Laboratory validation of a lateral flow device for the detection of CyHV-3 antigens in gill swabs

Cyprinid herpesvirus-3 (CyHV-3) induces the highly contagious koi herpesvirus disease (KHVD) and may result in significant economic losses to the ornamental and food-producing carp industry. Suspicion of KHVD is triggered by clinical signs and confirmed using laboratory techniques. The latter are labour- and time-consuming, require specialised equipment and trained personnel. For rapid, on-site detection of CyHV-3, a lateral flow device (LFD) was developed using two monoclonal antibodies directed towards the viral glycoprotein ORF65. The LFD was highly specific with analytical and diagnostic specificities of 100%. Analytical sensitivity ranged between 1.25 × 10² and 2.40 × 10⁴ plaque forming units per ml for isolates originating from geographically distinct regions. In experimentally infected carp, CyHV-3 was detected as early as 4–5 days post infection. Diagnostic sensitivities of 52.6% and 72.2% relative to PCR were recorded, depending on the viral isolate used. When onset of mortality was taken as reference, diagnostic sensitivities increased to 67.0% and 93.3%. The diagnostic sensitivity for freshly found-dead animals was 100%, irrespective of the virus isolate used. Given the high specificity and ease-of-use for on-site detection of CyHV-3, the LFD was regarded fit for purpose as a first-line diagnostic tool for the identification of acute CyHV-3 infections in KHVD affected (koi) carp.     

Cost‐Effectiveness of Mass Dog Vaccination Campaigns against Rabies in Flores Island,Indonesia

A dynamic deterministic simulation model was developed to determine the cost‐effectiveness of different mass dog vaccination strategies against rabies in a dog population representative of a typical village on Flores Island. Cost‐effectiveness was measured as public cost per averted dog‐rabies case. Simulations started with the introduction of one infectious dog into a susceptible dog population of 399 dogs and subsequently ran for a period of 10 years. The base scenario represented a situation without any control intervention. Evaluated vaccination strategies were as follows: annual vaccination campaigns with short‐acting vaccine (immunity duration of 52 weeks) (AV_52), annual campaigns with long‐acting vaccine (immunity duration of 156 weeks) (AV_156), biannual campaigns with short‐acting vaccine (BV_52) and once‐in‐2‐years campaigns with long‐acting vaccine (O2V_156). The effectiveness of the vaccination strategies was simulated for vaccination coverages of 50% and 70%. Cumulative results were reported for the 10‐year simulation period. The base scenario resulted in three epidemic waves, with a total of 1274 dog‐rabies cases. The public cost of applying AV_52 at a coverage of 50% was US$5342 for a village. This strategy was unfavourable compared to other strategies, as it was costly and ineffective in controlling the epidemic. The costs of AV_52 at a coverage of 70% and AV_156 at a coverage of 70% were, respectively, US$3646 and US$3716, equivalent to US$3.00 and US$3.17 per averted dog‐rabies case. Increasing the coverage of AV_156 from 50% to 70% reduced the number of cases by 7% and reduced the cost by US$1452, resulting in a cost‐effectiveness ratio of US$1.81 per averted dog‐rabies case. This simulation model provides an effective tool to explore the public cost‐effectiveness of mass dog vaccination strategies in Flores Island. Insights obtained from the simulation results are useful for animal health authorities to support decision‐making in rabies‐endemic areas, such as Flores Island.     

Pulmonary embolism following operative treatment of ankle fractures: a report of three cases and review of the literature

The risks of thromboembolism following operative treatment of ankle fractures are deep vein thrombosis (DVT) and pulmonary embolism (PE). These are potentially life-threatening complications. Many orthopedic surgeons fail to appreciate the potential complications of thromboembolic events because of their rare and delayed occurrence in foot and ankle operations. The purpose of this report is to describe the potential for DVT and PE following ankle operations. We present three cases in which patients who underwent operative treatment of ankle fractures subsequently developed PE. We also review the literature on the prevalence of thrombosis, risk factors, methods of prophylaxis, and use of prophylaxis in surgical procedures of the lower extremity.     

Virosome-based active immunization targets soluble amyloid species rather than plaques in a transgenic mouse model of Alzheimer’s disease

Active vaccination with amyloid peptides shows promise for the treatment and prevention of Alzheimer's disease (AD). Several studies in transgenic mouse models of AD have revealed the potency of vaccination to prevent or even clear amyloid plaques from mouse brain. However, the idea that soluble oligomeric species of beta-amyloid (Abeta), rather than plaques, trigger the disease has gained momentum, and current active vaccination strategies affect the levels of total or soluble brain Abeta little or not at all. We describe an active vaccination method based on Abeta1-16 presented on the surface of virosomes, which triggered a dramatic decrease in both soluble Abeta40 (75% reduction; p=0.01) and soluble Abeta42 (62% reduction; p=0.03) in a double transgenic mouse model of AD. Whereas Abeta40 and Abeta42 levels in the insoluble fraction tended to be reduced (by 30% and 27%, respectively), the number of thioflavine-S-positive amyloid plaques was not affected. The high specific antibody responses, obtained without eliciting T-cell reactivity, demonstrate that immunostimulating reconstituted influenza virosomes are a promising antigen carrier system against the neuropathology of AD.     

Failure of medullary tubes. A materials analysis

The objective of this study is to determine the effects of autoclaving on the stress, strain, ultimate tensile strength (UTS), fracture strain, modulus, and yield stress of nylon medullary tubes. There are three reports describing nylon medullary tube failure in the literature. All cases involved the use of nylon medullary tubes during open reduction internal fixation of fractured long bones. We demonstrated increased brittleness and decreased strength with increased exposure of medullary tubes to autoclaving, most dramatically after 100 autoclave cycles. Visual inspection of tubes is a clear indication of material degradation after repeated autoclaving. Furthermore, there is a significant difference in ultimate tensile strength (P < 0.0001) between tubes exposed to less than 100 sterilization cycles compared to tubes exposed to greater than 100 cycles. Likewise, there is a significant decrease in yield stress (P < 0.0004) between the same groups. We recommend disposal and replacement of nylon medullary tubes before they are exposed to 100 autoclaving cycles in order to avoid failure of the device.     

Limited relevance of the CHEK2 gene in hereditary breast cancer

To establish the importance of CHEK2 mutations for familial breast cancer incidence in the German population, we have screened all 14 of the coding exons in 516 families negative for mutations in both the BRCA1 and BRCA2 genes. We found 12 distinct variants in 30 unrelated patients (5.81%), including 5 that are novel and an additional 4 found for the first time in breast cancer. These aberrations were evaluated in 500 healthy women aged over 50 years and in the case of the 2 exon 10 mutations, 1100delC and 1214del4bp, in 1315 randomized healthy controls. According to our results, a statistically significant association for the exon 10 mutations was observed (p = 0.006). The prevalence of the 1100delC mutation in the German population, however, is significantly lower than those reported for other Caucasian populations both in familial breast cancer patients (1.6%) and controls (0.5%), and shows independent segregation with breast cancer in 2 of 4 families analyzed. The remaining 10 variants were more abundant in patients (21) compared to the controls (12) although the difference was not statistically significant. Interestingly, we found no increased breast cancer risk associated with the splice site mutation IVS2+1G-->A or the most common missense mutation I157T, which account for more than half (12/21) of the variants observed in patients. The low prevalence and penetrance of the exon 10 deletion mutations together with no, or an uncertain elevation in risk for other CHEK2 mutations suggests a limited relevance for CHEK2 mutations in familial breast cancer. Further evaluation of the unique variants observed in breast cancer is required to determine if they may play a role in a polygenic model of familial breast cancer. Nevertheless, it seems premature to include CHEK2 screening in genetic testing.     

Cytokines as genetic modifiers in K5–/– mice and in human epidermolysis bullosa simplex

Epidermolysis bullosa simplex (EBS) is a skin disorder caused by fully‐penetrant mutations in the keratin genes KRT5 and KRT14, leading to extensive cytolysis and cell fragility of basal keratinocytes. EBS is subject to environmental conditions and displays high intra‐ and interfamilial variability, suggesting modifying loci. Here, we demonstrate that upregulation of certain cytokines accompanies mutations in keratin 5 (K5) but not in keratin 14 (K14). We find for the first time that cytokines macrophage chemotactic protein (MCP)‐1/[chemokine (C‐C motif) ligand 2] (CCL2), macrophage inflammatory protein (MIP)‐3β/CCL19 and MIP‐3α/CCL20, all regulated by nuclear factor kappa B (NFκB) and involved in the recruitment, maturation, and migration of Langerhans cells (LCs) in the epidermis, are upregulated in the skin of K5^–/–, but not of K14^–/– mice. In neonatal K5^–/– epidermis, the number of LCs was increased two‐fold. At the same time, tumor necrosis factor alpha (TNFα) remained unaltered, demonstrating the specificity of that process. Most remarkably, enhanced LC recruitment within the epidermis was found in five EBS patients carrying mutations in the KRT5 gene but not in EBS patients with KRT14 gene mutations. In agreement with the NFκB‐dependent regulation of these cytokines, we found a decrease in p120‐catenin in the basal epidermis of K5^–/– mice. These data provide the first explanation for distinct, keratin‐type‐specific genotype–phenotype correlations in EBS and represent a rationale to investigate gene loci affecting skin pathology in EBS. Hum Mutat 0, 1–10, 2009. © 2009 Wiley‐Liss, Inc.