c-KIT expression enhances the leukemogenic potential of 32D cells

The growth of human leukemic cells in culture and in vivo is dependent upon the presence of hematopoietic growth factors. Most populations of human leukemic acute myeloblastic leukemia (AML) cells express c-Kit on their surface and respond to Kit ligand (KL) in culture. To determine if this interaction was of potential significance in vivo we used a mouse model system. 32D cells, a murine IL-3-dependent myeloid cell line, were rendered KL responsive by transfection of the murine c-Kit. After injection of 32D or 32D-Kit cells into syngeneic hosts, animals bearing 32D-Kit cells, but not 32D cells, became moribund and were killed. These animals had circulating leukemic blast cells, infiltration of bone marrow, spleen, brain, liver, lung, and kidney. Cells recovered from some of the animals continued to be dependent upon IL-3 or KL for growth while in other cases the cells were factor independent. This model illustrates that the constitutive expression of c-Kit enhances the leukemic potential of 32D cells. The model will be useful for studying the progression of leukemia in vivo and testing whether interruption of the interaction of Kit and KL can affect the growth of leukemic cells.     

Granularity in ion-beam-sputtered TiO(2) films

TiO(2)-SiO(2) multilayer dielectric stacks that were deposited by ion-beam sputtering were found to have a granular structure. The grains were in the TiO(2) layers rather than in the SiO(2) layers. Their formation was due to the heating of the film during deposition. When the apparatus was modified to reduce the substrate temperature, the granularity was eliminated.     

PAK4, a novel effector for Cdc42Hs, is implicated in the reorganization of the actin cytoskeleton and in the formation of filopodia

The GTPases Rac and Cdc42Hs control diverse cellular functions. In addition to being mediators of intracellular signaling cascades, they have important roles in cell morphogenesis and mitogenesis. We have identified a novel PAK-related kinase, PAK4, as a new effector molecule for Cdc42Hs. PAK4 interacts only with the activated form of Cdc42Hs through its GTPase-binding domain (GBD). Co-expression of PAK4 and the constitutively active Cdc42HsV12 causes the redistribution of PAK4 to the brefeldin A-sensitive compartment of the Golgi membrane and the subsequent induction of filopodia and actin polymerization. Importantly, the reorganization of the actin cytoskeleton is dependent on PAK4 kinase activity and on its interaction with Cdc42Hs. Thus, unlike other members of the PAK family, PAK4 provides a novel link between Cdc42Hs and the actin cytoskeleton. The cellular locations of PAK4 and Cdc42Hs suggest a role for the Golgi in cell morphogenesis.     

Differential activation of ERK and JNK mitogen-activated protein kinases by Raf-1 and MEKK

Growth factors activate mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinases (ERKs) and Jun kinases (JNKs). Although the signaling cascade from growth factor receptors to ERKs is relatively well understood, the pathway leading to JNK activation is more obscure. Activation of JNK by epidermal growth factor (EGF) or nerve growth factor (NGF) was dependent on H-Ras activation, whereas JNK activation by tumor necrosis factor alpha (TNF-alpha) was Ras-independent. Ras activates two protein kinases, Raf-1 and MEK (MAPK, or ERK, kinase) kinase (MEKK). Raf-1 contributes directly to ERK activation but not to JNK activation, whereas MEKK participated in JNK activation but caused ERK activation only after overexpression. These results demonstrate the existence of two distinct Ras-dependent MAPK cascades--one initiated by Raf-1 leading to ERK activation, and the other initiated by MEKK leading to JNK activation.     

Fabrication of optically nonlinear semiconductor mirrors for modelocking of neodymium-doped fiber lasers

A recipe is given for an all-semiconductor, optically nonlinear mirror for passive modelocking of neodymium-doped fiber lasers. InGaAs-GaAs quantum wells (QWs) are used for a saturable absorber. It was empirically found that the best modelocked operation occurred when QWs had a heavy-hole excitonic absorption peak at a wavelength about 10-20 nm longer than the lasing wavelength. An AlAs-GaAs quarter-wavelength stack is used for a rear high reflector. Nonlinear mirrors are etched after epitaxial growth so that the lasing wavelength coincides with a reflectance maximum, giving maximum optical modulation. Nonlinear mirrors are implanted with large doses (2/spl times/10/sup 15//cm/sup 2/) of hydrogen ions in order to increase electron-hole recombination rates, speed material recovery, increase saturation intensity, and inhibit spontaneous Q-switching. Studies of material growth by molecular beam epitaxy and metal-organic chemical vapor deposition (MOCVD) are reported. Modelocked fiber lasers exhibited a pseudorandom output that was produced by several optical pulses that circulated simultaneously in the fiber. It was found that a very high reflectance mirror is not required for lasing or modelocking. To our knowledge, this is the first report of the use of a nonlinear mirror grown by MOCVD to modelock a neodymium fiber laser.     

Differential sensitivity of human myeloma cell lines and normal bone marrow colony forming cells to a recombinant diphtheria toxin-interleukin 6 fusion protein

The cytotoxicity of a recombinant interleukin 6 (IL-6)-diphtheria toxin (DT) fusion protein towards human myeloma cell lines was investigated. DAB389-IL-6 inhibited protein synthesis and methylcellulose colony formation by U266 myeloma cells. In the clonogenic assay, the fusion protein approached the level of cytotoxicity achieved by native DT. The specificity of killing by DAB389-IL-6 was demonstrated by inhibition of cytotoxicity by a molar excess of free rhIL-6. The effect of DAB389-IL-6 on colony formation by six OCI-My cell lines was assessed. Similar to U266 cells, colony growth by the OCI-My 5 and -My 2 cell lines was inhibited in a simple dose dependent manner. However, a biphasic effect was observed for the IL-6 dependent OCI-My 4 cells; DAB389-IL-6 stimulated colony formation at low (< or = 10(-11) M) concentrations, yet was inhibitory at higher doses. Three other cell lines whose growth was not altered by IL-6 were relatively unaffected by DAB389-IL-6, despite their sensitivity to native DT. Flow cytometric analysis for IL-6 receptor expression using phycoerythrin-conjugated IL-6 demonstrated specific binding sites on both DAB389-IL-6 sensitive and certain insensitive cell lines, suggesting that other factors in addition to the expression of IL-6 receptors are involved in killing by the fusion toxin. Despite evidence for a role of IL-6 in myeloid cell development, normal bone marrow was insensitive to the IL-6 fusion toxin. In cultures containing both normal bone marrow and U266 cells DAB389-IL-6 effectively inhibited the growth of U266 myeloma colonies but had little effect on normal bone marrow erythroid, granulocyte and mixed erythroid/granulocyte colony growth. From these experiments we conclude that DAB389-IL-6 is specifically cytotoxic towards a subset of IL-6-responsive human myeloma cell lines and may be useful, in some cases, in the selective elimination of tumour cells from mixed populations of normal and malignant cells.     

A control and management network for wireless ATM systems

This paper describes the design of a control and management network (orderwire) for a mobile wireless Asynchronous Transfer Mode (ATM) network. This mobile wireless ATM network is part of the Rapidly Deployable Radio Network (RDRN). The orderwire system consists of a packet radio network which overlays the mobile wireless ATM network. Each network element in this network uses Global Positioning System (GPS) information to control a beamforming antenna subsystem which provides for spatial reuse. This paper also proposes a novel Virtual Network Configuration (VNC) algorithm for predictive network configuration. A mobile ATM Private Network–Network Interface (PNNI) based on VNC is also discussed. Finally, as a prelude to the system implementation, results of a Maisie simulation of the orderwire system are discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

A range-resolved Doppler imaging sensor based on fiber lasers

We have developed a unique eyesafe (1.5 μm) laser with a pseudorandom pulse output. The laser is based on a mode-locked pulse format; the pulse pattern repeats over a time consistent with the cavity round-trip period but appears random within that period. The pseudorandom pulse format occurs when we overdrive a passive mode-locking mirror in a long fiber laser. This laser is used for generating range-resolved Doppler images