Detection of IgM Antibodies Against a Recombinant Nucleocapsid Protein of Canine Distemper Virus in Dog Sera using a Dot-Blot Assay

A dot-blot assay for the detection of IgM antibodies (ABs) against canine distemper virus (CDV) in canine serum is described. The diagnostic potential of this technique was evaluated by analysing sera from three test groups: (i) specific pathogen-free (SPF) beagle dogs experimentally infected with virulent CDV; (ii) SPF dogs immunized with a combined vaccine containing CDV, and (iii) SPF dogs immunized with a CDV-free vaccine. As antigen for the dot-blot assay we used the recombinant nucleocapsid protein (N protein) of the virulent A75/17 CDV strain. All 12 dogs of group 1, infected with virulent CDV, showed detectable CDV-specific IgM levels in their serum. All dogs of group 2 were also positive for anti-CDV IgM after the first immunization with the CDV-containing vaccine. The four dogs immunized with a CDV-free vaccine (group iii) remained negative throughout the course of the experiment. From these results, we conclude that the IgM detection test, which requires only a single serum sample, is a useful method for diagnosing current or recent CDV infection in CDV-infected or CDV-immunized dogs under experimental conditions.     

The role of bradykinin B1 and B2 receptors for secondary brain damage after traumatic brain injury in mice

Inflammatory mechanisms are known to contribute to the pathophysiology of traumatic brain injury (TBI). Since bradykinin is one of the first mediators activated during inflammation, we investigated the role of bradykinin and its receptors in posttraumatic secondary brain damage. We subjected wild-type (WT), B₁-, and B₂-receptor-knockout mice to controlled cortical impact (CCI) and analyzed tissue bradykinin as well as kinin receptor mRNA and protein expression up to 48 h thereafter. Brain edema, contusion volume, and functional outcome were assessed 24 h and 7 days after CCI. Tissue bradykinin was maximally increased 2 h after trauma (P<0.01 versus sham). Kinin B₁ receptor mRNA was upregulated up to four-fold 24 h after CCI. Immunohistochemistry showed that B₁ and B₂ receptors were expressed in the brain and were significantly upregulated in the traumatic penumbra 1 to 24 h after CCI. B₂R^−/− mice had significantly less brain edema (−51% versus WT, 24 h; P<0.001), smaller contusion volumes (∼50% versus WT 24 h and 7 d after CCI; P<0.05), and better functional outcome 7 days after TBI as compared with WT mice (P<0.05). The present results show that bradykinin and its B₂ receptors play a causal role for brain edema formation and cell death after TBI.     

Assessment of structural lesions in sacroiliac joints enhances diagnostic utility of magnetic resonance imaging in early spondylarthritis

Objective

To compare the diagnostic utility of T1‐weighted and STIR magnetic resonance imaging (MRI) sequences in early spondylarthritis (SpA) using a standardized approach to the evaluation of sacroiliac (SI) joints, and to test whether systematic calibration of readers directed at recognition of abnormalities on T1‐weighted MRI would enhance diagnostic utility.

Methods

Six readers independently assessed T1‐weighted and STIR MRI scans of the SI joints from 187 subjects: 75 ankylosing spondylitis (AS) and 27 preradiographic inflammatory back pain (IBP) patients, and 26 mechanical back pain and 59 healthy volunteer controls ages ≤45 years. The exercise was repeated 6 months later on a random selection of 30 AS patients and 34 controls after calibration directed at lesions visible on T1‐weighted MRI. Specific MRI lesions were recorded according to standardized definitions. In addition to deciding on the presence/absence of SpA, readers were asked which MRI sequence and which type of lesion was the primary basis for their diagnostic conclusion.

Results

Structural lesions were detected in 98% of AS patients and 64% of IBP patients. A diagnosis of SpA was based on T1‐weighted or combined T1‐weighted/STIR sequences in 82% of AS patients and 41% of IBP patients. Calibration enhanced the diagnostic utility of MRI in the majority of readers, especially those considered less experienced; the mean positive and negative likelihood ratios (of 6 readers) were 14.5 and 0.08 precalibration, respectively, and 22.2 and 0.02 postcalibration, respectively.

Conclusion

Recognition of structural lesions on T1‐weighted MRI contributes significantly to its diagnostic utility in early SpA. Rheumatologist training directed at detection of lesions visible on T1‐weighted MRI enhances diagnostic utility.     

Articular cartilage and labral lesions of the glenohumeral joint: diagnostic performance of 3D water-excitation true FISP MR arthrography

Objective  

To evaluate the diagnostic performance of MR arthrography in the detection of articular cartilage and labral lesions of the glenohumeral joint using a transverse 3D water-excitation true fast imaging with steady-state precession (FISP) sequence.     

Myocardial perfusion

Noninvasive cardiac magnetic resonance (CMR) imaging has progressed rapidly over the past few years and will most likely become an integral part of the diagnostic workup of patients with known or suspected coronary artery disease (CAD). In this article the rationale for using perfusion-CMR is discussed, followed by a summary of current state-of-the-art perfusion-CMR techniques that addresses pharmacological stress, monitoring, pulse sequences, and doses of contrast media (CM) for first-pass studies. In the second part, unresolved aspects of perfusion-CMR, such as the lack of fully established and validated imaging protocols, are discussed. The optimum pulse sequence parameters, required cardiac coverage, analysis algorithms, criteria for data quality, and other aspects remain to be defined. Furthermore, since expertise in perfusion-CMR is not yet widely available, training of physicians and technicians to perform perfusion-CMR according to recognized standards is an important future requirement. In the last part of the review, some ideas are proposed to improve the management of patients with known or suspected CAD. This involves making a shift from a "reactive" strategy, in which patients are typically approached when they are symptomatic, to an "active" strategy, in which perfusion-CMR is performed for early detection of high-risk patients so that revascularizations can be performed before potentially deadly infarcts occur. An ideal test for such an active strategy would be highly accurate, reliable, safe (and thus repeatable), and affordable. Large multicenter trials have shown that in experienced centers perfusion-CMR is reliable and repeatable, and it is hoped that future studies will demonstrate its cost-effectiveness as well.     

MRI of meniscal lesions: soft-copy (PACS) and hard-copy evaluation versus reviewer experience

OBJECTIVE: The purpose of our study was to compare diagnostic performance, reviewer confidence, and time requirements in the MRI diagnosis of meniscal tears for three types of reviewers and two types of image documentations (PACS vs hard copies). MATERIALS AND METHODS: An experienced musculoskeletal radiologist (reviewer 1), a fellow in musculoskeletal radiology (reviewer 2), and a junior staff member in orthopedic surgery (reviewer 3) evaluated MR images displayed on PACS monitors and hard copies independently and in a blinded fashion with regard to the presence or absence of meniscal tears. Seventy-one patients (mean age, 45.4 years; range, 16-80 years) were consecutively included if they had undergone both MRI of the knee and arthroscopy within 4 months. Arthroscopy was the standard of reference. Evaluation time and the reviewer's confidence in his or her diagnosis (Visual Analogue Scale, possible values of 0-100) were determined. RESULTS: Accuracies, sensitivities, and specificities in diagnosing meniscal tears were 80-87%, 63-85%, and 87-93% for soft copies and 82-85%, 64-76%, and 87-94.0%, respectively, for hard copies. Intrareviewer differences between PACS and hard copies were not significant for any of the three reviewers (McNemar tests). Reviewer 3 was less sensitive but more specific in the diagnosis of meniscal tears than reviewers 1 and 2. This difference was significant for both the PACS and hard copies. The reviewers' confidence in their diagnoses and evaluation times were not significantly different for PACS and hard copies (analysis of variance with Bonferroni post hoc analysis). CONCLUSION: Differences in the diagnostic performance of suspected meniscal tears depend on reviewer experience rather than on the type of documentation.     

Early gene expression changes by Epstein‐Barr virus infection of B‐cells indicate CDKs and survivin as therapeutic targets for post‐transplant lymphoproliferative diseases

Lymphoproliferative diseases (LPDs) associated with Epstein‐Barr virus (EBV) infection cause significant morbidity and mortality in bone marrow and solid organ transplant recipients. To gain insight into LPD pathogenesis and to identify potential effective therapeutic approaches, we investigated early molecular events leading to B‐cell transformation by gene expression profiling of EBV‐infected B‐cells from tonsils by Affymetrix microarray 72 hr postinfection when the B‐cells hyperproliferation phase starts. Cell cycle and apoptosis were the most significantly affected pathways and enriched gene sets. In particular, we found significantly increased expression of cyclin‐dependent kinase (CDK)1 and CCNB1 (cyclin B1) and of one of their downstream targets BIRC5 (survivin). Importantly, the strong upregulation of the antiapoptotic protein survivin was confirmed in lymphoblastoid cell lines (LCLs) and 71% of EBV‐positive post‐transplant EBV‐LPD lesions scored positive for survivin. The validity of early transforming events for the identification of therapeutic targets for EBV‐LPD was confirmed by the marked antiproliferative effect of the CDK inhibitor flavopiridol on LCLs and by the strong induction of apoptosis by survivin inhibition with YM155 or terameprocol. Our results suggest that targeting of CDKs and/or survivin in post‐transplant EBV‐LPD by specific inhibitors might be an important approach to control and eliminate EBV‐transformed B‐cells that should be further considered.