Field camera versus phantom-based measurement of the gradient system transfer function (GSTF) with dwell time compensation.

PURPOSE: The gradient system transfer function (GSTF) can be used to describe the dynamic gradient system and applied for trajectory correction in non-Cartesian MRI. This study compares the field camera and the phantom-based methods to measure the GSTF and implements a compensation for the difference in measurement dwell time. METHODS: The self-term GSTFs of a MR system were determined with two approaches: 1) using a dynamic field camera and 2) using a spherical phantom-based measurement with standard MR hardware. The phantom-based GSTF was convolved with a box function to compensate for the dwell time dependence of the measurement. The field camera and phantom-based GSTFs were used for trajectory prediction during retrospective image reconstruction of 3D wave-CAIPI phantom images. RESULTS: Differences in the GSTF magnitude response were observed between the two measurement methods. For the wave-CAIPI sequence, this led to deviations in the GSTF predicted trajectories of 4% compared to measured trajectories, and residual distortions in the reconstructed phantom images generated with the phantom-based GSTF. Following dwell-time compensation, deviations in the GSTF magnitudes, GSTF-predicted trajectories, and resulting image artifacts were eliminated (< 0.5% deviation in trajectories). CONCLUSION: With dwell time compensation, both the field camera and the phantom-based GSTF self-terms show negligible deviations and lead to strong artifact reduction when they are used for trajectory correction in image reconstruction.

[Pretend play ability in pre-school children]. / Symbolspielkompetenz bei Vorschulkindern.

Children with specific language impairment (SLI) are confronted with limitations in their language abilities that cannot be attributed to cognition, hearing impairments, or neurological deficits. However, there is evidence that children with SLI also have impairments. These include, for example, an impaired ability to pretend play. The current article aims to present firstly normal development of play behavior in children, followed by the Child-Initiated Pretend Play Assessment (ChIPPA). This test enables an objective and standardized assessment of whether a child's ability to initiate and sustain pretend play is age-appropriate. SLI children with impaired play behavior should receive structured individual therapy.

CD87-positive tumor cells in bone marrow aspirates identified by confocal laser scanning fluorescence microscopy.

Dissemination of single tumor cells to the bone marrow is a common event in cancer. The clinical significance of cytokeratin-positive cells detected in the bone marrow of cancer patients is still a matter of debate. In gastric cancer, overexpression of the receptor (uPAR or CD87) for the serine protease urokinase-type plasminogen activator (uPA) in disseminated cancer cells indicates shorter survival of cancer patients. A new immunofluorescence approach, applying confocal laser scanning microscopy, is introduced to locate CD87 antigen in cytokeratin-positive tumor cells and to quantify the CD87 antigen by consecutive scanning. At first, cytokeratin 8/18/19-positive carcinoma cells are identified at excitation wavelength 488 nm using monoclonal antibody A45B/B3 to the cytokeratins and goat anti-mouse IgG labeled with the fluorochrome Alexa488. Next, CD87 in tumor cells is identified by chicken antibody HU277 to the uPA-receptor and goat anti-chicken IgY labeled with fluorochrome Alexa568 (excitation wavelength 568 nm) and the fluorescence signal quantified on a single cell basis using fluorescently labeled latex beads as the fluorescence reference. From 16 patients with gastric or esophageal carcinoma, bone marrow aspirates were obtained, stained for cytokeratins and CD87 and then subjected to laser scanning fluorescence microscopy. Three of six gastric cancer patients had tumor cells present in the bone marrow of which 2 stained for CD87. Three of ten esophageal carcinoma patients had tumor cells in the bone marrow, all three samples stained for CD87. CD87-positive tumor cells were also dissected from stained bone marrow aspirates by laser microdissection microscope to allow analysis of single cells at the gene level.

Epidemiology and clinical features of vivax malaria imported to Europe: sentinel surveillance data from TropNetEurop.

BACKGROUND: Plasmodium vivax is the second most common species among malaria patients diagnosed in Europe, but epidemiological and clinical data on imported P. vivax malaria are limited. The TropNetEurop surveillance network has monitored the importation of vivax malaria into Europe since 1999. OBJECTIVES: To present epidemiological and clinical data on imported P. vivax malaria collected at European level. MATERIAL AND METHODS: Data of primary cases of P. vivax malaria reported between January 1999 and September 2003 were analysed, focusing on disease frequency, patient characteristics, place of infection, course of disease, treatment and differences between network-member countries. RESULTS: Within the surveillance period 4,801 cases of imported malaria were reported. 618 (12.9%) were attributed to P. vivax. European travellers and immigrants were the largest patient groups, but their proportion varied among the reporting countries. The main regions of infection in descending order were the Indian subcontinent, Indonesia, South America and Western and Eastern Africa, as a group accounting for more than 60% of the cases. Regular use of malaria chemoprophylaxis was reported by 118 patients. With 86 (inter-quartile range 41-158) versus 31 days (inter-quartile range 4-133) the median symptom onset was significantly delayed in patients with chemoprophylaxis (p < 0.0001). Common complaints were fever, headache, fatigue, and musculo-skeletal symptoms. All patients survived and severe clinical complications were rare. Hospitalization was provided for 60% and primaquine treatment administered to 83.8% of the patients, but frequencies varied strongly among reporting countries. CONCLUSIONS: TropNetEurop data can contribute to the harmonization of European treatment policies.

Noncontact laser microdissection and catapulting for pure sample capture.

The understanding of the molecular mechanisms of cellular function, growth, and proliferation is based on the accurate identification, isolation, and finally characterization of a specific single cell or a population of cells and its subsets of biomolecules. For the simultaneous analysis of thousands of molecular parameters within one single experiment as realized by DNA, RNA, and protein microarray technologies, a defined number of homogeneous cells derived from a distinct morphological origin are required. Sample preparation is therefore a very crucial step preceding the functional characterization of specific cell populations. Laser microdissection and laser pressure catapulting (LMPC) enables pure and homogeneous sample preparation resulting in an increased specificity of molecular analyses. With LMPC, the force of focused laser light is utilized to excise selected cells or large tissue areas from object slides down to individual single cells and subcellular components like organelles or chromosomes. After microdissection, the sample is directly catapulted into an appropriate collection vial. As this process works entirely without mechanical contact, it enables pure sample retrieval from morphologically defined origin without cross-contamination. LMPC has been successfully applied to isolate and catapult cells from, for example, histological tissue sections, from forensic evidence material, and also from tough plant matter, supporting biomedical research, forensic science, and plant physiology studies. Even delicate living cells like stem cells have been captured for recultivation without affecting their viability or stem cell character, an important feature influencing stem cell research, regenerative medicine, and drug development. The combination of LMPC with microinjection to inject drugs or genetic material into individual cells and to capture them for molecular analyses bears great potential for efficient patient-tailored medication.

Imported leishmaniasis in Germany 2001-2004: data of the SIMPID surveillance network.

Leishmaniasis is a rare, non-notifiable disease in Germany. Epidemiological and clinical data, therefore, are scarce. Most infections seen in Germany are contracted outside the country. The German surveillance network for imported infectious diseases (Surveillance Importierter Infektionen in Deutschland, or SIPMID) recorded 42 cases of imported leishmaniasis (16 visceral, 23 cutaneous, and 3 mucocutaneous) from January 2001 to June 2004. Although most infections were acquired in European Mediterranean countries, the risk of infection was highest for travelers to Latin America. HIV coinfection was observed significantly more often in patients with visceral leishmaniasis than in patients with cutaneous/mucocutaneous leishmaniasis (31 vs. 4%, p=0.02). The median time to a definitive diagnosis was 85 days in cases of visceral leishmaniasis and 61 days in cases of cutaneous/mucocutaneous leishmaniasis, reflecting the unfamiliarity of German physicians with leishmanial infections. Visceral leishmaniasis was treated most frequently with amphotericin B, whereas cutaneous/mucocutaneous leishmaniasis was treated with a variety of local and systemic therapies. The findings presented here should serve to increase awareness as well as improve clinical management of leishmaniasis in Germany.

Self-organized stable pacemakers near the onset of birhythmicity.

General amplitude equations are derived for reaction-diffusion systems near the soft onset of birhythmicity described by a supercritical pitchfork-Hopf bifurcation. Using these equations and applying singular perturbation theory, we show that stable autonomous pacemakers represent a generic kind of spatiotemporal patterns in such systems. This is verified by numerical simulations, which also show the existence of breathing and swinging pacemaker solutions. The drift of self-organized pacemakers in media with spatial parameter gradients is analytically and numerically investigated.

Diagnosis of papillary thyroid carcinoma is facilitated by using an RT-PCR approach on laser-microdissected archival material to detect RET oncogene activation.

OBJECTIVE: The purpose of this study was to investigate the value of the expression of the RET oncogene (rearranged during transfection) in papillary thyroid carcinomas (PTC) and its variants in the differential diagnosis of thyroid neoplasias. According to the literature RET oncogene activation by chromosomal rearrangements has been exclusively implicated in PTCs. METHODS: To establish the incidence of RET activation in PTCs we used 5- to 10-microm sections from archival paraffin blocks. Either parts of the tissue slices were manually dissected or a few distinct cells were microdissected by laser-mediated manipulation with the Robot-MicroBeam system. RNA was extracted from paraffin-embedded thyroid tumors and the corresponding normal tissue. RT and nested PCR were performed using primers for RET/PTC1, PTC2 and PTC3, or for RET exons 12 and 13. PCR products were resolved by gel electrophoresis. RESULTS: We detected RET transcription in approximately 85% of the PTCs including follicular variants and in isolated cells of the same tissues, but not in nonmalignant thyroid tissue. CONCLUSIONS: Our method may serve as an additional diagnostic tool to characterize ambiguous neoplasias and to identify especially nonpapillary, i.e. follicular tumors, as papillary carcinomas. Additionally, this study has demonstrated that expressed genes can be analyzed from routine histopathological tissue slides or pooled single cells. Large retrospective studies can also be performed with this method.