Diphtheria toxin fused to granulocyte-macrophage colony-stimulating factor eliminates acute myeloid leukemia cells with the potential to initiate leukemia in immunodeficient mice, but spares normal hemopoietic stem cells

We studied the cell kill induced by granulocyte-macrophage colony-stimulating factor (GM-CSF ) fused to Diphtheria Toxin (DT-GM-CSF ) in acute myeloid leukemia (AML) samples and in populations of normal primitive hemopoietic progenitor cells. AML samples from three patients were incubated in vitro with 100 ng/mL DT-GM-CSF for 48 hours, and AML cell kill was determined in a proliferation assay, a clonogenic assay colony-forming unit-AML (CFU-AML) and a quantitative long-term bone marrow (BM) culture ie, the leukemic-cobblestone area forming cell assay (L-CAFC). To measure an effect on cells with in vivo leukemia initiating potential DT-GM-CSF exposed AML cells were transplanted into immunodeficient mice. In two out of three samples it was shown that all AML subsets, including those with long-term abilities in vivo (severe combined immunodeficient mice) and in vitro (L-CAFC assay) were reduced in number by DT-GM-CSF. Cell kill induced by DT-GM-CSF could be prevented by coincubation with an excess of GM-CSF, demonstrating that sensitivity to DT-GM-CSF is specifically mediated by the GM-CSF receptor. Therefore, binding and internalization of GM-CSF probably occur in immature AML precursors of these two cases of AML. The third AML sample was not responsive to either GM-CSF or DT-GM-CSF. The number of committed progenitors of normal bone marrow (burst-forming unit-erythroid, colony-forming unit granulocyte- macrophage, and cobble stone area forming cell [CAFC] week 2) and also the number of cells with long-term repopulating ability, assayed as week 6 CAFC, were unchanged after exposure to DT-GM-CSF (100 ng/mL, 48 hours). These studies show that DT-GM-CSF may be used to eliminate myeloid leukemic cells with long-term potential in vitro and in immunodeficient mice, whereas normal hemopoietic stem cells are spared.     

A 1.8-mW CMOS ΣΔ modulator with integrated mixer forA/D conversion of IF signals

In this paper, the design of a continuous-time baseband sigma-delta (ΣΔ) modulator with an integrated mixer for intermediate-frequency (IF) analog-to-digital conversion is presented. This highly linear IF ΣΔ modulator digitizes a GSM channel at intermediate frequencies up to 50 MHz. The sampling rate is not related to the input IF and is 13.0 MHz in this design. Power consumption is 1.8 mW from a 2.5-V supply. Measured dynamic range is 82 dB, and third-order intermodulation distortion is -84 dB for two -6-dBV IF input tones. Two modulators in quadrature configuration provide 200-kHz GSM bandwidth. Active area of a single IF ΣΔ modulator is 0.2 mm² in 0.35-μm CMOS     

An IF-to-Baseband Σ∆ Modulator for AM/FM/IBOC Radio Receivers With a 118 dB Dynamic Range

A single-bit fifth-order complex continuous-time IF-to-baseband SigmaDelta modulator for AM/FM/IBOC receivers is presented. The input IF is 10.7 MHz and the sampling frequency is 41.7 MHz. The modulator achieves a dynamic range of 118dB in AM mode (3 kHz BW), 98dB in FM mode (200 kHz BW), and 86dB in IBOC mode (500 kHz BW). The modulator's high dynamic range enables the realization of an AM radio receiver without a VGA and without an AM channel-selection filter, thereby reducing system complexity and cost. The elimination of the VGA also improves the sensitivity and the overall noise figure of the receiver. The modulator's spurious free dynamic range is 88 dB in the bandwidth from 25 to 525 kHz. The IM2 distance is 92 dB, and the IM3 distance is 91 dB. The ADC was fabricated in a one-poly five-metal 0.18-mum CMOS process with an active area of 6.0mm². It consumes 210 mW from a 1.8-V supply     

Frequency analysis of human primitive haematopoietic stem cell subsets using a cobblestone area forming cell assay

Stroma-dependent long-term bone marrow cultures (LTBMC) assay the ability of primitive haematopoietic stem cells (HSC) for long-term production of clonable progenitors. We have developed a limiting dilution type LTBMC assay allowing frequency analysis of transiently repopulating HSC and long-term culture initiating cells (LTC-IC) without the necessity to replate large numbers of wells. Normal or 5-FU-treated Ficoll bone marrow cells (BMC), or BMCs sorted on CD34 or HLA-DR expression, or Rh123 retention, (input range 40-70,000 CFU-GM/BFU-E/10(5) cells) were plated at limiting dilution on unirradiated adherent layers formed by a novel murine preadipose cell line (FBMD-1). The percentage of wells with at least one phase-dark haematopoietic clone (cobblestone area, CA) beneath the stromal layer was weekly determined for at least 8 weeks, and CA-forming cell (CAFC) frequencies were calculated using Poisson statistics. Parallel LTBMCs of the same samples were weekly assessed for supernate CFU-GM/BFU-E production. Weekly addition of rhIL-3 with rhG-CSF supported a high average clonogenic output per CA and dramatically increased CA size, but did not significantly alter the apparent CAFC frequency. The generation of CFU-GM per CA was constant over a period of 6 weeks with weekly means of eight normal BM samples, ranging between 5-16. At week 6 the mean CAFC frequency was 29 (1 SEM, 8.8)/10(5). Early appearing CAFC were highly sensitive to 5-FU, and were contained over the full Rh123 and HLA-DR fluorescence profile of CD34pos cells, whereas CAFC week 5-8 were predominantly contained in the CD34pos Rh123dull HLA-DRlow fraction in agreement with previously reported LTC-IC characteristics. In conclusion, the CAFC assay enumerates LTC-IC using a direct visual endpoint and allows study of LTC-IC heterogeneity with respect to progenitor cell generation per stem cell clone in various haematologic diseases.     

A cascaded continuous-time /spl Sigma//spl Delta/ Modulator with 67-dB dynamic range in 10-MHz bandwidth

This paper presents the design of a 2-2 cascaded continuous-time sigma-delta modulator. The cascaded modulator comprises two stages with second-order continuous-time resonator loopfilters, 4-bit quantizers, and feedback digital-to-analog converters. The digital noise cancellation filter design is determined using continuous-time to discrete-time transformation of the sigma-delta loopfilter transfer functions. The required matching between the analog and digital filter coefficients is achieved by means of simple digital calibration of the noise cancellation filter. Measurement results of a 0.18-/spl mu/m CMOS prototype chip demonstrate 67-dB dynamic range in a 10-MHz bandwidth at 8 times oversampling for a single continuous-time cascaded modulator. Two cascaded modulators in quadrature configuration provide 20-MHz aggregate bandwidth. Measured anti-alias suppression is over 50 dB for input signals in the band from 150 to 170 MHz around the sampling frequency of 160 MHz.     

A quadrature data-dependent DEM algorithm to improve imagerejection of a complex ΣΔ modulator

A dynamic element matching (DEM) algorithm is presented that is controlled by the quadrature output data of a complex sigma-delta modulator. This DEM technique is used to correct the gain and phase errors between the circuits in the in-phase and quadrature-phase feedback paths of the modulator. The key feature of this DEM technique is that it does not cause leakage of high-frequency quantization noise in the signal band, as encounters with the periodic or pseudorandom DEM techniques. No test signal is required to measure the gain and phase errors, and as the DEM circuit is operating continuously, it compensates for changes in, e.g., temperature and supply voltage. A 0.35-μm CMOS prototype chip has been designed to test the DEM circuit. A batch of 38 measured samples shows a typical mismatch-independent image rejection ratio of 63 dB with DEM     

A 56 mW Continuous-Time Quadrature Cascaded Σ∆ Modulator With 77 dB DR in a Near Zero-IF 20 MHz Band

A quadrature cascaded modulator with continuous-time loop filters is presented for a digital multi-stream FM radio receiver. The ADC achieves a dynamic range of 77 dB and 20 MHz bandwidth centered on an intermediate frequency of 10.5 MHz and is sampling at 340 MHz. The cascaded modulator comprises programmable analog second-order quadrature filters and a digital quadrature noise cancellation filter. The 0.5 chip in 90 nm CMOS consumes 56 mW from a 1.2 V supply.