The cda GenoTox assay: a new and sensitive method for detection of environmental genotoxins, including nitroarenes and aromatic amines

A new bacterial test system for detection of genotoxic compounds was developed, based on two new Salmonella typhimurium tester strains, TGO1 and TGO2. Both strains contain a gene fusion between a strong SOS-promotor, P(cda), and the gfp gene, which allows detection of genotoxic compounds that induce the SOS response. SOS induction was detected by means of flow cytometry. TGO1 showed an increased sensitivity to N-methyl-N'-nitro-N-nitrosoguanidine compared with a previously developed strain, which had an Escherichia coli strain as host instead of S. typhimurium. S9 mix was introduced into the assay, making the test system suitable for detection of indirect mutagens. Furthermore, the genes for bacterial nitro-reductase (NR) and o-acetyl transferase (o-AT) were inserted into TGO2, making it an NR- and o-AT-over-expressing strain. This resulted in an assay that was able to detect the nitroarene 1-nitropyrene and the aromatic amine 2-aminoanthracene with high sensitivity.     

Perfusion MRI Derived Indices of Microvascular Shunting and Flow Control Correlate with Tumor Grade and Outcome in Patients with Cerebral Glioma

Objectives

Deficient microvascular blood flow control is thought to cause tumor hypoxia and increase resistance to therapy. In glioma patients, we tested whether perfusion-weighted MRI (PWI) based indices of microvascular flow control provide more information on tumor grade and patient outcome than does the established PWI angiogenesis marker, cerebral blood volume (CBV).

Material and Methods

Seventy-two glioma patients (sixty high-grade, twelve low-grade gliomas) were included. Capillary transit time heterogeneity (CTH) and the coefficient of variation (COV), its ratio to blood mean transit time, provide indices of microvascular flow control and the extent to which oxygen can be extracted by tumor tissue. The ability of these parameters and CBV to differentiate tumor grade were assessed by receiver operating characteristic curves and logistic regression. Their ability to predict time to progression and overall survival was examined by the Cox proportional-hazards regression model, and by survival curves using log-rank tests.

Results

The best prediction of grade (AUC = 0.876; p < 0.05) was achieved by combining knowledge of CBV and CTH in the enhancing tumor and peri-focal edema, and patients with glioblastoma multiforme were identified best by CTH (AUC = 0.763; p<0.001). CTH outperformed CBV and COV in predicting time to progression and survival in all gliomas and in a subgroup consisting of only high-grade gliomas.

Conclusion

Our study confirms the importance of microvascular flow control in tumor growth by demonstrating that determining CTH improves tumor grading and outcome prediction in glioma patients compared to CBV alone.     

Does evaluation of the ligamentous compartment enhance diagnostic utility of sacroiliac joint MRI in axial spondyloarthritis?

IntroductionInflammation of the sacroiliac joints (SIJ) is a fundamental clinical feature of axial spondyloarthritis (SpA). The anatomy of the irregularly shaped SIJ is complex with an antero-inferior cartilaginous compartment containing central hyaline and peripheral fibrocartilage, and a dorso-superior ligamentous compartment. Several scoring modules to systematically assess SIJ magnetic resonance imaging (MRI) in SpA have been developed. Nearly all of them are based on the cartilaginous joint compartment alone. However, there are only limited data about the frequency of inflammatory lesions in the ligamentous compartment and their potential diagnostic utility in axial SpA. We therefore aimed to evaluate the ligamentous compartment on sacroiliac joint MRI for lesion distribution and potential incremental value towards diagnosis of SpA over and above the traditional assessment of the cartilaginous compartment alone.MethodsTwo independent cohorts of 69 and 88 consecutive back pain patients ≤50 years were referred for suspected SpA (cohort A) or acute anterior uveitis plus back pain (cohort B). Patients were classified according to rheumatologist expert opinion based on clinical, radiographic and laboratory examination as having nonradiographic axial SpA (nr-axSpA; n = 51), ankylosing spondylitis (n = 34), or nonspecific back pain (NSBP; n = 72). Five blinded readers assessed SIJ MRI globally for presence/absence of SpA. Bone marrow edema (BME) and fat metaplasia were recorded in the cartilaginous and ligamentous compartment. The incremental value of evaluating the ligamentous additionally to the cartilaginous compartment alone for diagnosis of SpA was graded qualitatively. We determined the lesion distribution between the two compartments, and the impact of the ligamentous compartment evaluation on diagnostic utility.ResultsMRI bone marrow lesions solely in the ligamentous compartment in the absence of lesions in the cartilaginous compartment were reported in just 0–2.0/0–4.0 % (BME/fat metaplasia) of all subjects. Additional assessment of the ligamentous compartment was regarded as essential for diagnosis in 0 and 0.6 %, and as contributory in 28.0 and 7.7 % of nr-axSpA patients in cohorts A and B, respectively. Concomitant BME in both compartments was evident in 11.6–42.0 % of nr-axSpA and 2.1–2.4 % of NSBP patients.ConclusionAssessing the ligamentous compartment on SIJ MRI provided no incremental value for diagnosis of axial SpA. However, concomitant BME in both compartments may help discriminate nr-axSpA from NSBP.     

Recurrent activity in higher order, modality non-specific brain regions: a Granger causality analysis of autobiographic memory retrieval

It has been proposed that the workings of the brain are mainly intrinsically generated recurrent neuronal activity, with sensory inputs as modifiers of such activity in both sensory and higher order modality non-specific regions. This is supported by the demonstration of recurrent neuronal activity in the visual system as a response to visual stimulation. In contrast recurrent activity has never been demonstrated before in higher order modality non-specific regions. Using magneto-encephalography and Granger causality analysis, we tested in a paralimbic network the hypothesis that stimulation may enhance causal recurrent interaction between higher-order, modality non-specific regions. The network includes anterior cingulate/medial prefrontal and posterior cingulate/medial parietal cortices together with pulvinar thalami, a network known to be effective in autobiographic memory retrieval and self-awareness. Autobiographic memory retrieval of previous personal judgments of visually presented words was used as stimuli. It is demonstrated that the prestimulus condition is characterized by causal, recurrent oscillations which are maximal in the lower gamma range. When retrieving previous judgments of visually presented adjectives, this activity is dramatically increased during the stimulus task as ascertained by Granger causality analysis. Our results confirm the hypothesis that stimulation may enhance causal interaction between higher order, modality non-specific brain regions, exemplified in a network of autobiographical memory retrieval.     

Water level fluctuations may decrease phosphate adsorption capacity of the sediment in oligotrophic high mountain lakes

The effect of sediment desiccation and re-wetting on phosphate adsorption and desorption properties was examined in two oligotrophic high mountain lakes (La Caldera and Río Seco, altitude ~3000 m) in the Sierra Nevada National Park, Spain. Decrease in phosphate sorption properties was observed in transects from the littoral zone to dry land (up to the high water mark) in both lakes concomitantly with loss of amorphous oxides of iron and aluminum as revealed by oxalate and dithionite extractions. X-ray diffraction did not indicate increased amounts of crystalline metal oxides, but there was a loss of fine particles (<20 μm) with distance from the shore, probably due to wind erosion. Likely this explains the loss of amorphous metal oxides as they are often enriched in the fine sediment fraction. Changes in P-speciation toward less available pools were also observed as a result of desiccation. When re-wetted under oxic conditions, the sediments, especially those closest to the shore, released phosphate to the overlying water. The loss of adsorption capacity for phosphate upon re-wetting was associated with loss of amorphous iron oxides, and the changes appear to be non-reversible. Thus, both desiccation and re-wetting will lead to a decrease in sediment phosphate sorption capacity and increased water level fluctuations, a possible scenario in climate change, can therefore increase P availability in the water column of these oligotrophic systems.     

Velocity field studies at surgically imposed arterial stenoses on the abdominal aorta in pigs

In order to describe velocity profiles and the size of deterministic and non-deterministic velocity disturbances at arterial stenoses, symmetrical and asymmetrical stenoses with intended area reductions of 50% (‘moderate’) and 85% (‘severe’) were applied on the abdominal aorta in six pigs. Blood velocities were registered by hot-film anemometry in 21 measuring points distributed across the vessel cross-sectional area in one pre-stenotic and three post-stenotic positions. Signal analysis included ensemble averaging, the high-pass filtering technique, and three-dimensional visualization. None of the stenoses affected the pre-stenotic velocity field. Downstream moderate stenoses flow separation and vortex formation were present. Moderate asymmetric stenoses induced turbulence in the post-stenotic velocity field. Immediately downstream of severe stenoses a prominent post-stenotic jet was present. Farther downstream, a multitude of coherent vortices and turbulence dominated the flow field. The transverse distribution of turbulence intensity parallelled with the peak systolic velocity profile, whereas transverse profiles of the relative turbulence intensity (turbulence intensity/mean velocity) revealed peak values in flow field locations with high velocity gradients. Velocity parameters for symmetric and asymmetric severe stenoses were highly comparable. However, the exact degree of stenosis was significantly higher for symmetrical (85%) than for asymmetrical (76%) stenoses. Therefore, recalling that stenosis severity strongly influences the development of velocity disturbances, this indicates that asymmetry of a stenosis is a predictor for blood velocity disturbances.     

Allele-Specific Suppression of Mutant Huntingtin Using Antisense Oligonucleotides: Providing a Therapeutic Option for All Huntington Disease Patients

Huntington disease (HD) is an inherited, fatal neurodegenerative disorder caused by a CAG repeat expansion in the huntingtin gene. The mutant protein causes neuronal dysfunction and degeneration resulting in motor dysfunction, cognitive decline, and psychiatric disturbances. Currently, there is no disease altering treatment, and symptomatic therapy has limited benefit. The pathogenesis of HD is complicated and multiple pathways are compromised. Addressing the problem at its genetic root by suppressing mutant huntingtin expression is a promising therapeutic strategy for HD. We have developed and evaluated antisense oligonucleotides (ASOs) targeting single nucleotide polymorphisms that are significantly enriched on HD alleles (HD-SNPs). We describe our structure-activity relationship studies for ASO design and find that adjusting the SNP position within the gap, chemical modifications of the wings, and shortening the unmodified gap are critical for potent, specific, and well tolerated silencing of mutant huntingtin. Finally, we show that using two distinct ASO drugs targeting the two allelic variants of an HD-SNP could provide a therapeutic option for all persons with HD; allele-specifically for roughly half, and non-specifically for the remainder.     

Magnetic resonance imaging in psoriatic arthritis: a review of the literature

Psoriatic arthritis is a diverse condition that may be characterized by peripheral inflammatory arthritis, axial involvement, dactylitis and enthesitis. Magnetic resonance imaging (MRI) allows visualization of soft tissue, articular and entheseal lesions, and provides a unique picture of the disease process that cannot be gained using other imaging modalities. This review focuses on the literature on MRI in psoriatic arthritis published from 1996 to July 2005. The MRI features discussed include synovitis, tendonitis, dactylitis, bone oedema, bone erosions, soft tissue oedema, spondylitis/sacroiliitis and subclinical arthropathy. Comparisons have been drawn with the more extensive literature describing the MRI features of rheumatoid arthritis and ankylosing spondylitis.