The role of apoptosis in bispecific antibody-mediated T-cell cytotoxicity.

In this report we describe the role of apoptosis in the process of tumour cell killing by bispecific monoclonal antibody (BsMAb)-redirected cytolytic T cells. The BsMAb used, BIS-1, has dual specificity for the CD3 complex on T cells and the pancarcinoma-associated 38 kDa transmembrane antigen EGP-2. BIS-1 allows activated T cells to specifically recognise and kill EGP-2-positive but not EGP-2-negative target cells. An assay was developed to quantify apoptosis in cells by separation of 3H-thymidine-labelled low-molecular, i.e. fragmented, from high-molecular, i.e. non-fragmented DNA. The presence of low molecular weight DNA was measured both within the target cells and in the cell-free supernatant. After exposure to BIS-1-redirected, -activated T cells, apoptosis was observed in EGP-2-positive target cells but not in EGP-2-negative target cells. Also no DNA fragmentation proved to be induced in the activated effector cells during assay. The degree of EGP-2-positive target DNA fragmentation depended on the concentration of BsMAb, the E/T ratio and the incubation time. Using a low E/T ratio (1/1), DNA fragmentation in and 51Cr release from target cells showed similar characteristics and kinetics. At higher E/T ratio (20/1), the 51Cr release from the target cells increased to a greater extent than the percentage fragmented target cell DNA. Inhibitors of DNA fragmentation added to the cytotoxicity assay inhibited not only DNA fragmentation, but also the release of chromium-51 from the target cells, suggesting that apoptosis and cell lysis are closely related in BsMAb-mediated cell killing

Impact of foot-and-mouth disease on mastitis and culling on a large-scale dairy farm in Kenya

Foot and mouth disease (FMD) is a highly transmissible viral infection of cloven hooved animals associated with severe economic losses when introduced into FMD-free countries. Information on the impact of the disease in FMDV-endemic countries is poorly characterised yet essential for the prioritisation of scarce resources for disease control programmes. A FMD (virus serotype SAT2) outbreak on a large-scale dairy farm in Nakuru County, Kenya provided an opportunity to evaluate the impact of FMD on clinical mastitis and culling rate. A cohort approach followed animals over a 12-month period after the commencement of the outbreak. For culling, all animals were included; for mastitis, those over 18 months of age. FMD was recorded in 400/644 cattle over a 29-day period. During the follow-up period 76 animals were culled or died whilst in the over 18 month old cohort 63 developed clinical mastitis. Hazard ratios (HR) were generated using Cox regression accounting for non-proportional hazards by inclusion of time-varying effects. Univariable analysis showed FMD cases were culled sooner but there was no effect on clinical mastitis. After adjusting for possible confounders and inclusion of time-varying effects there was weak evidence to support an effect of FMD on culling (HR = 1.7, 95% confidence intervals [CI] 0.88-3.1, P = 0.12). For mastitis, there was stronger evidence of an increased rate in the first month after the onset of the outbreak (HR = 2.9, 95%CI 0.97-8.9, P = 0.057)

Systemic virus distribution and host responses in brain and intestine of chickens infected with low pathogenic or high pathogenic avian influenza virus

<p>Abstract</p> <p>Background</p> <p>Avian influenza virus (AIV) is classified into two pathotypes, low pathogenic (LP) and high pathogenic (HP), based on virulence in chickens.</p> <p>Differences in pathogenicity between HPAIV and LPAIV might eventually be related to specific characteristics of strains, tissue tropism and host responses.</p> <p>Methods</p> <p>To study differences in disease development between HPAIV and LPAIV, we examined the first appearance and eventual load of viral RNA in multiple organs as well as host responses in brain and intestine of chickens infected with two closely related H7N1 HPAIV or LPAIV strains.</p> <p>Results</p> <p>Both H7N1 HPAIV and LPAIV spread systemically in chickens after a combined intranasal/intratracheal inoculation. In brain, large differences in viral RNA load and host gene expression were found between H7N1 HPAIV and LPAIV infected chickens. Chicken embryo brain cell culture studies revealed that both HPAIV and LPAIV could infect cultivated embryonic brain cells, but in accordance with the absence of the necessary proteases, replication of LPAIV was limited. Furthermore, TUNEL assay indicated apoptosis in brain of HPAIV infected chickens only. In intestine, where endoproteases that cleave HA of LPAIV are available, we found minimal differences in the amount of viral RNA and a large overlap in the transcriptional responses between HPAIV and LPAIV infected chickens. Interestingly, brain and ileum differed clearly in the cellular pathways that were regulated upon an AI infection.</p> <p>Conclusions</p> <p>Although both H7N1 HPAIV and LPAIV RNA was detected in a broad range of tissues beyond the respiratory and gastrointestinal tract, our observations indicate that differences in pathogenicity and mortality between HPAIV and LPAIV could originate from differences in virus replication and the resulting host responses in vital organs like the brain.</p

Quantifying near metal visibility using dual energy computed tomography and iterative metal artifact reduction in a fracture phantom

Purpose: To quantitatively assess CT image quality and fracture visibility using virtual monochromatic imaging and iterative metal artifact reduction (iMAR) in a femoral bone fracture phantom with different fixation implants. Methods: A custom made phantom was scanned at 120-kVp and 140-kVp single-energy and 100/150-kVp dual-energy. Three stainless steel and two titanium implants with different thicknesses were placed on the phantom containing simulated one and two mm fractures. Single-energy CT images were reconstructed with and without iMAR, while DECT images were reconstructed at monochromatic energies between 70 and 190 keV. Non-metal scans were used as a reference. A Fourier power spectrum method and fracture model were used to analyze several anatomical areas. Results: CT-value deviations of titanium implants were much lower compared to stainless steel implants. These deviations decreased for both DECT and iMAR. Fracture visibility, measured with the fracture model, improved the most when DECT was used while artifact reduction benefitted more from iMAR. The optimal monochromatic energy for metal artifact reduction, based on CT-value deviation, varied for each metal between 130 and 150 keV. The fracture model provided a signal-to-noise ratio for the near metal fracture visibility, providing the optimal keV. Conclusion: iMAR and high keV monochromatic images extracted from DECT both reduce metal artifacts caused by different metal fixation implants. Quantitative femoral phantom results show that DECT is superior to iMAR regarding fracture visualization adjacent to metal fixation implants. The introduction of new artifacts when using iMAR impedes its value in near metal fixation implant imaging

Geometric 3D analyses of the foot and ankle using weight-bearing and non weight-bearing cone-beam CT images: The new standard?

Objectives: We hypothesize that three-dimensional (3D) geometric analyses in weight bearing CT-images of the foot and ankle are more reproducible compared to two-dimensional (2D) analyses. Therefore, we compared 2D and 3D analyses on bones of weight-bearing and non weight-bearing cone-beam CT images of healthy volunteers. Methods: Twenty healthy volunteers (10 male, 10 female, mean age 37.5 years) underwent weight-bearing and non weight-bearing cone-beam CT imaging of both feet. Clinically relevant height and angle measurements were performed in 2D and 3D (for example: cuboid height, calcaneal pitch, talo-calcaneal angle, Meary's angle, intermetatarsal angle). Three-dimensional measurements were obtained using automated software. Intra-observer and inter-observer agreement were evaluated for all 2D measurements. Results: Overall intraclass correlation coefficients (ICC's) were higher than 0.750 for most 2D measurements, ranging from 0.352 to 0.995. Calcaneal pitch, angle between the first metatarsal (MT1) and proximal phalange 1, between the fifth metatarsal (MT5) and the calcaneus and heights of the sesamoid bones, navicular, cuboid and talus decreased during weight-bearing in both 2D and 3D results (p < 0.01). Meary's angle was not statistically different in 2D (p = 0.627) and 3D (p = 0.765). Higher coefficients of variation in 2D geometric analysis parameters (0.27 versus 0.16) indicate that 3D analyses are more precise compared to 2D (p < 0.01). Results of left and right feet are comparable for 2D and 3D analyses. Conclusion: Although 2D and 3D geometrical analyses are fundamentally different, automated 3D analyses are more reproducible and precise compared to 2D analyses. In addition, 3D evaluation better demonstrates differences in bone configurations between weight-bearing and non weight-bearing conditions, which may be of value to demonstrate pathology

Improved diagnostic confidence in evaluating bone non-union using virtual monochromatic dual-energy CT

Purpose: The aim of this study was to determine whether virtual monochromatic dual-energy CT imaging improves the evaluation of suspected non-union of the appendicular skeleton treated with titanium or stainless steel intramedullary nails and plates. Method: Forty-one patients with a clinical suspected non-union with hardware in place were included and scanned on a dual-source CT-scanner using 100/Sn150kVp. Images including titanium hardware were extracted at 130 keV. Images including stainless steel hardware were extracted at 150 keV. Monochromatic 70 keV images served as reference. Non-union confirmed during revision surgery was used as gold standard. A musculoskeletal radiologist and orthopedic trauma surgeon evaluated images on image quality, degree and location of consolidation, non-union type and diagnostic confidence. Results: Likert scores with respect to image quality improved from 0.88 to 1.83 (p < 0.001) in high (130 and 150) keV images. High keV images reduced the number of false negative non-unions based on consolidation grade with 5% (p = 0.283). Agreement between observers regarding location of consolidation and non-union type did not improve in 130 and 150 keV images. Diagnostic confidence improved from 1.43 to 2.37 in high keV images compared to 70 keV images (p < 0.001). Overall diagnostic confidence was higher in intramedullary nails than plates (p < 0.05). Conclusions: Use of virtual monochromatic 130 and 150 keV dual-energy CT compared to 70 keV images improves the evaluation of suspected non-union of the appendicular skeleton treated with titanium or stainless steel intramedullary nails and plates

Detection of Bovine Herpesvirus 1 Glycoprotein E Antibodies in Individual Milk Samples by Enzyme-Linked Immunosorbent Assays

The purpose of this study was to determine whether individual milk samples can replace serum samples for the detection of bovine herpesvirus 1 (BHV1) glycoprotein E (gE)-specific antibodies. Serum and milk samples were collected at the same time from cattle in BHV1-free herds, cattle in unvaccinated herds, and cattle in herds that were vaccinated twice with a BHV1 marker vaccine. The samples were tested in two gE enzyme-linked immunosorbent assay (ELISA) systems. In comparison to serum, the results showed that the gE-blocking ELISA was highly sensitive for testing milk samples (0.96). In contrast, the gE ELISA was less sensitive (0.79). The specificities of the gE-blocking ELISA and the gE ELISA for testing milk samples were very high (1.00 and 0.99, respectively). The presented results indicate that individual milk samples, which can be collected relatively easily and inexpensively, can be used instead of individual serum samples in the gE-blocking ELISA for the screening of cattle for BHV1 gE antibodies