Biomedical subjects

R Beck

Publications and source records attributed to R Beck.

At least 19 recordsLinked to original sources

PubMed2007

Quantitative protein analysis from formalin-fixed tissues: implications for translational clinical research and nanoscale molecular diagnosis.

Owing to its cross-linking effects, it is currently believed that formalin fixation of routinely processed tissues in the clinic prevents protein extraction and profiling. The aim of our study was to develop a robust, fast, standardized, and easy to use technique for the solubilization of non-degraded, full length, and immunoreactive proteins from formalin-fixed tissues for western blot and protein microarray analysis. Sections of routinely processed formalin-fixed and paraffin-embedded tissues of various origin were analysed. After deparaffination, tissues were manually dissected from the slides and transferred into an optimized protein extraction buffer system. Proteins were solubilized and subsequently analysed by western blot and reverse phase protein microarrays. We succeeded in isolating non-degraded, soluble, and immunoreactive proteins from routinely processed formalin-fixed tissues. We were able to detect membrane, cytoplasmic and nuclear proteins at the expected molecular weight. No differences were found in the protein yield and protein abundances between fresh frozen and formalin-fixed tissues. Using western blots and reverse phase protein microarrays, the receptor tyrosine kinase HER2, an important protein target for antibody based cancer treatment, was reliably measured in formalin-fixed breast cancer biopsy samples when compared with measurement by immunohistochemistry and fluorescence in situ hybridization; remarkably, immunohistochemically equivocal cases (score 2+) can be categorized according to HER2 protein abundance. Our new clinically orientated multiplexed protein measurement system may be generally applicable to determine the relative abundances of known disease-related proteins in small amounts of routinely processed formalin-fixed tissue samples for research and diagnosis. This technique may also be used to identify, characterize, and validate known and new protein markers in a variety of human diseases.

Animals↗

PubMed2006

Differences and similarities of cytomegalovirus and pneumocystis pneumonia in HIV-negative immunocompromised patients thin section CT morphology in the early phase of the disease.

The purpose of this study was to assess CT morphology of pneumocystis pneumonia (PcP) and cytomegalovirus (CMV) pneumonia for specific characteristic features, similarities as well as differences, which might contribute to an early diagnosis and, therefore, influence patient management 58 patients were included, 31 with CMV pneumonia and 27 with PcP. All patients with CMV pneumonia had underlying haematological malignancies (n = 31) mainly treated by haematopoietic cell transplantation (HCT) (n = 26). Patients with PcP had haematological malignancies (n = 17) treated by HCT in 6, solid tumours (n = 5) and corticosteroid therapy (n = 5). Thin section CTs were analysed retrospectively by two radiologists. 18 CT morphological criteria were evaluated for presence or absence. Significance was calculated by chi2 test. Interobserver correlation was tested by kappa-statistic (K) Only 5 of the 18 features were found to have significantly different frequencies in the two entities. Apical distribution (p<0.01), mosaic pattern (p<0.01) and homogeneous structure of ground-glass opacities (GGO) (p<0.05) were found more frequently in PcP (each K: 0.7-0.9), whereas small nodules or unsharp demarcation of GGO and consolidation were typical of CMV pneumonia (p<0.05). Peripheral sparing, consolidation and septal thickening inter alia were found equally in both groups In conclusion analysis of craniocaudal distribution, demarcation and structure of infiltrates may be helpful in prioritizing differential diagnosis of CMV pneumonia or PcP. However, some features thought typical for one or the other entities appear with similar frequency in both groups in HIV-negative patients.

Adult↗

PubMed2006

Measurement of the Gerasimov-Drell-Hearn Integrand for 2H from 200 to 800 MeV.

A measurement of the helicity dependence of the total inclusive photoabsorption cross section on the deuteron was carried out at MAMI (Mainz) in the energy range 200<Egamma<800 MeV. The experiment used a 4pi detection system, a circularly polarized tagged photon beam and a frozen-spin target which provided longitudinally polarized deuterons. The contribution to the Gerasimov-Drell-Hearn sum rule for the deuteron determined from the data is 407+/-20(stat)+/-24(syst) mu b for 200<Egamma<800 MeV.

Journal Article↗

PubMed2006

PET and PET/CT studies of tumor tissue oxygenation.

Hypoxia has been identified as a major adverse prognostic factor for tumor progression and for resistance to anticancer treatment. Various approaches have been evaluated to assess tumor hypoxia in vivo. PET imaging in particular has emerged as a promising non-invasive tool to accurately characterize tumor oxygenation, thus offering the potential to optimize and individualize a therapy for patients suffering from cancer. The current PET tracers and animal models for the study of tissue hypoxia including aspects of small animal PET imaging are reviewed in this paper. Special emphasis is placed on human PET studies assessing tumor hypoxia in patients with different tumor entities, various anticancer therapies (chemotherapy, radiation therapy, hypoxia targeting chemotherapy) as well as studies deriving prognostic factors and outcome data. Methodological advantages underpinning PET/CT in the imaging of hypoxia are discussed. The great promise of PET/CT is its potential as a single imaging modality for whole body staging providing both anatomical and biological information on the tumor as a whole. It allows a more precise estimation of the hypoxic tumor volume as well as comparisons on a voxel-by-voxel basis (parametric mapping). Finally, PET and PET/CT are discussed in the framework of individualized therapies with its special significance for intensity modulated radiation therapy and selective dose escalation in hypoxic tumor regions as well as more systemic approaches such as hypoxia-directed cytotoxins.

Biomarkers, Tumor↗

PubMed2005

Evaluation of biometric methods for measuring the anterior chamber depth in the non-contact mode.

PURPOSE: High-resolution biometry of the anterior ocular segment is now becoming more and more important against a background of refractive surgery and the evaluation of potentially accommodative lens replacement materials. The aim of this study was a systematic investigation of the currently available non-contact methods for measuring the anterior chamber depth (ACD). METHODS: The ACDs of 50 phakic eyes of 27 patients aged between 19 and 59 years were measured with the IOL-Master (Zeiss), the AC-Master (Zeiss), the Pentacam (Oculus) and slit-lamp pachymetry by Jaeger (Haag-Streit). RESULTS: The median anterior chamber depth in the investigated eyes was 3.63 mm for the IOL-Master (minimum 2.88 mm, maximum 4.22 mm), 3.802 mm for the AC-Master (2.816 mm-4.373 mm), 3.915 mm for the Pentacam (minimum 2.994 mm, maximum 4.614 mm) and 3.75 mm for Jaeger (2.887 mm-4.29 mm). With a probability of error of alpha=0.05 there were no significant differences concerning the ACD between the methods of Jaeger and AC-Master, Jaeger and IOL-Master, or Pentacam and AC-Master (Wilcoxon and Wilcox). The intra-individual variability was +/-5.4 microm for AC-Master, +/-12.7 microm for Pentacam, +/-24.5 microm for IOL-Master and +/-41.2 microm for Jaeger. The maximum method-dependent difference in ACD determination was 285 microm. CONCLUSIONS: All the methods allow non-contact biometry, but the results might differ due to measuring principles inherent to the system, experience of the examiner and compliance of the patient. Partial coherence interferometry with the AC-Master offers the advantage of measurement exactly along the optical axis with the highest reproducibility and patient compliance.

Adult↗

PubMed2005

Evaluation of ELISA for detection of Trichinella antibodies in muscle juice samples of naturally infected pigs.

The performance characteristics of an ELISA test for trichinellosis in pigs applied to muscle juice was assessed using 314 samples collected from pigs located in endemic areas of Croatia. Peptic digestion was used as the reference method. The diagnostic accuracy of the two compared dilutions (1:10 and 1:100) was considered to be high because the area under the curve (AUC) index was 0.922 and 0.920 for each dilution, respectively. In this study the two graph-receiver operating characteristic (TG-ROC) analysis was used as a tool for selecting cut-off points. Sensitivity, specificity, likelihood ratios, efficiency and Youden's index were used as indices of test accuracy. The cut-off values that minimize overall misclassification cost under an assumption of 3% prevalence were calculated. Our results indicate that the ELISA applied to muscle juice is a highly accurate test and can be adapted to process a large number of samples.

Animals↗

PubMed2005

Comparison of trichinelloscopy with a digestion method for the detection of Trichinella larvae in muscle tissue from naturally infected pigs with low level infections.

The identification of Trichinella infection in pigs in Croatia has traditionally been done by inspection of individual carcasses. In response to outbreaks of human trichinellosis in the last decade, the Croatian Ministry of Agriculture and Forestry instituted compulsory trichinelloscopic examination of tissue from both commercially and privately slaughtered swine. The purpose of this study was to compare trichinelloscopy and artificial digestion for use in samples containing low numbers of larvae. Each assay was used to test 1,769 field positive samples, 290 of which contained 6 or less larvae per gram of muscle tissue. The sensitivity and specificity of trichinelloscopy with 6 or less l pg was 43.4 and 88%, respectively. kappa-Value as a measure of agreement between trichinelloscopy and artificial digestion was 0.27%. It is noteworthy that a considerable number of the 103 (52%) negative animals on trichinelloscopy contained>or=6l pg which is enough to cause clinical trichinellosis. These findings support other studies that indicate trichinelloscopy is not a method of choice and that it is necessary to implement more sensitive procedures such as artificial digestion.

Animals↗

PubMed2005

Measurement of helicity-dependent photoabsorption cross sections on the neutron from 815 to 1825 MeV.

Helicity-dependent total photoabsorption cross sections on the deuteron have been measured for the first time at ELSA (Bonn) in the photon energy range from 815 to 1825 MeV. Circularly polarized tagged photons impinging on a longitudinally polarized LiD target have been used together with a highly efficient 4pi detector system. The data around 1 GeV are not compatible with predictions from existing multipole analyses. From the measured energy range an experimental contribution to the GDH integral on the neutron of [33.9 +/- 5.5(stat) +/- 4.5(syst)] microb is extracted.

Journal Article↗

PubMed2005

Origin of the anomalous low temperature upturn in the resistivity of the electron-doped cuprate superconductors.

The temperature, doping, and field dependences of the magnetoresistance (MR) in Pr2-xCexCuO4-delta films are reported. We distinguish between orbital MR, found when the magnetic field is applied perpendicular to the ab planes, and the nearly isotropic spin MR. The latter, the major MR effect in the superconducting samples, appears in the region of the doping-temperature phase diagram where drho/dT<0, or an upturn in the resistivity appears. We conclude that the upturn originates from spin scattering processes.

Journal Article↗

PubMed2005

[Three-dimensional ultrasound biomicroscopic examinations for haptic differentiation of potentially accommodative intraocular lenses].

PURPOSE: The principal ability of potentially accommodative IOLs is based on an axial shift of the IOL optics induced by the ciliary body action in interaction with a reversible change in haptic angulation. The aim of this study was to investigate the accommodative performance of this new IOL generation. METHOD: The authors have designed a test device to study IOL performance experimentally. These results were extrapolated to the in vivo situation in patients with accommodative implants. RESULTS: The 3D high-resolution presentation of the anterior segment of the eye allows a separation of the IOL haptic elements. In vitro and in the selected in vivo cases a change in haptic angulation <10 degrees in combination with a maximal IOL shift of 0.5 mm was found. CONCLUSIONS: The simulation model used in combination with 3D ultrasound biomicroscopy provides information about the potential of accommodative IOL designs. Conclusions corresponding to changes in haptic angulation during accommodation can be drawn and applied to the in vivo situation.

Accommodation, Ocular↗

PubMed2005

[Epidemic keratoconjunctivitis. Detecting adenoviruses].

OBJECTIVE: The validity of the 15-min adenovirus assay SAS Adenotest was evaluated compared with virus detection by polymerase chain reaction (PCR) from conjunctival swabs. METHODS: In 75 patients with assumed epidemic keratoconjunctivitis, adenovirus detection from conjunctival swabs was performed by the immunochromatographic assay SAS Adenotest and PCR. RESULTS: In 25 patients adenovirus was detected by PCR, 18 of whom were detected by the SAS Adenotest and 7 of whom were not. No false positive results occurred. Sensitivity was 72% and specificity was 100%. CONCLUSIONS: The results indicate that rapid adenovirus detection with the SAS Adenotest is a useful tool in early epidemic keratoconjunctivitis. An additional PCR should be performed when clinical symptoms persist for 5 days or more.

Adenoviridae↗

PubMed2005

Autoantibodies against bactericidal/permeability-increasing protein (BPI-ANCA) in cystic fibrosis patients treated with azithromycin.

Autoantibodies against bactericidal/permeability-increasing protein (BPI-ANCA) were found in patients with cystic fibrosis (CF). It is speculated that they represent a marker of the chronic endobronchial infection and sustained inflammatory response in CF. Our aim was to evaluate whether azithromycin (AZM), through its antiinflammatory effect, could affect the level of BPI-ANCA in CF patients. Eighteen patients with CF aged 5.5-36.3 years (median 15.1) were enrolled in a randomised, double-blind, placebo-controlled trial of AZM (250 mg twice a week to 10 patients) or placebo (8 patients) for 12 weeks. BPI-ANCA levels were recorded pre- and post-treatment and compared to a group of 18 matched healthy controls. Chi-square analysis, Kruskal-Wallis and Mann-Whitney tests were used to compare between the groups. Pre- and post-treatment values were compared using the Wilcoxon Signed-Ranked test. BPI-ANCA was found in 12 CF patients (67%) and four (22%) healthy subjects (P<0.001). The mean BPI-ANCA level was 3.94+/-6.15 U/ml (mean+/-SD) in healthy subjects and 38.11+/-42.34 U/ml in CF patients (P=0.023). The mean BPI-ANCA level was higher in patients with Pseudomonas aeruginosa compared to those without (64+/-35 U/ml and 25+/-41 U/ml respectively, P=0.032). No change in BPI-ANCA levels occurred in the AZM-treated patients [35 (0-127) U/ml (median (range) and 30 (0-120) U/ml, respectively] or in the placebo group [10 (0-66) U/ml and 13 (0-83) U/ml, respectively]. BPI-ANCA levels are significantly higher in patients with CF compared to healthy controls. BPIANCA levels are higher among patients colonised with P. aeruginosa. Twelve weeks of AZM therapy did not lower the BPI-ANCA level in patients with CF.

Adolescent↗

PubMed2004

Experimental check of the Gerasimov-Drell-Hearn sum rule for 1H.

For the first time we checked the fundamental Gerasimov-Drell-Hearn (GDH) sum rule for the proton experimentally in the photon energy range from 0.2-2.9 GeV with the tagged photon facilities at MAMI (Mainz) and ELSA (Bonn). New data of the doubly polarized total cross section difference are presented in the energy range from 1.6 to 2.9 GeV. The contribution to the GDH integral from 0.2-2.9 GeV yields [254+/-5(stat)+/-12(syst)] microb with negative contributions in the Regge regime at photon energies above 2.1 GeV. This trend supports the validity of the GDH sum rule.

Journal Article↗

PubMed2004

Photoproduction of eta-mesic 3He.

The photoproduction of eta-mesic 3He has been investigated using the TAPS calorimeter at the Mainz Microtron accelerator facility MAMI. The total inclusive cross section for the reaction gamma3He-->etaX has been measured for photon energies from threshold to 820 MeV. The total and angular differential coherent eta cross sections have been extracted up to energies of 745 MeV. A resonancelike structure just above the eta production threshold with an isotropic angular distribution suggests the existence of a resonant quasibound state. This is supported by studies of a competing decay channel of such a quasibound eta-mesic nucleus into pi(0)pX. A binding energy of (-4.4+/-4.2) MeV and a width of (25.6+/-6.1) MeV is deduced for the quasibound eta-mesic state in 3He.

Journal Article↗

PubMed2004

Preimplantation genetic diagnosis for early-onset torsion dystonia.

Early-onset primary torsion dystonia (DYT1) is the most severe and common form of hereditary movement disorders, characterized by sustained twisting contractures that begin in childhood, which is caused in majority of cases by a 3-bp deletion of the DYT1 gene on chromosome 9q34 at the heterozygote state. As there is no effective treatment of this disease, preimplantation genetic diagnosis (PGD) may be a useful option for at-risk couples to establish an DYT1 mutation-free pregnancy. PGD was performed for two obligate carriers of the DYT1 3-bp deletion, using blastomere testing to preselect the mutation-free embryos, based on mutation analysis with simultaneous testing of the three closely linked markers, D9S62, D9S63 and ASS. Of 19 tested blastomeres in three cycles, 17 had conclusive information about the mutation and linked markers, of which eight were predicted to be free of 3-bp deletion. Six of these embryos were transferred back to patients, two in each cycle, yielding singleton DYT1 3-bp deletion-free clinical pregnancies in two. One of these pregnancies was terminated due to severe anencephaly and the other resulted in birth of a mutation-free child. This is the first PGD for primary torsion dystonia, providing an alternative for those at-risk couples who cannot accept prenatal diagnosis and termination of pregnancy as an option for avoiding early onset torsion dystonia.

Dystonia Musculorum Deformans↗

PubMed2004

[Orbital manifestation of microscopic polyangiitis (mPA)--presentation of clinical cases].

BACKGROUND: Microscopic polyangiitis (mPA) is one of the ANCA-associated primary systemic vasculitides. It is defined as necrotic vasculitis without granuloma development and without minimal immuno-depots in situ. The clinical picture is characterised by destruction of the small vessels with the main manifestations in the upper and lower respiratory tract, in the lungs and kidneys. The eye manifestations were noted yet not so often. PATIENT: There were 2 clinical cases, aged 5-12 years, with primary eye manifestations of the mPA. The initial oedema and the reddening of the lid area were common to all the patients. The diagnostic excisions of the inflammatory tumour of the anterior orbital area have shown histologically vasculitis of the small vessels. mPA was diagnosed by clinical and immunological parameters, including the examination of the respiratory tract and kidneys. CONCLUSIONS: In inflammatory orbital pseudotumour, diagnostic excision and evaluation of the immunological parameters are useful for the exclusion of a systemic vasculitis. Proper examinations must be performed to determine other possible organ manifestations. Prognosis is characterised by the severity of kidney involvement. Early immunosuppressive therapy is crucially important for survival prognosis.

Child↗

PubMed2004

Influence of methods for Trichinella detection in pigs from endemic and non-endemic European region.

A total of 1401 German and 226 Croatian pigs raised either indoors or outdoors were tested for Trichinella infection by direct and indirect detection methods. A 10 g sample of diaphragm were examined for muscle larvae by the artificial digestion method; the species was determined by multiplex polymerase chain reaction (PCR). For detection of anti-Trichinella IgG, serum samples diluted 1:100, and meat juice samples diluted 1:10, were tested by enzyme-linked immunosorbent assay. All German pigs and those Croatian pigs raised indoors proved to be Trichinella-negative by all methods. Muscle larvae were detected in a total of eleven of the Croatian pigs, which were raised on small outdoor farms. For eight isolates, PCR results demonstrated that recovered larvae were Trichinella spiralis. Anti-Trichinella-IgG was detected in serum and meat juice of digestion positive animals when the worm burdens exceeded 0.38 larvae per gram of muscle. Positive results in Croatian pigs indicate a higher risk of infection for outdoor farming in areas where Trichinella is endemic. Results of direct and indirect detection were compared and are discussed with special regard to specificity and sensitivity of methods.

Abattoirs↗

PubMed2003

First Measurement of the Gerasimov-Drell-Hearn Sum Rule for 1H from 0.7 to 1.8 GeV at ELSA.

To verify the fundamental Gerasimov-Drell-Hearn (GDH) sum rule for the first time experimentally, we measured the helicity dependent total photoabsorption cross section with circularly polarized real photons and longitudinally polarized nucleons in the photon energy range 0.68-1.82 GeV with the tagged photon facility at ELSA. The experiment was carried out with a 4pi detection system, a circularly polarized tagged photon beam, and a frozen spin polarized proton target. The contribution to the GDH sum rule in this photon energy range is [49.9+/-2.4(stat)+/-2.2(syst)] microb.

Journal Article↗