Good manufacturing practice-compliant validation and preparation of BM cells for the therapy of acute myocardial infarction

BACKGROUND: Intracoronary application of BM-derived cells for the treatment of acute myocardial infarction (AMI) is currently being studied intensively. Simultaneously, strict legal requirements surround the production of cells for clinical studies. Thus good manufacturing practice (GMP)-compliant collection and preparation of BM for patients with AMI was established by the Cytonet group. METHODS: As well as fulfillment of standard GMP requirements, including a manufacturing license, validation of the preparation process and the final product was performed. Whole blood (n=6) and BM (n=3) validation samples were processed under GMP conditions by gelafundin or hydroxyethylstarch sedimentation in order to reduce erythrocytes/platelets and volume and to achieve specifications defined in advance. Special attention was paid to the free potassium (<6 mmol/L), some rheologically relevant cellular characteristics (hematocrit <0.45, platelets <450 x 10(6)/mL) and the sterility of the final product. RESULTS: The data were reviewed and GMP compliance was confirmed by the German authorities (Paul-Ehrlich Institute). Forty-five BM cell preparations for clinical use were carried out following the validated methodology and standards. Additionally three selections of CD34+ BM cells for infusion were performed. All specification limits were met. Discussion In conclusion, preparation of BM cells for intracoronary application is feasible under GMP conditions. As the results of sterility testing may not be available at the time of intracoronary application, the highest possible standards to avoid bacterial and other contaminations have to be applied. The increased expense of the GMP-compliant process can be justified by higher safety for patients and better control of the final product.

Haptics in forensics: the possibilities and advantages in using the haptic device for reconstruction approaches in forensic science

Non-invasive documentation methods such as surface scanning and radiological imaging are gaining in importance in the forensic field. These three-dimensional technologies provide digital 3D data, which are processed and handled in the computer. However, the sense of touch gets lost using the virtual approach. The haptic device enables the use of the sense of touch to handle and feel digital 3D data. The multifunctional application of a haptic device for forensic approaches is evaluated and illustrated in three different cases: the representation of bone fractures of the lower extremities, by traffic accidents, in a non-invasive manner; the comparison of bone injuries with the presumed injury-inflicting instrument; and in a gunshot case, the identification of the gun by the muzzle imprint, and the reconstruction of the holding position of the gun. The 3D models of the bones are generated from the Computed Tomography (CT) images. The 3D models of the exterior injuries, the injury-inflicting tools and the bone injuries, where a higher resolution is necessary, are created by the optical surface scan. The haptic device is used in combination with the software FreeForm Modelling Plus for touching the surface of the 3D models to feel the minute injuries and the surface of tools, to reposition displaced bone parts and to compare an injury-causing instrument with an injury. The repositioning of 3D models in a reconstruction is easier, faster and more precisely executed by means of using the sense of touch and with the user-friendly movement in the 3D space. For representation purposes, the fracture lines of bones are coloured. This work demonstrates that the haptic device is a suitable and efficient application in forensic science. The haptic device offers a new way in the handling of digital data in the virtual 3D space.

Non-classical karyotypic features in relapsed childhood B-cell precursor acute lymphoblastic leukemia

Karyotype analysis of acute lymphoblastic leukemia (ALL) at diagnosis has provided valuable prognostic markers for treatment stratification. However, reports of cytogenetic studies of relapsed ALL samples are limited. We compared the karyotypes from 436 nonselected B-cell precursor ALL patients at initial diagnosis and of 76 patients at first relapse. We noticed a relative increase of karyotypes that did not fall into the classic ALL cytogenetic subgroups (high hyperdiploidy, t(12;21), t(9;22), 11q23, t(1;19), <45 chromosomes) in a group of 29 patients at relapse (38%) compared to 130 patients at presentation (30%). Non-classical cytogenetic aberrations in these 29 patients were mostly found on chromosomes 1, 2, 7, 9, 13, 14, and 17. We also describe six rare reciprocal translocations, three of which involved 14q32. The most frequent abnormalities were found in 9p (12/29 cases) and were associated with a marked decrease in the duration of the second remission, but not of the probability of 10-year event-free survival after relapse treatment. From 29 patients with non-classical cytogenetic aberrations, only 8 (28%) had been stratified to a high risk-arm on the first treatment protocol, suggesting that this subgroup might benefit from the identification of new prognostic markers in future studies.

Statistical analysis of personal radiofrequency electromagnetic field measurements with nondetects

Exposimeters are increasingly applied in bioelectromagnetic research to determine personal radiofrequency electromagnetic field (RF-EMF) exposure. The main advantages of exposimeter measurements are their convenient handling for study participants and the large amount of personal exposure data, which can be obtained for several RF-EMF sources. However, the large proportion of measurements below the detection limit is a challenge for data analysis. With the robust ROS (regression on order statistics) method, summary statistics can be calculated by fitting an assumed distribution to the observed data. We used a preliminary sample of 109 weekly exposimeter measurements from the QUALIFEX study to compare summary statistics computed by robust ROS with a naïve approach, where values below the detection limit were replaced by the value of the detection limit. For the total RF-EMF exposure, differences between the naïve approach and the robust ROS were moderate for the 90th percentile and the arithmetic mean. However, exposure contributions from minor RF-EMF sources were considerably overestimated with the naïve approach. This results in an underestimation of the exposure range in the population, which may bias the evaluation of potential exposure-response associations. We conclude from our analyses that summary statistics of exposimeter data calculated by robust ROS are more reliable and more informative than estimates based on a naïve approach. Nevertheless, estimates of source-specific medians or even lower percentiles depend on the assumed data distribution and should be considered with caution.

Patterned-ground plant communities along a bioclimate gradient in the High Arctic, Canada

Non-sorted circles, non-sorted polygons, and earth hummocks are common ground-surface features ill arctic regions. The), are caused by a variety of physical processes that Occur in permafrost regions including contraction cracking and frost heave. Here we describe the vegetation of patterned-ground forms on zonal sites at three location!: along an N-S transect through the High Arctic of Canada. We made 75 releves on patterned-ground features (circles, polygons, earth hummocks) and adjacent tundra (Interpolygon, intercircle, interhummock areas) and identified and classified the vegetation according to the Braun-Blanquet Method. Environmental factors were correlated with the vegetation data using a nonmetric multidimensional scaling ordination (NMDS). We identified eleven commnunities: (1) Puccinellia angustata-Papaver radicalum community in xeromesic non-sorted polygons of subzone A of the Circumpolar Arctic Vegetation Map; (2) Saxifraga-Parmelia omphalodes ssp. glacialis community in hydromesic interpolygon areas of subzone A; (3) Hypogymnia subobscura-Lecanora epibryon community In xeromesic non-sorted polygons of subzone B; (4) Orthotrichum speciosum-Salix arctica community In xeromesic interpolygon areas of subzone B; (5) Cochlearia groenlandica-Luzula nivalis community in hydromesic earth Mocks Of subzone B; (6) Salix arctica-Eriophorum angustifolium ssp. triste community in hygric earth hummocks of subzone 13; (7) Puccinellia angustata-Potentilla vahliana community in xeromesic non-sorted circles and bare patches of subzone Q (8) Dryas integrifolia-Carex rupestris community in xeromesic intercircle areas and vegetated patches of subzone C; (9) Braya glabella ssp. purpurascens-Dryas integrifolia community In hydromesic non-sorted circles of subzone Q (10) Dryas integrifolia-Carex aquatilis community in hydromesic intercircle areas of subzone C; and (11) Eriophorum angustifolium ssp. triste-Carex aquatilis community ill hygric intercircle areas of subzone C. The NMDS ordination displayed the vegetation types with respect to complex environmental gradients. The first axis of the ordination corresponds to a complex soil moisture gradient and the second axis corresponds to a complex geology/elevation/climate gradient. The tundra plots have a greater moss and graminoid cover than the adjacent frost-heave communities. In general, frost-heave features have greater thaw depths, more bare ground, thinner organic horizons, and lower soil moisture than the surrounding tundra. The morphology of the investigated patterned ground forms changes along the climatic gradient, with non-sorted pollygons dominating in the northernmost sites and non-sorted circles dominating, in the southern sites.