Robert H. Giles

Diagnostic analysis of the Rubinstein-Taybi syndrome: five cosmids should be used for microdeletion detection and low number of protein truncating mutations

Rubinstein-Taybi syndrome (RTS) is a malformation syndrome characterised by facial abnormalities, broad thumbs, broad big toes, and mental retardation. In a subset of RTS patients, microdeletions, translocations, and inversions involving chromosome band 16p13.3 can be detected. We have previously shown that disruption of the human CREB binding protein (CREBBP orCBP) gene, either by these gross chromosomal rearrangements or by point mutations, leads to RTS. CBP is a large nuclear protein involved in transcription regulation, chromatin remodelling, and the integration of several different signal transduction pathways. Here we report diagnostic analysis of CBP in 194 RTS patients, divided into several subsets. In one case the mother is also suspect of having RTS. Analyses of the entireCBP gene by the protein truncation test showed 4/37 truncating mutations. Two point mutations, one 11 bp deletion, and one mutation affecting the splicing of the second exon were detected by subsequent sequencing. Screening theCBP gene for larger deletions, by using different cosmid probes in FISH, showed 14/171 microdeletions. Using five cosmid probes that contain the entire gene, we found 8/89 microdeletions of which 4/8 were 5′ or interstitial. This last subset of microdeletions would not have been detected using the commonly used 3′ probe RT1, showing the necessity of using all five probes.

Detection of CBP rearrangements in acute myelogenous leukemia with t(8;16)

The CREB-binding protein (CBP) is a large nuclear protein that regulates many signal transduction pathways and is involved in chromatin-mediated transcription. The translocation t(8;16)(p11;p13.3) consistently disrupts two genes: the CBP gene on chromosome band 16p13.3 and the MOZ gene on chromosome band 8p11. Although a fusion of these two genes as a result of the translocation is expected, attempts at detecting the fusion transcript by reverse transcriptase polymerase chain reaction (RT-PCR) have proven difficult; to date, only one in-frame CBP/MOZ fusion transcript has been reported. We therefore sought other reliable means of detecting CBP rearrangements. We applied fluorescence in situ hybridization (FISH) and Southern blot analyses to a series of AML patients with a t(8;16) and detected DNA rearrangements of both the CBP and the MOZ loci in all cases tested. All six cases examined for CBP rearrangements have breakpoints within a 13 kb breakpoint cluster region at the 5′ end of the CBP gene. Additionally, we used a MOZ cDNA probe to construct a surrounding cosmid contig and detect DNA rearrangements in three t(8;16) cases, all of which display rearrangements within a 6 kb genomic fragment of the MOZ gene. We have thus developed a series of cosmid probes that consistently detect the disruption of the CBP gene in t(8;16) patients. These clones could potentially be used to screen other cancer-associated or congenital translocations involving chromosome band 16p13.3 as well.

Terahertz Imaging of Subjects With Concealed Weapons

In response to the growing interest in developing terahertz imaging systems for concealed weapons detection, the Submillimeter-Wave Technology Laboratory (STL) at the University of Massachusetts Lowell has produced full-body terahertz imagery using coherent active radar measurement techniques. The proof-of-principle results were readily obtained utilizing the compact radar range resources at STL. Two contrasting techniques were used to collect the imagery. Both methods made use of in-house transceivers, consisting of two ultra-stable far-infrared lasers, terahertz heterodyne detection systems, and terahertz anechoic chambers. The first technique involved full beam subject illumination with precision azimuth and elevation control to produce high resolution images via two axis Fourier transforms. Imagery collected in this manner is presented at 1.56THz and 350GHz. The second method utilized a focused spot, moved across the target subject in a high speed two dimensional raster pattern created by a large two-axis positioning mirror. The existing 1.56THz compact radar range was modified to project a focused illumination spot on the target subject several meters away, and receive the back-reflected intensity. The process was repeated across two dimensions, and the resultant image was assembled and displayed utilizing minimal on-the-fly processing. Imagery at 1.56THz of human subjects with concealed weapons are presented and discussed for this scan type.

Terahertz behavior of optical components and common materials

As short range, ground based, surveillance systems operating at terahertz frequencies continue to evolve, increasing attention is being directed towards the behavior of dielectric materials at terahertz frequencies as well as the behavior of optical components used to control terahertz radiation. This work provides an overview of several terahertz optical components such as frequency selective filters, laser output couplers, artificial dielectrics, and electromagnetic absorbers. In addition, a database was established that contains terahertz properties of common materials that have been largely unexplored in this region of the spectrum. The database consists of transmittance and reflectance spectra of a variety of materials measured using Fourier transform infrared spectroscopy techniques from 175 GHz - 2 THz. In addition, ultra-stable, CO2 optically pumped, far-infrared gas lasers were used to collect fixed-frequency transmittance data at 326 GHz, 584 GHz, and 1.04 THz. A Gunn oscillator was used for measurements at 94 GHz.

Terahertz inverse synthetic aperture radar (ISAR) imaging with a quantum cascade laser transmitter

A coherent transceiver using a THz quantum cascade (TQCL) laser as the transmitter and an optically pumped molecular laser as the local oscillator has been used, with a pair of Schottky diode mixers in the receiver and reference channels, to acquire high-resolution images of fully illuminated targets, including scale models and concealed objects. Phase stability of the received signal, sufficient to allow coherent image processing of the rotating target (in azimuth and elevation), was obtained by frequency-locking the TQCL to the free-running, highly stable optically pumped molecular laser. While the range to the target was limited by the available TQCL power (several hundred microwatts) and reasonably strong indoor atmospheric attenuation at 2.408 THz, the coherence length of the TQCL transmitter will allow coherent imaging over distances up to several hundred meters. Image data obtained with the system is presented.

Transformation of the multimode terahertz quantum cascade laser beam into a Gaussian, using a hollow dielectric waveguide

We demonstrate that a short hollow dielectric tube can act as a dielectric waveguide and transform the multimode, highly diverging terahertz quantum cascade laser beam into the lowest order dielectric waveguide hybrid mode, EH(11), which then couples efficiently to the free-space Gaussian mode, TEM(00). This simple approach should enable terahertz quantum cascade lasers to be employed in applications where a spatially coherent beam is required.

Continuous Wave Terahertz Transmission Imaging of Nonmelanoma Skin Cancers

Continuous wave terahertz imaging has the potential to offer a safe, noninvasive medical imaging modality for delineating human skin cancers. Terahertz pulse imaging (TPI) has already shown that there is contrast between basal cell carcinoma and normal skin. Continuous‐wave imaging offers a simpler, lower cost alternative to TPI. The goal of this study was to investigate the feasibility of continuous wave terahertz imaging for delineating skin cancers by demonstrating contrast between cancerous and normal tissue in transmission mode.

Detection of colon cancer by continuous-wave terahertz polarization imaging technique

Abstract. We demonstrate a reflective, continuous-wave terahertz (THz) imaging system to acquire ex vivo images of fresh human colonic excisions. Reflection measurements of 5-mm-thick sections of colorectal tissues were obtained using a polarization-specific detection technique. Two-dimensional THz reflection images of both normal and cancerous colon tissues with a spatial resolution of 0.6 mm were acquired using an optically pumped far-infrared molecular gas laser. Good contrast has been observed between normal and tumorous tissues at 584 GHz frequency. The resulting THz reflection images compared with the tissue histology showed a correlation between cancerous region and increased reflection. We hypothesize that the imaging system and polarization techniques are capable of registering reflectance differences between cancerous and normal colon. However, further investigations are necessary to completely understand the source mechanism behind the contrast and confirm the hypothesis; if true, it likely represents the first continuous-wave THz reflection imaging technique to show sufficient contrast to identify colon tumor margins. Also, it may represent a significant step forward in clinical endoscopic application of THz technology to aid in in vivo colorectal cancer screening.

Frequency stabilization of a single mode terahertz quantum cascade laser to the kilohertz level.

A simple analog locking circuit was shown to stabilize the beat signal between a 2.408 THz quantum cascade laser and a CH(2)DOH THz CO(2) optically pumped molecular laser to 3-4 kHz (FWHM). This is approximately a tenth of the observed long-term (t approximately sec) linewidth of the optically pumped laser showing that the feedback loop corrects for much of the mechanical and acoustic-induced frequency jitter of the gas laser. The achieved stability should be sufficient to enable the use of THz quantum cascade lasers as transmitters in short-range coherent transceivers.

Imaging of ex vivo nonmelanoma skin cancers in the optical and terahertz spectral regions optical and terahertz skin cancers imaging.

We tested the hypothesis that polarization sensitive optical and terahertz imaging may be combined for accurate nonmelanoma skin cancer (NMSC) delineation. Nine NMSC specimens were imaged. 513 μm and 440 nm wavelengths were used for terahertz and optical imaging, respectively. Histopathology was processed for evaluation. Terahertz reflectance of NMSC was quantified. Our results demonstrate that cross-polarized terahertz images correctly identified location of the tumours, whereas cross-polarized and polarization difference optical images accurately presented morphological features. Cross-polarized terahertz images exhibited lower reflectivity values in cancer as compared to normal tissue. Combination of optical and terahertz imaging shows promise for intraoperative delineation of NMSC.