Aortic aneurysm and aortic graft infection related to Mycobacterium bovis after intravesical Bacille Calmette-Guérin therapy-a case series.

BACKGROUND: So called "mycotic" aortic aneurysms account for only 0.7 to 1.3% of all aortic aneurysms and are commonly caused by Staphylococcus aureus and Salmonella species. Bacillus Calmette-Guérin (BCG), a live attenuated strain of Mycobacterium bovis, is part of the therapy of non-muscle-invasive bladder cancer (NMIBC). CASE PRESENTATION: We report a case series of three patients with a mycobacterial graft infection related to BCG after surgical treatment of a presumed mycotic aortic aneurysm as an extremely rare complication after NMIBC treatment. All three patients developed aortic aneurysm after BCG instillation and subsequent mycobacterial graft infection. CONCLUSION: Diagnosis requires a high degree of suspicion because of its nonspecific symptoms and imaging. The pathogen is not detected by standard microbiological testing. Treatment includes triple antimycobacterial therapy and radical surgical interventions. Graft preservation may be considered if no anastomosis is involved.

Temporal averaging for analysis of four-dimensional whole-heart computed tomography perfusion of the myocardium: proof-of-concept study.

To assess the feasibility of four-dimensional (4D) whole-heart computed tomography perfusion (CTP) of the myocardium and the added value of temporal averaging of consecutive 3D datasets from different heartbeats for analysis. We included 30 patients with suspected or known coronary artery disease (CAD) who underwent 320-row coronary CT angiography (CTA) and myocardial CTP. Out of these, 15 patients underwent magnetic resonance myocardial perfusion imaging (MR MPI). All CTP examinations were initiated after 3 min of intravenous infusion of adenosine (140 µg/kg/min) and were performed dynamically covering the entire heart every heart beat over a period of 20 ± 3 heart beats. Temporal averaging for dynamic CTP visualisation was analysed for the combination of two, three, four, six, and eight consecutive 3D datasets. Input time attenuation curves (TAC) were delivered from measurement points in the centre of the left ventricle. In all 30 patients, myocardial 4D CTP was feasible and temporal averaging was successfully implemented for all planned combinations of 3D datasets. Temporal averaging of three consecutive 3D datasets showed best performance in the analysis of all CTP image quality parameters: noise, signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), subjective image quality, and diagnostic accuracy with an improvement of SNR and CNR by a factor of 2.2 ± 1.3 and 1.3 ± 0.9. With increasing level of temporal averaging, the input TACs became smoother, but also shorter. Out of the 11 perfusion defects detected with MR MPI, 9 defects were also visible on the 4D CTP images. Whole-heart CTP of the myocardium is feasible and temporal averaging of dynamic datasets improves quantitative image quality parameters and visualization of perfusion defects while further studies are needed to assess its added value for quantification of perfusion parameters.

Patient satisfaction with coronary CT angiography, myocardial CT perfusion, myocardial perfusion MRI, SPECT myocardial perfusion imaging and conventional coronary angiography.

OBJECTIVES: To evaluate patient acceptance of noninvasive imaging tests for detection of coronary artery disease (CAD), including single-photon emission computed tomography myocardial perfusion imaging (SPECT-MPI), stress perfusion magnetic resonance imaging (MRI), coronary CT angiography (CTA) in combination with CT myocardial stress perfusion (CTP), and conventional coronary angiography (CCA). METHODS: Intraindividual comparison of perception of 48 patients from the CORE320 multicentre multinational study who underwent rest and stress SPECT-MPI with a technetium-based tracer, combined CTA and CTP (both with contrast agent, CTP with adenosine), MRI, and CCA. The analysis was performed by using a validated questionnaire. RESULTS: Patients had significantly more concern prior to CCA than before CTA/CTP (p < 0.001). CTA/CTP was also rated as more comfortable than SPECT-MPI (p = 0.001). Overall satisfaction with CT was superior to that of MRI (p = 0.007). More patients preferred CT (46%; p < 0.001) as a future diagnostic test. Regarding combined CTA/CTP, CTP was characterised by higher pain levels and an increased frequency of angina pectoris during the examination (p < 0.001). Subgroup analysis showed a higher degree of pain during SPECT-MPI with adenosine stress compared to physical exercise (p = 0.016). CONCLUSIONS: All noninvasive cardiac imaging tests are well accepted by patients, with CT being the preferred examination. KEY POINTS: • A variety of cardiac imaging tests is available without known patient preference • CTA/CTP shows a lower degree of concern than conventional coronary angiography • CTA/CTP shows higher overall satisfaction compared to stress perfusion magnetic resonance imaging • CTA/CTP is rated as more comfortable than SPECT-MPI • CTA/CTP is the preferred cardiac imaging test.

Effect of butylscopolamine on image quality in MRI of the prostate.

AIM: To evaluate the impact of butylscopolamine on the quality of magnetic resonance imaging (MRI) images of the prostate. MATERIAL AND METHODS: Eighty-two MRI examinations of the prostate were retrospectively analysed. MRI was performed with a combined endorectal/body phased-array coil including proton density-weighted (PD) sequence, T1-weighted turbo spin-echo (TSE)-sequence, and T2-weighted TSE-sequences. Forty milligrams of butylscopolamine was administered intramuscularly in 31 patients (im-group) and intravenously in 30 patients (iv-group). Twenty-one patients did not receive premedication with butylscopolamine (ø-group). Overall image quality, delineation of the bowel wall, and visualization of the prostate, neurovascular bundle, and pelvic lymph nodes were evaluated qualitatively using a five-point scale (from 1=excellent to 5=non-diagnostic/structure not discernible). Motion artefacts within the endorectal coil were quantified by baseline adjusted signal intensities inside the endorectal coil area. RESULTS: Delineation of the bowel wall using the PD-sequence was significantly improved after both intramuscular and intravenous butylscopolamine administration (ø-group: 3.6+/-0.7; im-group: 2.9+/-0.7; iv-group: 2.9+/-0.7; p=0.001). However, there were no significant differences in motion artefacts measured within the endorectal coil (ø-group: 1.18+/-0.14; im-group: 1.15+/-0.11; iv-group: 1.12+/-0.06; p=0.39). There were also no significant differences in qualitative assessment of visualization of the prostate, neurovascular bundle, pelvic lymph nodes, and of overall image quality between the study groups. CONCLUSION: : In conclusion, butylscopolamine had only a small effect on image quality and is not mandatory for MRI of the prostate.

Myosin-V stepping kinetics: a molecular model for processivity.

Myosin-V is a molecular motor that moves processively along its actin track. We have used a feedback-enhanced optical trap to examine the stepping kinetics of this movement. By analyzing the distribution of time periods separating discrete approximately 36-nm mechanical steps, we characterize the number and duration of rate-limiting biochemical transitions preceding each such step. These data show that myosin-V is a tightly coupled motor whose cycle time is limited by ADP release. On the basis of these results, we propose a model for myosin-V processivity.

Myosin-V is a processive actin-based motor.

Class-V myosins, one of 15 known classes of actin-based molecular motors, have been implicated in several forms of organelle transport, perhaps working with microtubule-based motors such as kinesin. Such movements may require a motor with mechanochemical properties distinct from those of myosin-II, which operates in large ensembles to drive high-speed motility as in muscle contraction. Based on its function and biochemistry, it has been suggested that myosin-V may be a processive motor like kinesin. Processivity means that the motor undergoes multiple catalytic cycles and coupled mechanical advances for each diffusional encounter with its track. This allows single motors to support movement of an organelle along its track. Here we provide direct evidence that myosin-V is indeed a processive actin-based motor that can move in large steps approximating the 36-nm pseudo-repeat of the actin filament.

Single-molecule biomechanics with optical methods.

Single-molecule observation and manipulation have come of age. With the advent of optical tweezers and other methods for probing and imaging single molecules, investigators have circumvented the model-dependent extrapolation from ensemble assays that has been the hallmark of classical biochemistry and biophysics. In recent years, there have been important advances in the understanding of how motor proteins work. The range of these technologies has also started to expand into areas such as DNA transcription and protein folding. Here, recent experiments with rotary motors, linear motors, RNA polymerase, and titin are described.

Single molecule force spectroscopy of spectrin repeats: low unfolding forces in helix bundles.

Spectrin repeats fold into triple helical coiled-coils comprising approximately 106 amino acid residues. Using an AFM-related technique we measured the force required to mechanically unfold these repeats to be 25 to 35 pN. Under tension, individual spectrin repeats unfold independently and in an all-or-none process. The dependence of the unfolding forces on the pulling speed reveals that the corresponding unfolding potential is shallow with an estimated width of 1.5 nm. When the unfolded polypeptide strand is relaxed, several domains refold within less than a second. The unfolding forces of the alpha-helical spectrin domains are five to ten times lower than those found in domains with beta-fold, like immunoglobulin or fibronectin Ill domains, where the tertiary structure is stabilized by hydrogen bonds between adjacent strands. This shows that the forces stabilizing the coiled-coil lead to a mechanically much weaker structure than multiple hydrogen-bonded beta-sheets.

Proliferation and motility responses of primary and recurrent gliomas related to changes in epidermal growth factor receptor expression.

Astrocytic neoplasms show a high incidence of elevated or mutated epidermal growth factor receptor (EGFR) expression. Although proliferative effects from EGFR activation are well described, the role that changes in this receptor play in glioma growth and migration remain poorly addressed. This report characterizes changes in the levels of EGFR expression in three glial tumors at initial presentation (resection) and at the time of recurrence. By quantitative flow cytometry the mean level of EGFR expression increased, decreased, or remained the same in different recurrent astrocytomas relative to their primary tumor cells. Immunocytochemistry for EGFR on monolayer cells corroborated the level of expression in the recurrent tumors relative to their matched primary specimen. Immunoprecipitation indicated that 170 kd EGFR was expressed in each of the tumors, and showed normal down regulation following treatment with EGF. Proliferation response to EGF was seen in only 1/6 instances, but was concentration-dependent when observed. Stimulated migration of the cells was frequently seen and was also concentration-dependent on EGF; the magnitude of response was related to the relative level of 170 kd EGFR expression in the cells. EGFR immunostaining of tissue sections from the tumors confirmed the levels of EGFR expressed in primary and recurrent astrocytomas as was seen in the cultured cells. These results indicate that the relative levels of EGFR in early passage cell cultures from glioma specimens concurs with the measured tissue levels of expression. Human glioma cells are more responsive to migration induction than proliferation induction by EGF.

Substrates for astrocytoma invasion.

A better understanding of the influences of specific extracellular substrates, including proteins, glycosaminoglycans, and parenchymal cells, on the invasive behavior of glioma cells would potentially lead to novel forms of treatment aimed at confining the tumor. A monolayer, microliter scale assay was used to investigate how different substrates influenced glioma migration. Basal or unspecific movement (range, 10-260 microns/d) was determined by observing a panel of seven established human glioma cell lines. Migration rates two to five times higher than this basal activity were referred to as preferential and specific glioma migration; these rates generally occurred on merosin and tenascin. Collagen, fibronectin, or vitronectin were less supportive of migration. The glioma cells migrated on hyaluronic acid, but they did not migrate to the extent generally found on the extracellular matrix proteins. Glioma-derived extracellular matrix also served to promote cell migration. This finding implicates a role for either glioma remodeling or synthesis of a permissive environment for local dissemination that may be independent of the constitutive matrix proteins normally found in the brain. Although the glioma cells were able to migrate over monolayers of other glioma cells, they were unable to migrate over astrocytes and fibroblasts. Our findings indicate that the invasive behavior of glioma cells in situ is most likely a consequence of the interplay between the cells' manipulation of the environment and the constitutive ligands associated with specific regions or structures of the brain.

Specific attachment and migration of human astrocytoma cells on human but not murine laminin.

Attachment sites and biological functions of laminin isolated from murine EHS sarcoma have been well studied. Recently several variants of laminin including human placental laminin have been shown to be distinct from EHS-laminin. This study was undertaken to determine attachment, proliferation, and migration phenomena of human astrocytoma cell lines to human and murine sarcoma EHS-laminin. Using short-term attachment assays human placental laminin was shown to be the better substrate for cell adhesion. EHS-laminin mediated approximately 30-50% of the effect observed on human laminin. The astrocytoma cells expressed beta 1, beta 3, and beta 4 subunit mRNA as determined by RT-PCR. Anti-beta 1 antibodies blocked adhesion to EHS-laminin, but antibodies against beta 1, beta 4, and alpha v subunits were all ineffective in blocking adhesion to human laminin. A migration assay showed that astrocytoma cells on human laminin dispersed from a central seeding area, while cells on EHS-laminin remained where they were seeded. The pattern of dispersion could not be accounted for by changes in growth rates of astrocytoma cells on the different proteins, since both cell lines grew equally well on the two laminins. We conclude that unique epitopes on human laminin are recognized by novel receptors on human astrocytoma cells which confer a migratory phenotype to the cells.

The role of extracellular matrix in human astrocytoma migration and proliferation studied in a microliter scale assay.

Ligands in the extracellular matrix (ECM) are known to mediate migration of normal as well as tumor cells via adhesion molecules such as the integrin receptor family. We develop a microliter scale (15-20 microliters total volume) monolayer migration assay to investigate the ability of astrocytoma cells to disperse on surfaces coated with purified human ECM protein ligands. In this system the rate of radial migration of the cell population was constant over time. For human astrocytoma cell lines U-251 and SF-767, laminin and collagen type IV supported a migratory phenotype; fibronectin and vitronectin only minimally supported migration. The different ECM proteins also influenced growth rate: cells on laminin and collagen had a protracted lag phase. Furthermore, migrating cells seeded on laminin or collagen showed a lower labeling index than did stationary cells in the central, crowded region on the same substrate. This micro-scale migration assay should enable detailed molecular and biochemical studies of the determinants of migration.

Determinants of human astrocytoma migration.

A unique characteristic of astrocytic malignancies is their frequent dissemination through the brain. Cellular determinants of migration include adhesion to the substratum, restructuring of the actin cytoskeleton to generate motion, and (in the setting of invasion into tissue) secretion of enzymes for remodeling interstitial space to accommodate forward motion of the migrating cell. In order to better understand these features in the context of local brain invasion by astrocytoma cells, the adhesion and migratory properties of these cells have been investigated in an in vitro monolayer system. Adhesion of 8 different astrocytoma cell lines to different purified human extracellular matrix (ECM) proteins (collagen type IV, cellular fibronectin, laminin, and vitronectin) revealed that there is no "astrocytoma-specific" ECM protein that consistently leads to high cell binding. Similarly, migration of astrocytoma cells was found to be variable and dependent on different ECM proteins. Laminin was frequently the most permissive for adhesion and migration. Adhesion to collagen, fibronectin, and vitronectin was integrin dependent and could be blocked using anti-beta 1 integrin antibodies; in contrast, attachment to laminin could not be blocked using these antibodies. A comparison of adhesion with migration for each of the cell lines on each of the 4 ECM proteins revealed that poor adhesion was associated with minimal migration and that frequently, high adhesion was correlated with rapid migration. When tested for migration on autologous, cell-derived ECM, none of the cell lines were as migratory as they were on one of the purified ECM proteins, with the exception of SF767 cells. Furthermore, it was found that ECM from SF767 cells promoted the migration of other astrocytoma cells. The results from this study indicate that migration is a constitutive behavior of glioma cells which is dependent on, or modified by, the presence or absence of permissive ligands in the environment.

Tumorigenic, invasive, karyotypic, and immunocytochemical characteristics of clonal cell lines derived from a spontaneous canine anaplastic astrocytoma.

Tumor cells from a spontaneously arising canine astrocytoma were isolated and cloned. Three clonally derived cell lines (DL3580 clone 1, DL3580 clone 2, and DL3580 clone 3) were developed and found to express glial fibrillary acidic protein (GFAP) as well as epidermal growth factor receptor (EGFR/c-erbB1). The cell lines were tumorigenic as subcutaneous xenografts or as intracranial implants in athymic mice, or both. Both the monolayer astrocytoma cells and the xenograft tumor cells from clone 2 were aneuploid, with a modal number of 84 chromosomes per metaphase; clones 1 and 3 were also aneuploid with modal numbers of 82 and 75/79, respectively. The histology of both the initial spontaneously occurring tumor in the dog and the intracranial astrocytoma in athymic mice demonstrated features of diffuse infiltration into normal brain. These newly developed canine glioma cell lines are karyotypically stable for 1 yr in culture and carry the same marker chromosomes as the parental lines. These glioma cell lines may serve as models for investigating mechanisms of glioma invasion into brain. Additionally, clonal cell lines with divergent properties isolated from the same tumor may assist in studies of the molecular basis of astrocytoma progression and heterogeneity.