) receptor (T102C) for their relationship to suicidal behavior. For the TPH gene, we found the less common U or A allele variant of the A779C polymorphism was associated with suicide attempt. Other studies have found the U allele to be associated with aggression and lower serotonergic function in vivo. A 44 base pair insertion/deletion in the 5' flanking promoter region of the 5-HTT gene may result in less 5-HTT expression and 5-HTT binding. We examined 220 cases postmortem and found no association between the promoter genotype and 5-HTT binding. We also found no association with major depressive disorder (MDD), suicide or pathological aggression, despite finding significantly fewer 5-HTT sites in the prefrontal cortex of depressed and/or suicide cases. In genomic DNA samples from 178 unrelated subjects, we detected two polymorphisms for the 5-HT(1B) receptor at nucleotides 861 and 129. However, no association between either polymorphism and depression, suicide, aggression, or alcoholism was observed. There are two common polymorphisms for the 5-HT(2A) receptor gene in humans. The results of studies of 5-HT(2A) receptor gene polymorphisms do not indicate significant major associations with suicidal behavior. In contrast, the 5-HT(2A) receptor itself is reported to be increased in suicide. Functional polymorphisms involving the promoter region that affect gene expression may explain this finding. Studies of candidate genes related to serotonergic function in brain are increasingly used to establish genetic alterations contributing to psychiatric illness. The most meaningful studies combine the study of candidate genes with direct measures of related proteins as well as psychopathology. PMID- 12849931 TI - Serotonin transporter and MAO-B levels in monoamine nuclei of the human brainstem are normal in major depression. AB - Neurochemical imbalance between noradrenergic and serotonergic systems has been postulated to underlie the pathophysiology of psychiatric illnesses involving mood disorders. The present study was designed to examined the possibility that serotonergic innervation of the locus coeruleus (LC) is abnormal in major depression, by measuring two proteins expressed by serotonergic neurons, but not by noradrenergic neurons, in the region of the LC. The specific binding of [(3)H]paroxetine to serotonin transporter (SERT) and of [(3)H]lazabemide to monoamine oxidase (MAO-B) were measured autoradiographically in tissue sections cut transversely at multiple levels along the rostro-caudal extent of the LC, as well as in the caudal portion of the dorsal raphe nucleus, from psychiatrically normal subjects and age-matched subjects with major depression. Under the conditions of the assays, [(3)H]paroxetine binding in the LC was specific for the SERT, based on the rank order of affinity of compounds for inhibiting [(3)H]paroxetine binding in the LC, i.e. citalopram > imipramine > desipramine > mazindol. The binding of [(3)H]paroxetine to SERT and [(3)H]lazabemide to MAO-B were higher in the raphe nuclei than in the LC. Comparison of control subjects to major depressive subjects revealed no differences in the amount of [(3)H]paroxetine binding to SERT and [(3)H]lazabemide to MAO-B in the LC, as well as in the raphe nuclei. These findings imply that serotonergic innervation of the LC is intact in major depression. PMID- 12849932 TI - Differential expression of alpha2-adrenoceptor vs. imidazoline binding sites in postmortem orbitofrontal cortex and amygdala of depressed subjects. AB - Clonidine is a well established antihypertensive agent that is also used effectively to treat a variety of psychiatric disorders. Clonidine is a prototypic imidazoline compound that acts as an alpha(2)-adrenergic agonist but possesses nearly equivalent affinity for non-adrenergic imidazoline binding sites (I-sites). Receptor autoradiography of [(3)H]-clonidine binding presented herein compares densities of alpha(2)-adrenoceptors and I-sites (under a noradrenergic mask) in Brodmann's area 47 of the left orbitofrontal cortex (OFC) and in six amygdaloid nuclei of subjects with major depression (n=12) vs. controls with no psychiatric history (n=11). Postmortem diagnoses were made from psychiatric interviews with next-of-kin. [(3)H]-Clonidine binding to alpha(2)-adrenoceptors in each of six OFC layers was lower, although not reaching statistical significance in any one layer by multivariate analysis, in depressives vs. control subjects. Binding to I-sites was conversely higher in depressives compared to control OFC layers, but did not reach statistical significance alone. However, the ratios of alpha(2)-adrenoceptor : I-sites in all six layers of OFC of depressed subjects were nearly half that of control subjects (P<0.008). In amygdalas from a different group of depressed patients there were no changes in alpha(2)-adrenoceptors or I-sites, or their ratios, compared with controls. The results support previous western blot data indicating a cortex-selective shift away from alpha(2)AR towards I-site preponderance in depressed patients. PMID- 12849934 TI - Altered expression and phosphorylation of myristoylated alanine-rich C kinase substrate (MARCKS) in postmortem brain of suicide victims with or without depression. AB - Myristoylated alanine-rich C kinase substrate (MARCKS), an acidic, heat-stable protein, is involved in important physiological functions such as neurotransmitter release and re-uptake. It is also a substrate for phosphorylation by protein kinase C (PKC) and has been shown to play a role in the pathophysiology of mood disorders. In this study, protein and mRNA expression of MARCKS as well as phosphorylation of MARCKS were determined in the prefrontal cortex (PFC) and hippocampus of postmortem brain obtained from suicide victims, with or without depression, and normal control subjects. There were no significant differences in mRNA and protein levels of MARCKS between suicide subjects and controls. However, protein levels of MARCKS were significantly increased in the membrane but not in cytosol fraction of PFC and hippocampus obtained from depressed suicide subjects as compared to normal controls. When PKC mediated MARCKS phosphorylation was determined, it was observed that MARCKS phosphorylation was significantly decreased in the membrane fraction of PFC and hippocampus obtained from total suicide subjects as well as depressed and non depressed suicide subjects compared with control population. Although the mechanism of such alterations in MARCKS in depressed and non-depressed suicide subjects is not clear, results of the present study indicate that an increase in membrane MARCKS is associated with depressed suicide victims and a decrease in MARCKS phosphorylation may be a common feature of suicide victims independent of diagnosis. PMID- 12849933 TI - Comparison of prefrontal cell pathology between depression and alcohol dependence. AB - Chronic alcohol abuse is often co-morbid with depression symptoms and in many cases it appears to induce major depressive disorder. Structural and functional neuroimaging has provided evidence supporting some degree of neuropathological convergence of alcoholism and mood disorders. In order to understand the cellular neuropathology of alcohol dependence and mood disorders, postmortem morphometric studies have tested the possibility of alterations in the number and size of cells in the prefrontal cortex and other brain regions. The present review compares the cell pathology in the prefrontal cortex between alcohol dependence and depression, and reveals both similarities and differences. One of the most striking similarities is that, although pathology affects both neuronal and glial cells, effects on glia are more dramatic than on neurons in both alcohol dependence comorbid with depression and idiopathic depression. Moreover, prefrontal cortical regions are commonly affected in both depression and alcoholism. However, the cellular changes are more prominent and spread across cortical layers in alcohol dependent subjects than in subjects with mood disorders, and changes in glial nucleus size are opposite in alcoholism and depression. It could be argued that one defining factor in the manifestation of the depressive pathology is a reduction in the glial distribution in the dlPFC that is reflected in a reduced glial density. In alcoholism reduced glial nuclear size might be related to the cytotoxic effects of prolonged alcohol exposure, while in MDD, in the absence of alcohol abuse, other processes might be responsible for the increase in average size of glial nuclei. In either case abnormal function related to glial reduction would be associated with depression due to insufficient glial support to the surrounding neurons. PMID- 12849935 TI - Lithium inhibitable enzymes in postmortem brain of bipolar patients. AB - Despite considerable ongoing efforts at the epidemiological, genetic and molecular level, the etiology of bipolar disorder had not yet been elucidated. To study possible contributing components to the pathophysiology of this disorder, we have hypothesized that levels of enzymes inhibited by therapeutically relevant lithium ion concentrations in the brain of patients may differ from those in normal controls and may be involved in the etiology of the disorder. Three Li inhibitable enzymes were studied in postmortem brain samples of bipolar patients and normal controls. The expression and function of the two enzymes that are obviously involved in signaling cascades, IMPase, involved in the second messenger system of the phosphatidylinositol cycle, and GSK-3, a mediator of an array of signaling cascades, were not found to be different in postmortem frontal and occipital cortex of bipolar patients and normal controls. Only PAP phosphatase protein levels, but not its mRNA levels or enzymatic activity, were found to be significantly decreased in frontal cortex of bipolar patients compared with normal controls. PMID- 12849936 TI - Assessing outcome after hand surgery: the current state. AB - An overview of the current state of outcome measurement after hand surgery is presented. The paper focuses on the development, application and strengths and weaknesses of it also reviews the existing questionnaires and suggests recommendations for use in research or clinical practice. PMID- 12849937 TI - Outcome of hand trauma: the hand injury severity scoring system (HISS) and subsequent impairment and disability. AB - This study was designed to evaluate correlations between the hand injury severity scoring system (HISS) and measures of impairment and disability obtained 6 months after a hand injury. A statistically significant positive correlation was found between the severity of the injury (HISS) and residual impairment, as measured with the American Medical Association's (AMA) "Guides to the evaluation of permanent impairment, 4th edition". No statistically significant correlation was found between the severity of the injury (HISS) and the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire, which measures disability from the patient's perspective. There was however a statistically significant correlation between AMA total body impairment measured and the results of the DASH questionnaire. The results indicate the potential of the HISS for predicting the final impairment after hand injuries. PMID- 12849938 TI - Local anaesthetic infusion for postoperative pain. AB - The role of continuous bupivacaine infusion either into the wound or as a local nerve block, following hand surgery was investigated in 100 patients. After excluding six patients with complex pain problems in whom neither the bupivacaine infusion nor any other conventional analgesic techniques provided adequate analgesia post-operatively, 86 of 94 (91%) patients were adequately treated for post-operative pain by this system during the first night after surgery when pain is presumed to be greatest. This system also provided adequate on-going analgesia for up to 1 week after surgery, controlling nerve pain and allowing mobilization of tendons after tenolysis. Continuous bupivacaine infusion is of particular use in these two groups of patients and after major hand injuries, when considerable pain can be anticipated. Pain during the first night was not controlled adequately by the bupivacaine infusion system in eight of the 94 patients (8%). All eight had a technical failure of the system, which was rectified in six cases to restore adequate analgesia by the infusion system. Two patients developed infection at the infusion cannula insertion site, which occurred only after 1 week and was successfully treated by removal of the cannula and oral antibiotics. PMID- 12849939 TI - The natural history of an untreated isolated scapholunate interosseus ligament injury. AB - The natural history of an untreated isolated scapholunate interosseus ligament injury remains unclear, although it is commonly assumed that patients continue to suffer with pain, stiffness and weakness of the wrist and ultimately develop secondary osteoarthritis (SLAC wrist). In this study, we evaluated the clinical condition of 11 patients with an arthroscopically proven interosseus scapholunate ligament injury, but without any radiological signs of either DISI deformity or scapholunate gapping, who had declined further treatment at an average follow-up of 7 years. Whilst there was on going pain and functional limitation in all cases, there was no rapid progression to degenerative change (SLAC wrist). PMID- 12849940 TI - Scapholunate fusion in chronic symptomatic scapholunate instability. AB - Since 1989 scapholunate fusion has been performed on 13 patients with chronic scapholunate instability causing debilitating symptoms. These cases were reviewed at a mean 93 (range, 60-132) months after surgery. Establishing whether bony fusion had been achieved proved extremely difficult even after CT scanning, but fusion was unequivocally achieved in four cases. Ten patients were subjectively satisfied with their treatment. Two patients who had no symptomatic improvement subsequently underwent total wrist arthrodesis. A method of fusing the scaphoid and lunate is described, though we accept that a firm fibrous union may be all that is achieved in most patients. However, this appears sufficient to restore stability with a high patient satisfaction. PMID- 12849941 TI - An anatomical database of carpal bone measurements for intercarpal arthrodesis. AB - During limited intercarpal fusion it is sometimes difficult to determine correct screw length and staple size. This is because of overlap, and the shape and orientation of the carpal bones on radiography. One hundred complete sets of dry cadaver carpal bones were measured to provide an anatomical database of carpal bone measurements. This should help prevent inadvertent overpenetration of fixation devices during surgery. PMID- 12849942 TI - The results of treatment of 734 fresh, simple fractures of the scaphoid. PMID- 12849943 TI - Dr. Jorg Bohler discusses his father's work. PMID- 12849944 TI - Trauma-related distress and mood disorders in the early stage of an acute traumatic hand injury. AB - The aim of the study was to estimate the incidence of trauma-related distress and mood disorders in the early stages after acute traumatic hand injuries and identify characteristics associated with these reactions. Data were obtained from 112 patients by means of mailed questionnaires and medical records. Nearly half of the patients had increased levels of intrusive and avoidance symptoms, indicating trauma-related distress. One-third showed signs of a mood disorder. Mood disorders were associated with the need for help with activities of daily living, pain and avoidance symptoms. The study showed that emotional problems in the early stages after injury are related to the consequences of both the injury and the traumatic experience. Negative reactions to the sight of the hand were associated with both trauma-related distress and mood disorders, suggesting that observation of the reactions to the sight of the hand could help to identify patients in need of psychological support. PMID- 12849945 TI - Early parental experiences of obstetric brachial plexus palsy. AB - Early parental experiences of having a child with obstetric brachial plexus palsy were examined to determine whether there were any areas of dissatisfaction and, if so, whether these resulted from their distress and a need to blame someone, or from problems in the communication of bad news. A high level of dissatisfaction was reported with similar levels of dissatisfaction found in a mildly injured group that did not require surgery, and a severely injured group that did. This suggests either that degree of dissatisfaction is not related to degree of disability, or that dissatisfaction was due to factors other than a reaction to their child having a problem. Support was found for the latter as the parents reported details of problems in communication, particularly the giving of inaccurate and misleading information. PMID- 12849946 TI - Biomechanical analysis of a modification of Tang method of tendon repair. AB - We report a modification of the original Tang technique of tendon repair which uses fewer sutures and has fewer knots on the tendon surface. The modified method consists of six longitudinal and two horizontal strands that form an "M" configuration within the tendon and four dorsal longitudinal strands made with a single looped suture. Thirty-six fresh pig flexor tendons were divided and repaired with either the modified Tang or the Tang method. The tendons were subjected to linear or 90 degrees angular loading in an Instron tensile machine. The gap formation strength and ultimate strength of the modified Tang repair was statistically identical to those of the Tang method under linear tension. Under angular tension, the ultimate strength of the modified Tang method was greater than that of the Tang method. PMID- 12849947 TI - The different characteristics of Dupuytren's disease fibroblasts derived from either nodule or cord: expression of alpha-smooth muscle actin and the response to stimulation by TGF-beta1. AB - Mechanisms behind the onset and progression of Dupuytren's disease are poorly understood. Both myofibroblasts and transforming growth factor beta 1 (TGF beta(1)) have been implicated. We studied fibroblast cultures derived from nodules or cords of Dupuytren's contracture tissue to determine the proportion of myofibroblasts present in comparison with flexor retinaculum fibroblast cultures. We identified myofibroblasts by immunohistochemical staining for alpha-SMA. We then investigated the effects of TGF-beta(1) stimulation on these fibroblasts. Basal myofibroblast/fibroblast proportions were 9.7% in nodule cell cultures, 2.7% in cord cell cultures and only 1.3% in flexor retinaculum cell cultures. Nodule and cord myofibroblast proportions increased to 25.4% and 24.2%, respectively, in response to TGF-beta(1) treatment. Flexor retinaculum cell cultures showed no response to TGF-beta(1) stimulation. Fibroblasts cultured from specific regions of Dupuytren's tissue retain myofibroblast features in culture. TGF-beta(1) stimulation causes an increased myofibroblast phenotype to similar levels in both nodule and cord, suggesting that previously quiescent cord fibroblasts can be reactivated to become myofibroblasts by TGF-beta(1). This could be an underlying reason for high recurrence rates seen after surgery or progression following injury. PMID- 12849948 TI - Influence of core suture material and peripheral repair technique on the strength of Kessler flexor tendon repair. AB - The purpose of our study was to determine the most favourable combination of core suture material and peripheral repair technique for Kessler tendon repair. Thirty freshly thawed pig flexor tendons were repaired by a Kessler technique, either with braided polyester or monofilament nylon suture. A peripheral augmentation was done using one of the three techniques-running, cross-stitch and Halsted. All repairs were tested by cyclic loading, followed by load-to-failure. During cyclic loading six of the 15 tendons with a nylon core failed, but none with a braided polyester core. Irrespective of peripheral technique, the monofilament nylon core suture allowed early central cyclic gapping, resulting in failure of the repair. During load-to-failure testing, the running stitch proved weakest and the cross stitch repair toughest. PMID- 12849949 TI - Miniplate fixation for avulsion injuries of the flexor digitorum profundus insertion. AB - This article describes the use of a miniplate and cortical screws in the treatment of five cases of flexor digitorum profundus (FDP) tendon avulsion. One case was type II, three cases were type III and one case was type IV. Near normal joint congruity was restored together with bony union in all cases. Six months after surgery four cases had near normal range of motion at the distal interphalangeal joint compared with the contralateral uninjured finger. These four patients were to return to their previous activities without restriction by 3 months. One repair of a type III avulsion ruptured but the distal interphalangeal joint was pain free and stable and the patient declined further surgery. Miniplate fixation offers some advantages over existing methods of repair and adds to the range of techniques available for reattachment of the FDP tendon in these injuries. PMID- 12849950 TI - A biomechanical study of reattachment versus distal multistrand repair in distal division of the flexor digitorum profundus. AB - Distal division of the flexor digitorum profundus (FDP) within 10mm of its insertion is commonly treated in the same manner as avulsion of the FDP, using the "button" technique or bone suture anchor fixation. Button and bone suture anchor fixation techniques have been associated with significant complications. Importantly, both lead to shortening of the FDP which may cause flexion contracture at the distal interphalangeal joint. This study compared the breaking strength of a multistrand distal suture with reattachment using the "button-on the-nail" technique in a laboratory cadaver model of distal FDP division. The data showed that multistrand distal suture repair was at least as strong as reattachment and has the theoretical advantage of avoiding some of the acknowledged complications of reattachment techniques. PMID- 12849951 TI - The rheumatoid wrist in black South African patients. AB - The wrist and hand X-rays of 75 black patients with rheumatoid arthritis were scored according to the Larsen criteria. The mean Larsen score for left hands was 9.6 (range 0-100) and for the right hands was 10:3 (range 0-100), whereas the score for the wrists were 2.5 (range 0-5) for the left and 2.7 (range 0-5) for the right. Our conclusion is that rheumatoid wrist involvement in black patients was more or less the same as reported in other series, but finger joint involvement was considerably less. This finding must influence surgical decision making and also the interpretation of results of drug trials, whenever black patients are involved. PMID- 12849952 TI - Hand injuries in young children. AB - Four hundred and fifty five young children (0-6 years old) were treated for hand injuries between 1996 and 2000. Boys (61%) were injured more often and a higher number of injuries occurred during May and September. Fingertip injuries were the most common injuries (37%), and were often caused by jamming in doors at home. Fractures were caused by falls and punches and tendon/nerve injuries by sharp objects. The incidence of hand injuries increased from 20.4/10,000/year in 1996 to 45.3/10,000/year in 2000. Only 4% of the children had complex injuries but these placed a high demand on resources. The incidence of injuries was not higher amongst children from immigrant families. PMID- 12849953 TI - Audit of pattern of closures to acute hand services in Pan Thames area. PMID- 12849954 TI - Response to "Hyaluronidase versus surgical excision of ganglia: a prospective randomised clinical trial". PMID- 12849955 TI - Thoughts on "High bifurcation of median nerve at the wrist causing common digital nerve injury in endoscopic carpal tunnel release". PMID- 12849957 TI - Yolk steroid hormones and sex determination in reptiles with TSD. AB - In reptiles with temperature-dependent sex determination (TSD), the temperature at which the eggs are incubated determines the sex of the offspring. The molecular switch responsible for determining sex in these species has not yet been elucidated. We have examined the dynamics of yolk steroid hormones during embryonic development in the snapping turtle, Chelydra serpentina, and the alligator, Alligator mississippiensis, and have found that yolk estradiol (E(2)) responds differentially to incubation temperature in both of these reptiles. Based upon recently reported roles for E(2) in modulation of steroidogenic factor 1, a transcription factor known to be significant in the sex differentiation process, we hypothesize that yolk E(2) is a link between temperature and the gene expression pathway responsible for sex determination and differentiation in at least some of these species. Here we review the evidence that supports our hypothesis. PMID- 12849958 TI - Molecular cloning, characterisation, and tissue distribution of oestrogen receptor alpha in eelpout (Zoarces viviparus). AB - A cDNA encoding the eelpout (Zoarces viviparus) oestrogen receptor alpha (eERalpha) has been isolated from eelpout liver, cloned and sequenced. The cDNA contains a complete open reading frame encoding 570 amino acid residues (mw: 63.0 kDa). The amino acid sequence of eERalpha showed a high degree of identity to ERalpha of other teleost species. The tissue distribution of eERalpha mRNA was examined using Northern blotting, RT-PCR and in situ hybridisation (ISH). All three methods identified a pronounced expression of eERalpha in liver, pituitary, testis and ovary. In the brain ISH experiments showed that ERalpha mRNA was highly expressed in distinct regions of the preoptic area and the mediobasal hypothalamus. We have provided evidence that the receptor is auto-regulated by 17beta-oestradiol (E(2)) not only in liver but also in the testis, indicating an important role for E(2) during spermatogenesis in male eelpout. RT-PCR analysis showed a broader expression pattern including significant expression in the brain, kidney, heart, and gut of adult eelpout. In eelpout embryos eERalpha expression has also been identified, indicating a possible role for the receptor in early development. This study contributes to the accumulating evidence that in fish E(2) is not only involved in the regulation of liver specific proteins, but has a much broader range of targets. PMID- 12849959 TI - FSH and LH-beta subunits in the preoptic nucleus: ontogenic expression in teleost. AB - In the present study we cloned, sequenced, and confirmed the presence of mRNAs of gonadotropins (FSH-beta, LH-beta subunits) from the brain and pituitary of tilapia, Oreochromis niloticus. Further, we examined the spatio-temporal expression pattern of FSH-beta and LH-beta in the brain and pituitary of two species of teleost (tilapia, O. niloticus; sockeye salmon, Oncorhynchus nerka), using in situ hybridization and immunological methods. The expression of FSH and LH immunoreactivity appeared simultaneously in the brain and pituitary (tilapia, 14 days; sockeye, 51 days after fertilization). In the pituitary, FSH mRNA and peptide expressing cells were distinct from LH expressing cells located in the ventral proximal pars distalis. In the brain, FSH and LH immunoreactivity was co localized in cells of the preoptic nucleus parvocellularis, magnocellularis, and gigantocellularis. Fibers immunoreactive to FSH and LH antisera were seen along the forebrain-hypothalamus and in the neurohypophysis of the pituitary. Double label immunofluorescence revealed FSH and LH immunoreactivity co-localized in arginine vasotocin synthesizing preoptic neurons. Our results show that FSH and LH-producing cells have developmental origins in the brain as well as in the pituitary. In addition, we propose that the brain-derived gonadotropins may function as hypophysiotropic hormones that regulate pituitary cells and along with arginine vasotocin could act as neuromodulators of reproductive behaviors. PMID- 12849960 TI - Immunoreactivity for progesterone in the giant Rohde cells of the amphioxus, Branchiostoma belcheri. AB - In higher animals, it is now generally accepted that neurosteroids are steroids that are synthesized in the brain itself, but it remains unclear where, in terms of the phylogeny of chordates, such neurosteroids are first synthesized? We have tried to detect progesterone immunohistochemically in the central nervous system of the amphioxus, Branchiostoma belcheri, an ancient species of chordate. We found immunoreactivity specific for progesterone in the giant neurons known as Rohde cells, at sites that included the perikaryon, in axons and in a thick coarse axon in addition to the gonads. Thus, the present progesterone-like substance appeared to be a phylogenetically ancient and ancestral neurosteroid. Rohde cells are known to be the source of Mauthner cells and to act as interneurons and, therefore, it seems possible that progesterone-like substance might be involved in primitive sigmoid movement, acting as a chemical signal. The presence of progesterone-like substance in amphioxus suggests that animals have evolved by effectively exploiting a rather limited number of active compounds. This report is the first, to our knowledge, to demonstrate that the Rohde cells in the spinal cord of amphioxus have progesterone-like substance, which might be involved in swimming. PMID- 12849961 TI - Evaluating the radiation of the POMC gene in teleosts: characterization of American eel POMC. AB - A distinctive feature of the pituitary hormone precursor, proopiomelanocortin (POMC), is the presence of multiple melanocortin core sequences (HFRW), and one copy of the opioid, beta-endorphin. In the older lineages of ray-finned fish (i.e., orders Acipenseriformes and Semionotiformes), certain extant lobe-finned fish (Australian lungfish and African lungfish), and the tetrapods there are three melanocortin regions in POMC: ACTH/alphaMSH, beta-MSH, and gamma-MSH. However, among the teleosts, the most recent radiation of the ray-finned fishes, the gamma-MSH sequence is absent from the POMC genes of euteleosts like the carp, tilapia, chum salmon, sockeye salmon, and rainbow trout. The objective of this study was to determine whether the gamma-MSH sequence still may be present in the POMC gene of a more basal lineage of the teleosts such as a representative from subdivision Elopomorpha. To this end, a POMC cDNA was cloned and sequenced from the pituitary of the American eel, Anguilla rostrata (order Anguilliformes, family Anguillidae). The open reading frame of the eel POMC cDNA was 648 nucleotides in length and encoded 216 amino acids. As predicted, eel POMC contained the deduced amino acid sequences for beta-endorphin, ACTH/alpha-MSH, and beta-MSH. These end-products displayed primary sequence features that are common to ray-finned fish. Eel POMC lacks a gamma-MSH sequence and a large portion of the joining peptide region. In this regard, the eel POMC gene thus displays features very similar to the POMC genes that have been sequenced from euteleosts. Although it is conceivable that the gamma-MSH sequence may be present in representatives from the other basal extant lineages of teleosts (i.e., subdivisions Osteoglossomorpha or Clupeomorpha), it is also possible that the deletion that resulted in the loss of the gamma-MSH sequence occurred in the ancestral neopterygian that gave rise to the teleosts. In this case, the gamma MSH sequence should be absent in all extant teleosts. PMID- 12849962 TI - Occurrence of beta-endorphin binding sites in the pituitary of the frog Rana esculenta: effect of beta-endorphin on luteinizing hormone secretion. AB - The possible effect of proopiomelanocortin-derived peptide, beta-endorphin on frog gonadotrope cells was investigated. Binding and internalization of beta endorphin to pituitary pars distalis cultured cells were visualized by immunofluorescence and analyzed by means of confocal laser scanning microscopy. Using biotinylated endorphin, the time-course of beta-binding showed that this opioid was internalized through receptor-mediated endocytosis, the mechanism in which actin and clathrin were involved; then, the lysosomal degradation program occurred at later stages. The beta-endorphin binding was well antagonized by Naloxone, the opiate receptor antagonist, and up-regulated since more rapid response was obtained in the previously primed cells. The double immunostaining reaction for beta-endorphin and LH beta-subunit revealed that half the beta endorphin labeled cell population was positively immunostained for LH beta subunit, and beta-endorphin was able to induce an increasing trend of LH secretion in cultured pars distalis cells. Therefore, it seems that beta endorphin acts directly on pituitary pars distalis and influences gonadotropin secretion through the interaction with its own receptor. PMID- 12849963 TI - Differential brain distribution of gonadotropin-releasing hormone receptors in the goldfish. AB - The present study describes the differential distributions in the brain of the two goldfish gonadotropin-releasing hormone (GnRH) receptors, using both immunohistochemistry and in situ hybridization approaches. The goldfish GnRH GfA and GfB receptors are variant forms of the same receptor subtype, although with distinct differences in ligand binding characteristics, and differential distributions in the pituitary and body tissues [Proc. Natl. Acad. Sci. USA 96 (1999) 2526]. The goldfish GnRH GfA receptor was found to be widespread throughout the brain, with neurons showing immunoreactivity in the olfactory bulbs, telencephalon, preoptic region, ventro-basal hypothalamus, thalamus, midbrain, motor neurons of the fifth, seventh, and tenth cranial nerves, reticular formation, cerebellum, and motor zone of the vagal lobes. The tracts in the posterior commissure, optic tectum, and motor zone of the vagal lobes also demonstrated immunoreactivity. While the brain was not systematically surveyed for in situ hybridization, hybridization was found in similar locations in the telencephalon, preoptic region, ventro-basal hypothalamus, cerebellum, and optic tectum. Hybridization was additionally found in the medial hypothalamus. The goldfish GnRH GfB receptor was found to have a more restricted distribution in the brain, with neurons showing immunoreactivity in the telencephalon, preoptic region, and ventro-basal hypothalamus. In situ hybridization demonstrated a somewhat wider distribution of expression of the receptor, with hybridization occurring in the preoptic region, ventro-basal and medial hypothalamus, as well as in the thalamus, epithalamus, and optic tectum. The widespread distribution of GnRH GfA receptor, and in particular its localization in the midbrain tegmentum in the region of the GnRH-II neurons, suggests that this receptor may be involved in the behavioral actions of GnRH peptides in the goldfish. PMID- 12849964 TI - Deiodinase type II and tissue specific mRNA alternative splicing in the Australian lungfish, Neoceratodus forsteri. AB - Deiodinase type II metabolises the prohormone T4 (thyroxine) into the biologically active hormone T3 (3,5,3'-triiodothyronine), at the cellular level in extrathyroidal target tissues. In juvenile lungfish, Neoceratodus forsteri, a typical deiodinase type II is present in most tissues. We have identified the full length of a 1.8 kb deiodinase type II mRNA in liver, and a truncated (1.3 kb) version in brain. Both mRNAs have two in frame UGA codons, but only the liver form has a predicted SECIS structure (form 1) in its 3'-UTR. We also report the presence of additional different length transcripts of deiodinase II mRNA, i.e., 3, 4, and 8 kb, in liver, and 8 kb in kidney, heart, and gill tissues. Expression of the longer (approximately 8 kb) transcript is very low. Real-time PCR confirmed the low expression of transcripts in all tissues, suggested by the Northern blot analysis. PMID- 12849965 TI - 15 alpha-Hydroxytestosterone produced in vitro and in vivo in the sea lamprey, Petromyzon marinus. AB - Prior research has shown that the testes of lampreys are able to synthesize 15 hydroxylated steroid hormones in vitro. Here we show that testes of the sea lamprey Petromyzon marinus L. are able to convert tritiated testosterone into tritiated 15alpha-hydroxytestosterone (15alpha-T) in high yield. The identity of the tritiated 15alpha-T has been confirmed by: co-elution with standard 15alpha-T on high performance liquid chromatography (HPLC); co-elution on thin layer chromatography (TLC); co-elution of acetylated tritiated and standard 15alpha-T on TLC; and strong binding to an antiserum developed against 15alpha-T. The strong reaction between the tritiated 15alpha-T and the antiserum has been used to develop a radioimmunoassay (RIA). The RIA operates over the range of 500-2pg per tube; and can be applied directly to plasma samples. This assay has been used to demonstrate that 15alpha-T is present in blood plasma of the sea lamprey. The concentrations of 15alpha-T in captive lamprey were found to be as follows (pg/ml; mean+/-SEM, n): parasitic stage (reproductively immature), <20, n=7; pre ovulatory females, 156+/-30, n=8; ovulated females, 62+/-9, n=5; pre-spermiating males, 275+/-19, n=8; spermiating males, 216+/-48, n=8. When spermiating male plasma was fractionated on HPLC, immunoreactivity was found exclusively in the expected elution position of 15alpha-T. The biological significance of this steroid has yet to be established. PMID- 12849966 TI - Diurnal and annual changes in serum cortisol concentrations in Indo-Pacific bottlenose dolphins Tursiops aduncus and killer whales Orcinus orca. AB - Until present, fundamental studies on cortisol secretory patterns have not been conducted in cetaceans. The objectives of this study were: (1) to examine diurnal changes in serum cortisol concentrations in Indo-Pacific bottlenose dolphins Tursiops aduncus and killer whales Orcinus orca, (2) to investigate annual cortisol changes in killer whales, and (3) to investigate the relationship between cortisol and sex steroids (testosterone and progesterone) concentrations in killer whales. Diurnal changes in serum cortisol concentrations were investigated at various intervals in the two species. In Indo-Pacific bottlenose dolphins, serum cortisol levels exhibited the same episodic fluctuations for 24 h as did diurnal terrestrial mammals: cortisol levels were lower at 18:00 h and higher in the early morning. In killer whales, cortisol concentrations continued to decrease until 18:00 h, after which they fluctuated, and then increased in the next morning. Annual changes in cortisol levels were investigated by collecting blood samples every two weeks from two male killer whales and a pregnant female one twice per day (during 09:00-10:00 and 16:00-17:00 h) throughout a one-year period. Regarding sera collected during 09:00-10:00 h from the female, cortisol concentrations showed cyclic changes having about 4-month intervals. In males, cortisol showed higher concentrations in winter and lower concentrations during the summer season. There was a negative correlation between cortisol and progesterone levels in the female and a negative correlation was also observed between cortisol and testosterone in male no. 2. In the female and male no. 1, cortisol levels during 09:00-10:00 h were significantly higher than those during 16:00-17:00 h, and their data are considered to support observations regarding diurnal changes in cortisol levels in the two cetacean species. PMID- 12849967 TI - Immunohistochemical localization of inhibin and activin-like proteins in the brain, pituitary gland, and the ovary of thin-lipped grey mullet, Liza ramada (Risso). AB - Inhibin and activin, dimeric protein hormones originally isolated from mammalian gonads, are involved in the regulation of vertebrate reproduction. In the present study, we investigated the localization of inhibin and activin subunits (alpha, beta(A), and beta(B)) immunoreactivity in the brain, pituitary, and ovary of female Liza ramada using immunohistochemistry. We demonstrate that activin beta(A) subunit immunoreactivity is found in most of the brain regions: telencephalon (Tel), midbrain tegmentum (MT), and cerebellum (C). In the pituitary gland, activin beta(A) subunit immunoreactivity is observed in nerve fibers that are in close contact with somatotrophs and gonadotrophs. Strong immunostaining of activin beta(B) subunit is detected in melanotropin (MSH) producing cells in pars intermedia (PI), which are in close contact with gonadotrophs. In the ovary, the cytoplasm as well as follicle cells of previtellogenic oocytes exhibited strong immunoreactivity of activin beta(A). However, the activin beta(A) and beta(B) immunoreactivity is decreased when the oocytes entered the vitellogenic stage (recrudescence). The progress in the accumulation of yolk within oocytoplasm is accompanied by a decrease in the activin beta(A) and beta(B) immunoreactivity and eventually the immunoreactivity disappears completely when the oocytes become fully mature. Interestingly, the vitellogenic oocytes of female L. ramada obtained from fresh water (captivity) contain only weak immunoreactivity of activin beta(A) compared to that of saline water (natural habitat). Moreover, immunoreactivity of activin beta(B) is observed in the phagocytic granulosa cells during resorption of oocytes (atresia) within the ovary of female reared in captivity. The immunostaining of inhibin alpha in the ovary is similar to that of activin beta(B) but the reaction intensity is weaker. However, the mature oocytes show a positive reaction for inhibin alpha. These results provide evidence for the presence of inhibin and activin-like molecules within L. ramada and reveal the distribution of these molecules in L. ramada brain-hypophysial-gonadal system. Our observations add some new informations to our current understanding of activin system in fish and suggest that inhibin and activin may play an important role for L. ramada reproduction in the event of oocyte growth and maturation. PMID- 12849968 TI - Gonadotropin-releasing hormone neuronal development during the sensitive period of temperature sex determination in the pejerrey fish, Odontesthes bonariensis. AB - The development of gonadotropin-releasing hormone (GnRH) neurons was studied in relation to the sensitive period of thermolabile sex determination in the pejerrey Odontesthes bonariensis, an atherinid fish from South America. Fish were raised from hatching at three different temperatures: 17 degrees C (100% females), 24 degrees C (70% females), and 29 degrees C (100% males). Three groups of immunoreactive GnRH (ir-GnRH) neurons were identified at the terminal nerve ganglion (TNG), the midbrain tegmentum (MT), and the preoptic area (POA). Immunoreactive GnRH (ir-GnRH) neurons were identified in the TNG at hatching (day 0) and in the MT at day 3 at all the experimental temperatures. In the POA ir GnRH neurons were identified in the nucleus preopticus periventricularis simultaneously with the first appearance of ir-GnRH fibers in the pituitary on days 11, 14, and 17 for larvae kept at 29, 24, and 17 degrees C, respectively. The number of ir-GnRH neurons in the TNG did not show any statistical difference between temperatures. The number of ir-GnRH neurons in the MT increased in number during the experiment for larvae kept at 17 and 24 degrees C but decreased between days 17 and 31 in larvae kept at 29 degrees C. The number of ir-GnRH neurons in the POA increased during development with a peak at day 28 for all temperatures studied and the magnitude of this peak showed a correlation with incubation temperature. These results reinforce the notion that the hypothalamus pituitary-gonadal axis is active during sex determination in pejerrey suggesting a possible role of the central nervous system and GnRH in this process. It is also suggested that GnRH neurons located in the preoptic area might be the physiological transducers of temperature during the temperature sensitive period in this species. PMID- 12849969 TI - Ontogeny of adenohypophyseal cells in the pituitary of the American shad (Alosa sapidissima). AB - The distribution and ontogeny of adenohypophyseal cells have been studied in the pituitary gland of embryos, larvae, and juveniles of the clupeid American shad (Alosa sapidissima) using immunocytochemical techniques. In juvenile specimens, adenohypophysis was composed of rostral pars distalis (RPD), formed by cavities lined by prolactin (PRL), adrenocorticotropic hormone (ACTH), and gonadotropic hormone (GTH) cells; proximal pars distalis (PPD), containing growth hormone (GH), GTH, and putative thyroid stimulating hormone (TSH) cells; and pars intermedia (PI) with somatolactin (SL) and melanophore stimulating hormone (MSH) cells. At 3 days post-fertilization (3 days pre-hatching) the pituitary of embryos consisted of an oval mass of cells, close to the ventral margin of the diencephalon, divided in rostral and caudal regions. At this time PRL and ACTH cells appeared in the rostral region of the adenohypophysis, while SL cells were observed in the caudal region where MSH cells showed reactivity 1 day before hatching. At variance, GH cells showed a weak immunoreactivity in the rostral portion at hatching that increased 2 days latter. GTH cells also showed weak immunoreactivity in the rostral region of the adenohypophysis at hatching time. Two days later GTH cells were located in the rostral and central regions of the adenohypophysis. At hatching, the neurohypophysis was very small and no nerve processes were seen to penetrate the adenohypophysis tissue. After hatching, the pituitary gland elongated and in 7 days old larvae, the RPD showed a small lumen surrounded by a palisade of PRL, ACTH, and GHT cells; the PPD showed GH and GTH cells while the PI contained SL and MSH cells. The adenohypophysis and neural lobe increased in size with development and, in 42 days old larvae, they were similar to those of juvenile specimens. PMID- 12849970 TI - Cloning of complimentary deoxyribonucleic acid encoding follicle-stimulating hormone and luteinizing hormone beta subunit precursor molecules in Reeves's turtle (Geoclemys reevesii) and Japanese grass lizard (Takydromus tachydromoides). AB - Reptilia is the only vertebrate class in which cDNA for the gonadotropin beta subunit precursor molecule has not been cloned. We have isolated the full-length cDNA clone encoding the LH beta subunit precursor molecule and a partial cDNA clone encoding the FSH beta subunit precursor molecule from a pituitary cDNA library of Reeves's turtle. We further clarified the nucleotide sequence of the remaining part of the turtle FSH beta cDNA and that of full-length cDNA encoding the LH beta subunit precursor molecule of the Japanese grass lizard, by means of the 5' rapid amplification of cDNA end (RACE) and 3' RACE. The nucleotide sequence of the turtle FSH beta cDNA we determined was 584 bp long and contained the coding sequence, 5' untranslated region (UTR) and 3' UTR of 396, 34, and 154 bp, respectively. The nucleotide sequence of the turtle LH beta we isolated was 498 bp long and contained the coding sequence, 5' UTR and 3' UTR of 420, 7, and 71 bp, respectively. The nucleotide sequence of the lizard LH beta we determined was 537 bp long and contained the coding sequence, 5' UTR and 3' UTR of 441, 35, and 61 bp, respectively. Amino acid sequences deduced from coding regions of the turtle FSH beta, LH beta and the lizard LH beta were 131, 139, and 146 residues, respectively. Referring to the amino acid sequences of the bullfrog FSH and LH beta subunit molecules determined chemically, we deduced the amino acid sequences of mature peptide. Amino acid sequences of mature peptides of the turtle FSH, turtle LH, and the lizard LH were 111, 112, and 112 residues, respectively. Amino acid sequences of the mature peptides were compared with those of other vertebrates. The amino acid sequence of the turtle FSH beta subunit molecule was 84.7-85.6, 67.8-71.4, and 61.3-62.2% identical to the FSH sequence of birds, mammals, and amphibians, respectively. The amino acid sequence of the turtle LH beta subunit molecule was 51.6-54.6, 36.2-48.7, and 56.3-57.5% identical to the LH sequence of birds, mammals, and amphibians, respectively. The amino acid sequence of the lizard LH beta subunit molecule was 39.1-47.1, 32.9-43.0, and 46.0-47.3% identical to the LH sequence of birds, mammals, and amphibians, respectively. These identity values suggest that the turtle or reptilian FSH beta subunit molecule is more closely related to avian and mammalian FSH beta subunit molecules than to amphibian FSH beta subunit molecules but reptilian LH beta subunit molecules are more closely related to amphibian LH beta subunit molecules than to avian and mammalian LH beta subunit molecules. This discrepancy in the molecular similarity relationship found in the reptilian FSH and LH beta subunit molecules can be interpreted by assuming that evolution speed was not the same among hormone species and also among vertebrate groups. PMID- 12849971 TI - Molecular cloning and characterization of chicken orphan nuclear receptor cTR2. AB - Orphan nuclear receptors belong to the nuclear receptor superfamily of liganded transcription factors, whose ligands either do not exist or remain to be identified. We report here the cloning and characterization of the chicken orphan nuclear receptor, cTR2 (chicken testicular receptor 2). The cTR2 gene encodes a protein of 569 amino acids which shows approximately 72% overall identity with TR2 (NR2C1) and 95% identity in the DNA-binding domain (DBD). The cTR2 gene is expressed in almost all adult tissues and embryonic stages examined unlike its mammalian relative TR2, which is specifically expressed in testis. Electrophoretic mobility shift assays demonstrate that cTR2 binds the canonical direct repeat DNA recognition sequences spaced by one, four, and five nucleotides (DR1, DR4, and DR5), and in consistence with the results with canonical DNA binding sequences, cTR2 forms specific DNA-protein complex with chicken phenobarbital response elements containing DR4 motifs. Both in vitro and in vivo interaction studies demonstrate that cTR2 forms homodimer. Moreover, transient transfection studies reveal its capability to transactivate canonical DR1, DR4, and DR5 sequences and the constitutive activity of cTR2 is mapped to the N terminal region of this orphan receptor. Finally, cTR2 represses transactivation of estrogen receptor in a dose-dependent manner. PMID- 12849972 TI - Molecular cloning of an anuran V(2) type [Arg(8)] vasotocin receptor and mesotocin receptor: functional characterization and tissue expression in the Japanese tree frog (Hyla japonica). AB - In most amphibians, [Arg(8)] vasotocin (VT) has an antidiuretic effect that is coupled to the activation of adenylate cyclase. In contrast, mesotocin (MT) has a diuretic effect and acts via the inositol phosphate/calcium signaling pathway in amphibians. To further clarify the mechanisms of VT and MT activation, we report the molecular cloning of a VT receptor (VTR) and a MT receptor (MTR) from the Japanese tree frog, Hyla japonica. Tree frog VTR or MTR cDNA encoded 363 or 389 amino acids, and their amino acid sequences revealed close similarity to the mammalian vasopressin V(2) (51-52% identity) or toad MT (94% identity) receptors, respectively. Using CHO-K1 cells transfected with tree frog VTR, we observed elevated concentrations of intracellular cAMP following exposure of the cells to VT or other neurohypophysial hormones, whereas the cells transfected with MTR did not exhibit altered cAMP concentrations. The cells transfected with VTR exhibited the following efficiency for cAMP accumulation: VT = hydrin 1 > or = vasopressin > or = hydrin 2 > MT = oxytocin > isotocin. VTR or MTR mRNA exhibits a single 2.2 or 5.5-kb transcription band, respectively, and both are expressed in various tissues. VTR mRNA is clearly expressed in brain, heart, kidney, pelvic patch of skin, and urinary bladder, whereas brain, fat body, heart, kidney, and urinary bladder express MTR mRNA. Specifically, VTR mRNA in the pelvic patch or MTR mRNA in the dorsal skin is present at elevated levels in the skin. Characteristic distribution of VTR and MTR on osmoregulating organs indicates the ligands for these receptors would mediate a variety of functions. Further, the distribution of VTR in the skin would make the regional difference on cutaneous water absorption in response to VT in the Japanese tree frog. PMID- 12849973 TI - A peptide sequence-YSGVCHTDLHAWHGDWPLPVK [40-60]-in yeast alcohol dehydrogenase prevents the aggregation of denatured substrate proteins. AB - The structural and functional characteristics of a yeast alcohol dehydrogenase (ADH) peptide (YSGVCHTDLHAWHGDWPLPVK, residues 40-60) have been studied in detail. The peptide is hydrophobic in nature, binds the hydrophobic probe bis ANS, and is mostly present in a random coil conformation. It shows chaperone-like activity by preventing dithiothreitol (DTT)-induced aggregation of insulin at 27 degrees C, oxidation-induced aggregation of gamma-crystallin at 37 degrees C, and aggregation of thermally denatured ADH and beta(L)-crystallins at 52 degrees C. However, the ADH peptide does not solubilize protein aggregates as do surfactants. Substitution of Pro for His in the ADH peptide leads to diminished anti-aggregation activity. Further, analysis of ADH incubated at 47 degrees C suggests that a significant portion of the enzyme remains as soluble inactive protein with negligible conformational change. Therefore, we propose that the residues 40-60 in native protein may be an intramolecular chaperone site of yeast ADH. PMID- 12849974 TI - Prostate targeting ligands based on N-acetylated alpha-linked acidic dipeptidase. AB - To identify inhibitors of the intrinsic N-acetylated alpha-linked acidic dipeptidase (NAALADase) activity of prostate specific membrane antigen (PSMA) that may be useful for targeting imaging agents or chemotherapeutic drugs to disseminated prostate cancer, analogs of the tetrahedral transition state for hydrolysis of the natural substrate, N-acetylaspartylglutamate (NAAG), were synthesized. These compounds were assayed for their ability to inhibit the membrane-associated enzyme isolated from LNCaP prostate cancer cells. Active inhibitors were further assayed for their cytotoxicity and membrane binding. We have identified nine compounds, including fluorescent and iodine-labeled conjugates, which inhibit NAALADase enzyme activity with IC(50)s at, or below, 120nM. The binding of these compounds to the cell surface of viable LNCaP prostate tumor cells appears to be specific and saturable, and none of the compounds alter the cell cycle kinetics or induce apoptosis in LNCaP cells, suggesting that they are relatively innocuous and are suitable for targeting imaging agents or cytotoxic drugs to disseminated prostate cancer. PMID- 12849975 TI - An amino acid substitution on the second acetylcholinesterase in the pirimicarb resistant strains of the peach potato aphid, Myzus persicae. AB - cDNAs encoding two acetylcholinesterases (AChEs) were isolated from the peach potato aphid, Myzus persicae. MpAChE1 was orthologous and MpAChE2 was paralogous with the ace of Drosophila melanogaster. The deduced amino acid sequence of MpAChE1 cDNA was identical between the pirimicarb susceptible and resistant strains. However, a single amino acid substitution of Ser431Phe on MpAchE2 was found in the pirimicarb resistant strains. This substitution was located in the acyl pocket of the enzyme and was thought to alter the ligand specificity. PMID- 12849976 TI - The bisphosphonate ibandronate stimulates reverse cholesterol transport out of monocytoid cells by enhanced ABCA1 transcription. AB - Nitrogen-containing bisphosphonates used in osteoporosis act by interference with pyrophosphorylated intermediates of the sterol pathway and are internalized by monocytes/macrophages, key players in atherogenesis. We therefore studied the effects of ibandronate on monocytic cholesterol homeostasis. In differentiated human MM6 cells and freshly prepared human PBMCs lipoprotein receptor transcription was quantified by real-time RT-PCR and receptor-mediated cellular cholesterol handling by lipoprotein-driven uptake and efflux assays. Low nanomolar concentrations of ibandronate reduced cellular cholesterol content despite reactive up-regulation of the LDL receptor. Simultaneously, the transcription of the cellular cholesterol exporter ABCA1 was severalfold stimulated, whereas the scavenger receptor CD36 was down-regulated. Thereby, ibandronate decreased the cellular uptake of modified LDL and enhanced the efflux of cholesterol to delipidated HDL. Geranylgeraniol antagonized the stimulation of ABCA1 expression by ibandronate. Ibandronate in low pharmacologic concentrations redirects monocytic cholesterol handling from favouring foam cell formation towards enhanced reverse cholesterol transport. PMID- 12849977 TI - The protein kinase 60S is a free catalytic CK2alpha' subunit and forms an inactive complex with superoxide dismutase SOD1. AB - The 60S ribosomes from Saccharomyces cerevisiae contain a set of acidic P proteins playing an important role in the ribosome function. Reversible phosphorylation of those proteins is a mechanism regulating translational activity of ribosomes. The key role in regulation of this process is played by specific, second messenger-independent protein kinases. The PK60S kinase was one of the enzymes phosphorylating P-proteins. The enzyme has been purified from yeast and characterised. Pure enzyme has properties similar to those reported for casein kinase type 2. Peptide mass fingerprinting (PMF) has identified the PK60S as a catalytic alpha(') subunit of casein kinase type 2 (CK2alpha(')). Protein kinase activity is inhibited by SOD1 and by highly specific CK2 inhibitor-4,5,6,7 tetrabromo-benzotriazole (TBBt). The possible mechanism of regulation of CK2alpha(') activity in stress conditions, by superoxide dismutase in regulation of 80S-ribosome activity, is discussed. PMID- 12849978 TI - Identification of a novel human uridine phosphorylase. AB - Uridine phosphorylase catalyzes the reversible phosphorylytic cleavage of uridine and deoxyuridine to uracil and ribose- or deoxyribose-1-phosphate. The enzyme has an important role in the metabolism of pyrimidine analogs used in cancer chemotherapy. The cDNA of a novel 317 amino acid human uridine phosphorylase approximately 60% identical to the previously identified human uridine phosphorylase was cloned. The novel enzyme, named uridine phosphorylase-2 (UPase 2), showed broad substrate specificity and accepted uridine, deoxyuridine, and thymidine as well as the two pyrimidine nucleoside analogs 5-fluorouridine and 5 fluoro-2(')-deoxyuridine. The human UPase-2 gene was mapped to chromosome 2q24.1 and the 2.2-kb mRNA was predominantly expressed in kidney. The mouse UPase-2 cDNA was also identified and shown to be predominantly expressed in liver. The identification of a novel uridine phosphorylase with broad substrate specificity is important for studies on both nucleoside metabolism as well as for studies on the pharmacological mechanisms of therapeutic pyrimidine nucleoside analogs. PMID- 12849979 TI - Phospholipase C-dependent Ca2+ release by worm and mammal sperm factors. AB - Egg activation in all animals evidently requires the synthesis of inositol 1,4,5 trisphosphate (InsP(3)) from phosphatidylinositol 4,5-bisphosphate (PIP(2)) by phospholipase C (PLC). Depending on the organism, InsP(3) elicits either calcium oscillations or a single wave, which in turn initiates development. A soluble component in boar sperm that activates mammalian eggs has been suggested to be a PLC isoform. We tested this hypothesis in vitro using egg microsomes of Chaetopterus. Boar sperm factor elicited Ca(2+) release from the microsomes by an InsP(3)-dependent mechanism. The PLC inhibitor U-73122, but not its inactive analog U-73343, blocked the response to sperm factor but not to InsP(3). U-73122 also inhibited the activation of fertilized and parthenogenetic eggs. Chaetopterus sperm also contained a similar activity. These results strongly support the hypothesis that sperm PLCs are ubiquitous mediators of egg activation at fertilization. PMID- 12849980 TI - Promoter hypomethylation of a novel cancer/testis antigen gene CAGE is correlated with its aberrant expression and is seen in premalignant stage of gastric carcinoma. AB - Previously, we reported the identification and characterization of a novel cancer/testis antigen gene, CAGE(4), that was expressed in various histological types of tumors, but not in normal tissues, with the exception of the testis. To date, molecular mechanisms for the expression of CAGE have never been studied. In our expression analysis, we found that some cancer cell lines did not express CAGE. The expression of CAGE could be restored in these cell lines by treatment with 5(')-aza-2(')-deoxycytidine, suggesting that the expression of CAGE is mainly suppressed by hypermethylation. Bisulfite sequencing analysis of the 16 CpG sites of the CAGE promoter in various cancer cell lines and tissues revealed a close relationship between the methylation status of the CAGE promoter and the expression of CAGE. The transient transfection experiments displayed that the methylation of CpG sites inhibited the CAGE promoter activity in luciferase reporter assays. The methylation of the CpG sites inhibited the binding of transcription factors, shown by a mobility shift assay. A methylation-specific PCR analysis revealed that hypomethylation of the CAGE promoter was present at frequencies of more than 60% in breast, gastric, and lung cancers, and hepatocellular carcinomas, and at frequencies of less than 40% in prostate, uterine cervical, and laryngeal cancers. Promoter hypomethylation was found in chronic gastritis (19/55, 34.5%) and liver cirrhosis (13/22, 59%), but not in normal prostate, normal colon, or chronic hepatitis. These results suggest that the methylation status of the CpG sites of CAGE determines its expression, that the hypomethylation of CAGE precedes the development of gastric cancer and hepatocellular carcinoma, and that the high frequencies of hypomethylation of CAGE, in various cancers would be valuable as a cancer diagnostic marker. PMID- 12849981 TI - Zinc inhibits calcineurin activity in vitro by competing with nickel. AB - Calcineurin (CN) is a Ca(2+)/calmodulin (CaM)-dependent protein serine/threonine phosphatase that contains Zn(2+) in its catalytic domain and can be stimulated by divalent ions such as Mn(2+) and Ni(2+). In this study, the role of exogenous Zn(2+) in the regulation of CN activity and its relevance to the role of Ni(2+) was investigated. Zn(2+) at a concentration range of 10nM-10 micro M inhibited Ni(2+)-stimulated CN-activity in vitro in a dose-dependent manner and approximately 50% inhibition was attained with 0.25 micro M Zn(2+). Kinetic analysis showed that Zn(2+) inhibited the activity of CN by competing with Ni(2+). Interaction of CN and CaM was not inhibited with Zn(2+) at 10 micro M. Zn(2+) never affected the activity of cAMP phosphodiesterase 1 or myosin light chain kinase (CaM-dependent enzymes) and rather activated alkaline phosphatase. The present results indicate that Zn(2+) should be a potent inhibitor for CN activity although this ion is essential for CN. PMID- 12849982 TI - Amphiphilic properties of (-)-epicatechin and their significance for protection of cells against peroxynitrite. AB - The dietary flavanol (-)-epicatechin protects against nitration and oxidation reactions of the inflammatory mediator peroxynitrite in hydrophilic and hydrophobic environments. Bioavailability and cellular uptake of (-)-epicatechin are not yet fully characterized. Here, the octanol/buffer partition coefficient of (-)-epicatechin is observed to be 1.5, indicating that the flavanol is soluble in aqueous as well as lipophilic cellular phases, thus capable of permeating the cell membrane. In line with this, the ability of murine aortic endothelial cells (MAECs) to remove (-)-epicatechin from cell culture media is demonstrated. Epicatechin accumulates in cells, likely due to epicatechin binding to cellular proteins. Even after repeated washing, (-)-epicatechin accumulated by MAEC affords protection of the cells against peroxynitrite-induced nitration of protein tyrosyl residues and against oxidation of intracellular dichlorodihydrofluorescein. PMID- 12849983 TI - Production of human skeletal alpha-actin proteins by the baculovirus expression system. AB - Mutations within the human skeletal muscle alpha-actin gene cause three different skeletal muscle diseases. Functional studies of the mutant proteins are necessary to better understand the pathogenesis of these diseases, however, no satisfactory system for the expression of mutant muscle actin proteins has been available. We investigated the baculovirus expression vector system (BEVS) for the abundant production of both normal and mutant skeletal muscle alpha-actin. We show that non-mutated actin produced in the BEVS behaves similarly to native actin, as shown by DNase I affinity purification, Western blotting, and consecutive cycles of polymerisation and depolymerisation. Additionally, we demonstrate the production of mutant actin proteins in the BEVS, without detriment to the insect cells in which they are expressed. The BEVS therefore is the method of choice for studying mutant actin proteins causing human diseases. PMID- 12849984 TI - A novel function of connexin 32: marked enhancement of liver function in a hepatoma cell line. AB - Connexin 32 (Cx32) is the main gap junction protein in hepatocytes and plays an important role in the regulation of signal transfer and growth control in the liver by constructing gap junction channels and gap junctional intercellular communication (GJIC). In this study, the human Cx32 gene was transfected into a hepatoma cell line (HepG2) that showed aberrant expression of Cx32 and was deficient in GJIC. Cx32-transfected HepG2 not only expressed a higher level of Cx32 mRNA, but also showed increased GJIC compared with HepG2 and vector transfected HepG2. Furthermore, the liver functions of ammonia removal and albumin secretion of HepG2 were markedly enhanced with Cx32 gene transfection. It may be expected to improve the cellular functions of the hepatoma cell line by Cx32 gene transfection and serve to develop an efficacious bioartificial liver. PMID- 12849985 TI - The protein of a new gene, Tctex4, interacts with protein kinase CK2beta subunit and is highly expressed in mouse testis. AB - Casein kinase 2 (CK2) is a ubiquitous, multifunctional eukaryotic serine/threonine kinase that phosphorylates an array of proteins. CK2 is a heterotetramer composed of two catalytic (alpha,alpha(')) and two regulatory (beta) subunits. CK2 plays an essential role in regulatory pathways in cell transformation and proliferation. But the role and function of the individual subunits of CK2, which are not in the holoenzyme, are not yet clear. Northern blot analysis reveals the highest CK2beta activity in mouse testicles and brain. By employing a yeast two-hybrid screen to identify the proteins that interact with CK2beta, we have isolated a cDNA clone encoding a 14-kDa protein with homology to dynein light chains and have designated it as Tctex4. CK2beta interacts specifically with Tctex4 both in a yeast two-hybrid system and in an in vitro interaction assay. Northern blot and in situ hybridization showed that Tctex4 is a novel gene that is expressed in mouse testis. PMID- 12849986 TI - Mutations within a conserved protein kinase A recognition sequence confer temperature-sensitive and partially defective activities onto mouse c-Rel. AB - We have created two mutants of mouse transcription factor c-Rel (c-G29E and c R266H) that are analogous to mutants previously shown to have temperature sensitive (ts) functions for the homologous Drosophila protein Dorsal and the retroviral oncoprotein v-Rel. In vitro, c-R266H shows both a ts and a concentration-dependent ability to bind DNA, suggesting that the lesion affects the ability of c-Rel to form homodimers. In contrast, the ability of mouse c-G29E to bind DNA in vitro is not ts. c-Rel mutant c-R266H also shows a ts ability to activate transcription from a kappaB-site reporter plasmid, whereas c-G29E activates transcription well above control levels at both 33 and 39 degrees C. Insertion of two amino acids (Pro-Trp) between amino acids 266 and 267 in mouse c Rel (mutant c-SPW) also creates a c-Rel protein with distinct properties: mutant c-SPW is partially defective in that it cannot form DNA-binding homodimers but can form DNA-binding heterodimers with p50. Interestingly, the mutations in c-Rel that affect homodimer formation (c-R266H and c-SPW) fall within a consensus protein kinase A recognition sequence but are not predicted to lie in the dimer interface. Conditional and partially defective mutants such as those described herein may be useful for identifying physiological responses and genes regulated by specific Rel/NF-kappaB family members. PMID- 12849987 TI - Uptake of analogs of penetratin, Tat(48-60) and oligoarginine in live cells. AB - Cell-penetrating peptides are regarded as promising vectors for intracellular delivery of large, hydrophilic molecules, but their mechanism of uptake is poorly understood. Since it has now been demonstrated that the use of cell fixation leads to artifacts in microscopy studies on the cellular uptake of such peptides, much of what has been considered as established facts must be reinvestigated using live (unfixed) cells. In this work, the uptake of analogs of penetratin, Tat(48-60), and heptaarginine in two different cell lines was studied by confocal laser scanning microscopy. For penetratin, an apparently endocytotic uptake was observed, in disagreement with previous studies on fixed cells found in the literature. Substitution of the two tryptophan residues, earlier reported to be essential for cellular uptake, did not alter the uptake characteristics. A heptaarginine peptide, with a tryptophan residue added in the C-terminus, was found to be internalized by cells via an energy-independent, non-endocytotic pathway. Finally, a crucial role for arginine residues in penetratin and Tat(48 60) was demonstrated. PMID- 12849988 TI - Mitochondrial localization of Smad5 in a human chondrogenic cell line. AB - Bone morphogenetic proteins (BMPs) are members of the transforming growth factor beta (TGF-beta) superfamily and regulate the formation of cartilage and bone tissues as well as other key events during development. TGF-beta superfamily signaling is mediated intracellularly by Smad proteins, some of which can translocate into the cell nucleus and influence gene expression. Although much progress has been made in understanding how TGF-beta superfamily signaling regulates expression of target genes, little formal proof has been presented regarding the intracellular distribution of the Smad proteins before their entry into the nucleus. In the literature, non-nuclear Smad proteins are generally referred to as cytoplasmic. Using confocal microscopy, we here show for the first time that immunofluorescent labeling of Smad5, one of the Smad proteins associated with BMP signaling, colocalizes with the mitochondrion-specific probe MitoTracker, demonstrating a mitochondrial distribution of Smad5 in non stimulated chondroprogenitor cells. PMID- 12849989 TI - A chloride-permeable channel from Phaseolus vulgaris roots incorporated into planar lipid bilayers. AB - Ion channels are key participants in physiological processes of plant cells. Here, we report the first characterization of a high conductance, Cl(-)-permeable channel, present in enriched fractions of plasma membranes of bean root cells. The Cl(-) channel was incorporated into planar lipid bilayers and its activity was recorded under voltage clamp conditions. The channel is voltage-dependent, excludes the passage of cations (K(+), Na(+), and Ca(2+)), and is inhibited by micromolar concentrations of Zn(2+). The Cl(-) conductance here characterized represents a previously undescribed channel of plant cells. PMID- 12849990 TI - Regulation of the cAMP-elevating effects of isoproterenol and forskolin in cardiac myocytes by treatments that cause increases in cAMP. AB - Regulation of the cAMP-elevating effects of isoproterenol and forskolin in cardiac myocytes by treatments that cause increases in cAMP. We have found that elevations in cyclic AMP (cAMP) have long-term effects on both the beta adrenergic receptor and adenylyl cyclase in cultured chick ventricular myocytes. Pretreatment with isoproterenol for 15min markedly reduced the cAMP-elevating effect of a subsequent treatment with isoproterenol 18h later. Responses to isoproterenol were similarly reduced after overnight treatments with forskolin or phosphodiesterase inhibitors. Furthermore, these same treatments also markedly blunted the cAMP-elevating effect of forskolin, a direct activator of adenylyl cyclase. The blunting of the isoproterenol effect was greater than that of the forskolin effect, at least partially because the pretreatments caused a decrease in the number of beta-adrenergic receptors as well as a net decrease in adenylyl cyclase activity. Experiments with a recombinant adenovirus to express luciferase under the control of cAMP responsive elements (CREs) showed that the same treatments elevated cAMP sufficiently to drive the transcription of a gene with CREs in its promoter. The blunting of both the isoproterenol and forskolin responses was blocked by the inhibition of protein synthesis or by infecting cells with a recombinant adenovirus that expresses rabbit muscle cAMP-dependent protein kinase inhibitor (PKI). It is hypothesized that one or more adenylyl cyclase isozymes responsible for the generation of cAMP in the myocytes, along with other proteins previously reported to regulate beta-adrenergic receptors and perhaps adenylyl cyclase, are negatively regulated by cAMP, most likely at the level of gene expression, and that this regulation may have therapeutic consequences in the treatment of cardiac diseases. PMID- 12849991 TI - RNA interference-mediated reduction in GLUT1 inhibits serum-induced glucose transport in primary human skeletal muscle cells. AB - Using RNA interference (RNAi), we specifically down-regulate protein expression in differentiated human skeletal myotube cultures. Serum stimulation of myotubes increases glucose uptake. Using a sensitive photolabeling technique, we demonstrate that this increase in glucose uptake is accompanied by increased cell surface content of glucose transporter (GLUT) 1. Using RNAi, we specifically reduce GLUT1 mRNA and protein expression, leading to inhibition of serum-mediated increase in glucose transport. Thus, we demonstrate the utility of RNAi in a primary human differentiated cell system, and apply this methodology to demonstrate that serum-mediated increase in glucose transport in human skeletal muscle cells is dependent on GLUT1. PMID- 12849992 TI - Biopanning of endotoxin-specific phage displayed peptides. AB - Systemic bacterial infections frequently lead to a plethora of symptoms termed "endotoxic shock" or "sepsis." Characterized by hypotension, coagulation abnormalities, and multiple organ failure, treatment of sepsis still remains mostly supportive. Of the various experimental therapeutic interventional strategies, neutralization of endotoxin by peptides or proteins is becoming popular recently. Hence, design of endotoxin binding peptides is gaining currency as their structural complexity and mode of recognition of endotoxin precludes mounting of resistance against them by the susceptible bacteria by genetic recombination, mutation, etc. Earlier work from our laboratory had shown that the amphiphilic cationic peptides are good ligands for endotoxin binding. In this study, we report the results of studies with the 12 selected lipid A binding phage displayed peptides by biopanning of a repertoire of a random pentadecapeptide library displayed on the filamentous M-13 phage. A comparison of the sequences revealed no consensus sequence between the 12 selected peptides suggesting that the lipid A binding motif is not sequence specific which is in accord with the sequence variation seen with the naturally occurring anti microbial and/or endotoxin binding peptides. Thus, the flexibility of the peptides coupled with their plasticity in recognizing the lipid A moiety, explains their tight binding to endotoxin. At a structural level, asymmetric distribution of the charged polar residues on one face of the helix and non-polar residues on the opposite face appears to correlate with their activity. PMID- 12849993 TI - Hormonal regulation and characterisation of the mouse Cyp4b1 gene 5'-flanking region. AB - The Cyp4b1 structural gene comprises 12 exons that span approximately 23kb. The transcriptional initiation site, mapped by 5(')-RACE and primer extension analyses from kidney mRNA, was found to be 34-bp upstream of the translation initiation site. Cyp4b1 mRNA expression was found restricted to kidney and androgenic up-regulation in several mouse strains was observed. Spatial location investigated by in situ hybridisation revealed abundant and specific expression of Cyp4b1 mRNA in the proximal tubules of the renal cortex. A 1.8-kb murine Cyp4b1 5(')-flanking region, which encompasses a TATA box-like sequence and several putative transcription factor-binding sites, was isolated. Transient transfection studies with different Cyp4b1 promoter constructs indicated that this flanking region exhibits promoter activity when expressed in different cell lines and that a 98-truncated fragment represents the minimal sequence required for basal transcription. Androgen responsiveness was examined by cotransfection with an androgen receptor expression vector, in the presence of androgens. PMID- 12849994 TI - MHC allele-specific binding of a malaria peptide makes it become promiscuous on fitting a glycine residue into pocket 6. AB - Peptide 1585 (EVLYLKPLAGVYRSLKKQLE) has a highly conserved amino-acid sequence located in the Plasmodium falciparum main merozoite surface protein (MSP-1) C terminal region, required for merozoite entry into human erythrocytes and therefore represents a vaccine candidate for P. falciparum malaria. Original sequence-specific binding to five HLA DRB1* alleles (0101, 0102, 0401, 0701, and 1101) revealed this peptide's specific HLA DRB1*0102 allele binding. This peptide's allele-specific binding to HLA DRB1*0102 took on broader specificity for the DRB1*0101, -0401, and -1101 alleles when lysine was replaced by glycine in position 17 (peptide 5198: EVLYLKPLAGVYRSLKG(17)QLE). Binding of the identified G(10)VYRSLKGQLE(20) C-terminal register to these alleles suggests that peptide promiscuous binding relied on fitting Y(12), L(15), and G(17) into P-1, P 4, and P-6, respectively. The implications of the findings and the future of this synthetic vaccine candidate are discussed. PMID- 12849995 TI - Antioxidant inhibits tamoxifen-DNA adducts in endometrial explant culture. AB - Fresh human endometrial explants were incubated for 24h at 37 degrees C with either tamoxifen (10-100 micro M) or the vehicle (0.1% ethanol). Three metabolites namely, alpha-hydroxytamoxifen, 4-hydroxytamoxifen, and N desmethyltamoxifen were identified in the culture media. Tissue size was limited but DNA adducts formed by the alpha-hydroxytamoxifen pathway were detected using authentic alpha-(deoxyguanosyl-N(2)) tamoxifen standards. Relative DNA-adduct levels of 2.45, 1.12, and 0.44 per 10(6) nucleotides were detected following incubations with 100, 25, and 10 micro M tamoxifen, respectively. The concurrent exposure of the explants to 100 micro M tamoxifen with 1mM ascorbic acid reduced the level of alpha-hydroxytamoxifen substantially (68.9%). The formation of tamoxifen-DNA adducts detectable in the explants from the same specimens exposed to 100 micro M tamoxifen with 1mM ascorbic acid were also inhibited. These results support the role of oxidative biotransformation of tamoxifen in the subsequent formation of DNA adducts in this tissue. PMID- 12849996 TI - Alternative splicing and differential expression of P450c17 (CYP17) in gonads during sex transformation in the rice field eel. AB - Several mechanisms were used in determination of the development of the male or female of vertebrates. The genes for determination of sequential hermaphrodite sex are unknown. Here, we reported cloning, alternative splicing, and expression patterns of the CYP17 gene of the rice field eel, a teleost fish with a characteristic of nature sex reversal. The CYP17 gene of the rice field eel was clustered into the CYP17 gene group of all the other vertebrates, especially into the fish subgroup. Four isoforms of the CYP17 were generated in gonads by alternative splicing and polyadenylation. Alternative splicing events of all these isoforms occurred in 3(') regions, which encoded three different sizes (517, 512, and 159aa) of proteins. RT-PCR results indicate specific expression in gonads of these isoforms. Northern blot analysis shows that expression patterns of the CYP17 (dominantly expressed in testis, less in ovary, and the least in ovotestis) are consistent with the sex reversal process of the rice field eel. In situ hybridization further shows its specific expression in germinal lamellae, the gonadal epithelium of the gonads. These findings indicate that CYP17 is differentially regulated in a sex- and developmentally specific manner, suggesting that the CYP17 potentially has important roles in gonad differentiation during sex reversal of the rice field eel. PMID- 12849997 TI - Specificity for inhibitors of metal-substituted methionine aminopeptidase. AB - Methionine aminopeptidases (MetAPs) have been studied in vitro as Co(II) enzymes, but their in vivo metal remains to be defined. While activation of Escherichia coli MetAP (EcMetAP1) by Co(II), Mn(II), and Zn(II) was detectable by a colorimetric Met-S-Gly-Phe assay, significant activation by Ni(II) was shown in a fluorescence Met-AMC assay, in addition to Co(II) and Mn(II) activation. When tested on the metal-substituted EcMetAP1s, a few inhibitors that we obtained recently from a random screening on Co-EcMetAP1 either became much weak or lost activity on Mn- or Zn-EcMetAP1, although they kept inhibitory activity on Ni EcMetAP1. A couple of peptidic inhibitors and the methionine mimetic (3R)-amino (2S)-hydroxyheptanoic acid (AHHpA, 6) maintained moderate activities on Co-, Mn-, Zn-, and Ni-EcMetAP1s. Our results clearly demonstrate that the metal substitution has changed the enzyme specificity for substrates and inhibitors. Therapeutic applications call for inhibitors specific for MetAP with a physiologically relevant metal at its active site. PMID- 12849998 TI - Adenosine A 2B receptors modulate cAMP levels and induce CREB but not ERK1/2 and p38 phosphorylation in rat skeletal muscle cells. AB - The present study examined the existence of the adenosine A(1),A(2A), and A(2B) receptors and the effect of receptor activation on cAMP accumulation and protein phosphorylation in primary rat skeletal muscle cells. Presence of mRNA and protein for all three receptors was demonstrated in both cultured and adult rat skeletal muscle. NECA (10(-9)-10(-4)M) increased the cAMP concentration in cultured muscle cells with an EC(50) of (95% confidence interval)=15 (5.9-25.1) micro M, whereas CGS 21680 (10(-9)-10(-4)M) had no effect on cAMP accumulation. Concentrations of [R]-PIA below 10(-6)M had no effect on cAMP accumulation induced by either isoproterenol or forskolin. NECA resulted in phosphorylation of CREB with an EC(50) of (95% confidence interval)=1.7 (0.40-7.02) micro M, whereas ERK1/2 and p38 phosphorylation was unchanged. The results show that, although the A(1),A(2A), and A(2B) receptors are all present in skeletal muscle cells, the effect of adenosine on adenylyl cyclase activation and phosphorylation of CREB is mainly mediated via the adenosine A(2B) receptor. PMID- 12849999 TI - Identification of epidermal growth factor-responsive genes in normal rat ovarian surface epithelial cells. AB - Alteration in epidermal growth factor receptor (EGFR) family signaling is among the most frequently implicated effectors of human oncogenesis. Overexpression of members of this family of receptors has often been detected in many epithelial tumors and is believed to be associated with an overall poor prognosis in patients with cancer. Therefore, we hypothesized that identification of potential EGF target genes in normal cells will provide a basis for unbiased genetic analysis of this signaling pathway in cancer. We utilized Atlas Rat 1.2 nylon cDNA arrays (Clontech) to determine gene expression changes in normal rat ovarian surface epithelial (ROSE) cells following EGF treatment. The results indicate activation of genes involved in a wide variety of cellular mechanisms, including regulation of cell cycle and proliferation, apoptosis, and protein turnover. In addition, using an in vitro model of ovarian cancer, we demonstrated that malignant transformation of ROSE cells resulted in alteration of downstream effectors of the EGFR pathway, as exemplified by aberrant expression of p66Shc, c Jun, c-Myc, c-Fos, Lot1, p21Cip/Waf, and cdc25A. These data suggest that knowledge of the downstream genetic lesions, which may result in loss of growth factor requirement of the affected cells, will be crucial for the selection of the EGFR pathway as an effective target for cancer therapy. PMID- 12850000 TI - A peptide mimetic of an anti-CD4 monoclonal antibody by rational design. AB - The development of rational methods to design 'continuous' sequence mimetics of discontinuous regions of protein sequence has, to now, been only marginally successful. This has been largely due to the difficulty of constraining the recognition elements of a mimetic structure to the relative conformational and spatial orientations present in the parent molecule. Using peptide mapping to determine 'active' antigen recognition residues, molecular modeling, and a molecular dynamics trajectory analysis, we have developed a peptide mimic of an anti-CD4 antibody, containing antigen contact residues from multiple CDRs. The design described is a 27-residue peptide formed by juxtaposition of residues from 5 CDR regions. It displays an affinity for the antigen (CD4) of 0.9nM, compared to 2nM for the parent antibody ST40. Nevertheless, the mimetic shows low biological activity in an anti-retroviral assay. PMID- 12850002 TI - Advances in Neural Networks Research. Proceedings of the 2003 International Joint Conference on Neural Networks (IJCNN). PMID- 12850001 TI - Accumulation of p27 KIP1 is associated with BMP2-induced growth arrest and neuronal differentiation of human neuroblastoma-derived cell lines. AB - Bone morphogenetic proteins (BMPs) play an essential role in cell fate determination. In this study, we found that BMP2 treatment resulted in growth arrest and differentiation in human neuroblastoma-derived cell lines, SH-SY5Y and RTBM1. Within 30min of BMP2 exposure, phosphorylation of Smad1/5 was observed in these cell lines. In RTBM1 cells, BMP2-induced differentiation was accompanied by a significant decrease in the expression level of DAN, an antagonist of BMP in frog embryos. Immunoblot analysis revealed that BMP2 treatment caused a down regulation of p53 family members and hence of cyclin-dependent kinase inhibitor p21(WAF1). We found a significant accumulation of p27(KIP1) in response to BMP2, whereas the expression level of Skp2, which is required for ubiquitin-dependent p27(KIP1) degradation, was decreased during this differentiation process. Our results suggest that p27(KIP1) contributes to the BMP-induced growth arrest and neuronal differentiation of neuroblastoma, and BMP treatment might provide a new therapeutic strategy. PMID- 12850003 TI - Adaptive force generation for precision-grip lifting by a spectral timing model of the cerebellum. AB - We modeled adaptive generation of precision grip forces during object lifting. The model presented adjusts reactive and anticipatory grip forces to a level just above that needed to stabilize lifted objects in the hand. The model obeys principles of cerebellar structure and function by using slip sensations as error signals to adapt phasic motor commands to tonic force generators associated with output synergies controlling grip aperture. The learned phasic commands are weight- and texture-dependent. Simulations of the new circuit model reproduce key aspects of experimental observations of force application. Over learning trials, the onset of grip force buildup comes to lead the load force buildup, and the rate-of-rise of grip force, but not load force, scales inversely with the friction of the object. PMID- 12850004 TI - Radial basis function neural networks for nonlinear Fisher discrimination and Neyman-Pearson classification. AB - We propose a novel technique for the design of radial basis function (RBF) neural networks (NNs). To select various RBF parameters, the class membership information of training samples is utilized to produce new cluster classes. This allows emphasis of classification performance for certain class data rather than best overall classification. This allows us to control performance as desired and to approximate Neyman-Pearson classification. We also show that by properly choosing the desired output neuron levels, then the RBF hidden to output layer performs Fisher discrimination analysis, and that the full system performs a nonlinear Fisher analysis. Data on an agricultural product inspection problem and on synthetic data confirm the effectiveness of these methods. PMID- 12850005 TI - Intrinsic generalization analysis of low dimensional representations. AB - Low dimensional representations of images impose equivalence relations in the image space; the induced equivalence class of an image is named as its intrinsic generalization. The intrinsic generalization of a representation provides a novel way to measure its generalization and leads to more fundamental insights than the commonly used recognition performance, which is heavily influenced by the choice of training and test data. We demonstrate the limitations of linear subspace representations by sampling their intrinsic generalization, and propose a nonlinear representation that overcomes these limitations. The proposed representation projects images nonlinearly into the marginal densities of their filter responses, followed by linear projections of the marginals. We use experiments on large datasets to show that the representations that have better intrinsic generalization also lead to better recognition performance. PMID- 12850007 TI - Subject independent facial expression recognition with robust face detection using a convolutional neural network. AB - Reliable detection of ordinary facial expressions (e.g. smile) despite the variability among individuals as well as face appearance is an important step toward the realization of perceptual user interface with autonomous perception of persons. We describe a rule-based algorithm for robust facial expression recognition combined with robust face detection using a convolutional neural network. In this study, we address the problem of subject independence as well as translation, rotation, and scale invariance in the recognition of facial expression. The result shows reliable detection of smiles with recognition rate of 97.6% for 5600 still images of more than 10 subjects. The proposed algorithm demonstrated the ability to discriminate smiling from talking based on the saliency score obtained from voting visual cues. To the best of our knowledge, it is the first facial expression recognition model with the property of subject independence combined with robustness to variability in facial appearance. PMID- 12850006 TI - Application of four-layer neural network on information extraction. AB - This paper applies neural network to extract marsh information. An adaptive back propagation algorithm based on a robust error function is introduced to build a four-layer neural network, and it is used to classify Thematic Mapper (TM) image of Zhalong Wetland in China and then extract marsh information. Comparing marsh information extraction results of the four-layer neural network with three-layer neural network and the maximum likelihood classifier, conclusion can be drawn as follows: the structure of the four-layer neural network and the adaptive back propagation algorithm based on the robust error function is effective to extract marsh information. The four-layer neural network adopted in this paper succeeded in building the complex model of TM image, and it avoided the problem of great storage of remotely sensed data, and the adaptive back-propagation algorithm speeded up the descending of error. Above all, the four-layer neural network is superior to the three-layer neural network and the maximum likelihood classifier in the accuracy of the total classification and marsh information extraction. PMID- 12850008 TI - A generalized feedforward neural network architecture for classification and regression. AB - This article presents a new generalized feedforward neural network (GFNN) architecture for pattern classification and regression. The GFNN architecture uses as the basic computing unit a generalized shunting neuron (GSN) model, which includes as special cases the perceptron and the shunting inhibitory neuron. GSNs are capable of forming complex, nonlinear decision boundaries. This allows the GFNN architecture to easily learn some complex pattern classification problems. In this article the GFNNs are applied to several benchmark classification problems, and their performance is compared to the performances of SIANNs and multilayer perceptrons. Experimental results show that a single GSN can outperform both the SIANN and MLP networks. PMID- 12850009 TI - Hierarchical cognitive maps. AB - We describe a computational model of spatial navigation based on experimental studies conducted with human participants. The model builds and uses a hierarchical cognitive map of a large environment. Computer simulations show that the model correctly describes experimental results including hierarchical organization of space and distance estimation. Furthermore, the model predicts that reaction time for distance estimation varies nonlinearly with distance. PMID- 12850010 TI - Modeling goal-directed spatial navigation in the rat based on physiological data from the hippocampal formation. AB - We investigated the importance of hippocampal theta oscillations and the significance of phase differences of theta modulation in the cortical regions that are involved in goal-directed spatial navigation. Our models used representations of entorhinal cortex layer III (ECIII), hippocampus and prefrontal cortex (PFC) to guide movements of a virtual rat in a virtual environment. The model encoded representations of the environment through long term potentiation of excitatory recurrent connections between sequentially spiking place cells in ECIII and CA3. This encoding required buffering of place cell activity, which was achieved by a short-term memory (STM) in EC that was regulated by theta modulation and allowed synchronized reactivation with encoding phases in ECIII and CA3. Inhibition at a specific theta phase deactivated the oldest item in the buffer when new input was presented to a full STM buffer. A 180 degrees phase difference separated retrieval and encoding in ECIII and CA3, which enabled us to simulate data on theta phase precession of place cells. Retrieval of known paths was elicited in ECIII by input at the retrieval phase from PFC working memory for goal location, requiring strict theta phase relationships with PFC. Known locations adjacent to the virtual rat were retrieved in CA3. Together, input from ECIII and CA3 activated predictive spiking in cells in CA1 for the next desired place on a shortest path to a goal. Consistent with data, place cell activity in CA1 and CA3 showed smaller place fields than in ECIII. PMID- 12850011 TI - An efficient training algorithm for dynamic synapse neural networks using trust region methods. AB - We formulate the dynamic synapse neural network from the averaged activity of local population of neurons perspective. We have applied the trust region nonlinear optimization approach to train the network and show the new learning method effectiveness in comparison to the genetic algorithms by optimizing large scale networks. PMID- 12850012 TI - Temporal binding as an inducer for connectionist recruitment learning over delayed lines. AB - The temporal correlation hypothesis proposes using distributed synchrony for the binding of different stimulus features. However, synchronized spikes must travel over cortical circuits that have varying length pathways, leading to mismatched arrival times. This raises the question of how initial stimulus-dependent synchrony might be preserved at a destination binding site. Earlier, we proposed constraints on tolerance and segregation parameters for a phase-coding approach, within cortical circuits, to address this question [Proceedings of the International Joint Conference on Neural Networks, Washington, DC, 2001]. The purpose of the present paper is twofold. First, we conduct simulation experiments to test the proposed constraints. Second, we explore the practicality of temporal binding to drive a process of long-term memory formation based on a recruitment learning method [Biol. Cybernet. 46 (1982) 27]. A network based on Valiant's neuroidal architecture [Circuits of the mind, 1994] is used to demonstrate the coalition between temporal binding and recruitment. Complementing similar approaches, we implement a continuous-time learning procedure allowing computation with spiking neurons. The viability of the proposed binding scheme is investigated by conducting simulation studies which examine binding errors. In the simulation, binding errors cause the perception of illusory conjunctions among features belonging to separate objects. Our results indicate that when tolerance and segregation parameters obey our proposed constraints, the assemblies of correct bindings are dominant over assemblies of spurious bindings in reasonable operating conditions. PMID- 12850013 TI - Developments in understanding neuronal spike trains and functional specializations in brain regions. AB - Understanding information processing at the neuronal level would provide valuable insights to computational intelligence research and computational neuroscience. In particular, understanding constraints on neuronal spike trains would provide indication about the type of syntactic rules used by neurons when processing information. A recent discovery, reported here, was made through analyzing microelectrode recordings (MER) made during surgical procedure in humans. Analysis of MERs of extracellular neuronal activity has gained increasing interest due to potential improvements to surgical techniques involving ablation or placement of deep brain stimulators, done in the treatment of advanced Parkinson's disease. Important to these procedures is the identification of different brain structures such as the globus pallidus internus from the spike train being recorded from the intracranial probe tip during surgery. Spike train data gathered during surgical procedure from multiple patients were processed using a novel feature extraction method reported here. Distinct structures within the spike trains were identified and used to build an effective brain region classifier. The extracted features upon analysis provide some insight into the 'syntactic' constraint on spike trains. PMID- 12850014 TI - Shaping up simple cell's receptive field of animal vision by ICA and its application in navigation system. AB - Scientists and experts have explored the mechanism of visual systems for decades for smart image processing and pattern recognition in order to satisfy sophisticated engineering applications. In this paper we apply independent component analyses' (ICA) unsupervised learning to natural images, topography images and other special environment images for demonstrating the simple-cell's process in animal vision. Our results confirm an early biological experiment (Nature 228 (1970) 419) about the growth of simple cells in cat's V1 area. Furthermore, by applying ICA methodology and the simplex algorithm, the unsupervised neural synapse's learning can obtain the receptive fields in visual cortex and can simulate the growth of the visual cortex of young animal in the special environment. These findings imply that an input image can be efficiently represented by ICA bases. An application of image matching in the navigation by ICA is shown that the animal visual system method is indeed better than those classical methods at least more than 5%. PMID- 12850015 TI - eLoom and Flatland: specification, simulation and visualization engines for the study of arbitrary hierarchical neural architectures. AB - eLoom is an open source graph simulation software tool, developed at the University of New Mexico (UNM), that enables users to specify and simulate neural network models. Its specification language and libraries enables users to construct and simulate arbitrary, potentially hierarchical network structures on serial and parallel processing systems. In addition, eLoom is integrated with UNM's Flatland, an open source virtual environments development tool to provide real-time visualizations of the network structure and activity. Visualization is a useful method for understanding both learning and computation in artificial neural networks. Through 3D animated pictorially representations of the state and flow of information in the network, a better understanding of network functionality is achieved. ART-1, LAPART-II, MLP, and SOM neural networks are presented to illustrate eLoom and Flatland's capabilities. PMID- 12850016 TI - Associative morphological memories based on variations of the kernel and dual kernel methods. AB - Morphological associative memories (MAMs) belong to the class of morphological neural networks. The recording scheme used in the original MAM models is similar to the correlation recording recipe. Recording is achieved by means of a maximum (MXY model) or minimum (WXY model) of outer products. Notable features of autoassociative morphological memories (AMMs) include optimal absolute storage capacity and one-step convergence. Heteroassociative morphological memories (HMMs) do not have these properties and are not very well understood. The fixed points of AMMs can be characterized exactly in terms of the original patterns. Unfortunately, AMM fixed points include a large number of spurious memories. In this paper, we combine the MXX model and variations of the kernel method to produce new autoassociative and heteroassociative memories. We also introduce a dual kernel method. A new, dual model is given by a combination of the WXX model and a variation of the dual kernel method. The new MAM models exhibit better error correction capabilities than MXX and WXX and a reduced number of spurious memories which can be easily described in terms of the fundamental memories. PMID- 12850017 TI - Adaptive double self-organizing maps for clustering gene expression profiles. AB - This paper introduces a new model of self-organizing map (SOM) known as adaptive double self-organizing map (ADSOM). ADSOM has a flexible topology and performs data partitioning and cluster visualization simultaneously without requiring a priori knowledge about the number of clusters. It combines features of the popular SOM with two-dimensional position vectors, which serve as a visualization tool to detect the number of clusters present in the data. ADSOM updates its free parameters and allows convergence of its position vectors to a fairly consistent number of clusters provided its initial number of nodes is greater than the expected number of clusters. A novel index is introduced based on hierarchical clustering of the final locations of position vectors. The index allows automated detection of the number of clusters, thereby reducing human error that could be incurred from counting clusters visually. To test ADSOM's consistency in data partitioning, we examine the number of common profiles found in the clusters that were obtained by varying the initial number of nodes. This provides a confidence measure for the clusters formed by ADSOM and illustrates the effect of different initial number of nodes on data partitioning. The reliance of ADSOM in identifying number of clusters is demonstrated by applying it to publicly available yeast gene expression data. PMID- 12850018 TI - An accelerated procedure for recursive feature ranking on microarray data. AB - We describe a new wrapper algorithm for fast feature ranking in classification problems. The Entropy-based Recursive Feature Elimination (E-RFE) method eliminates chunks of uninteresting features according to the entropy of the weights distribution of a SVM classifier. With specific regard to DNA microarray datasets, the method is designed to support computationally intensive model selection in classification problems in which the number of features is much larger than the number of samples. We test E-RFE on synthetic and real data sets, comparing it with other SVM-based methods. The speed-up obtained with E-RFE supports predictive modeling on high dimensional microarray data. PMID- 12850019 TI - Pattern completion through phase coding in population neurodynamics. AB - This article presents an alternative phase coding mechanism for Freeman's KIII model of population neurodynamics. Motivated by experimental evidence that supports the existence of a neural code based on synchronous oscillations, we propose an analogy between synchronization in neural populations and phase locking in KIII channels. An efficient method is proposed to extract phase differences across granule channels from their state-space trajectories. First, the scale invariance of the KIII model with respect to phase information is established. The phase code is then compared against the conventional amplitude code in terms of their bit-wise and across-fiber pattern recovery capabilities using decision-theoretic principles and a Hamming-distance classifier. Graph isomorphism in the Hebbian connections is exploited to perform an exhaustive evaluation of patterns on an 8-channel KIII model. Simulation results show that phase information outperforms amplitude information in the recovery of incomplete or corrupted stimuli. PMID- 12850020 TI - Passive dendritic integration heavily affects spiking dynamics of recurrent networks. AB - According to dendritic cable theory, proximal synapses give rise to inputs with short delay, high amplitude, and short duration. In contrast, inputs from distal synapses have long delays, low amplitude, and long duration. Nevertheless, large scale neural networks are seldom built with realistically layered synaptic architectures and corresponding electrotonic parameters. Here, we use a simple model to investigate the spike response dynamics of networks with different electrotonic structures. The networks consist of a layer of neurons receiving a sparse feedforward projection from a set of inputs, as well as sparse recurrent connections from within the layer. Firing patterns are set in the inputs, and recorded from the neuron (output) layer. The feedforward and recurrent synapses are independently set as proximal or distal, representing dendritic connections near or far from the soma, respectively. Analyses of firing dynamics indicate that recurrent distal synapses tend to concentrate network activity in fewer neurons, while proximal recurrent synapses result in a more homogeneous activity distribution. In addition, when the feedforward input is regular (spiking or bursting) and asynchronous, the output is regular if recurrent synapses are more distal than feedforward ones, and irregular in the opposite configuration. Finally, the amplitude of network fluctuations in response to asynchronous input is lower if feedforward and recurrent synapses are electrotonically distant from one another (in either configuration). In conclusion, electrotonic effects reflecting different dendritic positions of synaptic inputs significantly influence network dynamics. PMID- 12850021 TI - Abductive reasoning with recurrent neural networks. AB - Abduction is the process of proceeding from data describing a set of observations or events, to a set of hypotheses which best explains or accounts for the data. Cost-based abduction (CBA) is a formalism in which evidence to be explained is treated as a goal to be proven, proofs have costs based on how much needs to be assumed to complete the proof, and the set of assumptions needed to complete the least-cost proof are taken as the best explanation for the given evidence. In previous work, we presented a method for using high order recurrent networks to find least cost proofs for CBA instances. Here, we present a method that significantly reduces the size of the neural network that is produced for a given CBA instance. We present experimental results describing the performance of this method and comparing its performance to that of the previous method. PMID- 12850022 TI - Neural networks with chaotic recursive nodes: techniques for the design of associative memories, contrast with Hopfield architectures, and extensions for time-dependent inputs. AB - This paper addresses the coding and storage of information in neural architectures with bifurcating recursive nodes that exhibit chaotic dynamics. It describes architectures of coupled recursive processing elements (RPEs) used to store binary strings, discusses the choices of network parameters related to the coding of zeros and ones, and analyzes several aspects of the network operation in implementing associative memories through populations of logistic maps. Experiments for the performance evaluation of these memories are described, and results addressing the operation under digital noise (flipped bits) and analog noise added to the prompting pattern are presented and analyzed. Quantitative aspects related to the representation of binary strings through cyclic states are equated, and then related to the planning and analysis of several experiments. A simple pre-processing procedure useful in situations of prompting conditions with analog noise is proposed, and the resultant increase in recovery performance presented. The performance of the RPEs associative networks is contrasted with the performance of Hopfield associative memories, and the situations where the RPEs networks present significant superiority are identified. An extended version of the proposed architecture, which allows to address the issues of time dependent inputs and analog inputs, is analyzed in detail. Experimental results are presented, and the role of this extended architecture in providing mechanisms for modular RPEs architectures is pointed out. PMID- 12850023 TI - Simple and conditioned adaptive behavior from Kalman filter trained recurrent networks. AB - We illustrate the ability of a fixed-weight neural network, trained with Kalman filter methods, to perform tasks that are usually entrusted to an explicitly adaptive system. Following a simple example, we demonstrate that such a network can be trained to exhibit input-output behavior that depends on which of two conditioning tasks was performed a substantial number of time steps in the past. This behavior can also be made to survive an intervening interference task. PMID- 12850024 TI - Learning robot actions based on self-organising language memory. AB - In the MirrorBot project we examine perceptual processes using models of cortical assemblies and mirror neurons to explore the emergence of semantic representations of actions, percepts and concepts in a neural robot. The hypothesis under investigation is whether a neural model will produce a life-like perception system for actions. In this context we focus in this paper on how instructions for actions can be modeled in a self-organising memory. Current approaches for robot control often do not use language and ignore neural learning. However, our approach uses language instruction and draws from the concepts of regional distributed modularity, self-organisation and neural assemblies. We describe a self-organising model that clusters actions into different locations depending on the body part they are associated with. In particular, we use actual sensor readings from the MIRA robot to represent semantic features of the action verbs. Furthermore, we outline a hierarchical computational model for a self-organising robot action control system using language for instruction. PMID- 12850025 TI - Autonomous mental development in high dimensional context and action spaces. AB - Autonomous Mental Development (AMD) of robots opened a new paradigm for developing machine intelligence, using neural network type of techniques and it fundamentally changed the way an intelligent machine is developed from manual to autonomous. The work presented here is a part of SAIL (Self-Organizing Autonomous Incremental Learner) project which deals with autonomous development of humanoid robot with vision, audition, manipulation and locomotion. The major issue addressed here is the challenge of high dimensional action space (5-10) in addition to the high dimensional context space (hundreds to thousands and beyond), typically required by an AMD machine. This is the first work that studies a high dimensional (numeric) action space in conjunction with a high dimensional perception (context state) space, under the AMD mode. Two new learning algorithms, Direct Update on Direction Cosines (DUDC) and High Dimensional Conjugate Gradient Search (HCGS), are developed, implemented and tested. The convergence properties of both the algorithms and their targeted applications are discussed. Autonomous learning of speech production under reinforcement learning is studied as an example. PMID- 12850026 TI - Chaos control and synchronization, with input saturation, via recurrent neural networks. AB - This paper deals with the adaptive tracking problem of non-linear systems in presence of unknown parameters, unmodelled dynamics and input saturation. A high order recurrent neural network is used in order to identify the unknown system and a learning law is obtained using the Lyapunov methodology. Then a stabilizing control law for the reference tracking error dynamics is developed using the Lyapunov methodology and the Sontag control law. Tracking error boundedness is established as a function of a design parameter. The new approach is illustrated by examples of complex dynamical systems: chaos control and synchronization. PMID- 12850027 TI - Proper orthogonal decomposition based optimal neurocontrol synthesis of a chemical reactor process using approximate dynamic programming. AB - The concept of approximate dynamic programming and adaptive critic neural network based optimal controller is extended in this study to include systems governed by partial differential equations. An optimal controller is synthesized for a dispersion type tubular chemical reactor, which is governed by two coupled nonlinear partial differential equations. It consists of three steps: First, empirical basis functions are designed using the 'Proper Orthogonal Decomposition' technique and a low-order lumped parameter system to represent the infinite-dimensional system is obtained by carrying out a Galerkin projection. Second, approximate dynamic programming technique is applied in a discrete time framework, followed by the use of a dual neural network structure called adaptive critics, to obtain optimal neurocontrollers for this system. In this structure, one set of neural networks captures the relationship between the state variables and the control, whereas the other set captures the relationship between the state and the costate variables. Third, the lumped parameter control is then mapped back to the spatial dimension using the same basis functions to result in a feedback control. Numerical results are presented that illustrate the potential of this approach. It should be noted that the procedure presented in this study can be used in synthesizing optimal controllers for a fairly general class of nonlinear distributed parameter systems. PMID- 12850028 TI - Numerical solution of elliptic partial differential equation using radial basis function neural networks. AB - In this paper a neural network for solving partial differential equations is described. The activation functions of the hidden nodes are the radial basis functions (RBF) whose parameters are learnt by a two-stage gradient descent strategy. A new growing RBF-node insertion strategy with different RBF is used in order to improve the net performances. The learning strategy is able to save computational time and memory space because of the selective growing of nodes whose activation functions consist of different RBFs. An analysis of the learning capabilities and a comparison of the net performances with other approaches have been performed. It is shown that the resulting network improves the approximation results. PMID- 12850029 TI - Statistical efficiency of adaptive algorithms. AB - The statistical efficiency of a learning algorithm applied to the adaptation of a given set of variable weights is defined as the ratio of the quality of the converged solution to the amount of data used in training the weights. Statistical efficiency is computed by averaging over an ensemble of learning experiences. A high quality solution is very close to optimal, while a low quality solution corresponds to noisy weights and less than optimal performance. In this work, two gradient descent adaptive algorithms are compared, the LMS algorithm and the LMS/Newton algorithm. LMS is simple and practical, and is used in many applications worldwide. LMS/Newton is based on Newton's method and the LMS algorithm. LMS/Newton is optimal in the least squares sense. It maximizes the quality of its adaptive solution while minimizing the use of training data. Many least squares adaptive algorithms have been devised over the years, but no other least squares algorithm can give better performance, on average, than LMS/Newton. LMS is easily implemented, but LMS/Newton, although of great mathematical interest, cannot be implemented in most practical applications. Because of its optimality, LMS/Newton serves as a benchmark for all least squares adaptive algorithms. The performances of LMS and LMS/Newton are compared, and it is found that under many circumstances, both algorithms provide equal performance. For example, when both algorithms are tested with statistically nonstationary input signals, their average performances are equal. When adapting with stationary input signals and with random initial conditions, their respective learning times are on average equal. However, under worst-case initial conditions, the learning time of LMS can be much greater than that of LMS/Newton, and this is the principal disadvantage of the LMS algorithm. But the strong points of LMS are ease of implementation and optimal performance under important practical conditions. For these reasons, the LMS algorithm has enjoyed very widespread application. It is used in almost every modem for channel equalization and echo cancelling. Furthermore, it is related to the famous backpropagation algorithm used for training neural networks. PMID- 12850030 TI - On structure-exploiting trust-region regularized nonlinear least squares algorithms for neural-network learning. AB - This paper briefly introduces our numerical linear algebra approaches for solving structured nonlinear least squares problems arising from 'multiple-output' neural network (NN) models. Our algorithms feature trust-region regularization, and exploit sparsity of either the 'block-angular' residual Jacobian matrix or the 'block-arrow' Gauss-Newton Hessian (or Fisher information matrix in statistical sense) depending on problem scale so as to render a large class of NN-learning algorithms 'efficient' in both memory and operation costs. Using a relatively large real-world nonlinear regression application, we shall explain algorithmic strengths and weaknesses, analyzing simulation results obtained by both direct and iterative trust-region algorithms with two distinct NN models: 'multilayer perceptrons' (MLP) and 'complementary mixtures of MLP-experts' (or neuro-fuzzy modular networks). PMID- 12850031 TI - Stochastic resonance in noisy threshold neurons. AB - Stochastic resonance occurs when noise improves how a nonlinear system performs. This paper presents two general stochastic-resonance theorems for threshold neurons that process noisy Bernoulli input sequences. The performance measure is Shannon mutual information. The theorems show that small amounts of independent additive noise can increase the mutual information of threshold neurons if the neurons detect subthreshold signals. The first theorem shows that this stochastic resonance effect holds for all finite-variance noise probability density functions that obey a simple mean constraint that the user can control. A corollary shows that this stochastic-resonance effect occurs for the important family of (right-sided) gamma noise. The second theorem shows that this effect holds for all infinite-variance noise types in the broad family of stable distributions. Stable bell curves can model extremely impulsive noise environments. So the second theorem shows that this stochastic-resonance effect is robust against violent fluctuations in the additive noise process. PMID- 12850032 TI - Quantum optimization for training support vector machines. AB - Refined concepts, such as Rademacher estimates of model complexity and nonlinear criteria for weighting empirical classification errors, represent recent and promising approaches to characterize the generalization ability of Support Vector Machines (SVMs). The advantages of those techniques lie in both improving the SVM representation ability and yielding tighter generalization bounds. On the other hand, they often make Quadratic-Programming algorithms no longer applicable, and SVM training cannot benefit from efficient, specialized optimization techniques. The paper considers the application of Quantum Computing to solve the problem of effective SVM training, especially in the case of digital implementations. The presented research compares the behavioral aspects of conventional and enhanced SVMs; experiments in both a synthetic and real-world problems support the theoretical analysis. At the same time, the related differences between Quadratic Programming and Quantum-based optimization techniques are considered. PMID- 12850033 TI - On the quality of ART1 text clustering. AB - There is a large and continually growing quantity of electronic text available, which contain essential human and organization knowledge. An important research endeavor is to study and develop better ways to access this knowledge. Text clustering is a popular approach to automatically organize textual document collections by topics to help users find the information they need. Adaptive Resonance Theory (ART) neural networks possess several interesting properties that make them appealing in the area of text clustering. Although ART has been used in several research works as a text clustering tool, the level of quality of the resulting document clusters has not been clearly established yet. In this paper, we present experimental results with binary ART that address this issue by determining how close clustering quality is to an upper bound on clustering quality. PMID- 12850034 TI - Extension neural network and its applications. AB - In this paper, a novel extension neural network (ENN) is proposed. This new neural network is a combination of extension theory and neural network. It uses an extension distance (ED) to measure the similarity between data and cluster center. The learning speed of the proposed ENN is shown to be faster than the traditional neural networks and other fuzzy classification methods. Moreover, the new scheme has been proved to have high accuracy and less memory consumption. Experimental results from two different examples verify the effectiveness and applicability of the proposed work. PMID- 12850035 TI - Fuzzy least squares support vector machines for multiclass problems. AB - In least squares support vector machines (LS-SVMs), the optimal separating hyperplane is obtained by solving a set of linear equations instead of solving a quadratic programming problem. But since SVMs and LS-SVMs are formulated for two class problems, unclassifiable regions exist when they are extended to multiclass problems. In this paper, we discuss fuzzy LS-SVMs that resolve unclassifiable regions for multiclass problems. We define a membership function in the direction perpendicular to the optimal separating hyperplane that separates a pair of classes. Using the minimum or average operation for these membership functions, we define a membership function for each class. Using some benchmark data sets, we show that recognition performance of fuzzy LS-SVMs with the minimum operator is comparable to that of fuzzy SVMs, but fuzzy LS-SVMs with the average operator showed inferior performance. PMID- 12850036 TI - Evolving efficient learning algorithms for binary mappings. AB - Gradient descent training of sigmoidal feed-forward neural networks on binary mappings often gets stuck with some outputs totally wrong. This is because a sum squared-error cost function leads to weight updates that depend on the derivative of the output sigmoid which goes to zero as the output approaches maximal error. Although it is easy to understand the cause, the best remedy is not so obvious. Common solutions involve modifying the training data, deviating from true gradient descent, or changing the cost function. In general, finding the best learning procedures for particular classes of problem is difficult because each usually depends on a number of interacting parameters that need to be set to optimal values for a fair comparison. In this paper I shall use simulated evolution to optimise all the relevant parameters, and come to a clear conclusion concerning the most efficient approach for learning binary mappings. PMID- 12850037 TI - A network for recursive extraction of canonical coordinates. AB - A network structure for canonical coordinate decomposition is presented. The network consists of two single-layer linear subnetworks that together extract the canonical coordinates of two data channels. The connection weights of the networks are trained by a stochastic gradient descent learning algorithm. Each subnetwork features a hierarchical set of lateral connections among its outputs. The lateral connections perform a deflation process that subtracts the contributions of the already extracted coordinates from the input data subspace. This structure allows for adding new nodes for extracting additional canonical coordinates without the need for retraining the previous nodes. The performance of the network is evaluated on a synthesized data set. PMID- 12850038 TI - Automatic basis selection techniques for RBF networks. AB - This paper proposes a generic criterion that defines the optimum number of basis functions for radial basis function (RBF) neural networks. The generalization performance of an RBF network relates to its prediction capability on independent test data. This performance gives a measure of the quality of the chosen model. An RBF network with an overly restricted basis gives poor predictions on new data, since the model has too little flexibility (yielding high bias and low variance). By contrast, an RBF network with too many basis functions also gives poor generalization performance since it is too flexible and fits too much of the noise on the training data (yielding low bias but high variance). Bias and variance are complementary quantities, and it is necessary to assign the number of basis function optimally in order to achieve the best compromise between them. In this paper we use Stein's unbiased risk estimator to derive an analytical criterion for assigning the appropriate number of basis functions. Two cases of known and unknown noise have been considered and the efficacy of this criterion in both situations is illustrated experimentally. The paper also shows an empirical comparison between this method and two well known classical methods, cross validation and the Bayesian information criterion, BIC. PMID- 12850039 TI - Data smoothing regularization, multi-sets-learning, and problem solving strategies. AB - First, we briefly introduce the basic idea of data smoothing regularization, which was firstly proposed by Xu [Brain-like computing and intelligent information systems (1997) 241] for parameter learning in a way similar to Tikhonov regularization but with an easy solution to the difficulty of determining an appropriate hyper-parameter. Also, the roles of this regularization are demonstrated on Gaussian-mixture via smoothed versions of the EM algorithm, the BYY model selection criterion, adaptive harmony algorithm as well as its related Rival penalized competitive learning. Second, these studies are extended to a mixture of reconstruction errors of Gaussian types, which provides a new probabilistic formulation for the multi-sets learning approach [Proc. IEEE ICNN94 I (1994) 315] that learns multiple objects in typical geometrical structures such as points, lines, hyperplanes, circles, ellipses, and templates of given shapes. Finally, insights are provided on three problem solving strategies, namely the competition-penalty adaptation based learning, the global evidence accumulation based selection, and the guess-test based decision, with a general problem solving paradigm suggested. PMID- 12850040 TI - Million city traveling salesman problem solution by divide and conquer clustering with adaptive resonance neural networks. AB - The Traveling Salesman Problem (TSP) is a very hard optimization problem in the field of operations research. It has been shown to be NP-complete, and is an often-used benchmark for new optimization techniques. One of the main challenges with this problem is that standard, non-AI heuristic approaches such as the Lin Kernighan algorithm (LK) and the chained LK variant are currently very effective and in wide use for the common fully connected, Euclidean variant that is considered here. This paper presents an algorithm that uses adaptive resonance theory (ART) in combination with a variation of the Lin-Kernighan local optimization algorithm to solve very large instances of the TSP. The primary advantage of this algorithm over traditional LK and chained-LK approaches is the increased scalability and parallelism allowed by the divide-and-conquer clustering paradigm. Tours obtained by the algorithm are lower quality, but scaling is much better and there is a high potential for increasing performance using parallel hardware. PMID- 12850041 TI - A practical sub-space adaptive filter. AB - A Sub-Space Adaptive Filter (SSAF) model is developed using, as a basis, the Modified Probabilistic Neural Network (MPNN) and its extension the Tuneable Approximate Piecewise Linear Regression (TAPLR) model. The TAPLR model can be adjusted by a single smoothing parameter continuously from the best piecewise linear model in each sub-space to the best approximately piecewise linear model over the whole data space. A suitable value in between ensures that all neighbouring piecewise linear models merge together smoothly at their boundaries. This model was developed by altering the form of the MPNN, a network used for general nonlinear regression. The MPNNs special structure allows it to be easily used to model a process by appropriately weighting piecewise linear models associated with each of the network's radial basis functions. The model has now been further extended to allow each piecewise linear model section to be adapted separately as new data flows through it. By doing this, the proposed SSAF model represents a learning/filtering method for nonlinear processes that provides one solution to the stability/plasticity dilemma associated with standard adaptive filters. PMID- 12850042 TI - Pharmacodynamic population analysis in chronic renal failure using artificial neural networks--a comparative study. AB - This work presents a pharmacodynamic population analysis in chronic renal failure patients using Artificial Neural Networks (ANNs). In pursuit of an effective and cost-efficient strategy for drug delivery in patients with renal failure, two different types of ANN are applied to perform drug dose-effect modeling and their performance compared. Applied in a clinical environment, such models will allow for prediction of patient response to the drug at the effect site and, subsequently, for adjusting the dosing regimen. PMID- 12850043 TI - Electronic nose based tea quality standardization. AB - In this paper we have used a metal oxide sensor (MOS) based electronic nose (EN) to analyze five tea samples with different qualities, namely, drier month, drier month again over-fired, well fermented normal fired in oven, well fermented overfired in oven, and under fermented normal fired in oven. The flavour of tea is determined mainly by its taste and smell, which is generated by hundreds of Volatile Organic Compounds (VOCs) and Non-Volatile Organic Compounds present in tea. These VOCs are present in different ratios and determine the quality of the tea. For example Assamica (Sri Lanka and Assam Tea) and Assamica Sinesis (Darjeeling and Japanese Tea) are two different species of tea giving different flavour notes. Tea flavour is traditionally measured through the use of a combination of conventional analytical instrumentation and human or ganoleptic profiling panels. These methods are expensive in terms of time and labour and also inaccurate because of a lack of either sensitivity or quantitative information. In this paper an investigation has been made to determine the flavours of different tea samples using an EN and to explore the possibility of replacing existing analytical and profiling panel methods. The technique uses an array of 4 MOSs, each of, which has an electrical resistance that has partial sensitivity to the headspace of tea. The signals from the sensor array are then conditioned by suitable interface circuitry. The data were processed using Principal Components Analysis (PCA), Fuzzy C Means algorithm (FCM). We also explored the use of a Self-Organizing Map (SOM) method along with a Radial Basis Function network (RBF) and a Probabilistic Neural Network classifier. Using FCM and SOM feature extraction techniques along with RBF neural network we achieved 100% correct classification for the five different tea samples with different qualities. These results prove that our EN is capable of discriminating between the flavours of teas manufactured under different processing conditions, viz. over-fermented, over-fired, under fermented, etc. PMID- 12850044 TI - A novel neural network-based survival analysis model. AB - A feedforward neural network architecture aimed at survival probability estimation is presented which generalizes the standard, usually linear, models described in literature. The network builds an approximation to the survival probability of a system at a given time, conditional on the system features. The resulting model is described in a hierarchical Bayesian framework. Experiments with synthetic and real world data compare the performance of this model with the commonly used standard ones. PMID- 12850045 TI - Divide-and-conquer approach for brain machine interfaces: nonlinear mixture of competitive linear models. AB - This paper proposes a divide-and-conquer strategy for designing brain machine interfaces. A nonlinear combination of competitively trained local linear models (experts) is used to identify the mapping from neuronal activity in cortical areas associated with arm movement to the hand position of a primate. The proposed architecture and the training algorithm are described in detail and numerical performance comparisons with alternative linear and nonlinear modeling approaches, including time-delay neural networks and recursive multilayer perceptrons, are presented. This new strategy allows training the local linear models using normalized LMS and using a relatively smaller nonlinear network to efficiently combine the predictions of the linear experts. This leads to savings in computational requirements, while the performance is still similar to a large fully nonlinear network. PMID- 12850046 TI - Stochastic error whitening algorithm for linear filter estimation with noisy data. AB - Mean squared error (MSE) has been the most widely used tool to solve the linear filter estimation or system identification problem. However, MSE gives biased results when the input signals are noisy. This paper presents a novel stochastic gradient algorithm based on the recently proposed error whitening criterion (EWC) to tackle the problem of linear filter estimation in the presence of additive white disturbances. We will briefly motivate the theory behind the new criterion and derive an online stochastic gradient algorithm. Convergence proof of the stochastic gradient algorithm is derived making mild assumptions. Further, we will propose some extensions to the stochastic gradient algorithm to ensure faster, step-size independent convergence. We will perform extensive simulations and compare the results with MSE as well as total-least squares in a parameter estimation problem. The stochastic EWC algorithm has many potential applications. We will use this in designing robust inverse controllers with noisy data. PMID- 12850047 TI - New internal optimal neurocontrol for a series FACTS device in a power transmission line. AB - In this paper, the proportional-integral (PI) based conventional internal controller (CONVC) of a power electronic based series compensator in an electric power system, is replaced by a nonlinear optimal controller based on the dual heuristic programming (DHP) optimization algorithm. The performance of the CONVC is compared with that of the DHP controller with respect to damping low frequency oscillations. Simulation results using the PSCAD/EMTDC software package are presented. PMID- 12850048 TI - Design of an adaptive neural network based power system stabilizer. AB - Power system stabilizers (PSS) are used to generate supplementary control signals for the excitation system in order to damp the low frequency power system oscillations. To overcome the drawbacks of conventional PSS (CPSS), numerous techniques have been proposed in the literature. Based on the analysis of existing techniques, this paper presents an indirect adaptive neural network based power system stabilizer (IDNC) design. The proposed IDNC consists of a neuro-controller, which is used to generate a supplementary control signal to the excitation system, and a neuro-identifier, which is used to model the dynamics of the power system and to adapt the neuro-controller parameters. The proposed method has the features of a simple structure, adaptivity and fast response. The proposed IDNC is evaluated on a single machine infinite bus power system under different operating conditions and disturbances to demonstrate its effectiveness and robustness. PMID- 12850049 TI - On neural network techniques in the secure management of communication systems through improving and quality assessing pseudorandom stream generators. AB - Random components play an especially important role in the management of secure communication systems, with emphasis on the key management of cryptographic protocols. For this reason, the existence of strong pseudo random number generators is highly required. This paper presents novel techniques, which rely on Artificial Neural Network (ANN) architectures, to strengthen traditional generators such as IDEA and ANSI X.9 based on 3DES and IDEA. Additionally, this paper proposes a non-linear test method for the quality assessment of the required non-predictability property, which relies on feedforward neural networks. This non-predictability test method along with commonly used empirical tests based on statistics is proposed as a methodology for quality assessing strong pseudorandom stream generators. By means of this methodology, traditional and Neural Network based pseudorandom stream generators are evaluated. The results show that the proposed generators behave significantly better than the traditional ones, in particular, in terms of non-predictability. PMID- 12850050 TI - Multimedia authenticity protection with ICA watermarking and digital bacteria vaccination. AB - We propose the application of independent component analysis (ICA), via unsupervised neural networks, to authenticity protection for multimedia products. We give an overview of the current state of multimedia authenticity protection, including the requirements of various multimedia applications, current approaches to the problem, and the robustness of the approaches. For watermark security, a covert independent-component watermarking signal can serve as a 'vaccination' against a dormant digital 'bacteria' protecting the multimedia data. An unauthorized removal of the watermark triggers the bacteria payload, which then degrades the quality of the unauthorized data. We argue that such digital bacteria meet all the established requirements for beneficial virus-like programs, and their payload would merely affect pirated media. We show how these new approaches contribute to a flexible, robust, and secure system for protecting the authenticity of multimedia products. PMID- 12850051 TI - Interpolation processes in the visual perception of objects. AB - Visual perception of objects depends on segmentation and grouping processes that act on fragmentary input. This paper gives a brief overview of these processes. A simple geometry accounting for contour interpolation is described, and its applications to 2D, 3D, and spatiotemporal object interpolation processes are considered. A method is described for distinguishing interpolation based on this geometry from more global or top-down influences. Results suggest a separation between interpolation based on relatively local stimulus relations, which give rise to precise boundary representations, and processes involving recognition from partial information, which do not. Aspects of the model-especially the unified treatment of illusory and occluded objects-raise questions about the nature of seeing. Although it is often believed that illusory objects are perceived, while occluded objects are inferred, I suggest that both research and theory converge in supporting a more unified account. Illusory and occluded contours and surfaces do not divide into the real, the perceived, and the inferred, but are all represented, and in key respects, derive from identical perceptual processes. PMID- 12850052 TI - Laminar cortical dynamics of visual form perception. PMID- 12850053 TI - Moving objects appear to slow down at low contrasts. AB - Moving cars give the illusion of slowing down in foggy conditions, because low contrast reduces perceived speed. A grey square that drifts horizontally across a surround of black and white vertical stripes appears to stop and start as it crosses each stripe, because its contrast keeps changing. A moving square whose vertical and horizontal edges have different contrasts will show illusory distortions in perceived direction. Contrast also affects the apparent amplitude and salience of back-and-forth apparent motion. Finally, a line of black and white dots on a grey surround moves in illusory directions, because of a mismatch in the contrasts along and across the dotted line. Thus, motion signals in the early parts of the visual system are profoundly altered by stimulus luminance and contrast. This suggests that motion is coded by the relative firing rates of neural channels tuned to fast and slow motion, with contrast-dependence being a motion analog of the Bezold-Brucke hue shift. PMID- 12850054 TI - Neural models of motion integration and segmentation. AB - A neural model is developed of how motion integration and segmentation processes compute global motion percepts. Figure-ground properties, such as occlusion, influence which motion signals determine the percept. For visible apertures, a line's extrinsic terminators do not specify true line motion. For invisible apertures, a line's intrinsic terminators create veridical feature tracking signals, which are amplified before they propagate across space and are integrated with ambiguous motion signals within line interiors. This integration process is the result of several processing stages: directional transient cells respond to image transients and input to a directional short-range filter that selectively boosts feature tracking signals. Competitive interactions further boost feature tracking signals and create speed-selective receptive fields. A long-range filter gives rise to true directional cells by pooling signals over multiple orientations and opposite contrast polarities. A distributed population code of speed tuning realizes a size-speed correlation, whereby activations of multiple spatially short-range filters of different sizes are transformed into speed-tuned cell responses. These mechanisms use transient cell responses, output thresholds that covary with filter size, and competition. The model reproduces empirically derived speed discrimination curves and simulates data showing how visual speed perception and discrimination are affected by stimulus contrast. PMID- 12850056 TI - Intracellular signaling pathways involved in mediating the effects of lead on the transcription factor Sp1. AB - It has been well established that exposure to Pb during critical periods of brain development results in both cognitive and behavioral deficits. Although the mechanism by which Pb induces developmental neurotoxicity is unknown, it may involve alterations in transcription of genes that are essential for growth and differentiation. Recent studies reveal that Pb interferes with growth and differentiation by acting on the transcription factor Sp1. Pb-induced changes in the activity of Sp1 may be consequent to alterations in intermediates in signal transduction pathways. This study examines both in vivo and in vitro the role of signaling factors in mediating the effects of Pb on Sp1 DNA-binding. Hippocampal developmental profiles of Sp1 DNA-binding, PKC, and MAPK protein levels were monitored in Pb-exposed rats. Pb exposure resulted in an induction of Sp1 DNA binding during PND 5-10 followed by a subsequent decline on PND 15-20. The protein expression profiles for PKCalpha and MAPK followed a relatively similar pattern. To examine the interdependence between Sp1 DNA-binding, PKCalpha, and MAPK, PC12 cells were exposed to Pb and/or NGF. Pb or NGF exposure increased Sp1 DNA-binding. Addition of the PKC inhibitor (staurosporine) diminished NGF and Pb induced Sp1 DNA-binding, while the MAPK inhibitor (PD 98059), completely abolished both basal and induced Sp1 DNA-binding. These findings demonstrate that Sp1 DNA-binding is regulated by PKC and MAPK, which may serve as mediators through which Pb may indirectly modulate Sp1 DNA-binding. PMID- 12850057 TI - Sex differences in GABA(A)ergic system in rat substantia nigra pars reticulata. AB - The substantia nigra pars reticulata (SNR) is involved in the control of movement disorders including seizures through its GABAergic neurons. Microinfusions of muscimol (a GABA(A) receptor agonist) produce specific effects on seizures depending on sex, infusion site (SNR(anterior) or SNR(posterior)) and age. To assess whether these effects are due to sex differences in GABAergic indices within the SNR we analyzed the expression of alpha(1) subunit mRNA of the GABA(A) receptor and the levels of GABA immunoreactivity (IR) of male and female rats at postnatal day 15 (PN15) and PN30. In each age, within the same SNR region, expression of alpha(1) subunit mRNA and intensity of GABA IR per neuron was higher in females compared to males. At PN15, in both sexes, there were no regional differences in expression of alpha(1) subunit mRNA and intensity of GABA IR. However, at PN30 in both sexes, expression of alpha(1) subunit mRNA and intensity of GABA IR per cell was higher in SNR(anterior) than in SNR(posterior). These results demonstrate that expression of alpha(1) subunit mRNA for GABA(A) receptor and levels of GABA IR in the SNR are sex- and site-specific, which may contribute to sex-, regional- and age-related differences in the expression of movement disorders and seizures. PMID- 12850058 TI - Sargaquinoic acid promotes neurite outgrowth via protein kinase A and MAP kinases mediated signaling pathways in PC12D cells. AB - We previously isolated a nerve growth factor (NGF)-dependent neurite outgrowth promoting substance MC14 (sargaquinoic acid) from a marine brown alga, Sargassum macrocarpum. In the present study, the NGF-potentiating activity of MC14 to neural differentiation of PC12D cells was investigated in detail. The treatment of cells with 3 microg/ml MC14 in the presence of 1.25-100 ng/ml NGF markedly enhanced the proportion of neurite-bearing cells compared with the NGF-only controls. In addition, MC14 significantly elevated the NGF-induced specific acetylcholinesterase (AchE) activity in PC12D cells, suggesting that MC14 could morphologically and biochemically promote the differentiation of PC12D cells. The mechanism of action of MC14 was further investigated by pharmacological inhibition of several intracellular signaling molecules. Results indicated that the neurite outgrowth promoting activity of MC14 was almost completely blocked by 10 microM PD98059, suggesting that a TrkA-dependent MAP kinases-mediated signaling pathway may play a crucial role in modulating the effect of MC14. Besides, the MC14-enhanced neurite outgrowth was substantially suppressed by the pretreatment with 10 ng/ml protein kinase A (PKA) inhibitor, demonstrating that the adenylate cyclase-PKA signaling cascade was also involved in the action of MC14. In contrast, a PKC inhibitor chelerythrine chloride did not inhibit the neurite outgrowth promoting activity of MC14. Altogether, these results demonstrate that MC14 enhances the neurite outgrowth by cooperating at least two separated signaling pathways, a TrkA-MAP kinases pathway and an adenylate cyclase PKA pathway, in PC12D cells. PMID- 12850059 TI - Magnetic field exposure during gestation: pineal and cerebral cortex serotonin in the rat. AB - Extremely low frequency (ELF) magnetic fields seem to have a reproducible influence on cells in transitional states, such as cells during the embryonic and early postnatal periods. Intense and continuous serotonergic synaptic growth is present during the first 2 weeks of postnatal development, paralleled by 5-HT content in the brain, so, the effect of ELF on 5-HT content in the cerebral cortex and pineal gland was determined in growing rats exposed during pregnancy, and in normal controls. The results showed a significant 5-HT increase at birth, 15 and 21 days, in the cerebral cortex. No differences were found in the pineal gland. These short MF exposures had a long term effect on cerebral cortex 5-HT, possibly starting since the fetal period. The relevance of the present findings are discussed as related to the serotonin trophic role on the brain cortex. PMID- 12850060 TI - A novel dynamin III isoform is up-regulated in the central nervous system in hypothyroidism. AB - Hypothyroidism in early postnatal development leads to abnormal CNS development that may be controlled in part at the level of gene transcription. Comparing the expression of euthyroid (EuT) and hypothyroid (HypoT) rat brain mRNAs by differential display PCR (ddPCR), we identified a novel dynamin III mRNA that was up-regulated in the hypothyroid state. Northern analysis of brain mRNA using a probe from the dynamin III open reading frame (ORF) revealed two transcripts of 3.0 and 7.2kb size. The 3.0 kb transcript was observed in testis and brain, but not liver or lung RNA. In the brain the 3.0 kb transcript increased from 25 to 57% of adult (Ad) levels from postnatal day (p) p2-p15, but was not significantly regulated by thyroid hormone status. In contrast, the more abundant 7.2 kb transcript increased from 16.8 to 48.0% of adult levels from p2 to p15 in euthyroid rat pups but from 54.0% of adult levels at p2 to 97.9% of adult levels by p15 in hypothyroid pups. Overlapping cDNA clones from a rat brain cDNA library defined the 7.2kb mRNA, which consisted of the complete ORF, containing a four amino acid insert at the end of the pleckstrin homology domain (PHD), and two unique 3'-flanking regions, that are likely derived from alternative processing. Thus, the 7.2 kb dynamin III transcript is brain-specific and selectively regulated by thyroid hormone status. The data suggest that the regulation of dynamin III by altered thyroid hormone status may affect synaptogenesis in the CNS through dynamin's essential roles in synaptic vesicle and receptor recycling, neurotransmitter reuptake, and growth factor receptor signaling. PMID- 12850061 TI - Effect of Cuscuta chinensis glycoside on the neuronal differentiation of rat pheochromocytoma PC12 cells. AB - Exposure of rat pheochromocytoma PC12 cells to Cuscuta chinensis glycoside induced neuronal differentiation with resulting outgrowth of neurites and increase of acetylcholinesterase activity. A specific inhibitor of mitogen activated protein kinase (MAPK) kinase, PD98059, prevented this effect of C. chinensis on PC12 cells. These results suggested that C. chinensis glycoside induced neuronal differentiation in PC12 cells linked to the mitogen-activated protein kinase signaling cascade. PMID- 12850062 TI - Serotonergic receptors in the brain of in utero undernourished rats. AB - In this study, we report that 5-HT(1A) receptors are already present in fractions of axonal growth cones, from the normal rat fetal brain (E-17). Also, in utero undernourished (UN) rat pups at birth show a noteworthy enhancement in the B(max) of [3H]5-hydroxytryptamine (5-HT) and [3H]8-hydroxy-(2-N,N-dipropilamin)-tetralin (([3H])8-OH-DPAT), in the brainstem and cerebral cortex up to the second week after birth. Afterwards, there is a significant decrease in the binding of these ligands. [125I]Cyanopindolo binding in the cerebral cortex only showed a decrease in the same period. An elevation of brain serotonin in both regions was also present. These findings together, suggest that the mechanisms of regulation of serotonergic receptors' expression during the period studied, may not depend on the amount of neurotransmitter in the synaptic cleft, because in the early UN brain it would be expected only a lower receptor's density due to the chronic serotonin increase. On this basis, we propose that developmental activation of brain serotonin biosynthesis observed in early UN animals may disrupt the mechanism regulating the expression of 5-HT receptors during development. PMID- 12850063 TI - Reducing injuries among Native Americans: five cost-outcome analyses. AB - This paper presents cost-outcome analyses of five injury prevention efforts in Native American jurisdictions: a safety-belt program, a streetlight project, a livestock control project, a drowning prevention program, and a suicide prevention and intervention program. Pre- and post-intervention data were analyzed to estimate projects' impact on injury reduction. Projects' costs were amortized over the time period covered by the evaluation or over the useful life of physical capital invested. Projects' savings were calculated based on estimated reduction in medical and public program expenses, on estimated decrease in lost productivity, and on estimated quality adjusted life years saved.All projects yielded positive benefit-cost ratios. The net cost per quality adjusted life years was less than zero (i.e. the monetary savings exceeded project costs) for all but one of the projects. PMID- 12850064 TI - Consideration of driver home county prohibition and alcohol-related vehicle crashes. AB - This study examines the characteristics of alcohol-related crashes in wet versus dry counties in the state of Kentucky, USA and incorporates the location of driver residences through use of geographic information system (GIS) analysis. Between 1991 and 1997, 39344 alcohol-related crashes by Kentucky residents on Kentucky State roads were reported. The location of the crash and the home ZIP code from the driver's address were used to consider distance from home in the GIS. Analysis of the crash data revealed that a similar proportion of crashes in wet and dry counties are alcohol-related but that a higher proportion of dry counties residents are involved in an alcohol-related crash. However, when the distance from home variable is considered, several results suggest that dry county residents may be driving further when consuming alcohol. In part due to the rural nature of dry counties, drivers from dry counties have both alcohol related and non-alcohol related crashes farther from their homes than residents from wet counties. Alcohol-related crashes by dry county residents in wet counties are the greatest average distance from home while crashes by wet county residents in wet counties are the smallest average distance. Drivers from dry counties over 21 years of age have alcohol-related crashes significantly farther from home than those under 21 who would not legally be admitted to drinking establishments in the wet counties. Furthermore, residents from dry counties that do not border wet counties have alcohol-related crashes on average farther from home than the border county residents. These last three results provide circumstantial evidence that some dry county drivers may be driving to wet counties to consume alcohol thus increasing impaired driving exposure. In conclusion, by considering crash location and driver residence, these findings indicate that county-level prohibition is not necessarily effective in improving highway safety. PMID- 12850065 TI - Wireless telephones and the risk of road crashes. AB - In light of the rapidly increasing development of the cell phone market, the use of such equipment while driving raises the question of whether it is associated with an increased accident risk; and if so, what is its magnitude. This research is an epidemiological study on two large cohorts, namely users and non-users of cell phones, with the objective of verifying whether an association exists between cell phone use and road crashes, separating those with injuries. The Societe de l'Assurance Automobile du Quebec (SAAQ) mailed a questionnaire and letter of consent to 175000 licence holders for passenger vehicles. The questionnaire asked about exposure to risk, driving habits, opinions about activities likely to be detrimental to driving and accidents within the last 24 months. For cell phone users, questions pertaining to the use of the telephone were added. We received 36078 completed questionnaires, with a signed letter of consent. Four wireless phone companies provided the files on cell phone activity, and the SAAQ the files for 4 years of drivers' records and police reports. The three data sources were merged using an anonymized identification number. The statistical methods include logistic-normal regression models to estimate the strength of the links between the explanatory variables and crashes. The relative risk of all accidents and of accidents with injuries is higher for users of cell phones than for non-users. The relative risks (RR) for injury collisions and also for all collisions is 38% higher for men and women cell phone users. These risks diminish to 1.1 for men and 1.2 for women if other variables, such as the kilometres driven and driving habits are incorporated into the models. Similar results hold for several sub-groups. The most significant finding is a dose response relationship between the frequency of cell phone use, and crash risks. The adjusted relative risks for heavy users are at least two compared to those making minimal use of cell phones; the latter show similar collision rates as do the non-users. PMID- 12850066 TI - Behavioural characteristics and accidents: findings from the Health Survey for England, 1997. AB - This study analysed the relationship between major and minor accidents, and major accidents involving a moving vehicle, and behavioural and emotional factors in children, aged 4-15 years, using the Strengths and Difficulties Questionnaire (SDQ), and adjusting for demographic, socio-economic and family type factors. Data from a large representative national sample of about 6000 children were analysed using simple and multiple logistic regression. The analysis shows that the prevalence of SDQ scales, such as hyperactivity and conduct disorder were significantly higher in boys, lower social classes and step- and single-parent families. After adjusting for the demographic, socio-economic and family type factors, children who scored borderline or high for hyperactivity were almost two times more likely to report having major accidents. Children who scored high for hyperactivity and emotional symptoms were one and a half times more likely to report having minor accidents. For major accidents involving moving vehicles, the relationships with the behavioural and emotional factors were generally stronger than for major accidents in general. Hyperactivity, in particular, was significantly associated with the occurrence of major and minor accidents, and major accidents involving moving vehicles. The behavioural risk factors were significantly more common in the lower social classes, families receiving benefits and step- and single-parent families. PMID- 12850067 TI - Identifying factors that predict persistent driving after drinking, unsafe driving after drinking, and driving after using cannabis among young adults. AB - The main aim of this study was to identify adolescent/young adulthood factors that predicted persistent driving after drinking, persistent unsafe driving after drinking, and persistent cannabis use and driving among young adults. It was a longitudinal study of a birth cohort (n=933, 474 males and 459 females) and was based on data collected at ages 15, 18, 21 and 26 years. At each of these ages members of the cohort attended the research unit for a personal interview by a trained interviewer, using a standardised questionnaire. For this study, the data for the outcome measures (persistent driving after drinking, persistent unsafe driving after drinking, and persistent driving after using cannabis) were obtained at ages 21 and 26 years. The main explanatory measures were collected at ages 15, 18, 21 years and included demographic factors (academic qualifications, employment, parenting); personality measures; mental health measures (substance use, cannabis dependence, alcohol dependence, depression); anti-social behaviour (juvenile arrest, aggressive behaviour, court convictions); early driving behaviour and experiences (car and motorcycle licences, traffic crashes). The analyses were conducted by gender. The results showed that females who persisted in driving after drinking (13%, n=61) were more likely than the others to have a motorcycle licence at 18. The males who persisted in driving after drinking (28%, n=135) were more likely than the other males to have some school academic qualifications and to be employed at age 26. Compared to the other males, those who persisted in unsafe driving after drinking (4%, n=17) were more likely to be aggressive at 18 and alcohol dependent at 21. Only six (1%) females persisted in unsafe driving after drinking so regression analyses were not conducted for this group. For persistent driving after using cannabis, the univariate analyses showed that females who persisted with this behaviour tended to have high substance use at 18, cannabis dependence at 21, police contact as a juvenile, and to be a parent at 21. For this group, because of the small numbers (3%, n=13) multivariate analyses were not appropriate. For the males who persisted in driving after using cannabis (14%, n=68) a wide range of variables were significant at the univariate stage. The multivariate analysis showed that the most important factors were dependence on cannabis at 21, at least one traffic conviction before 21, a non traffic conviction before 18, and low constraint at 18. CONCLUSION: These results show different characteristics were associated with persistence in each of these outcome behaviours. This indicates that different approaches would be required if intervention programmes were to be developed to target these behaviours. PMID- 12850068 TI - Factors associated with the likelihood of injury resulting from collisions between four-wheel drive vehicles and passenger cars. AB - The specific effects of vehicular type on the likelihood of an injury occurring are relatively unexplored. This study sought to assess the relative risk of injury to occupants of four-wheel drive vehicles and their counterparts in passenger cars. Data for 1143 occupants from all of the 454 crashes in Oklahoma, in 1995 that involved a four-wheel drive vehicle were used. Multiple logistic regression analysis determined the association between potential predictive factors and vehicular injury. Odds ratios revealed occupancy in a passenger car to be a major predictor of the likelihood of injury. Other factors include the driver being female, driving too fast, travel on curved or level roadways, and being hit laterally or from the rear. PMID- 12850069 TI - Changes in collision rates among novice drivers during the first months of driving. AB - As a group, young drivers have crash rates that far exceed those of older, experienced drivers. But even among teenagers there are age-related differences; crash rates decline consistently and dramatically with each yearly increase in age. A more precise understanding of how crash rates decline within the teenage group might provide insights into when experience is most influential. To address this issue, this study examines month-to-month changes in collisions among new drivers. It was found that crash rates drop most dramatically during the first 6 months of driving. Involvement in certain types of crashes-e.g. run-off-the-road, single-vehicle, night, weekend-declines more rapidly. The findings suggest that novices improve their driving in a relatively short period of time. A graduated driver licensing system is identified as an effective method for ensuring that this development takes place in a more forgiving environment. PMID- 12850070 TI - A disaggregate approach to crash rate analysis. AB - This paper presents a disaggregate approach to crash rate analysis. Enumerating crash rates on a per trip-kilometer basis, the proposed method removes the linearity assumption inherent in the conventional quotient indicator of accidents per unit travel distance. The approach involves combining two disparate datasets on a geographic information systems (GIS) platform by matching accident records to a defined travel corridor. As an illustration of the methodology, travel information from the Victorian Activity and Travel Survey (VATS) and accident records contained in CrashStat were used to estimate the crash rates of Melbourne residents in different age-sex groups according to time of the day and day of the week. The results show a polynomial function of a cubic order when crash rates are plotted against age group, which contrasts distinctly with the U-shape curve generated by using the conventional aggregate quotient approach. Owing to the validity of the many assumptions adopted in the computation, this study does not claim that the results obtained are conclusive. The methodology, however, is seen as providing a framework upon which future crash risk measures could be based as the use of spatial tracking devises become prevalent in travel surveys. PMID- 12850071 TI - Firearms in US homes as a risk factor for unintentional gunshot fatality. AB - This study used national data and a matched case-control design to estimate the relative risk of death by an unintentional gunshot associated with having firearms in the home. A sample of adults who died in the United States in 1993 from unintentional gunshot injuries was drawn from the National Mortality Followback Survey (NMFS) (n=84). Twenty controls were sought for each case from the 1994 National Health Interview Survey (NHIS) and matched to the cases by sex, age group, race, and region of residence (n=1451). Subjects were classified as having or not having guns in the home based interview responses. The relative risk of death by an unintentional gunshot injury, comparing subjects living in homes with and without guns, was 3.7 (95% confidence interval (CI)=1.9-7.2). Adjustment for covariates resulted in little change in the effect estimates. There was evidence of a dose-response effect: compared to subjects living in homes with no guns, the relative risk was 3.4 (95% CI=1.5-7.6) among subjects with one gun and 3.9 (95% CI=2.0-7.8) among subjects with multiple guns in the home. Having handguns in the home was associated with the largest effect estimates. Tests of homogeneity showed that the effect estimates did not vary significantly across categories of the matching variables. Firearms in the home appear to be a risk factor for unintentional gunshot fatality among adults. The magnitude of the observed effect estimates should be compared with those from additional studies. PMID- 12850072 TI - Analysis of traffic accidents at urban intersections in Riyadh. AB - Previous studies have shown that intersection-related accidents account for about 50% of all accidents registered annually in Riyadh, the capital of the Kingdom of Saudi Arabia (KSA). More than half of these accidents are classified as severe. In this study, an attempt was made to investigate traffic accidents that occurred at both intersections and non-intersection sites. The goal was to analyze the nature of such accidents to determine their characteristics so that remedies could be sought or at least future research could be suggested. For this purpose, a sample of 1774 reported accidents was collected in a systematic random manner for the period 1996-1998 (651 severe accidents (accidents resulting in at least one personal injury or fatality) and 1123 property-damage-only (PDO) accidents). Conditional probability and contingency table analyses were used to make inferences from the data. The study found that improper driving behavior is the primary cause of accidents at signalized urban intersections in Riyadh; running a red light and failing to yield are the primary contributing causes. The analysis indicates that there is an urgent need to review existing intersection geometry along with the traffic control devices installed at these sites. In addition, public education campaigns and law enforcement strategies are urgently needed. PMID- 12850073 TI - Accident involvement among learner drivers--an analysis of the consequences of supervised practice. AB - It is a well-known fact that experience is important for safe driving. Previously, this presented a problem since experience was mostly gained during the most dangerous period of driving-the first years with a licence. In many countries, this "experience paradox" has been addressed by providing increased opportunities to gain experience through supervised practice. One question, however, which still needs to be answered is what has been lost and what has been gained through supervised practice. Does this method lead to fewer accidents after licensing and/or has the number of accidents in driving practice increased? There were three aims in the study. The first was to calculate the size of the accident problem in terms of the number of accidents, health risk and accident risk during practising. The second aim was to evaluate the solution of the "experience paradox" that supervised practice suggests by calculating the costs in terms of accidents during driving practice and the benefits in terms of reduced accident involvement after obtaining a licence. The third aim was to analyse conflict types that occur during driving practice. National register data on licence holders and police-reported injury accidents and self-reported exposure were used. The results show that during the period 1994-2000, 444 driving practice injury accidents were registered, compared to 13657 accidents during the first 2 years with a licence. The health risk during the period after licensing was 33 times higher and the accident risk 10 times higher than the corresponding risk during practice. The cost-benefit analysis showed that the benefits in terms of accident reduction after licensing were 30 times higher than the costs in terms of driving practice accidents. It is recommended that measures to reduce such accidents should focus on better education of the lay instructor, but not on introducing measures to reduce the amount of lay-instructed practice. PMID- 12850074 TI - Occupant deaths in large truck crashes in the United States: 25 years of experience. AB - There is public concern about the magnitude of the problem of large truck crashes in the US. Fatalities in large truck crashes have not declined much; however, more large trucks are driving more miles than ever before while fatalities per mile driven have dropped substantially. This study examined how the public health burden of large truck crashes versus the risk per unit of travel has changed over 25 years. The present study focused on the US vehicle occupants in fatal crashes involving a large truck during 1975-1999. Occupant fatalities per 100000 population, per 10000 licensed drivers, per 10000 registered trucks and per 100 million vehicle-miles of travel (VMT) were calculated to determine trends in occupant deaths in large truck crashes. In 1999, large truck crashes resulted in 3916 occupant deaths in passenger vehicles and 747 in large trucks. Passenger vehicle occupant deaths in large truck crashes per 100000 population have increased somewhat since 1975 (1.28 in 1975 and 1.44 in 1999). There have been appreciable declines in occupant deaths per truck VMT since 1975, but the percentage reduction has been greater for occupants of large trucks (67%) than for passenger vehicle occupants (43%). However, truck drivers are at elevated risk of dying relative to their numbers in the workforce. Overall large truck involvements in fatal crashes per truck VMT decreased more than passenger vehicle involvements per passenger VMT (PVMT; 68% versus 33% decreases for single-vehicle crashes and 43% versus 23% for multiple-vehicle crashes). Large truck involvement in fatal crashes has dropped substantially when measured per unit of travel, but the public health burden of large truck crashes, as measured by deaths per 100000 population, has not improved over time because of the large increase in truck mileage. Research is needed on measures to better protect both occupants of large trucks and passenger vehicle occupants colliding with them. PMID- 12850075 TI - Assessing the validity of road safety evaluation studies by analysing causal chains. AB - This paper discusses how the validity of road safety evaluation studies can be assessed by analysing causal chains. A causal chain denotes the path through which a road safety measure influences the number of accidents. Two cases are examined. One involves chemical de-icing of roads (salting). The intended causal chain of this measure is: spread of salt --> removal of snow and ice from the road surface --> improved friction --> shorter stopping distance --> fewer accidents. A Norwegian study that evaluated the effects of salting on accident rate provides information that describes this causal chain. This information indicates that the study overestimated the effect of salting on accident rate, and suggests that this estimate is influenced by confounding variables the study did not control for. The other case involves a traffic club for children. The intended causal chain in this study was: join the club --> improve knowledge --> improve behaviour --> reduce accident rate. In this case, results are rather messy, which suggests that the observed difference in accident rate between members and non-members of the traffic club is not primarily attributable to membership in the club. The two cases show that by analysing causal chains, one may uncover confounding factors that were not adequately controlled in a study. Lack of control for confounding factors remains the most serious threat to the validity of road safety evaluation studies. PMID- 12850076 TI - Traffic sense--which factors influence the skill to predict the development of traffic scenes? AB - A study was conducted to evaluate the skill to predict the development of traffic situations. A stop-controlled intersection was filmed over several days, and 12 scenes with varying traffic complexity were selected. In half of the scenes, the traffic rules were violated, in half of the scenes, the rules were observed. A total of 36 participants were asked to watch the scenes and predict how the scene would most likely develop in the 2s after the film was paused. Additionally, the participants rated how certain they were about their prediction, and how complex and dangerous they assessed the scenes to be. With the method used here, experienced drivers were not found to make more correct predictions of situational development, and no difference in skill to predict could be found between genders. Nevertheless, more experienced drivers were more certain in their judgements and evaluated the situations on average as less complex and dangerous than did less experienced drivers. Scenes in which the traffic rules were violated were more difficult to predict correctly. The scenes in which the participants predicted violations were rated as more complex and dangerous. It is concluded that the low-cost method used here is more useful for examining which scenes are generally easy or difficult to predict and how they are experienced subjectively than to investigate differences in performance for different driver categories. PMID- 12850077 TI - Evidence for an early onset of endogenous alcohol production in bodies recovered from the water: implications for studying alcohol and drowning. AB - Endogenous alcohol production can increase the blood alcohol concentration (BAC) of drowning victims following submersion and confound epidemiological studies of the role of alcohol. This study seeks to determine how soon after a drowning death a victim's BAC is influenced by post-mortem alcohol production. The drop in mean lung weight that occurs over time in the water was hypothesized to serve as a proxy for the time course of decomposition, and thus provide an empirical measure to determine how soon after death to first suspect endogenous alcohol. The autopsy lung weights of 562 previously healthy males who drowned were compared across six submersion time groups (0-11.9, 12-23.9, 24-47.9, 48-95.9, 96 167.9 and >or=168 h) and two times of year (winter and non-winter). The hypothesis that a drop in lung weight is sensitive to the time course of decomposition was supported by (1). a statistically significant drop in mean lung weight that occurred 12-23.9 h post-submersion in the non-winter months, but not until 96-167.9 h in the colder winter months; and (2). a significant drop in lung weight was not observed in the group of cases with zero BAC. With a parallel finding that an increase in the proportion of cases with a positive BAC first occurred at the 12-23.9 h submersion group during the warmer non-winter months, we concluded that production of alcohol can occur in bodies recovered from the water as early as 12 h after death. Because excluding drownings with submersion durations greater than 12 h would exclude almost half of our cases from epidemiological studies of alcohol and drowning, additional evidence from the forensic literature was used to develop an adjustment procedure to account for endogenous alcohol production for submersion times of up to 1 week. PMID- 12850078 TI - Behavioural observations of adult-child pairs at pedestrian crossings. AB - Pedestrian accidents are a serious health risk to children in the UK and other Western countries. There remains a considerable amount to be learned about children's behaviour in real-traffic environments. The present study was intended to help fill this gap, by observing unobtrusively the behaviours of 123 adult child pairs as they crossed the road at pedestrian light-controlled crossings. Eight behaviours were coded, including whether or not the pair stopped at the kerb, waited for the light to change, and checked to ensure traffic flow had stopped. Results showed that the adults observed provided reasonably good models of pedestrian behaviour, but that they rarely treated the crossing event as an opportunity to teach children explicitly about road safety. The only gender difference to emerge revealed that adults were more likely to hold girls' hands than boys' hands. No differences were observed in relation to (estimated) age of child. The implications of these findings for parental training programmes are explored. PMID- 12850079 TI - Studies of neck injury criteria based on existing biomechanical test data. AB - The mechanisms of injury of the human neck are potentially related to the forces and bending moments acting on the spinal column. An injury criterion that combines the effects of the time history of force and moment, N(ij), has been adopted by the USA federal government for vehicle crash regulations. This paper presents two analytical approaches to evaluate the N(ij) injury criterion using previously published biomechanical data: a direct fit optimization analysis and a statistical analysis. The results show that no values used in the N(ij) criterion corresponding to compressive force, extension moment and flexion moment will optimally separate the injury and non-injury cases in the biomechanical data set. Only the tensile force allows identification of the injury risk. All indications are that increasing the weight given to moment decreases the accuracy of the prediction. PMID- 12850080 TI - An investigation of road crossing in a virtual environment. AB - The reported study employed a virtual reality (VR) system, using a head mounted display (HMD), to investigate road crossing behavior in children and young adults. Younger children (aged 5-9 years) made the greatest number of unsafe road crossings and the oldest participants (aged >19 years) the fewest. Overall performance was better (fewer unsafe road crossings) in uniform speed than uniform distance trials, consistent with previous research suggesting that pedestrians base road crossing decisions on inter-vehicle distance rather than vehicle speed. Results are discussed in terms of road crossing behavior and the use of VR simulations in the study of pedestrian behavior. PMID- 12850081 TI - Assessing the driving performance of older adult drivers: on-road versus simulated driving. AB - To validate a laboratory-based driving simulator in measuring on-road driving performance, 129 older adult drivers were assessed with both the simulator and an on-road test. The driving performance of the participants was gauged by appropriate and reliable age-specific assessment criteria, which were found to be negatively correlated with age. Using principal component analysis, two performance indices were developed from the criteria to represent the overall performance in simulated driving and the on-road assessment. There was significant positive association between the two indices, with the simulated driving performance index explaining over two-thirds of the variability of the on road driving performance index, after adjustment for age and gender of the drivers. The results supported the validity of the driving simulator and it is a safer and more economical method than the on-road testing to assess the driving performance of older adult drivers. PMID- 12850082 TI - Older drivers and road safety: the acceptability of a range of intervention measures. AB - A survey questionnaire was used to ascertain the views of 1932 UK drivers aged between 50 and 90 on a range of measures designed to promote safer driving among the elderly. Factor analysis of the items produced six factors, relating to statistically distinct types of measure. Differences in acceptability on the basis of age and sex were explored. Ratings of the effectiveness of each measure revealed little consensus about which measures would be most effective. Compulsory re-testing after a driving ban, a police power to require an assessment of the driving of anyone observed driving in a risky manner, and a requirement for opticians to report to the licensing authority any driver with sight problems likely to affect driving emerged as the individual measures offering the best combination of acceptability and perceived effectiveness. PMID- 12850083 TI - Small female head and neck interaction with a deploying side airbag. AB - OBJECTIVE: This paper presents dummy and cadaver experiments designed to investigate the injury potential of an out-of-position small female head and neck from a deploying side airbag. METHODS: Seat-mounted, thoracic-type, side airbags were selected for this study to represent those currently available on selected luxury automobiles. A computer simulation program was used to identify the worst case loading position for the small female head and neck. Once the initial position was identified, experiments were performed with the Hybrid III 5th percentile dummy and three small female cadavers, using three different inflators. RESULTS: Peak head center of gravity (CG) accelerations for the dummy ranged from 71x g to 154 x g, and were greater than cadaver values, which ranged from 68 x g to 103 x g. Peak neck tension as measured at the upper load cell of the dummy increased with inflator aggressivity from 992 to 1670N. A conservative modification of the US National Highway Traffic Safety Administration's (NHTSA's) N(ij) proposed neck injury criteria, which combines neck tension and bending, was used. All values were well below the 1.0 injury threshold for the dummy and suggested a very low possibility of neck injury. In agreement with this prediction, no injuries were observed. CONCLUSIONS: Even in a worst case position, small females are at low risk of head or neck injuries under loading from these thoracic-type airbags; however, injury risk increases with increasing inflator aggressivity. PMID- 12850084 TI - Restricting intersection visibility to reduce approach speeds. AB - This paper reports the field test of a visual restriction treatment for a rural intersection with a high rate of injury crashes. A human factors analysis of the asymmetric pattern of crashes at the site suggested that most of the crashes were the result of anticipatory decision-making occasioned by visual characteristics of the eastbound approach to the intersection. The field test examined the effectiveness of a visual restriction treatment directed at eliminating drivers' anticipatory decision-making. The treatment consisted of a hessian screen erected along the eastbound approach to the intersection beginning 125 m prior to intersection and ending 25 m prior to intersection. Over 2 days of testing, approximately 300 drivers' reactions at the intersection were observed and their responses to a brief survey recorded. The test indicated a 23% reduction in the 80th percentile and mean approach speeds and elimination of all approach speeds over 57 km/h following introduction of the treatment. Survey results showed that the treatment was visually acceptable to the majority of drivers using the intersection and did not affect its perceived safety. Follow-on analyses compared speed data before the treatment, and 2, 21, and 37 weeks after installation of the treatment. These analyses showed that approach speeds remained low; 30% lower than pre-treatment speeds for both the 80th percentile and the average approach speeds. Of perhaps the greatest significance, no crash resulting in serious injury or death has occurred at the intersection since installation of the treatment to the present time. PMID- 12850085 TI - The state-of-the-science and trends of brominated flame retardants in the environment: present knowledge and future directions. PMID- 12850086 TI - Environmental release and behavior of brominated flame retardants. AB - Recently, environmental problems relating to brominated flame retardants (BFRs) have become a matter of greater concern than ever before, because of the recent marked increase in levels of polybrominated diphenyl ethers (PBDEs) found in human milk in Sweden and North America. The question that arises is whether environmental levels of PBDEs and other BFRs will continue to increase, causing toxic effects to humans. In an attempt to elucidate the current state of the science of BFRs, we review the consumer demand for BFRs (mainly in Japan), the characteristics of waste flame-retarded products, sources of emission, environmental behavior, routes of exposure of humans, temporal trends, and thermal-breakdown products of BFRs. At present, flame-retarded consumer products manufactured 10-20 years ago, when PBDEs were frequently used, are being dumped. The possible major sources of emission of BFRs into the environment are effluent and flue gases from BFR factories and other facilities processing BFRs. With respect to the environmental behavior of BFRs, the lower brominated compounds are, on the whole, predicted to be more volatile, more water soluble, and more bioaccumulative than the higher brominated compounds. The most probable route for exposure of the general human population to PBDEs, especially the lower brominated congeners, is through the diet. The release of BFRs from consumer products treated with these compounds could also lead to human exposure. Temporal trends in PBDE levels in the environment and in humans worldwide seem to vary considerably, depending on the regions or country, with possible reflections of the historic and current use of PBDEs. The environment and the general human population are also exposed to the thermal-breakdown products of PBDEs, such as polybrominated and mixed brominated/chlorinated dibenzo-p-dioxins and dibenzofurans (PBDDs/DFs and mixed PXDDs/DFs). PMID- 12850087 TI - An overview of commercially used brominated flame retardants, their applications, their use patterns in different countries/regions and possible modes of release. AB - Brominated flame retardants (BFRs) are used in a variety of consumer products and several of those are produced in large quantities. These compounds have been detected in environmental samples, which can be attributed to the anthropogenic uses of these compounds. Brominated flame retardants are produced via direct bromination of organic molecules or via addition of bromine to alkenes; hence, an overview of the production and usage of bromine over the past three decades is covered. Production, application, and environmental occurrence of high production brominated flame retardants including Tetrabromobisphenol A, polybrominated biphenyls, Penta-, Octa-, Deca-brominated diphenyl ether (oxide) formulation and hexabromocyclododecane are discussed. PMID- 12850088 TI - Understanding levels and trends of BDE-47 in the UK and North America: an assessment of principal reservoirs and source inputs. AB - Despite increasing interest in the occurrence of Polybrominated diphenyl ethers (PBDEs) in the environment, there has been only limited effort expended to identify principal source input reservoirs and pathways into the environment. Taking a single congener BDE-47, an important component of the penta commercial product, we have estimated principal contemporary reservoirs and emissions in two regions with very different historical use patterns. In North America, production and use of the penta-product has been rising steadily for the last 20 years, whilst in the UK, use has been restricted over the last 10 years, and most recently, there has been an EU ban on use. Mass balance models have been applied using our contemporary emission estimates to predict environmental levels. Results highlight that contemporary emissions to air can account for contemporary air concentrations but cannot account for measured concentrations in principal reservoirs such as soils and sediments. Future efforts are needed to improve our knowledge of the stocks of PBDEs in use, and emission factors from those stocks, to better understand/predict past and likely future trends. PMID- 12850089 TI - Relevance of BFRs and thermal conditions on the formation pathways of brominated and brominated-chlorinated dibenzodioxins and dibenzofurans. AB - The widespread use of brominated flame-retarded products in the last two decades has resulted in an increasing presence of bromine in thermal processes such as waste combustion and accidental fires. Brominated and brominated-chlorinated dibenzodioxins and dibenzofurans (PBDDs/PBDFs, PXDDs/PXDFs) are micropollutants of concern arising from such processes. The present review aims to evaluate the relevance of these compound classes in actual thermal processes. Four categories of thermal processes are discussed in this respect according to their potential for PBDD/PBDF and PXDD/PXDF generation: thermal stress, pyrolysis/gasification, insufficient combustion conditions and controlled combustion conditions. Under thermal stress situations, as they may occur in production or recycling processes, PBDDs/PBDFs precursors like polybrominated diphenylethers (PBDE) can have a relevant potential for PBDD/PBDF formation via a simple elimination. Under insufficient combustion conditions as they are present in, e.g. accidental fires and uncontrolled burning as well as gasification/pyrolysis processes, considerable amounts of PBDDs/PBDFs can be formed from BFRs, preferably via the precursor pathway. In contrast, under controlled combustion conditions, BFRs and PBDDs/PBDFs can be destroyed with high efficiency. The relevance of de novo synthesis of PXDDs/PXDFs is discussed for this condition. Providing a basis for the understanding of PXDD/PXDF formation in actual thermal processes, the present paper also summarises the formation pathways of brominated and brominated chlorinated PXDDs/PXDFs from brominated flame retardants (BFRs) investigated during laboratory thermolysis experiments. Relevant mechanistic steps for PBDD/PBDF formation from brominated precursors are discussed including elimination reactions, condensation steps and debromination/hydrogenation reactions. In addition, chlorination/bromination and halogen exchange reactions are briefly discussed with respect for their relevance on the final distribution of PBDDs/PBDFs, mixed chlorinated PXDDs/PXDFs and PCDDs/PCDFs resulting from thermal processes. PMID- 12850090 TI - Formation of PBDD/F from flame-retarded plastic materials under thermal stress. AB - Thermal treatment of plastics flame retarded with brominated compounds often lead to the formation of PBDD/F, especially if polybrominated diphenyl ethers are used. By the presence of antimony(III) oxide, water and some other metals, the amounts of PBDD/F increase. Even during the production and recycling processes of plastics, PBDD/F can be found in considerable amounts. By the use of flame retardants, which are not direct precursors of PBDD/F, the amounts can significantly be decreased. An incomplete and uncontrolled incineration, occurring in fire accidents in private residences, can form high amounts of PBDD/F, which can endanger the environment and persons in contact with fire residues. Complete and controlled incineration of bromine-containing compounds as usually conducted in municipal waste incinerators, however, show no raised risk. An overview about the formation of PBDD/F primarily during combustion and also during production and recycling processes of flame-retarded plastic materials is given in this paper. PMID- 12850091 TI - Modelling the environmental fate of the polybrominated diphenyl ethers. AB - In response to growing alarm over the occurrence of polybrominated diphenyl ethers (PBDEs) in remote regions, this study considers their physical chemistry, environmental partitioning and considerations regarding potential for long-range atmospheric transport (LRAT). Internally consistent physical-chemical property data are presented for five representative congeners (PBDE-15, -28, -47, -99, 153) and used in a multimedia modelling approach. Results of the Level II model indicate that PBDEs will largely partition to organic carbon in soil and sediment and that their persistence will be strongly influenced by degradation rates in these media that are not well known. TaPL3 model estimates of their characteristic travel distance (CTD) suggest limited LRAT potential. The LRAT is also evaluated qualitatively, in terms of surface-air exchange behaviour. PBDEs are shown to be sensitive to seasonally and diurnally fluctuating temperatures. When vegetation is included in the model, 50% of the total mass of PBDE-47 deposited to vegetation returns to the atmosphere, suggesting that it may migrate through a series of deposition/volatilisation hops. Key data that needs to be identified in this evaluation include a better understanding of air-surface exchange, particularly to foliage, and measurements of degradation rates in soil, sediment and vegetation. PMID- 12850092 TI - Playing with fire: the global threat presented by brominated flame retardants justifies urgent substitution. AB - Few would now deny that the use of organobromine compounds to achieve fire retardancy in a diverse array of products and materials has led to contamination of the ecosphere on a widespread scale. This environmental prevalence and persistence of the brominated flame retardants, coupled with growing evidence of their potential for harm, present all too familiar parallels with the previous generation of persistent organic pollutants. Indeed, given the intrinsic properties of these brominated chemicals, the nature and extent of the current problem could well have been predicted in advance. The question is then whether we are prepared to let history repeat itself once more or to take precautionary action now to switch to more sustainable alternatives. The choice facing society is not between brominated flame retardants and unsafe products, but between fire safety leading to global contamination or fire safety achieved in less polluting ways. If we look beyond options for simple chemical-for-chemical substitution to alternative materials and designs, many of the solutions are already available. The remainder could undoubtedly be developed given the incentives to do so. However, a strong and clear policy approach, backed by legislative phase-outs within specified (and challenging) timeframes, will be necessary to break our current dependency on organobromine chemistry. This paper presents the justification for such an approach, reviews those initiatives already underway to replace brominated flame retardants and identifies pathways to the use of more sustainable products in the service of society. PMID- 12850093 TI - Determination of brominated flame retardants, with emphasis on polybrominated diphenyl ethers (PBDEs) in environmental and human samples--a review. AB - Analytical methods for the determination of brominated flame retardants (BFRs), with a special emphasis on polybrominated diphenyl ethers (PBDEs) are reviewed. A number of procedures, which can be applied to the analysis of PBDEs and polybrominated biphenyls (PBBs), and in some cases for hexabromocyclododecane (HBCD), in environmental and human samples are described. Because several BFRs, such as tetrabromobisphenol-A (TBBP-A), BDE 209 and, to some extent, HBCD, may require a different approach, specific advice on their analysis is given separately when needed. Sample pretreatment, extraction, cleanup and fractionation, injection techniques, chromatographic separation, detection methods, quality control and method validation are discussed. For each topic, an overview is given of the current status of the field and recommendations for an appropriate analytical approach are presented. PMID- 12850094 TI - Levels and trends of polybrominated diphenylethers and other brominated flame retardants in wildlife. AB - In this paper, we review the available data for polybrominated diphenylethers (PBDEs) and other flame retardants in wildlife, with the exception of fishes from Europe and North America which are covered in more detail elsewhere. More data are available for PBDEs than for other compounds, and these show that some of these compounds have become widely distributed in the environment, being found in samples from Europe, Australia, Azerbaijan, North America and the Arctic. Most available data relate to birds and their eggs and marine mammals, but the results of two food web studies are also included. The detection of PBDEs in pelagic marine mammals which feed in deep offshore waters, including baleen whales, indicate that these compounds have found their way into deep-water, oceanic food webs as well as the coastal/shallow sea examples described in detail. In the North Sea study, the most marked increase in lipid-normalised concentrations of six BDE congeners occurred during transfer from predatory fish to marine mammals. In the St. Lawrence Estuary study, marked differences in the ratios observed between species suggested that some fish species may be able to metabolise BDE99.A number of time trend studies have also been conducted, notably in guillemot eggs from Sweden (1969-2000), beluga whales from the Canadian Arctic (1982-1997 and 1989-2001) and from the St. Lawrence Estuary (1988-1999), and ringed seals from the Canadian Arctic (1981-2000). In the temperate latitudes, from these and other studies (e.g. in dated sediment cores), PBDE concentrations began to rise earlier than in those from high latitudes, in line with data for production and use. These trends have now slowed in many cases. Declines could be expected in Europe for many congeners following the cessation of manufacture and use of the penta-mix formulation in the EU, though these are not yet apparent in environmental samples. In Arctic biota, however, the rapidly rising concentrations seen currently in Canada could be expected to continue for some time, reflecting continued production and use of the penta-mix formulation in North America (>95% of the world total) and the impact of long-range atmospheric transport. PMID- 12850095 TI - Polybrominated diphenyl ether flame retardants in the North American environment. AB - North America consumes over half of the world's production of polybrominated diphenyl ether (PBDE) flame retardants. About 98% of global demand for the Penta BDE mixture, the constituents of which are the most bioaccumulative and environmentally widespread, resides here. However, research on the environmental distribution of PBDEs in North America has lagged behind that in Northern Europe. Examination of available governmentally maintained release data suggests that Deca-BDE use in the US substantially exceeds that in Canada. Penta-BDE use probably follows a similar pattern. PBDE demand in Mexico is uncertain, but is assumed to be comparatively modest. Recent research examining air, water, sediment, sewage sludge and aquatic biota suggests that Penta-BDE constituents are present in geographically disparate locations in the US and Canada. The less brominated congeners have been observed in areas distant from their known use or production, e.g. the Arctic. PBDEs have been detected in low concentrations in North American air, water and sediment, but much higher levels in aquatic biota. Increased burdens as a function of position in the food web have been noted. PBDE concentrations in US and Canadian sewage sludges appear to be at least 10-fold greater than European levels and may be a useful barometer of release. In general, PBDE concentrations in environmental media reported in North America are comparable or exceed those observed elsewhere in the world. In contrast to Europe, environmental burdens are increasing over time here, consistent with the greater consumption of the commercial mixtures. However, data remain relatively scarce. Deca-BDE in the North American environment appears largely restricted to points of release, e.g. urban areas and those where PBDE-containing sewage sludges have been applied. This lack of redistribution is likely due to its extremely low volatility and water solubility. Penta-BDE and Deca-BDE products are used in different applications and this may also be a factor controlling their environmental release. PMID- 12850096 TI - BFR-governmental testing programme. AB - As a consequence of results from recent studies, indicating increasing concentrations of polybrominated diphenyl ethers (PBDEs) in a wide range of environmental samples, governments have begun to consider the need to restrict the production and use of this compound group. Within the EU, it has been decided to cease production and use of the pentamix PBDE formulation, and the industry has already moved to alternative compounds. In Asia, the Japanese industries restrict voluntarily the production and use of polybrominated biphenyls (PBBs), hexabromodiphenyl ether and tetrabromodiphenyl ether. In North America, no such decisions have been taken as yet, and production of the pentamix continues. Ecolabelling and ecological product declarations are also being used in order to accelerate the phase-out process of brominated flame retardant (BFR). They restrict to different degrees the use of BFR in plastic, textiles, flexible floorings and insulating materials. Many governments have also initiated studies intended to provide more information on the octamix and decamix PBDE formulations, and the replacement compounds hexabromocyclododecane (HBCD) and tetrabromobisphenol A (TBBPA), regarding their significance as environmental contaminants and to inform the need for further regulatory action. These studies are summarised below. PMID- 12850097 TI - Industry-sponsored research on the potential health and environmental effects of selected brominated flame retardants. AB - Modern fire-fighting techniques, equipment and fire-resistant building design has lead to less destruction than in the previous centuries. However, a high fuel load in either a residence or a commercial building can overwhelm even the best firefighters or building construction, and factors affecting the fuel load have changed in recent decades. The fire load in a typical home has doubled over the last 50 years, furnishings typically include those made of petrochemicals that can behave as if containing built-in accelerant, and modern energy-efficient buildings are less able to disperse heat in the event of a fire. Flame retardant chemicals (FRs) are one means used to reduce the risk of fire. FRs are typically added or incorporated chemically into a polymer to slow or hinder the ignition or growth of a fire in low-to-moderate cost commodity polymers. One type of FR contains bromine atoms as the active moiety. The FR industry, either as individual companies or as consortia, has conducted a broad range of studies on the commercial deca-, octa- and pentabromodiphenyl oxide/ether, tetrabromobisphenol A and hexabromocyclododecane products. These five products have data in excess of the OECD Screening Informational Data Set (SIDS) and the U.S. High Production Volume (HPV) program, and sufficient data for the performance of formal EU risk assessments. The objective of this paper is to present the range of data developed by industry consortia and to provide sources for the information. We hope to facilitate further research by assembling references to industry consortia-sponsored research here. PMID- 12850098 TI - Metabolism in the toxicokinetics and fate of brominated flame retardants--a review. AB - Several classes of brominated flame retardants (BFRs), namely polybrominated biphenyls (PBBs), polybrominated diphenyl ethers (PBDEs), tetrabromobisphenol A (TBBPA), hexabromocyclododecane (HBCCD), bis(2,4,6-tribromophenoxy)ethane (BTBPE), and tris(2,3-dibromopropyl)phosphate (Tris), have been identified as environmental contaminants. PBDEs, TBBPA, and HBCCD are of particular concern due to increasing environmental concentrations and their ubiquitous presence in the tissues of humans and wildlife from Europe, Japan, and North America. Regardless, the toxicokinetics, in particular metabolism, of BFRs has received little attention. The present review summarizes the current state of knowledge of BFR metabolism, which is an important factor in determining the bioaccumulation, fate, toxicokinetics, and potential toxicity of BFRs in exposed organisms. Of the minimal metabolism research done, BFRs have been shown to be susceptible to several metabolic processes including oxidative debromination, reductive debromination, oxidative CYP enzyme-mediated biotransformation, and/or Phase II conjugation (glucuronidation and sulfation).However, substantially more research on metabolism is necessary to fully assess BFR fate, uptake and elimination kinetics, metabolic pathways, inter-species differences, the influence of congener structure, and the potential health risks to exposed organisms. PMID- 12850099 TI - A review on human exposure to brominated flame retardants--particularly polybrominated diphenyl ethers. AB - Brominated flame retardants (BFRs) have been and are still heavily used as additive or reactive chemicals in polymers and textiles. Only a few of the BFRs have been assessed in human subjects with a major data set on internal exposures to polybrominated diphenyl ethers (PBDEs). Increasing PBDE levels have been observed in mothers' milk from Sweden as well as in blood from Germany and Norway. The levels are in general lower than PCB levels. However, the PBDE concentrations found in the North Americans are considerably higher compared to European subjects. The PBDEs are dominated by 2,2',4,4'-tetrabromodiphenyl ether (BDE-47). Decabromodiphenyl ether (BDE-209) is reported both in the general population and in occupationally exposed persons showing the bioavailability of this high molecular weight compound. While the lower and medium brominated diphenyl ethers are persistent, BDE-209 has a fairly short half-life of approximately 2 weeks. Tetrabromobisphenol A (TBBPA) is readily eliminated in humans showing a half-life of about 2 days. Still, TBBPA is accumulated in humans but a continuous exposure to this BFR is required to maintain a certain level in the human subject. TBBPA has not been detected in the general population but in people exposed at work. The current review addresses human exposure routes and levels of BFRs. PMID- 12850100 TI - Toxic effects of brominated flame retardants in man and in wildlife. AB - Brominated flame retardants (BFRs) are ubiquitous industrial chemicals, and many of them are produced in large volumes. Due to this fact, several BFRs are found in quantifiable levels in wildlife, as well as in humans. However, we are still lacking information on the effects of BFR in wildlife and, especially, in man. This review summarises the biological effects of polybrominated diphenyl ethers (PBDEs), tetrabromobisphenol A (TBBPA) and derivates, hexabromocyclododecane (HBCD) and polybrominated biphenyls (PBBs), however excluding other aspects such as environmental levels. These BFR groups were selected because of a large volume production (PBDEs, TBBPA and derivates), and availability of some toxicity data in spite of much lower production volumes (HBCD and PBBs). In addition, the increase in levels of PBDEs in human (breast milk) and wildlife samples during later time made it especially interesting to include this BFR group. PBDES: The commercial PBDE products predominantly consist of so-called penta-, octa- and decabromodiphenyl ether products. Each product consists of a rather narrow range of congeners and is named after the dominating congener as regards the bromination pattern. Generally, the PentaBDEs seem to cause adverse effects at the comparably lowest dose, whereas much higher doses were needed for effects of the DecaBDEs. The critical effects of PentaBDEs are those on neurobehavioural development (from 0.6 mg/kg body weight) and, at somewhat higher dose, thyroid hormone levels in rats and mice, of OctaBDEs on fetal toxicity/teratogenicity in rats and rabbits (from 2 mg/kg body weight), and of DecaBDEs on thyroid, liver and kidney morphology in adult animals (from 80 mg/kg body weight). Carcinogenicity studies, only performed for DecaBDEs, show some effects at very high levels, and IARC (1990) evaluates DecaBDEs not classifiable as to its carcinogenicity to humans. TBBPA: The toxicity of TBBPA in the experimental in vivo studies is suggested to be low. In most reported studies, only doses in g/kg body weight were effective, but at least one study suggested renal effects at around 250 mg/kg body weight. Although difficult to include and interpret in a quantitative risk assessment, the in vitro effects on immunological and thyroid hormones, as well as binding to erythrocytes should be noted. Before a solid standpoint could be reached on TBBPA toxicity additional studies must be performed. This statement is even more valid regarding the TBBPA derivates, where there is an almost complete lack of toxicity data. HBCD: Also in the case of HBCD, relevant toxicity studies are lacking. Based on the present animal studies, a critical effect is seen in the liver and on thyroid hormones (LOAEL 100 mg/kg body weight/day). However, in a recent short paper behavioural effects in mice pups were observed already at 0.9 mg/kg body weight, and behavioural effects may be a sensitive endpoint for HBCD, as well as for other BFRs. PBBS: Due to the Michigan accident in 1973-1974, many toxicity studies on PBBs are available. The critical experimental effects are those on reproduction and carcinogenicity, and a NOAEL of 0.15 mg/kg body weight/day could be suggested based on the cancer effects. In man no unequivocal effects have been observed, although in some studies neurological and musculoskeletal symptoms were suggested. Based on the carcinogenic effects in animals, a human TDI of 0.15 microg/kg body weight has been presented. To conclude, the toxicity data are almost entirely based on experimental models. There are differences among the BFR groups, as well as within these groups, both regarding type of toxic effect and at what dose it appears. As BFRs will continue to appear both in industrial applications and, even if the production has ceased, in our environment, there is a continued need for effects studies on BFRs. PMID- 12850101 TI - Health effects of polybrominated dibenzo-p-dioxins (PBDDs) and dibenzofurans (PBDFs). AB - This article reviews the state of the science regarding the health effects of polybrominated dibenzo-p-dioxins (PBDDs) and dibenzofurans (PBDFs). While thousands of articles have been published on the health effects of 2,3,7,8 tetrachlorodibenzo-p-dioxin and related polychlorinated dibenzodioxins (PCDDs) and dibenzofurans (PCDFs), little is know about the brominated and mixed chloro/bromo homologs. Available literature suggests that brominated compounds have similar toxicity profiles to their chlorinated homologs. However, further research investigating health effects will only be useful if exposure scenarios truly exist. Current exposure data is extremely limited, posing a major data gap in assessing potential risk of these chemicals. The rapid increase in the use of brominated flame retardants has raised the level of environmental concern regarding PBDDs/PBDFs as it is likely that human, as well as wildlife, exposure to brominated dioxins and furans will increase with their use. PMID- 12850102 TI - Brominated dioxin-like compounds: in vitro assessment in comparison to classical dioxin-like compounds and other polyaromatic compounds. AB - Recently, several countries agreed to adopt the Stockholm convention on persistent organic pollutants (POPs). One future obligation will be to add other POPs as new evidence becomes available. In vitro cell-based bioassays offer a rapid, sensitive, and relatively inexpensive solution to screen possible POP candidates. In the present study, we investigated the aryl hydrocarbon (Ah) receptor activity of several dioxin-like POPs by using the Micro-EROD (Ethoxy Resorufin-O-Deethylase) and DR-CALUX (Dioxin-Responsive-Chemical Activated Luciferase gene eXpression) bioassays, which are two state-of-the-art methods. The Micro-EROD system used in our study utilizes a wild-type rat liver cell line (rat liver H4IIEC3/T cells), while the DR-CALUX bioassay consists of a genetically modified rat hepatoma H4IIE cell line that incorporates the firefly luciferase gene coupled to dioxin-responsive elements (DREs) as a reporter gene. In the case of the DR-CALUX bioassay, we used an exposure time of 24 h, whereas we used a 72-h exposure time in the Micro-EROD bioassay. The aim of this study was to compare conventional dioxin-like POPs (such as polychlorinated dibenzodioxins and -furans, PCDD/Fs and coplanar polychlorinated biphenyls, PCBs) with several other classes of possible candidates to be added to the current toxicity equivalent factor (TEF) model in the future. Therefore, this study compares in vitro CYP1A1 (Micro-EROD bioassay) and firefly luciferase induction (DR-CALUX bioassay) in several mixed polyhalogenated dibenzodioxins and -furans (PXDD/Fs; X=Br, Cl, or F), alkyl-substituted polyhalogenated dibenzodioxins and furans (PMCDD/Fs; M=methyl), polyhalogenated biphenyls (PXBs, X=Br, Cl ), polybrominated diphenyl ethers (PBDEs), pentabromophenols (PBPs), and tetrabromobisphenol-A (TBBP-A). We also evaluate congener-specific relative potencies (REPs) and efficacies (% of TCDD(max)) and discuss the dose-response curves of these compounds, as well as the dioxin-like potency of several other Ah receptor agonists, such as those of the polyaromatic hydrocarbons (PAHs) and polychlorinated naphthalenes (PCNs). The highest REP values were found for several PXDD/F congeners, followed by some coplanar PXBs, trichlorinated PCDD/Fs, PAHs, PBDE-126, 1-6-HxCN, and some brominated flame retardants (TBBP-A). These in vitro investigations indicate that further research is necessary to evaluate more Ah-receptor agonists for dioxin-like potency. PMID- 12850103 TI - Are brominated flame retardants endocrine disruptors? AB - Brominated flame retardants (BFRs) are a group of compounds that have received much attention recently due to their similarity with "old" classes of organohalogenated compounds such as polychlorinated biphenyls (PCBs), in terms of their fate, stability in the environment and accumulation in humans and wildlife. Toxic effects, including teratogenicity, carcinogenicity and neurotoxicity, have been observed for some BFR congeners, in particular the brominated diphenyl ethers (BDEs). This concise review focuses on the potency of BFRs and to disrupt endocrine systems, and attempts to answer the question whether or not BFRs are endocrine disruptors. Evidence is provided on the disruption of the thyroid hormone system by BFRs, with particular emphasis on the BDEs, as most recent data is available on this class of flame retardants. Similar to the hydroxylated PCBs, in vitro mechanistic studies as well as animal experiments have demonstrated the effects of BDEs on thyroid hormone transport and metabolism. An overview of possible effects of BFRs on the estrogen system is also provided. Research gaps are outlined, as well as ongoing and future studies in the European community aimed at contributing to comprehensive risk assessments based on the endocrine disrupting effects of BFRs. PMID- 12850104 TI - Psychological distress among Thai migrant workers in Israel. AB - The purpose of this cross-sectional study was to examine the associations between migration stressors and psychological distress among Thai migrant agricultural workers in Israel, and to examine the direct and indirect contribution of socio cultural variables to this relationship. Two hundred and twenty-one Thai male workers were interviewed using a structured questionnaire that included demographic variables and occupational exposures to organophosphate pesticides (as control variables), migration stressors, intervening variables (traditional health beliefs, social support, drinking behavior, and utilization of medical services), and a psychological distress scale. In multivariate analysis, migration stressors, the migrants' traditional health beliefs, quality of current social relationships, drinking behavior, as well as age and occupational exposure were significantly associated with psychological distress. Workers who reported higher migration stressors (perceived the migration to be difficult, and often felt homesick), those with higher levels of traditional health beliefs, those whose social relationships with other Thai co-workers were poor, those who consumed either no alcohol or large amounts of alcohol, and those who reported 'problem drinking' had significantly higher levels of psychological distress. A moderating effect of the quality of social relationships with co-workers on the association between homesickness and psychological distress was found. Additionally, migrants aged 28-34 and those who were experiencing eye irritation from chemicals at work had significantly increased levels of distress. The findings demonstrate the focal role of specific migration stressors and the current socio-cultural context on psychological distress of migrant workers. PMID- 12850105 TI - Dimensions of trauma associated with posttraumatic stress disorder (PTSD) caseness, severity and functional impairment: a study of Bosnian refugees resettled in Australia. AB - Refugee survivors of inter-ethnic warfare vary greatly in the extent and range of their trauma experiences. Discerning which experiences are most salient to generating and perpetuating disorders such as posttraumatic stress disorder (PTSD) is critical to the mounting rational strategies for targeted psychosocial interventions. In a sample of Bosnian Muslim refugees (n=126) drawn from a community centre and supplemented by a snowball sampling method, PTSD status and associated disability were measured using the clinician-administered PTSD Scale (CAPS) for DSM-IV. A principal components analysis (PCA) based on a pool of trauma items yielded four coherent trauma dimensions: Human Rights Violations, Threat to Life, Traumatic Loss and Dispossession and Eviction. A cluster analysis identified three subgroupings according to extent of trauma exposure. There were no differences in PTSD risk for the group most exposed to human rights violations (internment in concentration camps, torture) compared to the general war-exposed group. Logistic regression analysis using the dimensions derived from the PCA indicated that Threat to Life alone of the four trauma factors predicted PTSD status, a finding that supports the DSM-IV definition of a trauma. Both Threat to Life and Traumatic Loss contributed to symptom severity and disability associated with PTSD. It may be that human rights violations pose a more general threat to the survivor's future psychosocial adaptation in areas of functioning that extend beyond the confines of PTSD. PMID- 12850106 TI - Factors affecting decisions to seek treatment for sick children in Kerala, India. AB - The purpose of this study was to measure the effects of social and economic variables, disease-related variables, and child gender on the decisions of parents in Kerala, India, to seek care for their children and on their choice of providers in the allopathic vs. the alternative system. A case-control analysis was done using data from the Kerala section of the 1996 Indian National Family Health Survey, a cross-sectional survey of a probability sample of households conducted by trained interviewers with a close-ended questionnaire. Of the 469 children who were eligible for this study because they had at least one common symptom suggestive of acute respiratory illness or diarrhea during the 2 weeks before the interview, 78 (17%) did not receive medical care, while the remaining 391 (83%) received medical care. Of the 391 children who received medical care, 342 (88%) received allopathic medical care, and 48 (12%) received alternative medical care. In multivariable analyses, parents chose not to seek medical care for their children significantly more often when the illness was mild, the child had a specific diagnosis, the mother had previously made fewer antenatal visits, and the family had a higher economic status. When parents sought medical care for their children, care was sought significantly more often in the alternative provider system when the child was a boy, the family lived in a rural area, and the family had a lower social class. We conclude that, in Kerala, disease severity and economic status predict whether children with acute respiratory infection or diarrhea are taken to medical providers. In contrast, most studies of this issue carried out in other populations have identified economic status as the primary predictor of medical system utilization. Also in Kerala, the gender of the child did not influence whether or not the child was taken for treatment but did influence whether care was sought in the alternative or the allopathic system. PMID- 12850107 TI - Interacting with cancer patients: the significance of physicians' communication behavior. AB - A diagnosis of cancer typically results in patients experiencing uncertainty about and loss of control over their situation, which in turn has a negative influence on their health outcomes. Cancer treatment further disrupts patients' quality of life. Throughout their cancer journey patients often rely on their physicians to provide them with social/interpersonal, informational, and decisional support. A growing body of research shows that physicians' communication behavior does indeed have a positive impact on patient health outcomes. Thus, the patient-physician interaction assumes great significance in the cancer care delivery process. It is encouraging to note that research in this area, largely dominated by studies conducted in primary care, is attracting the attention of cancer researchers. In an attempt to encourage and aid future research on patient-physician communication in cancer care, this paper presents a critical evaluation of existing literature on key elements of physicians' communication behavior (i.e., interpersonal communication, information exchange, and facilitation of patient involvement in decision-making). Different approaches to assessing physician behavior are discussed followed by a review of key findings linking physician behavior with cancer patient health outcomes. Finally, potential limitations of existing research are highlighted and areas for future research are identified. PMID- 12850108 TI - Job characteristics, physical and psychological symptoms, and social support as antecedents of sickness absence among men and women in the private industrial sector. AB - Most longitudinal studies on the relationship between psychosocial health resources and risks, and the employees' subsequent sickness absences have been conducted in the public sector. The purpose of this study was to find out psychosocial antecedents of sickness absenteeism in the private industrial sector. The effects of job characteristics (job autonomy and job complexity), physical and psychological symptoms, and social support (from coworkers and supervisors) on sickness absenteeism were investigated. The number of long (4-21 days) and very long (>21 days) sickness absence episodes of 3895 persons (76% men and 24% women, mean age 44 years) was obtained from the health registers of a multinational forest industry corporation in 1995-1998. A questionnaire survey on the working conditions and health of the workers was carried out in 1996. The follow-up time of the sickness absences was 1-year 9-month. Job autonomy was found to be associated with long and very long episodes in men (rate ratio (RR) in the lowest autonomy group approximately 2 times higher than the highest autonomy group), and with very long episodes of absence in women (2-3 times higher RR between the low vs. the high category). Low job complexity predicted men's very long absences (RR 1.4). Long and very long episodes were associated with physical and psychological symptoms (RR 1.2-1.7) among men and women. Lack of coworkers' support increased the frequency of very long sickness absence among men (RR 1.4), and lack of supervisor's support among women (RR 1.6). Also, some interaction effects of social support variables were observed among both genders. We conclude that the studied psychosocial factors are associated with subsequent sickness absence, and that the associations are partly gender-specific. The results showing which variables are related to employees' sickness absenteeism in the private industrial sector can be applied in human resource management and health service planning. PMID- 12850109 TI - Children's health-related quality of life, neighbourhood socio-economic deprivation and social capital. A contextual analysis. AB - Neighbourhood objective socio-economic indicators and community-reported subjective measures of social capital were examined in relation to children's health-related quality of life in the Netherlands. Three different data-sources were used: (1) objective neighbourhood socio-economic indicators, (2) subjective neighbourhood data on social capital, and (3) individual data of a family cohort study, including questions on children's health-related quality of life, and family socio-economic status. Multilevel analyses were conducted using both neighbourhood level and individual level data. Neighbourhood socio-economic status and social capital were associated. Measures of socio-economic deprivation and social capital were both non-specifically associated with children's general health and satisfaction, independent of possible individual-level confounders. However, children's mental health and behaviour were specifically associated with one aspect of social capital, the degree of informal social control in the neighbourhood. PMID- 12850110 TI - Poverty, affluence, and income inequality: neighborhood economic structure and its implications for health. AB - In this paper, we attempt to verify that neighborhood economic structure influences individual health over and above other individual characteristics, and to comparatively evaluate the effects of neighborhood concentrated affluence, concentrated poverty and income inequality in relation to individual health in the USA. We also explore physical environment, health-enhancing services, social hazards (crime) and social resources as mechanisms operating at the neighborhood level that may help to explain the influence of structural economic conditions on health. We use Hierarchical Ordinal Logit Models to examine a rich multi-level data set. Results indicate that affluence exerts significant contextual effects on self-rated health while poverty and income inequality at the neighborhood level are not important structural factors. Moreover, we find that a composite measure of social resources distinguishes itself in both explaining the impact of concentrated affluence and exerting an independent contextual effect on individual health. Physical environment, or the level of physical disorder in the neighborhood, also mediates the effect of affluence on self-rated health, although to a lesser degree than social resources. Our empirical findings suggest that different dimensions of economic structure do not in fact have unique and additive contributions to individual health; the presence of affluent residents is essential to sustain neighborhood social organization which in turn positively affect health. PMID- 12850111 TI - Socioeconomic inequalities in morbidity among the elderly; a European overview. AB - There is some evidence on socioeconomic inequality in morbidity among elderly people, but this evidence remains fragmentary. This study aims to give a comprehensive overview of educational and income inequalities in morbidity among the elderly of eleven European countries. Data from the first wave of 1994 of the European Community Household Panel were used. The study population comprised a total of 14,107 men and 17,243 women, divided into three age groups: 60-69, 70-79 and 80+. Three health indicators were used: self-assessed health, cut down in daily activities due to a physical or mental problem, and long-term disability. The results indicate that socioeconomic inequalities in morbidity by education and income exist among the elderly in Europe, in all the countries in this study and all age groups, including the oldest old. Inequalities decline with age among women, but not always among men. Greece, Ireland, Italy and The Netherlands most often show large inequalities among men, and Greece, Ireland and Spain do so among women. To conclude, inequalities in morbidity decrease with age, but a substantive part persists in old age. To improve the health of elderly people it is important that the material, social and cultural resources of the elderly are improved. PMID- 12850112 TI - Quality of internet access: barrier behind internet use statistics. AB - The rapid growth of the Internet is increasingly international with young people being the early adopters in most countries. However, the quality of Internet access looms as a major barrier hidden behind Internet use statistics. The goal of this study was to provide an in-depth evaluation of young people's perspectives on using the Internet to obtain health information and resources (e health). Using an inductive qualitative research design, 27 focus groups were conducted in Ontario, Canada. The 210 young participants were selected to reflect diversity in age, sex, geographic location, cultural identity and risk. A major finding was how the quality of Internet access influenced young people's ability to obtain health information and resources. Quality of Internet access was affected by four key factors: 1. Privacy, 2. Gate-keeping, 3. Timeliness and 4. Functionality. Privacy was particularly relevant to these young people in getting access to sensitive health information (e.g. sexual activities). Variations in access quality also impacted participation in mutual support, fostering social networks and getting specific health questions answered. These results serve as a warning about using Internet penetration statistics alone as a measure of access. Concerted attention is needed on improving the quality of Internet access for achieving the potential of e-health. This is imperative for addressing the digital divide affecting populations both within countries and globally between countries. PMID- 12850113 TI - Damned if you do: culture, identity, privilege, and teenage childbearing in the United States. AB - Why is the broad American public disapproving of urban African American teen mothers and unaware that the scientific evidence on the consequences of teen childbearing, per se, is equivocal? I focus on the links between culture, identity, and privilege. I argue that the broader society is selective in its attention to the actual life chances of urban African Americans and how these chances shape fertility-timing norms, in part, because this selective focus helps maintain the core values, competencies, and privileges of the dominant group. Delayed childbearing is an adaptive practice for European Americans and an intensely salient goal they have for their children. Yet early fertility-timing patterns may constitute adaptive practice for African American residents of high poverty urban areas, in no small measure because they contend with structural constraints that shorten healthy life expectancy. European Americans put their cultural priorities into action ahead of the needs of African Americans and employ substantial resources to disseminate the social control message meant for their youth that teenage childbearing has disastrous consequences. Their ability to develop a more nuanced understanding of early childbearing is limited by their culturally mediated perceptions. Thus, cultural dominance can be perpetuated by well-meaning people consciously dedicated to children's well-being, social justice, and the public good. The entrenched cultural interdependence of and social inequality between European and African Americans leads African Americans to be highly visible targets of moral condemnation for their fertility behavior, and also sets up African Americans to pay a particularly high political, economic, psychosocial, and health price. PMID- 12850114 TI - Ethnicity, socio-economic position and gender--do they affect reported health care seeking behaviour? AB - While the pursuit of equity of access to health care is a central objective of many health care systems, there is evidence that patients of ethnic minority descent, in lower socio-economic position (SEP) or of female gender are less likely than Whites, more affluent groups or men, respectively, to access secondary and tertiary medical care. However, it is unclear at which point in the chain of events leading from perception of need through attendance at primary/emergency care, to referral and receipt of secondary care, this inequality occurs. This study examined the influence of ethnicity, socio-economic position and gender on an individual's perception of the need and urgency for seeking health care. A random sample was selected from two large city General Practices in the UK who were sent postal questionnaires which included two clinical vignettes describing characters experiencing chest pain and discovering a lump in the armpit. The main outcome measure was response to the 'chest pain' and 'lump' vignettes in terms of immediate health care utilisation. The questionnaire survey (n=1350, response rate 66%) indicated that Black respondents, respondents from lower socio-economic groups and women were at least as likely to report immediate health care seeking in response to the clinical vignettes than White respondents, those from higher socio-economic groups or men. This finding was consistent across all scenarios after adjustment for interpretation of the vignette, access to health services and attitudes to health and health care. For example, those in the lowest SEP group were almost 60% more likely to report immediate care seeking in response to the lump vignette (OR 1.59, 95% CI 1.08-2.33) compared to those in the highest SEP group; and Black respondents 40% more likely (OR 1.41, 95% CI 0.92-2.17). This study suggests inequalities in access to health care by ethnicity, socio-economic position and gender are not related to patients in these groups failing to self-refer to primary or accident and emergency care, barriers must therefore occur at the level of health care provision. PMID- 12850115 TI - Narrative strategies in medical discourse: constructing the psychiatric 'case' in a non-western setting. AB - The organizing goal of the present study was to analyze and understand the 'discursive presentation' of the Egyptian psychiatric patient through the texts, or narratives, contained within the patient medical charts. It is argued that the medical record, as a written document blending overt medical imperatives with more unexamined cultural assumptions about self-hood and abnormality, is an unusually rich source of discursive data concerning the 'cultural negotiations' implicit in the construction of the patient according to the two (often competing) world views represented by western biomedicine and traditional Egyptian culture. Psychiatry in Egypt is much more than a 'foreign transplant'; to assume this is to deny the culturally constructed nature of western biomedicine and psychiatry, which have their roots in historical and cultural notions of self, society, the individual, and normality versus abnormality (Transcultural Psychiat. 35(3) (1998) 352). Egyptian psychiatry is the product of an ongoing active blending of two very different conceptualizations of these issues. PMID- 12850116 TI - Discourses of blame: accounting for aggression and violence on an acute mental health inpatient unit. AB - The English National Service Framework for Mental Health stipulates that the highest quality of health care should be provided for mental health service users in the most efficient and effective manner. Incidents of aggression and violence militate against achieving that goal, yet such incidents are frequently reported in inpatient settings. Traditionally, research in this area has focused on the extent of the phenomenon, the individual characteristics of those involved and precursors to the incident. For the most part the literature reflects a dualistic, perpetrator/victim conceptualisation of incidents. This study aimed to address the lack of research undertaken from a more systemic perspective by examining how all those involved understood and attributed meaning to violent or aggressive situations and how these attributions justified individual perceptions, reactions and actions. Working from the position that all behaviour, including violent behaviour, has meaning to those involved and can be understood, 16 semi-structured interviews were carried out in one mental health unit. Because only one client was both willing and able to give a full account of an incident, we focus here on two incidents in which that client was involved. Discourse analytic techniques were used to examine her account of the two incidents and those of the staff members involved. Participants discussed key themes from the interviews in terms of several dilemmas: whether the violent or aggressive behaviour was 'mad' or 'bad'; predictable or unpredictable; and had resulted from 'personality' or ' mental illness'. The client and staff discourses were strikingly similar and in each case the central concern was with the attribution of blame. The findings have implications for the professional discourse of mental health care, including the discourse of the current policy agenda, a discourse itself constructed with the primary function of exoneration from and attribution of blame. PMID- 12850117 TI - Patients and doctors: reformulating the UK health policy community? AB - The rise of the active health care consumer in the United Kingdom requires a reformulation not only of the traditional relationship between patients and doctors, but also of the macro-politics of health which reflect and service that relationship. Market and democratic themes have supplied an ideological impetus to the pressures for change. The well-publicised problems of medical self regulation have given them practical political expression. However, the response from the policy community still reflects the dominant partners within it, medicine and the state. What neither partner has recognised is that the functionality of the policy community has been undermined by the different and issue-based challenges to the traditional patient-doctor relationship. As a result, the state is likely to remain the lead player in an increasingly unstable politics of health where consumerist issues are on the policy agenda, but patient groups are still excluded from the policy community. PMID- 12850118 TI - Public and private sector responses to essential drugs policies: a multilevel analysis of drug prescription and selling practices in Mali. AB - Many African countries have introduced cost recovery mechanisms based on the sale of drugs and measures aimed at improving drug supply. This study compares prescribing and selling practices in Mali, in 3 cities where the public sector contributes differentially to the supply of drugs on the market. Multilevel models are used to analyse the content and cost of 700 medication transactions observed in 14 private and public legal points of sale. Results show that the objective of improving access to drugs seems to have been achieved in the sites studied. Costs of prescriptions were lower where public health services had been revitalized. Affordable generic drugs were accessible and widely used, even in the private sector. However, measures intended to rationalize the prescription and delivery of drugs did not always have the desired effect. While agents in the public sector tended to prescribe fewer antibiotics, injectables, or brand-name drugs, the data confirm the virtual absence of advice concerning the use or the side effects of the drugs in both public and private sectors. In addition, data supported the notion that the public and private sectors are closely intertwined. Notably, availability of drugs in the public sector contributed to diminishing the prices charged in the private sector. Similarly, the use that agents in the public sector made of the opportunities afforded by the presence of the private pharmaceutical sector provided another illustration of interrelatedness. Finally, the data showed that the presence of a private sector, which has not been affected by measures aimed at rationalizing prescription and sales practices, limits the effects of measures implemented in the public sector. More assertive policies, based on strategies encompassing actors in the private sector, are needed to increase the safety and effectiveness of prescription and sales practices. PMID- 12850119 TI - The DALY, context and the determinants of the severity of disease: an exploratory comparison of paraplegia in Australia and Cameroon. AB - This paper summarises the findings of an empirical investigation of some of the technical and social assumptions on which the disability adjusted life year (DALY) is based. The objectives of the study were to examine the notion that the burden of disease is broadly similar without regard to country, environment, gender or socio-economic status and to develop detailed descriptions of the experiences of the burden of disease as they related to these contextual factors. The study was a multi-factorial exploratory study employing qualitative and quantitative techniques to obtain data on the effects of country (development), environment (urban versus rural), gender and socio-economic status on people with paraplegia. The data provided an extensive and detailed compilation of context rich descriptions of living with paraplegia. Striking features of the data were the differences between countries with respect to the impact of the health conditions on functioning and highlight a context in which paraplegia of like clinical severity can be fatal in one environment and not in another. While there has been some focus on the control of social determinants of disease, there has been little work on the social determinants of the severity of disease. The underlying assumptions of the DALY, which ignore context in the assessment of the burden of disease, risk exacerbating inequalities by undervaluing the burden of disease in less-developed countries. There is a need to continue to subject the development of indicators to rigorous debate to determine a balance between the assumption of a global "average social milieu" and the treatment of each individual as belonging to their own context in the assessment of population health in order for indicators to be meaningful cross-culturally. PMID- 12850120 TI - Transforming growth factor-beta and its role in asthma. AB - Transforming growth factor-beta (TGF-beta) is an important fibrogenic and immunomodulatory factor that may play a role in the structural changes observed in the asthmatic airways. In vitro as well as in vivo studies have evidenced a dual role for TGF-beta: it can either function as a pro- or anti-inflammatory cytokine on inflammatory cells, participating into the initiation and resultion of inflammatory and immune responses in the airways. TGF-beta is also involved in the remodelling of the airway wall, and has in particular been related to the subepithelial fibrosis. TGF-beta is produced in the airways by inflammatory cells infiltrated in the bronchial mucosa, as well as by structural cells of the airway wall including fibroblasts, epithelial, endothelial and smooth muscle cells. By releasing TGF-beta, these different cell types may then participate into the increased levels of TGF-beta observed in bronchoalveolar lavage fluid from asthmatic patients. Taken together, these results suggest that TGF-beta may play a role in inflammation in asthma. However, as its role is dual in the modulation of inflammation, further studies are needed to elucidate the precise role of TGF beta in the airways. PMID- 12850121 TI - Long-acting beta(2) agonists as potential option in the treatment of acute exacerbations of COPD. PMID- 12850122 TI - Cough sensitivity in atopic dermatitis. AB - The aim of study was to investigate the cough sensitivity (C2) to capsaicin (CAPS) in patients with atopic dermatitis without clinical respiratory symptoms. Cough sensitivity (C2) is defined as the lowest CAPS concentration, which evokes two or more coughs. Forty eight dermatological patients (21 M, 27 F; mean age 44 yr) and 24 healthy volunteers (14 M, 10 F; mean age 37 yr) inhaled deep breath (2 l) of CAPS aerosol in doubled concentrations (from 0.02 to 200 micromol/l) (Pari Provokationstest I, PARI WERK; mass median diameter 1.2 microm). Cough sensitivity (C2) expressed as geometric mean (95% CI) of CAPS concentration was 0.13 micromol/l (0.06-0.31) in 26 patients with atopic dermatitis (10 M, 16 F; mean age 41 yr), 5.51 micromol/l (1.33-22.90) in 22 patients with psoriasis (11 M, 11 F; mean age 46 yr) and 4.29 micromol/l (2.54-7.26) in 24 controls. There is significant difference of cough sensitivity (C2) between patients with atopic dermatitis and healthy volunteers (p<0.001) and also between patients with atopic dermatitis and psoriasis (p<0.001). Cough sensitivity (C2) in atopic dermatitis patients without clinical respiratory symptoms is significantly increased. In patients with psoriasis cough sensitivity (C2) is not significantly changed. PMID- 12850123 TI - Inhibition of experimental acute pulmonary inflammation by pirfenidone. AB - Pirfenidone, a putative tumor necrosis factor-alpha (TNF-alpha) inhibitor, has recently gained recognition for its therapeutic use in the treatment of idiopathic pulmonary fibrosis. As pulmonary fibrosis may be the result of lung inflammatory processes, we examined the anti-inflammatory potential of pirfenidone in several models of acute pulmonary inflammation. In antigen-induced allergic paradigms, 24 h after antigen challenge, sensitized mice or guinea pigs develop a prominent pulmonary inflammation, reflected by a significant increase in the number of recoverable bronchoalveolar lavage (BAL) total cells and eosinophils. In both species, the pretreatment of animals with pirfenidone (10 and 30 mg/kg) resulted in a dose-dependent inhibition of the antigen-induced pulmonary inflammation, which was reflected by a significant decrease in the BAL eosinophils and total cells by the 30 mg/kg dose. In a non-allergic model of pulmonary inflammation, rats challenged with intratracheal LPS develop a significant increase in BAL neutrophils and total cells, along with significant increases in TNF-alpha and IL-6. Pretreatment with pirfenidone (3 and 30 mg/kg) showed a dose-dependent inhibition of the LPS-induced pulmonary inflammation, reflected by a significant decrease in the number of BAL total and neutrophilic cells at both the 3 and 30 mg/kg dose. However, pirfenidone had no effect on the peak BAL levels of TNF-alpha. In contrast, pirfenidone significantly inhibited BAL levels of IL-6. In summary, we have shown that pirfenidone can inhibit allergic and non-allergic inflammatory cell recruitment and that its pulmonary anti-inflammatory activity is independent of TNF-alpha inhibition. PMID- 12850124 TI - Methylprednisolone effects on oxygenation and histology in a rat model of acute lung injury. AB - We examined the effects of methylprednisolone on gas exchange, pressure-volume curve, lavage fluid inflammatory cell counts, protein content, surfactant pool size, and lung histology in a rat model of paraquat-induced lung injury. Twenty three adult male Sprague-Dawley rats received intraperitoneal paraquat injection (35 mg/kg) and were randomly divided into three groups: (1). control group received no further treatment; (2). 1-dose methylprednisolone group received a concomitant intraperitoneal methylprednisolone injection (30 mg/kg); (3). 3-dose methylprednisolone group received a concomitant and daily intraperitoneal methylprednisolone injection (30 mg/kg) for three doses. Three days after paraquat injection, the rat was ventilated for 90 min, a static pressure-volume curve and bronchoalveolar lavage was performed, and postmortem histology was examined. Surfactant pool size of the 3-dose methylprednisolone group was significantly increased when compared with the control and 1-dose methylprednisolone groups. Methylprednisolone treatment increased oxygenation and the value was statistically significant for 3-dose methylprednisolone group at 90 min of ventilation. Inflammatory cell counts in bronchoalveolar lavage fluid and lung injury score were decreased as the methylprednisolone dose increased. We conclude that high-dose methylprednisolone treatment increased surfactant pool size and improved lung histology of paraquat-injured lungs but this augmentation could not significantly improve oxygenation throughout the ventilation period. PMID- 12850125 TI - Elastase-induced changes in lung function: relationship to morphometry and effect of drugs. AB - Intratracheal administration of porcine pancreatic elastase (PPE) produced a dose related decline in lung function, as assessed by changes in dynamic lung compliance (C(dyn)) in New Zealand White rabbits. This occurred within 24 h of administration and persisted for 56 days (n=6). These lung function changes were accompanied by histological evidence of emphysema in the lungs and were not mimicked by intratracheal administration of the proteolytic enzyme trypsin. Neither the lung function nor the histological changes induced by elastase could be prevented or reversed by either the glucocorticosteroid, dexamethasone, or all trans retinoic acid (ATRA).Our data suggest that local administration of elastase to the lungs of rabbits may provide a convenient way to assess the effects of drugs on the changes induced by elastase in airways. PMID- 12850126 TI - Levodropropizine does not affect P0.1 and breathing pattern in healthy volunteers and patients with chronic respiratory impairment. AB - To evaluate whether the peripherally acting antitussive levodropropizine could affect the respiratory drive and the breathing pattern, we performed a double blind, randomised, cross-over trial in 12 healthy volunteers and 12 patients with chronic respiratory impairment associated with chronic obstructive pulmonary disease. Levodropropizine 6% drops (at the recommended dose for adults) or placebo were administered orally t.i.d. for 10 consecutive administrations. Mouth occlusion pressure (P0.1), minute ventilation (V(e)), tidal volume (V(t)), respiratory rate (RR), mean inspiratory flow (V(t)/T(i)), end-tidal CO(2) (EtCO(2)), oxygen saturation (SaO(2)), forced expiratory volume in 1 s (FEV(1)), and the response to a hypercapnic stimulus were measured before and 1 h after the first and the last drug administration. Levodropropizine did not modify P0.1 in basal conditions and after a hypercapnic stimulus, either in healthy volunteers or in patients. In parallel, levodropropizine did not significantly affect V(t), RR, V(e), V(t)/T(i) and EtCO(2) in both the populations. Minor changes were induced by levodropropizine on SaO(2) in healthy volunteers, which despite a statistical difference, were too low to gain a clinical significance. These results confirmed the respiratory safety of levodropropizine 6% drops administered at the recommended dosage either in healthy volunteers or patients with chronic respiratory impairment. PMID- 12850127 TI - Short-term effect of the addition of leukotriene receptor antagonists to the current therapy in severe asthmatics. AB - In a single blind study, the short-term efficacy of the addition of leukotriene receptor antagonists (LTRA: montelukast 10 mg o.d. in 15 subjects, zafirlukast 20 mg b.i.d. in 11 subjects) to the current therapy was evaluated in severe asthmatics, unstable under regular treatment with high dose inhaled corticosteroids, bronchodilators and, in seven of them, oral corticosteroids. Each subject monitored symptoms, PEF and rescue medication during two weeks with the addition of placebo, and during two following weeks with the addition of LTRA; clinic FEV1 was measured at the beginning and at the end of each 2 weeks period. There was no significant difference in the mean FEV1, PEF, symptom score and rescue medication use between two periods of placebo and LTRA treatments. When two subjects with asthma exacerbation during treatment with LTRA were excluded, FEV1 was higher after LTRA than after placebo treatment (p=0.055). An increase in FEV1>12% pred. at the end of LTRA treatment was observed in five out of 26 subjects (19%). We suggest that LTRA have no overall significant efficacy in severe asthmatics not controlled by high dose inhaled corticosteroids and bronchodilators, but that a minority of these patients could be particularly sensitive to the positive effects of these drugs. The detection of these 'responders' could be relevant in the treatment of severe asthma. PMID- 12850128 TI - Salmeterol & fluticasone 50 microg/250 microg bid in combination provides a better long-term control than salmeterol 50 microg bid alone and placebo in COPD patients already treated with theophylline. AB - Bronchodilator agents are central to the symptomatic management of Chronic Obstructive Pulmonary Disease (COPD), and long-acting inhaled bronchodilators are regarded as more convenient. The role of inhaled corticosteroids still remains controversial, but there is increasing evidence that they may improve FEV(1) and symptoms in the long-term. AIM: of the present small pilot study was to compare Salmeterol & Fluticasone (SM&FP) 50/250 microg bid via a single Diskus inhaler with SM 50 microg bid alone, and with placebo (P) in the treatment of moderate COPD. METHODS: Eighteen moderate COPD patients (53-77 yr, mean basal FEV(1)=49.1% pred.+/-5.0 s.d.; mean FEV(1) reversibility=3.6% bsln+/-3.8 s.d.) treated with theophylline 400 mg/day and beta(2) short acting prn, were divided into three matched groups of six subjects according to a double-blind design, and treated with SM&FP 50/250 microcg, or SM 50 microcg alone, or P via Diskus inhaler bid for 52 weeks. In bsln, after 4, 12, 24, 36 and 52 weeks, FEV(1) (% pred), morning PEF (l/s), the daily symptom score, and the number of exacerbations (compared with the previous year) were considered. Statistics. t-test, anova in each treatment group, and anova among basal values and among the 52 week values were used, being p<0.05 accepted. Also changes (DeltaFEV(1)) from baseline were compared at different control times. RESULTS: The mean number of exacerbations/yr decreased from 3.5+/-0.8 to 1.16+/-0.75 s.d. exacerbation/yr in the SM&FP group (t-test p<0.001); from 3.0+/-0.89 to 2.3+/-0.81 s.d. in the SM group (t-test p=ns); and from 3.16+/-1.16 to 4.16+/-0.75 s.d. in the P group (t-test p=ns). Patients receiving SM&FP showed the highest mean improvement in FEV(1) (+7.3%+/ 3.3 s.d.) over the baseline pre-treatment value after 36 weeks of treatment (anova p<0.001), being FEV(1) unchanged after 52 weeks of treatment in SM group (+0.33%+/-2.4 s.d.) and with a substantial decrease following P (-2.6%+/-1.2 s.d.) (anova p<0.001). Morning PEF (l/min) increased in subjects treated with SM&FP (anova p<0.001), while it remained unchanged in SM and P group (in both, anova p=ns). After 52 weeks of treatment, only subjects treated with SM&FP showed a reduction of the daily symptoms score from 3.6+/-0.7 to 2.0+/-0.2 s.d. (anova p=0.008). Daily beta(2) short acting prn consumption was reduced only in SM&FP group from 4.2+/-0.81 to 2.2+/-1.2 s.d. after 52 weeks (anova p<0.001). CONCLUSIONS: SM&FP 50/250 microcg regularly assumed in combination via a single Diskus inhaler for a 52 week period improves respiratory function (such as FEV(1), morning PEF), and and symptom score significantly in moderate COPD previously treated with theophylline, and at an higher extent than SM alone or P. The use of beta(2) short acting prn is also reduced, together with the number of exacerbations. PMID- 12850129 TI - Review of the BRENDA Database. PMID- 12850130 TI - A metabolic network analysis & NMR experiment design tool with user interface driven model construction for depth-first search analysis. AB - A Windows program for metabolic engineering analysis and experimental design has been developed. A graphical user interface enables the pictorial, "on-screen" construction of a metabolic network. Once a model is composed, balance equations are automatically generated. Model construction, modification and information exchange between different users is thus considerably simplified. For a given model, the program can then be used to predict all the extreme point flux distributions that optimize an objective function while satisfying balances and constraints by using a depth-first search strategy. One can also find the minimum reaction set that satisfies different conditions. Based on the identified flux distributions or linear combinations, the user can simulate the NMR and GC/MS spectra of selected signal molecules. Alternately, spectra vectorization allows for the automated optimization of labeling experiments that are intended to distinguish between different, yet plausible flux extreme point distributions. The example provided entails predicting the flux distributions associated with deleting pyruvate kinase and designing 13C NMR experiments that can maximally discriminate between the flux distributions. PMID- 12850131 TI - Production process monitoring by serial mapping of microbial carbon flux distributions using a novel sensor reactor approach: I--Sensor reactor system. AB - A novel Sensor Reactor technology is presented which permits 13C labeling experiments for metabolic flux analysis during large-scale, semi-industrial, (fed ) batch fermentation processes deriving a series of flux maps that document fermentation courses in detail. The small-scale Sensor Reactor can be inoculated within 1.50-1.20s via a special inoculation unit with an inoculation volume accuracy of 1.025+/-0.021 L. The large-scale production reactor (here: 300 L) and the Sensor Reactor were run in parallel master/slave modes to control the current pH, temperature, pressure and dissolved oxygen values as changing set points for the Sensor Reactor. Using an automated pulsing technology, glucose pulses of 5 g/L could be realized within 0.51 s. The similarity of fermentations in the Sensor Reactor with the production process was demonstrated by studying L-lysine production with C. glutamicum during multiple, 'simulated' labeling experiments each lasting 2.5h. 'Real' labeling experiments are presented in Part II. PMID- 12850132 TI - Production process monitoring by serial mapping of microbial carbon flux distributions using a novel Sensor Reactor approach: II--(13)C-labeling-based metabolic flux analysis and L-lysine production. AB - Corynebacterium glutamicum is intensively used for the industrial large-scale (fed-) batch production of amino acids, especially glutamate and lysine. However, metabolic flux analyses based on 13C-labeling experiments of this organism have hitherto been restricted to small-scale batch conditions and carbon-limited chemostat cultures, and are therefore of questionable relevance for industrial fermentations. To lever flux analysis to the industrial level, a novel Sensor Reactor approach was developed (El Massaoudi et al., Metab. Eng., submitted), in which a 300-L production reactor and a 1-L Sensor Reactor are run in parallel master/slave modus, thus enabling 13C-based metabolic flux analysis to generate a series of flux maps that document large-scale fermentation courses in detail. We describe the successful combination of this technology with nuclear magnetic resonance (NMR) analysis, metabolite balancing methods and a mathematical description of 13C-isotope labelings resulting in a powerful tool for quantitative pathway analysis during a batch fermentation. As a first application, 13C-based metabolic flux analysis was performed on exponentially growing, lysine-producing C. glutamicum MH20-22B during three phases of a pilot scale batch fermentation. By studying the growth, (co-) substrate consumption and (by-) product formation, the similarity of the fermentations in production and Sensor Reactor was verified. Applying a generally applicable mathematical model, which included metabolite and carbon labeling balances for the analysis of proteinogenic amino acid 13C-isotopomer labeling data, the in vivo metabolic flux distribution was investigated during subsequent phases of exponential growth. It was shown for the first time that the in vivo reverse C(4)-decarboxylation flux at the anaplerotic node in C. glutamicum significantly decreased (70%) in parallel with threefold increased lysine formation during the investigated subsequent phases of exponential growth. PMID- 12850133 TI - Control analysis of mitochondrial metabolism in intact hepatocytes: effect of interleukin-1beta and interleukin-6. AB - Interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) are produced by hepatic nonparenchymal cells after systemic injury and have been reported to inhibit ATP synthesis in hepatocytes, which may contribute to hepatic dysfunction in inflammatory states. To elucidate the mechanisms of action of IL-1beta and IL-6 on hepatocellular ATP synthesis, we measured the oxygen uptake rate (OUR) and mitochondrial membrane potential (MMP) of stable hepatocyte cultures, and analyzed the dynamic MMP response following the addition of mitochondrial inhibitors (antimycin A and oligomycin) with a model of mitochondrial metabolism. IL-1beta reduced mitochondrial OUR coupled to ATP synthesis via inhibition of phosphorylation reactions which dissipate the MMP, including ATP synthesis and consumption. Furthermore, the ATP synthesis rate in cytokine-free and IL-1beta treated hepatocytes was controlled primarily by phosphorylation reactions, which corresponds to a state where the ATP synthesis rate closely follows the cellular energy demand. Thus, IL-1beta-mediated effects on electron transport and substrate oxidation reactions are not likely to significantly impact on ATP synthesis. IL-6 did not reduce mitochondrial OUR coupled to ATP synthesis, but shifted the control for ATP synthesis towards processes which generate the MMP, indicating that IL-6 induces a metabolic state where cellular functions are limited by the mitochondrial energy supply. PMID- 12850134 TI - Metabolic engineering of apoptosis in cultured animal cells: implications for the biotechnology industry. AB - Animal cells have been widely used to obtain a wide range of products for human and animal healthcare applications. However, the extreme sensitivity of these cells in respect to changes experienced in their environment is evidenced by the activation of a gene-encoded program known as apoptosis, resulting in their death and destruction. From the bioprocess angle, losses in cell viability bring lower productivities and higher risks of product degradation. Consequently, many research efforts have been devoted to the development of apoptosis protective mechanisms, including the metabolic engineering of apoptosis pathways, that has proven effective in diminishing programmed cell death in a variety of biotechnological relevant cell lines. This review is focused especially in the encouraging initial results obtained with the over-expression of cloned anti apoptosis genes, from both endogenous and viral origin interfering at mitochondrial and initiator caspases levels. PMID- 12850135 TI - Transcriptional profiling of gene expression in response to glucose in Bacillus subtilis: regulation of the central metabolic pathways. AB - Chemoheterotrophic bacteria use a few central metabolic pathways for carbon catabolism and energy production as well as for the generation of the main precursors for anabolic reactions. All sources of carbon and energy are converted to intermediates of these central pathways and then further metabolized. While the regulation of genes encoding enzymes used to introduce specific substrates into the central metabolism has already been studied to some detail, much less is known about the regulation of the central metabolic pathways. In this study, we investigated the responses of the Bacillus subtilis transcriptome to the presence of glucose and analyzed the role of the pleiotropic transcriptional regulator CcpA in these responses. We found that CcpA directly represses genes involved in the utilization of secondary carbon sources. In contrast, induction by glucose seems to be mediated by a variety of different mechanisms. In the presence of glucose, the genes encoding glycolytic enzymes are induced. Moreover, the genes responsible for the production of acetate from pyruvate with a concomitant substrate-level phosphorylation are induced by glucose. In contrast, the genes required for the complete oxidation of the sugar (Krebs cycle, respiration) are repressed if excess glucose is available for the bacteria. In the absence of glucose, the genes of the Krebs cycle as well as gluconeogenic genes are derepressed. The genes encoding enzymes of the pentose phosphate pathway are expressed both in the presence and the absence of glucose, as suggested by the central role of this pathway in generating anabolic precursors. PMID- 12850136 TI - Folding of DsbB in mixed micelles: a kinetic analysis of the stability of a bacterial membrane protein. AB - Measuring the stability of integrated membrane proteins under equilibrium conditions is hampered by the nature of the proteins' amphiphilic environment. While intrinsic fluorescence is a useful probe for structural changes in water soluble proteins, the fluorescence of membrane proteins is sensitive to changes in lipid and detergent composition. As an attempt to overcome this problem, I present a kinetic analysis of the folding of a membrane protein, disulfide bond reducing protein B (DsbB), in a mixed micelle system consisting of varying molar ratios of sodium dodecyl sulfate (SDS) and dodecyl maltoside (DM). This analysis incorporates both folding and unfolding rates, making it possible to determine both the stability of the native state and the process by which the protein folds. Refolding and unfolding occur on the second to millisecond timescale and involve only one relaxation phase, when monitored by conventional stopped-flow. The kinetic data indicate that denaturation occurs around 0.3 mole fraction of SDS, in agreement with CD analysis and acrylamide quenching data. The rate constants have been fit to a three-state folding scheme involving the SDS denatured state, the native state and an unfolding intermediate that accumulates only under unfolding conditions at high mole fractions of SDS. The stability of DsbB is around 4.4 kcal/mol in DM, and this is halved upon reduction of the two periplasmic disulfide bonds, and is sensitive to mutagenesis. With the caveat that kinetic data are always open to alternative interpretations, time-resolved studies in mixed micelles provide a useful approach to measure membrane protein stability over a wide range of concentrations of SDS and DM, as well as a framework for the future characterization of the DsbB folding mechanism. PMID- 12850137 TI - Structural plasticity and the evolution of antibody affinity and specificity. AB - The germline precursor to the ferrochelatase antibody 7G12 was found to bind the polyether jeffamine in addition to its cognate hapten N-methylmesoporphyrin. A comparison of the X-ray crystal structures of the ligand-free germline Fab and its complex with either hapten or jeffamine reveals that the germline antibody undergoes significant conformational changes upon the binding of these two structurally distinct ligands, which lead to increased antibody-ligand complementarity. The five somatic mutations introduced during affinity maturation lead to enhanced binding affinity for hapten and a loss in affinity for jeffamine. Moreover, a comparison of the crystal structures of the germline and affinity-matured antibodies reveals that somatic mutations not only fix the optimal binding site conformation for the hapten, but also introduce interactions that interfere with the binding of non-hapten molecules. The structural plasticity of this germline antibody and the structural effects of the somatic mutations that result in enhanced affinity and specificity for hapten likely represent general mechanisms used by the immune response, and perhaps primitive proteins, to evolve high affinity, selective receptors for so many distinct chemical structures. PMID- 12850138 TI - Mismatch extension during strong stop strand transfer and minimal homology requirements for replicative template switching during Moloney murine leukemia virus replication. AB - Reverse transcription requires two replicative template switches, called minus and plus strand strong stop transfer, and can include additional, recombinogenic switches. Donor and acceptor template homology facilitates both replicative and recombinogenic transfers, but homology-independent determinants may also contribute. Here, improved murine leukemia virus-based assays were established and the effects of varying extents of mismatches and complementarity between primer and acceptor template regions were assessed. Template switch accuracy was addressed by examining provirus structures, and efficiency was measured using a competitive titer assay. The results demonstrated that limited mismatch extension occurred readily during both minus and plus strand transfer. A strong bias for correct targeting to the U3/R junction and against use of alternate regions of homology was observed during minus strand transfer. Transfer to the U3/R junction was as accurate with five bases of complementarity as it was with an intact R, and as few as 3nt targeted transfer to a limited extent. In contrast, 12 base recombinogenic acceptors were utilized poorly and no accurate switch was observed when recombination acceptors retained only five bases of complementarity. These findings confirm that murine leukemia virus replicative and recombinogenic template switches differ in homology requirements, and support the notion that factors other than primer-template complementarity may contribute to strong stop acceptor template recognition. PMID- 12850139 TI - Multiple interactions within the hepatitis C virus RNA polymerase repress primer dependent RNA synthesis. AB - The hepatitis C virus (HCV) RNA-dependent RNA polymerase (RdRp) initiates RNA synthesis in vivo by a de novo mechanism. In vitro, however, the HCV RdRp can initiate de novo or extend from a primed template. A novel beta-loop near the RdRp active site was previously found to prevent the use of primed templates. We found that, in addition to the beta-loop, the C-terminal tail of the HCV RdRp and the de novo initiation GTP are required to exclude the use of primed-templates. GTP binding to the NTPi site of the HCV RdRp orchestrates the participation of other structures. The interactions of the beta-loop, C-terminal tail, and GTP provide an elegant solution to ensure de novo initiation of HCV RNA synthesis. PMID- 12850140 TI - Contribution of opening and bending dynamics to specific recognition of DNA damage. AB - Guanine-uracil (G.U) wobble base-pairs are a detrimental lesion in DNA. Previous investigations have shown that such wobble base-pairs are more prone to base opening than the normal G.C base-pairs. To investigate the sequence-dependence of base-pair opening we have performed 5ns molecular dynamics simulations on G.U wobble base-pairs in two different sequence contexts, TGT/AUA and CGC/GUG. Furthermore, we have investigated the effect of replacing the guanine base in each sequence with a fluorescent guanine analogue, 6-methylisoxanthopterin (6MI). Our results indicate that each sequence opens spontaneously towards the major groove in the course of the simulations. The TGT/AUA sequence has a greater proportion of structures in the open state than the CGC/GUG sequence. Incorporation of 6MI yields wobble base-pairs that open more readily than their guanine counterparts. In order of increasing open population, the sequences are ordered as CGC0.05 vs pre ischemia perfusion). 10 microM glibenclamide and 100 microM 5-hydroxydecanoate (a selective mitochondria KATP antagonist) respectively abolished this effect of IH; calcium overloading reappeared during ischemia (1.133 +/- 0.007 and 1.118 +/- 0.007 respectively, P<0.01) and reperfusion (1.091 +/- 0.004 and 1.095 +/- 0.012 respectivly, P<0.01). However they had no effects on simulated ischemia and reperfusion-induced calcium overloading in normoxic myocytes. 50 microM pinacidil, a KATP opener, attenuated calcium overloading during ischemia and reperfusion in normoxic myocytes, but had no effect on [Ca2+]i change in IH myocytes. These results suggested that KATP channels contributed to the cardiac protection induced by IH against ischemia/reperfusion injury; the elimination of calcium overloading during ischemia/reperfusion by IH might underlie the mechanism of protection. PMID- 12850244 TI - Dietary supplementation of fermented soybean, natto, suppresses intimal thickening and modulates the lysis of mural thrombi after endothelial injury in rat femoral artery. AB - We have previously demonstrated that natto-extracts containing nattokinase (NK) inactivates plasminogen activator inhibitor type 1 and then potentiates fibrinolytic activity. In the present study, we investigated the effects of dietary supplementation with natto-extracts on neointima formation and on thrombolysis at the site of endothelial injury. Endothelial damage in the rat femoral artery was induced by intravenous injection of rose bengal followed by focal irradiation by transluminal green light. Dietary natto-extracts supplementation containing NK of 50 or 100 CU/body was started 3 weeks before endothelial injury and then continued for another 3 weeks. Intimal thickening in animals given supplementation was significantly (P<0.01) suppressed compared with controls and the intima/media ratio in animals with 50 and 100 CU/body NK and control group was 0.09 +/- 0.03, 0.09 +/- 0.06 and 0.16 +/- 0.12, respectively. Although femoral arteries were reopened both in control animals and those treated with NK within 8 hours after endothelial injury, mural thrombi were histologically observed at the site of endothelial injury. In the control group, the center of vessel lumen was reopened and mural thrombi were attached on the surface of vessel walls. In contrast, in NK-treated groups, thrombi near the vessel wall showed lysis and most of them detached from the surface of vessel walls. In conclusion, dietary natto-extracts supplementation suppressed intimal thickening produced by endothelial injury in rat femoral artery. These effects may partially be attributable to NK, which showed enhanced thrombolysis near the vessel wall. PMID- 12850245 TI - Green tea extract inhibits angiogenesis of human umbilical vein endothelial cells through reduction of expression of VEGF receptors. AB - Epidemiological and animal studies have indicated that consumption of green tea is associated with a reduced risk of developing certain forms of cancer. However, the inhibitory mechanism of green tea in angiogenesis, an important process in tumor growth, has not been well established. In the present study, green tea extract (GTE) was tested for its ability to inhibit cell viability, cell proliferation, cell cycle dynamics, vascular endothelial growth factor (VEGF) and expression of VEGF receptors fms-like tyrosine kinase (Flt-1) and fetal liver kinase-1/Kinase insert domain containing receptor (Flk-1/KDR) in vitro using human umbilical vein endothelial cells (HUVECs). GTE in culture media did not affect cell viability but significantly reduced cell proliferation dose dependently and caused a dose-dependent accumulation of cells in the G1 phase. The decrease of the expression of Flt-1 and KDR/Flk-1 in HUVEC by GTE was detected with immunohistochemical and Western blotting methods. These results suggest that GTE may have preventive effects on tumor angiogenesis and metastasis through reduction of expression of VEGF receptors. PMID- 12850246 TI - Development of catecholaminergic neurons in the human medulla oblongata. AB - Distribution and maturation of catecholaminergic (CA) neurons have been studied by tyrosine hydroxylase immunohistochemistry in the medulla oblongata of human fetuses aged 14.5-25 weeks of gestation. Already at 14.5 weeks, CA neurons were observed in two longitudinally oriented cell clusters, one located ventrolaterally in the area of the lateral reticular and ambiguous nuclei, the other one dorsomedially forming 4 groups related to the dorsal vagal nucleus, the commissural nucleus of the vagus, the nucleus of the tractus solitarius and the area postrema. CA neurons in the area postrema were often found close to blood vessels. Scattered intermediate CA neurons were seen between these two larger clusters. CA neurons still appeared immature exhibiting bipolar morphology with only one or two short stout processes, which hardly branched. At 21 weeks, CA neurons occupied essentially the same location, but had a more mature morphology. Though still bipolar in shape, they had thinner and much longer processes which frequently branched. Both in the ventrolateral and the dorsomedial cell clusters, these processes were frequently lying close to blood vessels. At 25 weeks, CA cells had matured into multipolar neurons with long thin processes forming fine fiber networks in the ventrolateral medulla as well as around and within the dorsal vagal and solitarius nuclei. Only at this stage, a distinct CA fiber tract was seen located in the region of the tractus solitarius. Our results indicate that CA neurons in the human medulla, which are presumably involved in the control of ventilation and blood pressure, though generated rather early during development, mature relatively late. PMID- 12850247 TI - Inhibition of immunoglobulin G-catalyzed hydrogen peroxide generation by dexamethasone and piroxicam. AB - In the present study, we established a simple and physiologically acceptable in vitro assay system to measure H2O2 generated by human immunoglobulin G (IgG) and other proteins. In addition, the effects of various drugs were also tested in this method. We found that UV irradiation (280 nm) of the test solutions for 1 h at 37 degrees C produced suitable conditions to test the effects of these drugs. The test solution contained 100 microg/ml IgG in 50 mM phosphate buffer (pH 7.4), and 1% dimethylformamide (DMF), a solvent used to dissolve each drug. Phosphate anions were preferable for H2O2 generation. H2O2 concentration in the irradiated sample was determined by continuous photometric measurement of absorption (O.D.) at 340 nm for 600 sec. The decrease in O.D. was due to the oxidation of NADPH by H2O2 mediated by the glutathione redox cycle. H2O2 generation was expressed as O.D.(340 nm decrease/400 sec). IgG (100 microg/ml) generated 6-7 microM H2O2/h. With irradiation, most cytokines, proteins and enzymes failed to generate significant amounts of H2O2. The formation of H2O2 from H2O and UV light-induced singlet oxygen (1O2) was demonstrated by the inhibitory effects of 1O2 quenchers. Dexamethasone (IC50: 6 ng/ml = 1.4x10(-8) M) blocked H2O2 generation catalyzed by IgG. This action was not mediated by binding to the glucocorticoid receptor. Piroxicam (IC50: 20 ng/ml = 6.0 x 10(-6) M) and diclofenac.Na (IC50: 500 ng/ml = 1.6 x 10(-5) M), but not indomethacin, also blocked H2O2 generation. The mechanism underlying the inhibition of IgG-catalyzed H2O2 generation is not clear; however, the possibility exists that these drugs intercept, or interfere with, the approach of water molecules at the catalytic interface(s) of the IgG. PMID- 12850248 TI - Multinuclear magnetic resonance spectroscopy of high-energy phosphate metabolites in human brain following oral supplementation of creatine-monohydrate. AB - Alterations in brain high-energy phosphate metabolism, determined by in vivo magnetic resonance spectroscopy (MRS), have been reported in subjects with a number of brain disorders including major depression, schizophrenia, and substance abuse. It is not clear to what extent these changes can be modified by pharmacological or nutritional means. To address this possibility, we evaluated changes in brain chemistry that were associated with oral creatine (Cr) administration. We hypothesized that oral Cr supplementation, by increasing brain creatine and high-energy phosphate stored in phosphocreatine, would result in an increase in the creatine resonance, as measured using proton 1H-MRS, and a decrease in the beta-nucleoside triphosphate (NTP) peak and an increase in the phosphocreatine (PCr) peak, as measured by phosphorus 31P-MRS, in brain of healthy human subjects. Fifteen healthy male subjects (age=22.9+/-2.2; body mass index=22.9+/-1.7), who were without any axis I disorders or physical or neurological illness, were recruited. Ten subjects took creatine-monohydrate, 0.3 g/kg/day for the first 7 days and 0.03 g/kg/day for the next 7 days (creatine group). Five comparison subjects took equivalent amounts of sucrose as placebo (placebo group). Both 1H- and 31P-MRS scans were acquired at baseline, as well as at day 7 and day 14 of oral supplementation. 1H-MRS: Water suppressed localized spectra were acquired using a single-voxel (1.5 cm x 2 cm x 2 cm) proton MRS PRESS sequence in the left frontal lobe. 31P-MRS: Phosphorus spectral data were recorded from a 5-cm-thick axial brain slice using a short-TE slice selective spin-echo pulse sequence. The creatine group had significantly increased brain creatine levels (8.1% and 9.3%, in creatine/N-acetyl aspartate and creatine/choline ratios, respectively) compared to the placebo group over the 2 week period. The creatine group had significantly decreased beta-NTP levels (7.8%) and marginally increased PCr (3.4%) over the same period. In addition, the brain inorganic phosphate level increased over the same period in the creatine group (9.8%). The current study is the first multinuclear (1H and 31P) MRS study to evaluate changes in brain high-energy phosphate metabolism following oral creatine supplementation in healthy human subjects. These findings suggest the possibility of using oral creatine supplementation to modify brain high-energy phosphate metabolism in subjects with various brain disorders, including major depression, schizophrenia, cocaine and opiate abuse, where alterations in brain high-energy phosphate metabolism have been reported. PMID- 12850249 TI - Single-voxel proton MR spectroscopy of right versus left hippocampi in PTSD. AB - Previous magnetic resonance (MR) volume imaging and proton MR spectroscopy studies have suggested a reduction in the hippocampal size and/or neuronal/axonal density in posttraumatic stress disorder (PTSD). The lack of agreement on the laterality of the hippocampal dysfunction prompted this study. A total of 20 veterans (18 men and two women) and one female non-veteran participated in this study conducted in accordance with approved human study protocols. Six of the male veterans and the female non-veteran were without PTSD. Vietnam veterans formed a large subset of the study subjects. Single-voxel proton MR spectra were obtained from the hippocampal region bilaterally on a clinical MR scanner operating at 1.5 T. Analysis of the proton MR spectra showed a decrease in hippocampal NAA/creatine ratio in PTSD subjects significantly higher on the left than the right for the entire study group, as well as for the Vietnam subset. It was concluded that the hippocampal dysfunction in PTSD is lateralized with the left side being more impaired than the right. PMID- 12850250 TI - Neuropsychological correlates of P300 abnormalities in patients with schizophrenia and obsessive-compulsive disorder. AB - The cognitive significance of P300 abnormalities in schizophrenia and obsessive compulsive disorder (OCD) was investigated. P300 was measured by an auditory oddball paradigm, in which a series of standard tones (1000 Hz) and target tones (1500 Hz) were presented. The subject's task was to count the number of the presented target tones. Cognitive functions were evaluated by neuropsychological tests, which were chosen to be sensitive to frontal and temporal dysfunction. Twenty-two schizophrenic patients, 19 OCD patients and 21 healthy controls participated. Event-related potentials measured at 15 electrode sites, which consisted of five levels on the left-right dimension and three levels on the anterior-posterior dimension, were included in the statistical analysis. P300 amplitudes on all 15 electrode sites were significantly smaller in schizophrenic and OCD patients than in the controls. Schizophrenic patients performed poorly on almost all neuropsychological tests, while OCD patients showed impaired performance on the Rey-Osterrieth Complex Figure Test and on a controlled oral word association test. In schizophrenic patients, P300 amplitude was associated with performance on verbal memory and learning by the Luria-Nebraska Neuropsychological Battery, while for OCD patients, P300 amplitude was related to the Trail Making Test, Part B response time. These results indicate that schizophrenic patients have generalized cognitive impairments, which are substrated by a wide range of cortical dysfunctions. The major cognitive deficits observed in OCD patients were impairments of controlled attention and self guided, flexible behavior, which are mediated by the fronto-striatal system. The neurophysiological mechanisms underlying P300 abnormalities observed in schizophrenic and OCD patients are discussed. PMID- 12850251 TI - Elevated thalamic and prefrontal regional cerebral blood flow in obsessive compulsive disorder: a SPECT study. AB - Functional neuroimaging studies have pointed to a possible role of cerebral circuits involving the prefrontal and anterior cingulate cortices, the striatum, and thalamus in the pathophysiology of obsessive-compulsive disorder (OCD). Regional cerebral blood flow (rCBF) of 16 drug-free Brazilian patients with OCD and 17 healthy subjects matched for age, gender, handedness and level of education was measured with [99m-Tc] HMPAO single photon emission computed tomography. Analysis of covariance identified four regions of interest with significantly higher rCBF: the right superior and inferior frontal cortex and the right and left thalamus. Positive correlations between symptom severity measured by Clinical Global Impression scores and rCBF were found in the right and left inferior frontal lobes and in the right basal ganglia. Compulsive behavior was inversely correlated with rCBF in the right thalamus, and duration of illness correlated positively with rCBF in the right and left superior frontal lobes and with the right thalamus. The findings of this SPECT study conducted in Brazil are in agreement with prior studies and provide additional support for the involvement of prefrontal-subcortical circuits in the pathophysiology of OCD. Furthermore, the study suggests that similar brain mechanisms appear to be involved cross-culturally. PMID- 12850252 TI - Evaluation of regional cerebral blood flow changes in panic disorder with Tc99m HMPAO SPECT. AB - The objective of this study is to investigate differences in regional cerebral blood flow (rCBF) and rCBF asymmetry index values between panic disorder patients (n=22) and normal comparison subjects (n=19) using Tc99m-hexamethylpropyleneamine oxime single photon emission tomography imaging. A decrease in perfusion in the bilateral frontal regions and a relative increase in perfusion in the right medial and superior frontal regions were found. There were significant positive correlations between scores on the Panic and Agoraphobia Scale and rCBF asymmetry index values of the parietal, superior temporal and lateral temporal regions in the panic disorder patients. These correlations point to a relationship between the severity of panic disorder and relative right brain activation. Activation of the amygdala, increased CBF in the frontal region, or hyperactivation of the locus ceruleus seen in panic disorder may explain the decrease in the rCBF in the inferior frontal region. PMID- 12850253 TI - Acute effects of alcohol on hemodynamic changes during visual stimulation assessed using 24-channel near-infrared spectroscopy. AB - The purpose of this study was to evaluate the effects of alcohol on hemodynamic changes induced by visual stimulation. Ten healthy human subjects were examined using Optical Topography((R)) (Hitachi Medical Corporation: ETG-100). Each subject gradually drank 0.4 ml/kg alcohol over 10 min. Changes in oxy-hemoglobin (Hb), deoxy-Hb and total-Hb concentration were measured five times: 20 min before alcohol intake, immediately after alcohol intake, and at 20, 40 and 60 min after alcohol intake. A questionnaire was used to assess subjective feelings of alcohol. Blood-alcohol concentration (BAC) was estimated from ethanol concentration in expired air four times: immediately after alcohol intake and at 20, 40 and 60 min after alcohol intake. The visual stimulation tool was a checkerboard. It showed alternations of black and red patterns at a frequency of 8 Hz. The stimulus was displayed for 10 s after a rest of 30 s. The stimulus was repeated 10 times. Oxy-Hb concentration increased and deoxy-Hb concentration decreased during visual stimulation before and after alcohol intake, despite changes in the score of subjective feelings of alcohol and BAC. Alcohol intake does not significantly affect hemodynamic changes caused by visual stimulation in the visual cortex. PMID- 12850254 TI - The simulation of 31P NMR line shapes of lipid bilayers using an analytically soluble model. AB - Van Faassen's method for obtaining an explicit solution to a first order stochastic differential equation is applied to the simulation of 31P NMR line shapes of unoriented phospholipid bilayers in the Lalpha phase and of oriented bilayers in both the Lalpha and Lbeta' phases. The effects of the two slowest motions on the density matrix are described by the stochastic Liouville equation (SLE) which is solved analytically using the method of van Faassen. These two slowest motions are assumed to be a rotational re-orientation about the long molecular axis and a uniform wobble of this axis within a conical volume with re orientation rates characterised by correlation times tau(parallel) and tau(perpendicular) respectively. In the present work the Hamiltonian contains the intramolecular dipole-dipole interaction between the phosphorous nucleus and the four closest methylene protons of the choline headgroup, as well as the anisotropic chemical shielding interaction. Hence the contribution to relaxation from cross correlation between the dipole-dipole and anisotropic chemical shielding interactions is included. The reorientation of the headgroup is assumed to be a rotation sufficiently fast to lead to complete axially symmetric averaging of the Hamiltonian about the rotational axis (the P-O11 bond axis). Evaluation of the line shape in the present work involves only numerical integration and is therefore less computationally demanding than the large matrix inversions involved in the approaches of Campbell, Freed et al. The present theory also uses fewer parameters than that of Dufourc et al. but nevertheless results in good agreement with these authors' measurements on DMPC bilayers, using a fixed value of 10 for the ratio tau(perpendicular)/tau(parallel) in the case of the Lalpha phase. However, in contrast to Dufourc et al., we find that these correlation times are equal for the Lbeta' phase. Finally, we have simulated the decoupled powder line shapes obtained from the Lbeta' phase of DPPC by Campbell and Meirovitch. Again, we get good agreement providing tau(perpendicular)=tau(parallel). PMID- 12850255 TI - Ring-chain tautomerism in solid-phase erythromycin A: evidence by solid-state NMR. AB - Chemical shift modeling, utilizing the DFT B3LYP/D95** method, provides the spectral assignment of the 35 visible 13C resonances from the solid-phase erythromycin A dihydrate. A new resonance at 110.8ppm is observed in the high resolution 13C CP/MAS spectrum upon the application of heat or sample desiccation. With the use of the dipolar-dephasing spectral editing technique, this resonance is identified as a hemiketal carbon and the alternative hypothesis, a conformational change to the anomeric carbon of the desosamine sugar, is ruled out. Hence, the formation of a cyclic hemiketal in erythromycin A while in the solid phase is proven by solid-state NMR. The principal components of the 13C chemical-shift tensor corresponding to this hemiketal are reported. This is the first measurement of hemiketal 13C principal values. The delta11 and delta22 components are unique compared to anomeric carbon values reported in the literature. PMID- 12850256 TI - Domain sizes in heterogeneous polymers by spin diffusion using single-quantum and double-quantum dipolar filters. AB - 1H spin-diffusion experiments employing a double-quantum (DQ) dipolar filter were performed for the characterization of the microdomain structure of heterogeneous samples. For this purpose the NMR spin-diffusion process was analysed based on a model morphology of three different domains with arbitrary sizes, diffusivities, and filter efficiency. General analytical solutions for z magnetization source and sink were obtained valid for a one-dimensional lamellar morphology in the full range of spin-diffusion times. These solutions of the spin-diffusion equations were used for determining the crystalline, interface, and amorphous domain sizes in polystyrene-poly(ethylene oxide) (PS-PEO) and poly(hydroxyethylmethacrylate)-poly(ethylene oxide) (PHEMA-PEO). The DQ dipolar filter has a good efficiency for PS-PEO but is only partially efficient in filtering the signal of the mobile domains in the PHEMA-PEO diblock copolymer. The domain sizes measured by the DQ filter method are compared to those obtained using the traditional dipolar filter creating z magnetization in the mobile domains. PMID- 12850257 TI - Characterization of proteins localized to a subcellular compartment associated with an alternate secretory pathway of the malaria parasite. AB - Monoclonal antibodies recognizing proteins localized to a unique subcellular compartment within the malaria parasite are described in this report. These monoclonal antibodies recognize Plasmodium falciparum proteins of 68, 45 and 22 kDa proteins which are also conserved in rodent Plasmodium species. Co localization studies indicate that these proteins are located in a brefeldin A induced compartment which was previously proposed to be an early step in the export of proteins from the parasite into the infected erythrocyte. COPII coat proteins, Sar1p and Sec31p, and the endoplasmic reticulum-associated chaperone, BiP, all partially co-localize with the 68 and 22 kDa proteins, thus suggesting that this subcellular compartment has some similarities to the endoplasmic reticulum or that this compartment represents a domain of the endoplasmic reticulum. The 68 and 22 kDa proteins are highly soluble in non-ionic detergent and are likely to be located within the lumen of a membrane-bound compartment. These proteins found within this subcellular compartment are present throughout the blood stage from very early rings to segmenters. The results of this study further substantiate the existence of an alternate secretory pathway in the malaria parasite which plays a role in the export of proteins into the host erythrocyte. PMID- 12850258 TI - Characterization of a Rab11-like GTPase, EhRab11, of Entamoeba histolytica. AB - The Entamoeba histolytica Rab11 family of small molecular weight GTPases consists of three members, EhRab11, EhRab11B, and EhRab11C. The functions of these Rabs in Entamoeba have not been determined. Therefore, as an approach to elucidate the role of the Rab11 family of GTPases in Entamoeba, immunofluorescence microscopy was undertaken to define the subcellular localization of one member of this family, EhRab11. Under conditions of growth, EhRab11 displayed a punctate pattern in the cytoplasm of trophozoites. EhRab11 did not colocalize with markers for the Golgi apparatus, endoplasmic reticulum, pinosomes, phagosomes, or compartments formed by receptor-mediated endocytosis, suggesting that this Rab may not play a role in vesicle trafficking between these organelles. Under conditions of iron and serum starvation, EhRab11 was translocated to the periphery of the cell. The altered cellular localization was accompanied by multinucleation of the cells as well as the acquisition of detergent resistance by the cells, features that are characteristic of Entamoeba cysts. The translocation of EhRab11 to the periphery of the cell during iron and serum starvation was specific as the subcellular localizations of two other Rab GTPases, EhRab7 and EhRabA, were not altered under the same conditions. In addition, the formation of multinucleated cells by inhibition of cytokinesis was not sufficient to induce the translocation of EhRab11 to the cell periphery. Taken together, the data suggest that iron and serum starvation may induce encystation in E. histolytica and that EhRab11 may play a role in this process. Moreover, these studies are the first to describe a putative role for a Rab GTPase in encystation in Entamoeba sp. PMID- 12850259 TI - Characterisation of the merozoite surface protein-2 promoter using stable and transient transfection in Plasmodium falciparum. AB - Plasmodium falciparum merozoite surface protein (MSP)-2, is a polymorphic protein whose variable regions define two allelic families, the 3D7/IC-1 and FC27/D10 families. The gene encoding MSP-2 is located on chromosome 2 immediately 3' of the gene encoding merozoite surface protein-5 (MSP-5) with a 1096 bp intergenic region that presumably contains the MSP-2 promoter. Here we present characterization of the MSP-2 promoter using transient and stable transfection of P. falciparum. The mRNA transcription initiation site was mapped to a position 256 bp upstream of the MSP-2 translation start site. The ability of the intergenic region between MSP-5 and MSP-2 to promote the expression of chloramphenicol acetyl transferase (CAT) has been tested using a series of nested deletions in transient transfection experiments. The minimal region required for CAT expression has been defined and putative regulatory elements delineated. These nested deletions were used for heterologous expression of an FC27 family MSP-2 allele in the 3D7 allelic background in transfected 3D7 lines. In each case, the transgenic P. falciparum lines generated co-express both 3D7 and FC27 allelic forms of MSP-2 at the merozoite surface. These results have identified the functional promoter for MSP-2. PMID- 12850260 TI - Food vacuole plasmepsins are processed at a conserved site by an acidic convertase activity in Plasmodium falciparum. AB - Intraerythrocytic Plasmodium falciparum digests vast amounts of hemoglobin within an acidic food vacuole (FV). Four homologous aspartic proteases participate in hemoglobin degradation within the FV. Plasmepsin (PM) I and II are thought to initiate degradation of the native hemoglobin molecule. PM IV and histo-aspartic protease (HAP) act on denatured globin further downstream in the pathway. PM I and II have been shown to be synthesized as zymogens and activated by proteolytic removal of a propiece. In this study, we have determined that the proteolytic processing of FV plasmepsins occurs immediately after a conserved Leu-Gly dipeptidyl motif with uniform kinetics and pH and inhibitor sensitivities. We have developed a cell-free in vitro processing assay that generates correctly processed plasmepsins. Our data suggest that proplasmepsin processing is not autocatalytic, but rather is mediated by a separate processing enzyme. This convertase requires acidic conditions and is blocked only by the calpain inhibitors, suggesting that it may be an atypical calpain-like protease. PMID- 12850261 TI - Isolation and molecular cloning of a secreted hookworm platelet inhibitor from adult Ancylostoma caninum. AB - Hookworms, bloodfeeding intestinal nematodes, are a leading cause of iron deficiency anemia in the developing world. These parasites have evolved potent mechanisms of interfering with mammalian hemostasis, presumably for the purpose of facilitating bloodfeeding. Adult Ancylostoma caninum worm extracts contain an activity that inhibits platelet aggregation and adhesion by blocking the function of two cell surface integrin receptors, Glycoprotein IIb/IIIa and GPIa/IIa. Using rpHPLC, the hookworm platelet inhibitor activities have been purified from protein extracts of A. caninum. Because the two inhibitory activities co-purified through multiple chromatographic steps, have similar molecular masses and share identical N-terminal as well as internal amino acid sequence homology, it is likely that they represent a single gene product. A cDNA corresponding to the purified hookworm platelet inhibitor (HPI) protein has been cloned from adult A. caninum RNA, and the translated amino acid sequence shows significant homology to Neutrophil Inhibitory Factor and Ancylostoma Secreted Proteins, suggesting that these related hookworm proteins represent a novel class of integrin receptor antagonists. Polyclonal antibodies raised against the recombinant HPI protein recognize corresponding native proteins in A. caninum extracts and excretory/secretory products, and immunohistochemistry data have identified the cephalic glands as the major source of the inhibitor within the adult hookworm. These data suggest that HPI is secreted by the adult stage of the parasite at the site of intestinal attachment. As such, it may represent a viable target for a vaccine-based strategy aimed at interfering with hookworm-induced gastrointestinal hemorrhage and iron deficiency anemia. PMID- 12850262 TI - Biochemical and genetic evidence for a family of heterotrimeric G-proteins in Trichomonas vaginalis. AB - We have cloned a single copy gene from the human parasite Trichomonas vaginalis that encodes a putative protein of 402 amino acids with approximately 35% sequence identity to known alpha subunits of heterotrimeric G-proteins. It contains the characteristic GTP binding domains G-1 to G-5 with the key residues conserved. The new sequence has an unusual N-terminal extension of approximately 70 residues that cannot be aligned to reference G-proteins and which is characterised by proline-rich repeats. To investigate the expression and cellular localisation of the protein we produced specific antisera against a recombinant fusion protein. The antisera recognised a protein of an apparent molecular mass of 51 kDa in protein extracts from T. vaginalis and immunofluorescent microscopy established that the protein is localised to discrete endomembranes. Using a protocol designed to purify mammalian heterotrimeric G-proteins incorporating a GTPgammaS binding assay, we isolated two proteins from Trichomonas that are recognised by an heterologous GA/1 antisera raised to a peptide of the conserved G-1 domain of G-protein alpha subunits. These two proteins have an apparent molecular mass of 61 and 48 kDa, respectively, larger and smaller than the translation product of the cloned gene. Consistent with these results, the GA/1 antisera did not cross-react with the fusion protein produced from the gene we have cloned. These data suggest T. vaginalis possesses more than one heterotrimeric G-protein alpha subunit. Based on the sequence features of the cloned gene and the biochemical properties of the purified proteins, we suggest that these alpha subunits are likely to be part of classic heterotrimeric G protein complexes. PMID- 12850263 TI - The C-terminal domain of the Plasmodium falciparum acyl-CoA synthetases PfACS1 and PfACS3 functions as ligand for ankyrin. AB - Infection of erythrocytes by the malaria parasite Plasmodium falciparum results in the export of several parasite proteins into the erythrocyte cytoplasm establishing novel interactions between host and parasite proteins, particularly at the membrane skeleton that modifies both the structural and functional properties of the red cell. We present evidences that two members of the P. falciparum acyl-CoA synthetase (PfACS) family, responsible for the activation of long-chain fatty acids by thio-esterification with CoA, are transported in vesicle-like structures toward the host erythrocyte cytoplasm where they interact with the cytoskeletal protein ankyrin. Carboxyl-terminal domain (CTD) overlay studies indicated that PfACS1 and PfACS3 bind to the 78-kDa fragment of ankyrin corresponding with its spectrin-binding domain. Co-immunoprecipitation of ankyrin and PfACS1/3 indicates that at least a fraction of these proteins are physically associated in the infected erythrocytes and provide evidence for a novel specific interaction which suggest that such a binding may bring these enzymes closer to the host erythrocyte membrane where exogenous fatty acids are available. PMID- 12850264 TI - Linker scanning mutagenesis of the Plasmodium gallinaceum sexual stage specific gene pgs28 reveals a novel downstream cis-control element. AB - Protozoan parasites undergo complex life cycles that depend on regulated gene expression. However, limited studies on gene regulation in these parasites have repeatedly shown characteristics different from other eukaryotes. Within the Apicomplexa family, little is known about the mechanism of gene expression and regulation in Plasmodium spp. We have been investigating the cis-elements that control basal expression of a sexual stage specific gene in Plasmodium gallinaceum. Previously, we identified by 5' deletion analysis of a reporter construct that the 333bp upstream of the translational start site of pgs28 is sufficient for basal expression, and that the sequence between -333 and 316bp is necessary for such expression. In this report, we identified by linker scanning mutagenesis an 8-bp sequence that is essential for pgs28 transgene expression. This sequence is a target of sequence-specific nuclear factors. Primer extension studies demonstrate that, interestingly, the endogenous pgs28 transcript has two 5' ends, at -65 and +1. We suggest that this 8-bp sequence, CAGACAGC that is situated at +24 to +31 (with respect to the proximal start site), is a novel downstream promoter element in P. gallinaceum that appears to function independently of a TATA box or an Inr element. PMID- 12850265 TI - A putative Sec23 homologue of Plasmodium falciparum is located in Maurer's clefts. PMID- 12850266 TI - Retraction of "Presence of sialic acid in N-linked oligosaccharide chains and O linked N-acetylglucosamine in cruzipain, the major cysteine proteinase of Trypanosoma cruzi". [Mol Biochem Parasitol 2003; 127: 69-72]. PMID- 12850267 TI - Tissue expression and translational control of rat kynurenine aminotransferase/glutamine transaminase K mRNAs. AB - Kynurenic acid (KA) is an endogenous glutamate receptor antagonist at the level of the different ionotropic glutamate receptors. One of the enzymes responsible for the production of KA, kynurenine aminotransferase I (KATI), also catalyses the reversible transamination of glutamine to oxoglutaramic acid (GTK, EC 2.6.1.15). The enzyme exists in a cytosolic and in a mitochondrial form because of the presence of two different KATI mRNAs coding for a protein respectively with and without leader sequence targeting the protein into mitochondria. We have cloned from a phage library of rat kidney cDNA four new KATI cDNAs containing different 5' untranslated regions (UTRs). One of the transcripts (+14KATI cDNA) contains an alternative site of initiation of translation. The tissue distribution of the different transcripts was studied by RT-PCR. The study demonstrated that several KATI mRNAs are constitutively expressed in ubiquitous manner, while +14KATI mRNA is present only in kidney. The translational efficiency of the different transcripts was studied in vitro and enzymatic activities were measured in transiently transfected Cos-1 cells. Each KATI mRNA exhibits a different in vitro translational efficiency, which corresponds to different levels of KAT enzymatic activity in transfected cells. Both findings correlate with the predicted accessibility of the ribosomal binding sites of the different mRNAs. The structure of the rat KATI/GTK gene was also studied. The expression of several KATI mRNAs with different 5'UTRs represents an interesting example of transcriptional/translational control on the expression of pyridoxal phosphate (PLP)-dependent aminotransferases. PMID- 12850268 TI - Raman spectroscopy study of the interaction between biogenic polyamines and an alternating AT oligodeoxyribonucleotide. AB - The interaction between the 15-mer oligonucleotide d[A(TA)(7)].d[T(AT)(7)] and the three biogenic polyamines, putrescine, spermidine and spermine, under physiological conditions has been studied by Raman spectroscopy. Solutions containing 60 mM (phosphate) of the oligonucleotide and different polyamine concentrations ranging from 1 to 75 mM have been studied. Both natural and heavy water were used as solvents. Difference Raman spectra were computed by subtracting the sum of the separated component spectra from the experimental spectra of the complexes. The Raman data suggested that the interaction of biogenic polyamines with d[A(TA)(7)].d[T(AT)(7)] presents differences related with their sizes and electric charges. Preferential bindings through the oligonucleotide minor groove for putrescine and spermidine were proposed. Spermine would interact by both minor and major grooves, although interaction by the minor groove seems to be more favored. Main reactive sites were thymine-O2 and adenine-N3 atoms at the minor grooves and adenine-N7 and thymine-O4 at the major groove. Electrostatic attractions between the polyamine amino and oligonucleotide phosphodioxy groups were also proposed. Under our experimental conditions, no macromolecular effects on d[A(TA)(7)].d[T(AT)(7)] (changes on secondary or tertiary structures) were detected from Raman spectroscopy, contrary to what happened for GC sequences at the same experimental settings. This fact agrees with the role of the biogenic polyamines during the first steps of the macromolecular synthesis, which involve DNA opening in AT motifs. PMID- 12850269 TI - Identification of functional regulatory regions of the connexin32 gene promoter. AB - Connexin32 (Cx32) is the predominant gap junction protein expressed in adult rat hepatocytes. This study investigated transcriptional regulation of the rat Cx32 gene in MH(1)C(1) rat hepatoma cells using transient expression assays in conjunction with promoter mutagenesis and 5' nested deletion analysis. Site directed mutagenesis of the -736 and -187 hepatocyte nuclear factor-1 (HNF-1) sites, the -196 and -116 Sp1 sites, and the -729 and -329 Yin Yang 1 (YY1) sites all significantly reduced promoter activity. We have defined the contribution of each individual site to promoter activity in the intact cell. A novel upstream region of the Cx32 promoter (-1042 to -758) was cloned and shown to contain negative regulatory elements. The transcription factors HNF-1 and Sp1 have important functional roles in the transcriptional regulation of basal and cell specific Cx32 expression. The multifunctional transcription factor YY1 is also implicated. PMID- 12850270 TI - Cel6A, a major exoglucanase from the cellulosome of the anaerobic fungi Piromyces sp. E2 and Piromyces equi. AB - Anaerobic fungi possess high cellulolytic activities, which are organised in high molecular mass (HMM) complexes. Besides catalytic modules, the cellulolytic enzyme components of these complexes contain non-catalytic modules, known as dockerins, that play a key role in complex assembly. Screening of a genomic and a cDNA library of two Piromyces species resulted in the isolation of two clones containing inserts of 5.5 kb (Piromyces sp. E2) and 1.5 kb (Piromyces equi). Both clones contained the complete coding region of a glycoside hydrolase (GH) from family 6, consisting of a 20 amino acid signal peptide, a 76 (sp. E2)/81 (P. equi) amino acid stretch comprising two fungal non-catalytic docking domains (NCDDs), a 24 (sp. E2)/16 (P. equi) amino acid linker, and a 369 amino acid catalytic module. Homology modelling of the catalytic module strongly suggests that the Piromyces enzymes will be processive cellobiohydrolases. The catalytic residues and all nearby residues are conserved. The reaction is thus expected to proceed via a classical single-displacement (inverting) mechanism that is characteristic of this family of GHs. The enzyme, defined as Cel6A, encoded by the full-length Piromyces E2 sequence was expressed in Escherichia coli. The recombinant protein expressed had a molecular mass of 55 kDa and showed activity against Avicel, supporting the observed relationship of the sequence to those of known cellobiohydrolases. Affinity-purified cellulosomes of Piromyces sp. E2 were analysed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and sodium dodecyl sulfate-polyacrylamide gel (SDS PAGE) electrophoresis. A major band was detected with the molecular weight of Cel6A. A tryptic fingerprint of this protein confirmed its identity. PMID- 12850271 TI - Mutation and expression of the p53 gene during chemical hepatocarcinogenesis in F344 rats. AB - Inactivation of the p53 gene is one of the most frequent genetic alterations in carcinogenesis. We studied gene mutations, the mRNA expression of p53, and the accumulation of p53 protein in chemical hepatocarcinogenesis in rats. Samples consisting of 44 precancerous foci and 18 cancerous foci were collected by laser capture microdissection (LCM), and analyzed for mutations in rat p53 gene exons 5 8 by PCR-single-strand conformational polymorphism (PCR-SSCP). We found that 25 PCR-SSCP bands of exons 6/7 and 8 were altered in 22/62 (35.4%) LCM samples. Direct p53 gene sequencing showed that 20/62 (9 precancer, 11 cancer) (32.3%) LCM samples exhibited 34 point mutations. Ten LCM samples exhibited double or triple mutations in exons 6/7 and 8 simultaneously. A quantitative analysis of p53 mRNA showed that p53 mRNA peaked at an early stage (week 6) in the precancerous lesion, 20 times that of adjacent normal tissue, and returned to normal by week 23. Similar to precancer, p53 mRNA in cancer was five times as high as that of adjacent normal tissue at week 12, and was closer to normal at week 23. When p53 mRNA declined from a high to low, positive immunostaining for the p53 protein began to be seen in precancerous and cancerous foci, suggesting that the p53 protein had accumulated in these foci. Results show that p53 gene mutation is present in initial chemical hepatocarcinogenesis and p53 mRNA concentration is clearly elevated before gene mutation. Once the p53 gene has mutated, mRNA concentration progressively declines, suggesting that mutation leads to inactivation of the p53 gene. PMID- 12850272 TI - Differential translation of TOP mRNAs in rapamycin-treated human B lymphocytes. AB - TOP mRNAs (contain a 5' terminal oligopyrimidine tract) are differentially translated in rapamycin-treated human B lymphocytes. Following rapamycin treatment, ribosomal protein (rp) and translation elongation factor TOP mRNAs were translationally repressed, whereas hnRNP A1 TOP mRNA was not. Poly(A) binding protein (Pabp1) TOP mRNA was translationally repressed under all conditions tested. To investigate the mechanism involved, chimeric mRNAs containing the hnRNP A1 5' untranslated region (UTR) linked to the human growth hormone (hGH) reporter were analyzed. Wild-type hnRNP A1 construct mRNA behaved similarly to endogenous hnRNP A1, whereas a single mutation (guanosine to cytidine) within the TOP element resulted in increased translational regulation. These results suggest that TOP mRNA translation can be modulated and that all TOP mRNAs are not translated with equal efficiency. PMID- 12850273 TI - Cloning and characterization of an eukaryotic initiation factor-2alpha kinase from the silkworm, Bombyx mori. AB - Eukaryotic initiation factor 2alpha (eIF-2alpha) kinases are involved in the translational regulations that occur in response to various types of environmental stress, and play an important role in the cellular defense system operating under unfavorable conditions. The identification of additional eIF 2alpha kinases and the elucidation of their functions are necessary to understand how different eIF-2alpha kinases can specifically respond to distinct stimuli. Here, we report a novel eIF-2alpha kinase, termed BeK, from the silkworm, Bombyx mori. This gene encodes 579 amino acids and contains all 11 catalytic domains of protein-serine/threonine kinases. Most notably, it contains an "Ile-Gln-Met-Xaa Xaa-Cys" motif, which is highly conserved from yeast to mammalian eIF-2alpha kinases. BeK does not show any significant homology in the NH(2) terminal regulatory domain, suggesting a distinct regulatory mechanism of this novel eIF 2alpha kinase. BeK is ubiquitously expressed in the various tissues throughout the final larval stage. Importantly, BeK is activated in Drosophila Schneider cells following heat shock and osmotic stress, and activated-BeK has been shown to phosphorylate an eIF-2alpha subunit at the Ser(50) site. However, other forms of stress, such as immune stress, endoplasmic reticulum stress and oxidative stress, cannot significantly elicit BeK activity. Interestingly, the baculovirus gene product, PK2, can inhibit BeK enzymatic activity, suggesting that BeK may be an endogenous target for a viral gene product. Taken together, these data indicate that BeK is a novel eIF-2alpha kinase involved in the stress response in B. mori. PMID- 12850274 TI - Genomic organization, chromosomal localization and adipocytic expression of the murine gene for CORS-26 (collagenous repeat-containing sequence of 26 kDa protein). AB - The murine gene for CORS-26 shows striking homologies to the adipocyte-specific secretory protein adiponectin (belonging to the newly discovered C1q/TNF molecular superfamily) and its expression has been reported to be restricted to fibroblasts, cartilage and kidney. However, the present data demonstrate specific induction of CORS-26 mRNA expression in hormonally differentiated 3T3-L1 adipocytes, but not in preadipocytes. Furthermore, CORS-26 mRNA expression could be demonstrated in human synovial adipocytes of the knee by in situ hybridization. Since the genes for CORS-26 and adiponectin are homologous for their COOH-terminal globular domain and of their N-terminal collagenous domain, they might have originated by divergence from an innate mesenchymal precursor molecule directing the development of myocytes, adipocytes and chondrocytes from a mesenchymal stem cell. Here, the complete genomic organization with exon/intron boundaries together with exon-specific primer combinations are presented. Additionally, approximately 1 kb of the TATA-box-containing promoter region was cloned and analyzed for putative transcription factor binding sites. The chromosomal localization of the murine CORS-26 gene was mapped to mouse chromosome 15 A2 by fluorescence in situ hybridization (FISH). Since the linkage loci for proteoglycan-induced arthritis and MRL/lpr arthritis in mice have been mapped to that chromosomal region, CORS-26 might represent the underlying mechanism of disease. The present data provide the basis for further investigation of the CORS-26 gene regulation in the context of mesenchymal tissue development, chondrocyte/adipocyte function and bone or skeletal disease. PMID- 12850275 TI - Genomic structure and expression analysis of the gene encoding a silkworm basic Kunitz-type chymotrypsin inhibitor. AB - Kunitz-type chymotrypsin inhibitor CIb1 of silkworm Bombyx mori is a basic peptide consisting of 62 amino acid residues. To elucidate the mechanisms of transcriptional regulation of CIb1 gene expression, we cloned it for genomic structure analysis. CIb1 cDNA was used as a probe to screen a BAC sub-library. One positive clone containing the upstream sequences was isolated and the sequence result showed that CIb1 gene consists of three exons spaced by two introns. In the 5'-flanking region, consensus TATA and CCAAT boxes were identified. Other binding sites for transcription factors such as NF-kappaB, GATA, C/EBP, COUP-TF/HNF-4, RORalpha1, SRY, and HOXA3 were also detected. Southern blot analysis suggested a single copy of CIb1 gene in the silkworm genome. Northern blot analysis indicated that the expression of CIb1 gene is transcriptionally regulated during development and is apparently tissue-specific. The CIb1 mRNA was detected in fat body, ovary, trachea, and skin. We furthermore investigated the CIb1 expression profiles after LPS and E. coli injection. The fluctuations of CIb1 transcript in challenged larvae confirm our proposal that CIb1 is an immune responsible gene. According to our data, we discussed the transcriptional factors putatively responsible for the physiological role of CIb1 in the silkworm hemolymph. PMID- 12850276 TI - The luteinizing hormone receptor: influence of buffer composition on ligand binding and signaling of wild type and mutant receptors. AB - There is evidence that ligand binding to and ligand-mediated signaling by the luteinizing hormone receptor (LHR) are influenced by buffer conditions, including ionic type and strength, an issue that becomes important in comparing functional parameters obtained on receptor mutants under different conditions. In order to study this phenomenon, we performed binding (kinetic and saturation) and signaling studies of human chorionic gonadotropin (hCG) with wild type (wt) LHR and several mutants expressed in COS-7 cells using two common buffer systems. One buffer was of low ionic strength and contained a low concentration of Na+, while the other had a near-physiological concentration of Na+. Emphasis was placed on mutations of two amino acid residues in the hinge region of the ectodomain (E332 and D333). It was found that the buffer of higher ionic strength, primarily from Na+, led to an increase of about 4-fold in the Kd of hCG binding to wt and mutant LHRs. The reduced binding affinities were attributable to a comparable reduction in the rate constants of association, with no significant differences in the calculated rate constants of dissociation in the two buffers. Analysis of the signaling properties of these mutants showed that, when corrected for the amount of hCG bound under the conditions of the signaling assay, the maximal ligand mediated cAMP produced in cells maintained in the buffer of low ionic strength was comparable for wt LHR and the mutants, only the D333A mutant being somewhat elevated. In the buffer of higher ionic strength, however, the response by wt LHR was significantly greater than that of the mutants. These results show that E332 and D333 are important in hormone-mediated signaling, but only in the buffer of higher Na+ concentration. In addition to mutants of these two residues, the buffer of higher ionic strength also led to reduced binding to a number of mutants throughout the receptor. Since these mutants included additional replacements in the ectodomain and transmembrane helices 6 and 7, the general nature of the buffer effect on wt and mutant LHRs suggests that electrostatic effects are contributing to ligand binding and/or that the LHR ectodomain may exist in two conformations, one being more accessible to ligand at reduced ionic strength. PMID- 12850277 TI - Non-covalent interaction of ubiquitin with insulin-degrading enzyme. AB - Insulin-degrading enzyme (IDE) is a metalloprotease implicated in insulin degradation and suggested to have a variety of additional functions, including the clearance of amyloid beta peptides of Alzheimer's disease. Little is known about endogenous proteins that may interact with and modulate IDE's activity in the cell. We purified and characterized two proteins from mouse leukemic splenocytes that interact with IDE and inhibit its insulin-degrading activity. A protein of 14 kDa was similar to a competitive IDE inhibitor reported previously. The major inhibitor was identified by amino acid sequencing as ubiquitin, a protein that is post-translationally covalently attached to other intracellular proteins and regulates diverse cellular processes. Ubiquitin inhibited insulin degrading activity of IDE and diminished crosslinking of 125I-insulin to IDE in a specific, concentration-dependent, reversible, and ATP-independent manner. Ubiquitin did not affect the crosslinking of 125I-insulin to insulin receptors or of 125I-atrial natriuretic peptide (ANP) to its receptor guanylate cyclase-A. These findings suggest a novel role for ubiquitin or perhaps proteins with ubiquitin-like domains in regulating the function of IDE. PMID- 12850278 TI - Analysis of human choriogonadotropin core 2 o-glycan isoforms. AB - Measurements of human choriogonadotropin (hCG) isoforms containing core 2 o glycans may be useful for diagnosis of Down Syndrome pregnancies and trophoblastic disease. As shown here, this isoform is also present in pituitary extracts, early pregnancy urine, and urine of postmenopausal women. Although, measurements of hCG isoforms may be useful in several clinical settings, this remains to be determined due to the lack of suitable standards and the difficulties of comparing data obtained in different laboratories. Here, we report that monoclonal antibodies B152 and CTP104 recognize the third (Ser132) and fourth (Ser138) o-glycans, respectively, in the carboxyterminal portion of the hCG beta-subunit. The proximity of these sites prevents B152 and CTP104 from binding simultaneously to isoforms containing core 2 o-glycans. Unlike B152, which binds only the core 2 isoform, CTP104 recognizes both glycan moieties. By measuring hCG with CTP104 in the presence and absence of B152, one can quantify both isoforms using the same readily available standard. PMID- 12850279 TI - Liver-specific IGF-I gene deficient mice exhibit accelerated diabetes in response to streptozotocin, associated with early onset of insulin resistance. AB - Liver-specific IGF-I gene deficient (LID) mice exhibit pancreatic islet hyperplasia and insulin resistance. To clarify their causal relationship, we studied age-dependent changes in these two aspects and the response to beta-cell damage caused by streptozotocin in adult mice. As a result, the onset of insulin resistance in LID mice was detectable as early as 1-month of age, while hyperinsulinemia was developed after a significant delay at 2.5-month. Upon streptozotocin administration, control mice exhibited significant hyperglycemia after 9 days, and glucose levels continued to rise at 12-15 days. LID mice developed diabetes much more rapidly, with hyperglycemia after 6 days and higher glucose levels up to 15 days. They also exhibited significant weight loss and 6/19 died. Serum insulin assay, insulin mRNA analysis and immunohistochemistry revealed that the more severe diabetes in LID mice was not due to more damage to their beta-cells. Thus LID mice are more sensitive to streptozotocin-induced beta cell damage, due to a primary defect in insulin responsiveness. The pancreatic islet hyperplasia observed in these mice seems to represent a compensatory response to insulin resistance, therefore, offers no protection against beta-cell damage. PMID- 12850280 TI - Cell signalling of the GLP-1 action in rat liver. AB - GLP-1, incretin with insulin-independent antidiabetic properties, is insulinomimetic upon glucose metabolism in extrapancreatic tissues, acting through specific receptors not associated to adenylate cyclase activation. We investigated the role of enzymes mediating insulin actions, in the GLP-1-induced glycogen synthase a activation in rat hepatocytes. GLP-1, like insulin, activates PI3K/PKB, p70s6k, p44 and p42 MAP-kinase. Wortmannin (PI3K/PKB inhibitor) blocked the stimulatory action of insulin on glycogen synthase a and reduced that of GLP 1; rapamycin (p70s6k inhibitor) was ineffective and PD98059 (MEK/MAPK inhibitor) decreased only the insulin effect; okadaic acid (PP-2A inhibitor) was ineffective, while TNFalpha (PP-1 inhibitor) blocked the action of insulin and reduced that of GLP-1; H-7 or Ro 31-8220 (PKC inhibitors) decreased the GLP-1 effect, while only H-7 reduced that of insulin. The activation of PI3K/PKB, PKC and PP-1, but not PP-2A, seems to mediate the GLP-1 stimulatory action on glycogen synthase a in rat hepatocytes, while MAPKs and p70s6k could participate in other GLP-1 effects. PMID- 12850281 TI - Prostate cancer cell type-specific regulation of the human PTHrP gene via a negative VDRE. AB - Parathyroid hormone-related protein (PTHrP) is expressed by prostate cancer cells. Since PTHrP increases the growth and enhances the osteolytic effects of prostate cancer cells, it is important to control the level of PTHrP expression in these cells. We show that 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and its non calcemic analogue, EB1089, suppress PTHrP mRNA and protein levels in the human prostate cancer cell lines PC-3 and LNCaP. The human PTHrP gene contains a sequence element homologous to the negative vitamin D response element within the parathyroid hormone gene. This DNA sequence (nVDRE(hPTHrP)) bound the vitamin D receptor (VDR) present in nuclear extracts from both PC-3 and LNCaP cells. However, when cloned upstream of the SV40 promoter and transiently transfected into PC-3 and LNCaP cells, nVDRE(hPTHrP) downregulated promoter activity in response to 1,25(OH)2D3 or EB1089 treatment in LNCaP, but not in PC-3, cells. These results may help to explain why some prostate cancers appear to be refractory to treatment with vitamin D analogues. PMID- 12850282 TI - Prolactin-like protein-A is a functional modulator of natural killer cells at the maternal-fetal interface. AB - Natural killer (NK) cells are the predominant lymphocytes present in healthy rodent and human implantation sites. In the rat, the expansion, differentiation and subsequent migration of NK cells away from the developing chorioallantoic placenta coincide with the expression of a novel pregnancy- and trophoblast cell specific cytokine, prolactin (PRL)-like protein A (PLP-A). PLP-A specifically binds to uterine NK cells but does not appear to utilize receptor systems for PRL. In the present report, we show that PLP-A interactions with NK cells are not mediated by receptors utilized by known modulators of NK cell function, including interleukin-2, interleukin-7, interleukin-12, and interleukin-15 (IL-15). Uterine NK cells respond to PLP-A or IL-15 with an increase in intracellular calcium mobilization. In contrast, PLP-A, unlike IL-15, effectively suppresses the ability of NK cells to produce interferon-gamma (IFNgamma), a key mediator of NK cell function. Placental PLP-A expression is reciprocal to mesometrial decidua expression of IFNgamma. Increased expression of PLP-A by the placenta coincides with the decline of IFNgamma content in the mesometrial decidua adjacent to the placenta. In summary, trophoblast cell-derived PLP-A contributes to the regulation of NK cells at the maternal-fetal interface to ensure appropriate embryonic growth and development. PMID- 12850283 TI - Prolactin regulates adenylyl cyclase and insulin secretion in rat pancreatic islets. AB - A role for prolactin (PRL) in the regulation of adenylyl cyclase (AC), cyclic AMP (cAMP) formation and insulin secretion was studied in isolated rat pancreatic islets cultured for 4 days at 5.5 mM glucose in the absence (control) or presence of PRL (500 ng/ml). In PRL-treated islets, stimulation by glucose (8 mM), carbamylcholine chloride (CCh) and phorbol dibutyrate increased cAMP levels 40, 89, and 151%, respectively, above similarly stimulated control islets without PRL. Moreover, insulin secretion in PRL-treated islets was more than doubled in response to 8 mM glucose plus glucagon-like peptide 1 compared with control islets. PRL also increased protein kinase C (PKC) activity in cultured islets. When islets were cultured at an insulin secretion desensitizing concentration of glucose (11 mM) for 4 days, there was a decrease in forskolin-stimulated cAMP production. However, the presence of PRL with 11 mM glucose prevented the glucose induced decrease in cAMP production. Insulin secretion in response to 17 mM glucose was also higher (P<0.02) in islets cultured with 11 mM glucose plus PRL compared with islets cultured with 11 mM glucose alone. Islet AC types -III, -V, and -VI mRNA levels increased relative to 18s rRNA following PRL treatment. In contrast, culture at 11 mM glucose decreased relative AC-III, -V and -VI mRNA levels by as much as 50%. Culture with PRL prevented the decrease in AC expression during islet culture with 11 mM glucose, and the mRNA levels remained similar to control islets cultured at 5.5 mM glucose. Thus, PRL not only increased islet AC expression and activity and insulin secretory responsiveness, but also protected islets from chronic glucose-induced inhibition of these beta cell activation parameters. PMID- 12850284 TI - IRS-3 inhibits IRS-2-mediated signaling in pancreatic beta-cells. AB - IRS-2 plays an important role in the control of pancreatic beta-cell growth, however it is unclear if other IRS family members are also involved. Using recombinant adenoviruses, IRS-1, -2 and -3 expression was varied in the beta-cell line, INS-1. Increased IRS-1 expression had no appreciable effect on beta-cell growth. However, increased IRS-2 expression augmented glucose/IGF-1 induced beta cell growth mitogenesis and decreased apoptosis due to glucose-deprivation. In contrast, increased IRS-3 expression significantly inhibited mitogenesis and increased apoptosis. IRS-3 was intransiently located to the beta-cell plasma membrane, and appeared to be inert in terms of IGF-1 induced signaling. However, increased IRS-3 expression blocked glucose/IGF-1 induced IRS-2 translocation from the cytosol to the plasma membrane, dampening IRS-2/IGF-1R interaction and subsequent activation of the PI3K/PKB/GSK3 signaling pathway. In contrast, glucose/IGF-1 induced Erk-1/-2 and p70S6K activation were unaffected by IRS-3. These data emphasize the importance of IRS-2/PI3K/PKB signal transduction for beta-cell growth and survival. PMID- 12850285 TI - Identification of hemopexin as a GH-regulated gene. AB - A cDNA library from the liver of a growth hormone (GH)-treated hypophysectomized rat was constructed and screened for GH-inducible genes (GIGs). Three cDNAs specific for putative GIG mRNAs (GIG-3, -7 and -12) were isolated and, when sequenced, were found to be homologous to portions of rat hemopexin, a Class 2 acute-phase gene. Hemopexin is an essential heme scavenger produced primarily in the liver, which upon binding to free heme, transports it to the liver where the heme iron is re-utilized. Hemopexin has not been previously described as being GH responsive. GIG-3 and GIG-12 encode overlapping portions of the entire coding sequence starting within a few hundred base pairs from the 5' end of the hemopexin mRNA, and GIG-7 encodes the 3'-most end of the hemopexin mRNA. Northern analysis and ribonuclease protection assays of RNA from livers of control rats using the cDNA probes demonstrated a major transcript of approximately 2.0 kb. The hemopexin mRNA was low or undetectable in livers of hypophysectomized rats. Daily treatment with bovine growth hormone (bGH) for 10 days restored hemopexin mRNA to levels comparable or greater than that of intact rats. GH-dependence in cultured rat H4IIE hepatoma cells was then examined. Using hemopexin cDNA probes (GIG-3, -7, and -12) we identified a mRNA on Northern blots, which increased in concentration following bGH, compared with untreated cells. When measured by ribonuclease protection assay, a maximal increase in hemopexin mRNA concentration was obtained following 4-6 h of bGH administration. We conclude that hemopexin is a GH-inducible gene in rat liver in vivo and in cultured rat hepatoma cells. PMID- 12850286 TI - Regulation of hemopexin transcription by calcium ionophores and phorbol ester in hepatoma cells. AB - Hemopexin (Hx) is an acute-phase hepatic protein, whose transcription is upregulated by IL-6. The transcription rate of Hx was found to be increased 11 fold by the calcium ionophore A23187, 25-fold by the calcium ionophore ionomycin, and 4-5-fold by phorbol 12-myristate 13-acetate (PMA) in serum-starved H4IIE rat hepatoma cells. Insulin did not affect the transcription rate of Hx. These findings are consistent with involvement of intracellular calcium concentrations and activation of protein kinase C (PKC) action in the regulation of Hx. PMID- 12850287 TI - Functional relationship of serine 90 phosphorylation and the surrounding putative salt bridge in bovine prolactin. AB - The biological activity of bovine prolactin (PRL) is reduced by in vivo phosphorylation of serine 90 (S90) that is located within a putative N+4 salt bridge (R89 and D93). We substituted hydrophobic, polar, or acidic residues for S90 and/or replaced members of the putative R89/D93 salt bridge to determine if a functional relationship between the putative salt bridge and the phosphorylation could be observed. At position 90 the bulk of the residue was the most important factor in modulating biological activity in either the rat Nb2 cell bioassay or PRL receptor binding. Charge played a smaller role. Replacement of either partner of the salt bridge reduced both biological and binding activities indicating the presence of a salt bridge at this position. The combination of replacing a salt bridge member and substituting glutamic acid at S90 produced greater than additive changes in our experimental endpoints, indicating a functional coupling between the salt bridge and phosphorylation site. We interpret the data to indicate that either in vivo phosphorylation or specific mutations that destabilize the salt bridge impairs biological activity. PMID- 12850289 TI - P2X and P2Y purinoceptor expression in pancreas from streptozotocin-diabetic rats. AB - The expression of the nucleotide receptors P2X1, P2X2, P2X7, P2Y1, P2Y2 and P2Y4, in the pancreas of the streptozotocin-induced diabetic rat was investigated using immunohistochemistry. In diabetic animals, P2X7 receptor expression, normally located in the outer periphery of the islet, was increased and located inside the islet. Double-labelling experiments, using antibodies raised against insulin, somatostatin and glucagon, showed, for the first time, an increase in immunostaining for P2X7 receptors on islet glucagon-containing alpha cells (which had migrated to the interior), while no P2X7 receptors were found in beta and delta cells. P2Y1 receptors were present in intra-islet capillaries, while P2Y4 receptors were found on both alpha and beta cells. P2Y1 and P2Y2 receptor expression was also found in pancreatic duct cells and P2X1, P2X2, P2Y1 and P2Y2 receptors were identified in small blood vessels. PMID- 12850288 TI - The association of arrestin-3 with the follitropin receptor depends on receptor activation and phosphorylation. AB - We have recently shown that the binding of arrestin-3 to the lutropin receptor (LHR) is dependent mostly on receptor activation rather than on phosphorylation. The experiments presented here were designed to test the involvement of these two events in the association of arrestin-3 with the closely related follitropin receptor (FSHR). Activation of the FSHR leads to the phosphorylation of residues in the first and third intracellular loops. Mutation of the phosphorylation sites in the third intracellular loop of the rat (r) FSHR partially reduces phosphorylation but has no effect on arrestin-3 association. Mutation of the phosphorylation sites in the first intracellular loop abolishes phosphorylation and arrestin-3 association. Dominant-negative mutants of G protein-coupled receptor kinase (GRKs) 2 and 6 inhibit rFSHR phosphorylation to the same extent but only the dominant-negative mutant of GRK2 inhibits arrestin-3 association. Two mutations of the rFSHR (D389N and Y530F) that impair activation and abolish phosphorylation also impair arrestin-3 binding. GRK2 restores the phosphorylation of both mutants but it restores arrestin-3 association only to the D389N mutant. We conclude that, in contrast to the data obtained with the LHR, the association of arrestin-3 with the FSHR is dependent on receptor phosphorylation. The phosphorylation of the third intracellular loop residues is not needed for arrestin-3 association, however. PMID- 12850290 TI - Ets-1 activates parathyroid hormone-related protein gene expression in tumorigenic breast epithelial cells. AB - Parathyroid hormone-related protein (PTHrP) is produced by many tumors not associated with humoral hypercalcemia, including breast cancers. In this study, we used three human immortalized mammary epithelial cell lines that differ in tumorigenicity and PTHrP expression. Using RT-PCR we investigated 5' and 3' alternative splicing of PTHrP transcripts and promoter usage in the lines. Increased levels of P3-derived transcripts and the 1-139 mRNA isoform were observed in the most tumorigenic cell line. Transient transfection experiments identified elements close to P3 promoter that appeared to account for a portion of differential PTHrP expression among the three cell lines. Using site-directed mutagenesis, a previously described Ets-1/Sp1 binding site upstream of P3 was determined to be crucial for full activity of this promoter. RT-PCR and western blot evaluation of Ets family member expression found that Ese-1 was present in all three lines, but that appreciable levels of Ets-1 protein were present exclusively in the most tumorigenic line. Cotransfection of Ets-1 expression vectors activated PTHrP reporter constructs in the most tumorigenic line but not in the other cell lines. These findings suggest a potential mechanism by which PTHrP transcription may be regulated as a consequence of events that promote tumorigenic behavior in breast epithelial cells. PMID- 12850291 TI - Molecular cloning of estrogen receptor alpha and expression pattern of estrogen receptor subtypes in male and female goldfish. AB - Estrogen plays an important role in the regulation of gonadotropin production in vertebrates. In this study, we isolated the estrogen receptor (ER) alpha cDNA from the goldfish pituitary. Primers for ERalpha were designed based on the similarity of selected regions (C and E domains) of known ER genes. Full-length cDNA sequence for ERalpha was determined by 3' and 5' RACE procedures. ERalpha cDNA clone was found to contain 2087 nucleotides including an open reading frame that encodes 564 amino acids, with a molecular weight of 62.8 kDa. We also cloned ERbeta-1 and ERbeta-2 from the published information and investigated the expression pattern of these ER subtypes in a variety of tissues in male and female goldfish by reverse transcriptase-polymerase chain reaction (RT-PCR). Significant variations in the relative expression of ERalpha, ERbeta-1 and ERbeta 2 were observed in different tissues in male and female goldfish. Pituitary was found to have the highest expression level of ERalpha in both male and female goldfish. Significantly, lower levels of ERalpha expression were observed in the brain, ovary, testis, liver, muscle, heart and intestine. Ovary and testis were found to have higher transcript levels of ERbeta-1 with much lower levels in the brain, pituitary, liver, muscle and heart. The ERbeta-2 was found to be expressed strongly in the pituitary followed by intestine with lower expression in other tissues. The present study provides molecular characterization of ERalpha, and information on tissue specific distribution of different ER subtypes in male and female goldfish. PMID- 12850293 TI - An improved stereologic method for three-dimensional estimation of particle size distribution from observations in two dimensions and its application. AB - Single enzyme-altered hepatocytes; altered hepatic foci (AHF); and nodular lesions have been implicated, respectively in the processes of initiation, promotion, and progression in rodent hepatocarcinogenesis. Qualitative and quantitative analyses of such lesions have been utilized both to identify and to determine the potency of initiating, promoting, and progressor agents in rodent liver. Of a number of possible parameters determined in the study of such lesions, estimation of the number of foci or nodules in the liver is very important. The method of Saltykov has been used for estimating the number of AHF in rat liver. However, in practice, the Saltykov calculation has at least two weak points: (a) the size class range is limited to 12, which in many instances is too narrow to cover the range of AHF data obtained; and (b) under some conditions, the Saltykov equation generates negative values in several size classes, an obvious impossibility in the real world. In order to overcome these limitations in the Saltykov calculations, a study of the particle size distribution in a wide-range, polydispersed sphere system was performed. A stereologic method, termed the 25F Association method, was developed from this study. This method offers 25 association factors that are derived from the frequency of different-sized transections obtained from transecting a spherical particle, thus expanding the size class range to be analyzed up to 25, which is sufficiently wide to encompass all rat AHF found in most cases. This method exhibits greater flexibility, which allows adjustments to be made within the calculation process when NA((k,k)), the net number of transections from the same size spheres, was found to be a negative value, which is not possible in real situations. The reliability of the 25F Association method was tested thoroughly by computer simulation in both monodispersed and polydispersed sphere systems. The test results were compared with the original Saltykov method. We found that the 25F Association method yielded a better estimate of the total number of spheres in the three-dimensional tissue sample as well as the detailed size distribution information. Although the 25F Association method was derived from the study of a polydispersed sphere system, it can be used for continuous size distribution sphere systems. Application of this method to the estimation of parameters of preneoplastic foci in rodent liver is presented as an example of its utility. An application software program, 3D_estimation.exe, which uses the 25F Association method to estimate the number of AHF in rodent liver, has been developed and is now available at the website of this laboratory. PMID- 12850294 TI - 'Minimum average risk' as a new peak-detection algorithm applied to myofibrillar dynamics. AB - We present a new peak-detection algorithm based on the method of 'minimum average risk' proposed by Kolmogorov and developed for signal processing in various fields. In this method, translations of features within a signal scan are quantified by minimizing the integrated pointwise product of each scan relative to the first derivative of the immediately previous scan. We have adapted this method for use in a new algorithm to monitor dynamic changes of sarcomere length in single myofibrillar sarcomeres of striated muscles, but the algorithm can also be used more generally for peak localization. We find that this method results in sub-nanometer precision and higher signal-to-noise ratio than current methods. At an equal noise level, the RMS deviation of the minimum average risk algorithm was 1.3 times lower than that of the center of mass method with modeled data and 3-4 times lower with actual data. PMID- 12850295 TI - Ordered subsets Bayesian tomographic reconstruction using 2-D smoothing splines as priors. AB - The ordered subsets expectation maximization (OS-EM) algorithm has enjoyed considerable interest for accelerating the well-known EM algorithm for emission tomography. The OS principle has also been applied to several regularized EM algorithms, such as nonquadratic convex minimization-based maximum a posteriori (MAP) algorithms. However, most of these methods have not been as practical as OS EM due to their complex optimization methods and difficulties in hyperparameter estimation. We note here that, by relaxing the requirement of imposing sharp edges and using instead useful quadratic spline priors, solutions are much easier to compute, and hyperparameter calculation becomes less of a problem. In this work, we use two-dimensional smoothing splines as priors and apply a method of iterated conditional modes for the optimization. In this case, step sizes or line search algorithms necessary for gradient-based descent methods are avoided. We also accelerate the resulting algorithm using the OS approach and propose a principled way of scaling smoothing parameters to retain the strength of smoothing for different subset numbers. Our experimental results show that the OS approach applied to our quadratic MAP algorithms provides a considerable acceleration while retaining the advantages of quadratic spline priors. PMID- 12850296 TI - Lossy compression of eye movement and auditory brainstem response signals. AB - Eye movement and auditory brainstem response signals recorded for balance and hearing investigations were used as a medical test battery for several types of lossy compression techniques. These signals are associated with the function of the ears. The former signals are used to assess the balance problems (especially vertigo) of a subject and the latter his or her hearing problems. New technique is also presented based on successive approximation quantization. The effect of information loss on medical parameters computed from the signals in the course of compression was evaluated for brainstem response signals. It is important to ensure that lossy compression techniques of these biomedical signals do not impair medical parameter values computed from the signals. PMID- 12850297 TI - Integration of CT, CAD system and finite element method to investigate interfacial stresses of resin-bonded prosthesis. AB - The question of whether prosthesis/abutment tooth interface debonding is associated with inappropriate occlusal force is investigated in this study. A new modeling approach was employed to perform the interfacial stress analyses. Solid models of resin-bonded (RB) prosthesis and abutment teeth were constructed by stacking serious section contours that were obtained from CT images. A 3-D finite element (FE) model of RB prosthesis/abutment teeth was generated in a CAD system after assembling and meshing procedures. An in-house program was developed to combine the FE package (ANSYS) to calculate the interfacial (normal and shear) stresses at the prosthesis/molar interface with the bonding and debonding conditions. After 10 different occlusal force(s) evaluations, three initial opening gaps at the distal margin of the retainer were assumed to examine the possible interface debonding mechanism under the worst loading case. The results indicated that a more accurate FE model of the posterior RB prosthesis/abutments could be generated through combining several computer-aided techniques. The maximum interfacial stresses were obtained when the occlusal force was applied on the buccal slope of the distolingual cusp of the molar. For interface debonding simulations, peak normal and shear stresses were found near the debonding areas and stress values increased gradually with small to large initial opening gaps. From these results, prosthesis/abutment tooth interfacial fatigue damage might arise or accelerate the interface to debond under adequate bonding or initial gap opening conditions after long-term repeated inappropriate occlusal force actions. PMID- 12850298 TI - Towards knowledge-based systems in clinical practice: development of an integrated clinical information and knowledge management support system. AB - Given that clinicians presented with identical clinical information will act in different ways, there is a need to introduce into routine clinical practice methods and tools to support the scientific homogeneity and accountability of healthcare decisions and actions. The benefits expected from such action include an overall reduction in cost, improved quality of care, patient and public opinion satisfaction. Computer-based medical data processing has yielded methods and tools for managing the task away from the hospital management level and closer to the desired disease and patient management level. To this end, advanced applications of information and disease process modelling technologies have already demonstrated an ability to significantly augment clinical decision making as a by-product. The wide-spread acceptance of evidence-based medicine as the basis of cost-conscious and concurrently quality-wise accountable clinical practice suffices as evidence supporting this claim. Electronic libraries are one step towards an online status of this key health-care delivery quality control environment. Nonetheless, to date, the underlying information and knowledge management technologies have failed to be integrated into any form of pragmatic or marketable online and real-time clinical decision making tool. One of the main obstacles that needs to be overcome is the development of systems that treat both information and knowledge as clinical objects with same modelling requirements. This paper describes the development of such a system in the form of an intelligent clinical information management system: a system which at the most fundamental level of clinical decision support facilitates both the organised acquisition of clinical information and knowledge and provides a test-bed for the development and evaluation of knowledge-based decision support functions. PMID- 12850299 TI - Model validation software for classification models using repeated partitioning: MVREP. AB - The process of assessing the prediction ability of a computational model is called model validation. For models predicting a categorical response, the prediction ability is usually quantified by prediction measures such as sensitivity, specificity, and accuracy. This paper presents a software Model Validation using Repeated Partitioning (MVREP) that implements a computer intensive, nonparametric approach to model validation, which we call the re partitioning method. MVREP, developed using the SAS Macro language, repeats the process of randomly partitioning a dataset and subsequently performing standard model validation procedures, such as cross-validation, a large number of times and generates the empirical sampling distributions of prediction measures. The means of the sampling distributions serve as the point estimates of prediction measures of the model. The variances of the sampling distributions provide a direct assessment of variability for the point estimates of prediction measures. An example is presented using a mouse developmental toxicity chemical dataset to illustrate how the software can be used for the assessment of structure-activity relationships models. PMID- 12850300 TI - Photocontact dermatitis and chloracne: two major occupational and environmental skin diseases induced by different actions of halogenated chemicals. AB - Among occupational and environmental disorders, contact or photocontact dermatitis and an acneiform eruption are two major skin disorders. Photocontact dermatitis was historically caused by various halogenated salicylanilides, while the acne is induced by halogenated aromatic hydrocarbons and thus called chloracne. Therefore, it should be noted that halogenated chemical compounds are important causative agents in the occupational and environmental medicine. In photocontact dermatitis, photoconjugation of epidermal cells with a photohaptenic halogenated chemical is the initial step. Langerhans cells serve as antigen presenting cells and T cells sensitized by photoantigen-bearing Langerhans cells induce this photosensitivity. On the other hand, in chloracne, halogeneted hydrocarbons render keratinocytes of the outer root sheath and sebaceous duct hyperplastic. The dilated infundibulum of most hair follicles is then filled with comedone that consist of many accumulated layers of keratinized cells and sebum. Therefore, halogenated chemicals exhibit different actions, i.e. the induction of an immunologic consequence and the modulation of keratinocyte biology. These two conditions also provide good experimental models for investigating dermatology. PMID- 12850301 TI - Analysis of epidermal-type transglutaminase (transglutaminase 3) in human stratified epithelia and cultured keratinocytes using monoclonal antibodies. AB - BACKGROUND: Epidermal-type transglutaminase (TGase 3) is involved in the cross linking of structural proteins in the epidermis, which results in the formation of the cornified envelope. TGase 3 is activated by limited proteolysis of a 77 kDa zymogen during keratinocyte differentiation. OBJECTIVE: To characterize the expression of TGase 3 in human epidermis and cultured keratinocytes, we established specific monoclonal antibodies against the TGase 3. METHODS: Recombinant proteins for human TGase 3 produced in bacteria and baculovirus infected insect cells were purified as an antigen. Hybridomas are established and used for characterization of expression in epidermis and keratinocytes. RESULTS: Four antibodies were generated against recombinant human TGase 3, which reacted with the 77 kDa zymogen and in some cases either the 47 or 30 kDa active proteolytic fragments. In human epidermis and cultured keratinocytes, only the zymogen form of TGase 3 was detected. Immunohistochemical analysis of the skin revealed that the enzyme is present in the cells of the granular and cornified layers consistent with its role in cornified envelope formation. In cultured keratinocytes, TGase 3 was expressed in differentiating cells coincident with profilaggrin and keratin 10 expressions. CONCLUSION: Using monoclonal antibody against human TGase 3, we showed the expression of TGase 3 in upper layers of epidermis. TGase 3 displayed a diffuse cytoplasmic distribution in vitro consistent with its proposed role in the early phase of cornified cell envelope assembly in the cytoplasm. PMID- 12850302 TI - Evaluation of provocation test monitoring palmoplantar temperature with the use of thermography for diagnosis of focal tonsillar infection in palmoplantar pustulosis. AB - BACKGROUND: Since focal tonsillar infections are often associated with palmoplantar pustulosis (PPP), provocation tests have been performed for preoperative evaluation of tonsillectomy. However, these tests have not been fully established. OBJECTIVES: To introduce a more sensitive operative indication for tonsillectomy to the patients with PPP, we have monitored the temperature after provocation tests at palmoplantar sites, as measured by thermography, and we hypothesized that this methodology may lead to a more sensitive marker for tonsillectomy. METHODS: Twenty-two PPP patients with/without clinical tonsillitis were included in this study. After mechanical tonsillar massage, using infrared thermography, we have monitored the surface temperature at palmoplantar sites of 22 patients with PPP, five chronic tonsillitis patients without PPP, and four healthy controls, to compare the findings with the skin lesional outcome after tonsillectomy. RESULTS: There was a significant relationship between the effects of tonsillectomy and the results of provocation tests assessed by thermography. The sensitivity, specificity, and efficiency of the provocation tests with thermography of detecting a favorable outcome of tonsillectomy were 75.0, 83.3, and 77.3%, respectively, while those of the provocation tests as estimated with the conventional criteria were 37.5, 83.3, and 50.0%, respectively. CONCLUSION: Our results suggest that a new indicator using non-invasive thermography for the provocation tests is useful in predicting the effects of tonsillectomy for PPP. PMID- 12850303 TI - The effect of substance P on peripheral blood mononuclear cells in patients with atopic dermatitis. AB - BACKGROUND: There is increasing evidence that neuropeptides, especially substance P (SP), may be involved in the pathogenesis of atopic dermatitis (AD). OBJECTIVE: We performed this study to determine more precisely the role of SP in AD. METHODS: We separated peripheral blood mononuclear cells (PBMCs) from AD patients and normal controls, and measured proliferation response and cytokine release after adding SP (10(-11), 10(-10) and 10(-9) M). We also compared substance P receptor expression by semi-quantitative RT-PCR. RESULTS: PBMCs from AD patients proliferated at significantly higher rates (ca. by 30%). Semi-quantitative RT-PCR showed that the level of expression of SP receptor increased in AD patients versus normal controls. IL-4 release from PBMCs was significantly higher in AD patients, while IFN-gamma release from PBMCs was significantly lower in AD patients. Different concentrations of SP did not cause any difference in IL-4 and IFN-gamma secretions. However, TNF-alpha release from PBMCs in AD patients increased significantly at 10(-10) and 10(-9) M of SP compared to SP (-) control. IL-10 release from PBMCs increased significantly in AD patients with 10(-9) M of SP compared to SP (-) control. CONCLUSION: SP might aggravate AD by increasing the production of TNF-alpha and IL-10 rather than by affecting IL-4 and IFN gamma. This different immune response is considered to be the result of upregulated SP receptor in AD. PMID- 12850304 TI - Immunofluorescence studies using skin sections of recessive dystrophic epidermolysis bullosa patients indicated that the antigen of anti-p200 pemphigoid is not a fragment of type VII collagen. AB - BACKGROUND: There are a large number of autoimmune bullous diseases, which have distinct autoantibodies. Several reports on cases with IgG autoantibodies against a novel 200 kDa dermal protein have been published, for which we suggested the term, anti-p200 pemphigoid. However, the nature of this 200 kDa antigen has not been well characterized. OBJECTIVE: In this study, we examined the relationship between the 200 kDa protein and type VII collagen. METHODS: We collected sera from 12 cases of anti-p200 pemphigoid and skin sections from six cases of recessive dystrophic epidermolysis bullosa (RDEB). The reactivity of these sera was examined by indirect immunofluorescence using sections of the disease skin. RESULTS: we have shown that all the 12 anti-p200 pemphigoid sera could react with basement membrane zone of five cases of RDEB, while epidermolysis bullosa acquisita (EBA) sera were negative in these skins. In addition, in a case of RDEB, EBA sera reacted with intracytoplasmic deposition of type VII collagen, while no anti-p200 pemphigoid sera showed this reactivity. CONCLUSION: These results strongly suggested that the 200 kDa antigen is not a fragment of type VII collagen, but a specific autoantigen. PMID- 12850305 TI - Characterization of the human RAB38 and RAB7 genes: exclusion of new major pathological loci for Japanese OCA. AB - BACKGROUND: Oculocutaneous albinisms (OCAs) are due to various gene mutations that cause a disruption of melanogenesis in the melanocyte. Four different genes associated with human OCA have been reported, however, not all of OCA patients can be classified according to these four genes. We have sought to find a new major locus for Japanese OCA. Recently two genes, RAB38 and RAB7, were reported to play an important role in melanogenesis in the melanocyte, suggesting that these two genes could be good candidates for new OCA loci. OBJECTIVE: To determine the structures of the human RAB38 and RAB7 genes, and examine if the two genes are new major loci for Japanese OCA. METHODS: We screened mutations in these genes of 25 Japanese OCA patients who lacked mutations in the OCA1 and OCA2 genes with SSCP/heteroduplexes method. RESULTS: We determined the both genes, and their genomic organizations to design the primers for SSCP/heteroduplexes method. And then we screened mutations, but no mutation was detected. CONCLUSION: Neither of the genes is a new major locus for Japanese OCA. PMID- 12850306 TI - No involvement of IgG autoantibodies against extracellular domains of desmoglein 2 in paraneoplastic pemphigus or inflammatory bowel diseases. AB - BACKGROUND: Patients with paraneoplastic pemphigus (PNP) and inflammatory bowel diseases, such as Crohn's disease (CD) and ulcerative colitis (UC), develop autoantibodies against simple epithelial cells. About 20-30% of patients with PNP develop fatal bronchiolitis obliterans, in which autoantibody-mediated injury is suspected because of in vivo IgG deposition on cell surfaces of bronchial epithelia. OBJECTIVE: The purpose of this study is to determine whether patients with PNP, CD and UC have IgG autoantibodies against desmoglein 2 (Dsg2), which is expressed in all desmosome-bearing cells including respiratory and intestinal epithelia. METHODS: A secreted form of recombinant Dsg2 (rDsg2-His) which contains its entire extracellular domains was produced by baculovirus expression. The reactivity of patients' sera against rDsg2-His was examined by ELISA as well as immunoprecipitation. RESULTS: An anti-Dsg2 mouse monoclonal antibody, 6D8, showed positive reactivity against rDsg2-His in both methods. However, none of 38 PNP sera reacted with rDsg2-His by ELISA and none of 15 PNP sera tested immunoprecipitated rDsg2-His. Furthermore, none of 12 CD or 27 UC sera reacted with rDsg2-His by ELISA. CONCLUSION: These findings indicate that IgG autoantibodies against Dsg2 are not involved in PNP, CD or UC and suggest the existence of other unknown cell surface target antigen(s) in bronchiolitis obliterans in PNP. PMID- 12850307 TI - Identification of androgen-regulated genes in SV40-transformed human hair dermal papilla cells. AB - BACKGROUND: Androgens are major regulators of human hair growth, which exert their effect on follicular epithelium via the mesenchyme-derived dermal papilla (DP) cells. However, very few data are available with regard to the genes regulated by androgen in DP cells. OBJECTIVE: To investigate the differentially expressed genes by androgen in DP cells. METHODS: Human hair DP cells were cultured and transformed with SV40 T antigen. Using cDNA representational difference analysis, androgen-regulated genes in SV40-transformed dermal papilla (SDP) cells were identified. RESULTS: SDP cells revealed extended lifespan as compared with primary cultured DP cells. SDP cells expressed androgen receptor (AR) and showed androgen responsiveness. cDNA representational difference analysis followed by Northern blot analysis showed that heat shock cognate protein, Hsc70 was differentially expressed by dihydrotestosterone (DHT) treatment in SV40-transformed DP cells. CONCLUSION: These results suggest that Hsc70 may be involved in androgen action on DP cells. PMID- 12850308 TI - PCR-based identification of common dermatophyte species using primer sets specific for the DNA topoisomerase II genes. AB - BACKGROUND: We have determined nucleotide sequences of the DNA topoisomerase II genes of the dermatophyte species, and conducted a PCR-based identification system using species-specific primers for the nucleotide sequences. OBJECTIVE: To identify the major dermatophytes, Trichophyton rubrum, T. mentagrophytes, T. violaceum, M. gypseum, M. canis and E. floccosum, by PCR amplifications at the species level, without determining the nucleotide sequence. METHODS: For PCR based identification of the major dermatophyte species, a common primer set (dPsD1) for these species and species-specific primer sets (PsT and PsME) for each species were designed based on the genomic sequences of the DNA topoisomerase II genes of the dermatophytes, and tested for their specificities in PCR amplifications. The method consisted of amplification of the genomic DNA topoisomerase II gene by the common primer set, followed by a second PCR with the primer sets consisting of species-specific primers for each dermatophyte species. RESULTS: Using dPsD1, a DNA fragment of 3390 bp was amplified from the genomic DNA of all the dermatophyte species. In the subsequent nested PCR using species specific primer sets (PsT and PsME), both sets amplified unique sizes of PCR products, all of which corresponded to a species of the dermatophytes even in the presence of other fungal DNA. CONCLUSION: We demonstrate that the PCR-based identification targeting the DNA topoisomerase II gene is rapid and simple, and is available as a tool for the identification of the major dermatophyte species. PMID- 12850309 TI - Cytotoxic T lymphocyte antigen-4 gene (CTLA4) polymorphisms in Japanese patients with psoriasis vulgaris. PMID- 12850310 TI - Hypothesis: paradoxical absence of cellular immuno-deficiency in X-linked recessive ichthyosis and its explanation. PMID- 12850311 TI - A Bayesian neural network approach for modelling censored data with an application to prognosis after surgery for breast cancer. AB - A Bayesian framework is introduced to carry out Automatic Relevance Determination (ARD) in feedforward neural networks to model censored data. A procedure to identify and interpret the prognostic group allocation is also described. These methodologies are applied to 1616 records routinely collected at Christie Hospital, in a monthly cohort study with 5-year follow-up. Two cohort studies are presented, for low- and high-risk patients allocated by standard clinical staging. The results of contrasting the Partial Logistic Artificial Neural Network (PLANN)-ARD model with the proportional hazards model are that the two are consistent, but the neural network may be more specific in the allocation of patients into prognostic groups. With automatic model selection, the regularised neural network is more conservative than the default stepwise forward selection procedure implemented by SPSS with the Akaike Information Criterion. PMID- 12850312 TI - Active subgroup mining: a case study in coronary heart disease risk group detection. AB - This paper presents an approach to active mining of patient records aimed at discovering patient groups at high risk for coronary heart disease (CHD). The approach proposes active expert involvement in the following steps of the knowledge discovery process: data gathering, cleaning and transformation, subgroup discovery, statistical characterization of induced subgroups, their interpretation, and the evaluation of results. As in the discovery and characterization of risk subgroups, the main risk factors are made explicit, the proposed methodology has high potential for patient screening and early detection of patient groups at risk for CHD. PMID- 12850313 TI - Analyzing tumor gene expression profiles. AB - A brief introduction to high throughput technologies for measuring and analyzing gene expression is given. Various supervised and unsupervised data mining methods for analyzing the produced high-dimensional data are discussed. The main emphasis is on supervised machine learning methods for classification and prediction of tumor gene expression profiles. Furthermore, methods to rank the genes according to their importance for the classification are explored. The approaches are illustrated by exploratory studies using two examples of retrospective clinical data from routine tests; diagnostic prediction of small round blue cell tumors (SRBCT) of childhood and determining the estrogen receptor (ER) status of sporadic breast cancer. The classification performance is gauged using blind tests. These studies demonstrate the feasibility of machine learning-based molecular cancer classification. PMID- 12850314 TI - An Epicurean learning approach to gene-expression data classification. AB - We investigate the use of perceptrons for classification of microarray data where we use two datasets that were published in [Nat. Med. 7 (6) (2001) 673] and [Science 286 (1999) 531]. The classification problem studied by Khan et al. is related to the diagnosis of small round blue cell tumours (SRBCT) of childhood which are difficult to classify both clinically and via routine histology. Golub et al. study acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). We used a simulated annealing-based method in learning a system of perceptrons, each obtained by resampling of the training set. Our results are comparable to those of Khan et al. and Golub et al., indicating that there is a role for perceptrons in the classification of tumours based on gene-expression data. We also show that it is critical to perform feature selection in this type of models, i.e. we propose a method for identifying genes that might be significant for the particular tumour types. For SRBCTs, zero error on test data has been obtained for only 13 out of 2308 genes; for the ALL/AML problem, we have zero error for 9 out of 7129 genes that are used for the classification procedure. Furthermore, we provide evidence that Epicurean-style learning and simulated annealing-based search are both essential for obtaining the best classification results. PMID- 12850315 TI - Extracting multisource brain activity from a single electromagnetic channel. AB - This paper develops a methodology for the extraction of multisource brain activity using only single channel recordings of electromagnetic (EM) brain signals. Measured electroencephalogram (EEG) and magnetoencephalogram (MEG) signals are used to demonstrate the utility of the method on extracting multisource activity from a single channel recording. At the heart of the method is dynamical embedding (DE) where first an appropriate embedding matrix is constructed out of a series of delay vectors from the measured signal. The embedding matrix contains the information we require, but in a mixed form which therefore needs to be deconstructed. In particular, we demonstrate how one form of independent component analysis (ICA) performed on the embedding matrix can deconstruct the single channel recording into its underlying informative components. The components are treated as a convenient expansion basis and subjective methods are then used to identify components of interest relevant to the application. The framework has been applied to single channels of both EEG and MEG recordings and is shown to isolate multiple sources of activity which includes: (i) artifactual components such as ocular, electrocardiographic and electrode artefact, (ii) seizure components in epileptic EEG recordings, and (iii) theta band, tumour related, activity in MEG recordings. The results are intuitive and meaningful in a neurophysiological setting. PMID- 12850316 TI - Linking clinical data using XML topic maps. AB - Most clinical data is narrative text and often not accessible and searchable at the clinical workstation. We have therefore developed a search engine that allows indexing, searching and linking different kinds of data using web technologies. Text matching methods fail to represent implicit relationships between data, e.g. the relationship between HIV and AIDS. The international organization for standardization (ISO) topic maps standard provides a data model that allows representing arbitrary relationships between resources. Such relationships form the basis for a context sensitive search and accurate search results. The extensible markup language (XML) standards are used for the interchange of data relationships. The approach has been applied to medical classification systems and clinical practice guidelines. The search engine is compared to other XML retrieval methods and the prospect of a "semantic web" is discussed. PMID- 12850317 TI - Thermal hazard assessment of AN and AN-based explosives. AB - Ammonium nitrate (AN) is an essential ingredient in most fertilizers. It is also widely used in the commercial explosives industry. In this latter application, it is mostly mixed with fuel oil to form the most popular commercial explosive: ANFO. In both the fertilizer and the explosive industry, aqueous AN solutions (ANS) of various concentrations are processed. These solutions also form the basis of ammonium nitrate emulsion explosives (also called ammonium nitrate emulsions or ANE), which are produced either in bulk or in packaged form. For all these AN-based products, quantities of the order of 20,000kg are being manufactured, transported, stored, and processed at elevated temperatures and/or elevated pressures. Correspondingly, major accidents involving overheating of large quantities of these products have happened in several of these operations. In comparison, convenient laboratory quantities to investigate thermal decomposition properties are generally less than 1kg. As a result, in order to provide information applicable to real-life situations, any laboratory study must use techniques that minimize heat losses from the samples to their environment. In the present study, two laboratory-scale calorimeters providing an adiabatic environment were used: an accelerating rate calorimeter (ARC) and an adiabatic Dewar calorimeter (ADC). Experiments were performed on pure AN, ANFO, various ANS systems, and typical bulk and packaged ANE systems. The effects of sample mass, atmosphere, and formulation on the resulting onset temperatures were studied. A comparison of the results from the two techniques is provided and a proposed method to extrapolate these results to large-scale inventories is examined. PMID- 12850318 TI - Utilization of industrial waste products as adsorbents for the removal of dyes. AB - A number of low cost adsorbents from steel and fertilizer industries wastes have been prepared and investigated for the removal of anionic dyes such as ethyl orange, metanil yellow and acid blue 113 from aqueous solutions. The results indicate that inorganic wastes, i.e. blast furnace dust, sludge and slag from steel plants are not suitable for the removal of organic materials, whereas a carbonaceous adsorbent prepared from carbon slurry of fertilizer industry was found to adsorb 198, 211 and 219mg/g of ethyl orange, metanil yellow and acid blue 113, respectively. The adsorption of dyes on this adsorbent was studied as a function of contact time, concentration, particle size and temperature by batch method. The adsorption isotherm conformed to Langmuir model and the adsorption was found to be exothermic and physical in nature. Kinetic data conforms to Lagergren's equation with good correlation coefficients varying from 0.9998 to 0.9999 indicating that the adsorption is a first-order process. The adsorption data on carbonaceous adsorbent was compared to a standard activated charcoal sample and it was found that the prepared adsorbent is about 80% as efficient as standard activated charcoal and therefore, can be used as low cost alternative ( approximately 100 US dollars per ton) for colour removal from effluents. PMID- 12850319 TI - Modified dissolved organic matter fractionation technique for natural water. AB - A technique to fractionate dissolved organic materials (DOMs) from low DOM water (<5mg/l) was developed by using triple columns of DAX-8 adsorption resin, one column of AG-MP-50 cationic resin, and another column of WA 10 weak anionic resin in sequence. The procedure was then applied to fractionate water samples obtained at various sampling locations throughout two surface water treatment plants (WTPs) in central New Jersey to study its effectiveness, DOM occurrence, and variation along treatment units. The treatment plants utilize different treatment methods, hence producing variability in DOM fractions suitable for examining the procedure's effectiveness. This procedure was compared with current fractionation protocols and proved to be accurate in fractionation of low DOM water. PMID- 12850320 TI - Beneficial use of meat and bone meal combustion residue: "an efficient low cost material to remove lead from aqueous effluent". AB - Meat and bone meal (MBM) combustion residues, a natural apatite-rich substance, was evaluated as a low cost substitute for hydroxyapatite in lead sequestration from water effluents. The thermal behaviour of crude meat and bone meal was followed by TGA and 24% inorganic residue was collected. The resulting ashes were characterised by powder X-ray diffraction (XRD), particle size distribution, specific surface area (BET), and elemental analysis confirming apatite contents, with high level of phosphate (56.3%) and calcium (36.8%). Mechanism and kinetics of lead removal by this bioinorganic material were investigated and compared to mechanisms and kinetics involved with synthetic apatite. Batch metal removal experiments were carried out with 500 and 1500ppm (mg/kg) Pb(2+) solutions. Lead concentration, calcium and pH were monitored. We observed that the mechanism is similar to that occurring for pure apatite, and involved both surface complexation and calcium hydroyapatite (CaHA), Ca(10)(PO(4))(6)(OH)(2), dissolution followed by less soluble Pb(10)(PO(4))(6)(OH)(2) precipitation, as confirmed by XRD analysis of ashes after incubation with lead solution. Our results show that this natural apatite-rich material removes in a few minutes a large quantity of lead (275mg/g capacity) which remains however lower than the theoretical maximum capacity (if calcium were totally substituted by lead). Meat and bone meal combustion residues represent a valuable alternative apatite source for environmental application. PMID- 12850321 TI - Immobilization of mercury and zinc in an alkali-activated slag matrix. AB - The behavior of heavy metals mercury and zinc immobilized in an alkali-activated slag (AAS) matrix has been evaluated using physical property tests, pore structure analysis and XRD, TG-DTG, FTIR and TCLP analysis. Low concentrations (0.5%) of mercury and zinc ions had only a slight affect on compressive strength, pore structure and hydration of AAS matrixes. The addition of 2% Hg ions to the AAS matrix resulted in a reduction in early compressive strength but no negative effects were noticed after 28 days of hydration. Meanwhile, 2% Hg ions can be effectively immobilized in the AAS matrix with the leachate meeting the USEPA TCLP mercury limit. For a 2% Zn-doped AAS matrix, the hydration of the AAS paste was greatly retarded and the zinc concentration in the leachate from this matrix was higher than 5mg/l even at 28 days. Based on these results, we conclude that the physical encapsulation and chemical fixation mechanisms were likely to be responsible for the immobilization of Hg ions in the AAS matrix while only chemical fixation mechanisms were responsible for the immobilization of Zn ions in the AAS matrix. PMID- 12850324 TI - A tale of two necessities: breakaway technology versus diabetes. AB - Diabetes is endemic in industrialized society. A new technology is being used to uncover molecules that block a key enzyme in the diabetes disease mechanism. PMID- 12850325 TI - Targeting cancer: a NOvel way. AB - A nitric oxide (NO) prodrug has been shown to selectively kill NO-sensitive leukemia cells, leaving healthy cells unharmed. PMID- 12850326 TI - Nanodevices hold promise for gene therapy. AB - Nanodevices have been developed that are able to target, bind and cleave DNA. This novel approach could become the next big thing in gene therapy. PMID- 12850329 TI - Multisite clinical trials in strict accordance with GMP standards? AB - Thomas Poe looks at what the future holds for global research using today's technology and provides a clear roadmap showing how to get there efficiently. PMID- 12850330 TI - P53: an old dog begins to learn new tricks. PMID- 12850331 TI - Proteasome inhibitors in the treatment of cancer. PMID- 12850332 TI - Squaring the circle of high-throughput chemistry. AB - Andrew Lemon reviews the 225th national meeting of the American Chemical Society held in New Orleans, USA between the 23rd and 27th March 2003. PMID- 12850333 TI - High-content assays for ligand regulation of G-protein-coupled receptors. AB - High-content assays rely on the imaging of cellular events. They can be used to monitor the activation of G-protein-coupled receptors (or other receptors), their internalization into the cell, or alterations in their amount. In addition, multiplexed assays can provide further information about the characteristics of the receptor. Recent improvements in throughput using high-content screening platforms means that such assays are now an integral element of functional analysis in the drug discovery process. PMID- 12850334 TI - Continuous flow reactors for drug discovery. AB - To develop a new generation of drugs, pharmaceutical companies need to be able to synthesize and screen novel chemicals with enhanced speed. New technology that would enable a cost-neutral increase in the number of potential drug candidates would provide a distinct competitive advantage. The miniaturisation of chemical reactors offers many fundamental and practical advantages of relevance to the pharmaceutical industry, which is constantly searching for controllable, information-rich, high-throughput and environmentally friendly methods of producing compounds with a high degree of chemical selectivity. This article reviews the current and future applications of micro reactors that could enhance the drug discovery process. PMID- 12850335 TI - Chemical substructures in drug discovery. AB - The widespread use of HTS and combinatorial chemistry techniques has led to the generation of large amounts of pharmacological data, which, in turn, has catalyzed the development of computational methods designed to reduce the time and cost in identifying molecules suitable for pharmaceutical development. This review focuses on the use of substructure-based in silico techniques for lead discovery, an effective and increasingly popular approach for augmenting the chance of selecting drug-like compounds for preclinical and clinical development. PMID- 12850338 TI - How immune mechanisms are affected by pregnancy. AB - Pregnancy requires physiologic adaptations in all maternal systems, including the immune system. This process is complex and includes modifications at different levels and compartments of the maternal immune system. Although many of these changes are only partially explored and understood, recent investigations have showed that during pregnancy maternal circulating immune cells undergo modifications in cell counts, phenotypes, functions and ability to produce soluble factors, such as cytokines. The ultimate goal is to establish and maintain a successful pregnancy, which involves a state of selective immune tolerance, immunosuppression and immunomodulation in the presence of a strong anti-microbial immunity. The mammalian immune system has evolved to coexist with these needs by down-regulating potentially dangerous T-cell-mediated immune responses, while activating certain components of the innate immune system, such as monocytes and neutrophils. This unique dysregulation between different components of the immune system plays a central role in the maternal adaptation to pregnancy. PMID- 12850339 TI - Maternal immunization and malaria in pregnancy. AB - Malaria in pregnancy is a major cause of poor outcomes for both mother and fetus, and malaria-induced low birth weight (LBW) may kill nearly 400,000 African infants each year. The hallmark of pregnancy malaria due to Plasmodium falciparum is the sequestration of infected erythrocytes (IEs) in the placenta. Malaria can suppress responses to immunogens, and placental malaria can impair materno-fetal antibody transfer, potentially reducing the benefits of maternal immunization strategies. Parasites infecting the placenta have been shown to have distinct adhesive and antigenic features. Women become resistant to pregnancy malaria over successive pregnancies as they acquire antibodies that recognize placental parasites, suggesting that a vaccine is feasible. Serologic evidence indicates that the target for such a vaccine is antigenically conserved, and surface proteins expressed by placental parasites have been identified as vaccine candidates. Because malaria suppresses vaccine responses and reduces the transfer of maternal antibodies to the fetus, the prevention of pregnancy malaria may benefit maternal immunization strategies that protect neonates from infections other than malaria. PMID- 12850340 TI - Ex vivo human placenta models: transport of immunoglobulin G and its subclasses. AB - The comparison between fetal and maternal levels of immunoglobulins indicate that the human placenta during pregnancy develops a specific transport mechanism for IgG in the maternal to fetal direction. There are differences for the four subclasses, with preferential transfer of IgG(1) while the slowest transfer is seen for IgG(2). Under in vitro perfusion conditions of human term placenta IgG, when compared to other proteins, showed a significantly higher transfer rate of its subclasses from the maternal to the fetal side, indicating a specific transport mechanism. There is a preferential transfer rate, highest for IgG(1) and lowest for IgG(2), similar to those observed under in vivo conditions. The similarity in transfer of anti-TT-IgG and tetanus antigen, which was observed under in vivo and in vitro conditions, may suggest the transport as antibody antigen complex. PMID- 12850341 TI - Placental transport of immunoglobulin G. AB - Maternal antibodies transported across the placenta protect the newborn. Maternal immunoglobulin G (IgG) concentrations in fetal blood increase from early in the second trimester through term, most antibodies being acquired during the third trimester. IgG1 is the most efficiently transported subclass and IgG2 the least. Transfer across the syncytiotrophoblast of the chorionic villi is mediated by the neonatal Fc receptor, FcRn. Immune complexes are absorbed in the stroma of the villi, probably by FcgammaRI, FcgammaRII, and FcgammaRIII on placental macrophages. The mechanism of IgG transport across the endothelium of fetal capillaries is not understood. Endothelial cells in terminal villi express FcgammaRIIb. However, it is not known whether this receptor transports IgG or prevents transport of immune complexes to the fetus. PMID- 12850343 TI - Transfer of antibody via mother's milk. AB - Differing from humans, IgG from breast milk in many animal species (rodents, bovines, cats, ferrets, etc.) are transported across the intestinal epithelium into the neonatal circulation. This transport is located at the duodenal and jejunal level where enterocytes express a surface membrane receptor able to bind Fc of IgG and to facilitate transcytosis of these immunoglobulins. Fcgamma-R, which is very similar to the placenta receptor responsible for active transplacental transfer of IgG in humans, binds IgG but not other isotypes. Maternal milk antibodies represent an important part of circulating IgG in these animals, as they are involved in the negative feedback of endogenous IgG synthesis. This phenomenon stops abruptly as soon as weaning takes place. Neonatal calves that have a defect in such transfer of maternal immunoglobulins are at high risk of systemic infectious diseases. In humans, in whom gut closure occurs precociously, breast milk antibodies do not enter neonatal/infant circulation. A large part of immunoglobulins excreted in milk are IgA that protect mainly against enteric infections. The specificity of maternal milk IgA is driven by an entero-mammary cell circulation. Human milk also contains anti idiotypic antibodies capable of enhancing infant antibody response. Maternal milk antibodies coat infant mucosal surfaces and some have a clear protective role. This has been studied extensively in infectious disease models such as rotavirus, E. coli, poliovirus, and retroviruses. In the rotavirus model, antirotaviral IgA can be detected in stools of breast-fed but not bottle-fed neonates. In a large cohort of lactating women infected with HIV-1 in Rwanda, anti-HIV milk antibodies of the IgG isotype were more frequently detected followed by secretory IgM. Surprisingly, anti-HIV-1 SIgA were less frequently found. The presence of milk SIgA at 15 days as well as the persistence of a SIgM response during the whole lactation period was associated with lower risk of HIV transmission from the mother to the infant. Recently, HIV-1 antibodies from maternal milk have been shown to block transcytosis in vitro in a monolayer enterocyte model. Among these antibodies, those directed against the ELDKWA epitope had higher neutralising activity than serum antibodies. In humans, milk excreted antibodies play a major role in protecting infants from infection by pathogens having a mucosal portal of entry. PMID- 12850342 TI - Antibody protection: passive immunization of neonates against oral AIDS virus challenge. AB - We have established models for intrapartum and milk-borne HIV transmission by orally challenging neonatal macaques with chimeric simian-human immunodeficiency viruses (SHIVs). This allowed us to test safety and efficacy of passive immunization with human neutralizing monoclonal antibodies (nmAbs), which had been isolated from HIV clade B-infected individuals and which target conserved, functionally important epitopes. The nmAbs studied were F105 or IgG1b12, b12 for short (directed against the CD4 binding site), 2G12 (anti-gp120), 2F5 and 4E10 (both anti-gp41). Out of 16 newborn macaques challenged orally with different SHIV strains, 11 were completely protected by triple or quadruple nmAb combinations, even by post-exposure prophylaxis. In vitro, the combination of b12, 2G12, 2F5 and 4E10 potently neutralized primary HIV isolates of clades A, B, C, and D. Our data suggest that passive immunization with currently available anti-HIV clade B nmAbs could play a role in preventing transmission of non-clade B isolates through breastfeeding. We furthermore conclude that the epitopes recognized by the nmAbs in our successful passive immunization studies are important determinants for protection and provide targets for developing neutralizing antibody-response-based, active AIDS vaccines. PMID- 12850344 TI - The influence of mastitis on antibody transfer to infants through breast milk. AB - Clinical and subclinical mastitis are common among lactating women but their potential influence on programmes to improve infant health through maternal vaccination has not previously been addressed. Maternal antibodies, acquired either through natural infection or through vaccination, enter milk through two main pathways: by transcytosis or paracellularly. Transfer through both pathways is likely to be increased during mastitis, mediated by inflammatory cytokines. The increases are of similar order of magnitude to the variation in milk antibody concentrations between women within a population or between women in developing and developed countries. Thus, the effects of mastitis on vaccination programmes can be subsumed under the larger question of how breastfeeding in general affects these. PMID- 12850345 TI - Mucosal immunity: integration between mother and the breast-fed infant. AB - Lactating mammary glands are part of an integrated mucosal immune system with local production of antibodies, mainly consisting of secretory immunoglobulin A (SIgA). These antibodies generally reflect antigenic stimulation of mucosa associated lymphoid tissue (MALT) by common intestinal and respiratory pathogens. Antibodies in breast milk are thus highly targeted against infectious agents in the mother's environment, which are those likely to be encountered by the infant shortly after birth. Therefore, breast-feeding represents an ingenious immunological integration of mother and child. The mucosae are favored as portals of entry by most infectious agents, and the neonatal period is particularly critical in this respect. Mucosal pathogens are a major killer of children below the age of 5 years, being responsible for more than 14 million deaths annually. Diarrheal disease alone claims a toll of 5 million children per year in the developing countries. Epidemiological data suggest that the risk of dying from diarrhea could be reduced 14-24 times in breast-fed children. A beneficial clinical effect is also apparent in the industrialized world, even in relation to relatively common diseases such as otitis media and acute lower respiratory tract infections. PMID- 12850346 TI - Effect of maternal antibodies on the infant immune response. AB - Transplacentally-acquired antibodies are important for protection of infants in the early months of life. Maternal antibodies may dampen the infant's primary antibody response to infection or immunization. Even without a measurable response to the initial antigen exposure, the infant will usually have evidence of priming at the time of repeat exposure. No evidence for the development of fetal tolerance after immunization during pregnancy has occurred subsequently in infants following active vaccination. PMID- 12850347 TI - The influence of maternal immunization on light chain response to Haemophilus influenzae type b vaccine. AB - The repertoire of human antibodies to Haemophilus influenzae type b polysaccharide (Hib PS) has been extensively characterized. In response to Hib PS vaccines, both IgG1 and IgG2 antibodies are produced. Based on the Vkappa gene used for the antibodies, heamophilus antibodies can be divided into A2 and non-A2 antibodies. A2 antibodies account for the majority of heamophilus antibodies, are generally more functional, and are specific for heamophilus polysaccharide. In contrast, non-A2 antibodies cross-react with Escherichia coli K100. Studies of placental transfer showed that IgG1 antibodies cross the placenta better than IgG2 antibodies and the A2 antibodies better than non-A2 antibodies. Babies born to mothers who were immunized with a heamophilus vaccine had a high level of antibodies to Hib PS but the antibody level decreased during the first 6 months. The children also responded to a heamophilus conjugate vaccine. PMID- 12850348 TI - Measles and mumps vaccination as a model to investigate the developing immune system: passive and active immunity during the first year of life. AB - Evaluations of neutralizing antibody responses in 6-, 9- and 12-month-old infants given measles or mumps vaccine indicated that 6-month-old infants had diminished humoral immune responses associated with passive antibody effects, but also had an intrinsic deficiency in antiviral antibody production, which was independent of passive antibody effects. In contrast, lower neutralizing antibody titers in 9 month-olds were related only to passive antibody effects. Measles and mumps specific T-cell proliferation and interferon-gamma (IFNgamma) production were induced by vaccination at 6, 9 or 12 months, regardless of passive neutralizing antibodies or age. These observations suggest a need to refine concepts about passive antibody interference and primary vaccine failure, taking into account the sensitization of antiviral T-cells, which occurs in the presence of passive antibodies and is observed in infants who do not develop active humoral immunity. A second dose of measles vaccine given at 12-15 months enhanced antiviral T-cell responses to measles in infants who were vaccinated at 6 or 9 months, and produced higher seroconversion rates. Since T-cell immunity is elicited under the cover of passive antibodies, the youngest infants benefit from the synergistic protection mediated by maternal antibodies and their own capacity to develop sensitized antiviral T-cells, which prime for subsequent exposures to the viral antigens. Conceptually, maternal immunization approaches with vaccines that can be given to women of child-bearing age before pregnancy, or that are safe for administration during pregnancy, should enhance passive antibody protection. Rather than being detrimental to infant adaptive immune responses, maternal vaccination can be coupled effectively with vaccine regimens that elicit priming of antiviral immune responses in infants during the first year of life. PMID- 12850349 TI - Mechanisms by which maternal antibodies influence infant vaccine responses: review of hypotheses and definition of main determinants. AB - Several mechanisms have been suggested as mediating the inhibitory influence of maternal antibodies (MatAb) on infant responses. This inhibition is B cell determinant-specific, depends on the ratio between MatAb titers at the time of immunization and the dose of vaccine antigen, and leaves infant T cell responses largely unaffected. Neutralization of vaccine replication or FcgammaRIIB-mediated inhibitory signalling to infant B cells would not account for these characteristics. In contrast, determinant-specific masking of B cell epitopes and APC uptake of MatAb:vaccine antigen immune complexes, followed by antigen processing and presentation, explain the pattern of pre-clinical and clinical responses to infant vaccines. This allows the definition of the main determinants of the influence of MatAb on infant immunity. PMID- 12850350 TI - Immunization of pregnant women: reproductive, medical and societal risks. AB - Establishing successful vaccine programs for pregnant women would be the quintessence of preventive medicine when you realize the preventive potential for reproductive problems of many of the new and old vaccines. The development of vaccines to prevent maternal, fetal and newborn disease is actually in its infancy. The risks and benefits are discussed in detail as well as the most appropriate and inappropriate time to immunize women of reproductive age and pregnant women. The great majority of risks are theoretical, but the problem is that birth defects and other developmental and reproductive problems are in the group of "diseases of affliction" which means that there are tremendous emotional upheavals in families affected with serious reproductive problems. The failures in reproduction are so common that immunization of a pregnant women has potential deleterious consequences. A consortium of government, academia and industry must work together and an appeal to the more responsible members of the law profession to solve the problem of non-meritorious litigation has be in place before manufacturers of vaccines will be willing to initiate the development of new vaccines. The potential for reducing the incidence of birth defects, prematurity and neonatal infectious disease exists, but it will be difficult to initiate these programs because vaccine makers may be unwilling to assume an additional burden of negligence litigation. Certainly, it is clear that we could provide a safe vaccine for Group B streptococcus and infant botulism that would be of immediate benefit and the potential for reducing other diseases is realistic. This is not going to be an easy task. PMID- 12850351 TI - The impact of maternal infection or immunization on early-onset autoimmune diabetes. AB - Maternal immune responses exert short and long-term influences on the newborn immune system. The transfer of maternal antibodies helps the newborn controlling infectious agents that are prevalent in the population. Furthermore, recent studies indicate that the fetus is able to mount inflammatory and cytotoxic T lymphocyte responses in the course of maternal infections. Herein, we discuss the relevance of these observations to the possible relation between maternal infection or immunization and childhood-onset autoimmune diabetes. PMID- 12850352 TI - Materno-fetal transmission of autoimmune disease. PMID- 12850353 TI - Non-genetic inheritable potential of maternal antibodies. AB - Maternal antibodies (IgG and IgA) not only provide passive protection against microbial infections, but also exert a variety of equally important active, idiotypically-mediated immunoregulatory functions. Since the generation of maternal antibodies depends entirely on the stimulation of the mother's immune system by external mainly thymus-dependent antigens, with long-lived antigen independent plasma cells in the bone marrow, maternal antibodies represent the mother's collective ontogenetic immunological experience. Although their stimulatory potential in mice is restricted to the neonatal imprinting period, maternal antibodies exert a life-long determinative influence which is even dominant over seemingly genetic predispositions. Therefore, the functional impact of maternal IgG antibodies appears phenotypically as a non-genetic inheritance. PMID- 12850354 TI - Developmental factors associated with risk for atopic disease: implications for vaccine strategies in early childhood. AB - There is growing interest in the potential interactions between infant vaccination and risk for development of atopic disease. The aspect of this issue which has dominated this debate concerns suggestions that infant vaccination may stimulate allergic sensitisation. These suggestions derive from retrospective epidemiological analyses and will remain speculative unless they can be confirmed in prospective studies, particularly as conflicting findings have been reported. However, there is a potentially more important issue surfacing in this debate, which entails the converse situation, i.e. that genetic risk for atopy influences capacity to respond to vaccination during infancy. Support for the latter possibility comes from recent studies on the role of developmental factors which determine immune competence during infancy, and attendant risk for inflammatory and infectious diseases. The relevant findings are reviewed briefly below. PMID- 12850355 TI - Maternal immunisation in developing countries. AB - Maternal immunisation could help to prevent the 2-3 million neonatal and early infant deaths that occur in the developing world each year. Determining the causes of neonatal and early infant deaths in developing countries is difficult as most occur at home. However, it is likely that at least half are due to infections, several of which might be prevented by maternal immunisation. Even in poor countries with few health facilities, a high percentage of pregnant women attend an antenatal clinic at least once during pregnancy. Thus, an effective delivery system for maternal immunisation already exists and, because of the success of maternal tetanus immunisation, this approach to the prevention of serious illness or death in young infants is widely accepted by the general population. However, the high prevalence of HIV and malaria found in pregnant women in some parts of the developing world, especially sub-Saharan Africa, could have an effect on the efficacy of maternal immunisation as both of these infections adversely affect placental function. Nevertheless, the potential of maternal immunisation to prevent early infant deaths in developing countries needs to be fully explored. The incidence of pneumococcal infections is high in many developing countries and about 25% of these infections occur at an age before protection could be anticipated following vaccination with a pneumococcal conjugate vaccine in infancy. Thus, a strong case can be made for a trial of the effectiveness of maternal immunisation with a pneumococcal vaccine in preventing serious illness or death in young infants in developing countries. PMID- 12850356 TI - Tetanus in developing countries: an update on the Maternal and Neonatal Tetanus Elimination Initiative. AB - Tetanus is a vaccine-preventable disease that yearly causes a total of 309,000 deaths. Of particular concern is maternal and neonatal tetanus (MNT), which can be prevented through immunization of the mother in pregnancy. In 2000, neonatal tetanus alone was responsible for an estimated 200,000 deaths. While the focus is on 57 priority countries, 90% of the neonatal tetanus deaths occur in 27 countries. UNICEF is spearheading the effort to eliminate MNT by the year 2005, with the support of numerous partners. MNT elimination is defined as less than one case of neonatal tetanus per 1000 live births at district level. The main strategies consist of promotion of clean delivery practices, immunization of women with a tetanus toxoid (TT) containing vaccine, and surveillance. Maternal tetanus immunization is, in most developing countries, implemented as part of the routine immunization program. However, large areas remain underserved, due to logistical, cultural, economical or other reasons. In order to achieve the target of MNT elimination by 2005, and to offer protection to women and children otherwise deprived from regular immunization services, countries are encouraged to adopt the "high risk approach". This approach implies that, in addition to routine immunization of pregnant women, all women of child bearing age living in high risk areas are targeted for immunization with three doses of a tetanus toxoid containing vaccine (TT or Td), implemented as "supplemental immunization activities" (SIAs). Through SIAs, about 17 million women have been reached with at least two doses of TT vaccine in the past 3 years, and it is estimated that another 200 million need to be targeted in the years to come. SIAs should substantially reduce the burden of disease. Countries will also have to improve their existing immunization and clean delivery programs to ensure that the elimination of maternal and neonatal tetanus is maintained. PMID- 12850357 TI - Studies of maternal immunisation with pneumococcal polysaccharide vaccine in Papua New Guinea. AB - In two studies, pneumococcal polysaccharide (Pnc PS) vaccine was given to more than 400 pregnant Papua New Guinean women. No deleterious effects were found. The vaccine prevented acute lower respiratory infection (ALRI) among offspring in utero or aged 1-17 months at the time of maternal immunisation, suggesting protection through breast feeding. Serum IgG antibody titres were higher in vaccinated than unvaccinated groups for 2-4 months after delivery and no immune suppression, evaluated by the response to subsequent Pnc PS vaccination, was detected. Breast milk IgA to four serotypes was 1.1-1.8 times higher in immunised than unimmunised women for 6 months postpartum. Given results from several developing countries, large-scale safety and efficacy trials are now justified. Postpartum maternal immunisation is another intervention under consideration. PMID- 12850358 TI - Maternal immunization with pneumococcal polysaccharide vaccine in the Philippines. AB - A randomized, controlled study was conducted to evaluate the immunogenicity and reactogenicity of the 23-valent pneumococcal (Pnc) polysaccharide (PS) vaccine among pregnant women and to ascertain the transfer of anti-Pnc antibody (Ab) from mother to infant. One hundred and sixty women received either one dose of Pnc PS vaccine, Haemophilus influenzae type b conjugate vaccine and tetanus toxoid (TT) (Pnc vaccine group, N=106) or TT only (control group, N=54). Sera were obtained from all mothers prior to vaccination and 4 weeks after from the vaccinated group. Cord blood was obtained in 75% of deliveries. Anti-Pnc Ab for serotypes 1, 5, 6B, 14, 18C and 19F was determined using enzyme immunoassay. The Pnc vaccine and control groups were comparable in terms of age, parity, gravidity, prior doses of TT, and pre-vaccination geometric mean concentration (GMC in microg/ml) of anti-Pnc Ab. Between 66 and 87% of the mothers had type-specific Ab prior to vaccination. There was a significant rise in anti-Pnc Ab (varying from 3.3- to 9.1-fold for the individual serotypes) between the pre and post-vaccination samples. Adverse reactions were mild and required no treatment. The level of anti Pnc Ab in cord blood was significantly lower in the control group compared to the Pnc vaccine group. Vaccination of pregnant women with Pnc Ps vaccine induces good immune response and Ab can be transferred to their infants via cord blood thus providing enhanced protection. PMID- 12850359 TI - Maternal immunization with Haemophilus influenzae type b vaccines in different populations. AB - Haemophilus influenzae type b (Hib) vaccines provide an excellent model for maternal immunization because effective vaccines are readily available and the vaccines are safe and reliable, and markers of efficacy have been established and standardized. Studies of polysaccharide and conjugate Hib vaccines administered to pregnant women and women of childbearing ages are reviewed in this paper. The type of vaccine has been shown to be important in increasing transplacental passage of maternal antibody. The timing of vaccine during pregnancy is also important in the transfer of this antibody. The total amount of IgG antibody in the mother, as well as the isotype class and subclass of IgG antibody, influences the final levels of antibody in the neonate. Placental integrity has been shown to be important in the active transport of antibody from mother to fetus. The impact of increased levels of Hib antibody in infants at the time of primary immunization with Hib does not appear to interfere with vaccine efficacy, although higher antibody levels in infants at the time of immunization may result in lower total antibody levels following all doses of vaccine. Principles observed in these studies have potential application against other important neonatal pathogens. PMID- 12850360 TI - Maternal immunization with inactivated influenza vaccine: rationale and experience. AB - Inactivated influenza vaccine is recommended for routine use in high-risk individuals in the United States, including women who will be in the second or third trimesters of pregnancy. The basis for this recommendation is the high risk of exposure and disease due to influenza viruses in pregnant women, as well as the impact of influenza virus infection on the fetus and infant. Historical data from the influenza pandemics of 1918 and 1957 illustrate the potential risks of this infection in pregnant women and their fetuses. Prospective studies have demonstrated higher cord antibody levels to influenza in babies born to mothers immunized during pregnancy, and a delay in the onset and decrease in severity of babies born with higher antibody levels. Increased influenza vaccine use during pregnancy has the potential to benefit both the woman and her infant. PMID- 12850361 TI - Safety and immunogenicity of respiratory syncytial virus purified fusion protein 2 vaccine in pregnant women. AB - A randomized, double-blind, placebo controlled study was carried out to determine the safety and immunogenicity of RSV PFP-2 vaccine (Wyeth-Lederle Vaccines, NY) in 35 healthy women in the third trimester of pregnancy and their offspring. Infants were followed during their first RSV season for occurrence and severity of respiratory illnesses. RSV-PFP-2 vaccine was safe and well tolerated by pregnant women. Mild pain at the site of injection occurred in 65% of PFP-2 and 13% of placebo recipients (P=0.005). There were no systemic reactions, fever, or serious adverse events associated with vaccine administration in mothers. All 35 infants were born healthy, and there were no differences among the groups in perinatal or neonatal outcomes, growth and development in the first year of life. During the RSV season, there was no increase in the frequency or morbidity associated with respiratory tract illnesses in infants of vaccine recipients. 15/20 (75%) vaccine recipients had a response to PFP-2 by Western blot vs. 0/15 placebo recipients (P<0.01). 19/20 (95%) vaccine recipients had a > or =4 fold rise in IgG ELISA Ab after immunization with PFP-2 vs. 0/15 placebo recipients (P<0.01). Geometric mean concentrations of IgG ELISA Ab were 4 fold higher in infants of vaccine recipients at birth, 2 and 6 months after delivery than in infants of placebo recipients (P<0.01). A modest (0.5log2) increase in neutralization Ab was observed in vaccine recipients and their infants. The half life of maternal antibodies in infants was > or =3 weeks. There was no evidence of enhanced T-cell or cytokine activity in infants of vaccine recipients vs. infants of placebo recipients. Vaccine specific anti-F IgA and IgG concentrations in breast milk were higher in mothers who received RSV-PFP-2. PMID- 12850362 TI - Immunization of pregnant women with group B streptococcal type III capsular polysaccharide-tetanus toxoid conjugate vaccine. AB - AIM: To determine the safety and immunogenicity of group B streptococcal (GBS) type III CPS-TT conjugate vaccine in pregnant women. METHODS: Prospective, randomized (2:1, vaccine:placebo), double-blind, placebo-controlled trial in women at 30-32 weeks' gestation and their infants. Immune responses were measured with IgG-specific ELISA assays; killing of III GBS by 1- and 2-month infant sera was assessed by opsonophagocytosis. RESULTS: Immunization was well-tolerated; maternal and infant outcomes were similar in vaccine and placebo groups. III-TT conjugate elicited good levels of IgG to III CPS that were functionally active against type III GBS through age 2 months. CONCLUSION: Maternal immunization with GBS CPS-TT conjugates could prevent maternal, neonatal and young infant GBS disease. PMID- 12850363 TI - Pneumococcal conjugate vaccines as maternal and infant immunogens: challenges of maternal recruitment. AB - Maternal pneumococcal immunization may be a strategy for the prevention of pneumococcal disease during the first months of life before infant-administered pneumococcal conjugate vaccine (PCV) becomes protective. Preparatory to a Phase III efficacy trial to investigate this strategy for preventing early infant otitis media (OM), we are conducting a Phase I-II randomized, double-masked trial to determine if maternal 9-valent PCV immunization alters active antibody production in infants who are immunized with PCV-7 and to compare local and systemic adverse events among women immunized with PCV-9 or placebo. A pilot study was conducted in 1997 in a Minneapolis-St. Paul health maintenance organization, HealthPartners (HP), to estimate enrollment rate for the proposed Phase III trial. A total of 154 pregnant women were selected of whom 122 appeared to be eligible from ICD codes in the HP claims database; 76 responded, and 46 of these were eligible for participation. Four of the 46 reported that they would be very likely and 21 somewhat likely to participate; an estimated 20% (25/122) participation rate. The records of 4,289 women in their second trimester have been reviewed to date for the Phase I/II trial; 2,589 (60%) were found not to be eligible; and an additional 1,015 (24%) could not be contacted. Of the 685 remaining, 558 (81%) were not interested in participating when interviewed, and 127 (19%) have consented. Most (46%) of those who declined participation have cited concern about the safety of vaccination during pregnancy. While the overall participation rate of second trimester women is only 3% (127/4,289), maternal attrition (2.8%) and infant attrition from the study before age 7 months (3.9%) are lower than projected (25%). Differences in study design could explain the difference in participation rates estimated in the pilot study and experienced in the Phase I/II study. Reports of safety from large-scale studies of maternal PCV immunization in industrialized countries are needed before women are receptive to maternal immunization to protect their infants against pneumococcal disease. PMID- 12850364 TI - Correlates of immunity to respiratory syncytial virus (RSV) associated hospitalization: establishment of minimum protective threshold levels of serum neutralizing antibodies. AB - OBJECTIVE: To determine if respiratory syncytial virus (RSV) specific, serum antibody titers correlate with protection against RSV associated-hospitalization at all ages. DESIGN: Participants who were enrolled in a trial to determine the frequency of specific virus infections associated with hospitalization [J. Am. Med. Assoc. 283 (2000) 499] were included in our analysis if they were enrolled from July 1991 to June 1993, had a culture for virus isolation, and provided blood samples at hospitalization and 14-60 days later. RSV infection was defined by a positive culture and/or serology. Microneutralization, ELISA to the fusion (F) protein and Western blot were the serological assays that were used to determine correlates of immunity. RESULTS: One hundred and seventy-five individuals, 1 month to 89 years old, out of 538 patients hospitalized with an acute respiratory infection met the criteria for analysis. RSV associated hospitalization occurred in 11 (40.7%) of 27 infants (<1 year), 8 (38.1%) of 21 young children (1 to <5 years), and 15 (11.8%) of 127 children and adults (> or =5 years). At the time of hospitalization, geometric mean neutralizing antibody titers (log(2)) to RSV/A and RSV/B, and geometric mean binding antibody titer (log(2)) to F protein were significantly higher in patients with non-RSV associated-hospitalization compared to those with RSV associated-hospitalization (RSV/A: 7.9 versus 6.1, P<0.001; RSV/B: 9.4 versus 7.3, P<0.001; ELISA-F, 13.9 versus 12.6, P=0.01). For every 1 log(2) increase in titer of neutralizing antibodies to RSV/A and RSV/B, and binding antibody to F protein there was a significant increase in the likelihood of not having an RSV associated hospitalization by 22.3, 25, and 24.4% respectively. A minimal protective threshold titer of > or =6.0 (odds ratio 3.5; 95% CI 1.4-9.1) and > or =8.0 log(2) (odds ratio 2.9; 95% CI 1.1-7.7) against RSV associated-hospitalization was established for neutralizing antibodies to RSV/A and RSV/B; a threshold titer could not be established for binding antibody to F protein. CONCLUSION: Participants with naturally acquired serum neutralizing antibody levels at least equal to the minimal protective threshold titer were approximately three times more likely not to have an RSV associated-hospitalization. We speculate that achieving a minimal protective threshold antibody titer through active immunization will significantly reduce RSV associated-hospitalization among all ages. PMID- 12850365 TI - Pertussis: an important target for maternal immunization. AB - Cord blood antipertussis IgG concentrations were measured in infants and were found to be nearly equal to maternal levels. By 4 months of age most infants had no measurable antibody to pertussis toxin (PT) or filamentous hemagglutinin. Higher concentrations of maternally derived antibody to PT were associated with a weaker pertussis toxin antibody response to whole cell pertussis vaccine, but not to a cellular vaccine. These studies suggest that maternal immunization would provide early protection of the newborn to allow time for the primary immunization schedule at 2, 4, and 6 months of age to induce more durable protection. PMID- 12850366 TI - Maternal immunization: US FDA regulatory considerations. AB - Vaccination of pregnant women provides important health benefits to both, mother and infant, and has been an important disease prevention strategy in these two groups. While most vaccines currently licensed in the US are not indicated for use during pregnancy, depending on the vaccine, vaccination programs do frequently include pregnant women. In addition, recent emphasis has been placed on maternal immunization strategies to protect young infants from severe infections. Currently, unless the vaccine is specifically indicated four maternal immunization, no data are collected regarding the vaccine's safety in pregnant women prior to licensure. However, more females of childbearing age participate in clinical trials and a broad range of novel vaccine products are in development indicated for adolescents and adults. Thus, there is increasing concern for the unintentional exposure of an embryo/fetus before information is available regarding the potential risk versus benefit of the vaccine. Since pregnant women are usually excluded from participation in clinical trials, conclusions regarding developmental risk at the time of licensure are frequently based solely on data derived from developmental toxicity studies in animal models. This paper will review regulatory, preclinical and clinical issues as they pertain to development programs for vaccines intended for vaccination during pregnancy. PMID- 12850367 TI - Vaccines in Argentina: a regulatory view. AB - In Argentina, vaccines for immuno-preventable diseases are regulated by the national regulatory agency, the Administracion Nacional de Medicamentos, Alimentos y Tecnologia Medica (the National Administration of Drugs, Food and Medical Devices, or ANMAT) created in 1992 to ensure efficacy and safety of drugs, food and medical devices available in the country, according to Law 16,463 and Decree 150/92. ANMAT has licensed 84 out of 157 vaccines registered in Argentina. Since 1994, ANMAT evaluated, approved and inspected 20 clinical trials with vaccines (1.8% of the 1062 trials approved by the agency since that time). The National System of Pharmaco-vigilance has received 318 communications of eventual adverse post-vaccination events (0.3% of the total). In addition, ANMAT provides support to the National Immunisation Programme. The current procedure is to follow international guidelines in the field, to be prepared for new, rapidly changing scenarios. PMID- 12850368 TI - Acceptability of tetanus toxoid vaccine by pregnant women in two health centres in Abidjan (Ivory Coast). AB - A study was conducted in two health centres in Abidjan, Ivory Coast (Abobo and Port Bouet) to compare the knowledge of pregnant women regarding tetanus and hepatitis B and to evaluate the acceptability of tetanus immunisation. A total of 124 women were interviewed. In spite of Information Education Communication (IEC) meetings held by midwives focusing on both diseases, knowledge about tetanus appeared to be substantially higher than that about hepatitis B. The acceptability of tetanus toxoid vaccine was good, the only barrier being the fear of useless injections. However, the risk of tetanus transmission during delivery should be better explained to women, who often give birth at home in the presence of traditional birth assistants. The awareness of men should also be raised by information campaigns, using different local networks. PMID- 12850369 TI - Session IV. Ethical, liability and regulatory issues. PMID- 12850371 TI - Cytogenetic study of malignant triton tumor: a case report. AB - Malignant triton tumor (MTT) is a highly malignant neoplasm, classified as a variant of malignant peripheral nerve sheath tumor (MPNST) with rhabdomyoblastic differentiation. Few cytogenetic studies of MTT have been reported using conventional cytogenetic analysis. Here, we report a comprehensive cytogenetic study of a case of MTT using G-banding, Spectral Karyotyping(), and fluorescence in situ hybridization (FISH) for specific regions. A complex hyperdiploid karyotype with multiple unbalanced translocations was observed: 48 approximately 55,XY,der(7)add(7)(p?)dup(7)[2],der(7) t(7;20)(p22;?)ins(20;19)[5],der(7)ins(8;7)(?;p22q36)t(3;8)t(8;20)[15],-8[5], 8[19],r(8)dup(8), +der(8)r(8;22)[4], 9[9],der(11)t(11;20)(p15;?)ins(20;19)[22],der(12)t(8;12)(q21;p13)[21],der(13) t(3;13)(q25;p11),-17,-19,der(19)t(17;19)(q11.2;q13.1),-20,-22,+4 approximately 7r[cp24]/46,XY[13]. The 1995 International System for Human Cytogenetic Nomenclature was followed where possible. Note that breakpoints were frequently omitted where only SKY information was known for a small part of an involved chromosome. Our analysis revealed some breakpoints in common with those previously reported in MTT, MPNST, and rhabdomyosarcoma, namely 7p22, 7q36, 11p15, 12p13, 13p11.2, 17q11.2, and 19q13.1. FISH showed high increase of copy number for MYC and loss of a single copy for TP53. PMID- 12850370 TI - Chromosomal instability in osteosarcoma and its association with centrosome abnormalities. AB - The mechanism that generates the extreme aneuploidy that characterizes osteosarcoma (OS) is poorly understood. In this study, interphase fluorescence in situ hybridization (FISH) analysis was used to enumerate cell-to-cell variation of several different chromosomes. We also investigated whether there was an association between TP53 mutation and centrosome aberrations in the generation of chromosomal aneuploidy in OS in four OS cell lines (HOS, SAOS2, U2OS, and MG63) and in a subset of seven tumors. Our analysis showed that there was a wide range of numerical changes affecting multiple chromosomes in OS cell lines and tumors. These data suggest that chromosomal instability (CIN) could be responsible for the extensive aneuploidy associated with this tumor. The results also showed an increased frequency of atypical mitotic figures in three OS cell lines with defective TP53, function and significantly, a more marked CIN phenotype was present in these lines. Furthermore, numerical aberrations of centrosomes were also present in these three OS cell lines with TP53 mutations. In two of three OS patients' tumors there was a large increase in the percentage of abnormal centrosome numbers. We conclude that CIN is a consistent feature of OS and that an intrinsic disturbance of the chromosomal segregation mechanisms is likely associated with centrosome aberrations. PMID- 12850372 TI - Localization of transfected B7-1 (CD80) DNA in human melanoma cells after particle-mediated gene transfer. AB - The purpose of this study was to evaluate stable DNA transfection of M-21 human melanoma cells with particle-mediated gene transfer (PMGT) with B7-1 cDNA and to identify sites of gene integration. Stable B7-1 transfectants (M-21-B7) were obtained with PMGT using a plasmid vector containing cDNA for both B7-1 and neomycin phosphotransferase, with subsequent selection with G418. The transfected cells were flow sorted by B7-1 expression into two populations, bright and dim. The bright population had 85%-90% of cells expressing B7-1; the dim population had less than 50% of cells with B7-1 expression. Chromosome analysis with fluorescence in situ hybridization (FISH) and G-banding showed that 70% of bright cells had two main integration sites, with extensive amplification of the transgene. The dim population had random signal distribution, with little or no amplification, despite G418 selection. Because B7-1 has been mapped to 3q21, FISH was performed using a chromosome 3 painting probe (WCP) together with a probe for B7-1. In transfected bright M-21 cells, amplified genes that hybridized with the B7-1 construct were localized to chromosome 3 material inserted into marker chromosomes. These data suggest that B7-1 insertion may involve homologous recombination, but maintenance of integration and amplification required selection. PMID- 12850373 TI - Recurrent genetic alterations in 26 colorectal carcinomas and 21 adenomas from Chinese patients. AB - Colorectal cancer (CRC) is one of the most common malignancies worldwide. The incidence of CRC in the Chinese population has increased dramatically during the last two decades; however, nonrandom chromosomal alterations in Chinese patients have not been described. In the present study, comparative genomic hybridization (CGH) was applied to detect recurrent chromosome alterations in 26 primary colorectal carcinomas and 21 colorectal adenomas from Chinese patients. In CRC, several recurrent chromosomal changes were found, including gains of 8q (14/26 cases, 54%), 20q (54%), 3q (50%), 13q (50%), 5p (46%), 7p (42%), 7q (42%), and 12p (38%) and losses of 18q (65%) and 17p (42%). From comparison with previous CGH studies, the frequent gains of 3q and 12p might be distinctive occurrences in Chinese patients. The distribution of frequently found chromosomal alterations in different locations was studied. The gain of 20q was more frequently found in colon cancer (P<0.01) and the gain of 12p was more frequently found in rectal cancer. Chromosomal alterations were found in 19/21 of adenomas; the most frequent chromosomal alteration was the loss of 18q (9/21 cases, 43%). These recurrent alterations provide several starting points for the isolation of candidate oncogenes and tumor suppressor genes. PMID- 12850374 TI - Cytogenetic and molecular genetic analyses of endometrial stromal sarcoma: nonrandom involvement of chromosome arms 6p and 7p and confirmation of JAZF1/JJAZ1 gene fusion in t(7;17). AB - Endometrial stromal sarcomas (ESS) are rare neoplasms with the capacity both to invade the myometrium locally and to give rise to extrauterine metastases. Cytogenetic abnormalities have been reported in 22 cases of ESS, mostly involving rearrangements of chromosomes 6, 7, and 17. The most characteristic translocation of this tumor type, t(7;17)(p15 approximately p21;q12 approximately q21), was recently shown to generate a JAZF1/JJAZ1 fusion gene. We report three additional cases of ESS with abnormal karyotypes, whose interpretation was based on the combined analysis by conventional cytogenetics and cross-species color banding FISH (RxFISH). The combination of G-banding and RxFISH in every case gave additional information beyond that obtained by either technique alone, determining the identity of even complex inter- as well as intrachromosomal rearrangements. In one of the three tumors, a t(7;17) was seen; molecular genetic studies identified the JAZF1/JJAZ1 fusion gene in this case. Two tumors had aberrations that included structural changes of chromosome arms 6p and 7p. Evidently, karyotypic, and hence pathogenetic, heterogeneity exists for tumors classified as endometrial stromal sarcomas based on their phenotypic features. PMID- 12850375 TI - Cytogenetic and molecular findings related to rhabdomyosarcoma. An analysis of seven cases. AB - Rhabdomyosarcoma (RMS) is the most common soft-tissue sarcoma in childhood. Histologically, it is subdivided histologically into two main subtypes: alveolar (ARMS) and embryonal (ERMS). ARMS is characterized by t(2;13)(q35;q14) or its variant t(1;13)(p36;q14), which fuse PAX3 and PAX7, respectively, with FKHR to produce chimeric genes. ERMS is frequently associated with loss of heterozygosity of 11p15.5. We investigated seven RMS (three ARMS and four ERMS) by means of cytogenetic, fluorescence in situ hybridization, and molecular analyses, including the study of the main genes implicated in the G1- to S-phase cell cycle transition, and correlated these studies with pathologic findings and clinical outcome. All tumors showed clonal, numerical, and structural chromosomal abnormalities. Two ARMS had the t(2;13)(q35;q14) and the third a PAX7/FKHR fusion, a cryptic t(1;13)(p36;q14), undetected by cytogenetic techniques, but revealed by reverse transcriptase polymerase chain reaction. One ERMS showed a der(11)t(3;11)(p21;p15) as a sole structural anomaly. Gene amplification was seen in four tumors, as double minutes or in the form of homogeneously staining regions. Overexpression of MYCN oncogene was found in two ARMS; N-myc DNA probe detected oncogene amplification located on the double minutes of these cases. Analysis of the regulatory genes responsible for G1- to S-phase transition showed a homozygous deletion of the 9p21 locus genes in a spindle-cell ERMS. PMID- 12850376 TI - Aberrant promoter methylation of multiple genes in oligodendrogliomas and ependymomas. AB - Promoter hypermethylation represents a primary mechanism in the inactivation of tumor suppressor genes during tumorigenesis. To determine the frequency and timing of hypermethylation during carcinogenesis of nonastrocytic tumors, we analyzed promoter methylation status of 10 tumor-associated genes in a series of 41 oligodendrogliomas (22 World Health Organization [WHO] grade II; 13 WHO grade III; 6 WHO grade II-III oligoastrocytomas) and 7 WHO grade II-III ependymomas, as well as 2 nonneoplastic brain samples, by a methylation-specific polymerase chain reaction. Aberrant CpG island methylation was detected in 9 of 10 genes analyzed, and all but one sample displayed anomalies in at least one gene. The frequencies of hypermethylation for the 10 genes were as follows, in oligodendrogliomas and ependymomas, respectively: 80% and 28% for MGMT; 70% and 28% for GSTP1; 66% and 57% for DAPK; 44% and 28% for TP14(ARF); 39% and 0% for THBS1; 24% and 28% for TIMP3; 24% and 14% for TP73; 22% and 0% for TP16(INK4A); 3% and 14% for RB1; and 0% in both neoplasms for TP53. No methylation of these genes was detected in normal brain tissue samples. We conclude that a high frequency of aberrant methylation of the 5' CpG island of the MGMT, GSTP1, TP14(ARF), THBS1, TIMP3, and TP73 genes is observed in nonastrocytic neoplasms. This aberration seems to occur early in the carcinogenesis process (it is already present in the low-grade forms), although in some instances (DAPK, THBS1, and TP73) it appears also associated with the genesis of anaplastic forms. PMID- 12850377 TI - A fluorescence in situ hybridization study of TEL-AML1 fusion gene in B-cell acute lymphoblastic leukemia (1984-2001). AB - Acute lymphoblastic leukemia (ALL) is associated with recurrent chromosomal abnormalities. The cryptic translocation t(12;21)(p13;q22), which leads to the TEL-AML1 fusion gene, is the most common abnormality in childhood B-cell ALL. The aim of this retrospective study was to determine the incidence of TEL-AML1 fusion in childhood and adult B-cell ALL using interphase fluorescence in situ hybridization (I-FISH) and its association with additional changes. FISH, using dual-color extra-signal (ES) DNA probe specific for the TEL and AML1 genes, was performed either on blast cells suspensions stored in liquid nitrogen immediately after Ficoll or on leukemic cells preserved in fixative solution at -20 degrees C after short-term culture. No TEL-AML1 fusion was observed in the 26 ALL adults. The fusion was found among 19.6% of the 57 ALL children, additional changes being identified by conventional cytogenetics in 80% of the cases. A deletion of the untranslocated TEL was observed in 36.3% of the ALL with the TEL-AML1 fusion. The coexpression of myelocytic and B-lymphoid antigens was found in 3 of the 11 of TEL-AML1 fusion positive-ALL. Our results (frequency of TEL-AML1 fusion in children and of the deletion of the untranslocated TEL allele, mean age of the patients and white blood cell count) are within the range observed by others. Structural chromosomal abnormalities other than the t(12;21) are frequent and may play a role in the prognosis of these patients. PMID- 12850378 TI - Chromosomal aberrations in premalignant and malignant squamous epithelium. AB - Biopsies of oropharyngeal cancer were screened for chromosomal imbalances by comparative genomic hybridization (CGH) performed on 22 primary tumors and morphologically nonmalignant surrounding mucosa. The aim was to determine early chromosomal changes of tumor development and to draw conclusions on the mechanisms leading to multiple tumors. The most prominent chromosomal imbalances observed were over representations of genomic material on 3q, 15q, 8q, and 11q and losses on 9p, 3p, and 11q. In morphologically normal mucosa collected at 1 cm from the primary tumor border (M1), amplifications on 15q and 21q were most frequent. Far fewer gains and losses were found in M1 than in the primary tumor (average 2.2 vs. 6.9). Gains dominated over losses, but a tendency toward an increasing proportion of losses in the primary tumor (PT) was observed (ratio of gains to losses: PT, 4.75; M1, 6.3). Almost all the imbalances in M1 were detected in the primary tumor. No chromosomal alterations were identified with CGH in tissue samples dissected at 2 cm from the primary tumor (M2). In all samples, dysplastic morphologic changes decreased with distance from the primary tumor, which correlates with the observed lower level of genetic changes. We suggest that gains of genetic material on 15q and 21q are early events in malignant progression of squamous cell carcinoma, followed by gains on 3q, 8q, and 11q, and losses on 3p and 9p at later stages. Based on our cytogenetic data, we discuss the monoclonal model followed by lateral epithelial spread as an explanation of multiple head and neck squamous cell carcinomas. PMID- 12850379 TI - Loss of heterozygosity of a locus in the chromosomal region 17p13.3 is associated with increased cell proliferation in astrocytic tumors. AB - We had previously reported that loss of heterozygosity (LOH) of the D17S379 locus on 17p13.3 was significantly more frequent in high-grade gliomas (anaplastic astrocytoma, AA; glioblastoma multiforme, GBM) than in those of a low-grade diffuse astrocytoma (DA); however, this was independent of alterations at the TP53 locus, We also showed that LOH of D17S379 was associated with positive staining for p53 protein on immunohistochemistry, but LOH of the TP53 gene had no such association. In this work we show that cell proliferation as determined by MIB-1 labeling index (LI) was significantly higher in tumors with LOH of D17S379 than those with no LOH (NLOH). In accord with our previous results, p53 protein immunopositivity was also associated with increased MIB-1 LI; however, we observed no such association of LI with TP53 LOH. The results further confirm that alteration of one or more putative tumor suppressor loci at 17p13.3 is associated with increased proliferation in astrocytic tumors. PMID- 12850380 TI - Intratumoral genomic heterogeneity in advanced head and neck cancer detected by comparative genomic hybridization. AB - Little is known about the extent of intratumoral genetic heterogeneity in head and neck squamous cell carcinoma (HNSCC). We therefore examined 79 stage III and IV primary HNSCCs (P) and matched lymph node metastases (M) for over- and underrepresentation of specific chromosome regions by comparative genomic hybridization (CGH). The overall ratio of gains and losses was higher in metastases than in primary tumors (4/1 vs. 2.5/1). Gains of 3q (78.1% P vs. 87.5% M) and 11q (78.1% P vs. 62.5% M) and deletions of 3p (43.8% P vs. 34.4% M) and 9p (31.3% P vs. 15.6% M) were most frequently detected. The highest rate of intratumoral discordance was observed for primary tumors and corresponding metastases (32.8%) compared with matched pairs of two metastases (26.5%) and of two anatomically distinct sides of one primary tumor (24.3%). Furthermore, the discordance rate was dependent on the primary tumor site (oral cavity 49.2%, oropharynx 31%, hypopharynx 30.3%, and larynx 27.3%). In some tumors, the extent of genomic discordance argues against a monoclonal origin. In conclusion, we found a high individual variation of intratumoral genomic heterogeneity depending on the localization and selection of matched pairs. These findings are of specific importance in view of establishing prognostic markers. PMID- 12850381 TI - A novel translocation der(4)t(1;4)(q21;q35) and a marker chromosome in a case of myelodysplastic syndrome. PMID- 12850382 TI - A case of acute myelogenous leukemia: myelodysplastic syndrome with t(2;11)(p21;q23) without MLL rearrangement. PMID- 12850383 TI - Measurement of plasma endothelin-1 in experimental hypertension and in healthy subjects. AB - BACKGROUND: Endothelin-1 is an endothelium-derived potent vasoconstrictor peptide of 21 amino acids. To establish reference values in different models of hypertension and in human subjects an assay for plasma immunoreactive endothelin 1 (ET-1) was optimized. METHODS: ET-1 is extracted by acetone from 1 mL of plasma and subjected to a sensitive enzyme-linked immunosorbent assay. RESULTS: The detection limit for plasma ET-1 is 0.05 fmol/mL. Mean recoveries of the 1, 2, 5, and 10 fmol of ET-1 added to 1 mL of plasma were 66%, 75%, 85%, and 92%, respectively. Within- and between-assay coefficients of variation were < or =12% and < or =10%, respectively. Assay accuracy was demonstrated by consistent recoveries of added ET-1 over the entire physiologic range of plasma concentrations and by the linearity of ET-1 concentrations measured in serially diluted plasma extracts (r = 0.99). No ET-1 was detected when albumin buffer was extracted instead of plasma. Using this method, we found increased ET-1 levels in plasma of three experimental rat models of hypertension: stroke prone spontaneously hypertensive rats (SP-SHR), deoxycorticosterone acetate-salt hypertensive rats, and one kidney-one clip hypertensive rats. In contrast, plasma ET-1 levels of SHR were half those of normotensive Wistar rats. In two kidney-one clip hypertensive rats, plasma ET-1 concentrations were not different from those found in sham-operated control rats. Plasma ET-1 concentrations of 37 healthy men were 0.85 +/- 0.26 fmol/ml (mean +/- SD). CONCLUSIONS: The present assay reliably measures ET-1 levels in rat and human plasma. It allows to discriminate between different forms of hypertension with high or low circulating levels of ET-1. PMID- 12850384 TI - Urinary tetrahydroaldosterone as a screening method for primary aldosteronism: a comparative study. AB - BACKGROUND: The major aldosterone metabolite 3 alpha,5 beta tetrahydroaldosterone reflects up to 45% of the aldosterone secretion. Its 24-h urinary excretion is likely to provide an accurate index of the daily aldosterone production and to be an indicator for primary aldosteronism (PA). METHODS: In a prospective study, the validity of tetrahydroaldosterone as a screening test for PA was evaluated in comparison to serum potassium, plasma aldosterone, plasma renin activity, plasma aldosterone/renin activity ratio (PARR), as well as 24-h urinary aldosterone-18 glucuronide and free aldosterone. A total of 111 normotensive individuals, 412 PA patients and 1453 essential hypertensive patients, were studied. The effect of blood sampling technique on potassium level was also investigated. RESULTS: Tetrahydroaldosterone differentiated PA from essential hypertension with a sensitivity of 96% and a specificity of 95%. The sensitivity was 89% for plasma aldosterone, 87% for free aldosterone, 85% for PARR, 71% for aldosterone-18 glucuronide and 51% for renin activity. Specificities varied between 91% and 85%. The combined use of the parameters plasma aldosterone > or =9.0 ng/dL and PARR > or =25 resulted in a sensitivity of 82% and specificity of 95%. Forearm exercise proved to be a source of erroneous elevations of potassium sufficient to obscure the suspicion of PA. CONCLUSION: The data suggest that tetrahydroaldosterone is the most reliable screening test for PA. Tetrahydroaldosterone determination in combination with aldosterone-18-glucuronide and free aldosterone increases diagnostic specificity for PA. Potassium, renin, plasma aldosterone, and basal PARR are inadequate screening procedures because they are subject to high rates of false-positive and false-negative results. PMID- 12850385 TI - Reduced vagal activity in salt-sensitive subjects during mental challenge. AB - BACKGROUND: Salt-sensitive normotensive men exhibit an enhanced pressor and heart rate (HR) response to mental stress. Stress-induced HR acceleration may result from sympathetic activation or vagal withdrawal. We studied the importance of vagal withdrawal for the increased stress responsiveness of salt-sensitive subjects. METHODS: We studied cardiovascular reactivity to mental challenge in 17 salt-sensitive healthy white male students and 56 salt-resistant control subjects who were comparable with respect to age, body mass index, and physical fitness. Salt sensitivity was determined by a 2-week dietary protocol (20 mmol v 240 mmol sodium/day). Mental stress was induced by a computerized information-processing task (manometer test). Electrocardiogram and finger blood pressure (BP; Finapres, Ohmeda, Louisville, CO) were registered continuously to determine HR and interbeat-interval length. Time and frequency domain (spectral power) based measures of respiratory-related heart rate variability (HRV) were calculated to estimate vagal cardiac control; diastolic BP reactivity was assessed to estimate peripheral sympathetic effects. RESULTS: Stress-induced increase in HR was higher in salt-sensitive than in salt-resistant subjects. Salt-sensitive subjects, in comparison to salt-resistant subjects, showed significantly reduced respiratory related HRV during baseline and mental stress conditions (P <.01). The increase in diastolic BP during mental challenge was significantly greater in salt sensitive subjects (P <.05). CONCLUSIONS: Our findings suggest reduced vagal and increased sympathetic tone during mental challenge in salt-sensitive subjects. Altered autonomic nervous system function may contribute to later development of hypertension in salt-sensitive individuals. PMID- 12850386 TI - Is chronic inflammation a determinant of blood pressure in the elderly? AB - BACKGROUND: Previous studies have shown that a rise in blood pressure (BP) causes chronic inflammation of the endothelium which, in turn, may be responsible for further damage of endothelium and worsening of BP control. On the other hand, several metabolic abnormalities such as dyslipidemia, hyperinsulinemia/insulin resistance, diabetes, and obesity causes inflammation followed by a later rise in arterial BP. We investigated the role of chronic inflammation in the modulation of BP independently of other traditional cardiovascular risk factors and atherosclerotic lesions. METHODS: A total of 537 aged subjects, selected from the whole population of the INCHIANTI cohort, were enrolled. All subjects underwent plasma insulin, glucose, interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-1 beta (IL-1 beta), interleukin-1 receptor antagonist (IL-1ra), C reactive protein, and tumor necrosis factor-alpha (TNF-alpha) levels determination. The IL-6-174 C/G promoter polymorphism was also evaluated. RESULTS: After adjusting for age, sex, insulin resistance syndrome score, and severity of carotid atherosclerosis, serum IL-1 beta (P <.001), IL-1ra (P <.005) concentration and the insulin resistance syndrome score (P <.001) were the only predictors of diastolic BP. Indeed, age (P <.001), insulin resistance syndrome score (P =.05), IL-1 beta (P <.05), and severity of carotid atherosclerosis (P <.05) were the only significant predictor of systolic BP. CONCLUSION: These results suggest that chronic inflammation may play a role in the modulation of arterial BP. PMID- 12850387 TI - VALUE trial: Long-term blood pressure trends in 13,449 patients with hypertension and high cardiovascular risk. AB - BACKGROUND: The Valsartan Antihypertensive Long-term Use Evaluation (VALUE) study compares cardiovascular outcomes in 15,314 eligible patients from 31 countries randomized to valsartan or amlodipine-based treatment. METHODS: The blood pressure (BP) trends are analyzed in 13,449 of VALUE study patients who had baseline BP and 24 months BP and treatment data. RESULTS: In a cohort of 12,570 patients, baseline 24 and 30 months BP, but not 30 months treatment data, were available. Of 13,449 patients, 92% (N = 12,398) received antihypertensive therapy at baseline. The baseline BP was 153.5/86.9 mm Hg in treated compared to 168.1.8/95.3 mm Hg in 1051 untreated patients. After 6 months both groups had indistinguishable BP values. At 12 months the BP decreased to 141.2/82.9 mm Hg (P <.0001 for systolic BP and diastolic BP versus baseline), at 24 months to 139.1/80 mm Hg (P <.0001 v 12 months), and to 138/79 mm Hg at 30 months (P <.0001 v 24 months). The systolic BP control (<140 mm Hg) at 30 months increased from 21.9% at baseline to 62.2%, the diastolic BP (< 90 mm Hg) from 54.2% to 90.2% and the combined control (<140 and <90 mm Hg) from 18.9% to 60.5%. At 24 months 85.8% of patients were on protocol drugs: monotherapy = 39.7%, added hydrochlorothiazide = 26.6%, add-on drugs = 15.1%, and protocol drugs in nonstandard doses = 4.3%. CONCLUSIONS: The achieved BP control exceeds values reported in most published large-scale trials. The VALUE study is executed in regular clinical settings and 92% of the patients received antihypertensive drugs at baseline. When an explicit BP goal is set, and a treatment algorithm is provided, the physicians can achieve better control rates than in their regular practice. PMID- 12850388 TI - Adolescent blood pressure and blood pressure tracking into young adulthood are related to subclinical atherosclerosis: the Atherosclerosis Risk in Young Adults (ARYA) study. AB - BACKGROUND: Increased blood pressure (BP) in young adulthood is associated with cardiovascular morbidity and mortality. Longitudinal studies of patients at young ages are, however, limited. Our aim was to study the relationships of adolescent BP and tracking of BP into young adulthood with subclinical atherosclerosis, as assessed by carotid intima-media thickness (CIMT), at the age of 28 years. METHODS: The Atherosclerosis Risk in Young Adults (ARYA) study comprises of a community-based sample of 750 subjects aged 27 to 30 years. In the 352 men and 398 women, at least one BP measurement was recorded at a mean age of 13 years in school health records. Recently, all participants completed a questionnaire on cardiovascular risk factors, had a fasting blood sample drawn, and underwent an ultrasound examination of both common carotid arteries to assess CIMT. RESULTS: Linear regression showed that adolescent systolic BP was associated with thickening of the intima-media (an increase of 7.5 microm in CIMT per standard deviation increase in systolic BP; 95% CI 4.3 to 10.6). Similar relations were found for pulse pressure and mean arterial pressure. When sex, age, and body mass index at adolescence and young adulthood and adult BP were taken into account, the relations attenuated, but for pulse pressure they remained statistically significant. Furthermore, subjects who tracked in the highest systolic BP and pulse pressure levels from adolescence into young adulthood showed the thickest CIMT. CONCLUSION: Our findings strengthen the notion that elevated BP at adolescence and a relative increase in BP from adolescence to adulthood unfavorably affect cardiovascular risk, as indicated by increased CIMT. PMID- 12850389 TI - Prevalence of left ventricular hypertrophy in patients with mild hypertension in primary care: impact of echocardiography on cardiovascular risk stratification. AB - BACKGROUND: Left ventricular hypertrophy (LVH) is an important predictor of cardiovascular risk, and its detection contributes to risk stratification. The aims of the present study were to estimate the prevalence of echocardiographic LVH and to evaluate the influence of echocardiography (ECHO) on cardiovascular risk stratification in hypertensive patients presenting in primary care. METHODS: In this cross-sectional study, 250 patients recently diagnosed with mild hypertension underwent clinical evaluation including electrocardiography (ECG), microalbuminuria measurement, 24-h blood pressure monitoring and ECHO. Level of cardiovascular risk was stratified, initially using routine procedures including ECG to assess target organ damage and then again after detection of LVH by ECHO. RESULTS: The frequency of echocardiographic LVH was 32%, substantially higher than that detected by ECG (9%). Initial cardiovascular risk stratification yielded the following results: 30% low risk, 49% medium risk, 16% high risk, and 5% very high risk subjects. The detection of LVH by ECHO provoked a significant change in the risk strata distribution, particularly in those patients initially classified as being at medium risk. In this group, 40% of subjects were reclassified as high risk subjects according to ECHO information. The new classification was as follows: 23% low risk, 30% medium risk, 42% high risk, and 5% very high risk subjects. CONCLUSIONS: A substantial proportion of mildly hypertensive patients presenting in primary care have LVH determined by ECHO. Our results suggest that this procedure could significantly improve cardiovascular risk stratification in those patients with multiple risk factors, but no evidence of target organ damage by routine investigations. PMID- 12850390 TI - Determinants of exercise capacity in hypertensive patients: new insights from tissue Doppler echocardiography. AB - BACKGROUND: Determinants of exercise capacity have not been clearly defined in hypertensive patients. Doppler tissue imaging (DTI), a recently developed echocardiographic modality, is useful for evaluating left ventricular (LV) function regardless of loading conditions. The present study was performed to identify the determinants of exercise capacity in hypertensive patients using DTI. METHODS: We studied 48 hypertensive patients (26 men; mean age, 57 +/- 9 years) with normal LV ejection fraction (>50%). Conventional echocardiographic indexes were measured before and after symptom-limited treadmill exercise. Also, longitudinal systolic (S(L)') and diastolic myocardial velocities (E(L)' and A(L)') and radial velocities (S(R)', E(R)', A(R)') were measured using DTI with the sample volume at the LV basal posterior wall. RESULTS: Mean exercise time was 525 +/- 135 sec. Mean LV ejection fraction was 61% +/- 6%. Mean LV mass index was 110.9 +/- 24.4 g/m(2). On univariate analysis, S(L)', E(L)', A(L)' and percentage increase of S(R)' velocity after exercise showed significant correlation with exercise time. However, only E(L)' and percentage increase of S(R)' remained significant after multivariate analysis. CONCLUSIONS: Left ventricular contractile reserve and relaxation property are important cardiac factors determining exercise capacity. Early diastolic longitudinal myocardial velocity might be useful for identifying hypertensive patients with poor exercise capacity. PMID- 12850391 TI - Dissociation of coronary artery contractile hyperreactivity from hypertension. AB - BACKGROUND: Both coronary artery contractile hyperreactivity and hypertension are associated with increased coronary artery disease. It is not known how coronary artery contractile hyperreactivity relates to hypertension. The current study tests the hypothesis that coronary artery contractile hyperreactivity can be dissociated from hypertension and therefore may contribute to the etiology of CAD independent of hypertension. METHODS: The contractile responses to 5 hydroxytryptamine (5-HT) and guanosine triphosphate (GTP) were determined in intact (nonpermeabilized) and alpha-toxin-permeabilized coronary artery strips and small mesenteric artery strips isolated from four rat strains: spontaneously hypertensive rats (SHR), Wistar-Kyoto rats (WKY), WKY-derived hypertensive rats (WKHT), and WKY-derived hyperactive rats (WKHA). RESULTS: The SHR and WKHT were hypertensive, whereas the WKY and WKHA subjects were normotensive. The coronary artery contractile reactivity to 5-HT was significantly higher in SHR when compared with WKY. The coronary artery contractile reactivity was of similar magnitude in WKHA and WKHT and was intermediate between that of SHR and WKY rats. GTP-induced Ca(2+) sensitization of contractions were significantly greater in SHR than in WKHT, WKHA, and WKY; in comparison, no significant difference was found among WKHT, WKHA, and WKY. In contrast to the findings in coronary arteries, there was no significant difference in 5-HT-induced contractions in small mesenteric artery strips isolated from SHR and WKY. CONCLUSIONS: Coronary artery contractile reactivity to 5-HT does not correlate entirely with blood pressure (BP) values. In addition, G-protein-mediated Ca(2+) sensitization of contraction was increased and contributed to the coronary artery contractile hyperreactivity in SHR. PMID- 12850392 TI - N-acetylcysteine improves nitric oxide and alpha-adrenergic pathways in mesenteric beds of spontaneously hypertensive rats. AB - BACKGROUND: The aim of this study was to assess the effects of N-acetylcysteine (NAC) on nitric oxide and adrenergic pathways in mesenteric artery from spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). METHODS: Rats were treated with 4 g x kg(-1) x day(-1) of NAC during 4 weeks or mesenteric beds were treated with 10 mmol/L of NAC during 20 min. RESULTS: In conscious rats, the NAC treatment produced a significant reduction of mean arterial pressure (MAP) and heart rate in SHR (P <.001). N(omega)-nitro-L-arginine methyl ester (L-NAME) caused a MAP increase in NAC-treated SHR of magnitude similar to that in WKY, which was significantly higher than that observed in control untreated SHR (P <.05). Chronic treatments with NAC improved the maximal relaxation of mesenteric arteries to A23187 in SHR (P <.001). Acute NAC treatment in vitro induced a vasodilation in Phe preconstricted arteries (P <.001) that was stronger in SHR than in WKY (P <.05) and was not abolished by L-NAME. The vasoconstrictory response and increases in inositol phosphate production induced by superoxide anion were attenuated by NAC treatment through its superoxide scavenging properties. In contrast, chronic and acute NAC treatments did not alter the vasodilatory response to beta-adrenergic receptor stimulation. CONCLUSIONS: The increase in NO-mediated vasodilator tone and the possible decrease in adrenergic vasoconstriction induced by NAC treatment in SHR could explain the hypotensive effect of NAC in this model of hypertension. PMID- 12850393 TI - Comparative study of sinoaortic denervated rats and spontaneously hypertensive rats. AB - BACKGROUND: Both hypertension and high blood pressure variability (BPV) are involved in cardiovascular damage. This comparative study was designed to explore the possible effects of both of these phenomena on the cardiovascular system. METHODS: The high BPV model of 16-week sinoaortic denervated (SAD) rats and the hypertension model of spontaneously hypertensive rats (SHR) were used for comparison at the same age of 26 weeks. The comparison was focus on hemodynamics, cardiovascular hypertrophy, and hemodynamic responses to ketanserin. Linear regression analysis was performed to study the role of hemodynamics in cardiovascular hypertrophy. RESULTS: In SHR, hypertension was accompanied by a moderately high BPV, whereas in SAD rats, substantially high BPV existed alone, without hypertension. Left ventricular hypertrophy was severe in SHR but was mild in SAD rats. Aortic hypertrophy was present in SAD rats but was absent in SHR. In SAD rats, the hypertrophy was correlated with BPV but not with blood pressure (BP) level. However, in SHR, hypertrophy was correlated with both BP and BPV level. The BP-lowering effect of ketanserin was comparable in both models, whereas its BPV-lowering effect was greater in SAD rats than in SHR. This hypersensitivity was associated with basal BPV level in SAD rats. CONCLUSIONS: These results indicate that hypertension may be more important than high BPV in causing left ventricular hypertrophy, and that the aorta may be more sensitive to substantially high BPV. PMID- 12850394 TI - Secondary hypertension due to a renin-producing teratoma. AB - Twenty cases of extrarenal renin-secreting tumors have been reported, but this is the first case of a renin-producing teratoma. The patient was a 17-year-old African American girl who presented with hypertension and hypokalemia, and who was documented to have a plasma aldosterone-to-renin activity ratio consistent with secondary aldosteronism. Computed tomography demonstrated a pelvic tumor suspicious for a teratoma. With no other apparent etiology for the secondary aldosteronism, the teratoma was suspected to be an extrarenal renin-secreting tumor. This was confirmed after surgery by pathologic evaluation, and significantly reduced requirements for antihypertensive medication and potassium supplementation. PMID- 12850395 TI - Differential effects of lercanidipine and nifedipine GITS on plasma norepinephrine in chronic treatment of hypertension. AB - This study aimed to compare the effects of two long-acting dihydropyridine calcium channel blockers (CCBs) with different pharmacologic properties, lercanidipine and nifedipine Gastro-Intestinal Therapeutic System (GITS), in the chronic treatment of essential hypertension. After a 4-week placebo run-in period, 60 patients of both sexes were randomly treated with lercanidipine 10 to 20 mg or nifedipine GITS 30 to 60 mg taken orally for 48 weeks, according to a double-blind, parallel group design. For the first 4 weeks of treatment, the lowest dose of each drug was used, followed by higher doses if diastolic blood pressure (BP) was >90 mm Hg. At the end of the placebo period and after 4, 8, 12, 24, and 48 weeks of active treatment BP, heart rate (HR), and plasma norepinephrine (NE) levels were assessed. Lercanidipine and nifedipine GITS similarly reduced BP values after 48 weeks (-21.7/15.9 mm Hg and -20.7/14.6 mm Hg, respectively, both P <.001 v placebo), with no change in HR. Despite the similar lack of effect on HR, the two drugs displayed different influences on plasma NE, which was significantly increased by nifedipine GITS (+56 pg/mL, P <.05 v placebo) but not by lercanidipine. These findings suggest that 1) sympathetic activation occurs during chronic therapy with nifedipine GITS but not with lercanidipine, which might be related to the different pharmacologic characteristics of the two CCBs at the doses evaluated; and 2) nifedipine GITS seems to activate peripheral but not cardiac sympathetic nerves, consistent with differing regulation of cardiac and peripheral sympathetic activity. PMID- 12850396 TI - Control of arterial pressure by angiotensin II and nitric oxide at the onset of diabetes. AB - We have reported that the induction of diabetes in N(omega)-nitro-L-orginine methyl ester (L-NAME)-infused rats causes significant hypertension that is associated with increased plasma renin activity. This study tested the role of angiotensin II (Ang II) by clamping it chronically at baseline levels. The clamp consisted of an intravenous infusion of enalapril (10 mg/kg/d), which decreased mean arterial pressure (MAP) by approximately 20 mm Hg after 3 days, and adding chronic Ang II at 4 ng/kg/min, which restored MAP to normal. Chronic L-NAME infusion increased MAP to 127 +/- 1 and 132 +/- 2 mm Hg in normal and clamped rats, respectively, and induction of diabetes (streptozotocin) increased MAP progressively in normal rats to 161 +/- 8 mm Hg by day 12, whereas MAP in the clamped rats decreased progressively to 98 +/- 5 mm Hg by day 12. In non-L-NAME rats, MAP averaged 95 +/- 1 and 91 +/- 1 mm Hg for normal and clamped groups, respectively, before diabetes, and MAP was 10 to 13 mm Hg lower in the clamped versus normal rats midway through the diabetic period. This suggests that Ang II is important for maintaining blood pressure at the onset of diabetes, possibly to compensate for renal volume losses. Angiotensin II also is required for the hypertension caused by induction of diabetes in rats with chronic blockade of nitric oxide synthesis, but whether this is due to increased volume sensitivity in L-NAME-treated, vasoconstricted rats remains to be determined. PMID- 12850397 TI - Association between transforming growth factor-beta and hypertension. AB - Discordant findings are reported on the left ventricular transforming growth factor-beta(1) (TGF-beta(1)) mRNA levels in various rat models. Left ventricular TGF-beta(1) mRNA levels did not differ between spontaneously hypertensive rats (SHR) and normal rats, between deoxycorticosterone (DOCA)-salt and sham-operated hypertensive rats, but were increased in stroke-prone spontaneously hypertensive rats (SHRSP) and in post-myocardial infarction (MI) rats. Renal cortical TGF beta(1) mRNA levels were, however, higher in DOCA-salt hypertensive rats. Angiotensin II subtype 1 receptor antagonism (AT(1)R) and angiotensin converting enzyme inhibition (ACEI) decreased left ventricular and vascular smooth muscle TGF-beta(1) mRNA levels in SHR and renal TGF-beta(1) mRNA in DOCA-salt hypertensive rats and in SHRSP. In post-MI rats ventricular TGF-beta(1) mRNA decreased by AT(1)R antagonism. In essential hypertensive patients, TGF-beta(1) protein as well as TGF-beta(1) mRNA levels are hyperexpressed. The TGF-beta(1) overproduction in hypertension can be attributed to various factors such as elevated angiotensin II, increased systemic blood pressure (BP) per se, increased fluid shear stress and a differential expression of TGF-beta(1) linked to DNA polymorphism in the promoter. The Arg(25) polymorphism in the TGF-beta(1) gene is associated with higher BP. A higher plasma TGF-beta(1) concentration is found in hypertensive patients with microalbuminuria and left ventricle hypertrophy. In these patients, AT(1)R antagonism and ACEI reduced these plasma TGF-beta(1) levels significantly. PMID- 12850398 TI - Abnormalities in blood pressure regulation in a patient with Parkinson's disease. PMID- 12850399 TI - Sodium and blood pressure: no data dredging, please! PMID- 12850400 TI - Guidelines for international forensic bio-archaeology monitors of mass grave exhumations. AB - The systematic exhumation of mass graves is becoming a frequent occurrence globally. Historically, it has been mostly anthropologists with their particular expertise in osteology that have been engaged by non-governmental organizations (NGO) to monitor mass grave exhumations and postmortem examinations, conducted by a host country or international forensic team, to ensure competence and a concern for justice. The excavation of graves and examination of their contents for the purposes of personal identification of victims and/or collection of evidence for prosecution require the creation of standards that meet international concerns for forensic investigation of the highest quality. However, many anthropologists are not experienced with large sites; do not have much, if any, expertise in archaeology; and are not equipped by training to assess the quality of a forensic pathologist's autopsy. This contribution is directed to both the NGO and the bio archaeologist who are involved in the exhumation of mass graves. The experience and skills appropriate for the bio-archaeological monitor are outlined; similarly, the policies and standard operating procedures (SOP's) of the NGO that will enable the bio-archaeologist to perform their task are detailed. It is becoming increasingly clear that how a grave site was created, filled, and concealed, along with subsequent processes of site formation (e.g. slumping, robbing, animal scavenging and taphonomic alteration), require the expertise of a forensic archaeologist to discover and record site complexity along with associated forensic evidence. Similarly, the bodies themselves which are often skeletonized, fragmented and commingled pose challenges for the anthropologists and pathologists whose postmortem examinations must be critically evaluated. A model protocol for observations to make at the site as well as at the mortuary facility is provided to guide the monitor and to provide a structure for reports which are of standardized content suitable for international agencies concerned with mass grave investigation. PMID- 12850401 TI - Morphometric investigation of emphysema aquosum in the elderly. AB - We compared the lungs of six drowned and six non-drowned persons over 70 to determine whether there is evidence of acute overinflation in drowning that can be distinguished from senile lung emphysema. All left lungs underwent intrathoracic formalin fixation to preserve their state of insufflation. To assess the effects of lung collapse, all right lungs were sectioned as usual and then immersed in formalin. After histological processing, microphotography and image processing of 12 specimens per corpse, 50 binary images of each specimen were measured by computerized morphometry. Intrathoracic fixation resulted in significantly less tissue area (and more airspace) in the left than in the right lungs of both groups. Comparing both groups' left lungs revealed that the interalveolar septa were thinner and the area occupied by connecting nodes smaller in drowning; these single nodes also tended to be smaller. There was a tendency for less alveolar tissue area per image in drowning than in control lungs and for narrow tissue structures to comprise a higher percentage of both the total tissue area and total tissue perimeter per image. We conclude that there is morphological evidence of acute overinflation even in senile lungs, but this is masked by postmortem lung collapse as suggested by the overlapping values of the right lungs. Support to the diagnosis of drowning among drowned elderly patients can only be available in lungs subjected to intrathoracic postmortem fixation. PMID- 12850402 TI - A molecular genetic approach for forensic animal species identification. AB - This study investigated potential markers within chromosomal, mitochondrial DNA (mtDNA) and ribosomal RNA (rRNA) with the aim of developing a DNA based method to allow differentiation between animal species. Such discrimination tests may have important applications in the forensic science, agriculture, quarantine and customs fields. DNA samples from five different animal individuals within the same species for 10 species of animal (including human) were analysed. DNA extraction and quantitation followed by PCR amplification and GeneScan visualisation formed the basis of the experimental analysis. Five gene markers from three different types of genes were investigated. These included genomic markers for the beta-actin and TP53 tumor suppressor gene. Mitochondrial DNA markers, designed by Bataille et al. [Forensic Sci. Int. 99 (1999) 165], examined the Cytochrome b gene and Hypervariable Displacement Loop (D-Loop) region. Finally, a ribosomal RNA marker for the 28S rRNA gene optimised by Naito et al. [J. Forensic Sci. 37 (1992) 396] was used as a possible marker for speciation. Results showed a difference of only several base pairs between all species for the beta-actin and 28S markers, with the exception of Sus scrofa (pig) beta-actin fragment length, which produced a significantly smaller fragment. Multiplexing of Cytochrome b and D-Loop markers gave limited species information, although positive discrimination of human DNA was evident. The most specific and discriminatory results were shown using the TP53 gene since this marker produced greatest fragment size differences between animal species studied. Sample differentiation for all species was possible following TP53 amplification, suggesting that this gene could be used as a potential animal species identifier. PMID- 12850403 TI - Into the decomposed body-forensic digital autopsy using multislice-computed tomography. AB - It is impossible to obtain a representative anatomical documentation of an entire body using classical X-ray methods, they subsume three-dimensional bodies into a two-dimensional level. We used the novel multislice-computed tomography (MSCT) technique in order to evaluate a case of homicide with putrefaction of the corpse before performing a classical forensic autopsy. This non-invasive method showed gaseous distension of the decomposing organs and tissues in detail as well as a complex fracture of the calvarium. MSCT also proved useful in screening for foreign matter in decomposing bodies, and full-body scanning took only a few minutes. In conclusion, we believe postmortem MSCT imaging is an excellent vizualisation tool with great potential for forensic documentation and evaluation of decomposed bodies. PMID- 12850404 TI - Tissue distribution of trichloroethylene in a case of accidental acute intoxication by inhalation. AB - This article describes the toxicological findings in a fatality due to an accidental inhalation of trichloroethylene which took place during wall coating of a poorly ventilated well using trichloroethylene. The man was wearing protective clothing and a mouthmask with adsorbent. He was found dead on the floor of the well 5h after descending. Trichloroethylene was added to the mortar to enhance drying. Identification and quantitation of trichloroethylene in the postmortem samples (blood, lung, liver, kidney, stomach content and bile) and identification of its metabolite trichloroacetic acid in urine was performed using static headspace gas chromatography with mass spectrometric detector. The compounds were separated on a CP-SIL 5CB Low Bleed/MS column using n-butanol as internal standard. The method was linear over the specific range investigated, and showed an accuracy of 104% and an intra-day precision of 11%. Trichloroethylene concentrations of 84mg/l in subclavian blood, 40mg/l in femoral blood, 72mg/kg in liver, 12mg/kg in kidney, 78mg/kg in stomach content, 104mg/l in bile and 21mg/kg in lung were found. Trichloroacetic acid was identified in the urine. PMID- 12850405 TI - Mixed drug intoxication involving zaleplon ("Sonata"). AB - Zaleplon ("Sonata") is a pyrazolopyrimidine derivative approved for use in the United States for the treatment of insomnia. To date, there has been little data in the toxicological literature where zaleplon has been implicated as causing a fatal intoxication, either alone or in combination with other drugs. This report documents a case where zaleplon was identified in a suicide by multiple drug ingestion. The following zaleplon concentrations were found: heart blood 2.2mg/l; bile 8.6mg/l and urine 1.4mg/l. Zaleplon was also detected but not quantitated in the kidney and liver. PMID- 12850406 TI - Camera identity from minute film-edge mark characteristics. AB - A case is presented in which a camera recovered from a site of bomb blast was linked to some incriminating film negatives by the characteristic markings existing along a small portion of the edge of the film negatives. PMID- 12850407 TI - Evaluating the informative power of Y-STRs: a comparative study using European and new African haplotype data. AB - Male individuals from Maputo (Mozambique) were sampled and 18 Y-STRs were typed: the nine currently used to define the "minimal haplotype" employed in the European, American and Asian "Y-STR Haplotype Reference Databases", as well as the recently described DYS434, DYS437, DYS438, DYS439, DYS460, DYS461, GATA A10, GATA C4 and GATA H4. Allele and haplotype frequencies were estimated in a sample of 112 individuals, where it was possible to define 101 haplotypes, with an observed haplotype diversity (HD) of 0.9973. Allele diversity varied between 0.0179 and 0.9220, DYS385 showing the highest level of polymorphism and DYS392 the lowest. When considering only the most recent Y-STRs, the degree of diversity varied between 0.4011 (DYS438) and 0.6910 (GATA C4), except for DYS434 and DYS437 where a very low diversity was observed (0.0700 and 0.0526, respectively). When analysing the same 112 individuals for the nine Y-STRs included in the minimal haplotype, 78 haplotypes were distinguished with a corresponding observed diversity of 0.9884, a considerably lower value than those for Northern Portugal (n=208; HD: 0.9925) and Macao (n=63; HD: 0.9990). Concerning all 18 Y-STRs studied in this population, the observed diversity demonstrates their usefulness in forensic applications, with the exception of DYS434, DYS437 and DYS392. However, since the informative power of a marker has to be judged in haplotype context, a simple software, allowing the evaluation of the increase of HD through the addition of any combination of new markers to the minimum haplotype was designed. The statistical approach devised, demonstrates that an increment on HD is more rapidly obtained for the Mozambican database when adding GATA A10 or DYS439, DYS460, GATA C4, DYS461 or GATA H4, in this order, to the minimal haplotype. DYS434, DYS437 and DYS438, in conjunction with all the other 15 Y STRs, do not contribute to an increment on HD. When applying the same approach to an European sample (Northern Portugal), the first three Y-STR choices coincide, but the next order of markers are GATA H4, then DYS437 and finally DYS461. In this sample, DYS434, DYS438 and GATA C4 do not increment HD any further. PMID- 12850408 TI - Skin changes following defibrillation. The effect of high voltage direct current. AB - Previously, electrical injuries have been suggested caused only by the concomitant heat developed during the passage of an electrical current. Recent experimental studies on fully anesthetized pigs and the study of one human case have, however, shown typical electrical alterations. The purpose of the present study was further to evaluate the histology of electrically induced changes in the skin in humans. In addition, supplementary in vivo methods for evaluation of skin changes as high-frequency ultrasound and Raman spectroscopy were used. The skin of 11 patients treated with a defibrillation of the heart was examined for macroscopic changes, the skin of eight of them also for histologic changes and for changes observable via supplementary methods. Immediately and 7 days after the defibrillation, fractions of a narrow red ring were observed along the periphery of the tin-foil electrode. Epidermis showed signs previously observed following electrical influence: segmental alterations often related to the openings of sweat ducts, darkstaining or "empty" nuclei and homogeneous cytoplasm, eosinophilic or pale. Dermis did not show the specific sign of electrical influence: deposits of calcium salts on dermal fibres, neither via histologic examination nor via high-frequency ultrasonography and Raman spectroscopy. Fractions of a narrow red ring along the periphery of the electrode showing histological signs of electric influence in epidermis thus appear to be characteristic of high voltage electrical injury. PMID- 12850409 TI - Criteria for homicide and suicide on victims of extended suicide due to sharp force injury. AB - This report is about the findings in association with the extended suicides of nine victims killed by sharp force. All victims were killed by sharp force. The perpetrators were predominantly the parents, the victims their children. Regarding the criteria for differentiating self-inflicted injuries from injuries inflicted by another person, the victims' injuries presented patterns usually found solely in suicides. Thus eight of nine cases presented tentative and hesitation injuries, in three of five cases areas of injury covered by clothing had been exposed beforehand. Despite extremely narrow intercostal spaces in children, injuries to the bones in thoracic stabbing were avoided more often than not (four of seven cases). Only the criterion "defence injury" occurred nearly as often as in homicide victims (three of nine cases). The psychopathology of extended suicide can explain this pattern. The perpetrator's motive is characterised by his pseudoaltruistic belief to save the loved ones from a world that is in his opinion unacceptable. A fusion or integration of the victim into the perpetrator's own self is based on an identity problem. Physical interrelation of forces between perpetrator and victim restricts the victim in his defence and presents an important prerequisite for acquiring the patterns of described injuries. PMID- 12850410 TI - Right ventricular damage due to pulmonary embolism: examination of the number of infiltrating macrophages. AB - To investigate the pathological changes in the heart induced by pulmonary embolism, 20 autopsy cases of pulmonary embolism and 10 control cases of acute death from traumatic injury were examined. Adding to the routine hematoxylin eosin (HE) staining, immunostaining with CD68 pan-macrophage marker was performed on the specimens obtained from both right and left ventricular walls. The number of macrophages was counted semi-quantitatively in 100 random high-power fields (HPF). Although typical pathological findings of myocardial infarction was not observed in any of the cases, 16 of the 20 pulmonary embolism cases showed an increase in the number of macrophages, mainly in the right ventricular wall. Four cases showed massive macrophage infiltration in the entire right ventricular wall. It is speculated that ischemia due to pulmonary embolism may be connected to its pathogenesis. PMID- 12850411 TI - The incidence of drugs in drivers killed in Australian road traffic crashes. AB - The incidence of alcohol and drugs in fatally injured drivers were determined in three Australian states; Victoria (VIC), New South Wales (NSW) and Western Australia (WA) for the period of 1990-1999. A total of 3398 driver fatalities were investigated which included 2609 car drivers, 650 motorcyclists and 139 truck drivers. Alcohol at or over 0.05 g/100ml (%) was present in 29.1% of all drivers. The highest prevalence was in car drivers (30.3%) and the lowest in truckers (8.6%). WA had the highest rate of alcohol presence of the three states (35.8%). Almost 10% of the cases involved both alcohol and drugs. Drugs (other than alcohol) were present in 26.7% of cases and psychotropic drugs in 23.5%. These drugs comprised cannabis (13.5%), opioids (4.9%), stimulants (4.1%), benzodiazepines (4.1%) and other psychotropic drugs (2.7%). 8.5% of all drivers tested positive for Delta(9)-tetrahydrocannabinol (THC) and the balance of cannabis positive drivers were positive to only the 11-nor-Delta(9) tetrahydrocannabinol-9-carboxylic acid (carboxy-THC) metabolite. The range of THC blood concentrations in drivers was 0.1-228 ng/ml, with a median of 9 ng/ml. Opioids consisted mainly of morphine (n=84), codeine (n=89) and methadone (n=33), while stimulants consisted mainly of methamphetamine (n=51), MDMA (n=6), cocaine (n=5), and the ephedrines (n=61). The prevalence of drugs increased over the decade, particularly cannabis and opioids, while alcohol decreased. Cannabis had a larger prevalence in motorcyclists (22.2%), whereas stimulants had a much larger presence in truckers (23%). PMID- 12850412 TI - Smothering of children older than 1 year of age-diagnostic significance of morphological findings. AB - A homicide by smothering may leave unobtrusive or even no injuries if the victim is not able to struggle. This is especially true for infants up to 12 months of age. The differentiation between the sudden infant death syndrome and smothering, by an autopsy alone may be impossible. To establish whether this is different in older children because of their rising capacity for defence six cases of smothering in children >1 year of age were re-examined. The age of the children ranged between 1.5 and 7 years (surviving child). The smothering was inflicted with hands in three cases (including the surviving child), with a pillow in two cases, and the way of smothering remained unknown in one case (no confession of the perpetrator). Depending on the tool used for smothering, abrasions on the facial skin (hands, lesser in cases of smothering by a pillow) and petechiae (pillow/hands) could be seen by external examination. The older the children were, the more injuries could be found. Together with circumstantial evidence all cases could have been established by a thorough autopsy. This is important to prevent further offences against (surviving) siblings. PMID- 12850413 TI - Fatal subarachnoid hemorrhage complicating actinomycotic meningitis. AB - Actinomycosis is caused by Gram-positive Actinomyces species that are part of the normal oral flora with low virulence. We describe a rare case of sudden death of a 48-year-old man with actinomycotic basilar meningitis that was complicated by fatal subarachnoid hemorrhage. Autopsy revealed meningitis at the basilar region of the brain, and histological examination revealed characteristic bacterial aggregates with extensive leukocyte infiltration and severe vasculitis of arteries of this region. Rupture of an artery by severe arteritis was thought to be the cause of the subarachnoid hemorrhage. The probable primary source of infection was found in the left lung. To the best of our knowledge, the complication of subarachnoid hemorrhage has not been reported previously in actinomycotic meningitis. PMID- 12850414 TI - Grouping of Y-STR haplotypes discloses European geographic clines. AB - Y-STR haplotypes are widely studied in Europe and an extensive databasing effort has been conducted (http://www.ystr.org). The distribution of these haplotypes has been considered to present no evidence for substructure at central and southern European level. This picture contrasts with the one that results from Y haplogroups defined by binary markers. This paradox has been solved by admitting that the high STR mutation rate and corresponding recurrence has erased geographic structuration. This explanation prompted us to reanalyse Y-STR haplotypes distribution bearing in mind the commonly admitted model for the generation of diversity in these markers, namely the stepwise mutation model (SMM) and, thus, taking the molecular distance between haplotypes into consideration. Accordingly, we have studied the European distribution of the two most frequent haplotypes in the Iberian Peninsula and their one step neighbours using the European samples deposited in the Y STR database (http://www.ystr.org). For the first group we found a clear-cut decreasing W-E gradient, while for the second the highest frequencies were found in the Iberian Peninsula (3.98% in Portugal and 3.85% in Spain), dropping to 2.88% in France and showing a less well defined SW-NW gradient. Furthermore, we have tested the agreement between haplotype groups and binary markers haplogroups in a random sample of 292 individuals from Northern Portugal. Our results demonstrate that (a) Y-STR haplotype data can be used for wide-scale anthropological approaches disclosing information that has been considered only available through binary markers and (b) forensic use of continental databases needs careful refinement, due to the macro-geographic pattern now evidenced. PMID- 12850415 TI - DNA mixtures in forensic casework: a 4-year retrospective study. AB - Occasionally interpretation guidelines from validation studies are difficult to apply to real forensic casework, especially in the case of mixed samples. Exogenous contamination, an unknown number of contributors or unbalanced proportion of each one in the sample and a varied degree of degradation of the biological materials, contribute to the difficulties in the interpretation of sample profiles. In this paper we have reviewed all the mixed genetic STR profiles encountered in our laboratory over 4 years (1997-2000) and evaluated the problems in the interpretation of the results. From 1547 criminal cases with 2424 samples typed, 163 showed a mixed profile (6.7%). We have observed that occasionally, a mixture appeared in the same sample with one multiplex amplification kit (e.g. Blue) and not with another (e.g. Green). From our results, it can be suggested that technical characteristics of the different fluorochrome groups in the multiplexes override the molecular characteristics of each STR in their capacity to detect mixtures. PMID- 12850416 TI - Analysis of ecstasy tablets: comparison of reflectance and transmittance near infrared spectroscopy. AB - Calibration models for the quantitation of commonly used ecstasy substances have been developed using near infrared spectroscopy (NIR) in diffuse reflectance and in transmission mode by applying seized ecstasy tablets for model building and validation. The samples contained amphetamine, N-methyl-3,4-methylenedioxy amphetamine (MDMA) and N-ethyl-3,4-methylenedioxy-amphetamine (MDE) in different concentrations. All tablets were analyzed using high performance liquid chromatography (HPLC) with diode array detection as reference method. We evaluated the performance of each NIR measurement method with regard to its ability to predict the content of each tablet with a low root mean square error of prediction (RMSEP). Best calibration models could be generated by using NIR measurement in transmittance mode with wavelength selection and 1/x transformation of the raw data. The models build in reflectance mode showed higher RMSEPs using as data pretreatment, wavelength selection, 1/x transformation and a second order Savitzky-Golay derivative with five point smoothing was applied to obtain the best models. To estimate the influence of inhomogeneities in the illegal tablets, a calibration of the destroyed, i.e. triturated samples was build and compared to the corresponding data of the whole tablets. The calibrations using these homogenized tablets showed lower RMSEPs. We can conclude that NIR analysis of ecstasy tablets in transmission mode is more suitable than measurement in diffuse reflectance to obtain quantification models for their active ingredients with regard to low errors of prediction. Inhomogeneities in the samples are equalized when measuring the tablets as powdered samples. PMID- 12850417 TI - FINEX: a Probabilistic Expert System for forensic identification. AB - A series of recent papers have shown how to formulate complex problems of forensic DNA identification inference, such as occur in disputed paternity or criminal identification cases, in terms of Probabilistic Expert Systems (PESs). However, at the present time, general purpose PES software is not particularly well suited to the repetitive tasks of: specifying an appropriate set of marker networks for a specific problem; for editing the many local conditional probability tables; and combining evidence from several genetic markers to evaluate likelihoods. Here, I describe a user-friendly prototype software tool called FINEX developed both to automate such tasks and also to evaluate likelihoods of interest. Ease of use is achieved by a graphical specification language that enables a user to quickly specify a range of forensic DNA problems. I describe the algorithms by which FINEX converts the user input in the graphical specification language and data on observed markers to the Bayesian networks used in PES. PMID- 12850418 TI - Synthesis of standards of the most important markers of Leuckart p methoxymethamphetamine (PMMA). Examination of the influence of experimental conditions and a drug diluent on SPE/TLC profiling. AB - The synthesis of characteristic markers of PMMA obtained by Leuckart method was described. The effectiveness of a procedure of SPE/TLC screening profiling of impurities was studied on the basis of selected impurities. The influence of glucose (a drug diluent) on the profile quality was investigated. The intermediate product (characteristic for the Leuckart synthesis) N-formyl-p methoxymethamphetamine (1) and by-products: N-formyl-p-methoxyamphetamine (2), p methoxyamphetamine (3), N,N-dimethyl-p-methoxyamphetamine (4), (RS) and (RR/SS) diastereoisomers of bis(1-methyl-2-(4-methoxyphenyl)ethyl)amine (meso-5 and rac 5), (RS) and (RR/SS) diastereoisomers of N-methyl-bis(1-methyl-2-(4 methoxyphenyl)ethyl)amine (meso-6 and rac-6), N-methyl-1,3-bis(4 methoxyphenyl)propane-2-amine (7) were synthesized. The substrate p methoxyphenylacetone and the impurities 1 and 4 were used in the study of influence of experimental conditions and glucose on the profiling process and results. The experiments were carried out according to a 2(4) factorial design. The proposed criterions of the profile quality are based on matrix presentation of TLC patterns. They take into account the number of spots revealed, differences between R(f) values and intensity of fluorescence, simultaneously. PMID- 12850419 TI - Profiling of impurities in p-methoxymethamphetamine (PMMA) by means of SPE/TLC method. Examination of the influence of experimental conditions according to 2(4) factorial. AB - In the present paper profiling of impurities in p-methoxymethamphetamine (PMMA) by means of SPE/TLC is reported. PMMA was synthesised by Leuckart procedure. The influence of experimental conditions on the profile quality was investigated. The experiments were carried out according to a 2(4) factorial. The proposed characteristics of the profile quality (optimisation criterions) are based on a matrix presentation of TLC patterns. They take into account, simultaneously, the number of spots revealed, differences between R(f) values and intensity of fluorescence. PMID- 12850420 TI - STR data for PowerPlex 16 System from Neuquen population, SW Argentina. AB - Allele frequencies for the 15 autosomic STR loci included in the PowerPlex 16 System kit (Promega) were estimated from a sample of 111 unrelated individuals living in Neuquen province, southwest of Argentina. Population showed to be in HWE. PMID- 12850421 TI - STR data for PowerPlex 16 System from Buenos Aires population, Argentina. AB - Allele frequencies for the 15 autosomic STR loci included in the PowerPlex 16 System kit (Promega Corp.) were estimated from a sample of 143 unrelated individuals living in Capital Federal and in Buenos Aires Metropolitan Area, Argentina. Population showed to be in HWE. PMID- 12850422 TI - Genetic diversity in four tribal groups of western India: a survey of polymorphism in 15 STR loci and their application in human identification. AB - Genetic diversity at 15 STR loci: 2 pentanucleotide and 13 tetranucleotide STR loci was determined in four highly endogamous tribal groups, viz. Madia-Gond, Mahadeo-Koli, Katkari and Pawara of western India. The distribution of genotypes at studied 15 loci was found in agreement with expected values according to Hardy Weinberg equilibrium. The combined power of discrimination of 15 loci was calculated as 0.80 while combined power of exclusion was observed as 0.53 among the studied four tribal groups. The study demonstrate very low heterozygosity and low power of exclusion of the loci of Powerplex 16 among the selected groups indicating less informativeness of the studied markers in human identification testing. PMID- 12850423 TI - Fatal hepatic failure following accidental tramadol overdose. AB - A case of fatal overdose of tramadol is described, occurring in a 67-year-old man with painful rib fractures who accidentally ingested more than the recommended daily dose. The mode of death was acute liver failure due to fulminant hepatic necrosis. Post-mortem toxicology was negative apart from revealing a blood tramadol concentration well above the normal therapeutic range. This is the first report of fatal tramadol ingestion occurring in a therapeutic setting and also the first tramadol-related death where the mechanism was liver failure. PMID- 12850424 TI - Naphazoline intoxication in a child-a clinical and forensic toxicological case. AB - The imidazoline derivative naphazoline, an alpha(2)-adrenergic agonist, is used as non-prescription eye and nasal preparation because of its vasoconstrictive and decongestive properties. Especially in children, overdose and/or systemic side effects due to absorption can quickly cause severe central nervous system depression and cardiovascular adverse effects. In a 7-year-old boy was diagnosed a naphazoline intoxication by toxicological analysis. The case was also of forensic interest, because the naphazoline mixture was prepared in a pharmacy in a concentration 80 times above the adequate dosage for children. In general, physicians, pharmacists and the public should be educated about the toxicity of over-the-counter preparations. PMID- 12850425 TI - Molecular dissection of neuromuscular junction formation. AB - The muscle-specific kinase (MuSK) is the key component that mediates the synapse inducing role of motoneuron-derived agrin at the neuromuscular junction. Recent reverse-genetics approaches have shed new light on the events triggered inside myotubes by activation of this kinase. Mice in which most of the intracellular domain of MuSK is replaced by a related kinase are viable. Analysis of the imperfect postsynaptic specializations of these mice has provided new insights into the complex postsynaptic differentiation process. PMID- 12850426 TI - Prion protein trafficking and the development of neurodegeneration. AB - The prion protein (PrP) is involved in causing a group of diverse transmissible, heritable and sporadically occurring neurodegenerative diseases. Although the identity, nature and replication of the transmissible agent have been intensely studied for decades, the cellular events underlying neuronal dysfunction and death have received comparatively little attention. Recent studies examining the occurrence and consequences of inappropriate cytoplasmic expression of the normally cell-surface PrP underscore an emerging role for PrP trafficking in prion disease pathogenesis. PMID- 12850427 TI - Astrocyte-mediated control of cerebral microcirculation. AB - Characterization of astrocyte Ca2+ dynamics has been a topic of considerable emphasis for more than a decade. Only recently, however, has the physiological significance of astrocyte Ca2+ signaling started to become clear. Several studies have shown that astrocyte Ca2+ levels become elevated in response to neuronal input and that this, in turn, influences synaptic activity. A novel function of astrocyte Ca2+ signaling has been described by Zonta et al., whereby neuron induced astrocyte Ca2+ elevations can lead to secretion of vasodilatory substances from perivascular astrocyte endfeet, resulting in improved local blood flow. This finding represents a breakthrough in our knowledge both of astrocyte function and of the mechanism of activity-dependent cerebral blood flow regulation. PMID- 12850430 TI - Sound localization and delay lines--do mammals fit the model? AB - The current dominant model of binaural sound localization proposes that the lateral position of a sound source is determined by the position of maximal activation within an array of binaural coincidence-detector neurons that are tuned to different interaural time differences (ITDs). The tuning of a neuron for an ITD is determined by the difference in axonal conduction delay from each ear- the so-called "delay line" hypothesis. Although studies in birds appear to support this model, recent evidence from mammals suggests that the model does not provide accurate descriptions of how ITDs are encoded in the mammalian auditory brainstem or of how ITD-sensitive neurons contribute to mammalian sound localization. PMID- 12850429 TI - The rotenone model of Parkinson's disease. PMID- 12850431 TI - The stem-cell menagerie. AB - The numbers, types and locations of stem cells in the nervous system have been the subject of much discussion. This review summarizes data on the types of stem cell present at different stages of development and in the adult brain, and the markers suggested to distinguish between the various possibilities that have been reported. We present evidence that more than one class of stem cell is present in the developing and adult nervous systems, and that it might be possible to distinguish between stem-cell populations and to localize the cell of origin of a particular neurosphere, based on markers that persist in culture and by using universal stem-cell markers prospectively to identify stem cells in vivo. PMID- 12850432 TI - Structure-stability-function relationships of dendritic spines. AB - Dendritic spines, which receive most of the excitatory synaptic input in the cerebral cortex, are heterogeneous with regard to their structure, stability and function. Spines with large heads are stable, express large numbers of AMPA-type glutamate receptors, and contribute to strong synaptic connections. By contrast, spines with small heads are motile and unstable and contribute to weak or silent synaptic connections. Their structure-stability-function relationships suggest that large and small spines are "memory spines" and "learning spines", respectively. Given that turnover of glutamate receptors is rapid, spine structure and the underlying organization of the actin cytoskeleton are likely to be major determinants of fast synaptic transmission and, therefore, are likely to provide a physical basis for memory in cortical neuronal networks. Characterization of supramolecular complexes responsible for synaptic memory and learning is key to the understanding of brain function and disease. PMID- 12850433 TI - Molecular basis of plasticity in the visual cortex. AB - Sensory experience is known to shape the maturation of cortical circuits during development. A paradigmatic example is the effect of monocular deprivation on ocular dominance of visual cortical neurons. Although visual cortical plasticity has been widely studied since its initial discovery by Hubel and Wiesel >40 years ago, the description of the underlying molecular mechanisms has lagged behind. Several new findings are now beginning to close this gap. Recent data deepen our knowledge of the factors involved in the intercellular communication and intracellular signaling that mediate experience-dependent plasticity in the developing visual cortex. In addition, new findings suggest a role for the extracellular matrix in inhibition of ocular-dominance plasticity in the adult visual cortex. PMID- 12850434 TI - New directions in retinal research. AB - Direction-selective retinal ganglion cells (DSGCs) respond to image motion in a "preferred" direction but not the opposite "null" direction. Extracellular spike recordings from rabbit DSGCs suggested that the key mechanism underlying the directional responses is spatially offset inhibition projecting in the null direction. Recent patch-clamp recordings have shown that this inhibition, which acts directly on the DSGC, is already direction selective. Dual recordings established that the inhibition arises from starburst amacrine cells (SBACs) located on the null side of the DSGC but not from those on the preferred side. Thus, for each radially symmetric SBAC, processes pointing in different directions would provide the null-direction inhibition to subtypes of DSGCs with different preferred directions. Ca2+ imaging revealed that the SBAC terminal processes respond more strongly to image motion away from the soma than towards the soma, therefore accounting for the direction selectivity of the inhibitory input to the DSGCs. PMID- 12850435 TI - Modeling CNS neurodegeneration by overexpression of disease-causing proteins using viral vectors. AB - Defective handling of proteins is a central feature of major neurodegenerative diseases. The discovery that neuronal dysfunction or degeneration can be caused by mutations in single cellular proteins has given new opportunities to model the underlying disease processes by genetic modification of cells in vitro or by generation of transgenic animals carrying the disease-causing gene. Recent developments in recombinant viral-vector technology have opened up an interesting alternative possibility, based on direct gene transfer to selected subregions or subsets of neurons in the brain. Using the highly efficient adeno-associated virus or lentivirus vectors, recent reports have shown that overexpression of mutated human huntingtin or alpha-synuclein in neurons in the striatum or substantia nigra induces progressive neuropathology and neurodegeneration, similar to that seen in Huntington's and Parkinson's diseases. Targeted overexpression of disease-causing genes by recombinant viral vectors provides a new and highly flexible approach for in vivo modeling of neurodegenerative diseases, not only in mice and rats but also in primates. PMID- 12850436 TI - Special issue of TiG. PMID- 12850437 TI - Evolution of duplicate genes versus genetic robustness against null mutations. PMID- 12850438 TI - Evolutionary diversification of mitochondrial proteomes: implications for human disease. PMID- 12850439 TI - Human housekeeping genes are compact. PMID- 12850440 TI - Strand misalignments lead to quasipalindrome correction. PMID- 12850441 TI - ITS2 is a double-edged tool for eukaryote evolutionary comparisons. PMID- 12850442 TI - Ciona intestinalis: an emerging model for whole-genome analyses. PMID- 12850443 TI - Control of protein degradation by E3 ubiquitin ligases in Drosophila eye development. PMID- 12850444 TI - Recent highlights of the China Rice Functional Genomics Program. PMID- 12850445 TI - Central synapse and neuromuscular junction: same players, different roles. PMID- 12850446 TI - Cytokinesis in fission yeast: a story of rings, rafts and walls. PMID- 12850447 TI - Methyl jasmonate as a vital substance in plants. AB - The plant floral scent methyl jasmonate (MeJA) has been identified as a vital cellular regulator that mediates diverse developmental processes and defense responses against biotic and abiotic stresses. The pleiotropic effects of MeJA have raised numerous questions about its regulation for biogenesis and mode of action. Characterization of the gene encoding jasmonic acid carboxyl methyltransferase has provided basic information on the role(s) of this phytohormone in gene-activation control and systemic long-distance signaling. Recent approaches using functional genomics and bioinformatics have identified a whole set of MeJA-responsive genes, and provide insights into how plants use volatile signals to withstand diverse and variable environments. PMID- 12850448 TI - A biomechanical model for analysis of muscle force, power output and lower jaw motion in fishes. AB - Fish skulls are complex kinetic systems with movable components that are powered by muscles. Cranial muscles for jaw closing pull the mandible around a point of rotation at the jaw joint using a third-order lever mechanism. The present study develops a lever model for the jaw of fishes that uses muscle design and the Hill equation for nonlinear length-tension properties of muscle to calculate dynamic power output. The model uses morphometric data on skeletal dimensions and muscle proportions in order to predict behavior and force transmission mediated by lever action. The computer model calculates a range of dynamic parameters of jaw function including muscle force, torque, effective mechanical advantage, jaw velocity, bite duration, bite force, work and power. A complete list of required morphometrics is presented and a software program (MandibLever 2.0) is available for implementing lever analysis. Results show that simulations yield kinematics and timing profiles similar to actual fish feeding events. Simulation of muscle properties shows that mandibles reach their peak velocity near the start of jaw closing, peak force at the end of jaw closing, and peak power output at about 25% of the closing cycle time. Adductor jaw muscles with different mechanical designs must have different contractile properties and/or different muscle activity patterns to coordinate jaw closing. The effective mechanical advantage calculated by the model is considerably lower than the mechanical advantage estimated from morphological lever ratios, suggesting that previous studies of morphological lever ratios have overestimated force and underestimated velocity transmission to the mandible. A biomechanical model of jaw closing can be used to interpret the mechanics of a wide range of jaw mechanisms and will enable studies of the functional results of developmental and evolutionary changes in skull morphology and physiology. PMID- 12850449 TI - Oscillations in cyclical neutropenia: new evidence based on mathematical modeling. AB - We present a dynamical model of the production and regulation of circulating blood neutrophil number. This model is derived from physiologically relevant features of the hematopoietic system, and is analysed using both analytic and numerical methods. Supercritical Hopf bifurcations and saddle-node bifurcations of limit cycles are shown to exist. We make the estimation of kinetic parameters for dogs and then apply the model to cyclical neutropenia (CN) in the grey collie, a rare disorder in which oscillations in all blood cell counts are found. We conclude that the major cause of the oscillations in CN is an increased rate of apoptosis of neutrophil precursors which leads to a destabilization of the hematopoietic stem cell compartment. PMID- 12850450 TI - Structural determinants of the rate of protein folding. AB - To understand the mechanism of protein folding and to assist rational design of fast-folding, non-aggregating and stable artificial enzymes, it is essential to determine the structural parameters which govern the rate constants of folding, kf. It has been found that -logkf is a linear function of the so-called chain topology parameter (CTP) within the range of 10(-1)s(-1)< or = kf < or =10(8)s( 1). The correlation between -logkf and CTP is much improved than using previously published contact order (CO) method. It has been further suggested that short sequence separations may be preferred for the establishment of stable interactions for the design of novel artificial enzymes and the modification of slow-folding proteins with aggregating intermediates. PMID- 12850451 TI - Does a structural bridge exist between the DNA and the specialized cytoplasmic organelles during the early part of their development? A mechanism for the positioning of flagella and possibly other cytoplasmic organelles. AB - Cell differentiation involves the development of a new cytoplasm containing a set of specialized organelles such as cilia and flagella which are placed in the cell with a predetermined orientation. Arguments are put forward to show that the orientation of the flagellar apparatus could be brought about by a macromolecular structural bridge between the nucleoid and the assembling flagellar apparatus, the orientation being determined by the spatial geometry inherent in the folding of the DNA. An analysis of differentiation in unicelled eukaryotes suggests that the same basic mechanism of a structural bridge could also apply to the orientation of their cilia and flagella and perhaps may have a more general application in the positioning of cytoplasmic organelles. PMID- 12850452 TI - Organization of the cytokeratin network in an epithelial cell. AB - The cytoskeleton is a dynamic three-dimensional structure mainly located in the cytoplasm. It is involved in many cell functions such as mechanical signal transduction and maintenance of cell integrity. Among the three cytoskeletal components, intermediate filaments (the cytokeratin in epithelial cells) are the best candidates for this mechanical role. A model of the establishment of the cytokeratin network of an epithelial cell is proposed to study the dependence of its structural organization on extracellular mechanical environment. To implicitly describe the latter and its effects on the intracellular domain, we use mechanically regulated protein synthesis. Our model is a hybrid of a partial differential equation of parabolic type, governing the evolution of the concentration of cytokeratin, and a set of stochastic differential equations describing the dynamics of filaments. Each filament is described by a stochastic differential equation that reflects both the local interactions with the environment and the non-local interactions via the past history of the filament. A three-dimensional simulation model is derived from this mathematical model. This simulation model is then used to obtain examples of cytokeratin network architectures under given mechanical conditions, and to study the influence of several parameters. PMID- 12850453 TI - A selfish origin for recombination. AB - Recent findings of molecular biology show that recombination is initiated by interactions between homologous chromosomes and that an allele can induce the initiation of recombination on the homolog. Since gene conversion at the site of initiation is strong enough to promote the transmission of that allele, recombination may be a way for a self-promoting element to spread, even if it gives no advantage to the individual or to the population. I develop a simple model and discuss available molecular evidence in support of this hypothesis. A consequent argument is that with asexual reproduction the evolution of recombination leads to an intragenomic conflict, and a possible outcome of this conflict may be the origin of sexual reproduction. PMID- 12850454 TI - Modelling of Dictyostelium discoideum slug migration. AB - The development of most multicellular organisms involves differential movement of cells resulting in the formation of tissues. The principles governing these movements are poorly understood. One exception is the formation of the slug in the social amoebae Dictyostelium discoideum. The slug forms by the chemotactic aggregation of up to 10(5) starving cells, it is motile and migrates in response to light and temperature gradients to the surface of the soil to form a fruiting body consisting of a stalk supporting a spore head. Slug migration and behaviour result from coordinated chemotactic movement of the individual cells in the slug. Waves of a chemoattractant, most likely cAMP, are periodically initiated in the tip of the slug and propagate towards the back of the slug resulting in periodic forward movement of individual cells as well as the whole slug. Here we develop a model to investigate how wave propagation and cell movement interacts to result in migration and shape changes of the slug. The slug tissue is modelled as an incompressible liquid, in which waves of chemoattractant are generated in an excitable manner. The liquid is "active", i.e. it is able to generate body forces in response to the gradients of the chemoattractant. These forces lead to the formation of flows (representing chemotactically moving cells) and result in slug movement and shape changes. The model provides a theoretical framework for the understanding of the interactions between cell-cell signalling and cell movement, which govern slug behaviour and tissue morphogenesis. PMID- 12850455 TI - Baseline criteria and the evolution of hosts and parasites: D0, R0 and competition for resources between strains. AB - Our understanding of the evolution of diseases has been greatly aided by the use of baseline criteria. Here we examine the theoretical and biological relationships of the well known baseline criteria for the evolution of disease (R0) and the recently introduced corresponding criterion for the evolution of resistance in hosts (D0). We show that there is a formal theoretical equivalence between the two criteria and discuss the characteristics of seperability that determine whether the criteria define the course of evolution. These theoretical determinants correspond biologically to whether strains compete for resources or not. We discuss the biological application of the criteria and argue that D0 may be less widely applicable than R0, but does determine the evolution of resistance in populations with fixed carrying capacities. PMID- 12850456 TI - The relationship between neuronal calcium concentration and firing rate during stochastic synaptic inputs. AB - We propose a novel, nonlinear theory about reading neuronal information using intracellular calcium concentrations, which includes the linear theory already developed in the literature as a special case. The theory is numerically confirmed using the Pinsky-Rinzel and integrate-and-fire models with constant rate Poisson inputs. Applying the theory to models with non-constant inputs, we find that there is a time lag equal to the calcium buffering time constant between the instantaneous firing rate and the firing rate estimated using calcium concentrations. PMID- 12850457 TI - Testing fundamental evolutionary hypotheses. AB - Sober and Steel (J. Theor. Biol. 218, 395-408) give important limits on the use of current models with sequence data for studying ancient aspects of evolution; but they go too far in suggesting that several fundamental aspects of evolutionary theory cannot be tested in a normal scientific manner. To the contrary, we show examples of how some alternatives to the theory of descent can be formulated in such a way that they lead to predictions that can be evaluated (and rejected). The critical factor is a logical formulation of the alternatives, even though not all possible alternatives can be tested simultaneously. Similarly, some of the limits using DNA sequence data can be overcome by other types of sequence derived characters. The uniqueness (or not) of the origin of life, though still difficult, is similarly amenable to the testing of alternative hypotheses. PMID- 12850458 TI - The choice of cell lineages during the in vitro differentiation of mammalian embryonic stem cells. PMID- 12850459 TI - Mouse model of heterotopic aortic arch transplantation. AB - BACKGROUND: Syngeneic heterotopic transplantation of segments of descending thoracic aortas containing atherosclerotic lesions from hypercholesterolemic mice into normocholesterolemic recipients has been useful for studies on plaque regression and stabilization. Because lesion development is more rapid and exuberant in the aortic arch, a technique of transplantation of the mouse aortic arch was developed. MATERIALS AND METHODS: C57BL/6, apoE-deficient (apoE-/-) (hypercholesterolemic) mice were fed a Western diet for 22 weeks and used as donors of aortic-arch segments containing atherosclerotic lesions. Twenty syngeneic transplants were performed on age-matched wild-type (normocholesterolemic) mice. Aortic arches containing atherosclerotic lesions were implanted on the abdominal aorta of recipient mice by end-to-side microsurgical anastomosis. Two weeks after transplantation, grafts were noninvasively imaged in vivo by magnetic resonance (MR) microscopy. Grafts harvested four weeks after transplantation were submitted for histological examination. RESULTS: All recipients survived the entire follow-up period (1 month) without complications. Duration of recipient procedure ranged from 90 to 120 (mean, 105) min; aortic clamping time varied from 45 to 60 min. In vivo MR microscopy demonstrated patency of the grafts and wall thickening that corresponded to the preexisting atherosclerotic lesions. Histology confirmed patency and atherosclerotic thickening of the grafts, and showed no evidence of acute tissue damage. CONCLUSIONS: Syngeneic transplantation of the aortic arch in mice represents a useful alternative model for studies on morphology, imaging, and mechanisms of atherosclerosis. The curvature of the aortic arch is preserved after implantation onto the abdominal aorta, providing clear landmarks for noninvasive assessment using MR. PMID- 12850460 TI - Focal adhesions and associated proteins in medullary thyroid carcinoma cells. AB - BACKGROUND: In medullary thyroid carcinoma (MTC), mutations in the RET protooncogene lead to oncogenic transformation. RET activation in other cell types has been shown to cause phosphorylation of the focal adhesion-associated proteins focal adhesion kinase (FAK), paxillin, and p130(CAS). We hypothesized that adhesion-dependent signaling might be deranged in MTC cells. METHODS: Indirect immunofluorescence was used to label beta(1) integrin, FAK, paxillin, and p130CAS. Rhodamine-labeled phalloidin was used to visualize actin microfilaments. Phosphorylated protein was detected by immunoprecipitation followed by Western blotting for phosphotyrosine. MTC cell invasiveness was quantified using a modified Boyden chamber assay. RESULTS: Clustering of beta(1) integrin, FAK, paxillin, and p130(CAS) into focal adhesions were not detected in MTC cells under any conditions, although clustering was seen as expected in control HeLa cells. Despite this failure, FAK, paxillin and p130(CAS) were all found to be phosphorylated. Actin microfilaments were generally not seen although in a few cells, small, poorly formed microfilaments could be detected. MTC cells invaded poorly as compared with highly invasive cell lines. However a clear difference was noted between invasiveness on growth factor depleted Matrigel and regular Matrigel. CONCLUSIONS: In MTC cells, focal adhesions are not seen in response to interaction with extracellular matrix. Consistent with this failure, actin microfilaments are absent or poorly formed and invasion is weak. Despite the absence of focal adhesions, focal adhesion proteins remain phosphorylated, even though in normal cells their signaling activity is dependent on focal adhesion formation. Deranged adhesion-dependent signaling may contribute to MTC pathogenesis. PMID- 12850461 TI - Metalloproteinase inhibition reduces lung injury and improves survival after cecal ligation and puncture in rats. AB - BACKGROUND: Neutrophil activation with concomitant matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) release has been implicated in the development of sepsis-induced acute lung injury. We hypothesized that COL-3, a chemically modified tetracycline known to inhibit MMP-2 and MMP-9, would reduce lung injury and improve survival in rats following cecal ligation and puncture (CLP). METHODS: Sprague-Dawley rats were separated into five groups: 1) sham CLP+ carboxymethylcellulose (CMC; vehicle for COL-3, n = 6); 2) sham CLP + COL-3 (n = 6); 3) CLP + CMC (n = 10); 4) CLP + single-dose (SD) COL-3 administered concomitant with CLP (n = 9); and 5) CLP + multiple-dose (MD) COL-3 administered concomitant with CLP and at 24 h after CLP (n = 15). Rats were sacrificed at 168 h (7 days) or immediately after death, with survival defined as hours after CLP. Histological lung assessment was made based on neutrophil infiltration, alveolar wall thickening, and intraalveolar edema fluid. Lung MMP-2 and MMP-9 levels were assessed by immunohistochemistry. MMP-2 and MMP-9 levels were correlated with survival by simple regression analysis. RESULTS: The mortality of rats in the cecal ligation and puncture without treatment group (CLP + CMC) was 70% at 168 h. A single dose of COL-3 in the CLP + COL-3 (SD) group significantly reduced mortality to 54%. Furthermore, with a repeat dose of COL-3 at 24 h after CLP, mortality was significantly reduced to 33%. Pathologic lung changes seen histologically in the CLP + CMC group were significantly reduced by COL-3. A significant reduction in lung tissue levels of MMP-2 and MMP-9 was noted in both groups treated with COL-3. Reduction of MMP-2 and MMP-9 levels correlated with improved survival. CONCLUSION: Inhibition of MMP-2 and MMP-9 by COL-3 in a clinically relevant model of sepsis-induced acute lung injury reduces pulmonary injury and improves survival in a dose-dependent fashion. Our results suggest that prophylactic treatment with COL-3 in high-risk patients may reduce the morbidity and mortality associated with sepsis-induced acute respiratory distress syndrome. PMID- 12850462 TI - Loss of p27(Kip1) accelerates DNA replication after partial hepatectomy in mice. AB - BACKGROUND: Liver regeneration after partial hepatectomy (PH) is accomplished by a synchronous replication of hepatocytes. Both positive and negative regulators of cyclin-dependent protein kinase (Cdk) have been implicated in hepatocyte proliferation, but their specific roles in vivo remain to be clarified. To investigate the specific role of p27(Kip1), a member of the Cip/Kip family of Cdk inhibitors, in cell-cycle regulation during liver regeneration, p27-knockout mice were studied after PH. MATERIALS AND METHODS: Under ether anesthesia, mice were subjected to 70% PH. Animals were sacrificed at intervals after the surgery, and the remnant liver was harvested and analyzed. RESULTS: In p27-deficient mice, the timing of DNA synthesis was significantly accelerated with a perturbation in the ordered distribution of proliferating cells in the hepatic lobule. p27 deficiency, however, did not affect the whole population of cycling cells, the number of apoptotic cells, or liver injury and mortality after PH. CONCLUSION: These data provide in vivo evidence that p27 functions as a brake in the "start" of the hepatocyte cell cycle, thereby coordinating temporally and spatially the onset of DNA synthesis of hepatocytes within the hepatic lobules. PMID- 12850463 TI - The combination of polymicrobial sepsis and endotoxin results in an inflammatory process that could not be predicted from the independent insults. AB - BACKGROUND: The variable clinical profile observed in critically ill patients is the result of multiple factors. Genetic determinants have recently been shown as confounding factors in the response to injury. However, other elements, such as the environment and the type of injury, could modify this response. The objective of this investigation was to study the effect of combining insults and different genetic backgrounds on the inflammatory response. MATERIALS: Male mice, C57BL/6J (B6) and A/J, were randomized to undergo cecal ligation and single puncture (CLP) or sham operation (SOP). After 24 h of recovery, mice were randomized again into two groups, one group was injected with bacterial lipopolysaccharide (LPS; 15 mg/kg) and the other was injected with normal saline (NS). An additional experimental group included mice that were not operated (NOP) and injected with LPS. Mice were evaluated by plasma cytokine content. RESULTS: The combination of insults resulted in an apparent additive effect for some cytokines, such as interleukin (IL) 6. In contrast, tumor necrosis factor alpha (TNF-alpha) was considerably lower in the combined injury group with respect to injection of LPS alone. There was no relevant difference in IL-10 levels between any group, except that its decay was slower in the CLP + LPS group. Overall, cytokine levels were different between B6 and A/J mice indicating a genetic contribution. CONCLUSIONS: These results indicate that the response to stress is the combination of the type of injury and the genetic background of the subject. These observations also illustrate the difficulty in predicting the inflammatory response and underlying mechanism based on cytokine plasma levels. PMID- 12850464 TI - Bacterial translocation after cirrhotic liver resection: a clinical investigation of 181 patients. AB - BACKGROUND: Cirrhotic patients are usually associated with a high susceptibility to infection. Although bacterial translocation from gut mucosa to mesenteric lymph node (MLN) and systemic circulation is a well-known phenomenon after hepatectomy, its role in cirrhotic patients remains unclear. MATERIALS AND METHODS: MLN was harvested for bacterial culture before and after liver resection in 181 cirrhotic patients. The characteristics and postoperative courses of patients with positive and negative bacterial culture for MLN after hepatectomy were compared. Postoperative systemic antibiotics were administered if infectious complications occurred. RESULTS: No bacteria were cultured in MLN before hepatectomy. Bacterial translocation (BT) to MLN after hepatectomy occurred in 36 patients (BT group). After multivariate analysis, intraoperative blood transfusion was the only independent factor that influenced bacterial translocation rates after cirrhotic liver resection. BT group patients also had higher infectious and overall complication rates, with a longer postoperative hospital stay. Among the cultured bacteriae from infected sites in BT group patients with infectious complications, only 2 patients (12.5%) had totally different bacterial species to those cultured from MLNs. CONCLUSIONS: Bacterial translocation more often occurred after liver resection in cirrhotic patients who received intraoperative blood transfusion. Such patients had higher postoperative infectious and overall complication rates. Thus, avoidance of intraoperative blood transfusion is mandatory for cirrhotic liver resection. PMID- 12850465 TI - Cytokine signatures in atherosclerotic claudicants. AB - BACKGROUND: Iron accumulation and inflammation may affect atherosclerosis. This study intended to define a cytokine signature in atherosclerotic claudicants and to determine whether reduction in serum ferritin by phlebotomy influenced this pattern. METHODS: Ninety-one subjects with peripheral vascular disease (PVD; mean age, 67 years) were recruited from the VA Cooperative Iron and Atherosclerosis Study (FeAST) testing the hypothesis that ferritin reduction to 25 ng/ml may ameliorate atherosclerosis. Cytokines TNF-a, IL-2, IL-6, and IL-10 were analyzed by enzyme amplified sensitivity assay (EASIA). Fasting iron and cholesterol panels, complete blood count, C-reactive protein (CRP), uric acid, fibrinogen, glucose, and hemoglobin A1c levels were also quantified. Values were compared with "healthy" controls (n = 21; mean age, 56 years). After randomization of PVD to phlebotomy (intervention group [IG], n = 44) or control (nonintervention group [NG], n = 47), analyses were compared at 6 and 12 months using t test, Wilcoxon rank sum test, chi-square, and robust MM regression. FINDINGS: Age, glucose, and hemoglobin A1c were higher in PVD compared with healthy controls (P < 0.01), whereas serum iron (P < 0.01) and percentage of transferrin saturation (P < 0.05) were lower. Tumor necrosis factor-alpha (TNF-alpha; P < 0.05), IL-6 (P < 0.01), and CRP (P < 0.05) levels were higher in the PVD group, whereas IL-10 was lower (P < 0.01). At 6 months post phlebotomy, ferritin levels were reduced (P < 0.01), although ferritin levels were reduced less in smokers. IL-6 and fibrinogen, CRP and ferritin levels correlated positively. At 6 and 12 months, subjects with TNF alpha (n= 15) and IL-6 (n = 10) levels in the upper 25th percentile were reduced by phlebotomy. INTERPRETATION: An inflammatory cytokine signature exists in atherosclerosis. Elevated levels of TNF-alpha and IL-6, reportedly associated with recurrent and future myocardial infarction, were reduced by phlebotomy. The utility of the iron/inflammatory hypotheses will ultimately relate to clinical outcomes obtained prospectively by the FeAST trial. PMID- 12850466 TI - Effect of glutamine on glutathione, IGF-I, and TGF-beta 1. AB - BACKGROUND: Our previous results have showed that oral glutamine (GLN) supplementation decreased carcinogenesis in 7,12-dimethylbenz[a]antracene (DMBA) breast cancer model. We also have found that GLN raises blood glutathione (GSH) levels in an implantable breast cancer model. The process of tumor growth was accompanied by depressed GSH production and increased levels of insulin-like growth factor-I (IGF-I) and transforming growth factor beta1 (TGF-beta 1). GSH is counter-regulatory to IGF-I. We therefore hypothesized that in DMBA model of breast cancer, the increased GSH levels seen with oral GLN would be associated with lowered levels of IGF-I &TGF-beta(1). METHODS: Time-dated pubertal Sprague Dawley rats were gavaged at time 0 with 1 g/kg/day glutamine (GLN) (n = 18), isonitrogenous Freamine (FA) (n = 18), or water (H(2)O) (n = 18). Rats were further randomized on day 7 to 100 mg/kg DMBA or oil. After 14 days, the animals were sacrificed and blood GSH, IGF-1, TGF-beta 1, breast tissue, and gut mucosa GSH levels were measured. RESULTS: Oral GLN increased significantly blood, breast tissue, and gut mucosa levels of GSH in both DMBA and control groups in comparison with the control groups not treated with GLN. At the same time, the levels of blood IGF-I and TGF-beta 1 decreased significantly in both DMBA-treated and control groups. DMBA did not significantly affect any of these levels. CONCLUSIONS ;Oral GLN increased GSH levels and lowered IGF-I and TGF-beta 1 in a range that is considered clinically significant. However, the effect of GLN in maintaining normal gut GSH production in the presence of DMBA was much more significant. Inconsistent with our hypothesis, reduction in IGF and TGF-beta 1 levels did not correlate with DMBA's effect on gut GSH production. PMID- 12850467 TI - Xenogeneic direct hemoperfusion using whole swine liver for liver failure in dogs. AB - BACKGROUND: We developed a new method of xenogeneic direct hemoperfusion of a bioartificial liver support system consisting of a leukocyte-adsorbent column, an immunoglobulin-adsorbent column, and the substitute unit for hepatic function. By this method, we performed xenogeneic direct hemoperfusion experiment using resected whole swine liver for treatment of a canine liver failure model, and compared the contribution of each adsorbent column both by blood analysis and from the histological point of view. MATERIALS AND METHODS: Canine liver failure model was produced by portocaval shunting and ligating the entire hepatoduodenal ligament. The xenogeneic direct hemoperfusion system was constructed using a roller pump, a leukocyte-adsorbent column, an immunoglobulin-adsorbent column, a combined device of oxygenator and warmer, the resected whole swine liver accommodated in a chamber, and a dissolved oxygen meter through which canine whole blood leaving the external jugular vein circulated in this order. RESULTS: Xenogeneic direct hemoperfusion was successfully performed for 3 h without hyperacute rejection occurring. Adequate ammonia detoxification and bile juice secretion were exhibited, and no findings of hepatocyte destruction by immunological cells and proteins were detected. Blood data showed that the immunoglobulin adsorbent were more effective than the leukocyte adsorbent in avoiding hyperacute rejection. This result indicates that hyperacute rejection has a closer relation to humoral immune responses, especially regarding removal of complements than to cellular immune responses. CONCLUSIONS: We successfully performed xenogeneic direct hemoperfusion of the whole swine liver without hyperacute rejection using our method. PMID- 12850468 TI - Attenuation of ischemia/reperfusion injury by N-acetylcysteine in a rat hind limb model. AB - BACKGROUND: Ischemia/reperfusion is a complex set of events with severe pathologic consequences. Reperfusion initiates both the local and systemic damage in part through rapid oxygen generation. N-acetylcysteine (NAC) is a scavenger of free radical species, inhibits neutrophil accumulation, acts as a vasodilator and also improves microcirculation. In present study, we examined the protective effect of NAC in a rat hind limb ischemia/ reperfusion model. Dimethyl-sulfoxide (DMSO), a well-known antioxidant was also tested for comparison. MATERIALS AND METHODS: Ischemia was induced for 4 h by vascular clamping and followed by 1 h of reperfusion. Muscle injury was evaluated in 3 groups as a saline group (control), DMSO group, and NAC group. Plasma levels of creatine kinase, lactate dehydrogenase, thiobarbituric acid reactive substances (TBARS), and blood HCO(3), as well as muscle tissue TBARS, were measured at the end of reperfusion. Muscle tissue samples were taken for histological evaluation. RESULTS: DMSO and NAC group showed significant amelioration of plasma CPK (P < 0.05, P < 0.05), plasma TBARS (P < 0.05, P < 0.05), and muscle tissue TBARS (P < 0.05, P < 0.05) compared with the control group. Similarly, neutrophil infiltration in DMSO and NAC groups were significantly less prominent than the control group (P < 0.01, P < 0.01). CONCLUSIONS: These results show that NAC improved effectively ischemia reperfusion injury in a rat hind limb model. PMID- 12850469 TI - Mitochondrial permeability transition and cytochrome c release in ischemia reperfusion injury of the rat liver. AB - BACKGROUND: We investigated whether ischemia-reperfusion causes activation of caspases and whether this activation is related to cytochrome c release from the mitochondria into the cytosol as a result of the mitochondrial inner membrane permeability transition. MATERIALS AND METHODS: Rats were subjected to 30 min to 120 min of hepatic ischemia followed by 6 h of reperfusion. Cyclosporin A or ruthenium red (inhibitors of the mitochondrial inner membrane permeability transition) was given intravenously at 60 and 30 min before ischemia, respectively. RESULTS: Reperfusion after ischemia caused the release of liver enzymes accompanied by mitochondrial membrane depolarization, DNA fragmentation, and translocation of cytochrome c from the mitochondria into the cytosol. Accumulation of cytochrome c in the cytosol and activation of caspase-3-like protease was already detected during ischemia and before reperfusion. Pretreatment with cyclosporin A or ruthenium red significantly ameliorated the loss of the mitochondrial membrane potential, the increase of plasma membrane permeability, the cytosolic accumulation of cytochrome c, DNA fragmentation, and caspase-3-like protease activation. CONCLUSIONS: The mitochondrial inner membrane permeability transition occurs during ischemia and/or after reperfusion, resulting in translocation of cytochrome c and activation of caspases. PMID- 12850470 TI - Oxidative stress in patients undergoing cardiac surgery: comparative study of revascularization and valve replacement procedures. AB - OBJECTIVE: The aim of this study was to evaluate the time course of oxidative stress markers in plasma and erythrocyte from patients undergoing open heart surgery with cardiopulmonary bypass (CPB) and to examine whether the type of surgical technique used (valve replacement or coronary revascularization) produces any differences in these makers. PATIENTS AND METHODS: Twenty-two patients undergoing cardiac surgery with CPB were divided in 2 groups (valve replacement or coronary revascularization). We took 5 blood samples at different times during cardiac surgery and analyzed thiobarbituric acid reactive substances (TBARS), alpha-tocopherol, coenzyme Q, and retinol in plasma and TBARS (baseline levels and induced by Fe(2+)-ascorbate oxidation), alpha-tocopherol, coenzyme Q and catalase, superoxide dismutase, and glutathione peroxidase activity in erythrocyte. RESULTS: Plasma alpha-tocopherol content decreased after starting CPB in both groups. In contrast, in erythrocytes there was an increase in the activity or concentration of all of the antioxidants. Erythrocyte TBARS contents, both baseline levels and induced levels, were higher in coronary revascularization group. CONCLUSION: Although both groups suffered an increase in oxidative stress after CPB, this increase was higher in coronary revascularization group and therefore the possibility of post-CPB complications could be more severe in this group. As the groups followed a different pattern of antioxidant response, a different therapeutic approach may be required for each. PMID- 12850471 TI - Intermittent hepatic ischemia-reperfusion minimizes liver metastasis in rats. AB - BACKGROUND: Surgical stresses, including hepatic ischemia-reperfusion (I/R), promote cancer growth and metastasis. We have reported that continuous hepatic I/R increases liver damage and promoted liver metastasis from colon cancer, whereas intermittent I/R causes less liver damage. We therefore examined whether intermittent I/R could reduce liver metastasis in a rat model. MATERIALS AND METHODS: Adult male Fischer rats was divided between three groups: group A (control), which received laparotomy for 120 min with no liver ischemia; group B (continuous I/R), which received 60 min of 70% partial liver ischemia followed by 60 min of reperfusion; and group C (intermittent I/R), which received 15 min of 70% ischemia and 15 min of reperfusion, repeated four times. Just before closing the abdomen, all animals were inoculated intrasplenically with rat colon adenocarcinoma cells (RCN-H4). Tumor nodules on the liver surface were counted 3 weeks later. In addition, expression of E-selectin mRNA in liver was examined at 1, 3, and 6 h after completing I/R by a reverse transcription-polymerase chain reaction. RESULTS: Continuous I/R (B) greatly promoted liver metastasis in both ischemic and nonischemic liver lobes, whereas intermittent I/R (C) showed significantly fewer metastasis than group B in both lobes. Significantly less E selectin mRNA was expressed in group C than in group B. CONCLUSIONS: Intermittent I/R limits expression of E-selectin mRNA and liver metastasis. Intermittent hepatic I/R is less stressful than continuous I/R, minimizing liver metastasis by colon cancer cells through avoidance of E-selectin up-regulation. PMID- 12850472 TI - Smooth muscle cell migration stimulated by interleukin 6 is associated with cytoskeletal reorganization. AB - BACKGROUND: Interleukin 6 (IL-6) is elevated in the arterial wall in atherosclerosis and restenosis after angioplasty. An important contributor to these pathologies is migration of vascular smooth muscle cells (VSMC), which is often associated with cytoskeletal reorganization initiated by growth factors and chemokines. We recently reported that IL-6 stimulated migration of VSMC. Here, we examined the cytoskeleton of VSMC and cytoskeletal associated proteins to determine potential mechanisms associated with IL-6 induced migration. MATERIALS AND METHODS: Studies were performed in VSMC cultured from rat aortas. RESULTS: IL 6 significantly stimulated VSMC migration. IL-6 induced actin polymerization, and tyrosine phosphorylation of focal adhesion-associated cytoskeletal proteins including focal adhesion kinase (FAK) and paxillin. Cytochalasin D, an inhibitor of actin polymerization, blocked phosphorylation of FAK and paxillin as well as cell motility induced by the cytokine. CONCLUSIONS: Collectively, these data demonstrate for the first time that IL-6 stimulates VSMC motility which correlated with induction of actin cytoskeletal reorganization and tyrosine phosphorylation of FAK and paxillin. PMID- 12850473 TI - Trypsinogen-2 and trypsinogen activation peptide (TAP) in urine of patients with acute pancreatitis. AB - BACKGROUND AND AIMS: There is an obvious clinical need for a simple test that can identify patients at risk of developing severe acute pancreatitis. In this work we compared urinary trypsinogen-2 with urinary trypsinogen activation peptide (TAP) and serum C-reactive protein (CRP) for early differentiation between mild and severe acute pancreatitis. PATIENTS AND METHODS: The study population consisted of 127 consecutive patients with acute pancreatitis of whom 29 had severe disease. Urinary trypsinogen-2 was measured by a quantitative immunofluorometric assay and TAP by a competitive immunoassay. Serum CRP was determined by immunoturbidimetry. RESULTS: The sensitivity and specificity to identify severe acute pancreatitis on admission was 72% and 81% for urinary trypsinogen-2, 64% and 82% for urinary TAP, and 29% and 93% for serum CRP, respectively. At 24 h after admission, the values were 82% and 78% for urinary trypsinogen-2, 52% and 92% for urinary TAP, and 84% and 72% for serum CRP, respectively. Receiver-operating characteristics curve analysis showed that the area under the curve was larger for urinary trypsinogen-2 than for urinary TAP and serum CRP on admission and 24 h after admission. On admission the positive likelihood ration for urinary trypsiongen-2 was 3.7, for urinary TAP 3.6, and 4.3 for serum CRP, respectively. The corresponding negative likelihood ratios were 0.34, 0.43, and 0.76, respectively. CONCLUSION: Urinary trypsinogen-2 was superior to serum CRP and as god as or even better than urinary TAP and in the early prediction of disease severity in acute pancreatitis. These results suggest that it could be a valuable adjunct in the early assessment of the severity of acute pancreatitis. PMID- 12850474 TI - Differential responses by pancreatic carcinoma cell lines to prolonged exposure to Erbitux (IMC-C225) anti-EGFR antibody. AB - BACKGROUND: Pancreatic cancer remains a devastating disease, with 95% of all patients diagnosed with the disease dying within 2 years. The combined therapy using Erbitux, gemcitabine, and radiation caused complete tumor regression using a nude mouse model inoculated with pancreatic MiaPaCa-2 cells but only a delay in tumor growth with BxPC-3. We investigated the effect of prolonged Erbitux treatment to the sensitivity to gemcitabine and/or radiation and the epidermal growth factor receptor (EGFR) signal transduction pathway. METHODS: MiaPaCa-2 and BxPC-3 cells were cultured with or without Erbitux for 6 weeks. Cells were then treated with gemcitabine and/or radiation, harvested 48 h after treatment, and counted. Differences in EGFR expression after exposure to Erbitux were analyzed by FACS. Internalization rates of EGFR induced by Erbitux on these cell lines were determined using 125I-EGF binding assay after removal of Erbitux by acidic wash. Cell lysates were harvested after cells were stimulated with EGF, FGF, or IGF-1 respectively, and EGFR was immunoprecipitated using Erbitux. Samples were separated using SDS-PAGE and transferred to PVDF membrane. The membranes were probed with antibody against human growth factor receptor binding protein (Grb2) to detect the association of this Ras-MAPK upstream adaptor protein to EGFR. Cell lysates were also separated with SDS-PAGE and probed with rabbit anti-human PARP after samples were transferred to PVDF membrane. Expression of BAX and Bcl-(XL) were probed in the cells treated with or without Erbitux. RESULTS: Proliferation assays indicated that prolonged exposure to Erbitux increased the sensitivities of MiaPaCa-2 to gemcitabine and radiation therapy (41 +/- 16% vs 52 +/- 9% for gemcitation, 28 +/- 9 vs 39 +/- 9% for combination; P = 0.015) but not for BxPC 3. FACS analysis showed that the expressed EGFR level decreased by about 42% on MiaPaCa 2 whereas no loss was seen on BxPC-3. Expression of BAX was upregulated on MiaPaCa-2. Poly (ADP-ribose) polymerase cleavage indicated the killing was mediated by apoptosis. Immunoblots showed that Grb2 was co-immunoprecipitated with EGFR after EGF stimulation. Incubation with Erbitux blocked Grb2 binding in MiaPaCa-2 but not BxPC 3. FGF transactivated EGFR down stream Ras-MAPK in the presence or absence of Erbitux. Internalization of EGFR induced by Erbitux did not differ between MiaPaCa-2 and BxPC-3. CONCLUSIONS: 1) Association of Grb2 to EGFR in BxPC-3 induced by EGF in the presence of Erbitux indicates an alternate pathway of Ras-MAPK activation, which may be related with the tumor resistance to treatment; 2) transactivation of EGFR downstream Ras-MAPK pathway by FGF contributes the resistance to treatment; and 3) downregulation of EGFR may increase the response to therapy. PMID- 12850475 TI - Altered ATP-dependent mitochondrial Ca2+ uptake in cold ischemia is attenuated by ruthenium red. AB - BACKGROUND: Graft dysfunction as a result of preservation injury remains a major clinical problem in liver transplantation. This is related in part to accumulation of mitochondrial calcium (Ca(2+)), which has been linked to activation of proapoptotic factors. We hypothesized that cold ischemia increases mitochondrial Ca(2+) uptake in a concentration dependent fashion and that ruthenium red (RR) will attenuate these changes by inhibiting the mitochondrial Ca(2+) uniporter. METHODS: Rat livers perfused with cold University of Wisconsin (UW) solution (4 degrees C) with or without RR (10 microM) via the portal vein (n = 3 per group) were processed immediately (no ischemia) or after 24 h cold storage (24 h cold ischemia). Mitochondria were separated by differential centrifugation, and adenosine triphosphate (ATP)-dependent (45)Ca(2+) uptake was determined in the presence of ATP (5 mM), adenosine diphosphate (ADP), or adenosine 5'-beta,gamma-imidotriphosphate (AMP-PNP); variable concentrations of extramitochondrial (45)Ca(2+) were used. All measurements were performed in triplicate. Student's t test with P < 0.05 was taken as significant. RESULTS: Our data demonstrate the following: 1) ATP-dependent (45)Ca(2+) uptake in mitochondria separated from livers following 24 h of cold ischemia in UW alone was higher than in mitochondria isolated from non-ischemic livers; the increased uptake was dependent on the concentration of (45)Ca(2+) in the incubation buffer. 2) There was no difference in ATP-dependent (45)Ca(2+) uptake between nonischemic mitochondria and those separated from livers stored in UW-RR for 24 h. 3) (45)Ca(2+) uptake in mitochondria from livers subjected to 24 h of cold ischemia in UW-RR was significantly lower compared to those from livers stored in UW alone when (45)Ca(2+) concentrations were greater than 1 microM. CONCLUSION: 1) Cold ischemia affects mitochondrial Ca(2+) handling, especially when it is challenged by high extramitochondrial Ca(2+) concentrations. 2) The addition of RR in preservation solution attenuates the effects of cold ischemia on mitochondrial Ca(2+) handling. 3) Inhibition of mitochondrial Ca(2+) uniporter with RR protects mitochondria from Ca(2+) overload at high Ca(2+) concentrations. These findings may offer a potentially effective strategy for prevention of ischemia-reperfusion injury in liver transplantation. PMID- 12850476 TI - Localization of HTLV-I tax proviral DNA in mononuclear cells. AB - The tax sequence of HTLV-I is demonstrable in the skin and blood mononuclear cells of patients with mycosis fungoides, as well as in the mononuclear leukocytes of some healthy blood donors, but was not demonstrable when PCR/Southern analyses were carried out on preparations of high-molecular-weight genomic DNA. Therefore, it was postulated that tax DNA may not be integrated. To investigate this possibility fluorescence in situ hybridization was carried out on cells arrested in metaphase, using a probe containing the HTLV-I tax proviral DNA full-length open reading frame coding sequence. While metaphases prepared from C91PL cells, a cell line infected with HTLV-I, showed an abundance of chromosome-associated as well as extra-chromosomal signals, metaphases prepared with blood mononuclear cells from healthy tax sequence positive donors did not reveal any tax DNA associated with chromosomes. Such signals were readily detected extra-chromosomally. Although it has been demonstrated that transactivation of genes by gene products encoded by extra-chromosomal DNA may have nosocomial implications, whether transactivation by p40 tax generated from extra-chromosomal tax sequences is responsible for the development of neoplasia remains to be investigated. PMID- 12850477 TI - Antibody-targeted chemotherapy of acute myeloid leukemia using gemtuzumab ozogamicin (Mylotarg). AB - Acute myeloid leukemia (AML) is the most common type of acute leukemia in adults and accounts for 20% of pediatric leukemia. Although conventional chemotherapy induces clinical remissions in most patients with AML, recurrent leukemia represents the major obstacle to cure. Conventional chemotherapy reinduction is associated with limited efficacy and substantial toxicity. Chemotherapy specifically targeted to leukemic cells by monoclonal antibodies might enable patients to achieve remissions more safely than conventional approaches. After evaluating a series of phase II studies, the U.S. Food and Drug Administration approved Mylotarg (gemtuzumab ozogamicin) for the treatment of patients with CD33 positive AML in first relapse who are 60 years of age or older and who are not considered candidates for other types of cytotoxic chemotherapy. Among 277 adult patients with CD33-positive AML in first relapse, 26% experienced an overall response after Mylotarg monotherapy. Despite the fact that myelosuppression, hyperbilirubinemia, and elevated hepatic transaminases were commonly observed, the incidences of severe infections and mucositis were relatively low in comparison with conventional chemotherapeutic treatment. Preliminary reports in pediatric patients also report Mylotarg to be reasonably well tolerated. Recently, data from study regimens combining Mylotarg and conventional chemotherapy suggest an unusually high remission induction rate in de novo AML patients. Information assembled from prospective, ongoing studies in the United States and the United Kingdom should help us use this novel immunoconjugate in a safe and effective manner. PMID- 12850478 TI - Involvement of Akt kinase in the action of STI571 on chronic myelogenous leukemia cells. AB - To elucidate the role of mitogen-activated protein kinases (MAPKs) and Akt kinase in leukemogenesis caused by the breakpoint cluster region (BCR)-Abelson (ABL) tyrosine kinase oncoprotein, we examined the activities of MAPKs and Akt kinase and their roles in the action of STI571, a specific inhibitor of BCR-ABL tyrosine kinase, in chronic myelogenous leukemia (CML) cells. We found that extracellular signal-regulated kinase (ERK) 1/2 and Akt kinase are constitutively active in the chronic phase of CML, blast crisis of CML, and the CML-derived K562 cell line. Both interferon-alpha and STI571 suppressed ERK1/2 activity in K562 cells. In contrast, Akt kinase activity was inhibited only by STI571. K562 cell proliferation was markedly suppressed by LY294002, a specific inhibitor of PI3K/Akt kinase, and STI571 but not by PD98059, a specific inhibitor of MEK1/2. In addition, caspase-3 was activated by treatment of cells with STI571 and LY294002 but not with PD98059. These data indicate that Akt kinase may play a role in the proliferation of CML leukemia cells and the action of STI571. Primary leukemia cells from patients with CML blast crisis did not show inhibition of ERK1/2 or Akt kinase activity and were resistant to caspase-3-associated apoptosis after treatment with STI571. These findings suggest that STI571 does not effectively block signaling molecules downstream of the BCR-ABL tyrosine kinase in some cases of CML blast crisis. PMID- 12850479 TI - Identifying and tracking neural stem cells. AB - Hematopoietic stem cells, unlike neural stem cells, can be readily identified and isolated from developing and adult cell populations using positive and negative selection criteria. Isolating stem cells and progenitor cells from neural tissue has been more difficult because of difficulties in separating cells in solid tissue, the limited numbers of stem cells that persist in the adult, and the paucity of rigorously characterized markers. Nevertheless, strategies that have worked successfully in hematopoietic stem cell isolation can be adapted to isolate multiple classes of stem and progenitor cells from neural tissue. Neural stem cells also share cellular and molecular properties with other stem cell populations that may serve as surrogate identifiers of multipotentiality. Such potential markers are described. Unlike hematopoietic stem cells, tracking neural cells after transplantation is both necessary and more difficult. It will therefore be necessary to develop invasive and non-invasive strategies to follow transplanted cells and develop useful quantifiable readouts. Some potential strategies are described and current results are discussed. PMID- 12850480 TI - The use of lentiviral vectors in gene therapy of leukemia: combinatorial gene delivery of immunomodulators into leukemia cells by state-of-the-art vectors. AB - Our goal is to develop cell vaccines against leukemia cells, genetically modified to express molecules with potent immune-stimulatory capacities. Pre-clinical evaluation of this approach in murine models has demonstrated efficient anti leukemic responses with the expression of immunomodulators, in particular GM-CSF and CD80, in irradiated cell vaccines. We have previously shown efficient insertion of GM-CSF and CD80 genes into primary human leukemia cells with the use of second and third generation self-inactivating (SIN) lentiviral vectors (Blood 96 (2000), 1317; Leukemia 16 (2002), 1645). The advantages of lentiviral vectors for development of autologous leukemia cell vaccines include: (1) efficient and consistent gene delivery; (2) high levels of transgene expression; (3) persistent expression of the transduced gene; (4) no viral proteins, as only the transduced gene is expressed; (5) no undesirable cytotoxic effects, and; (6) simplicity of use [leukemia cells are exposed to vector(s) only once]. In this work, we evaluated the insertion of the central polypurine tract and the central termination sequence into a SIN lentiviral vector encoding for GM-CSF and CD80, which significantly enhanced the transduction efficiency of primary leukemia cells and provided higher levels of GM-CSF and CD80 co-expression. We also demonstrate a methodology to deliver simultaneously a combination of immunomodulatory molecules (GM-CSF, CD80, IL-4, and CD40L) to activate different pathways of immune stimulation. Therefore, lentiviral vectors offer a simple, versatile, and reliable approach for engineering leukemic cells for use as cell vaccines. PMID- 12850481 TI - Analysis of the A(TA)(n)TAA configuration in the promoter region of the UGT1 A1 gene in Greek patients with thalassemia intermedia and sickle cell disease. AB - Gilbert's syndrome is characterized by mild unconjugated hyperbilirubinemia. The molecular basis of this syndrome usually concerns an additional dinucleotide insertion (TA) in the A(TA)(n)TAA configuration residing in the promoter region of the UGT1 A1 gene. This configuration may vary in length; the "n" represents the different number of TA repeats. The homozygosity A(TA)(7)TAA/A(TA)(7)TAA is involved in Gilbert's syndrome. In many cases of patients with thalassemia intermedia and sickle cell disease considerable variation in bilirubin levels is observed. In this study we investigated the contribution of the A(TA)(7)TAA/A(TA)(7)TAA genotype in the variable unconjugated serum bilirubin levels in 31 Greek patients with thalassemia intermedia and 27 Greek compound heterozygotes for beta thalassemia and sickle cell anemia. Analysis of the A(TA)(n)TAA configuration in the promoter region of the latter patients showed that those who were carrying the homozygosity A(TA)(7)TAA/A(TA)(7)TAA had higher levels of unconjugated bilirubin. These findings suggest that the coexistence of Gilbert's syndrome in patients with thalassemia intermedia and sickle cell disease may be the cause of the elevated values of unconjugated bilirubin, reducing the possibility of excessive hemolysis in these patients. PMID- 12850482 TI - Osmotic swelling and hole formation in membranes of thalassemic and spherocytic erythrocytes. AB - Osmotic swelling and kinetics of the pore formation in the membranes of spherocytic, thalassemic, and normal erythrocytes were studied by measuring the time-dependent capacitance and conductance at a frequency of 0.2 MHz. No significant difference between the swelling rate of control and spherocytic cells was observed, whereas slower kinetics of swelling were found for thalassemic cells. Time records of the conductance indicate that the probability of the pore formation in the stretched membrances varies in the following order: thalassemia < control < spherocytosis. Based on these findings it was concluded that the erythrocyte swelling is controlled by the initial cell shape, volume, intracellular hemoglobin concentration, and elastic membrane properties, whereas the kinetics of the pore formation depend solely on the resistivity of the stretched membrane of the swollen RBC to the osmotic shock. Therefore, it was assumed that investigations of the pore formation may be used not only for examinations of spherocytic and thalassemic cells, but also for normocytic, normochromic, biconcave-shaped RBCs with altered membrane elasticity. PMID- 12850483 TI - SarCNU mediates selection of P140K methylguanine-DNA-methyltransferase transduced human CD34(+) cells in vitro. AB - The therapeutic combination O(6)-benzylguanine (BG) and 1,3-bis(2-chloroethyl)-1 nitrosourea (BCNU) can be used to select for P140K methylguanine-DNA methyltransferase (P140K MGMT) transduced hematopoietic progenitors both in vitro and in vivo. SarCNU is a new alkylating agent which has advantages over BCNU. We tested the ability of P140K MGMT transduced human CD34(+) cells to resist SarCNU treatment and be selected for in vitro. BG/SarCNU exposure led to an increase in the proportion of transduced cells from 13 to 27%. The IC(90) was increased sixfold by P140K MGMT transduction. These results suggest that SarCNU may be a suitable agent for in vivo selection strategies. PMID- 12850484 TI - Regulatory issues in human gene therapy. AB - Protection of human subjects is the ethical and legal responsibility of investigators conducting clinical trials. Public concerns regarding recombinant DNA technology led to additional levels of oversight which are unique to human gene therapy trials. The deaths of a normal volunteer and a gene therapy subject in the late 1990s led to an intensification of oversight with new initiatives that impact gene therapy as well as other clinical investigators. This paper will review the current oversight agencies and identify areas of evolving regulations that pose particular challenges to gene therapy investigators. PMID- 12850486 TI - Immune monitoring of human gene therapy trials: potential application to leukemia and lymphoma. AB - Therapeutic genetically modified vaccines administered to patients with cancer are expected to induce and sustain tumor-specific immune responses. To measure these responses, monitoring strategies including a range of antibody-based and cellular assays have been developed. Single-cell cellular assays capable of detecting fewer than 1/10,000 of antigen-specific T lymphocytes in the tested population are currently in demand. ELISPOT assay, cytokine flow cytometry, and tetramer binding were recently compared by us in the context of monitoring of a dendritic cell-based multi-peptide vaccination trial in patients with metastatic melanoma. These single-cell assays were not found to be concordant in measurements of the frequency of peptide-specific T cells or in detecting pre- to post-vaccine changes in the frequency of these T cells. Implications of the results for monitoring of genetically modified vaccines are discussed, and the plan for a rational approach to monitoring and for selection of monitoring assays is proposed. Monitoring of human gene therapy or vaccination trials in patients with advanced cancers, who are often immunocompromised and thus poorly responsive, is a challenge. This challenge is best met by utilizing experienced reference laboratories for immune monitoring of clinical trials. PMID- 12850485 TI - Association between HFE mutations and acute myocardial infarction: a study in patients from Northern and Southern Italy. AB - There is interest in the role of iron in age-related diseases such as atherosclerosis. Tissue iron deposition could be harmful, because Fe(2+) can react with H(2)O(2) to form OH(-) radicals and Fe(2+) can react with O(2) to form reactive oxygen species. Free radicals react with cell membranes and cell organelles and could lead to the development of atherosclerosis by initiating lipid peroxidation. Hereditary hemochromatosis provides an opportunity for studying the effects of iron on cardiovascular disease. Some studies have shown that individuals who carried HFE mutations may be at greater risk of developing coronary heart disease than those without the mutations. In contrast, a large number of studies have reported no association between HFE mutations and coronary heart disease. These studies have possible confounding factors, such as the homogeneity of the population in term of geographical origin among others. We studied the relation between HFE mutations and acute myocardial infarction in two case-control studies involving two sets of subjects representing different age groups from different geographic regions in Italy. The first one was composed of 172 older patients (139 males and 33 females; mean age 67) and 207 healthy controls (91 males and 116 females; mean age 46) from Emilia-Romagna. The second one was composed of younger 77 patients (75 males and 2 females; mean age 41) and 172 healthy controls (75 males and 97 females, mean age: 38) from Sicily. All patients were genotyped for ApoE alleles, since the ApoE- epsilon 4 allele is considered a risk factor for cardiovascular diseases and can interfere with other genetic and environmental factors by modifying susceptibility to this disease. DNA typing for C282Y and H63D HFE alleles was performed also. There were no significant differences in frequencies of the different HFE alleles between acute myocardial infarction patients and controls in cohorts of both old and young patients. Also taking into account the presence or absence of the ApoE- epsilon 4 allele, no significant differences in H63D allele frequencies were observed. Thus, our study, performed in two samples of genetically homogeneous patients and controls, does not support the suggestion that HFE mutations may be associated with acute myocardial infarction in susceptible individuals. PMID- 12850487 TI - The 1226G (N370S) Gaucher mutation among patients with Legg-Calve-Perthes disease. AB - Legg-Calve-Perthes disease (LCPD) is an avascular necrosis of the femoral head with an annual incidence of 5-15/100,000. The estimated incidence of Gaucher disease, a lysosomal recessive storage disease, is 1:850, with a carrier rate of 1:17.5 for the 1226G (N370S) mutation among Ashkenazi Jews in whom there is a predilection. Since clinical and radiological findings of avascular hip necrosis due to either Gaucher disease or LCPD may be indistinguishable, misdiagnosis may occur. The purpose of this study was to evaluate the incidence of 1226G Gaucher mutation in a cohort of radiologically confirmed LCPD patients (diagnosed 1986 2000) in Israel. Enzyme assay was performed for confirmation of affected versus carrier status in patients with the 1226G mutation. In all, 78 LCPD patients, 86% males, 51% with severe bone disease, were studied. Family history was negative for Gaucher disease. Ethnic origin was 39% Ashkenazi Jewish, 6% Arab, and 55% other ethnicities. One Ashkenazi Jewish LCPD patient was homozygous for the 1226G mutation, and 4 LCPD patients were carriers: 3 Ashkenazi Jewish and 1 Arab patient. The frequency of the 1226G mutation among the LCPD patients was increased relative to historical Ashkenazi Jewish Israeli controls (P = 0.01). Since Gaucher disease may be misdiagnosed as LCPD, glucocerebrosidase enzyme testing is recommended among Ashkenazi Jewish children diagnosed with LCPD. PMID- 12850488 TI - Memory to the hapten in non-immediate cutaneous allergic reactions to betalactams resides in a lymphocyte subpopulation expressing both CD45RO and CLA markers. AB - The cutaneous lymphocyte-associated antigen (CLA) is a homing receptor expressed in a subpopulation of memory T lymphocytes that migrates to the skin and participates in different inflammatory processes. The aim of the study was to compare the T cell response to betalactams in both CLA+ and CLA- memory T cell subsets from subjects with non-immediate allergic reactions to these drugs. Three patients with a non-immediate reaction to penicillins were studied during their acute episodes. Peripheral blood mononuclear cells were isolated by Ficoll density gradient and were used for flow cytometry and lymphocyte transformation test assays. CD3+ cells were purified via high affinity negative selection columns. CD45RA+ and CD45RO+ subpopulations were obtained by magnetic sorting and the memory subpopulation was subdivided into CLA+ and CLA- fractions. These were cultured in triplicate together with feeder cells and different concentrations of amoxicillin and benzylpenicillin. In all cases, the proliferative responses to the drugs were confined to the CD45RO+CLA+ subpopulation. The CD45RO+CLA- subset showed no proliferative response to either drug at any concentration. We have shown that the in vitro memory to penicillins in non-immediate cutaneous allergic reactions to these drugs resides in the CD45RO+CD3+ subset expressing CLA, which enables these T cells to migrate to the skin. These findings may have relevance to understanding the involvement of T cells in allergic reactions to penicillins. PMID- 12850489 TI - Enhancing dendritic cell-based vaccines by gene modification. PMID- 12850490 TI - Late effects among survivors of leukemia during childhood and adolescence. AB - Acute lymphoblastic leukemia (ALL) is the most common childhood malignancy, accounting for one-third of all cancers occurring in childhood and adolescents. Over the last 3 decades there has been a marked improvement in survival, with 5 year survival rates approaching 80%. With this improvement in survival, increasing attention is now being focused on issues relating to long-term morbidity and mortality associated with the treatments responsible for that increased survival. Because of the young age of these cancer survivors, and thus the potential longevity, the delayed consequences of therapy may have a significant impact on their lives. Long-term sequelae of treatment, such as impaired intellectual and psychomotor functioning, neuroendocrine abnormalities, impaired reproductive capacity, cardiotoxicity, and second malignant neoplasms, are now being reported with increasing frequency in this growing cohort of survivors and knowledge of the late -effects associated with cancer in children and adolescents continues to increase through ongoing research efforts. However, much of the available information relates to outcomes within the first decade following treatment, although information about the longer term outcomes that may occur later in adulthood is emerging as a result of well-conducted, large cohort studies. Through a multi-disciplinary approach to the diagnosis, treatment, and long-term follow-up of pediatric leukemia patients, we can achieve the goal of cure while minimizing the occurrence of long-term adverse outcomes. This review summarizes some of the well-described long-term consequences of therapy among children and adolescents treated for ALL. PMID- 12850491 TI - Diamond blackfan anemia stem cells fail to repopulate erythropoiesis in NOD/SCID mice. AB - Diamond Blackfan Anemia (DBA) is a congenital disorder characterized by decreased red blood cell production and developmental abnormalities. We herein show that DBA progenitors produced lower numbers of phenotypically normal erythroid colonies in vitro, whereas nonerythroid colonies were normally abundant and developed. To determine whether DBA stem cells are capable of producing early erythroid, monocyto-granulocytic, and lymphoid progenitors in vivo we used a mouse xenotransplantation model. We demonstrate that DBA stem cells poorly repopulated erythroid progeny in NOD/SCID mice, whereas the monocyto-granulocytic and lymphoid progenies were repopulated normally. Therefore, we conclude that disordered DBA erythropoiesis may be a result of defective erythroid-lineage commitment and maintenance of early erythroid progenitors. PMID- 12850492 TI - The global distribution of length polymorphisms of the promoters of the glucuronosyltransferase 1 gene (UGT1A1): hematologic and evolutionary implications. AB - The promoter region of the UDP glucuronosyltransferase 1 gene (UGT1A1) contains a run of thymine-adenine (TA) repeats, usually six (TA)(6). As well as its relationship to Gilbert's syndrome, homozygosity for the extended sequence, (TA)(7) (TA)(7), has been found to be an important risk factor for hyperbilirubinemia and gallstones in patients with hemoglobin E-beta-thalassemia and other intermediate forms of beta thalassemia. To assess the importance of this polymorphism in these common disorders a wide-scale population study of the relative frequency of the size alleles of the UGT1A1 promoter has been carried out. Homozygosity for the (TA)(7) allele occurs in 10-25% of the populations of Africa and the Indian subcontinent, with a variable frequency in Europe. It occurs at a much lower frequency in Southeast Asia, Melanesia, and the Pacific Islands, ranging from 0 to 5%. African populations show a much greater diversity of length alleles than other populations. These findings define those populations with a high frequency of hemoglobin E-beta-thalassemia and related disorders that are at increased risk for hyperbilirubinemia and gall bladder disease and provide evolutionary insights into how these polymorphisms have arisen and are so unequally distributed among human populations. PMID- 12850493 TI - Common variable immunodeficiency and IgG subclass deficiency in central Alabama hemochromatosis probands homozygous for HFE C282Y. AB - Eight hemochromatosis probands with HFE C282Y homozygosity had frequent, severe, or unusual infections and common variable immunodeficiency (CVID) or immunoglobulin (Ig) G subclass deficiency (IgGSD). Thus, we performed serum Ig isotyping and other characterization of 43 additional unselected probands, 5 human leukocyte antigen (HLA)-identical siblings, and 240 consecutive CVID or IgGSD index patients. C282Y allele frequencies were estimated in 58 CVID or IgGSD index patients without hemochromatosis phenotypes and in 341 controls. HLA-A and B haplotypes and frequencies were determined in all 51 probands, 186 CVID or IgGSD index patients without hemochromatosis phenotypes, and 751 controls. Thirteen unselected probands (30%) had CVID or IgGSD. Among all 21 hemochromatosis probands with CVID (n = 4) or IgGSD (n = 17), Ig subclass deficiency patterns were IgG(1) (n = 5), IgG(1) and IgG(3) (n = 6), IgG(3) (n = 9), and IgG(1), IgG(3), and IgG(4) (n = 1). IgG(2) or IgA deficiency was not detected; one proband had IgM deficiency. Mean values of total IgG, IgG(1), and IgG(3) were significantly lower in probands with CVID or IgGSD. Mean values of age, transferrin saturation, and ferritin at diagnosis and phlebotomy units required to induce iron depletion were similar in probands with or without CVID or IgGSD; phlebotomy had no apparent effect on IgG levels. C282Y frequencies were similar in CVID or IgGSD index cases without hemochromatosis phenotypes and in controls. There was concordance of Ig and hemochromatosis phenotypes in probands and respective HLA-identical siblings. Eight of 240 CVID or IgGSD index patients had hemochromatosis phenotypes and C282Y homozygosity (3 vs 0.7% and 0.2% controls; P < 0.0001, respectively). The frequency of A*03-B*07 was greater in CVID and IgGSD index cases without hemochromatosis phenotypes than in controls (0.0968 vs 0.0546, respectively; P = 0.0032). HLA-A*03-B*07 was the predominant haplotype in probands grouped by presence or absence of CVID or IgGSD. Some probands in each group were A*03-B*07 homozygotes; group A*03-B*07 frequencies were similar. We conclude that serum IgG abnormalities characteristic of CVID or IgGSD are common in hemochromatosis probands, and that the prevalence of hemochromatosis is increased in CVID and IgGSD index cases. These observations could be explained by the increased frequencies of HLA-A*03-B*07 in C282Y homozygotes and in CVID and IgGSD, and by the common occurrence of putative CVID or IgGSD allele(s) on haplotypes bearing C282Y. PMID- 12850494 TI - Glucose-6-phosphate dehydrogenase (G-6-PD) mutations in Mexico: four new G-6-PD variants. AB - Screening for mutations at the G-6-PD gene by PCR-SSCP combined with restriction enzyme analysis and DNA sequencing was performed in nine G-6-PD deficient individuals with negative results for the presence of the most frequent G-6-PD mutations previously observed in Mexican population. The variants G-6-PD Valladolid(406T), G-6-PD Durham(713G), and G-6-PD Viangchan(871A) and four new G 6-PD mutant alleles were identified. The new mutations are located at cDNA nt 376 A --> T (126 Asn --> Tyr), nt 770 G --> T (257 Arg --> Leu), nt 1094 G --> A (365 Arg --> His), and nt 1285 A --> G (429 Lys --> Glu) and they were named G-6-PD San Luis Potosi, G-6-PD Zacatecas, G-6-PD Veracruz, and G-6-PD Yucatan, respectively. To date, a total of 18 different G-6-PD variants have been observed in Mexico and several of them are common in Africa, South Europe, and Southeast Asia. PMID- 12850495 TI - Autologous immunotherapy for human leukemias. AB - Immunotherapy for human leukemias has the potential to contribute to the long term control or cure of these diseases. Our work demonstrates that cells from the majority of adult acute myelogenous leukemia cases can be induced to differentiate into dendritic cells that are effective at antigen presentation. Both interleukin-4 and CD40 ligand are important for optimal dendritic cell differentiation and maturation. Granulocyte-monocyte colony-stimulating factor, interleukin-4, and CD40 ligand in combination are capable of yielding dendritic cells from at least some cases of acute lymphocytic leukemia. Efforts to clone autologous, cytotoxic effector cells will permit the identification of target antigens in the future. With information concerning potential antigens, protocols inducing antileukemic immunity should be possible. Prior to that time, with the availability of suitable reagents for clinical scale differentiation of leukemia derived dendritic cells, such cells might prove potent as vaccines for the therapy of acute leukemias. PMID- 12850497 TI - Discriminative stimulus effects of m-chlorophenylpiperazine as a model of the role of serotonin receptors in anxiety. AB - Serotonin is known to play a role in anxiety. The roles of serotonin reuptake and 5-HT1A receptors have been well characterized, but the contribution of other serotonin receptor subtypes is not as clear. 1-(3-Chlorophenyl)-piperazine (mCPP), which binds non-selectively to a wide range of serotonin receptors, has often been used to produce anxiety in humans and in animal models. Because functional assays indicate that mCPP is significantly more potent at 5-HT2C receptors, it may serve as a tool to investigate the contribution of 5-HT2C receptors to anxiety. This paper reviews the results of behavioral tests using mCPP, including the drug discrimination assay, to model anxiety. Although the discriminative stimulus effects of mCPP do not seem to be a useful screen for general anxiolytics, they do seem to be useful for characterization of the contribution of 5-HT1B and 5-HT2C receptors to the mediation of anxiety-like behaviors. PMID- 12850498 TI - Modulation of IR/PTP1B interaction and downstream signaling in insulin sensitive tissues of MSG-rats. AB - PTP1B has been shown to be a negative regulator of the insulin signal transduction in insulin resistant states. Herein we investigated IR/PTP1B interaction and downstream signaling in insulin sensitive tissues of 10 and 28 week-old MSG-insulin resistant rats which represent different stages of insulin resistance. Our results demonstrated that the increase in PTP1B expression and/or association with IR in MSG animals may contribute to the impaired insulin signaling mainly in liver and muscle. Although, adipose tissue of 10-week-old MSG rats showed higher PTP1B expression and IR/PTP1B interaction, they were not sufficient to impair all insulin signaling since IRS-2 phosphorylation and association with PI3-kinase and Akt serine phosphorylation were increased, which may contribute for the increased adiposity of these animals. In 28-week-old-MSG rats there was an increase in IR/PTP1B interaction and reduced insulin signaling in liver, muscle and adipocytes, and a more pronounced insulin resistance. PMID- 12850499 TI - Stimulatory effect of oral administration of green tea and caffeine on locomotor activity in SKH-1 mice. AB - Administration of green tea or caffeine was shown previously to inhibit ultraviolet B light-induced carcinogenesis in SKH-1 mice, and this effect was associated with a reduction in dermal fat. In the present study, oral administration of 0.6% green tea (6 mg tea solids/ml) or 0.04% caffeine (0.4 mg/ml; equivalent to the amount of caffeine in 0.6% green tea) as the sole source of drinking fluid to SKH-1 mice for 15 weeks increased total 24 hr locomotor activity by 47 and 24%, respectively (p<0.0001). Oral administration of 0.6% decaffeinated green tea (6 mg tea solids/ml) for 15 weeks increased locomotor activity by 9% (p<0.05). The small increase in locomotor activity observed in mice treated with decaffeinated green tea may have resulted from the small amounts of caffeine still remaining in decaffeinated green tea solutions (0.047 mg/ml). The stimulatory effects of orally administered green tea and caffeine on locomotor activity were paralleled by a 38 and 23% increase, respectively, in the dermal muscle layer thickness. In addition, treatment of the mice with 0.6% green tea or 0.04% caffeine for 15 weeks decreased the weight of the parametrial fat pad by 29 and 43%, respectively, and the thickness of the dermal fat layer was decreased by 51 and 47%, respectively. These results indicate that oral administration of green tea or caffeine to SKH-1 mice increases locomotor activity and muscle mass and decreases fat stores. The stimulatory effect of green tea and caffeine administration on locomotor activity described here may contribute to the effects of green tea and caffeine to decrease fat stores and to inhibit carcinogenesis induced by UVB in SKH-1 mice. PMID- 12850500 TI - Hypolipidemic action of dietary resveratrol, a phytoalexin in grapes and red wine, in hepatoma-bearing rats. AB - Resveratrol is an antioxidant present in grapes and their related products. We investigated whether dietary resveratrol could inhibit the proliferation and metastasis of tumors and hyperlipidemia in Donryu rats subcutaneously implanted with an ascites hepatoma cell line of AH109A. By feeding 10 or 50 ppm resveratrol in the diet to hepatoma-bearing rats for 20 days, solid tumor growth and metastasis tended to be suppressed dose-dependently. Resveratrol (50 ppm) significantly suppressed the serum lipid peroxide level, indicating its antioxidative properties or those of its metabolite(s) in vivo. Resveratrol dose dependently suppressed both the serum triglyceride and very-low-density lipoprotein + low-density lipoprotein (VLDL + LDL)-cholesterol levels. The hypocholesterolemic action of resveratrol is attributed, at least in part, to an increased excretion of neutral sterols and bile acids into feces. These results suggest that dietary resveratrol is hypolipidemic with a tendency for anti-tumor growth and anti-metastasis effects in hepatoma-bearing rats. PMID- 12850501 TI - Effects of 13-alkyl-substituted berberine alkaloids on the expression of COX-II, TNF-alpha, iNOS, and IL-12 production in LPS-stimulated macrophages. AB - Berberine, a major alkaloidal component of Coptidis Rhizoma, has antibacterial activity, anti-inflammatory effect, antitumor and antimotility actions. We suggested that one of possible mechanisms of anti-bacterial activity of berberine may be based on the production of interleukin (IL)-12. Recently 13-alkyl substituted berberines were shown to be better activity than berberine against certain bacteria species and human cancer cell lines. In the present study, therefore, the effects of 13-methylberberine (13-MB) and 13-ethylberberine (13 EB) on the production of IL-12 and expression of iNOS, TNF-alpha and COX-II were investigated using macrophages in culture. In LPS-stimulated RAW 264.7 cells, these alkaloids decreased the nitrites, concentration-dependently. The concentration of 50% inhibition of NO production (IC50) by 13-MB and 13-EB was 11.64 and 9.32 microM, respectively. The suppressed expression of iNOS protein was responsible for the reduction of NO production. Neither the expression of mRNA of iNOS, COX-II and TNF- alpha nor protein of COX-II and TNF-alpha was affected by both 13-MB and 13-EB, but production of PGE2 in LPS-stimulated RAW 264.7 cells was significantly reduced. Another striking finding of the present study is that 13-MB and 13-EB increased production of IL-12 in LPS-treated splenic macrophages. These results indicate that posttranscriptional regulatory mechanism of iNOS gene expression by 13-MB and 13-EB is involved, and COX-II activity is inhibited by 13-MB and 13-EB, respectively. In conclusion, the present study demonstrates that 13-methyl- and 13-ethylberberine alkaloids can be useful as an immunotherapeutic compound for induction of IL-12, which is potentially applicable for tumors, infectious disease, and airway inflammation. PMID- 12850503 TI - Expression of VEGF and MMP-9 in giant cell tumor of bone and other osteolytic lesions. AB - This study aims to investigate the expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in giant cell tumor of bone (GCT) and other osteolytic lesions in bone. By using semi-quantitative RT-PCR, we showed that three major isoforms of VEGF (121, 165 and 189) were expressed in GCTs, with isoform 121 being the most abundant. The expression levels of VEGF and MMP-9 mRNA were significantly higher in advanced GCTs (stage II/III) than in stage I GCTs. We further elucidated the cellular localization of VEGF and MMP-9 gene transcripts in GCT and other osteolytic lesions using an in situ hybridization assay. The results showed that stromal tumor cells and osteoclast like giant cells of GCT, fibrous stromal cells in anuerysmal bone cysts and fibrous dysplasia, and Langerhans-type giant cells as well as histocytes in eosinophillic granuloma, were all strongly positive for VEGF and MMP-9 mRNA expression. In a prospective study, we performed VEGF and MMP-9 immuno-staining on paraffin sections of pathological tissues harvested from 48 patients (14 GCT, 10 anuerysmal bone cysts, 10 eosinophillic granuloma, 4 fibrous dysplasia, 2 simple bone cyst, 2 osteomyelitis and 6 patients with fractured femoral head as control). The results showed that the differences in VEGF and MMP-9 expression between Stage I and other advanced Stages (II, III and recurrent) were highly significant (p<0.001), with advanced stages showing a higher mean expression. The difference between recurrent and Stage II and III lesions, was also statistically significant (p=0.03 for VEGF, and p=0.01 for MMP-9 expression), with recurrent lesions showing a higher mean expression of both VEGF and MMP-9. In conclusion, VEGF and MMP-9 expression in osteolytic lesions of bone co-relates well with the extent of bone destruction and local recurrence. Their expression may therefore provide some prognostic indication of the possible aggressive behavior of the underlying pathology. PMID- 12850502 TI - Comparison of cardioprotective effects using ramipril and DanShen for the treatment of acute myocardial infarction in rats. AB - In the present study, we compared cardioprotective effects of DanShen (an extract from Salvia miltiorrhiza) and the angiotensin-converting enzyme inhibitor, ramipril, in rats. With both treatment regimens, DanShen- and ramipril similar effects were observed: (1) a higher survival rate, (2) a significant reduction of infarct size, (3) significantly lower ratios of heart weight to the body weight as well as the left and right ventricular weights to body weight. DanShen showed some unique effects in the following aspects: (1) higher activities of antioxidant defense enzymes such as superoxide dismutase (SOD), catalase (CAT), glutatione perioxidase (GSH-Px) and glutathione S-transferase (GST) in the liver of rats with acute myocardial infarction (AMI), (2) lower myocardial and hepatic TBARS values; (3) augmented VEGF mRNA expressions in the non-ischemic parts of rat hearts with AMI. These results were consistent with the findings of a slight increase in myocardial capillary density and the special distribution pattern of coronary blood vessels in DanShen-treated rats. PMID- 12850504 TI - Microdialysis and 2-[18F]fluoro-2-deoxy-D-glucose (FDG): a study on insulin action on FDG transport, uptake and metabolism in rat muscle, liver and adipose tissue. AB - A combination of microdialysis (MD) and 2-[18F ]fluoro-2-deoxy-D-glucose (FDG) was used to assess FDG uptake, phosphorylation and the glucose metabolic index (Rg') in certain tissues of fed and fasting anesthetized Sprague-Dawley rats which received an i.v. bolus injection of insulin or saline during the course of the study. The relative recovery for FDG for the MD probes was also measured as a function of flow rate and temperature. The elimination half-life (T(1/2 FDG)) of FDG from the plasma and the extracellular fluid of muscle and liver was studied with MD. The phosphorylation of FDG in muscle, liver, subcutaneous fat and mesenteric fat from homogenates of these tissues was analyzed by a radioHPLC method and the Rg' was calculated. The results show that the nutritional status does not affect the T(1/2 FDG), the total uptake of FDG 6-phosphate or the Rg' values in the studied tissues at ambient glucose. Insulin stimulation decreased T(1/2 FDG), and increased the total FDG 6-P accumulation and Rg' in the muscle of fed and fasted rats. In adipose tissues the insulin stimulation enhanced the phosphorylation but in muscle the proportion of FDG 6-P remained unchanged. Rg' in adipose tissue was higher after insulin administration in fed rats than without insulin but with fasted rats there were no differences in Rg' values with or without insulin, although the proportion of FDG 6-P did increase. The Rg' values for the livers were unaffected by any of the manipulations, but fasted rats accumulated proportionately more FDG 6-P after insulin administration than did fed rats. These results indicate that the combination of MD and FDG is a valuable and reliable tool when studying glucose metabolism in physiological and pathological models in vivo. PMID- 12850505 TI - Effects of hyperoxia and acrylonitrile on the phospholipase C isozyme protein levels in rat heart and brain. AB - We previously showed that hyperoxia exerts oxidative stress on the rat cerebral cortex, and the protein levels of phospholipase C (PLC) -beta1 and -delta1, but not PLC-gamma1, were changed. Acrylonitrile (ACN) appears to induce astrocytomas through induction of oxidative stress on the rat brain selectively. This study compared hyperoxia or ACN treatments of rats with respect to lipid peroxidation and PLC levels in the heart and cerebral cortex. Treatment of rats with ACN promoted lipid peroxidation in the heart and cerebral cortex, the percent increase above control being greater in the cortex than heart. Hyperoxia did not cause significant increases in lipid peroxidation in the cerebral cortex or heart. In the ACN-treated cerebral cortex, significant increases in the PLC-beta1 and -delta1 in the cytosol, and PLC-gamma1 in the cytosolic and particulate fractions, and lysate were observed. In the rat heart, in which PLC-beta1 could not be detected, PLC-gamma1 and -delta1 were increased and decreased in the cytosolic and particulate fractions, respectively, by hyperoxia. In addition, the expression level of PLC-gamma1 was decreased in the lysate by the treatment. In the heart treated with ACN, there was no change in the level of PLC-gamma1, while PLC-delta1 was elevated in all fractions. These findings suggested that the expression levels of PLC isozymes are altered by hyperoxia and ACN, but there are apparent differences in these altered levels between the different levels of oxidative stress, and between the organs. PMID- 12850506 TI - Impact of an enriched-cholesterol diet on enzymatic cholesterol metabolism during rabbit gestation. AB - An appropriate cholesterol homeostasis is vital for the maintenance and the optimal fetal development. The cholesterol is essential for the synthesis of progesterone and 17beta-estradiol, hormones that actively participate to sustain gestation. However, the administration of 0.2% enriched cholesterol diet (ECD) during rabbit gestation significantly increased the cholesterol blood profile (total-cholesterol, LDL, HDL, esterified-cholesterol and free-cholesterol) of dams and offspring, and induced a reduction of the offspring weight of 15% as compared to the control group. Enzymes involved in cholesterol metabolism (ACAT, HMG-CoA-reductase and cholesterol-7alpha-hydroxylase) are greatly influenced by cholesterol profile. We hypothesized that the administration of an ECD during rabbit gestation modifies the activity of those enzymes. Female rabbits (pregnant or not) were fed with a standard diet or an ECD. At term, livers (dams and offspring) and placentas were collected and ACAT, HMG-CoA-reductase and cholesterol-7alpha-hydroxylase activities were assayed. Our results demonstrate that gestation induced a reduction of ACAT activity (48.9%) in dam's liver and, an augmentation of HMG-CoA-reductase activity (142.4%) whereas it has no effect on cholesterol-7alpha-hydroxylase activity. The administration of the ECD has no additive effect on ACAT, but significantly reduced the HMG-CoA-reductase activity and cholesterol-7alpha-hydroxylase activity as compared with the pregnant control group. In placentas the ECD supplementation has an influence for HMG-CoA reductase activity, where a 43% increased in observed. Any ACAT activity was detected in placenta and the ECD has no influence on the cholesterol-7alpha hydroxylase activity. Whereas their offspring's liver present a reduction of ACAT and HMG-CoA-reductase activity. Gestation associated with ECD reduces significantly the HMG-CoA-reductase activity, decreasing the cholesterol synthesis, but placenta seems to compensate this effect by increasing its HMG-CoA reductase activity. PMID- 12850507 TI - Life and death of children: the price of life. AB - Nearly 2 million children die of simple diarrhea each year in developing countries. Those children would not have died if they were given Oral Rehydration Solution, which costs 100 yen for treating one child. In developed countries like Japan, treating one patient with intracranial aneurysm will cost over 2 million yen. The cost for treating one such patient could save 10,000 children with diarrhea even after adjusting with purchasing power parity in developing countries. Such inequity and inequality cannot be acceptable. We who are concerned about such a status quo have to work together for the betterment of children living in developing countries. In this review the author explores the root causes behind such unjust global structures. PMID- 12850508 TI - The genetics of febrile seizures and related epilepsy syndromes. AB - Febrile seizures (FS) may represent the most common seizure disorder in childhood and are known to be associated with putative genetic predispositions. Nevertheless, molecular genetic approaches toward understanding FS have been just initiated this decade. Recently, several genetic loci for FS have been mapped thereby assuring the genetic heterogeneity of FS. However, the exact molecular mechanisms of FS are yet to be elucidated. Genetic defects have been recently identified in autosomal dominant epilepsy with FS plus or generalized epilepsy with FS plus. The underlying mutations were found in genes encoding several Na+ channel subunits and the gamma2 subunit of gamma amino-butyric acid (GABA)A receptors in the brain. Furthermore, both channels are also associated with severe myoclonic epilepsy in infancy, where the seizure attacks often begin with prolonged FS and are precipitated by fever even afterwards. Na+ channels are associated with other temperature-sensitive disorders, and GABA(A) receptors are known to play an important role in the pathogenesis of FS. These lines of evidence suggest the involvement of various Na+ channels, GABA(A) receptors and additional auxiliary proteins in the pathogenesis of frequent FS and even in simple FS. This hypothesis may facilitate our understanding of the genetic background of FS. PMID- 12850509 TI - Neonatal behavioral assessment scale as a predictor of later developmental disabilities of low birth-weight and/or premature infants. AB - The purpose of this study was to evaluate the usefulness of the Neonatal Behavioral Assessment Scale (NBAS) as a tool to assess the risk of later developmental disabilities. The study subjects were 209 low birth-weight and/or premature infants admitted to the NICU at the Nagasaki University Hospital, Nagasaki, Japan. These infants were examined using the NBAS at 36-38 (NBAS36), 40 42 (NBAS40) and 44-46 weeks (NBAS44) of postmenstrual age, and their developmental outcome was measured using standardized assessments at 5 years of age. Based on the results of diagnosis at 5 years of age, subjects were classified into three groups: Normal, Mild Disability and Severe Disability groups. Multinomial logistic regression analysis showed that lower Motor cluster scores for all assessment periods and lower Orientation cluster scores in the NBAS40 and NBAS44 were significantly associated with an increased risk of both Mild and Severe Disability. Also, the Range of State cluster scores for the NBAS44 were significantly related to the risk of Mild Disability, and the Reflexes cluster scores in the NBAS40 and NBAS44 were the best predictor of Severe Disability. In outcome prediction using the estimated regression coefficients, 94-97% of the subjects in the Normal group, 50-78% in the Mild Disability group and 71-85% in the Severe Disability group were correctly classified. The NBAS could help clinicians to develop a management protocol for infants at risk for developmental disabilities as well as to identify neonates at risk of developmental disabilities. PMID- 12850510 TI - Vertebral artery insufficiency as a possible mechanism for sudden infant death- in vivo evidence does not support findings from postmortem studies. AB - Recent postmortem studies have suggested that sudden infant death syndrome (SIDS) might involve an underlying, gradual brain stem injury caused by repeated episodes of transiently compromised brain stem circulation. Autopsy studies have also reported that vertebral artery occlusion due to head rotations, such as occurs, e.g. during prone sleeping, would be a physiological phenomenon of infant atlantooccipital junction. The present study was undertaken to examine whether vertebral artery insufficiency does truly occur in live infants during such head rotations. We studied by transcranial doppler sonography the blood flow velocity of the basilar artery (BA) in 27 infants during head rotation from straight position to maximal rotation in three directions (left, right, dorsiflexion). No significant change in BA blood flow was seen between any head positions. Weight and gestational age, but not arterial pressure or hematocrit, of the infants were correlated with blood flow velocity. Our results suggest that brain stem circulation in live infants may not be compromised due to changing the head position, which is inconsistent with the postmortem findings showing insufficiency of brain stem circulation in both controls and those succumbed to SIDS. We hence propose that the brain stem pathology observed with SIDS is likely caused by other factors (e.g. systemic disturbance) rather than by mechanical obstruction of brain stem circulation. PMID- 12850512 TI - EEG patterns in 10 extreme premature neonates with normal neurological outcome: qualitative and quantitative data. AB - The aim of this prospective study was to describe and quantify EEG patterns in 10 very premature infants (24 weeks 2 days- 26 weeks 4 days GA) without neonatal neurological pathology and with a normal outcome at 3 years of age for nine of them. EEG and eye movements were recorded in the first 5 days of life. All tracings were discontinuous; EEG inactivity (<15 microV) never exceeded 1 min, representing 45.3% of total recording time. The EEG bursts, mainly synchronous, could last up to 83 s when >50 microV and to 197 s when >15 microV. High voltage delta waves (0.5 Hz; up to 330 microV), either smooth or superimposed with 7-12 Hz rhythms, were the most typical and frequent figures, mainly in temporal (mean number 257.2 +/- 73.3) and occipital (237.7 +/- 65.8 per hour recording) areas. In temporal areas, they appeared mainly in clusters; more often unilateral than bilateral (P < 0.05). Occipital delta waves were as often bilateral and synchronous as unilateral. The two distinct frontal delta waves were significantly less numerous than other delta waves (P < 0.05). Bursts of hypersynchronous high voltage delta waves and of diffuse sharp theta waves were less numerous than other waves (P < 0.01). Considering periods with or without eye movements, the mean percentage and the mean longest period of EEG activity (< or =50 microV) were significantly greater (P < 0.01) when eye movements were present, indicating a rough sleep state differentiation as early as 25 weeks CA. These EEG patterns are qualitatively and quantitatively reproducible. They constitute standards of normality and a basis for the determination of neurological prognosis. PMID- 12850511 TI - Effect of aging on autonomic function in individuals with severe motor and intellectual disabilities. AB - Autonomic nervous function is often abnormally regulated in individuals with severe motor and intellectual disabilities (SMID). In this study, we assessed autonomic nervous function of patients with SMID and determined how it was influenced by age. The study included 21 individuals with SMID and 15 healthy adolescents (control). To determine the effect of aging, the patients with SMID were divided into an older age group (Old) and younger age group (Young). Autonomic nervous function was assessed using power spectral analysis of heart rate variability (HRV) for 24-h Holter electrocardiogram recordings. The low- and high-frequency components (LH and HF) of HRV were calculated. The ratio between LF and HF (LF/HF) was used as an indicator of sympathetic modulation, while HF alone was used as an indicator of parasympathetic modulation. The LH/HF in the control group was higher in the daytime than at nighttime, while HF had an opposite pattern of change. Therefore, circadian rhythms were observed in the control group for both sympathetic and parasympathetic nervous activities. In contrast, the LF/HF in patients with SMID had no circadian rhythm. The HF was higher in the Old SMID group than in the Young group, and it exhibited a circadian rhythm in eight patients in the Old SMID group versus none of the patients in the Young group. These findings suggest that the sympathetic nervous system is suppressed in both old and young patients with SMID. However, while the parasympathetic nervous system is suppressed in younger patients with SMID, it is activated with increasing age. PMID- 12850513 TI - Activity of neurons in ventrolateral respiratory groups during swallowing in decerebrate rats. AB - To elucidate the neuronal basis of the coordination between swallowing and respiration, we examined the swallowing-related activity of respiratory neurons in the ventrolateral respiratory groups of the medulla oblongata of decerebrate, paralyzed and artificially ventilated rats (n = 14). Extracellular recording was made during fictive swallowing evoked by the electrical stimulation of the superior laryngeal nerve from a total of 141 neurons with respiratory rhythm (99 expiratory and 42 inspiratory neurons). The burst of discharge by the hypoglossal nerve was used to monitor the pharyngeal phase of swallowing. The decrementing expiratory (E-DEC) neurons (n = 62) were activated during (n = 46) or after (n = 10) the hypoglossal bursts, or showed no swallowing-related activity (n = 6). All of the augmenting-expiratory (E-AUG) neurons (n = 37) were silent during the hypoglossal bursts but were activated after each swallow. Inspiratory neurons showed either no swallowing-related bursts (n = 27), or were activated after the hypoglossal bursts (n = 15). Activation of the majority of E-DEC neurons may be related to the arrest of respiration during swallowing, and the post-swallow activation of E-AUG neurons may correspond to the expiratory phase that follows swallowing. We suggest that these behaviors of expiratory neurons are essential in the phase resetting of the respiratory cycle in association with the swallowing. PMID- 12850514 TI - The spectrum of phenotypes in females with Rett Syndrome. AB - Since the discovery of mutations in the methyl-CpG binding protein-2 (MECP2) gene in Rett Syndrome (RTT) a large number of females have been diagnosed worldwide. In this article we present the clinical and developmental data of 120 RTT females with mutations in the MECP2 gene and individually describe typical and atypical cases. We found a broad spectrum of phenotypes in females. At the severest end we have females with primary developmental delay who never learned to turn, sit or walk and who developed severe epilepsy. At the mildest end of the spectrum, there are females with only minor neurological symptoms who have good gross motor function, speak and have relatively well-preserved hand function. A number of girls either do not fulfil all the necessary diagnostic criteria or present with symptoms that have not been described in RTT before. Comparing our data with the normal population we found that girls with RTT have a smaller occipito-frontal circumference, shorter length and lower weight at birth. As a result of molecular genetic analysis a broad spectrum of phenotypes in RTT females has evolved. We found evidence that the defect in MeCP2 influences the somatic growth before birth. PMID- 12850515 TI - Re-evaluation of short latency somatosensory evoked potentials (P13, P14 and N18) for brainstem function in children who once suffered from deep coma. AB - One of the major clinical features of brain death is deep coma. Therefore, we re evaluated retrospectively electrophysiological examinations of brainstem function in about 31 children who had once suffered from deep coma in order to reveal its pathophysiological characteristics. The patient age at coma ranged from 1 month to 10 years (mean 2 years 1 month). The electrophysiological examinations were performed, including any of short-latency somatosensory evoked potential (SSEP), brainstem auditory evoked potential (BAEP) and blink reflexes. We first compared results between the fair and poor prognostic groups, and then re-evaluated SSEP results on a few severely impaired patients with persistent vegetative state (PVS). Subsequently, SSEP clarified more specific findings for a deep coma condition than BAEP and blink reflex. A lack of P14, N18 and N20, and an amplitude reduction or vagueness of P13 in SSEP in these children strongly suggested high risk in their future neurological prognosis. In conclusion, electrophysiological examinations, especially SSEP (P13, P14 and N18), might be very useful in obtaining a long-term neurological prognosis after deep coma in children. PMID- 12850516 TI - Pallister-Mosaic syndrome and neuronal migration disorder. AB - We diagnosed Pallister-Mosaic syndrome (PMS) in a 4-month-old female infant. In addition to the presence of non-specific anomalies, involving anorectal, finger and ear anomalies, characteristic cranio-facial features and irregular skin lesions that appeared after age 2 months suggested the possibility of genetic mosaicism, PMS in particular. Fluorescence in situ hybridization technique revealed an extra copy of chromosome 12p; i (12p) in 30% of cultured skin fibroblasts. When focal skin lesions accompany neurodevelopmental disabilities in early infancy, genetic analysis for mosaicism should be considered for differential diagnosis. Significantly, we describe several phenotypic features and neuroimaging findings of the PMS in the present case, which have not been described in previous reports. The neuroimaging abnormalities we encountered, such as polymicrogyria, speculating congenital brain anomaly, may explain the severe motor and intellectual disabilities of PMS. PMID- 12850517 TI - A case of megalencephalic leukoencephalopathy with subcortical cysts (van der Knaap disease): molecular genetic study. AB - Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is an autosomal recessive disorder characterized by macrocephaly, deterioration of motor function with ataxia, spasticity and mental decline. It has been revealed that the mutations in the gene, KIAA0027, were responsible for MLC and the gene was renamed subsequently 'MLC1'. A 41-year-old Japanese male with MLC, in whom a homozygous missense mutation, TCG to TTG at codon 93 resulting in S93L, was detected in the MLC1 gene, was described. MRI revealed marked cerebral atrophy and enlargement of the ventricular system. The subject's motor function had severely deteriorated, while his cognitive function had maintained at the level of a 2-year-old for the past 10 years. The mutation in the MLC1 gene of the patient is considered to be a common mutation responsible for MLC in Japanese patients because the same mutation had been detected in two other Japanese patients with MLC. PMID- 12850518 TI - Combination therapy with intraventricular interferon-alpha and ribavirin for subacute sclerosing panencephalitis and monitoring measles virus RNA by quantitative PCR assay. AB - Subacute sclerosing panencephalitis (SSPE) is a degenerative disease of the central nervous system that leads to death within a few years. Recently, it has been reported that combination therapy with intraparenchymal interferon-alpha (INF-alpha) and intraventricular ribavirin is effective. An 11-year-old SSPE patient whose clinical symptoms progressed rapidly, was treated first with intraventricular INF-alpha and then with combined intraventricular INF-alpha and ribavirin therapy. To monitor viral load over the course of the therapy, measles virus RNA was quantified using a real-time polymerase chain reaction assay. Measles virus RNA decreased rapidly after the INF-alpha therapy was started, paralleling the decrease in the measles antibody titer in the cerebrospinal fluid and the improvement in the neurological disability. After intraventricular ribavirin was combined with INF-alpha therapy, no further improvement was observed. The neurological disability gradually progressed, although the amount of virus RNA remained low. PMID- 12850522 TI - Epithelial cell differentiation in the human prostate epithelium: implications for the pathogenesis and therapy of prostate cancer. AB - Within the human prostate epithelium four cell populations are discriminated by their expression of keratins (K). While basal cells co-localize K5 and K14 combined with low levels of K18 (K5(++)/14(++)/18(+)), luminal cells highly express K18 (K18(++)). In addition, two intermediate subpopulations are characterized either by basal K5(++)/18(+)- or luminal K5(+)/18(++)- expression. The entire prostate epithelium is putatively derived from a basal stem cell population. They give rise to intermediate cells that transiently proliferate and mature towards differentiated luminal epithelium. Within prostate carcinoma luminal exocrine, neuro-endocrine and intermediate cells are distinguished. Intermediate cells have been postulated as progenitors for prostate carcinogenesis and targets for androgen-independent tumor progression. Androgen independency is associated with an enrichment of intermediate cells and over expression of peptide growth factor receptors. Targeting intermediate cells by inhibition of their peptide growth factor receptors, therefore, offers novel treatment modalities for prostate cancer. PMID- 12850523 TI - Searching for the gatekeeper oncogene of prostate cancer. AB - Prostate cancer is a common malignancy that has a heterogeneous etiology and a variable outcome. Nearly all prostatic adenocarcinoma results from androgen dependent tumor promotion. However, the cause of prostate cancer initiation is not well understood and only a few of the target oncogenes activated during prostate cancer initiation have been identified. Prostate cancer risk is strongly influenced by family history. Several genetic loci have been found to cosegregate with prostate cancer occurrence in high-risk families. Some candidate oncogenes that map to these loci have been implicated by the identification of mutations in high-risk kindreds. However, the roles of the putative oncogene products in the biochemical pathways that mediate carcinogenesis remain obscure and their influence on cancer etiology has yet to be supported by gene targeting experiments in mice. Moreover, the genes that have been implicated in hereditary prostate cancers do not appear to be mutated in sporadic cancers. Karyotypic and loss of heterozygosity analysis of sporadic prostate cancers have identified 8p, 10q, and 17p as the loci most often disrupted. Candidate oncogenes have been identified at each of these regions. Additional genes with pathogenic significance in prostate cancer have been identified by analysis of cDNA microarrays comparing benign and malignant prostate tissue, by differential genetic analysis of benign and malignant prostatic epithelium, and by induction of experimental prostate cancer in genetically engineered mice. PMID- 12850524 TI - Optimizing targeted therapy and developing novel outcome measures for patients with advanced prostate cancer at Memorial Sloan-Kettering Cancer Center. AB - Hormonal therapy and chemotherapy, though active treatments for prostate cancer, are not curative for patients with metastatic disease. Targeted therapy has the potential to control, if not eradicate, cells resistant to castration and chemotherapy. Despite several years of development, however, a biologic approach with clear clinical benefits has yet to emerge from a crowded field. This review outlines the approaches being studied at Memorial Sloan-Kettering Cancer Center to optimize biologic therapy. Trials of targeted therapy are designed on the basis of a clinical states model that describes both patient clinical risks and tumor biology. Drugs that act on multiple pathways are being developed, and targets that are expressed across all phases of the disease are selected. New molecular imaging techniques permit assessments of the target before, during, and after treatment. High-throughput preclinical assays of gene expression are being developed to enhance selection of drug sequences and combinations for clinical testing. PMID- 12850525 TI - Prognostic factors and clinical trials of new agents in patients with metastatic renal cell carcinoma. AB - Metastatic renal cell carcinoma (RCC) remains a disease highly resistant to systemic therapy. Results of recent reports in the literature on prognostic factors and clinical trials for patients with metastatic RCC were reviewed. Small numbers of patients exhibit complete or partial responses to interferon and/or interleukin-2, but most patients do not respond and there are few long-term survivors. Therefore, the identification of new agents with better antitumor activity against metastases remains the highest priority of clinical investigation in this refractory tumor. Prospective identification of patients more likely to benefit from cytokine therapy could be used as a stratification factor in Phase III trials, and in risk-directed therapy. PMID- 12850526 TI - Angiogenesis inhibitors in genitourinary cancers. AB - Despite much enthusiasm, no clear clinical benefit to any antiangiogenic agent has yet been demonstrated. Phase I trials demonstrate that endostatin, an endothelial cell toxin, can be administered safely, but no obvious anti-tumor effects were observed. Certain matrix metalloproteinase inhibitors appear ineffective, but later generation inhibitors with less systemic toxicity continue to be investigated. There are occasional responses to thalidomide either singly or in combination, but its pharmacology and mechanism of action remain unclear. A randomized study with the anti-VEGF antibody bevacizumab suggests that VEGF pathway is an important target in renal cell cancer. VEGF receptor tyrosine kinase inhibitors continue to be developed, but one of the first compounds SU5416 has had minimal clinical effects. Clinical trial designs that address the stable disease endpoint should thus be investigated and the randomized discontinuation design has already been tested. Pharmacodynamic markers that reflect antiangiogenic drug effect also need to be developed, but the putative ones, including circulating proangiogenic factors, tumor microvessel density, and dynamic contrast MRI have not yet proven to be useful. PMID- 12850527 TI - Current status of thalidomide and its role in the treatment of metastatic prostate cancer. AB - Following the discovery of its anti-angiogenic properties and despite its tragic history, thalidomide has re-surfaced in the field of oncology. Concurrent with its evaluation in various clinical trials for cancer, thalidomide's mechanism of action is sought and new analogues with improved efficacy and pharmacological profile are emerging. This review is a critical evaluation of thalidomide metabolism, molecular targets, anti-angiogenic activity and clinical efficacy with an emphasis on metastatic prostate cancer. PMID- 12850528 TI - Thalidomide therapy for renal cell carcinoma. AB - PURPOSE: To review the use of thalidomide as a therapeutic option for patients with metastatic renal cell carcinoma (RCC). MATERIALS AND METHODS: Studies of thalidomide alone or in combination with immunotherapy or chemotherapy were identified. The clinical benefit of thalidomide was assessed on the basis of objective response (complete and partial) and stable disease rates. RESULTS: Single-agent thalidomide was evaluated in nine phase II studies, producing partial responses in a median of 7% of patients (range: 0-17%) and stable disease in a median of 31% of patients. On average, 40-45% of patients derived benefit from thalidomide. Common toxicities included constipation, lethargy, and with prolonged therapy, neuropathy. Four studies reported deep vein thrombosis and/or pulmonary embolism at rates ranging from 3 to 23%. On the basis of these findings, phase I/II studies of thalidomide in combination with interleukin-2 (IL 2), interferon (IFN), or chemotherapy have been conducted. Although some of these early combination regimens were limited by toxicity, promising findings have been seen with thalidomide/IL-2 and thalidomide/IFN/capecitabine. A phase III trial of IFN versus IFN/thalidomide is nearing completion. CONCLUSION: Thalidomide is active in metastatic RCC, but additional experience with thalidomide-based combinations is needed to better define how this agent should be used in the management of this malignancy. PMID- 12850529 TI - Therapeutic approaches to bladder cancer: identifying targets and mechanisms. AB - Transitional cell carcinoma is the second most common genitourinary malignancy in US and third most common cause of death among genitourinary tumors. Treatment options for bladder cancer include surgery, often combined with chemotherapy, radiation, and/or immunotherapy. The MVAC adjuvant chemotherapy regimen has been most widely used in locally invasive as well as metastatic disease. Only a proportion of patients at risk will respond to therapy. There is thus need to identify good responder patients for adjuvant therapy and to identify new targets to treat a greater range of patients. Based upon patient-specific aberrations in pathways or known markers, both existing and new therapies can be tailored to benefit patients based on the risk of progression and molecular alterations specific to a patient's tumor. Targeted therapy, therefore, is defined as therapy that targets mechanism and risk. Utilizing the available knowledge of the molecular biology of cell-cycle regulation, signal transduction, apoptosis, and angiogenesis in bladder cancer, we review the potential therapeutic targets for rational drug development. Finally, using bladder cancer as a model for translational research, requirements for a desired clinical trial are presented. PMID- 12850530 TI - Novel approaches with targeted therapies in bladder cancer. Therapy of bladder cancer by blockade of the epidermal growth factor receptor family. AB - The improved understanding of the molecular biology of urothelial malignancies is helping to define the role of new targets and prognostic indices that can direct the most appropriate choice of treatment for advanced disease. Many human tumors express high levels of growth factors and their receptors that can be used as potential therapeutical targets. Tyrosine-kinase receptors, including many growth factor receptors such the receptors for epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), and Her2/neu, have been found overexpressed in urothelial tumors. For many of these growth factor receptors, the degree of expression has been associated with the progression of cancer and a poor prognosis. Among the best studied growth factor receptors are the two members of EGF receptor familiy EGFr (ErbB-1), and Her2/neu (ErbB-2). Several preclinical studies in bladder cancer models, have confirmed that systemic administration of growth factor inhibitors inhibits the growth and metastasis of human transitional cell carcinoma established in the bladder wall of athymic nude mice. Additional studies indicate that therapy with EGFR inhibitors enhances the activity of conventional cytoreductive chemotherapeutic agents, in part by inhibiting tumor cell proliferation, angiogenesis, and inducing apoptosis. Novel targeted therapy hold promise to improve the current results of bladder cancer treatment. Based on the success seen with anti-HER2 monoclonal antibodies (Herceptin) and the promising results with EGFR targeted agents (IMC-C225 Cetuximab, ZD1389 Iressa, OSI-774 Tarceva, GW 57016) in other tumor types, and based on the results obtained in preclinical models, there is a great interest in assessing these agents in patients with bladder cancer. Several trials are now ongoing testing these new agents alone or in combination with chemotherapy in bladder cancer patients. The integration of these newer biologic agents, probably to supplement rather than to supplant chemotherapeutic drugs, should be a primary direction of research with the objective to interfere with multiple aspects of bladder cancer progression. However, the value of integration of biologically targeted agents into combined modality treatment for patients with bladder cancer has still to be proven. PMID- 12850531 TI - Gemcitabine, paclitaxel, pemetrexed and other newer agents in urothelial and kidney cancers. AB - Significant progress has been made in understanding the biology of urothelial and kidney cancers. Approaches to advanced urothelial cancer include dose intensification, reducing toxicity in unfit or elderly patients, doublet and triplet combination chemotherapy and sequential regimens. Promising new chemotherapeutic agents such as the epothilones, pemetrexed (Alimta), topoisomerase inhibitors and vinflunine act at different phases of the cell cycle and on folate metabolism. New agents that are combined with chemotherapy in urothelial cancer include the farnesyl transferase inhibitors and growth factors receptor inhibitors. Renal cell carcinoma (RCC) is particularly resistant to cytotoxic agents, although a gemcitabine/fluorinated pyrimidine combination may have modest but real clinical benefit. In metastatic RCC, new biologic and targeted therapies include anti-angiogenesis agents such as anti-vascular endothelial growth factor (VEGF) antibody and thalidomide, as well as toremifene, CCI-779 and allogeneic stem cell transplantation. Metastatic urothelial and renal cell cancers continue to be the clinical trial focus of many novel agents. The molecular biology of these diseases is being unravelled and as knowledge accumulates, our ability to target these cancers will continue to increase. PMID- 12850532 TI - The potential for prostate cancer immunotherapy. AB - The components of an effective immune response have been elucidated in recent years. An understanding of the dysfunctional immune response in cancer in one or more of these components has lead to a variety of immunotherapeutic approaches. These therapeutic strategies are designed to stimulate dendritic cell (DC) proliferation, promote antigen uptake and processing, stimulate an effector cell response via direct antigen presentation or target tumor cells via antibody therapy. Many approaches in prostate cancer have demonstrated successful induction of the desired immune response. Limited clinical success has also been seen. PMID- 12850533 TI - Selective A2A, but not A1 adenosine antagonists enhance the anticataleptic action of trihexyphenidyl in rats. AB - In rats made cataleptic with haloperidol (5.32 micromol/kg), the bar test was used to assess the possible synergism between the muscarinic antagonist trihexyphenidyl (THP) and selective adenosine A(1) and A(2A) receptor antagonists. Neither catalepsy intensity nor latency were affected by a subthreshold dose of THP (0.33 micromol/kg). The selective adenosine A(1) antagonist 8-cyclopentyl-1,3-dipropyl-xanthine (DPCPX) (5.15 micromol/kg) caused a small, but significant reduction of catalepsy intensity that remained unchanged when combined with THP. DPCPX had no effect on catalepsy latency, either alone or combined with THP. In contrast, an equimolar dose of the selective adenosine A(2A) antagonist 4-(2-[7-amino-2-(2-furyl)1,2,4-triazolo[2,3-a]-[1,3,5]triazin-5 ylamino]ethyl)phenol (ZM 241385) (5.15 micromol/kg) produced a significant reduction of catalepsy intensity and increased catalepsy latency. Both effects were potentiated when ZM 241385 was combined with THP. The synergism was more evident when rats were pretreated with a subthreshold dose of ZM 241385 (1.55 micromol/kg) that was unable to modify catalepsy parameters when applied alone, but produced a significant reduction in catalepsy intensity and an increase in catalepsy latency when administered with THP. Catalepsy was unaffected by a combination of equimolar, subthreshold doses of DPCPX (1.55 micromol/kg) and ZM 241385 (1.55 micromol/kg). These findings indicate that the anticataleptic effect of anticholinergics is enhanced only by the selective blockade of adenosine A(2A) receptors. PMID- 12850534 TI - The anxiogenic beta-carboline FG-7142 inhibits locomotor exploration similarly in postweanling and adult rats. AB - The beta-carboline FG-7142 exerts many stress-like effects in the adult rat, including the inhibition of locomotor exploration. However, comparable effects in immature animals have not been reported. Stress-like effects of FG-7142 are mediated via its inverse agonist actions on gamma-aminobutyric acid (GABA) receptors. GABA systems change considerably with development and GABA agonists such as diazepam have opposite behavioral actions in immature and adult rats. Accordingly, we compared FG-7142 effects on locomotor exploration in postweanling and adult rats. Postweanling male rats (24 days) and adult rats (90 days) received a single injection of vehicle or FG-7142 (15 mg/kg i.p.) and 15 min later were placed in photocell monitors for 1 h. Although postweanling animals traveled a smaller distance overall than adults, FG-7142 inhibited locomotor exploration to a similar degree in both groups. We conclude that FG-7142 exerts stress-like effects in postweanling rats and may be considered for use as a model of childhood stress. PMID- 12850535 TI - Differentiation-inducing and protective effects of adult rat olfactory ensheathing cell conditioned medium on PC12 cells. AB - Transplantation of olfactory ensheathing cells has been one of the promising strategies in enhancing central nerve fiber regeneration. Membrane surface molecules on olfactory ensheathing cells mediating cell-cell interactions as well as various factors released from them are thought to be important for nerve regeneration. The latter, however, has not been fully substantiated by experimental data, particularly regarding the olfactory ensheathing cells of the adult animals. In the present study, the effects of adult olfactory ensheathing cell conditioned medium on PC12 cells were examined. The results show that the factors secreted by the adult olfactory ensheathing cells can promote PC12 cell differentiation and protect it from Zn(2+) insult. PMID- 12850536 TI - Involvement of beta-arrestin-2 in modulation of the spinal antinociception induced by mu-opioid receptor agonists in the mouse. AB - Beta-arrestins have been suggested to regulate mu-, delta-, and kappa-opioid receptor-mediated responses. In the present study, we examined the effects of pretreatment with beta-arrestin-2 antibody on tail-flick inhibition induced by opioid receptor agonists in the mouse spinal cord. Intrathecal (i.t.) pretreatment with beta-arrestin-2 antibody potentiated the antinociception induced by i.t.-administered mu-opioid receptor agonists [D-Ala(2),NMePhe(4),Gly ol(5)]enkephalin (DAMGO) and endomorphin-1, but not endomorphin-2, the delta opioid receptor agonist [D-Ala(2)]deltorphin II or the kappa-opioid receptor agonist U50,488H. The present result suggests that beta-arrestin-2 may tonically down-regulate a selected population of mu-opioid receptors activated by endomorphin-1 or DAMGO in the mouse spinal cord. PMID- 12850537 TI - Contribution of ionotropic glutamate receptors to acute amphetamine-stimulated preproenkephalin mRNA expression in the rat striatum in vivo. AB - The present study evaluated the possible role of ionotropic glutamate receptors in mediating the inducible preproenkephalin (PPE) mRNA expression in the rat striatum in response to systemic administration of a psychostimulant amphetamine. A single injection of amphetamine (5 mg/kg, i.p.) increased PPE mRNA levels in the dorsal striatum as revealed via quantitative in situ hybridization. Pretreatment with the NMDA receptor antagonist, MK801 (0.5 mg/kg, i.p.) or 3-((R) 2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (10 mg/kg, i.p.), attenuated the amphetamine-induced PPE mRNA expression. A similar attenuation of the amphetamine-induced PPE expression, although to a lesser extent, was also observed following pretreatment with a kainate/AMPA receptor antagonist, 6,7 dinitroquinoxaline-2,3-dione (100-200 mg/kg, i.p.). Three antagonists alone had no significant effect on basal levels of PPE mRNAs in the dorsal striatum. These data indicate that both NMDA and kainate/AMPA receptors are involved in coupling extracellular signals evoked by acute stimulant exposure to PPE mRNA expression in striatal neurons. PMID- 12850538 TI - Role of nitric oxide in lipopolysaccharide-induced release of vasopressin in rats. AB - This study evaluated the role of nitric oxide (NO) in vasopressin (AVP) release induced by intravenous lipopolysaccharide (LPS) in rats previously treated with intracerebroventricular (i.c.v.) saline, L-NAME, L-arginine or sodium nitroprusside (SNP). In control rats given i.c.v. saline, L-NAME, L-arginine or SNP, AVP levels did not change from baseline. After LPS, plasma AVP increased, reaching a peak at 60 min, and returning to basal levels 4 h later in all i.c.v. pre-treated groups (P<0.05). The LPS administration in rats previously treated with L-NAME induced higher AVP levels (P<0.05) that remained elevated throughout the period of the experiment (P<0.05). These findings confirm the inhibitory role of NO in AVP secretion induced by LPS. PMID- 12850539 TI - Reproductive status influences the survival of new cells in the dentate gyrus of adult male meadow voles. AB - Reproductive status influences cell proliferation and the survival of new cells in the dentate gyrus of adult laboratory-reared and wild female meadow voles; reproductively inactive (RI) females have more proliferating cells and more labeled cells that survive 5 weeks vs. 2 h than reproductively active (RA) females. However, the effect of season has only been studied in a wild sample of male meadow voles and factors such as age and experience that have been shown to influence neurogenesis in the dentate gyrus of adult mammals cannot be controlled in a wild sample. Therefore, we investigated whether reproductive status regulates neurogenesis (cell proliferation and/or the survival of new cells) in the dentate gyrus of laboratory-reared adult male meadow voles so that confounding variables could be controlled. Males were acclimated to a short- or a long-photoperiod to simulate the non-breeding or breeding season, respectively, and reproductive status was verified by testes mass. The density of labeled cells was similar in RI and RA males (P=0.20) 2 h after an injection of bromodeoxyuridine (50 mg/kg) but the density of labeled cells was elevated in the RA males relative to the RI males 5 weeks after an injection of [(3)H]thymidine (5 microCi/g; P or =3/10 was defined as clinically significant fatigue. Possible independent variables were identified from a biopsychosocial model of fatigue. Fisher's exact test was used to univariately assess the association of each variable with clinically significant fatigue. Multiple logistic regression analyses were used to identify independent predictors of fatigue within each dimension, and then across dimensions. Fatigue was present in 113 (62%) patients, and 80 (44.4%) patients had usual fatigue > or =3/10. The unidimensional independent predictors were use of analgesics (situation dimension); hemoglobin and serum sodium (biomedical dimension); feeling drowsy, dyspnea, pain and lack of appetite (physical symptom dimension); and feeling sad and feeling irritable (psychological symptom dimension). In a multidimensional model, dyspnea, pain, lack of appetite, feeling drowsy, feeling sad, and feeling irritable predicted fatigue independently with good calibration (Hosmer Lemeshow Chi Square=5.73, P=0.68) and discrimination (area under the receiver operating characteristic curve=0.88). Physical and psychological symptoms predict fatigue independently in the multidimensional model, and superseded laboratory data. These findings support a symptom-oriented approach to assessment of cancer-related fatigue. PMID- 12850644 TI - Identifying ambulatory cancer patients at risk of impaired capacity to consent to research. AB - Ethicists and others have expressed concerns that some patients with cancer might lack adequate decision-making capacity to give consent for research. Although this concern is plausible, it is not known what patient characteristics might be used to identify those patients who are at risk and who therefore should undergo a formal assessment of decision-making capacity. Forty-five patients with cancer were presented with a description of a randomized controlled trial, accompanied by an Institutional Review Board-approved consent form. Two raters who were blind to all patient characteristics assessed decision-making capacity using the MacArthur Competency Assessment Tool for Clinical Research. These scores were summarized in overall capacity judgments using criteria established by a panel of experts. Subjects also completed a symptom rating scale and a battery of neuropsychiatric tests. No relationship was observed between symptom severity and any domain of decision-making capacity (understanding, appreciation, reasoning, ability to express a choice) or summary judgments. However, several other patient characteristics, including age, education, and selected neuropsychiatric test results, were found to be strongly associated with capacity scores. These data suggest that several patient characteristics, such as age, education, and tests of cognitive functions, may help investigators to identify patients with impaired capacity to give consent for research. PMID- 12850645 TI - Old people in pain: a comparative study. AB - To investigate the prevalence of pain in older people (75+), compare those in pain to those without regarding demographics, social network, functional limitations, fatigue, sleeping problems, depressed mood and quality of life (QOL), and identify variables associated with pain, a cross-sectional, prospective survey was conducted in an age-stratified sample of 4,093 people aged 75-105 years old. Those reporting pain (n=1,654) were compared with those who did not (n=2,439). Pain was more common with higher age, as were all complaints among those in pain and among those without, except sleeping problems. Lower QOL was found with higher age, as well as with pain. Pain was found to be associated with functional limitations, fatigue, sleeping problems, depressed mood, and QOL. These data highlight the importance of identifying old people in pain. Those who are older and those affected by pain are at greater risk of also being troubled by other problems, such as functional limitations and lowered QOL. PMID- 12850646 TI - Place of death of Houston area residents with cancer over a two-year period. AB - The majority of cancer patients wish to die at home. Improved understanding of place of death and its relevant demographic predictors is important for the planning of palliative cancer care programs. The purpose of this study was to determine the place and predictors of site of death in cancer patients in a major U.S. metropolitan area. Death certificate data over two years were analyzed for Houston area residents with cancer who died in the Houston area. Information was obtained on factors that might be associated with the place where cancer patients die. For the purpose of this study, we looked at the following variables: primary site of cancer (hematological, breast, genitourinary, gastrointestinal, lung, and other); black, white, Hispanic, or Asian; age at death; marital status; sex; whether or not veteran of U.S. armed forces; levels of education; and area of residency within the Houston area. Univariate and multivariate analyses were performed. The majority of patients died in the hospital (51-52% both years), with the next most frequently occurring group dying at home (34-35% both years). Stepwise multivariate analysis resulted in a 6-variable logistic regression model. In this model, the odds of dying in hospital were increased by a factor of 2.7 if the patient had a hematological cancer (P<0.0001), a factor of 1.6 if the patient lived in Harris County (P<0.0001), and a factor of 1.5 if the patient was black (P<0.0001). Further characterization of factors associated with increased risk of hospital death rate is needed and systems should be developed to enable the majority of cancer patients to access palliative care services in the multiple settings in which they die. PMID- 12850647 TI - The memorial symptom assessment scale: modified for use in understanding family caregivers' perceptions of cancer patients' symptom experiences. AB - The purpose of this study was to determine whether the Memorial Symptom Assessment Scale (MSAS) could serve as a feasible, reliable, and valid tool for use in assessing the reports of 98 family caregivers (FC) on the symptom experiences of advanced stage cancer patients. The MSAS consists of subscales that describe psychological symptom distress (PSYCH), physical symptom distress (PHYS), and global symptom distress (Global Distress Index [MSAS-GDI]). The majority of FCs was able to respond to 32 MSAS symptom items; however, the greatest difficulty in FC responding occurred on the 'sexual interest' item. The mean PSYCH score was 1.42 (SD=0.83), the mean PHYS score was 0.96 (SD=0.67), and the mean MSAS-GDI was 1.46 (SD=0.24). Internal consistency was high in the PHYS (alpha=0.84), PSYCH (alpha=0.82), and MSAS-GDI (alpha=0.84) subscales. The average item-scale correlation ranged between r=0.50 and 0.60, indicating that the items were moderately to strongly correlated with the respective total subscales. The average inter-item correlation ranged between r=0.30 and 0.45, indicating that the items were moderately correlated with each other on the respective subscales. Good to excellent intraclass correlations (ICC) with patients' ordinal ratings support the concurrent validity and utility of the PHYS and MSAS-GDI subscales in FC populations who care for cancer patients in the home setting. Depending on the symptom, a range of slight to substantial Kappa values on dichotomous PHYS and PSYCH items, and fair ICC values on the ordinal PSYCH subscale provide questionable validity in FC populations caring for cancer patients. PMID- 12850648 TI - Definitions related to the medical use of opioids: evolution towards universal agreement. AB - Misunderstandings regarding the nature and occurrence of addiction have historically been barriers to the appropriate treatment of pain and have stigmatized the medical use of opioids. This article reviews the evolution of nomenclature related to addiction, presents current scientific understanding of addiction that may help shape universally acceptable terminology, and discusses an integrated effort of pain and addiction professionals to reach consensus on addiction-related terms. The article suggests key principles that may clarify terminology including: clear differentiation of the concepts of addiction and physical dependence, conceptualization of addiction as a multidimensional disease, and use of a label for the phenomenon of addiction that does not include the ambiguous term "dependence." More universal agreement on terminology related to addiction is expected to improve the treatment of both pain and addictive disorders; improve communication between health care providers, regulators, and enforcement agencies; and reduce health care and other societal costs. PMID- 12850649 TI - Long-term outcomes during treatment of chronic pain with intrathecal clonidine or clonidine/opioid combinations. AB - To evaluate the effectiveness of intrathecal clonidine or clonidine/opioid admixture for the treatment of chronic pain states, a retrospective chart audit of 15 patients seen by the Pain Medicine and Neurosurgical Services was performed. Subjects included 9 men and 6 women aged 26-86 years. Diagnoses included complex regional pain syndrome, neuropathic pain, and cancer pain. All patients received a trial of single-shot and/or short-term infusion of clonidine. Those reporting a significant reduction in pain, or at least 50% reduction in their visual analog scale (VAS), received long-term therapy. Intrathecal clonidine as a single-shot dose, infusion, or as intrathecal polytherapy did not improve VAS scales from pre-treatment values in 5 patients. Ten patients reported significant pain relief or >50% decrease in VAS scores with the initial trial and received long-term therapy. Two received clonidine alone for 7-11 months before the therapy failed; others failed after just a few days. Seven of eight initially responded to clonidine alone (75-950 microg/day) before failing and requiring a second drug. Three received hydromorphone (200-8000 microg/day) and four morphine (0.15-15 mg/day) with clonidine. Four patients then failed 2-drug therapy (duration 6-21 months). Two continue with intrathecal clonidine/hydromorphone (duration 19-29 months) and 1 with clonidine/morphine (duration 21 months). After initiation of intrathecal clonidine, one patient reported good relief with clonidine/morphine until his death 5 months later. In this population, intrathecal clonidine was of limited utility for most patients. It may be of benefit for subset(s) of patients, but in our experience, duration of relief is typically <18 months. PMID- 12850650 TI - Treatment of chronic mechanical spinal pain with intravenous pamidronate: a review of medical records. AB - We explored the effect of intravenous infusions of a bisphosphonate, pamidronate, in the management of chronic mechanical spinal pain, a worldwide public health problem in terms of lost workdays, medical treatment costs, and suffering. Bisphosphonates have an anti-nociceptive effect in animals. In humans, intravenous pamidronate relieves numerous painful conditions, including metastatic bone pain, ankylosing spondylitis, rheumatoid arthritis, and complex regional pain syndrome. We reviewed the charts of 25 patients who had experienced disabling spinal pain for several years, and whom we treated with intravenous pamidronate. None had a history of osteoporotic vertebral fractures or metastatic disease. Pain rating scores decreased in 91% of patients: on a 0-10 numeric rating scale, the mean pain change was -3.6 points and mean percentage change was -41% (P<0.0001). There was no increase in opioid or nonopioid analgesic medications associated with pain relief. The apparent analgesic effect of pamidronate for chronic mechanical spinal pain needs to be confirmed with placebo controlled trials. PMID- 12850651 TI - Anticholinergic agents for the treatment of "death rattle" in patients with myasthenia gravis. AB - Difficulty clearing upper airway secretions (death rattle) is a frequent problem at the end of life. Treatment often includes the use of anticholinergic drugs. Myasthenia gravis is a disease characterized by muscle weakness and fatigue caused by an immune-mediated deficiency of acetylcholine receptors at the neuromuscular junction, and it is treated with anticholinesterase agents. We report the case of a patient dying of myasthenia gravis who had problems with the "death rattle" and who presented a dilemma as to whether the use of anticholinergics would be helpful or would cause deterioration of her myasthenia. This is accompanied by a review of the relevant literature. PMID- 12850653 TI - The Institute of Medicine "Chasm" report: implications for depression collaborative care models. AB - The Institute of Medicine report "Crossing the Quality Chasm: A New Health Care System for the 21st Century" describes the barriers in the American health care system to improving quality of care and outcomes of chronic illness. This article describes how depression collaborative care models as well as newer research aimed at organizational and economic issues have addressed these barriers in order to improve outcomes for patients with depression in primary care systems. PMID- 12850654 TI - Measuring adherence to depression treatment guidelines in a VA primary care clinic. AB - The primary objectives of this pilot study were to develop a measure of adherence for depression practice guidelines and to assess the degree to which providers and patients adhere to guidelines in a VA primary care setting. The Depression Guideline Measure (DGM) is based on three national guidelines. The DGM was used to review medical records of 111 patients with Patient Health Questionnaire (PHQ) scores >or=10. Interrater reliability for 15 of 19 DGM checklist items was excellent (kappa > 0.75). There was a broad range of adherence to guideline criteria: only 13.5% of patients were contacted for follow-up within 2 weeks, while 100% of providers documented follow-up plans. Forty percent of patients saw mental health providers, and 63% were prescribed antidepressants. A secondary objective of the study was to explore the relationship between guideline adherence and changes in PHQ scores. Among 51 patients who completed follow-up PHQs, no associations were detected. The results suggest that the DGM shows promise as a reliable measure of guideline adherence and that guideline criteria are frequently not met in busy primary care clinics. More research is needed to determine the validity of the DGM and the impact of guideline adherence on depression outcomes. PMID- 12850655 TI - Implementing a disease management intervention for depression in primary care: a random work sampling study. AB - We describe the daily work activities of 13 Depression Clinical Specialists (DCSs) at 7 national sites who served as care managers in an effective multisite randomized trial of a disease management model for depression in primary care. DCSs carried portable random-reminder beepers for a total of 147 consecutive workdays and recorded 4,030 work activities. Patient care activity comprised the largest percentage of the workday, 49.4% (95% confidence interval [CI], 42.0 to 56.7%), followed by research-related activity, 18.3 % (95% CI, 14.7 to 21.9%), administrative work, 17.9% (95% CI, 12.2 to 23.7%), personal time, 9.4% (95% CI, 5.4 to 13.4%), and time in transit, 5.1% (95% CI, 2.8 to 7.4%). The DCSs delivered 19.2% (95% CI, 14.4 to 24.1%) of direct patient care by telephone. The DCSs spent a significant portion of the day alone 48.7% (95% CI, 43.3 to 54.1%), followed by time spent with patients, 37.5% (95% CI, 31.6 to 43.3%). Less than 10% (7.8%) (95% CI, 5.1 to 10.6%) of their time was spent with local study staff. Less than 4% of their time was spent with other health care providers. Our results demonstrate that the DCSs' time was primarily devoted to clinical care, a significant portion of which was delivered by telephone. They functioned independently, making efficient use of the limited amount of time that they interacted with other health care providers. This information will be helpful to those who may wish to implement this disease management strategy. PMID- 12850656 TI - The relationship of depressive symptoms to symptom reporting, self-care and glucose control in diabetes. AB - Depressive symptoms are common among patients with diabetes and may have a significant impact on self-management and health outcomes. In this study we predicted that: 1) there would be a significant association between depressive symptoms and diabetes symptom burden, physical functioning, diabetes self-care, and HbA1c levels; and, 2) that the association between depressive symptoms and HbA1c levels would be significantly greater in type 1, as compared to type 2 diabetic patients. This cross-sectional observational study of 276 type 1 and 199 type 2 diabetes patients took place in a tertiary care specialty clinic. We collected self-reported data on depressive symptoms, complications, medical comorbidity, diabetes symptoms, diabetes self-care behaviors, physical functioning, and demographics. From automated data we determined mean HbA1c levels over the prior year. We performed linear regression analyses to assess the association between depressive symptoms and diabetes symptom perception, diabetes self-care behaviors, physical functioning, and glycemic control. Among patients with type 1 and 2 diabetes, depressive symptoms were associated with greater diabetes symptom reporting, poorer physical functioning, and less adherence to exercise regimens and diet. There was a significant association between depressive symptoms and HbA1c levels in type 1, but not type 2 diabetic patients. Because of their association with clinical aspects of diabetes care such as diabetes symptom reporting and adherence to diabetes self-care, depressive symptoms are important to recognize in treating patients with diabetes. PMID- 12850657 TI - Psychiatric morbidity and posttraumatic symptoms among earthquake victims in primary care clinics. AB - Three months after the devastating Chi-Chi earthquake (magnitude of 7.3 on the Richter scale) struck the central area of Taiwan, 663 victims were screened for psychiatric morbidity at a local general hospital in a community mental health program. The rate of psychiatric morbidity as defined by the 12-item Chinese Health Questionnaire as greater than 4, was 24.5%. Posttraumatic symptoms were still prevalent. The rate of posttraumatic stress disorder was 11.3%, and the rate of partial PTSD was 32.0%. Variables associated with the presence of psychiatric morbidity and posttraumatic symptoms included female gender, old age, financial loss, obsessive trait, and nervous trait. A disproportionate use of mental health services (18%) was found, suggesting an urgent need to deliver mental health care to disaster victims at local medical settings. In addition, health care professionals who work with the earthquake victims need to be promptly and efficiently trained in mental health crisis intervention. PMID- 12850658 TI - Expansion of the consultation-liaison psychiatry paradigm at a university medical center: integration of diversified clinical and funding models. AB - The perspective of the contemporary Consultation-Liason Service (CLS) psychiatrist is increasingly one of consultant to medical and surgical colleagues in models other than inpatient medical and surgical units. Simultaneously, the need for a clinically and educationally robust inpatient CLS persists despite funding pressures. The University of California, Davis Medical Center Department of Psychiatry has made use of creative organizational and financial models to accomplish the inpatient CLS clinical and educational missions in a fiscally responsible manner. In addition, the department has in recent years expanded the delivery of psychiatry consultation-liaison clinical and educational services to other models of care delivery, broadening the role and influence of the CLS. Several of the initiatives described in this paper parallel an overall evolution of the practice of consultation-liaison psychiatry in response to managed care influences and other systems pressures. This consultation-liaison paradigm expansion with diversified sources of funding support facilitates the development of consultation-liaison psychiatry along additional clinical, administrative, research, and educational dimensions. Other university medical centers may consider adaptation of some of the initiatives described here to their institutions. PMID- 12850659 TI - Incidence and correlates of near-death experiences in a cardiac care unit. AB - Near-death experiences, unusual experiences during a close brush with death, may precipitate pervasive attitudinal and behavior changes. The incidence and psychological correlates of such experiences, and their association with proximity to death, are unclear. We conducted a 30-month survey to identify near death experiences in a tertiary care center cardiac inpatient service. In a consecutive sample of 1595 patients admitted to the cardiac inpatient service (mean age 63 years, 61% male), of whom 7% were admitted with cardiac arrest, patients who described near-death experiences were matched with comparison patients on diagnosis, gender, and age. Near-death experiences were reported by 10% of patients with cardiac arrest and 1% of other cardiac patients (P<.001). Near-death experiencers were younger than other patients (P=.001), were more likely to have lost consciousness (P<.001) and to report prior purportedly paranormal experiences (P=.009), and had greater approach-oriented death acceptance (P=.01). Near-death experiencers and comparison patients did not differ in sociodemographic variables, social support, quality of life, acceptance of their illness, cognitive function, capacity for physical activities, degree of cardiac dysfunction, objective proximity to death, or coronary prognosis. PMID- 12850660 TI - A validation study of the hospital anxiety and depression scale (HADS) in a Spanish population. AB - The present study aims to validate the Spanish version of the Hospital Anxiety and Depression Scale (HADS) and to determine the use of this tool for screening mood and anxiety disorders. Psychometric properties of the HADS were assessed in different groups of general medical outpatients attending the Hospital Clinic in Barcelona (N=385), and psychiatric diagnoses were made using DSM-IV criteria. A two-factor solution corresponding to the original two subscales of the HADS was found. The Spanish version of the HADS had good internal consistency and external validity, with favorable sensitivity and specificity in identifying cases of psychiatric disorder as defined by the Structured Clinical Interview for DSM-IV (SCID-I). The psychometric properties of the HADS and its brevity make it useful for screening for psychiatric disorders in the medically ill. PMID- 12850661 TI - Screening for autism spectrum disorder in adult psychiatric outpatients in a clinic in Taiwan. AB - Patients with adult autism spectrum disorder (ASD) continue to suffer from impairment in socialization and communication skills, and a proportion of them may develop psychiatric symptoms. It is thus likely that physicians in adult psychiatric departments may see a number of patients with ASD. Identification of patients with ASD is helpful and important for rehabilitation. This study estimated the prevalence of ASD among adult psychiatric outpatients in a Taiwanese medical center. A total of 660 patients were screened with Nylander and Gillberg's "Autism Spectrum Disorder in Adult Screening Questionnaire." Patients with high scores then underwent a diagnostic clinical interview conducted by child psychiatrists. Four patients (0.6%) were found to have ASD. PMID- 12850662 TI - Treatment for delirium with risperidone: results of a prospective open trial with 10 patients. AB - Delirium is a common psychiatric illness among medically compromised patients. There is an increasing opportunity to use atypical antipsychotics to treat delirium. The effects of these drugs on delirium, however, the most appropriate way to use them, and the associated adverse effects remain unclear. To clarify these points, a prospective open trial on risperidone was carried out in 10 patients with delirium. At a low dose of 1.7 mg/d, on average, risperidone was effective in 80% of patients, and the effect appeared within a few days. There were no serious adverse effects. However, sleepiness (30%) and mild drug-induced parkinsonism (10%) were observed; the symptom of sleepiness was a reason for not increasing the dose. One patient responded to a dose as low as 0.5 mg/d, so it is recommended that treatment start at a low dose, which may then be increased gradually. This trial is a preliminary open study with a small sample size, and further controlled studies will be necessary. PMID- 12850663 TI - Is pica in the spectrum of obsessive-compulsive disorders? PMID- 12850664 TI - The diagnosis of Asperger's syndrome in an adult presenting with an index episode of mania. PMID- 12850665 TI - Delirium associated with amitriptyline/fluconazole drug. PMID- 12850666 TI - Association of abnormal diurnal blood pressure variation with the development of silent cerebral infarction in patients with late-life-onset depression. PMID- 12850669 TI - Mechanism of interferon alpha on inhibition of metastasis and angiogenesis of hepatocellular carcinoma after curative resection in nude mice. AB - The aim of this study was to examine the mechanism of interferon alpha (IFN alpha) on inhibition of metastasis and recurrence of hepatocellular carcinoma (HCC). Nude mice bearing human HCC xenografts with high metastatic potential (LCI D20) underwent curative resection of tumors on postimplant day 11. IFN-alpha was begun the next day at different dosages given subcutaneously for 35 consecutive days; normal saline solution was injected into the control mice. The mice were killed 48 hours after the final treatment, and the parameters were evaluated. The HCC intrahepatic recurrence rate, the size of the recurrent lesions, the rate of lung metastasis, the serum vascular endothelial growth factor level, and the microvessel density (immunohistochemistry) were as follows: 100%, 2136+/-794 mm(3)(mean+/-standard deviation), 100%, 265.7+/-154.7 pg/ml, and 144+/-37/HP, respectively, in the control mice, whereas these same values were 62.5%, 89+/-45 mm(3), 12.5%, 53.3+/-9.9 pg/ml, and 86+/-25/HP, respectively, in the IFN-alpha 1.5 x 10(7)U/kg treatment group (P<0.05) and 26.7%, 46+/-21 mm(3), 0%, 65.2+/ 17.9 pg/ml, and 39+/-14/HP in the IFN-alpha 3 x 10(7)U/kg treatment group, respectively (P<0.05). However, a significant difference was not found in the serum levels of basic fibroblast growth factor among the control and IFN-alpha treatment groups. IFN-alpha inhibits metastasis and recurrence of human HCC after curative resection in nude mice mediated by antiangiogenesis through downregulating expression of vascular endothelial growth factor but not basic fibroblast growth factor. PMID- 12850670 TI - Achalasia and chest pain: effect of laparoscopic Heller myotomy. AB - Some patients with achalasia complain of chest pain in addition to dysphagia and regurgitation. Chest pain is said to be most common in young patients who have been symptomatic for a short time, and who often have vigorous achalasia (distal esophageal amplitude >/=37 mm Hg). Although pneumatic dilatation is reported to improve chest pain in 20% of patients, the effect of laparoscopic Heller myotomy on chest pain is unknown. The aim of this study was to determine the following in achalasia: (1) the prevalence of chest pain; (2) the clinical and manometric profiles of patients with chest pain; and (3) the effect of laparoscopic Heller myotomy. Between 1990 and 2001, a total of 211 patients with achalasia were studied (upper gastrointestinal series, esophagoduodenoscopy, and manometry). A total of 117 patients (55%) had chest pain in addition to dysphagia and regurgitation; 63 (54%) of these 117 patients underwent laparoscopic Heller myotomy and Dor fundoplication. Median follow up was 24 months. Age (49+/-16 years vs. 51+/-14 years [mean+/-SD]), duration of symptoms (71+/-91 months vs. 67+/-92 months [mean+/-SD]), and presence of vigorous achalasia (50% vs. 47%) were similar in those with and without chest pain. Ten (16%) of the 63 patients with chest pain who underwent Heller myotomy had vigorous achalasia. Postoperatively chest pain resolved in 84% and improved in 11% of patients. There was no difference in clinical outcome between patients with and without vigorous achalasia. These data demonstrate the following: (1) chest pain was present in 55% of patients with esophageal achalasia; (2) chest pain was not related to age, duration of symptoms, or manometric findings; and (3) laparoscopic Heller myotomy improved chest pain in 95% of patients, regardless of the manometric findings. Thus laparoscopic Heller myotomy was highly effective in treating achalasia with chest pain. PMID- 12850671 TI - Intratracheal long-term pH monitoring: a new method to evaluate episodes of silent acid aspiration in patients after esophagectomy and gastric pull up. AB - Aspiration of gastric contents is considered a leading cause of postoperative pulmonary complications after esophagectomy and gastric pull up but has been difficult to diagnose. We used intratracheal long-term pH monitoring to evaluate the prevalence of aspiration of gastric contents in patients undergoing these operations. Continuous intratracheal pH monitoring was carried out during the first 72 postoperative hours in 16 patients with esophageal carcinoma who had undergone esophagectomy and gastric pull up. A drop in the pH to less than 4 was defined as an episode of acid aspiration. All patients except one tolerated the probe without any difficulties. Episodes of acid aspiration could be detected in 12 (80%) of 15 patients (5 of 8 after transhiatal esophagectomy, 7 of 7 after transthoracic esophagectomy, 2 of 5 with reconstruction in the anterior mediastinum, and 9 of 10 with reconstruction in the posterior mediastinum). The number of aspiration episodes was significantly higher during postoperative day 1 (P=0.03) compared to postoperative days 2 and 3. Two patients developed pneumonia later in the postoperative course. Both of them had several episodes of acid aspiration detected by pH monitoring immediately postoperatively. Intratracheal pH monitoring is a safe, feasible, and well-tolerated method for detecting episodes of acid aspiration after esophagectomy and gastric pull up. Aspiration of gastric contents is a common phenomenon particularly during the first 24 postoperative hours after transthoracic esophagectomy and gastric pull up in the posterior mediastinum and appears to correlate with the development of postoperative pneumonia. PMID- 12850672 TI - Behavior of plasma hemoglobin in an experimental model of occlusive mesenteric ischemia. AB - The difficulties in establishing the diagnosis of mesenteric ischemia are responsible for the high mortality rate that is associated with this clinical condition. We studied the behavior of plasma hemoglobin in an experimental model of occlusive mesenteric ischemia in mice. Our results showed a clear relationship between the duration of ischemia and plasma hemoglobin levels. With regard to the time frames studied (3 hours, 6 hours, 12 hours, and 24 hours), comparison with control groups produced calculated P values of less than 0.01 for all time frames with the exception of the 3-hour group. This test may have the potential to aid in the diagnosis of mesenteric ischemia as well as the follow-up of its course after various therapeutic approaches. PMID- 12850674 TI - Solitary necrotic nodule of the liver: a riddle that is difficult to answer. AB - Solitary necrotic nodule of the liver is an unusual lesion that is often an incidental finding on abdominal imaging, intraoperative examination, or post mortem. Most reported cases of solitary necrotic nodule have been in males, and over three quarters of these lesions have occurred in the right lobe of the liver. Pathologically, solitary necrotic nodule is a benign lesion characterized by a completely necrotic core that is often partly calcified, surrounded by a dense hyalinized fibrous capsule containing elastin fibres. The ultrasound appearance of solitary necrotic nodule is usually of a "target" lesion with a hyperechoic center, while on CT scan they appear as non-enhancing hypodense lesions that are typical of metastatic adenocarcinoma or peripheral cholangiocarcinoma. The impression of malignancy is further enforced with the finding of necrotic cellular material on biopsy and the macroscopically hard and "gritty" nature of the nodules. Currently, permanent histopathology of solitary necrotic nodules is the only accurate method of diagnosis. However, solitary necrotic nodules are usually of a bilobed or lobulated shape that is unusual for malignant liver lesions, and they often lie in close proximity to hepatic inflow structures. Solitary necrotic nodule should be suspected in liver lesions with this configuration, location, and on a biopsy showing a large amount of necrosis. PMID- 12850673 TI - Evaluation and management of patients with recurrent peptic ulcer disease after acid-reducing operations: a systematic review. AB - This systematic review examines the evidence for commonly employed strategies of managing patients with recurrent ulcer disease after acid-reducing operations. Particular attention is given to recent evidence relating Helicobacter pylori (H. pylori ) and nonsteroidal anti-inflammatory drugs (NSAIDs) to ulcer recurrence after operative therapy. MEDLINE word searches of the literature from 1966 to 2001 identified 895 articles that cross-reference the terms "peptic ulcer disease (PUD)," "surgery," and "recurrence." Articles were selected for systematic review of evidence relating incomplete vagotomy, NSAIDs, and H. pylori to postoperative ulcer recurrence and evidence supporting common medical and surgical strategies. The relationship between incomplete vagotomy and recurrent ulcer disease is suggested by randomized controlled trials and well-designed prospective case series. The evidence that NSAID use is an important pathogenic factor in recurrent ulcer disease includes the relationship between NSAIDs and primary PUD, the occurrence of NSAID-induced ulcers in patients taking proton pump inhibitors, and case series demonstrating virulent ulcer disease in patients taking aspirin despite prior acid-reducing operations. The relationship between H. pylori infection and postoperative ulcer recurrence remains uncertain despite multiple controlled trials and well-designed case series that have documented high rates of H. pylori infection in postoperative patients. The initial management of patients with recurrent ulcer disease after acid-reducing operations consists of a protein pump inhibitor or a histamine-2 receptor antagonist and antibiotics directed at H. pylori, if present. Evidence for this regimen includes prospective randomized trials demonstrating the efficacy of cimetidine in healing ulcers after acid-reducing operations and prospective, randomized studies documenting the efficacy of histamine-2 receptor antagonists and protein pump inhibitors in the management of patients with primary PUD. The critical role that H. pylori infection plays in primary PUD and the minimal risks associated with H. pylori eradication strongly support the initiation of antibiotic therapy when H. pylori is present. The principal indication for operative management of recurrent PUD is the occurrence of ulcer complications that cannot be managed by medical or endoscopic means. The operative management of patients with failed acid-reducing operations is based on ulcer recurrence rates and morbidity and mortality rates in randomized and nonrandomized prospective trials of patients with primary PUD and retrospective case series of patients undergoing remedial operative procedures after various failed acid-reducing operations. PMID- 12850675 TI - Small cell carcinoma of the cystic duct: a case report. AB - Small cell carcinoma usually involves the lung and rarely affects the biliary tract, especially the cystic duct. In this article we report a case of small cell carcinoma of the cystic duct in a 46-year-old Japanese man. The patient presented with abdominal pain and jaundice. Imaging showed a small nodule in the cystic duct invading the common bile duct with dilatation of the proximal biliary tree. The hepatic artery and portal vein were free from invasion. Extended right hepatic lobectomy, cholecystectomy, and resection of the extrahepatic proximal bile ducts were performed together with lymph node dissection under the tentative diagnosis of carcinoma of the cystic duct. Histopathologic examination of the resected specimen revealed small cell carcinoma arising in the cystic duct and extending into the common bile duct. The postoperative clinical course was uneventful, and the patient is doing well without any signs of recurrence 1 year after the operation. To our knowledge this is the first documented case of a small cell carcinoma arising in the cystic duct. PMID- 12850676 TI - Spontaneous and traumatic intra-peritoneal perforations of hepatic hydatid cysts: a case series. AB - Little is known about the presentation, management, outcome, and recurrence of hydatid cyst perforations. We reviewed the charts of all patients admitted to our emergency service for 7 years to identify patients who were surgically treated for intra-peritoneal hydatid cyst perforations. Twelve hydatid cysts were identified in 7 patients (5 males; median age 22 yr; range 8-67). The perforations occurred spontaneously in 5 patients, and were the result of mild trauma in 2 patients. Diagnostic tools included ultrasound (US, n = 4), computed tomography (CT, n = 3), and diagnostic peritoneal lavage (DPL, n = 1). The cysts were treated with radical (n = 3) or conservative (n = 9) operative techniques. Intra-cavitary and intra-abdominal spaces were washed in 6 and 5 patients, respectively. The median follow-up time was 41 months (range 3-58). Indirect hemagglutination test was positive in 3 patients, but CT confirmed cyst recurrence in only 2 of these patients. Both had had large cysts and had undergone conservative therapy (endocystectomy and external drainage). An intra abdominal recurrence was observed in a patient whose abdomen had not been washed during surgery. In conclusion, patients with hydatid cyst perforations in our study generally presented with severe abdominal findings. US, CT, and DPL may be helpful for the diagnosis. Recurrence may be related to operative technique, location of the cyst, and abdominal wash during the surgery. PMID- 12850677 TI - Management of acute cholecystitis in the laparoscopic era: results of a prospective, randomized clinical trial. AB - The aim of this prospective, randomized study was to determine whether laparoscopic cholecystectomy should be performed as an early or a delayed operation in patients with acute cholecystitis. After diagnostic workup, patients were randomized to one of two groups: (1) early laparoscopic cholecystectomy (i.e., within 7 days after onset of symptoms) or (2) initial conservative treatment followed by delayed laparoscopic cholecystectomy 6 to 8 weeks later. Seventy-four patients were placed in the early-operation group, and 71 patients were assigned to the delayed-operation strategy. There was no significant difference in conversion rates (early 31% vs. delayed 29%), operating times (early 98 [range 30 to 355] minutes vs. delayed 100 [45 to 280] minutes), or complications. Failure with the conservative treatment strategy was noted in 26% of these patients. The total hospital stay was significantly shorter in the early group (5 [range 3 to 63] days) vs. the delayed group (8 [range 4 to 50] days; P<0.05). Despite a high conversion rate, early laparoscopic cholecystectomy offered significant advantages in the management of acute cholecystitis compared to a conservative strategy. The greatest advantage was a reduced total hospital stay. PMID- 12850678 TI - Staged abdominal repair in the treatment of intra-abdominal infection: analysis of 102 patients. AB - Surgical treatment of intra-abdominal infections remains a challenge for the surgeon. Staged abdominal repair is being commonly used in patients with intra abdominal infections. This study presents our experience with staged abdominal repair and analyzes factors affecting mortality. A total of 102 patients who underwent staged abdominal repair procedures for intra-abdominal infections during a 12-year period were retrospectively reviewed. The effects of several risk factors on mortality were evaluated. The investigated risk factors included age, Acute Physiology and Chronic Health Evaluation II (APACHE II) score, number of operations prior to staged abdominal repair, number of repeat laparotomies, anatomic origin of infection, and etiology of intra-abdominal infections. The overall mortality rate was 40% (41/102). The mean number of operations prior to staged abdominal repair (0.72 +/- 0.1 in survivors vs. 1.37 +/- 0.21 in nonsurvivors), age (24.5% mortality under 55 years vs. 53.6% mortality between 55 and 65 years vs. 75% mortality over 65 years), and APACHE II score (13.4 +/- 3.4 in survivors vs. 20.3 +/- 6.64 in nonsurvivors) were correlated with mortality rates (P < 0.05). Our results showed that the physiologic status of patients, severity of sepsis, and decision time for staged abdominal repair were all associated with higher mortality. PMID- 12850679 TI - Watermelon stomach: pathophysiology, diagnosis, and management. AB - Watermelon Stomach (WS) has been increasingly recognized as an important cause of occult gastrointestinal blood loss. Clinically, patients develop significant iron deficiency anemia and are frequently transfusion dependent. The histologic hallmark of WS is superficial fibromuscular hyperplasia of gastric antral mucosa with capillary ectasia and microvascular thrombosis in the lamina propria. Endoscopic findings of the longitudinal antral folds containing visible columns of tortuous red ectatic vessels (watermelon stripes) are pathognomonic for WS. Trauma to the mucosal epithelium overlying engorged vessels by gastric acid or intraluminal food results in bleeding. Treatment options for WS include endoscopic, pharmacologic, and surgical approaches. Endoscopic therapy, including contact and non-contact thermal ablations of the angiodysplastic lesions, is the mainstay of conservative therapy. However, many patients fail endoscopic therapy and develop recurrent acute and chronic GI bleeding episodes. Surgical resection may be the only reliable method for achieving a cure and eliminating transfusion dependency. Traditionally, surgery was used only as a last resort after patients failed prolonged medical and/or endoscopic therapy. However, based on the experience garnered from the literature we recommend a more aggressive surgical approach in patients who fail a short trial of endoluminal therapy. PMID- 12850680 TI - Morphology and function of canine small intestinal autografts: with particular interest in the influence of ex vivo graft irradiation. AB - The intestinal transplantation procedure obligates an early functional deficit in intestinal grafts. Graft irradiation has been used to modulate post-transplant immune reactions; however, irradiation may cause further deterioration of the function of the transplanted intestine. Using the canine model, we investigated the influence of the transplant procedures, with and without ex vivo graft irradiation, on early intestinal graft function and histopathology. Outbred hound dogs underwent autointestinal transplantation with (n=8) or without (n=5) 7.5 Gy ex vivo graft irradiation. Mucosal cytochrome P450 and P-glycoprotein, routine immunohistopathology, and intestinal absorptive function were studied. Weight gain was slow after surgery, but was comparable in the irradiated and nonirradiated groups. During the early post-transplant period, both groups showed defects in intestinal absorption, associated with decreased cytochrome P450 3A4 activity and reduced P-glycoprotein expression, regardless of graft irradiation. These changes returned to normal in both groups by day 28. Histopathologically, epithelial apoptosis showed a slight increase 1 hour after transplantation; however, there was no evidence of histopathologic abnormalities including arterial changes associated with irradiation. In addition, the frequency of T and B lymphocytes in the lamina propria were not significantly influenced by the transplant surgery or ex vivo irradiation. Thus, early after transplantation, intestinal function was impaired and effectiveness of orally administered immunosuppressive drugs was significantly altered. Graft irradiation did not induce further defects in intestinal function or cause histopathologic abnormalities. PMID- 12850681 TI - A comparison of pancreaticogastrostomy and pancreaticojejunostomy following pancreaticoduodenectomy. AB - This retrospective study compares the results of pancreaticogastrostomy (PG) and pancreaticojejunostomy (PJ) in our institution, which has extensive experience in both techniques. Between the years of June 1995 and June 2001, 214 patients underwent pancreaticoduodenectomy (PD) at our institution. Of these 177 had PG and 97 had pancreatojejunostomy (PJ). There were 117 (54.6%) males and 97 (45.3%) females with a mean age of 64.2 +/- 12.4 years. Indications for surgery were pancreatic adenocarcinoma in 101 (47.2%), ampullary adenocarcinoma in 36 (16.9%), distal bile duct adenocarcinoma in 22 (10.2%), duodenal adenocarcinoma in 9 (4.2%), and miscellaneous causes in 46 (21.4%) of patients. Preoperatively, significant differences in the groups were that the patients undergoing PJ were significantly younger than those undergoing PG. Also noted preoperatively, was that the patients undergoing PG had a significantly lower direct bilirubin than those undergoing PJ. With regard to intraoperative parameters, operative time was significantly shorter in the PJ group when compared to the PG group. When the patients who did not develop fistula (N = 186) were compared to those who developed fistula (N = 28) the significant differences were that the patients who developed fistula were more likely to have hypertension preoperatively and a higher alkaline phosphatase. They also showed a significantly higher drain amylase and were likely to have surgery for ampullary, distal bile duct or duodenal carcinoma rather than pancreatic adenocarcinoma. In addition, those patients who developed fistula had a significantly longer postoperative stay, a larger number of intraabdominal abscesses and leaks at the biliary anastomosis. Thirty-day mortality was significantly higher in the PJ group compared to the PG (4 vs. 0, P = 0.041). There was a significantly larger number of bile leaks in the PJ group when compared to the PG (6 vs. 1, P = 0.048). In addition, the PJ group required a significantly larger number of new CT guided drains to control infection (8 vs. 2, P = 0.046) and the PJ group required a larger number of re explorations to control infection or bleeding (5 vs. 0, P = 0.018). However, the pancreatic fistula rate was not different between the two groups (12% [PG] vs. 14% [PJ]). This retrospective analysis shows that safety of PG can be performed safely and is associated with less complications than PJ and proposes PG as a suitable and safe alternative to PJ for the management of the pancreatic remnant following PD. PMID- 12850682 TI - Intra-abdominal hemorrhage due to rupture of a splenic vein aneurysm: a case report. AB - Splenic vein aneurysm (SVA) is extremely rare. Most patients with an SVA have portal hypertension. In this report we describe the first recorded case of intra abdominal hemorrhage due to rupture of an SVA in a patient without evidence of portal hypertension. A 72-year-old man was admitted to our medical center in a state of shock, with complaints of acute abdominal pain and abdominal distention. Preoperatively, abdominal ultrasonography demonstrated an echo-free space in the abdomen, suggesting the presence of a fluid collection. In addition, computed tomography revealed an enhanced lesion with contrast material in the pancreatic tail. An emergency operation showed bleeding from the SVA near the pancreatic tail. Consequently, a distal pancreatectomy with splenectomy was performed. Histologically the lesion was diagnosed as an SVA surrounded by pancreatic tissue with chronic inflammatory changes. The patient's postoperative course was uneventful. PMID- 12850683 TI - Intraoperative imprint cytology for evaluation of sentinel lymph nodes from visceral malignancies. AB - Although originally described for breast cancer and melanoma, sentinel lymph node (SLN) mapping techniques are being investigated in the treatment of visceral malignancies. There is no literature evaluating intraoperative analysis of SLNs from visceral sites. We evaluated the utility of touch preparation intraoperative imprint cytology (IIC) in evaluating SLNs harvested in the setting of visceral malignancy. SLN mapping procedures involving 50 cases of visceral malignancy (37 colon, 12 gastric, and 1 small bowel), from February 1999 through August 2001, were studied. In each case, subserosal injections of isosulfan blue were used to identify the SLN. The SLNs were then sent fresh to the pathology laboratory for evaluation by IIC. A standard lymphadenectomy was performed in all cases. Postoperatively, the SLNs were evaluated by means of using hematoxylin and eosin staining. If these stains were normal, immunohistochemical analyses using carcinoembryonic antigen and cytokeratin were subsequently performed. SLNs were successfully identified in 46 cases (92%), and a total of 95 SLNs were harvested. The average number of SLNs was 1.9 with a range of one to six. More SLNs were found with gastric than with colonic lesions (2.8 vs. 1.8; P=.017). Evaluable IIC in 41 cases revealed metastatic disease in 10 SLNs, representing seven patients. Of the 34 patients with normal IIC, five were found to have positive SLNs on hematoxylin and eosin staining. An additional three patients were found to have positive SLNs only on immunohistochemical analysis. The overall sensitivity and specificity of IIC was 64% and 100%, respectively. This resulted in a positive predictive value of 100% and a negative predictive value of 86%. The use of IIC to evaluate SLNs from visceral malignancies is clearly feasible. When the IIC of the SLN is positive, the surgeon may feel confident that disease is actually present in the SLN. If there is a negative result, the technique may miss disease that is present on subsequent permanent sections. We do not recommend routine use of IIC; however, it may be of use in clinical trials. PMID- 12850684 TI - The hypertensive lower esophageal sphincter: a motility disorder with manometric features of outflow obstruction. AB - The aim of this study was to define the clinical presentation, motility characteristics, and prevalence and patterns of gastroesophageal reflux in patients with hypertensive lower esophageal sphincter (HTLES). HTLES was defined by a resting pressure measured at the respiratory inversion point on stationary manometry of greater than 26 mm Hg (ninety-fifth percentile of normal). One hundred consecutive patients (80 women, 20 men; mean age 54.7 years, range 23 to 89 years), diagnosed with HTLES at our institution between September 1996 and October 1999, were studied. Patients with achalasia or other named esophageal motility disorders or history of foregut surgery were excluded, but patients with both HTLES and "nutcracker esophagus" were included. The most common symptoms in patients with HTLES were regurgitation (75%), heartburn (71%), dysphagia (71%), and chest pain (49%). The most common primary presenting symptoms were heartburn and dysphagia. The intrabolus pressure, which is a manometric measure of outflow obstruction, was significantly higher in patients with HTLES compared to normal volunteers. The residual pressure measured during LES relaxation induced by a water swallow was also significantly higher than in normal persons. There were no significant associations between any of the relaxation parameters studied (residual pressure, nadir pressure, duration of relaxation, time to residual pressure) and either the presence or severity of any symptoms or the presence of abnormal esophageal acid exposure. Seventy-three patients underwent 24-hour pH monitoring, and 26% had increased distal esophageal acid exposure. Compared to a cohort of patients with gastroesophageal reflux disease but no HTLES (n=300), the total and supine periods of distal esophageal acid exposure were significantly lower in the patients with HTLES and abnormal acid exposure. Patients with HTLES frequently present with moderately severe dysphagia and typical reflux symptoms. Approximately one quarter of them have abnormal esophageal acid exposure on pH monitoring. Patients with HTLES have significantly elevated intrabolus and residual relaxation pressures on liquid boluses, suggesting that outflow obstruction is present. PMID- 12850685 TI - Effect of colonic distention on ileal motor activity with evidence of coloileal reflex. AB - Chyme delivery from the ileum to the colon is controlled by various neurologic and hormonal factors, many of which remain to be identified. In this report we investigated the effect of colonic distention on ileal motility with the aim of identifying the mechanism of chyme delivery from the ileum to the colon. The right colon of 16 healthy volunteers (12 men and 4 women; mean age 36+/-9 years standard deviation) was distended by a balloon that was filled with saline solution in increments of 20 ml. The pressure response of the terminal ileum to the colonic distention was recorded by a saline-perfused tube. The test was repeated in nine subjects after the colonic segment around the balloon was anesthetized by xylocaine injection into the colonic wall. Twenty and 40 ml colonic distention produced no significant ileal pressure response. Colonic distention with 60 ml produced an increase in colonic pressure (P<0.05), as measured by intraballoon pressure, and a decrease in ileal pressure (P<0.05); a similar response was achieved with 80 ml distention. At 100 ml colonic distention, the balloon was dispelled to the transverse colon. Distention up to 100 ml of the anesthetized colonic segment produced no significant colonic or ileal pressure response. The flow of chyme from the small to the large gut appears to be controlled by a reflex mechanism that we call the "coloileal reflex." Whenever the right colon is distended with a substantial volume of chyme that increases the intraluminal pressure, it is suggested that ileal relaxation occurs, which delays the emptying of chyme from the ileum. PMID- 12850686 TI - Patients with Crohn's disease are unaware of the risks that smoking has on their disease. AB - Smoking, both in quantity and duration, increases the risk of recurrent Crohn's disease (CD) and the need for surgical treatment of CD. This study assessed awareness among patients with CD of the risks of smoking on their disease. We distributed self-administered questionnaires to 714 CD patients under follow-up at the University Hospital Birmingham, United Kingdom. We asked the patients eight multiple-choice (yes/uncertain/no) questions about the effects of smoking on health and on CD. We also determined relevant patient demographics, smoking history, and medical history. A total of 312 patients completed the questionnaire. Patients acknowledged the dangers of smoking on overall health (91.5%), lung cancer (89.6%), lung disease (90.8%), and cardiovascular disease (85.3%). Employed, educated, and homeowning patients demonstrated significantly more recognition of these smoking risks. Conversely, few patients recognized that smoking increases the risks of development of CD (9.5%) and of reoperation for CD (12.0%). Moreover, few patients recognized that both the quantity of cigarettes smoked (11.4%) and the duration of smoking (17.4%) increase the risk of reoperation. Patients with CD are unaware of the risks that smoking has on their disease, indicating a need for increased patient education with regard to the effects of smoking on CD. PMID- 12850687 TI - A randomised, open label, controlled trial on the effect of interferon plus amantadine compared with interferon alone for treatment of chronic hepatitis C. AB - BACKGROUND: Combination of the antiviral drug amantadine with interferon (IFN) may be more effective than IFN monotherapy for treatment of chronic hepatitis C. METHODS: We randomised 93 patients with chronic hepatitis C to IFNIFN 6 MU 3 times/week for 24 weeks, followed by IFN 3 MU 3 times/week for further 24 weeks given in combination with amantadine 100 mg t.i.d. (regimen A, n=48) or as monotherapy (regimen B, n=45). Control liver biopsies were obtained 6 months post treatment. RESULTS: At the end of the trial a similar proportion of patients had normal serum ALT levels (35% for regimen A, and 44% for regimen B) as measured by intention to treat criteria. Sustained biochemical response at 6 months post treatment was 15 and 20%, and sustained virological response was 10 and 20% for regimen A and B, respectively. The effect on liver histology was also similar: the inflammatory components of the Knodell score decreased by 1.3+/-0.6 points for regimen A and by 1.6+/-0.6 for regimen B, and score for fibrosis remained unchanged with both regimens. CONCLUSIONS: Combination of IFN therapy with amantadine does not enhance the effect of IFN as shown by biochemical, virological and histological criteria. PMID- 12850688 TI - Endoscopic color Doppler ultrasonographic evaluation of recurrent esophagogastric varices following endoscopic injection sclerotherapy. AB - BACKGROUND: Endoscopic injection sclerotherapy (EIS) for treatment of esophagogastric varices is well established in Japan. However, varices may still recur unpredictably following EIS. We studied this problem using endoscopic color Doppler ultrasonography (ECDUS) and specifically examined esophagogastric blood flows. METHODS: Prophylactic EIS was performed by intravariceal injection of 5% ethanolamine oleate (EO) in 49 patients with esophageal varices secondary to liver cirrhosis. No patient had documented hepatocellular carcinoma (HCC) before EIS, and patients who developed HCC during follow-up were excluded. We performed ECDUS before EIS, and 2 weeks and 2 years later. The esophagogastric intra- and extramural venous blood flows, including flow in the azygos vein, were compared between these observations. Gastric intramural blood flow and changes in extramural gastric blood flow, including the azygos vein flow, were scored. Dynamic computer tomography (CT), ultrasonography (US), and color Doppler ultrasonography (CDUS) were also performed before EIS and 1 month following the procedure. Thereafter, patients underwent CT and US examinations every 6 months for 2 years to detect any development of porto-systemic shunts or HCC. RESULTS: The average number of EIS procedure per patient was 3.1+/-0.8 (mean+/-SD), and the total amount of sclerosant injected was approximately 33.5+/-6.5 ml. The overall recurrence rate over the 2-year follow-up was 36.7%. The gastric intra- and extramural blood flows did not differ between those patients with or without major shunts before EIS. In patients with recurrent variceal formation, the gastric intramural blood flow score following EIS (2.1+/-0.5) was significantly higher than that in patients without recurrence (0.8+/-0.6) (P<0.01). In addition, gastric extramural blood flow score following EIS (0.8+/-0.6) was significantly lower in patients with recurrence than that in those without recurrence (1.7+/-0.5) (P<0.01). The same differences held after exclusion of patients with major shunts. The gastric intramural blood flow score in patients with recurrent variceal formation (2.1+/-0.4) was significantly higher than that in patients without recurrence score (P<0.01). Moreover, gastric extramural blood flow score in patients with recurrent variceal formation (1.0+/-0.7) was significantly lower than in patients without recurrence (1.6+/-0.5) (P<0.01). CONCLUSIONS: Two characteristics were observed in patients with recurrent cases of esophageal varices 2 weeks following EIS. The first was the maintenance of gastric intramural blood flow. The second was the absence of dilation of the gastric extramural blood vessels. These observations may enable us to predict the recurrence of esophagogastric varices using ECDUS within 2 weeks following EIS. PMID- 12850689 TI - Cardiac dysfunction in cirrhosis. AB - In cirrhosis, cardiac symptoms and physical signs occur as the liver functions worsen. Cirrhosis is associated with hyperdynamic circulation and beta-adrenergic system activation responsible for the cardiovascular changes. The purpose of the present study was to explore the cardiovascular response to exercise in cirrhotic patients. A total of 20 patients (16 men, four women) with cirrhosis of hepatitis B or C without any cardiac dysfunction were included in the study. Ten people (eight men, two women) were enrolled in the control group. Plasma noradrenaline and adrenaline concentrations, blood pressures in supine and standing positions, exercise echocardiography and exercise radionuclide ventriculography were carried out. In cirrhotics, the exercise capacity was lower and also there was an impaired cardiovascular response to exercise with lower than expected peak heart rate without any cardiac dysfunction, which may have important clinical implications for the ability of these patients to withstand cardiovascular stress. PMID- 12850690 TI - Expression of DNA (5-cytosin)-methyltransferases (DNMTs) in hepatocellular carcinomas. AB - Down-regulation of tumor suppressor genes by hypermethylation of 5'-CpGs is one of the important mechanisms involved in tumor development. DNA (5-cytosine) methyltransferases (DNMTs) are enzymes that methylate the cytosine residue of CpGs, and four types have been identified (DNMT1, 2, 3a and 3b). To examine the involvement of DNMTs in hepatocellular carcinogenesis, we measured DNMT mRNAs in hepatocellular carcinomas (HCCs). mRNAs of DNMT1, 2, 3a and 3b were detected by reverse transcription-PCR analysis and quantified by a real-time PCR method in surgically resected HCCs and adjacent non-tumorous liver tissue. DNMT1 was expressed in all tissues and at a significantly higher level in HCCs than in non tumorous liver tissue (P=0.01). DNMT2 was expressed at a low level in all tissues. DNMT3a and DNMT3b mRNA were undetectable in normal liver. DNMT3a was expressed in all HCCs and was expressed at similar levels in 60% of the non tumorous liver tissues. DNMT3b mRNA was detected at a significantly higher level (P=0.002) in HCCs than in non-tumorous liver tissues. The amount of DNMT1, 3a and 3b mRNA was not different between HCCs with or without hypermethylation of the CDH1 promoter. These data suggest that overexpression of DNMT1 and DNMT3b contributes to hepatocellular carcinogenesis. PMID- 12850691 TI - Heparanase expression in hepatocellular carcinoma and the cirrhotic liver. AB - The endoglycosidase heparanase plays an important role in tumor cell invasion, angiogenesis and metastasis. In this study, we analyzed heparanase expression in primary hepatocellular carcinoma (HCC), liver cirrhosis and normal liver tissues to further evaluate its role in tumor pathogenesis. Heparanase mRNA expression was measured by real-time quantitative RT-PCR and localized at the cellular level by in situ hybridization. Heparanase protein levels and its localization were determined by Western blotting and immunohistochemistry. Expression of heparanase mRNA in primary HCC was increased 2-fold compared with liver cirrhosis and 2.2 fold in comparison with normal liver tissues, and this overexpression was even more pronounced in advanced stage HCC. In contrast, heparanase expression levels between cirrhotic tissues and normal liver tissues were not significantly different. In HCC the increased heparanase expression was localized in hepatic tumor cells and was only weakly present or absent in normal hepatocytes, bile duct epithelial cells and the connective tissues. These results suggest that increased heparanase expression is involved in the pathogenesis and progression of HCC. Its specific up-regulation in HCC but not in liver cirrhosis indicates that it might be used as a molecular marker for the differentiation of these disorders. PMID- 12850692 TI - DNA alterations during multi-step development of human hepatocellular carcinomas revealed by laser capture microdissection. AB - In order to clarify early molecular events involved in liver carcinogenesis, we analyzed 53 liver-cirrhosis nodules (LCNs) from five patients and 13 micro hepatocellular carcinoma (HCC) nodules from one patient and looked for alterations of microsatellites in genomic DNA after carefully preparing the tissue samples by laser-capture microdissection (LCM). Allelotyping was done with 20 markers corresponding to anonymous microsatellites and 13 corresponding to tumor suppressor genes (TSGs) that had shown significant alterations in HCCs. We detected both loss of heterozygosity (LOH) and microsatellite shifts (MS). Overall, 24 of 53 (47%) of LCNs showed LOH with any of the informative markers used in the study, reflecting that proportion of LCNs with clonal growth. The fractional allelic loss (FAL) index, an indication of total genomic complexity, was not significantly different between LCN and micro-HCC nodules, but their profiles of alteration were different. These profiles were classified into three groups: (1) LCN profile-allelic loss at chromosomal arms 1q and 14q, TBP and BRCA1; (2) HCC profile-LOH at 4q, 6q, 7q, 17p, NF1, IGFIIr and p53 in micro-HCC nodules; these changes in early lesions were identical to those seen in mature HCCs; (3) Common profile-LOH at NF1 and 6q, including IGFIIr, common to both LCN and HCC. No LCN showed LOH at p53 and Rb, loci that are generally altered in HCCs. However, 12 intra-tumoral nodules examined had lost p53 in all informative cases, although the loss of Rb was a late event. These results suggest that early genomic profiles confined to LCNs, and additional profiles that can be observed when liver tissue undergoes malignant transformation, support a model of multi step development of HCC. PMID- 12850693 TI - Activation of hepatic macrophage contributes to hepatic necrosis after post ischemic reperfusion in alcoholic fatty liver. AB - Fatty livers are vulnerable to ischemia/reperfusion (I/R) injury. We investigated the role of hepatic macrophages in the I/R injury in the fatty liver. Rats with alcoholic or nonalcoholic fatty liver were subjected to hepatic warm ischemia for 30 min. A bolus of gadolinium chloride (GdCl(3)) was injected intravenously twice before I/R to block hepatic macrophage activity. Alcoholic fatty liver developed more extensive hepatic necrosis with neutrophil infiltration in association with a higher production of cytokine-induced neutrophil chemoattractant (CINC)-1, a potent neutrophil chemokine in rat, after I/R than the nonalcoholic fatty liver or control liver without steatosis. Hepatic apoptosis after I/R increased to a similar degree (3-fold) in each of the two fatty liver models, compared with the control liver. Alcoholic fatty liver exposed to I/R showed a rapid increase in nuclear factor-kappa B (NF-kappaB) binding activity. The GdCl(3) pretreatment significantly reduced NF-kappaB binding activity, CINC-1 level and necrosis in alcoholic fatty liver, despite no significant decrease in the extent of apoptosis. Our results suggest that the activation of hepatic macrophages in alcoholic fatty liver may contribute to hepatic necrosis after I/R, and that the apoptosis might be less dependent on the macrophage activity. PMID- 12850694 TI - Protection against post-ischemic mitochondrial injury in rat liver by silymarin or TUDC. AB - BACKGROUND: Hepatic ischemia/reperfusion (I/R) injury is characterized by features such as cholestasis. Mitochondria become susceptible to damage during ischemia and after reperfusion. The aim of this study was to determine if silymarin and tauroursodeoxycholic acid (TUDC), would prevent impairment of liver mitochondrial function following I/R injury. METHODS: Livers of male Wistar rats were subjected to 45 min of hepatic ischemia. During the 90 min of reperfusion, livers were perfused with either vehicle, silymarin, or TUDC. Changes in membrane potential, mitochondrial respiration as well as susceptibility to mitochondrial permeability transition (MPT) induction were evaluated and endogenous adenine nucleotides were measured. RESULTS: In rats subjected to I/R, compared with the control group, a severe impairment of mitochondrial bioenergetics was observed. State 3 respiration was decreased and state 4 enhanced, associated with lower membrane potential developed following succinate energization. An increased susceptibility to MPT induction by calcium/phosphate was also observed. The effects of I/R injury were ameliorated in the presence of silymarin but not TUDC. Similarly, ATP levels following I/R were lower, in comparison with the control group and the silymarin treatment but not TUDC. CONCLUSION: Thus, silymarin, but not TUDC, prevents the most significant changes that occur in mitochondria during I/R, and probably, the associated cell dysfunction, through their central role in cellular bioenergetics. PMID- 12850695 TI - In vitro differentiation of hepatocyte-like cells from embryonic stem cells promoted by gene transfer of hepatocyte nuclear factor 3 beta. AB - We investigated the ability of a genetically altered embryonic stem (ES) cell line to promote endodermal differentiation toward hepatocytes in vitro by transfecting the hepatocyte nuclear factor-3beta (HNF-3beta) gene. Parental and HNF-3beta-transfected ES cells were initiated toward differentiation in embryoid bodies (EBs) for 5 days and the resulting EBs were transferred to an attached culture system. Albumin production was observed using an immuno-cytochemical method 7 days after induction of differentiation in almost all differentiating HNF-3beta-transfected ES cells, whereas scant immuno-reactivity against albumin was found on the same day in the cultures of differentiating parental ES cells. An analysis using a reverse transcriptase polymerase chain reaction revealed the HNF-4alpha expression in the HNF-3beta-transfected ES cells and also demonstrated that the expression of endodermal and hepatocyte-related markers, such as transthyretin, alpha-fetoprotein, albumin, alpha-1 antitrypsin, tryptophan-2,3 dioxygenase and phosphoenol-pyruvate carboxykinase, could be observed at an early stage in the outgrowths of HNF-3beta-transfected ES cells compared to the parental ES cells. These results suggest that HNF-3beta-transfected ES cells may be useful for the efficient induction of hepatocytes in vivo. PMID- 12850696 TI - ERK/MAPK-dependent PI3K/Akt phosphorylation through VEGFR-1 after VEGF stimulation in activated hepatic stellate cells. AB - Vascular endothelial growth factor (VEGF) can induce proliferation of endothelial cells through VEGFR-1 and VEGFR-2 as its receptors, but the intracellular signaling pathway via VEGFR-1 is still unclear. Previously we reported that stellate cells expressed VEGFR-1 exclusively during activation in the liver. In the present paper, the signaling pathway via VEGFR-1 was studied using rat stellate cells activated in vitro. Western blot analysis revealed that both ERK/mitogen-activated protein kinase (MAPK) and PI3K/Akt were phosphorylated in the cells activated by culture in Dulbecco's modified Eagle medium containing 10% fetal calf serum on plastic dishes for 9 days. When VEGF was added to the culture medium, the extent of such PI3K/Akt phosphorylation was increased despite that DNA synthesis and ERK/MAPK phosphorylation were unchanged in the cells. However, this up-regulation of PI3K/Akt phosphorylation was markedly diminished following addition of GFX and PD-98059, inhibitors for protein kinase C (PKC) and ERK/MAPK, respectively. Also, addition of GFX reduced phosphorylation of ERK/MAPK in the cells. It is suggested that VEGF may stimulate signal transduction of PI3K/Akt through VEGFR-1 dependently on activation of the PKC and ERK/MAPK pathway in activated hepatic stellate cells. PMID- 12850697 TI - Sporadic acute hepatitis E of a 47-year-old man whose pet cat was positive for antibody to hepatitis E virus. AB - We encountered a patient with sporadic acute hepatitis E who had not traveled to areas endemic for hepatitis E virus (HEV) infection and may have been infected in Japan. The patient was a 47-year-old male who had no history of blood transfusion or contact with travelers to hepatitis E-endemic regions or unspecified individuals. The disease presented with general malaise, fever, and brown urine as chief complaints in April 2002. Various hepatitis virus markers were negative, but IgM class antibodies to hepatitis E virus (anti-HEV) and HEV RNA were positive, and the patient was diagnosed with acute hepatitis E. The entire nucleotide sequence (7240 bases) of HEV (HE-JK4) isolated from this patient was determined and compared with known HEV strains. HE-JK4 belonged to genotype IV and exhibited higher similarities to genotype IV HEV strains previously isolated in Japan than to those isolated in China, Taiwan, and Vietnam. The patient's family members living with him were negative for anti-HEV IgG and IgM, but their pet cat was anti-HEV IgG-positive. This finding suggests a potential route of infection of sporadic cases of hepatitis E in Japan. Since the presence of HEV indigenous to Japan is predicted, HEV infection should be considered in the diagnosis of acute hepatitis of unknown cause, even for patients who have not traveled abroad. PMID- 12850698 TI - Ciliated hepatic foregut cyst. A report of one case and a review of the literature. AB - We encountered a patient with a ciliated hepatic foregut cyst (CHFC), which is an uncommon cystic lesion of the liver and hard to distinguish from malignant tumor in imaging features. Cases of CHFC are very rare, five cases were reported in the 19th century and 53 cases in the 20th century. The histogenesis of CHFC is still unclear, but most authors consider that it could arise from the embryonic foregut. A few cases of CHFC mimicking neoplasm were reported. When the diagnosis of CHFC was obtained by fine needle aspiration, close follow-up is necessary in order to find early malignant change. PMID- 12850699 TI - A case of hepatic inflammatory pseudotumor with primary biliary cirrhosis. AB - Inflammatory pseudotumor (IPT) of the liver is an unusual non-neoplastic benign lesion. A 75-year-old man was hospitalized for esophageal varices and gastric cancer. Three years before admission, he had been diagnosed as having primary biliary cirrhosis (PBC) without Sjogren's syndrome. Computed tomography (CT) and magnetic resonance imaging (MRI) scans showed multiple masses (S3, S5, S6) less than 2 cm in diameter in the liver. Since these masses were difficult to distinguish from hepatocellular carcinoma, or metastatic liver carcinoma, one of the masses (S5) was removed during an operation for gastric cancer. Histological examination demonstrated marked infiltration of plasma cells and some histiocytes, findings consistent with the histological features of IPT. The coexistence of hepatic IPT and PBC in this case may have been an accidental event. However, the immunological and environmental factors associated with PBC are thought to be involved in the development of IPT; in addition, cholangitis associated with PBC could have contributed to the development of IPT. PMID- 12850700 TI - Hepatocellular carcinoma associated with secondary haemochromatosis in non cirrhotic liver: a case report. AB - We describe one case of hepatocellular carcinoma (HCC) arising in secondary haemochromatosis in a non-cirrhotic liver. The patient was a 40-year-old male. He had severe pancytopenia due to myelodysplastic syndrome (MDS) and developed secondary haemochromatosis as a result of a large amount of erythrocyte transfusion. Multiple nodules of the liver appeared about 6 years after the diagnosis of MDS. Needle biopsy of the nodules histologically confirmed them to be moderately differentiated HCCs. The liver parenchyma was shown to be non cirrhotic and a deposit of hemosiderin was also identified, consistent with a finding of haemochromatosis of the liver. Transarterial chemoembolization was performed to treat multiple HCCs. There are a number of reports describing HCC occurrence in non-cirrhotic patients with hereditary haemochromatosis. However, HCC in secondary haemochromatosis without cirrhosis is extremely rare. PMID- 12850701 TI - The 6th International Meeting on MR Applications to Porous Media. Ulm, Germany, 8 12 September 2002. Proceedings and abstracts. PMID- 12850702 TI - Giulio Cesare Borgia: founder, promoter, and nurturer of the MRPM Conferences. PMID- 12850703 TI - Magnetic resonance in porous media: forty years on. AB - A personal view of the field of magnetic resonance in porous media is presented in which an attempt is made to survey the current status and achievements, to highlight some of the contributions made by my group over the years and, at the end, to try and identify where further effort and growth points may be perceived. All this is done with the knowledge that the first and last sections are certain to be partial, incomplete and wrong, at least in part, and that the middle section describes work carried out by some of the many excellent students, post doctoral researchers and other colleagues with whom it has been a pleasure to collaborate over a forty year research career. PMID- 12850704 TI - Low-frequency proton NMR relaxometry of a polymer dispersed liquid crystal above TNI. AB - Using proton NMR relaxometry in the kilohertz frequency range, we study dynamics of 5CB liquid crystal molecules dispersed in the form of spherical microdroplets in a PDLC material. The focus of the study is the spin-lattice relaxation in the rotating frame, T1rho(-1), measured above the nematic-isotropic transition TNI. We show that the relaxation rate T1rho(-1)--when induced by uniform molecular translational diffusion in a spherical cavity--depends on the strength of the rotating magnetic field as T1rho(-1) proportional to omega1(-alpha) where alpha varies between 0.7 and 1, depending on the thickness of the ordered surface layer. This relaxation mechanism governs mainly the transverse spin relaxation, whereas the measurements of the frequency and temperature dependence of T1rho(-1) indicate a strong effect of slowing-down of molecular translational diffusion in contact with the polymer surface and yield the average dwell-time of molecules at the surface of the order 10(-5) s. PMID- 12850705 TI - Slow dynamics in colloidal glasses and porous media as probed by NMR relaxometry: assessment of solvent levy statistics in the strong adsorption regime. AB - Mesoscopic media such as porous materials or colloidal pastes develop large specific surface area which strongly influence the dynamics of the embedded fluid. This fluid confinement can be used either to probe the interfacial geometry (frozen porous media) or the particle dynamics (paste and colloidal glass). In the strong adsorption regime, it was recently proposed that the effective surface diffusion on flat surface is anomalous and exhibits long time pathology (Levy walks). This phenomena is directly related to the time and space properties of loop trajectories appearing in the bulk between a desorption and a readsorption step. The Levy statistics extends the time domain of the embedded fluid dynamics toward the low frequency regime. An interesting way to probe such a slow interfacial process is to use field cycling NMR relaxometry. In the first part of this paper, we propose a simple theoretical model of NMR dispersion which only involves elementary time steps of the solvent dynamics near an interface (loops, trains, tails in relation with the confining geometry). In the second part, field cycling NMR relaxometry is used to probe the slow solvent dynamics in two type of interfacial systems: (i) a colloidal glass made of thin and flat particles (ii) two fully saturated porous media, the Vycor glass and MCM48 respectively. Experimental results are critically compared to closed-form analytical expressions and numerical simulations. PMID- 12850706 TI - Structure-mobility relations of molecular diffusion in nanoporous materials. AB - Depending on the measuring conditions, pulsed field gradient (PFG) NMR measurements of molecular diffusion in beds of nanoporous particles may provide information about the propagation rate of guest molecules in both the intra- and interparticle spaces, as well as through the interface between them. Recent progress in both PFG NMR instrumentation and computational techniques have initiated studies of novel aspects in each of these areas, which are reviewed in this communication. They concern the possibility of multicomponent diffusion measurements with ultra-high pulsed field gradients, the peculiarities of molecular diffusion in channel networks, the determination of the surface-to volume ratio of nanoporous particles and the dependence of the tortuosity factor of long-range diffusion on the diffusion mode in the intercrystalline space. PMID- 12850707 TI - Surface nuclear magnetic relaxation and dynamics of water and oil in granular packings and rocks. AB - Low field proton nuclear spin-relaxation at variable magnetic field strength and temperature provides surface dynamical parameters such as surface diffusion coefficients, activation energies, time of residence and coefficient of surface affinity. These parameters were extracted from measurements on grain packs and natural oil-bearing rocks. On grain packs, we show first that changing the amount of surface paramagnetic impurities leads to striking different relationships between the pore-size and the relaxation times T1 and T2. These relationships are well supported by fast-diffusion (surface-limited) or slow-diffusion relaxation models. Surface relaxivity parameters rho1 and rho2 are deduced from the pore size dependence in the fast-diffusion regime. Then, we evidence the frequency and temperature dependence of the surface relaxivity rho1 by field cycling NMR relaxation and relevant theoretical models. The typical frequency dependence found allows an experimental separation of the surface and bulk microdynamics in granular packings and petroleum rocks and the determination of the above mentioned surface dynamical parameters. Finally, we present the first field cycling nuclear spin relaxation experiments performed in water/oil saturated petroleum rocks. We believe that these experiments give new information about the surface localization of these two saturating liquids in pores. PMID- 12850708 TI - Studies of porous media by thermally polarized gas NMR: current status. AB - Three examples of thermally polarized gas NMR performed at New Mexico Resonance are presented to demonstrate its unique advantages in porous media studies. 1) In vivo animal lung imaging by Kuethe et al., in which useful quality 3D images of rat lungs were obtained in 30 min. It is conjectured that comparable human lung images would take much less time to make, possibly by the ratio of body weights, a factor of several hundred. 2) The success of the lung imaging suggested other porous media as candidates for thermally polarized gas NMR. Caprihan and coworkers obtained excellent images from partially sintered ceramics and Vycor glass. Since then, Beyea has developed the technique of spatially resolved BET curves for ceramics and other nanoporous solids. In this way, surface area, pore size, and porosity, averaged over an image voxel, can be spatially resolved. This greatly aids in the characterization of such materials, especially with regards to spatial heterogeneities. 3) Finally, we describe Codd's propagator experiments on propane gas flowing through a packed bed of 300 microm beads. In order to increase signal-to-noise ratio, the flowing gas was pressurized to 170 kPa. Excellent quality propagators showing the discrete nature of the bead pack were obtained. This type of information is not available in comparable liquid studies because most spins will not diffuse far enough to sample the walls in the time available. PMID- 12850709 TI - Novel NMR techniques for porous media research. AB - Diffusion in porous media has been used as a probe of pore geometry in various NMR techniques. We will examine the effect of time-dependent diffusion in CPMG by showing that the diffusion time in CPMG is approximately the echo time, even in grossly inhomogeneous magnetic fields. Extension of the diffusion time in modified CPMG sequences is discussed. Diffusion in the susceptibility-contrast induced internal field is discussed as a means to probe pore size and pore shape. Finally, we present the general concept of two-dimensional relaxation-type experiments for study of molecules, fluids, materials and their dynamics that are characterized by spin relaxation and diffusion. PMID- 12850710 TI - In situ magnetic resonance measurement of conversion, hydrodynamics and mass transfer during single- and two-phase flow in fixed-bed reactors. AB - In recent years there has been increasing interest in applying magnetic resonance (MR) techniques in areas of engineering and chemical technology. The science that underpins many of these applications is the physics and chemistry of transport and reaction processes in porous materials. Key to the exploitation of MR methods will be our ability to demonstrate that MR yields information that cannot be obtained using conventional measurement techniques in engineering research. This article describes two case studies that highlight the power of MR to give new insights to chemical engineers. First, we demonstrate the application of MR techniques to explore both mass transfer and chemical conversion in situ within a fixed bed of catalyst, and we then use these data to identify the rate controlling step of the chemical conversion. Second, we implement a rapid imaging technique to study the stability of the gas-liquid distribution in the low- and high-interaction two-phase flow regimes in a trickle-bed reactor. PMID- 12850711 TI - Pore surface exploration by NMR. AB - A carefully chosen set of experimental techniques applied to porous media characterization provides results that can be much greater than the sum of the individual parts. The inter-relation and complementarity of a number of techniques will be considered. NMR cryoporometry provides a valuable method of pore size measurement. An NMR method that is more widely used to assess pore dimensions relies on relaxation time analysis of a liquid that fills the pores and the enhanced relaxation that occurs in a liquid at the solid/liquid interface. Thermoporometry, a method based on the application of Differential Scanning Calorimetry (DSC), is closely related to cryoporometry, but employs a different set of assumptions to evaluate pore size distributions. Comparison of the results obtained on the same samples using all these methods together with gas adsorption serves to validate the methods and provide significantly more information about surface-fluid interaction and the behavior of nano-scale material within pores than each method employed in isolation. Technique developments will be discussed and applications of these methods to ideal silicas will be used to illustrate their power, particularly in combination. PMID- 12850712 TI - Bone tissue and porous media: common features and differences studied by NMR relaxation. AB - Despite significant differences between bone tissues and other porous media such as oilfield rocks, there are common features as well as differences in the response of NMR relaxation measurements to the internal structures of the materials. Internal surfaces contribute to both transverse (T2) and longitudinal (T1) relaxation of pore fluids, and in both cases the effects depend on, among other things, local surface-to-volume ratio (S/V). In both cases variations in local S/V can lead to distributions of relaxation times, sometimes over decades. As in rocks, it is useful to take bone data under different conditions of cleaning, saturation, and desaturation. T1 and T2 distributions are computed using UPEN. In trabecular bone it is easy to see differences in dimensions of intertrabecular spaces in samples that have been de-fatted and saturated with water, with longer T1 and T2 for larger pores. Both T1 and T2 distributions for these water-saturated samples are bimodal, separating or partly separating inter- and intratrabecular water. The T1 peak times have a ratio of from 10 to 30, depending on pore size, but for the smaller separations the distributions may not have deep minima. The T2 peak times have ratios of over 1000, with intratrabecular water represented by large peaks at a fraction of a ms, which we can observe only by single spin echoes. CPMG data show peaks at about a second, tapering down to small amplitudes by a ms. In all samples the free induction decay (FID) from an inversion-recovery (IR) T1 measurement shows an approximately Gaussian (solid-like) component, exp[-1/2 (T/TGC), with TGC approximately 11.7+/ 0.7 micros (GC for "Gaussian Component"), and a liquid-like component (LLC) with initially simple-exponential decay at the rate-average time T(2-FID) for the first 100 micros. Averaging and smoothing procedures are adopted to derive T(2 FID) as a function of IR time and to get T1 distributions for both the GC and the LLC. It appears that contact with the GC, which is presumed to be 1H on collagen, leads to the T2 reduction of at least part of the LLC, which is presumed to be water. Progressive drying of the cleaned and water-saturated samples confirms that the long T1 and T2 components were in the large intertrabecular spaces, since the corresponding peaks are lost. Further drying leads to further shortening of T2 for the remaining water but eventually leads to lengthening of T1 for both the collagen and the water. After the intertrabecular water is lost by drying, T1 is the same for GC and LLC. T(2-FID) is found to be roughly 320/alpha micros, where alpha is the ratio of the extrapolated GC to LLC, appearing to indicate a time tau of about 320 micros for 1H transverse magnetization in GC to exchange with that of LLC. This holds for all samples and under all conditions investigated. The role of the collagen in relaxation is confirmed by treatment to remove the mineral component, observing that the GC remains and has the same TGC and has the same effect on the relaxation times of the associated water. Measurements on cortical bone show the same collagen related effects but do not have the long T1 and T2 components. PMID- 12850713 TI - Magnetic resonance profiling studies of the drying of film-forming aqueous dispersions and glue layers. AB - We report magnetic resonance profiling experiments to monitor (i) the drying of alkyd emulsion layers, (ii) the cure of wood glue layers and (iii) water transport through glue lines. The alkyd drying is a two stage process. We report new results which support previous evidence that the alkyd drops do not coalesce until the water fraction is below circa 0.02. The profiles recorded from glue layers suggest that MR is a sensitive probe of the curing process and barrier properties of the glue. The measurements were made using GARField (stray field, STRAFI like) magnetic resonance profiling and an improved GARField magnet design characterized by two values of the gradient-to-field strength ratio at two locations offering the same field strength is also reported. PMID- 12850714 TI - Use of the second dimension in PGSE NMR studies of porous media. AB - 2-dimensional methods based on PGSE NMR may be used to correlate or separate molecular dynamical properties, or to elucidate fluctuations. These may utilize either the gradient (q-vector) domain, in which molecular displacements are measured, or the time domain, in which relaxation is measured, and may be analyzed by combinations of inverse Fourier or Laplace transforms. Existing methodologies are reviewed and new experiments proposed. In particular the use of diffusion-diffusion exchange and correlation analysis is demonstrated using the case of water diffusion in a lamellar phase liquid crystal. PMID- 12850715 TI - Analysis of historical porous building materials by the NMR-MOUSE. AB - Samples of sandstone with and without deposits of silicon oxide stone strengthener as well as samples of historical brick material were analyzed by transverse NMR relaxation and mercury intrusion porosimetry. Relaxation times and relaxation time distributions of the protons from the water saturated samples were measured by low-field NMR using homogeneous and inhomogeneous fields. The measurements in inhomogeneous fields were performed with two different NMR-MOUSE sensors, one with a field gradient of 2 T/m and the other with an average field gradient of about 20 T/m. In the sandstone samples the application of stone strengtheners was shown to result in a confinement of the large pores within the outer layer of a few millimeters depth. Depending on the ferromagnetic contamination of the brick samples, the relaxation time distributions can be affected. The agreement of T2 relaxation time distributions and pore size distributions from mercury intrusion porosimetry was found to be better for the NMR-MOUSE sensors than for the homogeneous field measurements. This is true even for different brick samples, unless the content in ferromagnetic particles is very strong. PMID- 12850716 TI - A new probe of pore geometry: diffusive MASS NMR. AB - NMR methods are widely used to probe the structure and fluid dynamics of porous materials, as they are uniquely suited to these studies since NMR records the correlation of changing local magnetic fields over a time scale of ns to seconds. The local magnetic fields are established by local variations in the bulk magnetic susceptibility of the sample (and so are directly tied to the sample's local structure). The fluctuation in field that a spin sees is due to molecular transport (including molecular diffusion) through these local fields, and so reports on the length scales of structures and impediments to transport. We have developed a new set of methods DIFFUSIVE-MASS to provide a means of systematically varying the effective time scale of the measurement and thus the effective length scale. This new handle permits a detailed, microscopic picture of the structure and dynamics. Diffusive MASS NMR methods will permit a systematic set of methods and analysis for characterizing the chemistry, structure and fluid dynamics of the mobile phase in porous materials. The approach will be applicable to any diamagnetic material. In particular, the industry of oil discovery depends on understanding heterogeneous porous media. PMID- 12850717 TI - Molecular mobility in fixed-bed reactors investigated by multiscale NMR techniques. AB - The complex problem of a fixed-bed reactor consisting of catalytically active particles provides an exceptional opportunity of combining a wide range of NMR methods which have become available over time as tools to probe porous media. This work demonstrates the feasibility of different NMR techniques for the investigation of the intra- and interparticle pore space over length scales from nanometers up to centimeters. Many industrially relevant cracking reactions leave a coke residue on the inner surface of the porous catalyst particles so that the active sites become inaccessible to the reactants. Moreover, the pore space shrinks due to the formation of coke, thereby hindering molecular transport. The presence of the coke residue and its influence on the mobility of adsorbed fluid molecules are probed by 129Xe spectroscopy, NMR cryoporometry, relaxation dispersion measurements, and investigations of the reduced diffusivity in the intraporous space. The voids surrounding the random arrangement of catalyst pellets represent another pore space of much larger dimensions, the properties of which can be more directly investigated by mapping the fluid density and the velocity distribution from velocity-encoded imaging. Propagator representations averaged over large sample volumes are discussed and compared to velocity images obtained in selected axial slices of the reactor. PMID- 12850718 TI - NMR properties of petroleum reservoir fluids. AB - NMR well logging of petroleum reservoir require the measurement of the NMR response of water, oil, and gas in the pore space of rocks at elevated temperatures and pressures. The viscosity of the oil may range from less than 1 cp to greater than 10,000 cp. Also, the oil and gas are not a single component but rather a broad distribution of components. The log mean T1 and T2 relaxation time of dead (gas free) crude oils are correlated with viscosity/temperature and Larmor frequency. The relaxation time of live oils deviate from the correlation for dead crude oils. This deviation can be correlated with the methane content of the oil. Natural gas in the reservoir has components other than methane. Mixing rules are developed to accommodate components such as ethane, propane, carbon dioxide, and nitrogen. Interpretation of NMR logs uses both relaxation and diffusion to distinguish the different fluids present in the formation. Crude oils have a broad spectrum of components but the relaxation time distribution and diffusion coefficient distribution are correlated. This correlation is used to distinguish crude oil from the response of water in the pores of the rock. This correlation can also be used to estimate viscosity of the crude oil. PMID- 12850719 TI - Modified oscillating gradient pulses for direct sampling of the diffusion spectrum suitable for imaging sequences. AB - A variation of the oscillating gradient spin echo method had been developed, which isolates temporal frequencies of the dephasing spectrum. This allows sampling of the diffusion spectrum, the Fourier transform of the velocity correlation function (VCF). It has been shown that restriction and flow alter this function in ways that can be mathematically characterized, yielding quantitative information on restriction geometry and flow parameters. It is demonstrated that in many systems of interest, dispersion of velocity will produce a peak in the VCF spectrum near omega=0, while restricted diffusion will manifest itself in the spectrum at higher frequencies. The method, therefore, may be useful for decoupling their effects on the apparent diffusion coefficient (ADC), as well as in revealing the physics of both phenomena. This method has been implemented in model systems of packed beads, yielding data consistent with theoretical models of restricted diffusion spectra and data from one previous study. The method may have significant application to biology and medicine, as well as the study of transport phenomena in porous media and complex flow. PMID- 12850720 TI - Applications of controlled-flow laser-polarized xenon gas to porous and granular media study. AB - We report initial NMR studies of continuous flow laser-polarized xenon gas, both in unrestricted tubing, and in a model porous media. The study uses Pulsed Gradient Spin Echo-based techniques in the gas-phase, with the aim of obtaining more sophisticated information than just translational self-diffusion coefficients. Pulsed Gradient Echo studies of continuous flow laser-polarized xenon gas in unrestricted tubing indicate clear diffraction minima resulting from a wide distribution of velocities in the flow field. The maximum velocity experienced in the flow can be calculated from this minimum, and is seen to agree with the information from the complete velocity spectrum, or motion propagator, as well as previously published images. The susceptibility of gas flows to parameters such as gas mixture content, and hence viscosity, are observed in experiments aimed at identifying clear structural features from echo attenuation plots of gas flow in porous media. Gas-phase NMR scattering, or position correlation flow-diffraction, previously clearly seen in the echo attenuation data from laser-polarized xenon flowing through a 2 mm glass bead pack is not so clear in experiments using a different gas mixture. A propagator analysis shows most gas in the sample remains close to static, while a small portion moves through a presumably near-unimpeded path at high velocities. PMID- 12850721 TI - MRI studies of the evaporation of a single liquid droplet from porous surfaces. AB - A combination of in situ one-dimensional profiling and two-dimensional 1H magnetic resonance imaging has been applied to study the shape and subsequent dynamic evaporation behavior of a single liquid droplet after impact onto a porous surface in a carefully controlled atmosphere. Aspects of the porous media permeability and particle shape are shown to affect considerably the observed drying regime. Variation in the shape, spread ratio, and penetration depth of the liquid droplet following impact are attributed to the different types of porous media used. A comparison of numerical simulations with the MRI data for the droplet shape, spread factor, and penetration depth show good agreement though larger deviations between simulated and experimental results occur in certain cases where the surface of the porous medium is disrupted by the impact of the liquid droplet. Initial results examining the effect of a reduced liquid surface tension show a significantly increased evaporation rate. PMID- 12850722 TI - NMR magnetization transfer as a tool for characterization of nanoporous materials. AB - The application of nuclear magnetic resonance magnetization transfer experiments to probe the surface-to-volume ratio and pore morphology of porous materials with characteristic pore sizes of 1-100 nm is described. The method is based on the phenomenon of incomplete freezing of liquids in small pores where a few monolayers adjacent to the pore walls remain liquid. Sufficient difference between the transverse relaxation times in the solid frozen core and liquid surface layer allows the initial preparation and subsequent re-equilibration of a solid-liquid magnetization grating. The method is demonstrated using model nanoporous materials with known characteristics. The ensuing problems of the mechanism of the magnetization transfer through the interface and within the frozen core are discussed and elucidated by pulsed-field-gradient NMR experiments. PMID- 12850723 TI - Diffusion-relaxation distribution functions of sedimentary rocks in different saturation states. AB - We present diffusion-relaxation distribution functions measured on four rock cores that were prepared in a succession of different saturation states of brine and crude oil. The measurements were performed in a static gradient field at a Larmor frequency of 1.76 MHz. The diffusion-relaxation distribution functions clearly separate the contributions from the two fluid phases. The results can be used to identify the wetting and non-wetting phase, to infer fluid properties of the phases, and to obtain additional information on the geometrical arrangement of the phases. We also observe effects due to restricted diffusion and susceptibility induced internal gradients. PMID- 12850724 TI - Internal magnetic gradient fields in glass bead packs from numerical simulations and constant time diffusion spin echo measurements. AB - Internal magnetic field gradients in water saturated glass bead packs were studied by numerical simulations and a constant time spin echo (CTSE) experiment. The CTSE is comprised of two spin echo refocusing periods where each of the two evolution periods, tau1 and tau2, is varied so that the total evolution, 2(tau1 + tau2), is held constant. The experiment is similar to that introduced by Norwood and Quilter and allows the effects of dephasing due to diffusion in a magnetic field gradient to be separated from other relaxation mechanisms. In our experiments, the magnetic susceptibility difference between the pore fluid and glass beads creates the internal field gradient. CTSE measurements were performed at 7 T (300 MHz 1H) for water saturated in 50 microm diameter glass bead pack. We find that the internal gradients in the center of the pore bodies, where free diffusion applies, is in the range of 10 to 100 G/cm. This fluid volume accounts for < or =10% of the total pore volume. From direct numerical simulations of the internal magnetic field based on a first principles calculation, we find that the major fraction, >90%, of the pore volume has internal gradients of order 500 to 5,000 G/cm. Signals from water in these large gradients is not observed in our CTSE measurements. PMID- 12850725 TI - Salt transport and crystallization in porous building materials. AB - Salt weathering is a major cause of deterioration of porous building materials. To obtain information about the mechanisms underlying these damage processes we have studied the moisture and ion transport. We measured the time evolution of NaCl saturated samples of fired-clay brick during one-sided drying using Nuclear Magnetic Resonance. The moisture content and amount of dissolved Na ions could be measured quantitatively as a function of position. The NaCl concentration profiles obtained from these data reflect the competition between advection to the surface and redistribution by diffusion. By representing the measured moisture and NaCl profiles in an efflorescence pathway diagram (EPD) also the crystallization can be taken into account. PMID- 12850726 TI - Electric field driven flow in natural porous media. AB - Electric fields were applied to fluid-saturated packed sand beds (0.23+/-0.03 mm average pore diameter), and the effects on the mobility of the water molecules were monitored using stimulated echo (STE) and pulsed field gradient (PFG) experiments. The mean flow velocity, averaged over the entire sample, is expected to vanish in closed systems, but the PFG and time dependent signal decay was enhanced beyond the effects of thermal diffusion, due to velocity dispersion. The internal flow generated by the electric field was shown to be fully time reversible upon inverting the electric field polarity (for total flow times of up to 0.4s), a strong indication that the NMR detected displacements were mainly due to electro-osmotic flow (EOF). However, a comparison of the velocity dispersion for different electrolyte concentrations showed that the measured effect scaled with the applied power VI (V = voltage, I = electric current), rather than with the voltage alone, contrary to the prediction of the basic model for EOF in a single capillary channel. PMID- 12850727 TI - Time-resolved observation of the decomposition process of N,N,N Trimethylanilinium cations on zeolite H-Y by in situ stopped-flow 13C MAS NMR spectroscopy. AB - N,N,N-Trimethylanilinium cations were synthesized on acidic zeolite H-Y by direct reaction of N,N-dimethylaniline and methanol. Time-resolved observation of the detailed decomposition process of N,N,N-trimethylanilinium cations was achieved by in situ 13C stopped-flow (SF) MAS NMR spectroscopy at reaction temperatures of 498 K to 573 K. PMID- 12850728 TI - NMR study on the early stages of hydration of a porous carbonate stone. AB - Nuclear Magnetic Resonance was employed to obtain information on the pore filling during the absorption process. A porous carbonate stone, largely employed for buildings and mainly outdoor decorations was studied during water absorption by capillary rise, and filled pores radii were evaluated by comparison between experimental and theoretical parametric magnetization decay curves. Non mono exponential T2 allowed spin populations to be split among the associated different relaxation times. PMID- 12850729 TI - The NMR microimaging studies of the interplay of mass transport and chemical reaction in porous media. AB - PFG NMR is employed to perform a comparative study of the filtration of water and propane through model porous media. It is shown that the dispersion coefficients for water are dominated by the holdup effects even in a bed of nonporous glass beads. It is demonstrated that correlation experiments such as VEXSY are applicable to gas flow despite the large diffusivity values of gases. The PFG NMR technique is applied to study the gravity driven flow of liquid-containing fine solid particles through a porous bed. The NMR imaging technique is employed to visualize the propagation of autocatalytic waves for the Belousov-Zhabotinsky reaction which is carried out in a model porous medium. It is demonstrated that the wave propagation velocity decreases as the wave crosses the boundary between the bulk liquid and the flooded bead pack. The images detected during the catalytic hydrogenation of alpha-methylstyrene on a single catalyst pellet at elevated temperatures have revealed that the reaction and the accompanying phase transition alter the distribution of the liquid phase within the pellet. PMID- 12850730 TI - MRI observation of heavy metal transport in aquifer matrices down to sub-mg quantities. AB - In this contribution, we report results from MRI studies of the propagation of heavy metal ions through columns packed with sandy aquifer materials. Both sorption of low concentrations of heavy metal ions from water flowing through the column and the formation of sorption traces produced from initially localized higher concentrations of heavy metal ions, and the remobilisation of adsorbed ions with time and changing hydrochemical conditions were studied. Multislice spin-echo and FLASH techniques were used for the imaging experiments. Advantages and problems of the different imaging protocols for the study of aquifer materials are discussed. PMID- 12850731 TI - Three-dimensional snow images by MR microscopy. AB - MR microscopy technique was introduced to visualize and quantify the three dimensional structure of snowpack. Since the NMR signal from the ice was week, we looked at the air space instead filling with dodecane or aniline doped with iron acetylacetonate. Four types of snow were tested: ice spheres, large rounded poly crystals, small rounded mono-crystals and depth hoar crystals. A specific specimen-cooling system was developed to keep the temperature below 0 degrees C. In the experiments 0.5 to 2 h were necessary to accumulate the signals enough to obtain a 3D micro-image; the image matrix 128(3), voxel size (200 microm)3 or 256(3) (120 microm)3. Comparison with the 2D data using the conventional section plane method was also carried out and MR microscopy is proved to be a very useful method to visualize the microstructure of snowpack. PMID- 12850733 TI - Quantitative 'real-time' imaging of multi-phase flow in ceramic monoliths. AB - An extension of the RARE technique has been developed which acquires multiple images from a single radio-frequency excitation. This pulse sequence has been used to image, in real-time, gas flow through stagnant liquid within parallel channel ceramic monoliths. From these images, gas-phase volume fractions, and distributions of gas bubble length and velocity as a function of gas flow rate (50-300 cm3 min(-1)) and channel size (300 and 400 channels per square inch, cpsi) are obtained directly. Increasing the gas flow rate increased the number of large bubbles and the average bubble velocity. A bimodal distribution in the bubble velocities was observed for flow within the larger channel size (300 cpsi) in contrast to a broad unimodal distribution characterizing two-phase flow within the smaller channel size (400 cpsi). PMID- 12850732 TI - What is the surface specific area of porous cement-based material? A nuclear magnetic relaxation dispersion approach. AB - We propose a new NMR method to measure and follow the evolution of the surface specific area, Sp, of a porous cement-based material. This method, that does not require any preliminary drying process, uses nuclear magnetic relaxation dispersion (NMRD), the measurement of spin-lattice relaxation rate as a function of magnetic field strength or nuclear Larmor frequency. The method is applied for three different mortars samples prepared by mixing cement, sand, silica fume, water and superplasticizer with a water to cement ratio w/c=0.25, 0.38 and 0.65, respectively. The evolution of Sp grows linearly with the degree of advancement of chemical reactions measured by thermal heating and we evidence two relaxation processes independent of the w/c ratio. PMID- 12850734 TI - Diffusion studies in porous media with the "inside-out" technique. AB - Diffusion measurements of water in a glass beads porous media were carried out by the "One-Sided Access" sensor. An approach to determine the long time mean square displacement of hydrogen molecules by measuring the ratio of stimulated echo to primary echo in a three pulse sequence in the presence of permanent static field gradient is analysed. The difference between the apparent diffusion coefficient in a pure water sample and in water/glass beads mixtures becomes visible. PMID- 12850735 TI - Magnetic resonance imaging study of complex fluid flow in porous media: flow patterns and quantitative saturation profiling of amphiphilic fracturing fluid displacement in sandstone cores. AB - Magnetic resonance imaging is used to follow the removal process of a visco elastic surfactant (VES) fracturing fluid in Bentheimer sandstone cores at typical reservoir temperatures (T=333 K). Two displacing fluids were investigated, a Gadolinium doped water phase (1M NaCl solution), and a Gadolinium doped hydrocarbon phase (Mineral Spirits). In addition to flow characteristics obtained by conventional core-flooding, i.e., the macroscopically averaged volumetric flow rates and differential pressures, we have also measured the saturation profiles and characteristic displacement patterns during all stages of the removal process. To acquire these data we have used quantitative one dimensional chemically specific profiling along with fast two-dimensional imaging experiments while flooding Bentheimer sandstone cores in situ in the spectrometer. Our results show that both displacement processes (complex fluid displaced by water or hydrocarbon phase) are dominated by the large viscosity contrasts present. However, distinct differences were found between the displacement characteristics of water and hydrocarbon, which confirmed the sensitivity of the complex fracturing fluid to the displacing fluid. PMID- 12850736 TI - Micropore size analysis by NMR in hydrated cement. AB - The understanding of the microstructure of cement remains incomplete. Especially, the progressive setting of the material is still unclear. Micropore size distribution (microstructure) has been investigated by both standard proton nuclear magnetic relaxation (1H-NMR) and field-cycling relaxation in C3S hydrated paste. The non-exponential decay was interpreted as a distribution of discrete relaxation rates. The attribution of T1 is supported by both a spectral and a dispersion curve analyses. These experiments allow us to follow the structuration of the material during setting. PMID- 12850737 TI - Spatial dependence of dispersion. AB - The use of Pulsed Gradient Spin Echo (PGSE) NMR to investigate flow in porous media is well established. Using two pairs of position-encoding pulses in the PGSE experiment gives the possibility of examining velocity fluctuations, by comparing displacements, during the two encoding intervals. This method may be used to measure the asymptotic dispersion coefficient as well as the Velocity Auto-Correlation Function (VACF) in porous media flow. Some examples of two dimensional maps of density and velocity distributions are accompanied by the first attempt to perform localized measurements of flow dispersion in porous media. PMID- 12850738 TI - Flow of newtonian/non-newtonian fluids in a bundle of tubes and in a packing of beads by MRI. PMID- 12850739 TI - 2D relaxation/diffusion correlations in porous media. AB - 2D correlations between NMR relaxation and/or diffusion have been used to investigate water and oil dynamics in food and micro-emulsion systems. In the case of Mozzarella and Gouda cheese samples, a significant change in D/T2 correlation is appearing with cheese aging. In the case of a water/toluene micro emulsion, some evidence for coalescence effects is suggested by D/D exchange spectra. PMID- 12850740 TI - Quantitative evaluation of porous media wettability using NMR relaxometry. AB - We propose a new method to determine wettability indices from NMR relaxometry. The new method uses the sensitivity of low field NMR relaxometry to the fluid distribution in oil-water saturated porous media. The model is based on the existence of a surface relaxivity for both oil and water, allowing the determination of the amount of surface wetted either by oil or by water. The proposed NMR wettability index requires the measurement of relaxation time distribution at four different saturation states. At the irreducible water saturation, we determine the dominant relaxation time of oil in the presence of a small amount of water, and at the oil residual saturation, we determine the dominant relaxation time of water in the presence of a small amount of oil. At 100% water and 100% oil saturation, we determine the surface relaxivity ratio. The interaction of oil with the surface is also evidenced by the comparison of the spin-lattice (T1) and spin-locking (T1rho) relaxation times. The new NMR index agrees with standard wettability measurements based on drainage-imbibition capillary pressure curves (USBM test) in the range [-0.3-1]. PMID- 12850741 TI - Investigation of two phase flow and phase trapping by secondary imbibition within Fontainebleau sandstone. AB - Pulsed magnetic field gradient stimulated echo NMR is used to investigate the simultaneous flow of two phases (an aqueous phase and an hydrocarbon phase) within a strongly water-wet sample of Fontainebleau sandstone. The Fontainebleau sandstone is prepared in increasing steady-state water saturations by a secondary imbibition process. The increase in the water saturation causes an increasing fraction of the oil phase (non-wetting phase) to become trapped within the sample. The stimulated echo dependence on the gradient pulse area, q, is used to derive the displacement probability, PX, for a fixed observation time. These displacement probabilities clearly show the progressive trapping of the hydrocarbon phase with increasing steady-state water saturations. Quantitative measurements of the fraction of the oil phase trapped were made from the echo attenuation function Edelta(q), both as a function of water saturation and observation time. PMID- 12850742 TI - Improved pore space structure characterization by fusion of relaxation tomography maps. AB - Quantitative Relaxation Tomography in porous media furnishes maps of internal sections where each pixel represents T1 or T2 of water 1H in the corresponding voxel, so that quantitative information on the pore space structure can be obtained. The porosity can be determined at different length scales by correcting pixel by pixel the signal intensity for T2 decay. Moreover, on the basis of the distribution of T1, the microporosity fraction can be computed, as well as several voxel-average porosities. Since T1 and T2 encode different pieces of information, fusion image techniques can improve the characterization of the pore space, showing simultaneously, on the same image, maps of the two parameters. Examples are given of application to a water-saturated travertine core and to a pig femur. Different kinds of look-up tables were tried by varying two of the three dimensions of the HSV color space in such a way as to optimize both the T1 and T2 contrasts simultaneously. PMID- 12850743 TI - Magnetic resonance studies of dissolving particulate solids. AB - Magnetic resonance methods have been used to elucidate the internal pore structure of particulate solids, in particular detergent tablets. Such information is essential to a comprehensive understanding of the dissolution characteristics of these materials and how this property is related to processing conditions during tablet formation. In particular 3-D images of porosity are produced and 2-D self-diffusion maps are acquired as a function of observation time, which enables pore size to be quantified as a function of position via the extracted surface-to-volume ratio of the pore space. These properties are determined as a function of processing parameters, in particular the compression force used in tablet formation. PMID- 12850744 TI - Single sided imaging sensor. AB - A portable single sided sensor to render depth profiles of MR signal non invasively is presented. The device utilizes a tailored permanent magnet array producing flat sensitive volumes up to 40 mm from the mobile inspection head. Automated slice selection is performed using a set of switched capacitors in the tuning and receiver circuits. A single channel spectrometer drives the multi slice acquisition. The RF coil and magnet array geometries have been tailored to house gradient sets to further extend the image rendering capabilities for in plane spatial resolution (2D and 3D MRI). The probe has been tested acquiring moisture profiles in concrete and porous rocks and measuring diffusion parameters on a phantom sample. PMID- 12850745 TI - The study of Xe adsorption behavior in meso-size pores of carbon black materials using laser-polarized 129Xe NMR spectroscopy. AB - The meso size pores of carbon black materials with Pt critically affect catalysts which play an important role for fuel cells of electric vehicles. Time-consuming BET methods are usually used to measure the physisorption enthalpy which determines the characteristics of catalysts. The laser polarized method enhances 129Xe polarization by 4 orders of magnitude, overcoming a low sensitivity, making this measurement technique faster than conventional experiments. In this paper, we first demonstrate Laser-Polarized 129Xe NMR Spectroscopy for studying carbon black materials with Pt of fuel cells of electric vehicles in order to determine the physisorption enthalpy. At the same time, T1 experiments using Laser Polarized 129Xe will be discussed in order to clarify the surface condition and adsorption behavior. PMID- 12850746 TI - Deformation and failure of rock samples probed by T1 and T2 relaxation. AB - The objective of the study was to pinpoint the effect of stress induced rock matrix alterations on NMR-wireline-log measurements by means of laboratory T1 and T2 relaxation time measurements. The research activities were subdivided into two major parts: NMR relaxation measurements on a brine saturated outcrop sandstone (Red Wildmoor Sandstone) during uniaxial compressional tests and NMR relaxation measurements on artificial sandstone samples prepared with defined crack patterns. T1-measurements performed on Red Wildmoor samples during compaction showed a decrease in the mean relaxation rate 1/exp