Establishing optimal quantitative-polymerase chain reaction assays for routine diagnosis and tracking of minimal residual disease in JAK2-V617F-associated myeloproliferative neoplasms:a joint European LeukemiaNet/MPN&MPNr-EuroNet (COST action BM0902) study

Reliable detection of JAK2-V617F is critical for accurate diagnosis of myeloproliferative neoplasms (MPNs); in addition, sensitive mutation-specific assays can be applied to monitor disease response. However, there has been no consistent approach to JAK2-V617F detection, with assays varying markedly in performance, affecting clinical utility. Therefore, we established a network of 12 laboratories from seven countries to systematically evaluate nine different DNA-based quantitative PCR (qPCR) assays, including those in widespread clinical use. Seven quality control rounds involving over 21,500 qPCR reactions were undertaken using centrally distributed cell line dilutions and plasmid controls. The two best-performing assays were tested on normal blood samples (n=100) to evaluate assay specificity, followed by analysis of serial samples from 28 patients transplanted for JAK2-V617F-positive disease. The most sensitive assay, which performed consistently across a range of qPCR platforms, predicted outcome following transplant, with the mutant allele detected a median of 22 weeks (range 6-85 weeks) before relapse. Four of seven patients achieved molecular remission following donor lymphocyte infusion, indicative of a graft vs MPN effect. This study has established a robust, reliable assay for sensitive JAK2-V617F detection, suitable for assessing response in clinical trials, predicting outcome and guiding management of patients undergoing allogeneic transplant.

On the MIMO Capacity with Residual Transceiver Hardware Impairments

Radio-frequency (RF) impairments in the transceiver hardware of communication systems (e.g., phase noise (PN), high power amplifier (HPA) nonlinearities, or in-phase/quadrature-phase (I/Q) imbalance) can severely degrade the performance of traditional multiple-input multiple-output (MIMO) systems. Although calibration algorithms can partially compensate these impairments, the remaining distortion still has substantial impact. Despite this, most prior works have not analyzed this type of distortion. In this paper, we investigate the impact of residual transceiver hardware impairments on the MIMO system performance. In particular, we consider a transceiver impairment model, which has been experimentally validated, and derive analytical ergodic capacity expressions for both exact and high signal-to-noise ratios (SNRs). We demonstrate that the capacity saturates in the high-SNR regime, thereby creating a finite capacity ceiling. We also present a linear approximation for the ergodic capacity in the low-SNR regime, and show that impairments have only a second-order impact on the capacity. Furthermore, we analyze the effect of transceiver impairments on large-scale MIMO systems; interestingly, we prove that if one increases the number of antennas at one side only, the capacity behaves similar to the finite-dimensional case. On the contrary, if the number of antennas on both sides increases with a fixed ratio, the capacity ceiling vanishes; thus, impairments cause only a bounded offset in the capacity compared to the ideal transceiver hardware case.

Impact of Residual Transmit RF Impairments on Training-Based MIMO Systems

Radio-frequency (RF) impairments, which intimately exist in wireless communication systems, can severely limit the performance of multiple-input-multiple-output (MIMO) systems. Although we can resort to compensation schemes to mitigate some of these impairments, a certain amount of residual impairments always persists. In this paper, we consider a training-based point-to-point MIMO system with residual transmit RF impairments (RTRI) using spatial multiplexing transmission. Specifically, we derive a new linear channel estimator for the proposed model, and show that RTRI create an estimation error floor in the high signal-to-noise ratio (SNR) regime. Moreover, we derive closed-form expressions for the signal-to-noise-plus-interference ratio (SINR) distributions, along with analytical expressions for the ergodic achievable rates of zero-forcing, maximum ratio combining, and minimum mean-squared error receivers, respectively. In addition, we optimize the ergodic achievable rates with respect to the training sequence length and demonstrate that finite dimensional systems with RTRI generally require more training at high SNRs than those with ideal hardware. Finally, we extend our analysis to large-scale MIMO configurations, and derive deterministic equivalents of the ergodic achievable rates. It is shown that, by deploying large receive antenna arrays, the extra training requirements due to RTRI can be eliminated. In fact, with a sufficiently large number of receive antennas, systems with RTRI may even need less training than systems with ideal hardware.

Minimal residual disease detection in childhood acute lymphoblastic leukaemia patients at multiple time-points reveals high levels of concordance between molecular and immunophenotypic approaches

In this single centre study of childhood acute lymphoblastic leukaemia (ALL) patients treated on the Medical Research Council UKALL 97/99 protocols, it was determined that minimal residual disease (MRD) detected by real time quantitative polymerase chain reaction (RQ-PCR) and 3-colour flow cytometry (FC) displayed high levels of qualitative concordance when evaluated at multiple time-points during treatment (93.38%), and a combined use of both approaches allowed a multi time-point evaluation of MRD kinetics for 90% (53/59) of the initial cohort. At diagnosis, MRD markers with sensitivity of at least 0.01% were identified by RQ-PCR detection of fusion gene transcripts, IGH/TRG rearrangements, and FC. Using a combined RQ-PCR and FC approach, the evaluation of 367 follow-up BM samples revealed that the detection of MRD >1% at Day 15 (P = 0.04), >0.01% at the end of induction (P = 0.02), >0.01% at the end of consolidation (P = 0.01), >0.01% prior to the first delayed intensification (P = 0.01), and >0.1% prior to the second delayed intensification and continued maintenance (P = 0.001) were all associated with relapse and, based on early time-points (end of induction and consolidation) a significant log-rank trend (P = 0.0091) was noted between survival curves for patients stratified into high, intermediate and low-risk MRD groups.

Wilms' tumor gene 1 (WT1) expression in childhood acute lymphoblastic leukemia:a wide range of WT1 expression levels, its impact on prognosis and minimal residual disease monitoring

Wilms' tumor gene 1 (WT1) is overexpressed in the majority (70-90%) of acute leukemias and has been identified as an independent adverse prognostic factor, a convenient minimal residual disease (MRD) marker and potential therapeutic target in acute leukemia. We examined WT1 expression patterns in childhood acute lymphoblastic leukemia (ALL), where its clinical implication remains unclear. Using a real-time quantitative PCR designed according to Europe Against Cancer Program recommendations, we evaluated WT1 expression in 125 consecutively enrolled patients with childhood ALL (106 BCP-ALL, 19 T-ALL) and compared it with physiologic WT1 expression in normal and regenerating bone marrow (BM). In childhood B-cell precursor (BCP)-ALL, we detected a wide range of WT1 levels (5 logs) with a median WT1 expression close to that of normal BM. WT1 expression in childhood T-ALL was significantly higher than in BCP-ALL (P<0.001). Patients with MLL-AF4 translocation showed high WT1 overexpression (P<0.01) compared to patients with other or no chromosomal aberrations. Older children (> or =10 years) expressed higher WT1 levels than children under 10 years of age (P<0.001), while there was no difference in WT1 expression in patients with peripheral blood leukocyte count (WBC) > or =50 x 10(9)/l and lower. Analysis of relapsed cases (14/125) indicated that an abnormal increase or decrease in WT1 expression was associated with a significantly increased risk of relapse (P=0.0006), and this prognostic impact of WT1 was independent of other main risk factors (P=0.0012). In summary, our study suggests that WT1 expression in childhood ALL is very variable and much lower than in AML or adult ALL. WT1, thus, will not be a useful marker for MRD detection in childhood ALL, however, it does represent a potential independent risk factor in childhood ALL. Interestingly, a proportion of childhood ALL patients express WT1 at levels below the normal physiological BM WT1 expression, and this reduced WT1 expression appears to be associated with a higher risk of 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.

Predicting impact damage, residual strength and crashworthiness of composite structures

The development of the latest generation of wide-body carbon-fibre composite passenger aircraft has heralded a new era in the utilisation of these materials. The premise of superior specific strength and stiffness, corrosion and fatigue resistance, is tempered by high development costs, slow production rates and lengthy and expensive certification programmes. Substantial effort is currently being directed towards the development of new modelling and simulation tools, at all levels of the development cycle, to mitigate these shortcomings. One of the primary challenges is to reduce the extent of physical testing, in the certification process, by adopting a ‘certification by simulation’ approach. In essence, this aspirational objective requires the ability to reliably predict the evolution and progression of damage in composites. The aerospace industry has been at the forefront of developing advanced composites modelling tools. As the automotive industry transitions towards the increased use of composites in mass-produced vehicles, similar challenges in the modelling of composites will need to be addressed, particularly in the reliable prediction of crashworthiness. While thermoset composites have dominated the aerospace industry, thermoplastics composites are likely to emerge as the preferred solution for meeting the high-volume production demands of passenger road vehicles. This keynote presentation will outline recent progress and current challenges in the development of finite-element-based predictive modelling tools for capturing impact damage, residual strength and energy absorption capacity of thermoset and thermoplastic composites for crashworthiness assessments.

Alguns aspectos da modelação ecológica na costa portuguesa (Aveiro) : efeitos físicos na distribuição de nutrientes e biomassa fitoplanctónica

O presente trabalho propõe estudar alguns aspectos da modelação ecológica na Costa Portuguesa, nomeadamente o efeito físico na distribuição de nutrientes e biomassa fitoplanctónica. O principal propósito foi implementar e validar um modelo acoplado tridimensional físico e ecológico, para a costa portuguesa, e aplicá-lo numa área limitada, a norte, por Vila do Conde e a sul pela Figueira da Foz centrada na região de Aveiro, para estudar a distribuição, vertical e horizontal, de temperatura, nutrientes e biomassa fitoplanctónica. A região em estudo está situada na costa oeste da Península Ibérica e faz parte da Região de Afloramento do Atlântico Norte. É caracterizada por condições meteorológicas onde a predominância e prevalência, durante uma grande parte do ano, de ventos de norte/noroeste, constitui um dos principais elementos forçadores do transporte para o largo das águas costeiras e consequente subida das águas mais frias e profundas, ricas em nutrientes. A estas condições juntam-se as boas condições de luminosidade e temperatura essenciais ao desenvolvimento fitoplanctónico, que servirá de alimento às espécies marinhas. Este facto, torna esta região, uma zona de elevada riqueza biológica favorável ao desenvolvimento de várias espécies marinhas, transformando este local num ecossistema de forte produtividade. O modelo foi calibrado e validado para a área em estudo e simulou com sucesso, a resposta do sistema à situação de ventos favoráveis ao afloramento costeiro para a região em estudo. Quando comparada com dados observados, os resultados mostram que o modelo é capaz de prever satisfatoriamente as distribuições superficiais e na coluna de água: da temperatura, dos nutrientes, do oxigénio e da clorofila-a. Os resultados evidenciam o crucial papel desempenhado pelos processos físicos no aumento de fitoplâncton que ocorre ao longo de uma estreita área da costa norte portuguesa, mostrando a estreita ligação entre a distribuição costeira de fitoplâncton e a distribuição costeira de temperatura, à superfície. A produtividade do fitoplâncton que ocorre nas águas costeiras não só é atribuída à disponibilidade de nutrientes mas também à intensidade luminosa. A luz e a intensidade dos ventos de norte/noroeste são os factores chave no controlo dos blooms de fitoplâncton observados nesta região de afloramento, sobretudo no Verão, nomeadamente na camada de mistura pouco profunda e junto à nutriclina. Deste modo os modelos numéricos em associação com dados in situ e imagens de satélite poderão ser considerados uma excelente ferramenta para a análise e previsão de cenários, presentes e futuros, de acções praticadas sobre o meio ambiente.