Significantly improved PCR-based clonality testing in B-cell malignancies by use of multiple immunoglobulin gene targets: Report of the BIOMED-2 Concerted Action BHM4-CT98-3936.

Polymerase chain reaction (PCR) assessment of clonal immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements is an important diagnostic tool in mature B-cell neoplasms. However, lack of standardized PCR protocols resulting in a high level of false negativity has hampered comparability of data in previous clonality studies. In order to address these problems, 22 European laboratories investigated the Ig/TCR rearrangement patterns as well as t(14;18) and t(11;14) translocations of 369 B-cell malignancies belonging to five WHO-defined entities using the standardized BIOMED-2 multiplex PCR tubes accompanied by international pathology panel review. B-cell clonality was detected by combined use of the IGH and IGK multiplex PCR assays in all 260 definitive cases of B-cell chronic lymphocytic leukemia (n¼56), mantle cell lymphoma (n¼54), marginal zone lymphoma (n¼41) and follicular lymphoma (n¼109). Two of 109 cases of diffuse large B-cell lymphoma showed no detectable clonal marker. The use of these techniques to assign cell lineage should be treated with caution as additional clonal TCR gene rearrangements were frequently detected in all disease categories. Our study indicates that the BIOMED-2 multiplex PCR assays provide a powerful strategy for clonality assessment in B-cell malignancies resulting in high Ig clonality detection rates particularly when IGH and IGK strategies are combined.

Th2 lymphoproliferative disorder of LatY136F mutant mice unfolds independently of TCR-MHC engagement and is insensitive to the action of Foxp3+ regulatory T cells.

Mutant mice where tyrosine 136 of linker for activation of T cells (LAT) was replaced with a phenylalanine (Lat(Y136F) mice) develop a fast-onset lymphoproliferative disorder involving polyclonal CD4 T cells that produce massive amounts of Th2 cytokines and trigger severe inflammation and autoantibodies. We analyzed whether the Lat(Y136F) pathology constitutes a bona fide autoimmune disorder dependent on TCR specificity. Using adoptive transfer experiments, we demonstrated that the expansion and uncontrolled Th2-effector function of Lat(Y136F) CD4 cells are not triggered by an MHC class II-driven, autoreactive process. Using Foxp3EGFP reporter mice, we further showed that nonfunctional Foxp3(+) regulatory T cells are present in Lat(Y136F) mice and that pathogenic Lat(Y136F) CD4 T cells were capable of escaping the control of infused wild-type Foxp3(+) regulatory T cells. These results argue against a scenario where the Lat(Y136F) pathology is primarily due to a lack of functional Foxp3(+) regulatory T cells and suggest that a defect intrinsic to Lat(Y136F) CD4 T cells leads to a state of TCR-independent hyperactivity. This abnormal status confers Lat(Y136F) CD4 T cells with the ability to trigger the production of Abs and of autoantibodies in a TCR-independent, quasi-mitogenic fashion. Therefore, despite the presence of autoantibodies causative of severe systemic disease, the pathological conditions observed in Lat(Y136F) mice unfold in an Ag-independent manner and thus do not qualify as a genuine autoimmune disorder.

A recursive rule base adjustment algorithm for a fuzzy logic controller

This paper introduces a recursive rule base adjustment to enhance the performance of fuzzy logic controllers. Here the fuzzy controller is constructed on the basis of a decision table (DT), relying on membership functions and fuzzy rules that incorporate heuristic knowledge and operator experience. If the controller performance is not satisfactory, it has previously been suggested that the rule base be altered by combined tuning of membership functions and controller scaling factors. The alternative approach proposed here entails alteration of the fuzzy rule base. The recursive rule base adjustment algorithm proposed in this paper has the benefit that it is computationally more efficient for the generation of a DT, and advantage for online realization. Simulation results are presented to support this thesis. (c) 2005 Elsevier B.V. All rights reserved.

Performance projections and design optimization of planar double gate SOI MOSFETs for logic technology applications

In this paper, by investigating the influence of source/drain extension region engineering (also known as gate-source/drain underlap) in nanoscale planar double gate (DG) SOI MOSFETs, we offer new insights into the design of future nanoscale gate-underlap DG devices to achieve ITRS projections for high performance (HP), low standby power (LSTP) and low operating power (LOP) logic technologies. The impact of high-kappa gate dielectric, silicon film thickness, together with parameters associated with the lateral source/drain doping profile, is investigated in detail. The results show that spacer width along with lateral straggle can not only effectively control short-channel effects, thus presenting low off-current in a gate underlap device, but can also be optimized to achieve lower intrinsic delay and higher on-off current ratio (I-on/I-off). Based on the investigation of on-current (I-on), off-current (I-off), I-on/I-off, intrinsic delay (tau), energy delay product and static power dissipation, we present design guidelines to select key device parameters to achieve ITRS projections. Using nominal gate lengths for different technologies, as recommended from ITRS specification, optimally designed gate-underlap DG MOSFETs with a spacer-to-straggle (s/sigma) ratio of 2.3 for HP/LOP and 3.2 for LSTP logic technologies will meet ITRS projection. However, a relatively narrow range of lateral straggle lying between 7 to 8 nm is recommended. A sensitivity analysis of intrinsic delay, on-current and off-current to important parameters allows a comparative analysis of the various design options and shows that gate workfunction appears to be the most crucial parameter in the design of DG devices for all three technologies. The impact of back gate misalignment on I-on, I-off and tau is also investigated for optimized underlap devices.

Multiple Input Governor Control for a Diesel Generating Set

The paper presents a multiple input single output fuzzy logic governor algorithm that can be used to improve the transient response of a diesel generating set, when supplying an islanded load. The proposed governor uses the traditional speed input in addition to voltage and power factor to modify the fuelling requirements during various load disturbances. The use of fuzzy logic control allows the use of PID type structures that can provide variable gain strategies to account for non-linearities in the system. Fuzzy logic also provides a means of processing other input information by linguistic reasoning and a logical control output to aid the governor action during transient disturbance. The test results were obtained using a 50 kVA naturally aspirated diesel generator testing facility. Both real and reactive load tests were conducted. The complex load test results demonstrate that, by using additional inputs to the governor algorithm, enhanced generator transient speed recovery response can be obtained.

Kir2.2 inward rectifier potassium channels are inhibited by an endogenous factor in Xenopus oocytes independently from the action of a mitochondrial uncoupler.

We previously showed inhibition of Kir2 inward rectifier K+ channels expressed in Xenopus oocytes by the mitochondrial agents carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) and sodium azide. Mutagenesis studies suggested that FCCP may act via phosphatidylinositol 4,5-bisphosphate (PIP2) depletion. This mechanism could be reversible in intact cells but not in excised membrane patches which preclude PIP2 regeneration. This prediction was tested by investigating the reversibility of the inhibition of Kir2.2 by FCCP in intact cells and excised patches. We also investigated the effect of FCCP on Kir2.2 expressed in human embryonic kidney (HEK) cells. Kir2.2 current, expressed in Xenopus oocytes, increased in inside-out patches from FCCP-treated and untreated oocytes. The fraction of total current that increased was 0.79?±?0.05 in control and 0.89?±?0.03 in 10 µM FCCP-treated (P?>?.05). Following “run-up,” Kir2.2 current was re-inhibited by “cramming” inside-out patches into oocytes. Therefore, run-up reflected not reversal of inhibition by FCCP, but washout of an endogenous inhibitor. Kir2.2 current recovered in intact oocytes within 26.5 h of FCCP removal. Injection of oocytes with 0.1 U apyrase completely depleted ATP (P?<?.001) but did not inhibit Kir2.2 and inhibited Kir2.1 by 35% (P?<?.05). FCCP only partially reduced [ATP] (P?<?.001), despite inhibiting Kir2.2 by 75% (P?<?.01) but not Kir2.1. FCCP inhibited Kir2.2 expressed in HEK cells. The recovery of Kir2.2 from inhibition by FCCP requires intracellular components, but direct depletion of ATP does not reproduce the differential inhibitory effect of FCCP. Inhibition of Kir2.2 by FCCP is not unique to Xenopus oocytes. J. Cell. Physiol. 219: 8–13, 2009. © 2008 Wiley-Liss, Inc.