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T Beck

Publications and source records attributed to T Beck.

At least 19 recordsLinked to original sources

Development of conventional and real-time NASBA for the detection of Legionella species in respiratory specimens.

Isothermal nucleic acid sequence-based amplification (NASBA) was applied to detect Legionella 16S rRNA. The assay was originally developed as a Legionella pneumophila conventional NASBA assay with electrochemiluminescence (ECL) detection and was subsequently adapted to a L. pneumophila real-time NASBA format and a Legionella spp. real-time NASBA using molecular beacons. L. pneumophila RNA prepared from a plasmid construct was used to assess the analytical sensitivity of the assay. The sensitivity of the NASBA assay was 10 molecules of in vitro wild type L. pneumophila RNA and 0.1-1 colony-forming units (CFU) of L. pneumophila. In spiked respiratory specimens, the sensitivity of the NASBA assays was 1-10000 CFU of L. pneumophila serotype 1 depending on the background. After dilution of the nucleic acid extract prior to amplification, 1-10 CFU of L. pneumophila serotype 1 could be detected with both detection methods. Finally, 27 respiratory specimens, well characterized by culture and PCR, collected during a L. pneumophila outbreak, were tested by conventional and real-time NASBAs. All 11 PCR positive samples were positive by conventional NASBA, 9/11 and 10/11 were positive by L. pneumophila real-time NASBA and Legionella spp. real-time NASBA, respectively.

Bile↗

Non-invasive coronary angiography with 16-slice spiral computed tomography: image quality in patients with high heart rates.

The purpose of this study was to assess segment image quality at high heart rates using 16-slice computed tomography and differential reconstruction for major coronary vessels. According to the following protocol, 16-slice CT coronary angiography in 46 patients with a mean heart rate of 86.3+/-11.8 was reconstructed. At three transverse planes, preview series were obtained and motion artifacts evaluated in 5% increments from 0-95% within the cardiac cycle. Relying on image quality in the previews, reconstructions were performed at three z-positions for each patient. Segment image quality was assessed in terms of artifacts and visibility. The effects of heart rate and trigger delay on image quality were analyzed. Optimal image quality was achieved at 25 to 35% of the cardiac cycle for the left circumflex (CX) and right coronary artery (RCA) or 30 to 40% for the left main (LM) and left anterior descending artery (LAD). Sixteen-slice CT and differential reconstruction produced good image quality with a low percentage of motion-degraded proximal and middle segments (8.8%). Grades were 1.5 for the LM, 1.9 for the LAD, 2.0 for the CX and 2.3 for the RCA. At high heart rates, good image quality of the coronary arteries is achieved by 16-slice CT and a sophisticated reconstruction strategy at peak to late systole.

Algorithms↗

Image quality and diagnostic accuracy of 16-slice multidetector computed tomography for the detection of coronary artery disease in obese patients.

BACKGROUND: Cardiac multislice spiral computed tomography (MSCT) scanners permit visualization of the coronary arteries with an overall good sensitivity (sens) and specificity (spec). However, in obese patients (pts), who are at higher risk to develop coronary artery disease (CAD), image quality of MSCT is supposed to be limited. At present, there are no data whether the accuracy of MSCT depends on the body mass index (BMI). Thus, we compared the catheter-controlled MSCT results from normal weight and obese pts in a cohort of 117 pts with regard to sens, spec, positive predictive value (PPV), negative predictive value (NPV) and image quality. METHODS AND MATERIAL: In all, 21 normal weight pts (group I: BMI<25, 64.6+/-11.1 years, number of risk factors 2.1+/-1.1), 60 pts with mild overweight (group II: BMI 25-30, 64.6+/-8.9 years, number of risk factors 3.4+/-1.0) and 36 obese pts (group III: BMI >30, 63.0+/-8.5 years, number of risk factors 3.4+/-0.9) were examined by MSCT (Sensation 16 Speed 4 D((R)), Siemens, Germany, gantry rotation time 375 ms) and invasive coronary angiography. MSCT results were compared blinded to the results of the coronary angiography with regard to the presence or absence of a significant stenosis (>50%) in a modified AHA 13 segment (sgt) model. Image quality was assessed on a qualitative scale between 1 (very good) and 5 (insufficient image quality) for each sgt. RESULTS: Sens, spec, PPV and NPV were statistically not different in all three groups (I: 0.88/0.97/0.91/0.96, II: 0.83/0.97/0.88/0.95, III: 0.87/0.99/0.96/0.96). 3 pts (group I 1, group II 2) had to be excluded from analysis due to technical problems. Group I had significantly less risk factors (P < 0.001) and image quality was significantly better than in group II and III (P < 0.05). Group II and III did not differ with regard to risk factors or image quality. CONCLUSIONS: Overweight and obesity have an impact on MSCT image quality but did not hamper the diagnostic accuracy. Thus, MSCT is a noninvasive method to detect or rule out CAD also in pts with higher BMI. These retrospective data have to be confirmed in larger prospective trials.

Aged↗

Two quality control exercises involving nucleic acid amplification methods for detection of Mycoplasma pneumoniae and Chlamydophila pneumoniae and carried out 2 years apart (in 2002 and 2004).

The quality performance of laboratories for the detection of Mycoplasma pneumoniae and Chlamydophila pneumoniae by two quality control (QC) exercises with a 2-year interval was investigated. For the 2002 QC exercise, specimens were spiked with M. pneumoniae at concentrations of 5,000, 500, 50, and 0 color-changing units (CCU)/100 microl. The limit of detectability was 50 CCU/100 microl. Therefore, this concentration was omitted from the 2004 panel and was excluded from the analysis. In 2002, 2 out of 12 participants obtained 100% correct results, 2 out of 12 produced false-positive results, and 10 out of 12 had between 0 out of 9 and 8 out of 9 correct positive results. In 2004, correct results were obtained in 15 out of 18 tests, and no false-positive results were reported. In 2002, specimens were spiked with C. pneumoniae at concentrations of 490, 49, 4.9, and 0 inclusion-forming units/100 microl (IFU/100 microl). In the 2004 panel, samples spiked with a lower dilution of 0.49 IFU/100 microl were added to the panel. For the C. pneumoniae QC, correct results were produced in 12 out of 16 and 13 out of 18 tests in 2002 and in 2004, respectively. Both multiplex PCR and nucleic acid sequence-based amplification (NASBA) formats scored a smaller number of samples positive than the monoplex reactions.

Bacteriological Techniques↗

Development of conventional and real-time nucleic acid sequence-based amplification assays for detection of Chlamydophila pneumoniae in respiratory specimens.

Isothermal nucleic acid sequence-based amplification (NASBA) was applied to the detection of Chlamydophila pneumoniae 16S rRNA by using the NucliSens basic kit (bioMérieux, Boxtel, The Netherlands). The assay was originally developed as a conventional NASBA assay with electrochemiluminescence detection and was subsequently adapted to a real-time NASBA format by using a molecular beacon. C. pneumoniae RNA prepared from a plasmid construct was used to assess the analytical sensitivity of the assay. The sensitivity of the NASBA assay was 10 molecules of in vitro wild-type C. pneumoniae RNA and 0.1 inclusion-forming unit (IFU) of C. pneumoniae. In spiked respiratory specimens, the sensitivity of the C. pneumoniae NASBA assay varied between 0.1 and 1 IFU/100 mul sample, depending on the type of specimen. Finally, conventional and real-time NASBA were applied to respiratory specimens previously tested by PCR. A 100% concordance between the test results was obtained.

Chlamydophila pneumoniae↗

Clinical use of multislice spiral computed tomography in 210 highly preselected patients: experience with 4 and 16 slice technology.

OBJECTIVE: To report an initial experience with multislice spiral computed tomography (MSCT) coronary imaging, as well as differences in diagnostic accuracy between 4 slice and 16 slice MSCT technology. METHODS AND RESULTS: 210 patients underwent MSCT coronary angiography (4 slices, n = 120; 16 slices, n = 90; suspicion of coronary artery disease, n = 158; suspicion of restenosis, n = 52). Recommendations for further diagnostic tests were based on the MSCT results. Patients were interviewed by telephone after a mean (SD) of 449 (169) days to evaluate their further clinical course. MSCT detected significant lesions in 90 of 210 (43%) patients and invasive coronary angiography (ICA) was recommended. MSCT excluded significant lesions in 120 of 210 (57%) patients. ICA was actually performed in 44 of 210 (21%) patients (corresponding results, 27 of 44 (61%); false positive, 11 of 44 (25%); false negative, 6 of 44 (14%)). No significant differences were found between 4 and 16 slice imaging. No major cardiac event occurred during follow up. CONCLUSIONS: MSCT was found to be useful to evaluate the need for invasive diagnostic procedures. However, the false negative results underline that further improvements of image quality are required before MSCT can replace ICA in carefully selected patients.

Coronary Artery Disease↗

"Bounce at the Bell": a novel program of short bouts of exercise improves proximal femur bone mass in early pubertal children.

OBJECTIVES: To examine the effects of a simple and inexpensive physical activity intervention on change in bone mass and structure in school aged children. METHODS: Fifty one children (n = 23 boys and 28 girls; mean age 10.1 years) participated in "Bounce at the Bell" which consisted of 10 counter-movement jumps 3x per day (total approximately 3 min/day). Controls were 71 matched children who followed usual school practice. We assessed dietary calcium, physical activity, physical performance, and anthropometry in September and after 8 months of intervention (June). We measured bone mineral content (BMC) and bone area at the lumbar spine, total body, and proximal femur. Proximal femur scans were also analysed for bone geometry and structural strength using the hip structural analysis program. Lean and fat mass (g) were also calculated. RESULTS: Groups were similar at baseline and did not differ in weight, height, total body, lumbar spine, proximal femur, or femoral neck BMC. Control children had a greater increase in adjusted total body BMC (1.4%). Intervention children gained significantly more BMC at the total proximal femur (2%) and the intertrochanteric region (27%). Change in bone structural parameters did not differ between groups. CONCLUSIONS: This novel, easily implemented exercise program, took only a few minutes each day and enhanced bone mass at the weight bearing proximal femur in early pubertal children. A large, randomised study of boys and girls should be undertaken powered to test the effectiveness of Bounce at the Bell in children at different stages of maturity, and in boys and girls independently.

Analysis of Variance↗

Image quality and diagnostic accuracy of non-invasive coronary imaging with 16 detector slice spiral computed tomography with 188 ms temporal resolution.

OBJECTIVE: To evaluate image quality and clinical accuracy in detecting coronary artery lesions with a new multidetector spiral computed tomography (MDCT) generation with 16 detector slices and a temporal resolution of 188 ms. METHODS: 124 consecutive patients scheduled for invasive coronary angiography (ICA) were additionally studied by MDCT (Sensation 16 Speed 4D). MDCTs were analysed with regard to image quality and presence of coronary artery lesions. The results were compared with ICA. RESULTS: 120 of 124 scans were successful. The image quality of all remaining 120 scans was sufficient (mean (SD) heart rate 64.2 (9.8) beats/min, range 43-95). The mean calcium mass was 167 (223) mg (range 0-1038). Thirteen coronary segments were evaluated for each patient (1560 segments in total). Image quality was graded as follows: excellent, 422 (27.1%) segments; good, 540 (34.6%) segments; moderate, 277 (17.7%) segments; heavily calcified, 215 (13.8%) segments; and blurred, 106 (6.8%) segments. ICA detected 359 lesions with a diameter stenosis > 50% and MDCT detected 304 of 359 (85%). Sensitivity, specificity, and positive and negative predictive values were 85%, 98%, 91%, and 96%, respectively. The correct clinical diagnosis (presence or absence of at least one stenosis > 50%) was obtained for 110 of 120 (92%) patients. CONCLUSIONS: MDCT image quality can be further improved with 16 slices and faster gantry rotation time. These results in an unselected population underline the potential of MDCT to become a non-invasive diagnostic alternative, especially for the exclusion of coronary artery disease, in the near future.

Aged↗

Relationship between Alzheimer's disease clinical stage and Gq/11 in subcellular fractions of frontal cortex.

Alzheimer's disease (AD) is associated with impaired coupling of cell surface muscarinic cholinergic receptors to G proteins of the Gq/11 class in brain. This alteration may contribute to progression of cognitive impairment during the course of the disease. We hypothesized that increasing severity of cognitive impairment would be related to decreased levels of Gq/11 detected in key subcellular fractions made from postmortem brain tissue. In this study, we used Western blotting to determine the quantity of Gq/11alpha in P2, synaptic plasma membrane, cytoplasm, microsomal membrane, and lipid raft fractions prepared from superior frontal cortex gray matter of 25 patients with clinical AD confirmed by post-mortem examination. Multiple linear regression analysis that adjusted for age, sex, and education showed a linear relationship between frontal cortex synaptic plasma membrane Gq/11alpha levels and severity of cognitive impairment determined by Mini Mental State score measured proximate to death.

Age Factors↗

Biochemical and immunological properties of cytokines conjugated to dendritic polymers.

Here we describe a post-translational modification of SC-63032, a variant of the species restricted, multi-lineage hematopoeitic factor human interleukin-3 (hIL-3). We have made two new dendritic polymer (polyamidoamine or PAMAM dendrimers, generation 5)-SC-63032 bioconjugates. Using two distinct chemistries (one of which is novel to this work), we achieved site-specific conjugation with respect to the amino acid in the proteins ligated to the dendrimers. In both bioconjugates, conjugated cytokine maintains its ability to bind the hIL-3 alpha receptor subunit, but is significantly (about 10-fold) less potent in inducing hIL-3 dependent in vitro cell proliferation than is the free cytokine. In vivo data indicates that conjugation decreases the immunogenicity of the conjugated cytokine modestly. In the absence of pharmacokinetic or biodistribution effects associated with the bioconjugates that increase their potency in vivo (which can only be tested in a higher primate, due to the species restriction of hIL-3 and its derivatives), these immune mitigation effects may be too small to be therapeutically significant. Though unmodified PAMAM dendrimers fail to elicit an antibody response in mice, protein conjugation to dendrimers haptenizes them, and a dendrimer-specific antibody response is produced. In toto, the principal limitation of the dendrimer-cytokine bioconjugates herein is in their reduced receptor affinity and potency in vitro. Were the in vivo potency of the bioconjugates to parallel the in vitro potency of the conjugates reported here, it is likely that particular dendrimer bioconjugates could not justify their higher costs of goods relative to the parent SC-63032 molecule, though retention of SC-63032 biological activities in conjugates suggests that other cytokine-dendrimer bioconjugates may be bioactive. This is good news to the nanotechnology community, in as much as PAMAM dendrimers are among the monodisperse polymeric nanomaterials available, and these results show that they can be used successfully in conjugates to bioactive proteins.

Animals↗

Non-invasive evaluation of atherosclerosis with contrast enhanced 16 slice spiral computed tomography: results of ex vivo investigations.

OBJECTIVE: To evaluate the diagnostic accuracy of 16 slice computed tomography (CT) in determining plaque morphology and composition in an experimental setting. The results were compared with histopathological analysis as the reference standard. METHODS: Nine human popliteal arteries derived from amputations because of atherosclerotic disease were investigated with multislice spiral CT (MSCT). Atherosclerotic lesions were morphologically classified (completely or partially occlusive, concentric, eccentric), and tissue densities were determined within these plaques. In addition, vessel dimensions were quantitatively measured. RESULTS: The results were compared with histological analysis. The concordance index kappa for morphological classification was 0.88. Plaque density (n = 51 lesions) was significantly different (p < 0.0001) between lipid rich, fibrotic, and calcified lesions (Stary stage III: n = 2, 58 (8) Hounsfield units (HU); Stary V: n = 11, 50 (21) HU; Stary VI: n = 14, 96 (42) HU; Stary VII: n = 6, 858 (263) HU; Stary VIII: n = 18, 126 (99) HU). The concordance index kappa for the classification of plaques based on density was 0.51. Vessel dimensions had a good correlation (r = 0.98). CONCLUSIONS: 16 slice CT was found to be a reliable non-invasive imaging technique for assessing atherosclerotic plaque morphology and composition. Although calcified lesions can be differentiated from non-calcified lesions, the diagnostic accuracy in further subclassifying non-calcified plaques as lipid rich and fibrotic is low, even under experimental conditions.

Contrast Media↗

[Unilateral apophyseal fracture of the superior anterior iliac crest--a case report].

While soccer playing a 19 year old man suffered from acute onset of severe pain in the anterior part of the iliac crest. A CT-scan confirmed the unilateral avulsion of the anterior superior iliac spine. Due to only minimal fragment dislocation and still open apophysis we decided to treat this injury conservatively with crutches and partial weight bearing. There were no complications and already three weeks after the accident the young man was nearly painfree. Avulsion of the anterior superior iliac spine most commonly occur in adolescents. The fracture is often missed and treated as muscle injury or hip pain and therefore received a symptomatical therapy. Later a typical hypertrophic callus formation confirms the avulsion fracture. The avulsion occurs by sudden pull of the sartorius muscle to the anterior superior iliac spine. The pull of the muscle dislocates the loose fragment caudally. In most of the cases a conservative treatment is successful.

Adult↗

Detection of Mycoplasma pneumoniae by real-time nucleic acid sequence-based amplification.

Real-time isothermal nucleic acid sequence-based amplification (RT-NASBA) was applied to the detection of Mycoplasma pneumoniae. In vitro-generated M. pneumoniae RNA was used to assess the sensitivity of the assay. The 95% hit rate was 148 molecules of M. pneumoniae RNA in the amplification and 10(4) molecules of in vitro-generated RNA after nucleic acid extraction. The sensitivity of the RT-NASBA and the conventional NASBA assays corresponded to 5 color-changing units (CCU) of M. pneumoniae. In spiked throat swabs, nasopharyngeal aspirates, bronchoalveolar lavages, and sputum, the sensitivity of both NASBA assays corresponded to 5 to 50 CCU of M. pneumoniae. A total of 17 clinical specimens positive for M. pneumoniae by PCR were also positive by conventional NASBA, but one specimen was negative by RT-NASBA. These results indicate that the sensitivity of detection of M. pneumoniae by RT-NASBA in respiratory samples might be slightly reduced compared to that by conventional NASBA. However, the real-time assay is superior in speed and ease of handling.

Humans↗

Maladaptive behaviour in children and adolescents with Down's syndrome.

BACKGROUND: Although children with Down's syndrome (DS) are at lower risk for psychopathology than others with intellectual disability, they do show more problems than typically developing children. However, age-related trends in these problems remain unclear. METHODS: The present authors examined age-related changes in the maladaptive behaviours of 211 children and adolescents with DS aged between 4 and 19 years (mean = 9.74 years). Most participants (n = 180) were recruited from families residing in the greater Los Angeles area, California, USA, while a minority (n = 31) were patients from a clinic specializing in the psychiatric management of people with DS. The participants were divided into four age groups: (1) 4-6 years, (2) 7-9 years, (3) 10-13 years and (4) 14-19 years. RESULTS: Externalizing behaviours were lower across both the community and clinic samples, while internalizing behaviours were significantly higher in older adolescents aged between 14 and 19 years. Increases were found in withdrawal, seen in 63% of community-based adolescents, and 75% of clinic adolescents. CONCLUSIONS: Older adolescents with DS may show decreased externalizing symptoms and subtle increases in withdrawal. Possible relationships are discussed between these shifts and increased risks of later-onset depression and Alzheimer's disease in adults with DS.

Adolescent↗

Unstable scaphoid fracture nonunion: a medium-term study of anterior wedge grafting procedures.

A retrospective review of 37 patients with scaphoid fracture nonunions treated by interpositional bone grafting and internal fixation was conducted at an average follow-up of 5.7 years. Solid radiographic union was achieved in 35 cases. Preexisting avascular necrosis was a major adverse factor for achievement of union and satisfactory outcome. Based on the modified Mayo wrist-scoring system, 15 patients had an excellent result, 11 had a good result, four had a fair result and seven had a poor result. Patients with preexisting degenerative changes had a significantly worse clinical outcome. The vast majority of the patients had satisfactory correction of scaphoid length and the associated dorsal intercalated segment instability (DISI). Although 30 patients showed radiographic evidence of mild or moderate degenerative changes at their latest follow-up, there was no significant progression of arthrosis and the scaphoid nonunion advanced carpal collapse deformity did not progress after healing of the fracture nonunion.

Adult↗

Molecular aptamer for real-time oncoprotein platelet-derived growth factor monitoring by fluorescence anisotropy.

Monitoring proteins in real time and in homogeneous solution has always been a difficult task. We have applied a fluorophore-labeled molecular probe based on a high-affinity platelet-derived growth factor (PDGF) aptamer for the ultrasensitive detection of PDGF in homogeneous solutions. The aptamer is labeled with fluorescein to specifically bind with the PDGF protein. Fluorescence anisotropy is used for the real-time monitoring of the binding between the aptamer and the protein. When the labeled aptamer is bound with its target protein, the rotational motion of the fluorophore attached to the complex becomes much slower because of an increased molecular weight after binding, resulting in a significant fluorescence anisotropy change. Using the anisotropy change, we are able to detect the binding events between the aptamer and the protein in real time and in homogeneous solutions (detection without separation). This assay is highly selective and ultrasensitive. It can detect PDGF in the subnanomolar range. The new method for protein detection is simple and inherits all of the advantages of molecular aptamers. Efficient oncoprotein detection using aptamer-based fluorescence anisotropy measurement will find wide applications in protein monitoring, in cancer diagnosis as well as other studies in which protein analysis is important.

Anisotropy↗