Detection of multiple sclerosis lesions in the cervical cord: which of the MAGNIMS 'mandatory' non-gadolinium enhanced sagittal sequences is optimal at 3T?

BACKGROUND AND PURPOSE: The magnetic resonance imaging in multiple sclerosis consensus guidelines currently mandate three sagittal non-contrast enhanced sequences of T2-weighted fast spin echo, proton density-weighted fast spin echo and short tau inversion recovery; however, these particular three sequences have not previously been compared at 3T. This study compared T2-weighted fast spin echo, proton density-weighted fast spin echo, short tau inversion recovery as well as the double inversion recovery sequence for the sagittal detection of multiple sclerosis lesions in the cervical spinal cord at 3T. METHODS: Nineteen multiple sclerosis patients underwent magnetic resonance imaging with 3T sagittal T2-weighted fast spin echo, proton density-weighted fast spin echo, short tau inversion recovery and double inversion recovery between November 2012 and April 2013. Two neuroradiologists independently reviewed the images, and the number of lesions detected on each sequence was recorded. Lesion conspicuity was quantitatively assessed with the lesion-to-cord-contrast ratio and lesion contrast-to-noise ratio. The Wilcoxon signed rank test was performed for statistical analysis. RESULTS: Proton density-weighted fast spin echo and short tau inversion recovery detected 32% more lesions compared to T2-weighted fast spin echo, and 37% more lesions compared to double inversion recovery. The lesion-to-cord-contrast ratio was highest in short tau inversion recovery, while the lesion contrast-to-noise ratio was highest for proton density-weighted fast spin echo. CONCLUSIONS: This study provides the necessary evidentiary support at 3T for the magnetic resonance imaging in multiple sclerosis spinal magnetic resonance imaging protocol consensus guidelines. At 3T sagittal proton density-weighted fast spin echo and short tau inversion recovery sequences allowed improved detection of cervical spinal cord multiple sclerosis lesions, compared to T2-weighted fast spin echo and three-dimensional double inversion recovery magnetic resonance imaging. Utilising T2-weighted fast spin echo alone at 3T is insufficient for lesion detection.

Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9.

The revolutionary RNA-guided endonuclease CRISPR/Cas9 system has proven to be a powerful tool for gene editing in a plethora of organisms. Here, utilizing this system we developed an efficient protocol for the generation of heritable germline mutations in the parasitoid jewel wasp, Nasonia vitripennis, a rising insect model organism for the study of evolution, development of axis pattern formation, venom production, haplo-diploid sex determination, and host-symbiont interactions. To establish CRISPR-directed gene editing in N. vitripennis, we targeted a conserved eye pigmentation gene cinnabar, generating several independent heritable germline mutations in this gene. Briefly, to generate these mutants, we developed a protocol to efficiently collect N. vitripennis eggs from a parasitized flesh fly pupa, Sarcophaga bullata, inject these eggs with Cas9/guide RNA mixtures, and transfer injected eggs back into the host to continue development. We also describe a flow for screening mutants and establishing stable mutant strains through genetic crosses. Overall, our results demonstrate that the CRISPR/Cas9 system is a powerful tool for genome manipulation in N. vitripennis, with strong potential for expansion to target critical genes, thus allowing for the investigation of several important biological phenomena in this organism.

Insulin-mediated activation of the L-arginine nitric oxide pathway in man, and its impairment in diabetes.

AIMS/HYPOTHESIS: Impaired L-arginine transport has been reported in cardiovascular diseases, providing a possible mechanism for reduced nitric oxide (NO) production. Given that cardiovascular diseases are also associated with insulin resistance, and insulin is known to induce vasodilation via a NO-dependent pathway, we hypothesised that abnormal insulin modulation of L-arginine transport may contribute to vascular dysfunction in diabetes. METHODS: Forearm blood flow (FBF) responses to insulin and sodium nitroprusside (SNP) were measured in control and type 2 diabetic volunteers using venous occlusion plethysmography. Effects of intra-arterial insulin on the forearm veno-arterial flux of arginine and related amino acids were determined by HPLC. The effect of locally delivered insulin on arginine transport was assessed during an intra-arterial infusion of [4,5-(3)H] L-arginine. RESULTS: In controls, intrabrachial infusion of 5 mUnits/min insulin lead to a progressive rise in FBF (p<0.001) while this was not evident in diabetics. In support of this observation, we observed a concomitant, significant increase in the flux of N-hydroxy-L-arginine (the NO precursor) in controls (baseline vs. 60 mins insulin: 16.2±12.2 vs. 33.0±13.1 nmol/100 ml tissue/min; p<0.01), whilst no increase was observed in diabetics. Moreover, insulin augmented the clearance of [(3)H]L-arginine from the forearm circulation in controls (baseline vs insulin: 123±22 vs. 150±28 ml/min; p<0.05) but not in diabetics. CONCLUSION: These findings suggest that insulin resistance may contribute substantially to the onset and development of cardiovascular disease in type 2 diabetics via abnormal insulin-mediated regulation of L-arginine transport.