Early detection of leukaemic relapse after bone marrow transplantation using the polymerase chain reaction

We report a case of acute lymphoblastic leukaemia relapsing after allogeneic bone marrow transplantation in which the polymerase chain reaction (PCR) was used to assess chimeric status. This technique demonstrated the progressive reappearance of host cells prior to clinical relapse. The relapse was of host cell origin as shown by the presence of female (recipient) metaphases containing an abnormal chromosomal marker (iso 9q) which had also been present at initial diagnosis. The emergence of host cells in this case, detected only by PCR techniques but not by cytogenetic methods, appeared to herald overt relapse. PCR analysis provides a sensitive tool for detecting a progressive rise in host cell numbers which may predict clinical relapse.

Chimaerism following allogeneic bone marrow transplantation:detection of residual host cells using the polymerase chain reaction

Chimaerism was assessed in five recipients following sex mismatched allogeneic bone marrow transplantation. Techniques included karyotyping of bone marrow cells, dot blot DNA analysis of blood and bone marrow suspensions, and in vitro amplification of DNA by the polymerase chain reaction (PCR) using blood and bone marrow suspensions and stored bone marrow slides. Results of karyotypic analysis suggested complete chimaerism in four patients, while in one patient mixed chimaerism was detected. Mixed chimaerism was also detected, however, in a second patient using PCR and confirmed by dot blot analysis on all tissues examined. PCR is a sensitive tool for investigation of chimaerism following bone marrow transplantation. Since this technique does not require radioactivity, it is an attractive method for use in a clinical laboratory. This technique represents a further development in the use of DNA methodologies in the assessment of haematological disease.

A rapid dot-blot assay to assess chimerism following sex-mismatched bone marrow transplantation

Mixed chimerism may occur more frequently than previously thought following allogeneic bone marrow transplantation and may have implications in terms of relapse, graft-versus-host disease and immune reconstitution. DNA analysis using single or multilocus polymorphic probes cannot reliably discriminate between donor and recipient cells below a level of 10%. We used probe pHY2.1, a cloned segment of tandemly repeated DNA (2000 copies) on the long arm of chromosome Y. A dot blot procedure allowed us to immobilize DNA directly from 50 microliter of peripheral blood or bone marrow. Cross-reactivity was eliminated by hybridization at conditions of extreme stringency (65 degrees C, 50% formamide). Mixing experiments detected male DNA at a level of 0.1% after 10 h exposure. Five patients were studied serially post-bone marrow transplantation. One patient showed mixed chimerism for 12 months, one had complete autologous recovery and the remaining three showed complete engraftment. All results were verified by standard karyotyping on bone marrow cells. This technique is a simple, rapid and sensitive assay for chimerism following sex mismatched bone marrow transplantation.

Wing Panel Design with Novel Skin-Buckling Containment Features

The impact of buckling containment features on the stability of thin-gauge fuselage, metallic stiffened panels has previously been demonstrated. With the continuing developments in manufacturing technology, such as welding, extrusion, machining, and additive layer manufacture, understanding the benefits of additional panel design features on heavier applications, such as wing panels, is timely. This compression testing of thick-gauge panels with and without buckling containment features has been undertaken to verify buckling and collapse behaviors and validate sizing methods. The experimental results demonstrated individual panel mass savings on the order of 9%, and wing cover design studies demonstrated mass savings on the order of 4 to 13%, dependent on aircraft size and material choice.

Assessment of the double-skin façade passive thermal buffer effect.

Double Skin Façades (DSFs) are becoming increasingly popular architecture for commercial office buildings. Although DSFs are widely accepted to have the capacity to offer significant passive benefits and enable low energy building performance, there remains a paucity of knowledge with regard to their operation. Identification of the most determinant architectural parameters of DSFs is the focus of ongoing research. This paper presents an experimental and simulation study of a DSF installed on a commercial building in Dublin, Ireland. The DSF is south facing and acts to buffer the building from winter heat losses, but risks enhancing over-heating on sunny days. The façade is extensively monitored during winter months. Computational Fluid Dynamic (CFD) models are used to simulate the convective operation of the DSF. This research concludes DSFs as suited for passive, low energy architecture in temperature climates such as Ireland but identifies issues requiring attention in DSF design.

Experimental analysis of air flow profiles in a double skin facade in a maritime climate

Glazed Double Skin Facades (DSF) offer the potential to improve the performance of all-glass building skins, common to commercial office buildings in which full facade glazing has almost become the standard. Single skin glazing results in increased heating and cooling costs over opaque walls, due to lower thermal resistance of glass, and the increased impact of solar gain through it. However, the performance benefit of DSF technology continues to be questioned and its operation poorly understood, particularly the nature of airflow through the cavity. This paper deals specifically with the experimental analysis of the air flow characteristics in an automated double skin façade. The benefit of the DSF as a thermal buffer, and to limit overheating is evaluated through analysis of an extensive set of parameters including air and surface temperatures at each level in the DSF, airflow readings in the cavity and at the inlet and outlet, solar and wind data, and analytically derived pressure differentials. The temperature and air-flow are monitored in the cavity of a DSF using wireless sensors and hot wire anemometers respectively. Automated louvre operation and building set-points are monitored via the BMS. Thermal stratification and air flow variation during changing weather conditions are shown to effect the performance of the DSF considerably and hence the energy performance of the building. The relative pressure effects due to buoyancy and wind are analysed and quantified. This research aims to developed and validate models of DSFs in the maritime climate, using multi-season data from experimental monitoring. This extensive experimental study provides data for training and validation of models.

High-dose therapy and autologous stem cell transplantation in first relapse for diffuse large B cell lymphoma in the rituximab era:an analysis based on data from the European Blood and Marrow Transplantation Registry

Autologous stem cell transplantation (ASCT) consolidation remains the treatment of choice for patients with relapsed diffuse large B cell lymphoma. The impact of rituximab combined with chemotherapy in either first- or second-line therapy on the ultimate results of ASCT remains to be determined, however. This study was designed to evaluate the benefit of ASCT in patients achieving a second complete remission after salvage chemotherapy by retrospectively comparing the disease-free survival (DFS) after ASCT for each patient with the duration of the first complete remission (CR1). Between 1990 and 2005, a total of 470 patients who had undergone ASCT and reported to the European Blood and Bone Transplantation Registry with Medical Essential Data Form B information were evaluated. Of these 470 patients, 351 (74%) had not received rituximab before ASCT, and 119 (25%) had received rituximab before ASCT. The median duration of CR1 was 11 months. The median time from diagnosis to ASCT was 24 months. The BEAM protocol was the most frequently used conditioning regimen (67%). After ASCT, the 5-year overall survival was 63% (95% confidence interval, 58%-67%) and 5-year DFS was 48% (95% confidence interval, 43%-53%) for the entire patient population. Statistical analysis showed a significant increase in DFS after ASCT compared with duration of CR1 (median, 51 months versus 11 months; P < .001). This difference was also highly significant for patients with previous exposure to rituximab (median, 10 months versus not reached; P < .001) and for patients who had experienced relapse before 1 year (median, 6 months versus 47 months; P < .001). Our data indicate that ASCT can significantly increase DFS compared with the duration of CR1 in relapsed diffuse large B cell lymphoma and can alter the disease course even in patients with high-risk disease previously treated with rituximab.

Phylloseptin-PBa—A Novel Broad-Spectrum Antimicrobial Peptide from the Skin Secretion of the Peruvian Purple-Sided Leaf Frog (Phyllomedusa Baltea) Which Exhibits Cancer Cell Cytotoxicity

Antimicrobial peptides from amphibian skin secretion display remarkable broad-spectrum antimicrobial activity and are thus promising for the discovery of new antibiotics. In this study, we report a novel peptide belonging to the phylloseptin family of antimicrobial peptides, from the skin secretion of the purple-sided leaf frog, Phyllomedusa baltea, which was named Phylloseptin-PBa. Degenerate primers complementary to putative signal peptide sites of frog skin peptide precursor-encoding cDNAs were designed to interrogate a skin secretion-derived cDNA library from this frog. Subsequently, the peptide was isolated and identified using reverse phase HPLC and MS/MS fragmentation. The synthetic replicate was demonstrated to have activity against S. aureus, E. coli and C. albicans at concentrations of 8, 128 and 8 mg/L, respectively. In addition, it exhibited anti-proliferative activity against the human cancer cell lines, H460, PC3 and U251MG, but was less active against a normal human cell line (HMEC). Furthermore, a haemolysis assay was performed to assess mammalian cell cytotoxicity of Phylloseptin-PBa. This peptide contained a large proportion of α-helical domain, which may explain its antimicrobial and anticancer activities.

Data Cleaning and Outlier Removal: Application in Human Skin Detection

An outlier removal based data cleaning technique is proposed to
clean manually pre-segmented human skin data in colour images.
The 3-dimensional colour data is projected onto three 2-dimensional
planes, from which outliers are removed. The cleaned 2 dimensional
data projections are merged to yield a 3D clean RGB data. This data
is finally used to build a look up table and a single Gaussian classifier
for the purpose of human skin detection in colour images.

Survival of patients with skin melanoma in Europe increases further: Results of the EUROCARE-5 study

Background In Europe skin melanoma (SM) survival has increased over time. The aims were to evaluate recent trends and differences between countries and regions of Europe.

Methods Relative survival (RS) estimates and geographical comparisons were based on 241,485 patients aged 15 years and over with a diagnosis of invasive SM in Europe (2000-2007). Survival time trends during 1999-2007 were estimated using the period approach, for 213,101 patients. Age, gender, sub-sites and morphology subgroups were considered. 

Results In European patients, estimated 5-year RS was 83% (95% confidence interval, CI 83-84%). The highest values were found for patients resident in Northern (88%; 87-88%) and Central (88%; 87-88%) Europe, followed by Ireland and United Kingdom (UK) (86%; 85-86%) and Southern Europe (83%; 82-83%). The lowest survival was in Eastern Europe (74%; 74-75%). Within regions the intercountry absolute difference in percentage points of RS varied from 4% (North) to 34% (East). RS decreased markedly with patients' age and was higher in women than men. Differences according to SM morphology and skin sub-sites also emerged. Survival has slightly increased from 1999 to 2007, with a small improvement in Northern and the most pronounced improvement in Eastern Europe. 

Discussion SM survival is high and still increasing in European patients. The gap between Northern and Southern and especially Eastern European countries, although still present, diminished over time. Differences in stage distribution at diagnosis may explain most of the geographical differences. However, part of the improvement in survival may be attributed to overdiagnosis from early diagnosis practices.

A HuGE Review and Meta-Analyses of Genetic Associations in New Onset Diabetes after Kidney Transplantation

PURPOSE: New onset diabetes after transplantation (NODAT) is a serious complication following solid organ transplantation. There is a genetic contribution to NODAT and we have conducted comprehensive meta-analysis of available genetic data in kidney transplant populations.

METHODS: Relevant articles investigating the association between genetic markers and NODAT were identified by searching PubMed, Web of Science and Google Scholar. SNPs described in a minimum of three studies were included for analysis using a random effects model. The association between identified variants and NODAT was calculated at the per-study level to generate overall significance values and effect sizes.

RESULTS: Searching the literature returned 4,147 citations. Within the 36 eligible articles identified, 18 genetic variants from 12 genes were considered for analysis. Of these, three were significantly associated with NODAT by meta-analysis at the 5% level of significance; CDKAL1 rs10946398 p = 0.006 OR = 1.43, 95% CI = 1.11-1.85 (n = 696 individuals), KCNQ1 rs2237892 p = 0.007 OR = 1.43, 95% CI = 1.10-1.86 (n = 1,270 individuals), and TCF7L2 rs7903146 p = 0.01 OR = 1.41, 95% CI = 1.07-1.85 (n = 2,967 individuals).

CONCLUSION: Evaluating cumulative evidence for SNPs associated with NODAT in kidney transplant recipients has revealed three SNPs associated with NODAT. An adequately powered, dense genome-wide association study will provide more information using a carefully defined NODAT phenotype.

SPLI is a Chemoattractant for Oral and Skin Fibroblasts

Objectives: Unlike adult dermal wounds, the oral mucosa demonstrates preferential healing characterized by rapid remodeling and re-epithelialisation, with minimal scar formation. Secretory leukocyte protease inhibitor (SLPI) is an epithelial-derived factor with potential for promoting scarless repair. The aims of this study were to: (i) investigate the directed migratory (chemotaxis) response of oral and skin fibroblasts to various concentrations of SLPI; and (ii) compare migratory speed of the two cell types. Methods: Paired oral and skin fibroblasts were seeded at 2x104 cells in six well plates containing glass coverslips, and cultured in DMEM supplemented with 10% FCS for 48hours. Following a period of serum starvation (18hours in DMEM plus 0.5% BSA), coverslips were incorporated within a Dunn chemotaxis chamber containing DMEM with 0.5% BSA +/- SLPI gradients at 0.5, 1 or 2µM concentrations. Using microscopy, the migratory behaviour of cells was digitally captured every 10mins for 18hours, traced with JCell tracking software and resulting co-ordinates statistically analysed using Mathmatica software. Results: At all concentrations SLPI was a significant chemoattractant (p<0.01) for both cell types. However, skin fibroblasts migrated significantly faster than oral cells at each SLPI concentration, with greatest effect observed at the highest dose (skin: 32.0±0.47µm/hr, oral: 13.6±0.23µm/hr). Conclusion: SLPI is a chemoattractant for both oral and skin fibroblasts, and may play an important role in fibroblast recruitment during wound healing. This work was funded by the R&D Office, N.Ireland.