Acoustic and vibration isolation for a gravity gradiometer.

Vibration in the audio frequency band affects the performance of rotating gravity gradiometers used for airborne mineral exploration. This is probably due to translation to rotation coupling inside the gradiometer platform. It was found that the DC gravity gradient signal was proportional to the square of the third time derivative of position, or jerk squared. The demanding airborne environment for such instrumentation demands a light weight broadband acoustic shield and vibration isolator. This paper presents the design principles for such an isolator, based on vibration isolated spherical shell structures. Performance data are presented as well as flight test data that demonstrated a 14% gravity gradient noise reduction compared with an unshielded instrument.

Rotational isolation with neutrally buoyant suspension.

Rotational vibration isolation is of critical importance for many airborne instrumentation applications. Such isolators require very low frequency isolation for the rotational degrees of freedom combined with translational rigidity and negligible translation to rotation coupling. This paper describes a vibration isolator using neutrally buoyant flotation to provide high translation rigidity combined with very low rotational rigidity. The isolator reduces the rotational vibration at all frequencies above its resonance (0.18 ± 0.01 Hz) and has a large dynamic range (±30°) suitable for airborne surveying. Viscous, inviscid, and mechanical coupling inside the isolator have been analyzed. A recent fixed wing flight test shows the isolator reducing the rotational vibration by more than a factor of 1000 at frequencies above 10 Hz.

Angular instability in high optical power suspended cavities.

Advanced gravitational wave detectors use suspended test masses to form optical resonant cavities for enhancing the detector sensitivity. These cavities store hundreds of kilowatts of coherent light and even higher optical power for future detectors. With such high optical power, the radiation pressure effect inside the cavity creates a sufficiently strong coupling between test masses whose dynamics are significantly altered. The dynamics of two independent nearly free masses become a coupled mechanical resonator system. The transfer function of the local control system used for controlling the test masses is modified by the radiation pressure effect. The changes in the transfer function of the local control systems can result in a new type of angular instability which occurs at only 1.3% of the Sidles-Sigg instability threshold power. We report the experimental results on a 74 m suspended cavity with a few kilowatts of circulating power, for which the power to mass ratio is comparable to the current Advanced LIGO. The radiation pressure effect on the test masses behaves like an additional optical feedback with respect to the local angular control, potentially making the mirror control system unstable. When the local angular control system is optimised for maximum stability margin, the instability threshold power increases from 4 kW to 29 kW. The system behaviour is consistent with our simulation, and the power dependent evolution of both the cavity soft and hard mode is observed. We show that this phenomenon is likely to significantly affect the proposed gravitational wave detectors that require very high optical power.

Narrowing the filter-cavity bandwidth in gravitational-wave detectors via optomechanical interaction.

We propose using optomechanical interaction to narrow the bandwidth of filter cavities for achieving frequency-dependent squeezing in advanced gravitational-wave detectors, inspired by the idea of optomechanically induced transparency. This can allow us to achieve a cavity bandwidth on the order of 100 Hz using small-scale cavities. Additionally, in contrast to a passive Fabry-Pérot cavity, the resulting cavity bandwidth can be dynamically tuned, which is useful for adaptively optimizing the detector sensitivity when switching amongst different operational modes. The experimental challenge for its implementation is a stringent requirement for very low thermal noise of the mechanical oscillator, which would need a superb mechanical quality factor and a very low temperature. We consider one possible setup to relieve this requirement by using optical dilution to enhance the mechanical quality factor.

Near-self-imaging cavity for three-mode optoacoustic parametric amplifiers using silicon microresonators.

Three-mode optoacoustic parametric amplifiers (OAPAs), in which a pair of photon modes are strongly coupled to an acoustic mode, provide a general platform for investigating self-cooling, parametric instability and very sensitive transducers. Their realization requires an optical cavity with tunable transverse modes and a high quality-factor mirror resonator. This paper presents the design of a table-top OAPA based on a near-self-imaging cavity design, using a silicon torsional microresonator. The design achieves a tuning coefficient for the optical mode spacing of 2.46 MHz/mm. This allows tuning of the mode spacing between amplification and self-cooling regimes of the OAPA device. Based on demonstrated resonator parameters (frequencies ∼400 kHz and quality-factors ∼7.5×10(5) we predict that the OAPA can achieve parametric instability with 1.6 µW of input power and mode cooling by a factor of 1.9×10(4) with 30 mW of input power.

Novel Euler-LaCoste linkage as a very low frequency vertical vibration isolator.

LaCoste linkage vibration isolators have shown excellent performance for ultra-low frequency vertical vibration isolation. However, such isolators depend on the use of conventional pre-stressed coil springs, which suffer from creep. Here, we show that compressional Euler springs can be configured to create a stable tension unit for use in a LaCoste structure. In a proof of concept experiment, we demonstrate a vertical resonance frequency of 0.15 Hz in an Euler-LaCoste configuration with 200 mm height. The system enables the use of very low creep maraging steel as spring elements to eliminate the creep while minimising spring mass and reducing the effect of parasitic resonances. Larger scale systems with optimized Euler spring boundary conditions should achieve performance suitable for applications on third generation gravitational wave detectors such as the proposed Einstein telescope.

Compact vibration isolation and suspension for Australian International Gravitational Observatory: performance in a 72 m Fabry Perot cavity.

This paper describes the first demonstration of vibration isolation and suspension systems, which have been developed with view to application in the proposed Australian International Gravitational Observatory. In order to achieve optimal performance at low frequencies new components and techniques have been combined to create a compact advanced vibration isolator structure. The design includes two stages of horizontal preisolation and one stage of vertical preisolation with resonant frequencies approximately 100 mHz. The nested structure facilitates a compact design and enables horizontal preisolation stages to be configured to create a superspring configuration, where active feedback can enable performance close to the limit set by seismic tilt coupling. The preisolation stages are combined with multistage three-dimensional (3D) pendulums. Two isolators suspending mirror test masses have been developed to form a 72 m optical cavity with finesse approximately 700 in order to test their performance. The suitability of the isolators for use in suspended optical cavities is demonstrated through their ease of locking, long term stability, and low residual motion. An accompanying paper presents the local control system and shows how simple upgrades can substantially improve residual motion performance.

Feedback control of thermal lensing in a high optical power cavity.

This paper reports automatic compensation of strong thermal lensing in a suspended 80 m optical cavity with sapphire test mass mirrors. Variation of the transmitted beam spot size is used to obtain an error signal to control the heating power applied to the cylindrical surface of an intracavity compensation plate. The negative thermal lens created in the compensation plate compensates the positive thermal lens in the sapphire test mass, which was caused by the absorption of the high intracavity optical power. The results show that feedback control is feasible to compensate the strong thermal lensing expected to occur in advanced laser interferometric gravitational wave detectors. Compensation allows the cavity resonance to be maintained at the fundamental mode, but the long thermal time constant for thermal lensing control in fused silica could cause difficulties with the control of parametric instabilities.

Compensation of strong thermal lensing in high-optical-power cavities.

In an experiment to simulate the conditions in high optical power advanced gravitational wave detectors, we show for the first time that the time evolution of strong thermal lenses follows the predicted infinite sum of exponentials (approximated by a double exponential), and that such lenses can be compensated using an intracavity compensation plate heated on its cylindrical surface. We show that high finesse approximately 1400 can be achieved in cavities with internal compensation plates, and that mode matching can be maintained. The experiment achieves a wave front distortion similar to that expected for the input test mass substrate in the Advanced Laser Interferometer Gravitational Wave Observatory, and shows that thermal compensation schemes are viable. It is also shown that the measurements allow a direct measurement of substrate optical absorption in the test mass and the compensation plate.

Parametric instabilities and their control in advanced interferometer gravitational-wave detectors.

A detailed simulation of Advanced LIGO test mass optical cavities shows that parametric instabilities will excite 7 acoustic modes in each fused silica test mass, with parametric gain R up to 7 and only 1 acoustic mode with R approximately 2 for alternative sapphire test masses. Fine-tuning of the test mass radii of curvature causes the instabilities to sweep through various modes with R as high as approximately 2000. Sapphire test mass cavities can be tuned to completely eliminate instabilities using thermal g-factor tuning. In the case of fused silica test mass, instabilities can be minimized but not eliminated.

The bone morphogenic protein antagonist gremlin regulates proximal-distal patterning of the lung.

The proximal-distal patterning of lung epithelium involves a complex series of signaling and transcriptional events resulting in the programmed differentiation of highly specialized cells for gas exchange and surfactant protein expression essential for postnatal lung function. The BMP signaling pathway has been shown to regulate cellular differentiation in the lung as well as other tissues. In this report, we show that the can family of related BMP antagonists, including gremlin, cer-1, PRDC, and Dan are expressed in the lung during embryonic development with gremlin expression observed in the proximal airway epithelium. The role of gremlin in lung development was explored by overexpressing it in the distal lung epithelium of transgenic mice using the human SP-C promoter. SP-C/gremlin transgenic mice exhibited a disruption of the proximal-distal patterning found in the airways of the mammalian lung. Expanded expression of the proximal epithelial cell markers CC10 and HFH-4 (Foxj1) was observed in the distal regions of transgenic lungs. Furthermore, smooth muscle alpha-actin expression was observed surrounding the distal airways of SP-C/gremlin mice, indicating a proximalization of distal lung tubules. These data suggest that gremlin plays an important role in lung morphogenesis by regulating the proximal-distal patterning of the lung during development.

The transforming activity of tumour cell DNA.

Transfection of DNA isolated from a subset of human tumour cells was shown in the early 1980s to be capable of inducing transformation in NIH3T3 cells. This review summarizes the approaches and results of the efforts to measure the transforming activity of genomic DNA and discusses the results in the light of current concerns about the potential hazard associated with DNA contamination of biological material produced by cultured transformed cells.

Expression of functional formyl peptide receptors by human astrocytoma cell lines.

Activation of astrocytes is important in the pathogenesis of a variety of diseases in the central nervous system, such as infection and neurodegeneration. We found that the bacterial chemotactic peptide, N-formyl-methionyl-leucyl-phenylalanine (fMLF) induced potent migration and Ca(2+) mobilization in human astrocytoma cell lines. The effect of fMLF was pertussis toxin-sensitive, suggesting the involvement of seven transmembrane, G protein-coupled receptor(s) for fMLF. Scatchard analyses revealed that astrocytoma cell lines express both high- and low-affinity binding sites for [3H]fMLF. RT-PCR confirmed the expression of transcripts of fMLF receptors, the high-affinity FPR and the low-affinity FPRL1 by these cells. Both fMLF and F peptide, a synthetic peptide domain of HIV-1 envelope protein which specifically activates FPRL1, increased secretion of IL-6 by astrocytoma cells. Our study demonstrates for the first time that FPR and FPRL1 expressed by astrocytoma cell lines are functional, and suggests a molecular basis for the involvement of these receptors in host defense in the brain.

Suppression of the Ewing's sarcoma phenotype by FLI1/ERF repressor hybrids.

Fusion of the 5' half of the Ewing's sarcoma (ES) gene EWS with the DNA-binding domain of several transcription factors has been detected in many human tumors. The t(11;22)(q24;q12) chromosomal translocation is specifically linked to ES and primitive neuroectodermal tumors and results, in the majority of cases, in the fusion of the amino terminus of the EWS gene to the carboxyl-terminal DNA-binding domain of the FLI1 gene. The chimeric protein has been shown to be oncogenic, a potent transcriptional activator, and necessary for the maintenance of the Ewing's phenotype, making it an attractive target for gene therapy. In this study, we demonstrate that the ES transformed phenotype can be suppressed by chimeric transcriptional repressors containing the DNA-binding domain of FLI1 and the regulatory and repressor domain of ERF, a transcription suppressor and member of the ets gene family. The hybrid repressor is expressed at levels comparable with EWS/FLI1, does not affect EWS/FLI1 expression, and exhibits similar DNA-binding specificity but suppresses transcriptional activity. The FLI1/ERF repressor, like the wild-type ERF, is regulated by mitogen-activated protein kinase-dependent subcellular localization. Our data suggest that transformation by EWS/FLI1 may partially be due to activation of specific EWS/FLI1-regulated genes involved in cell proliferation.

Cloning of the murine Drm gene (Cktsf1b1) and characterization of its oncogene suppressible promoter.

The Drm gene, first identified in rat cells in our laboratory, appears to play a significant role in early embryo patterning and limb bud development. We have now isolated mouse Drm (mDrm) cDNA as well as genomic DNA clones and have mapped the Drm gene (Cktsf1b1) to murine chromosome 2. Cktsf1b1 is regulated in a tissue specific fashion and is expressed only in nontransformed mouse cells or primary fibroblasts in culture, but not in established transformed or tumor-derived mouse cell lines. The major transcription start sites map to within 69 bp upstream of the initiating ATG. A promoter was contained in the -214 to +1 bp 5' flanking region, and promoter/reporter constructs showed 10-fold higher activity than control in REF-1 (rat), A31 (mouse) and CHO (hamster) cells. The region contains a TATA sequence and multiple potential transcription factor binding sites. Promoter activity was dose-dependently inhibited by cotransfection with either ras or mos oncogenes, but oncogene inhibition was reversed and the overall activity increased when cells were treated with the MAP kinase kinase (MKK) inhibitor PD98059. An NF-1 and Yi-like site, identified in the minimal promoter region, showed different mobility shift patterns when normal and transformed cell nuclear extracts are compared. Mutation of the NF-1 site reduced Cktsf1b1 promoter activity 25%, while mutation of the Yi-like site destroyed all the activity. Our results indicate that the expression of Cktsf1b1, a gene associated with early development and cell transformation, is sensitive to MKK levels and may be regulated via multiple transcription factor complexes.

DRM/GREMLIN (CKTSF1B1) maps to human chromosome 15 and is highly expressed in adult and fetal brain.

We have mapped and characterized the human homolog of Drm/Gremlin (CKTFS1B1), a member of a family of BMP antagonists that have been linked to both developmental and transformation-related functions. By screening a human cDNA library, we isolated a 3.3-kb cDNA containing the 552-bp region encoding the human DRM protein. CKTFS1B1 was localized on human chromosome 15q13--> q15 by somatic cell hybrid analysis and, more precisely, using radiation hybrids, to a region of markers linked to SGNE1, secretory granule neuroendocrine protein 1 and RYR3, the ryanodyne receptor 3. Northern blot analysis showed the presence of a single DRM-specific mRNA expressed in different human tissues, including brain, ovary, intestine and colon. In the brain, DRM expression is associated with the region localized around the internal capsule in the large subcortical nuclei. DRM appears to be predominantly expressed in normal cells and tissues, including normal neurons, astrocytes and fibroblasts. Interestingly, we detected DRM expression in normal cells obtained from several patients, but not in tumor cell lines established from the same patients. The data suggest that down-regulation of DRM is associated with tumor progression, and support the hypothesis that human DRM may play an important role during both neuroembryological development and carcinogenesis.

Biosynthesis, post-translation modification, and functional characterization of Drm/Gremlin.

Down-regulated by mos (Drm)/Gremlin is a highly conserved protein whose properties and expression pattern suggest a role in early development, tissue-specific differentiation, and cell transformation. We have investigated the biosynthesis and processing of Drm expressed endogenously in rat fibroblasts or overexpressed following transient or stable transfection. Analysis of metabolically labeled cells revealed that Drm exists in secreted and cell-associated forms that exhibit similar mobilities in SDS-polyacrylamide gel electrophoresis. Protein analysis indicated that Drm is present in two major species: a slow migrating glycosylated form and a nonglycosylated form. Both forms of Drm are able to undergo phosphorylation. Drm is released into the media within 30 min of synthesis and is detectable for up to 4-5 h, whereas the cell-associated form has a half-life of about 1 h. Confocal immunofluorescent microscopy indicates that Drm is present both on the external surface of expressing cells, as well as within the endoplasmic reticulum and the Golgi. Both glycosylated and nonglycosylated forms of Drm exhibit identical distributions and are able to antagonize bone morphogenetic protein signaling. Like the soluble form, the cell-associated forms are capable of binding (125)I-bone morphogenetic protein-4. These properties are consistent with a role for Drm in interfering with signaling and indicate that Drm may act at the cell surface during tissue development and transformation.

FLI-1 is a suppressor of erythroid differentiation in human hematopoietic cells.

The FLI-1 oncogene, a member of the ETS family of transcription factors, is associated with both normal and abnormal hematopoietic cell growth and lineage-specific differentiation. We have previously shown that overexpression of FLI-1 in pluripotent human hematopoietic cells leads to the induction of a megakaryocytic phenotype. In this report we show that FLI-1 also acts as an inhibitor of erythroid differentiation. Following the induction of erythroid differentiation, pluripotent cells express reduced levels of FLI-1. In contrast, when FLI-1 is overexpressed in these cells, the levels of erythroid markers are reduced. The ability of FLI-1 overexpressing cells to respond to erythroid-specific inducers such as hemin and Ara-C is also inhibited, and the uninduced cells show a reduced level of the erythroid-associated GATA-1 transcription factor mRNA. Furthermore, expression of a GATA-1 promoter-driven reporter construct in K562 cells is inhibited by co-transfection with a construct expressing FLI-1. Our results support the hypothesis that FLI-1 can act both positively and negatively in the regulation of hematopoietic cell differentiation, and that inhibition of GATA-1 expression may contribute to FLI-1-mediated inhibition of erythroid differentiation.

Ets and retroviruses - transduction and activation of members of the Ets oncogene family in viral oncogenesis.

Studies of retroviral-induced oncogenesis in animal systems led to the initial discovery of viral oncogenes and their cellular homologs, and provided critical insights into their role in the neoplastic process. V-ets, the founding member of the ETS oncogene family, was originally identified as part of the fusion oncogene encoded by the avian acute leukemia virus E26 and subsequent analysis of virus induced leukemias led to the initial isolation of two other members of the ETS gene family. PU.1 was identified as a target of insertional activation in the majority of tumors induced by the murine Spleen Focus Forming virus (SFFV), while fli-1 proved to be the target of Friend murine leukemia virus (F-MuLV) in F-MuLV induced erythroleukemia, as well as that of the 10A1 and Graffi viruses. The common features of the erythroid and myeloid diseases induced by these viruses provided the initial demonstration that these and other members of the ETS family play important roles in hematopoietic development as well as disease. This review provides an overview of the role of ETS genes in retrovirally induced neoplasia, their possible mechanisms of action, and how these viral studies relate to current knowledge of the functions of these genes in hematopoiesis.

A mouse fibroblast line cycles between monolayer and spheroid forms, regulates Met and HGF expression, and releases an attachment and growth-promoting substance.

A subline of mesoderm-derived mouse NIH3T3 fibroblasts was selected for its ability to proliferate in serum-free media. This cell line (SFDH) grows as a monolayer at low density and spontaneously forms dense, multicellular spheroids at high density. Spheroid formation can also be induced by the addition of dexamethasone, polybrene, or heparin. Spheroids eventually detach from the substrate, but will reattach and re-form monolayers when transferred to fresh culture vessels and media, repeating the cycle again upon reaching high density. Thin section analysis of spheroids shows morphologically-distinct regions of cells, including an attenuated outer surface and a cuboidal interior with occasional lumen-like areas. Over time in culture, spheroids express increasing levels of met, the Met ligand-SF/HGF and cytokeratin, an epithelial marker, in comparison to monolayers. Both monolayer and spheroid-derived cells are rapidly tumorigenic in nude mice. Media conditioned by SFDH cells contain factors that stimulate growth and attachment of a variety of tumorigenic and non-tumorigenic cell lines, inducing cells to divide in serum-free media for up to 14 days when plated on tissue culture-treated and nontreated plastic surfaces pre-coated with SFDH conditional media. The growth-stimulating activity fractionates as a single peak over a sepharose column in the presence of 6 m urea, and sediments as a high molecular weight complex. Growth-stimulating activity can be neutralized by several antisera specific for hepatocyte growth factor, and the same sera recognize a novel approximately 37 kD protein in active supernatants. The cyclic, continuous nature of alternating monolayer and spheroid forms makes this cell line appropriate for studying changing gene expression patterns in progressive cell-cell/cell-matrix interactions.