Canine parvovirus infection: which diagnostic test for virus?

Five laboratory tests for diagnosis of canine parvovirus type 2 (CPV-2) infection were employed to test 89 faecal samples collected from dogs with diarrhoea. The tests analysed were immunochromatography (IC), haemagglutination (HA), virus isolation (VI), conventional and real-time PCR. IC, HA, VI and conventional or real-time PCR were able, respectively, to detect CPV-2 antigen or nucleic acid in 41, 50, 54, 68 and 73 of the samples. The best correlation was found between conventional and real-time PCR, with an overall agreement of 94.38%. Sixty-eight samples that tested positive by HA, VI or conventional PCR were subjected to antigenic and/or genetic analyses of the CPV-2 strains by monoclonal antibody (MAb), restriction fragment-length polymorphism (RFLP) and/or sequence analyses. In sum, out of the 68 strains analysed, 26 were characterised as CPV-2a, 18 as CPV-2b and 24 as a CPV-2 Glu-426 mutant recently identified in Italy.     

In vivo characterization of the liver fat ¹H MR spectrum

A theoretical triglyceride model was developed for in vivo human liver fat (1) H MRS characterization, using the number of double bonds (-CH=CH-), number of methylene-interrupted double bonds (-CH=CH-CH(2)-CH=CH-) and average fatty acid chain length. Five 3 T, single-voxel, stimulated echo acquisition mode spectra (STEAM) were acquired consecutively at progressively longer TEs in a fat-water emulsion phantom and in 121 human subjects with known or suspected nonalcoholic fatty liver disease. T(2)-corrected peak areas were calculated. Phantom data were used to validate the model. Human data were used in the model to determine the complete liver fat spectrum. In the fat-water emulsion phantom, the spectrum predicted by the model (based on known fatty acid chain distribution) agreed closely with spectroscopic measurement. In human subjects, areas of CH(2) peaks at 2.1 and 1.3 ppm were linearly correlated (slope, 0.172; r = 0.991), as were the 0.9 ppm CH(3) and 1.3 ppm CH(2) peaks (slope, 0.125; r = 0.989). The 2.75 ppm CH(2) peak represented 0.6% of the total fat signal in high-liver-fat subjects. These values predict that 8.6% of the total fat signal overlies the water peak. The triglyceride model can characterize human liver fat spectra. This allows more accurate determination of liver fat fraction from MRI and MRS.     

In vivo EPR: when, how and why?

This special issue is aimed at providing the readers of this journal with an indication of the exciting and important areas in which in vivo electron paramagnetic resonance (EPR) [or equivalently electron spin resonance (ESR)] is making contributions to experimental progress and to provide perspectives on future developments, including the potential for in vivo EPR to be an important new clinical tool. There also are many situations where the combination of in vivo EPR with NMR may be very synergistic. EPR (ESR) is a magnetic resonance-based technique that detects species with unpaired electrons. The technique has become a major tool in diverse fields ranging from biology and chemistry to solid-state physics. In the last few years, many publications have demonstrated that EPR measurements in living animals (in vivo EPR) can provide very significant new insights to physiology, pathophysiology and pharmacology. The most successful applications of in vivo EPR have been non-invasive measurements of oxygen, nitric oxide, bioradicals, pH and redox state, with applications in oncology, cardiology, neuroscience and toxicology. EPR also appears to be the method of choice for measuring radiation dose retrospectively, including the potential to do this in vivo in human subjects. While far from comprehensive, the reviews, original contributions and viewpoints provided in this issue by several leaders in the field of in vivo EPR should provide the readers with confirmation that in vivo EPR is an exciting field that is likely to provide very valuable complementary information for many NMR-based studies in experimental animals and, probably, also for clinical studies.     

Rhinitis: do diagnostic criteria affect the prevalence and treatment?

BACKGROUND: Rhinitis is one of the world's most common health problems. Diagnostic criteria used in community surveys may affect reported prevalence and treatment. METHODS: A proportionately stratified random sample study was performed to investigate the prevalence, comorbidities and management of community-based patients with rhinitis in the tropical urban city of Singapore. RESULTS: The prevalence of at least one, two, three, or four nasal symptoms on most days during the past year in our study population was 25.5%, 13.1%, 6.5%, and 3.0%, respectively. Based on the definition of 'rhinitis' by the International Consensus Report (ICR), the prevalence was 13.1% in Singapore. There was significantly higher prevalence of self-reported allergy, asthma, and common cold/influenza-like illness among the rhinitis group. In the 53% of rhinitis subjects seeking for medical help, 71% visited a primary care physician and 20% an otolaryngologist. Treatments as reported by patients were decongestants (topical or oral) 27%, antibiotics 12%, antihistamines 6%, nasal steroids 3%, surgery 2%, traditional methods 28%, and 22% did not know what medication they had. Subjects considered the effectiveness of treatment unsatisfactory because the majority of them had only partial or no relief with any treatment. CONCLUSIONS: The standardization of the definition of rhinitis in epidemiological studies is of crucial importance, especially when comparing the prevalence between studies. Appropriate patient education by physicians with a good understanding of the nature of rhinitis and the available treatment options (e.g. evidenced-based efficacy, safety, and a good cost-benefit ratio) will maximize patient compliance and treatment outcomes.     

Viral vectors in cancer immunotherapy: which vector for which strategy?

Gene therapy involves the transfer of genetic information to a target cell to facilitate the production of therapeutic proteins and is now a realistic prospect as a cancer treatment. Gene transfer may be achieved through the use of both viral and non-viral delivery methods and the role of this method in the gene therapy of cancer has been demonstrated. Viruses represent an attractive vehicle for cancer gene therapy due to their high efficiency of gene delivery. Many viruses can mediate long term gene expression, while some are also capable of infecting both dividing and non-dividing cells. Given the broadly differing capabilities of various viral vectors, it is imperative that the functionality of the virus meets the requirements of the specific treatment. A number of immunogene therapy strategies have been undertaken, utilising a range of viral vectors, and studies carried out in animal models and patients have demonstrated the therapeutic potential of viral vectors to carry genes to cancer cells and induce anti-tumour immune responses. This review critically discusses the advances in the viral vector mediated delivery of immunostimulatory molecules directly to tumour cells, the use of viral vectors to modify tumour cells, the creation of whole cell vaccines and the direct delivery of tumour antigens in animal models and clinical trials, specifically in the context of the suitability of vector types for specific strategies.     

Telepathology: a diagnostic tool for the millennium?

Many developments in science have their origins in science fiction and telepathology is no exception. The concept was first illustrated in 1924 in the magazine 'Radio News'. It was not until 1980, however, that the first working telepathology system was demonstrated. Although the system was shown to work, it required special hardware, dedicated software and special microwave transmission links to be installed. Little interest was shown worldwide because of the very high cost and the inability of many people to replicate such a system. Ten years later, the personal computer (PC) was able to provide more than adequate performance at low cost for both image display quality and speed, and the development of video technology had resulted in high quality images being produced by television cameras that were now easily affordable. Microscopes were also relatively cheaper. Thus, by 1993 or 1994, all the hardware necessary to produce a telepathology system was available at reasonable cost. Telepathology can now be used for remote primary diagnosis, remote referral to a specialist pathologist, remote teaching, remote presentation of post-mortem or microscopic findings, quality assurance image circulation and feedback, and consensus diagnosis for pathological review in clinical trials. There are two residual problems. The first concerns the speed of data transmission, commonly referred to as the bandwidth. The second is that the software provided by most of the manufacturers and suppliers of these systems is not entirely suitable to the task and the systems are not interoperable. It is clear that the approach of the manufacturers is at present unlikely to produce telepathology systems which pathologists feel comfortable in using. A somewhat different approach is illustrated by the accompanying article in this issue from the Berlin group, where a relatively simple Java-based applet and the Internet are used to allow single or multiple users to view slides on a robotic microscope. This could form the basis for a truly useful system, but still needs modification for some applications.     

Rheumatoid factors: good or bad for you?

BACKGROUND: Rheumatoid factors (RFs) are autoantibodies associated with rheumatoid arthritis. They can be detected in normal individuals, although transiently. This dichotomy has led to questions about the origins and types of RFs. Recently it has been shown that B cells that produce RFs only do so when activated by two signals, one from engagement of the B-cell receptor and the other from recognition of a pathogen-associated molecular pattern through a Toll-like receptor (TLR). These autoantibodies thus link the innate and acquired immune responses. OBJECTIVE: Through a review of the literature, an examination of the current knowledge of RF induction is presented. The focus is on a discussion of a beneficial or detrimental role for RFs in normal individuals and in those with chronic disease. RESULTS: What makes RF 'good' in some cases and 'bad' in others may reflect the type of RF produced. Low-affinity polyreactive IgM RFs are probably beneficial as they aid in the clearance of immune complexes that are more efficiently cleared, and the RF B cell can act as an antigen-presenting cell and stimulate host defense. However, large amounts of high-affinity RFs found in patients with chronic disease may be harmful by participation in a vicious cycle of autoantibody production by stimulation of self lymphocytes, and/or deposition in blood vessels thus causing vasculitis. CONCLUSIONS: Whether RFs are beneficial or detrimental depends on the context in which they are expressed, the type and amount of RF produced, whether the response is perpetuated by TLR ligation and whether other cells are stimulated either directly or indirectly by RF-positive B cells.     

Calcium phosphate cements: competitive drug carriers for the musculoskeletal system?

This paper attempts to provide an insight in the application of calcium phosphate cements (CPC) in the field of drug delivery devices for the musculoskeletal system. Their ability to set once implanted within the body, giving a highly microporous material, allows incorporation of many types of drugs and biologically active molecules, without losing activity and denaturalization. Additionally, by being injectable these materials can be used in the growing market for new technologies of minimally invasive surgery, and in the treatment of difficult accessible sites. All these characteristics, together with the excellent biological behaviour of CPC, make them good candidates for drug delivery devices to be used in the pharmacological treatment of a great number of diseases of the bone tissue.     

Otoscopic examination: A diagnostic contribution for atherosclerotic disease?

Tympanosclerosis is a particular kind of chronic inflammatory response of the middle ear to mechanical injuries and/or infections. This condition is characterised by the formation of plaques presenting ultrastructural analogies with the atherosclerotic lesions, extended to the tympanic membrane and possibly to the ossicular chain. The less severe degree of tympanosclerosis is represented by asymptomatic and clinically unsignificant whitish plaques within the tympanic membrane, detectable at otoscopy. The pathogenesis of this phenomenon is supposed to present a tight relationship with the pathogenesis of atherosclerosis. This observation has been already reported in medical literature, but deserves further clinical confirmations to better define the real extent of the analogies of both affections. A practical implication of this matter of study could be the possibility to find out a fast and non-invasive test as an early marker of an increased risk of atherosclerotic disease: could otoscopy play such a role?     

High-sensitivity epidermal growth factor receptor immunostaining for colorectal carcinomas,compared with EGFR PharmDx?: A study of diagnostic accuracy

Immunostaining for epidermal growth factor receptor (EGFR) is important in the contemporary therapeutic strategy of colorectal carcinomas. We tried to increase detection sensitivity, and compared the high-sensitivity EGFR immunostaining with a worldwide standard, EGFR PharmDx™ (Dako). In order to pursue high-sensitivity EGFR detection, deparaffinized sections were pressure-cooked in 1 mM EDTA solution, pH 8.0. Two mouse monoclonal antibodies against EGFR, clone EGFR2.5 and DAK-H1-WT, and six kinds of secondary detection reagents, including biotin-free catalyzed signal amplification (CSA II), Simple Stain MAX-PO, PolyVue, Novolink, EnVision™ FLEX+, and MACH3, were evaluated to compare the results with those with EGFR PharmDx™, employing a combination of 2-18-C9 as the primary monoclonal antibody and EnVision™ as the secondary reagent. Furthermore, we replaced EnVision™ in the EGFR PharmDx™ kit with CSAII. EGFR detection sensitivity was higher with DAK-H1-WT than with EGFR2.5, and among the secondary reagents, the strongest signals were observed with Novolink. All 30 colorectal carcinomas showed distinct expression of EGFR with our high-sensitivity EGFR immunostaining, while only 16 (53%) gave focal positivity with EGFR PharmDx™. When EnVision™ in EGFR PharmDx™ was replaced by CSA II, strong signals were seen in all cases, and the expression pattern was comparable with our sequence. Non-neoplastic crypt epithelial cells often showed weakly signal with the standard EGFR PharmDx™, but consistently revealed strong membrane staining in the two high-sensitivity sequences. EGFR PharmDx™ frequently gave false negativity. Importantly, EGFR was consistently and sensitively detected when the secondary polymer in the EGFR PharmDx™ kit was simply replaced by CSA II.     

A comparison between simulated and experimental basis sets for assessing short-TE in vivo ¹H MRS data at 1.5 T

A number of algorithms designed to determine metabolite concentrations from in vivo ¹H MRS require a collection of single metabolite spectra, known as a basis set, which can be obtained experimentally or by simulation. It has been assumed that basis sets can be used interchangeably, but no systematic study has investigated the effects of small variations in basis functions on the metabolite values obtained. The aim of this study was to compare the results of simulated with experimental basis sets when used to fit short‐TE ¹H MRS data of variable quality at 1.5 T. Two hundred and twelve paediatric brain tumour spectra were included in the analysis, and each was analysed twice with LCModel? using a simulated and experimental basis set. To determine the influence of data quality on quantification, each spectrum was assessed and 152 were classified as being of ‘good’ quality. Bland–Altman statistics were used to measure the agreement between the two basis sets for all available spectra and only ‘good’‐quality spectra. Monte‐Carlo simulations were performed to investigate the influence of minor shifts in metabolite frequencies on metabolite concentration estimates. All metabolites showed good agreement between the two basis sets, and the average metabolite limits of agreement were approximately ±3.84 mM for all available data and ±0.99 mM for good‐quality data. Errors obtained from the Monte‐Carlo analysis were found to be more accurate than the Cramer–Rao lower bounds (CRLB) for 12 of 15 metabolites when metabolite frequency shifting was considered. For the majority of purposes, a level of agreement of ±0.99 mM between simulated and experimental basis sets is sufficiently small for them to be used interchangeably. Multiple analyses using slightly modified basis sets may be useful in estimating fitting errors, which are not predicted by CRLBs. Copyright © 2010 John Wiley & Sons, Ltd.     

Effect of contrast agent on the results of in vivo 1H MRS of breast tumors – is it clinically significant?

Choline (Cho) signal identification and quantification in (1)H MRS are used in breast cancer diagnosis. However, an influence of the gadolinium-based contrast agent on the Cho amplitude has been reported experimentally. This study aims to identify the impact of gadolinium-based contrast agents on Cho detection and quantification in postcontrast breast MRS. Consecutive patients were recruited prospectively and randomly allocated to two groups. Group A received a neutral (gadolinium diethylenetriaminepentaacetic acid bis-methylamide) and group B an ionic (gadolinium diethylenetriaminepentaacetic acid) contrast agent, each at a dosage of 0.1 mmol/kg. First, the presence of Cho was identified visually. Then, the normalized Cho intensity in malignant lesions was quantified. Multivariate analysis was applied to identify independent influencing factors on Cho. Sixty-three lesions were investigated [A, n = 34; B, n = 29; 43 malignant (one bilaterally malignant), 20 benign]. Cho was identified visually in 14 of 20 malignant tumors in group A and 12 of 22 malignant tumors in group B (p = 0.477). Normalized Cho differed significantly (p = 0.001) between groups A (mean, 26.8 ± 6.0 AU) and B (mean, 18.2 ± 12.5 AU). No linewidth differences were identified (p > 0.05). Multivariate analysis revealed only group membership (A versus B) as an independent predictor of Cho (p = 0.017). The results suggest stronger negative effects of an ionic relative to a neutral gadolinium-based contrast agent on breast tumor MRS in vivo. These results should be considered when conducting and comparing quantitative Cho measurements in the breast.     

Haemorrhaging lesion in the breast: is there a role for embolisation?

Angiosarcoma of the breast is an extremely rare condition. This case illustrates the use of embolisation as a modality of treatment for primary breast angiosarcoma. No other case has been reported on the use of embolisation for this disorder.