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Biomedical subjects

A Haase

Publications and source records attributed to A Haase.

At least 19 recordsLinked to original sources

A new in situ hybridization technique for spliced RNA species documents the bone marrow origin of pulmonary macrophages in chronic myelogenous leukemia.

Tissue macrophages derive from monocytes of bone marrow origin. Because monocytes from patients with chronic myelogenous leukemia (CML) contain the Philadelphia chromosome (Ph), it seemed probable that tissue macrophages in CML would originate from the malignant clone. Using powerful molecular techniques, we studied pulmonary alveolar macrophages (PAM) from two patients with CML. PAM from Patient 1, a patient in chronic phase studied before bone marrow transplantation (BMT), contained the Ph by Southern blot analysis. Patient 2, an accelerated phase patient, was studied after post-BMT relapse. PAM from this patient not only contained the Ph, but also expressed the BCR/ABL message documented by a new splice junction in situ hybridization technique. This new technique allows detection of BCR/ABL mRNA and determination of splice useage in individual cells. These data confirm the continued replenishing of PAM from peripheral blood monocytes in non-BMT settings and represent the first direct evidence that tissue macrophages are derived from the malignant clone in patients with CML.

Adult

Scan time reduction in snapshot FLASH MRI.

We describe the speeding up of snapshot FLASH MRI by placing a slice selection experiment before the whole imaging sequence. In conventional snapshot FLASH slice selection is made in every projection, thus consuming a great deal of experimental time. This problem can be overcome by applying only one slice-selective procedure before starting the imaging. The idea is to make a selective saturation of the outer volume and to interrogate the unaffected slice by a train of rapid, hard, low-angle pulses. By this new method the measuring times of snapshot FLASH can be further reduced.

Abdomen

Direct measurement of spin-lattice relaxation times of phosphorus metabolites in human myocardium.

T1 values of phosphorus metabolites visible in human cardiac 31P-MR spectra were determined in 12 volunteers at 1.5 T. Consecutive spectra were acquired with varying pulse repetition time (TR) from 1.6 to 24 s; volume selection was achieved with ISIS. T1's of creatine phosphate (CP), [gamma-P], [alpha-P], and [beta-P]ATP, 2-3 diphosphoglycerate, and phosphodiesters were 6.1 +/- 0.5, 5.4 +/- 0.5, 5.5 +/- 0.5, 5.8 +/- 1.0, 7.6 +/- 1.0, and 5.0 +/- 1.0 s, respectively. CP/ATP ratios showed little change with varying TR; linear regression of CP/ATP vs TR was of borderline significance (r = 0.28, P = 0.06). T1's for CP and ATP were also determined in standard solution (20 mM CP, 10 mM ATP) yielding T1CP of 8.7 +/- 0.2 and T1[gamma-P]-ATP of 9.9 +/- 0.7 s. Thus, T1's for CP and ATP were similar at 1.5 T in both human heart and standard solution. In human cardiac 31P-MR spectra, CP/ATP ratios may need little correction for partial saturation.

2,3-Diphosphoglycerate

Human T-lymphotropic virus type I-associated myelopathy in patients with the acquired immunodeficiency syndrome.

We describe two cases of serologically confirmed human T-lymphotropic virus type I (HTLV-I)-associated myelopathy involving North American men coinfected by the human immunodeficiency virus type 1. Our first patient suffered from a gradually progressive spastic paraparesis for 10 years prior to presenting with Kaposi's sarcoma, while our second patient developed subacutely progressive spastic paraparesis in the setting of full-blown acquired immunodeficiency syndrome. Autopsy examination of the spinal cords from these two cases revealed widespread axonal loss and demyelination principally involving the lateral columns of case no. 1 and the lateral and anterior columns of case no. 2. Vascular sclerosis and hyalinization were prominent in both cases, but in neither was there a conspicuous inflammatory component. In case no. 2, HTLV-I mRNA was not detected by in situ hybridization, but HTLV-I proviral DNA sequences were detected in this case by polymerase chain reaction. Neither case exhibited multinucleated cell (human immunodeficiency virus type 1) myelitis, vacuolar myelopathy, or evidence of HTLV-II infection by polymerase chain reaction assay.

Acquired Immunodeficiency Syndrome

Fast inversion recovery T1 contrast and chemical shift contrast in high-resolution snapshot FLASH MR images.

Fast MR imaging attracts the interest of both clinicians and physicists because new diagnostic information arises with reduced artifacts due to short investigational times. With the acceleration of the Snapshot FLASH MR sequence, the measurement of high-resolution images with 256 x 256 matrix is reported, together with contrasting prepulses that are applied to attain contrast in combination with higher in-plane resolution. Measuring times are in the range of a second. For whole-body imaging, a TR = 5.2 msec and a TE = 2.6 msec could be attained measuring omit 256 x 256 matrix images. Artifact-free images demonstrating T1 contrast and contrast from chemical shift are performed on moving organs (heart, intestine) in different experiments. These applications can easily be performed in a couple of minutes for clinical use. Especially in the lung, short TE and high resolution result in a new imaging quality of pulmonary and mediastinal vessels.

Abdomen

Increased steady-state levels of alpha-fetoprotein mRNA in hepatocellular carcinoma: an analysis by in situ hybridization.

The molecular basis for the augmented production of alpha-fetoprotein is unknown. We have used in situ hybridization of alpha-fetoprotein cDNA to malignant hepatocytes to establish if increased serum alpha-fetoprotein concentrations are related to detectable steady-state levels of alpha-fetoprotein mRNA in hepatocytes. Tumor tissue from four patients with histologically confirmed hepatocellular carcinoma were examined, and the results compared to fetal liver. Northern blot hybridization for alpha-fetoprotein mRNA in tumor tissue was also analyzed. As expected a high number of grains was observed in fetal liver tissue, indicative of high levels of alpha-fetoprotein mRNA physiologically present during pre-natal development. Sections from all patients with high serum concentrations of alpha fetoprotein showed appreciable hybrid formation, which correlated semi-quantitatively with the serum concentrations. However, hybrids were not detected in a patient with a normal serum alpha-fetoprotein. The high alpha-fetoprotein mRNA levels in fetal and neoplastic liver suggest that gene transcription is the mechanism of alpha-fetoprotein production in malignancy, although the control of this mechanism remains speculative.

Adult

A skin MR-marker for radiation treatment planning.

After experiences with fluid filled tubes a gel fitted with an adhesive has been developed, to reproducibly markate treatment ports on the patient's skin. A gel is chosen, that hinders loss of the fluid and smoothly fits to the skin surface due to the adhesive. The clinical use of the gel hopefully will help with the use of NMR image information in radiotherapy treatment planning.

Equipment Design

Snapshot FLASH MRI. Applications to T1, T2, and chemical-shift imaging.

Snapshot FLASH magnetic resonance imaging techniques have been developed to enable real-time imaging of MR parameters. The first realization of the method is based on a 64 X 128 FLASH tomogram acquired within 200 ms, using improved MR system hardware conditions. The soft tissue contrast obtained in FLASH MRI almost disappears by using flip angles of less than 5 degrees and repetition times of 3 ms. This work describes extensions of FLASH MRI placing conventional MR experiments before the whole imaging sequence. This creates images of any desired contrast without changing the measuring time. Examples of inversion-recovery T1, spin-echo T2, chemical-shift-selective, and spectroscopic FLASH MRI are presented. Further extensions to real-time MRI of blood vessels, diffusion coefficients, combination with two-dimensional MR spectroscopy experiments, and other nuclei are discussed.

Humans

3D-snapshot flash NMR imaging of the human heart.

SNAPSHOT-FLASH is a recently developed, ultrafast imaging technique, based on conventional FLASH imaging. The application of this new variant to 3D imaging allows the acquisition of a 128 x 128 x 32 data set in 12.5 seconds without triggering, or for cardiac imaging with gating within 32 heartbeats. Compared to standard 3D-FLASH this is 128 times faster, because triggering is only required when the 3D phase-encoding gradient is incremented. The method depicts for the first time fast three-dimensional views of the human heart without motional artifacts. The images are spin-density weighted. Using suitable prepulses any desired T1- or T2-contrast may be achieved. The generation of 3D movies is possible without an increase of the total scan time.

Heart

Nucleotide sequence analysis of a provirus derived from an individual with tropical spastic paraparesis.

Human T-cell lymphotropic virus type 1 (HTLV-1), the cause of inapparent infections and T-cell leukemias and lymphomas, has also been implicated in two chronic neurological diseases, tropical spastic paraparesis (TSP) and HTLV-1 associated myelopathy (HAM). We initiated a search for a neurotropic variant of HTLV-1 that might be responsible for these chronic progressive myelopathies by cloning and sequencing a provirus from a T-cell line from an individual with TSP. The LTRs and genes of the TSP provirus differ from HTLV-1 by 20-30 nucleotides in each region, but none of the substitutions ostensibly affect functional sites with the exception of the env gene. We document one substitution in the region encoding gp46 common to TSP and HAM proviruses and a mutation that introduces two stop codons in the region encoding gp21. The latter should delete about 100 amino acids from the transmembrane anchor, and, for this reason, the progeny of the sequenced provirus are likely to be defective viruses, maintained in the culture through coinfection of cells with wild-type non-defective HTLV-1. While defective viruses could be responsible for persistent infection of the nervous system in TSP, this cannot be generally the case as we show that HTLV-1 DNA amplified from cell lines from two other individuals with TSP lacked the stop codons. Similarly, comparisons of DNA amplified from HTLV-1 DNA in cases of ATL, HAM, and TSP did not establish a correlation between the mutation in gp46 and neurological disease. The issue of neurotropic variants in HTLV-1 associated neurological disease thus remains an open one which may be resolved in the future by examining proviruses in cells in the lesions in the nervous system; or proviruses in ATL and HAM/TSP which differ in their ability to replicate in glial or neuronal cells.

Amino Acid Sequence

Cardiac and vascular imaging with an MR snapshot technique.

Real-time vascular and cardiac magnetic resonance (MR) imaging has been reported only with echo-planar imaging. In this study, the fast low-angle shot (FLASH) MR imaging sequence was reduced to repetition times of 3 msec and echo times of less than 1.3 msec with use of an improved MR imaging system. The resulting 200-msec MR images (64 X 128 pixels) are called snapshot FLASH images. They allow measurements from dynamic series of MR images depicting processes such as relaxation behavior and the cardiac cycle in the absence of motion and flow artifacts. In animal studies (at 4.7 T) and in studies of human volunteers (at 2.0 T), vascular and cardiac snapshot FLASH images were obtained as a single shot, as reconstructed motion, and as real-time movies. The arbitrary and fast T1 contrast of these images and the reduction of motion artifacts result in favorable applications for the depiction of myocardial and great-vessel anatomy. These clinical applications can be performed on conventional MR imagers with minor technical modifications.

Animals

The 3'-flanking region shared by the human apolipoprotein AI and CIII gene regulates gene expression in cooperation with 5'-flanking elements.

The genes of the apolipoproteins AI and CIII are localized in opposite orientation on chromosome 11 in close vicinity, separated from each other by a non-coding intergenic region of 2.1 kb. The interdependence of their expression has been studied in cotransfection experiments in Hep G2 cells. An expression vector harboring the intergenic region with CAT as reporter gene and likewise pUC 19 derivatives with the putative cis-elements were constructed. Inhibition of gene expression by 5'-flanking elements of the apo AI and CIII gene was observed. Four proteins have been identified in nuclear extracts of Hep G2 cells that bind to several sequences of the intergenic domain but not to the 5'-flanking regions of apo AI or CIII gene. These proteins might be responsible for the silencer effect. The pathway of the regulation of apo CIII and AI expression derived from the experiments described here is supported by mutations in the intergenic region, leading to the phenotype of hypertriglyceridemia, and the stimulatory effect of cholesterol on apo AI transcription in Hep G2 cells.

Apolipoprotein A-I

[Diagnosis of oropharyngeal function with FLASH-MR tomography].

Magnetic resonance imaging (MRI) has several advantages over conventional X-ray methods: the patient is not exposed to radiation; images of any chosen level can be taken without changing the position of the patient; soft tissues are well differentiated; and artefacts due to dental materials are avoided. Thus, in certain fields of ENT diagnosis MRI is superior to computed tomography, for example, in the imaging of acoustic neuromas, glomus tumours and tumours of the parotids, oropharynx and orbit. The measuring time per slice image, which was previously measured in minutes, has been reduced by a factor up to 1000 by the FLASH (fast low angle shot) technique. Thus, it is now possible to follow human physiological processes on an MRI film with a frame speed of 5 pictures using a whole-body magnet. Films of speech, tongue movements and the act of swallowing reveal the value of this technique for the functional diagnosis of disease of the oropharynx. Precise imaging of the anatomical and functional situation, especially of soft tissues, is superior to that of previous methods such as ultrasound, X-ray, and endoscopy.

Deglutition

Variable excitation angle AFP pulses.

RF pulses employing the principle of adiabatic fast passage are finding increasing application with surface coils. A small modification to existing AFP pulses that allows the excitation angle to be chosen largely independently of B1 strength is described here. Rapid 3D imaging applications of such pulses are presented.

Diagnostic Imaging

31P FLASH NMR imaging.

A method of 31P FLASH NMR imaging is described using a low flip angle excitation pulse and three-dimensional Fourier transform spectroscopic imaging (3DFT). In vivo high-resolution 31P NMR spectra are obtained for each of 32 x 32 image elements using a minimum measuring time of 1.5 min. 31P FLASH images of a rat showing the spatial distribution of phosphocreatine, alpha-, beta-, and gamma-ATP have been measured in a 4.7-T magnet with a spatial resolution of 6 mm.

Adenosine Triphosphate