Temporary disturbance of actin stress fibers in swine kidney cells during pseudorabies virus infection

Rounding and loosening of cells is a consequence of infection with pseudorabies virus (PrV), both in vitro and in vivo. These changes in the normal structure of the cell may be the result of cytoskeletal changes. Immunofluorescence staining of actin filaments and microtubule bundles was performed to examine whether PrV induces a reorganization of these cytoskeletal components in infected swine kidney (SK) cells. Every 2h until 12h post-inoculation (p.i.), cells were washed in cytoskeleton stabilizing buffer (CSB), fixed with paraformaldehyde and washed again with CSB. Cells were permeabilized with a 1/1000 dilution of Triton X-100 and actin filaments were stained by incubating cells with phalloidin-Texas Red. Staining of microtubules was done by incubating the cells subsequently with mouse monoclonal anti-alpha-tubulin and goat anti-mouse IgG-FITC. During the course of infection, actin fibers of SK cells were rearranged in the following sequence: (1) disappearance of thick actin stress fibers between 4 and 6h p.i., (2) complete loss of stress fibers between 6 and 8h p.i., and (3) reappearance of thin stress fibers starting from 10h p.i. In contrast to herpes simplex virus 1 (HSV1) or equine herpesvirus 1 (EHV1), PrV infection did not induce changes in the cellular microtubule network. PrV infection induces a temporary disassembly of actin stress fibers.     

How Can Participatory Forest Management Cooperatives be Successful in Forest Resources Conservation? An Evidence From Ethiopia

ABSTRACT

In many parts of the world, community-based approaches to forest management have been evolving after recognizing that the top-down approach impedes conserving forests effectively. In Ethiopia also, local communities have been involved in participatory forest management (PFM) arrangements for better conservation of forest resources. However, the achievements in improving forest conservation outcomes have been mixed, while some PFMs are successful, others are not. Using data collected from 42 PFM cooperatives from Metema and Quara districts in Amhara dry forest region of Ethiopia, this study seeks to examine the variables that explain the variations in the performances of PFM cooperatives in forest conservation. The study has found that the performance of the PFM cooperatives depends on the key attributes of users, physical resources, and institutions and these attributes explain best for the success of PFMs. Therefore, the study suggests that these attributes along with the external factors like government policy must be given due importance while designing PFM arrangement. Moreover, just to highlight the impact of the external factors, the government policy like the resettlement program must be reconsidered; while the expansion of trade and infrastructure must be encouraged in order to strengthen the performance of PFMs and achieve forest conservation.     

Temporal variation and high-resolution spatial heterogeneity in soil CO2 efflux in a short-rotation tree plantation

Soil CO2 efflux (SR) is the second largest carbon flux on earth. We investigated the driving factors of the seasonal change and short-distance spatial variation in SR in a short-rotation plantation of willow (Salix viminalis Orm). Total annual SR ranged from 723 to 1149 g Cm(-2) year(-1). Both an exponential and a logistic model were fitted to the data, with soil temperature at a depth of 5 cm as the independent variable. The R2 values for individual sampling points ranged from 0.83 to 0.95 and from 0.85 to 0.93 for the exponential and logistic models, respectively, indicating that soil temperature largely determined the seasonal variation in SR. Modeled soil SR at 10 degrees C ranged from 1.22 to 1.95 micromol m(-2) s(-1), whereas modeled annual Q(10) values were between 3.31 and 6.13. These high Q(10) values were attributed to the absence of drought during the study in 2005. When the coefficients of the general SR models were replaced by linear dependencies on soil and vegetation-related characteristics, the resulting spatially explicit exponential and logistic SR models explained 85 and 86%, respectively, of the variability within the dataset. The analysis indicated that soil carbon concentration, leaf area index, soil pH and root biomass caused differences in SR at the short distances considered in this study. However, incorporating information on variables considered to account for spatial variability in the model did not result in a higher R2 compared with a simple temperature function. When the general SR models were applied to independent datasets from the same plantation, the logistic model provided a better fit than the exponential model when drought occurred. Drought greatly reduced the annual Q(10) values of SR.     

Herpesvirus interference with virus-specific antibodies: bridging antibodies, internalizing antibodies, and hiding from antibodies

Herpesviruses have developed different tools to thwart efficient antibody-dependent neutralisation and lysis of virions and elimination of infected cells. This overview will briefly summarize different of these tools, including (i) viral Fc receptors and the resulting process of antibody bridging, (ii) internalization of individual viral proteins and clustered antibody-antigen complexes from the plasma membrane of infected cells, and (iii) directed egress of virus particles to sites of intimate cell-cell contact that are difficult to access for antibodies.     

The current status and future directions of myxoma virus, a master in immune evasion

ABSTRACT: Myxoma virus (MYXV) gained importance throughout the twentieth century because of the use of the highly virulent Standard Laboratory Strain (SLS) by the Australian government in the attempt to control the feral Australian population of Oryctolagus cuniculus (European rabbit) and the subsequent illegal release of MYXV in Europe. In the European rabbit, MYXV causes a disease with an exceedingly high mortality rate, named myxomatosis, which is passively transmitted by biting arthropod vectors. MYXV still has a great impact on European rabbit populations around the world. In contrast, only a single cutaneous lesion, restricted to the point of inoculation, is seen in its natural long-term host, the South-American Sylvilagus brasiliensis and the North-American S. Bachmani. Apart from being detrimental for European rabbits, however, MYXV has also become of interest in human medicine in the last two decades for two reasons. Firstly, due to the strong immune suppressing effects of certain MYXV proteins, several secreted virus-encoded immunomodulators (e.g. Serp-1) are being developed to treat systemic inflammatory syndromes such as cardiovascular disease in humans. Secondly, due to the inherent ability of MYXV to infect a broad spectrum of human cancer cells, the live virus is also being developed as an oncolytic virotherapeutic to treat human cancer. In this review, an update will be given on the current status of MYXV in rabbits as well as its potential in human medicine in the twenty-first century.Table of contentsAbstract1. The virus2. History3. Pathogenesis and disease symptoms4. Immunomodulatory proteins of MYXV4.1. MYXV proteins with anti-apoptotic functions4.1.1. Inhibition of pro-apoptotic molecules4.1.2. Inhibition by protein-protein interactions by ankyrin repeat viral proteins4.1.3. Inhibition of apoptosis by enhancing the degradation of cellular proteins4.1.4. Inhibition of apoptosis by blocking host Protein Kinase R (PKR)4.2. MYXV proteins interfering with leukocyte chemotaxis4.3. MYXV serpins that inhibit cellular pro-inflammatory or pro-apoptotic proteases4.4. MYXV proteins that interfere with leukocyte activation4.5. MYXV proteins with sequence similarity to HIV proteins4.6. MYXV proteins with unknown immune function5. Vaccination strategies against myxomatosis5.1. Current MYXV vaccines5.2. Vaccination campaigns to protect European rabbits in the wild6. Applications of myxoma virus for human medicine6.1. MYXV proteins as therapeutics for allograft vasculopathy and atherosclerosis6.2. Applications for MYXV as a live oncolytic virus to treat cancer7. Discussion and Conclusions8. List of AbbreviationsReferencesAuthor DetailsAuthors' contributionsCompeting interestsFigure LegendsAcknowledgements.     

Evaluation of magnetic resonance imaging for the differentiation of inflammatory,neoplastic, and vascular intradural spinal cord diseases in the dog

Magnetic resonance imaging (MRI) is a common test for dogs with suspected intradural spinal cord lesions, however studies on diagnostic performance for this test are lacking. Objectives of this multi‐institutional, retrospective, case‐control study were to estimate sensitivity and specificity of MRI for (1) distinguishing between histopathologically confirmed intradural spinal cord disease versus degenerative myelopathy in dogs, (2) categorizing intradural spinal cord diseases as neoplastic, inflammatory, or vascular; and (3) determining tumor type within the etiologic category of neoplasia. Additional aims were to (1) determine whether knowledge of clinical data affects sensitivity and specificity of MRI diagnoses; and (2) report interrater agreement for MRI classification of intradural spinal lesions. Cases were recruited from participating hospital databases over a 7‐year period. Three reviewers independently evaluated each MRI study prior to and after provision of clinical information. A total of 87 cases were sampled (17 degenerative myelopathy, 53 neoplasia, nine inflammatory, and eight vascular). Magnetic resonance imaging had excellent (>97.6%) sensitivity for diagnosis of intradural spinal cord lesions but specificity varied before and after provision of clinical data (68.6% vs. 82.4%, P = 0.023). Magnetic resonance imaging had good sensitivity (86.8%) and moderate specificity (64.7–72.5%) for diagnosing neoplasia. Sensitivity was lower for classifying inflammatory lesions but improved with provision of clinical data (48.1% vs. 81.5%, P = 0.015). Magnetic resonance imaging was insensitive for diagnosing vascular lesions (25.0%). Interrater agreement was very good for correctly diagnosing dogs with intradural lesions (? = 0.882–0.833), and good (? = 0.726–0.671) for diagnosing dogs with neoplasia.     

EVALUATION OF THE WARP‐TURBO SPIN ECHO SEQUENCE FOR 3 TESLA MAGNETIC RESONANCE IMAGING OF STIFLE JOINTS IN DOGS WITH STAINLESS STEEL TIBIAL PLATEAU LEVELING OSTEOTOMY IMPLANTS

Susceptibility artifacts caused by ferromagnetic implants compromise magnetic resonance imaging (MRI) of the canine stifle after tibial plateau leveling osteotomy (TPLO) procedures. The WARP‐turbo spin echo sequence is being developed to mitigate artifacts and utilizes slice encoding for metal artifact reduction. The aim of the current study was to evaluate the WARP‐turbo spin echo sequence for imaging post TPLO canine stifle joints. Proton density weighted images of 19 canine cadaver limbs were made post TPLO using a 3 Tesla MRI scanner. Susceptibility artifact sizes were recorded and compared for WARP vs. conventional turbo spin echo sequences. Three evaluators graded depiction quality for the tibial tuberosity, medial and lateral menisci, tibial osteotomy, and caudal cruciate ligament as sufficient or insufficient to make a diagnosis. Artifacts were subjectively smaller and local structures were better depicted in WARP‐turbo spin echo images. Signal void area was also reduced by 75% (sagittal) and 49% (dorsal) in WARP vs. conventional turbo spin echo images. Evaluators were significantly more likely to grade local anatomy depiction as adequate for making a diagnosis in WARP‐turbo spin echo images in the sagittal but not dorsal plane. The proportion of image sets with anatomic structure depiction graded adequate to make a diagnosis ranged from 28 to 68% in sagittal WARP‐turbo spin echo images compared to 0–19% in turbo spin echo images. Findings indicated that the WARP‐turbo spin echo sequence reduces the severity of susceptibility artifacts in canine stifle joints post TPLO. However, variable depiction of local anatomy warrants further refinement of the technique.     

FOCAL SKELETAL MUSCLE UPTAKE OF 99mTECHNETIUM‐HYDROXYMETHYLENE Diphosphonate FOLLOWING PERONEAL NERVE BLOCKS IN HORSES

We have observed focal skeletal muscle uptake of ^99mTechnetium‐hydroxymethylene diphosphonate (Tc‐HDP), which could mimic a tibial lesion, in horses following peroneal nerve blocks. To characterize this observation further, 45 bone phase scintigrams were performed in 12 horses undergoing peroneal nerve blocks. Scans were performed before, and 1, 3, 7, and 14 days postblock. The superficial and deep branches of the peroneal nerve were blocked by injecting 10 ml of 2% mepivacaine in one limb and 20 ml in the other. Images were evaluated for uptake at the block site and uptake likely to mimic a tibial lesion. Regions of interest were placed over the block site and distal tibia. Count density ratios were used to estimate change in uptake intensity over time. The overall proportion affected was 0.52 (95% confidence interval [CI], 0.36–0.68; P<0.001) 1 day postblock and 0.24 (95% CI, 0.13–0.40; P=0.005) 3 days postblock. The overall proportion likely to mimic a tibial lesion was 0.19 (95% CI, 0.09–0.33; P<0.001) 1 day postblock and 0.21 (95% CI, 0.09–0.40; P=0.005) 3 days postblock. Focal skeletal muscle uptake was seen in only one horse 7 days postblock. Increased uptake intensity was associated with higher local anesthetic dose (P=0.042). Peroneal nerve blocks cause focal skeletal muscle uptake of ^99mTc‐HDP on bone phase scintigraphy. This occurs in approximately 50% of blocked limbs and can mimic a tibial lesion on the lateral view in approximately 20% of blocked limbs.     

Equine alphaherpesviruses (EHV-1 and EHV-4) differ in their efficiency to infect mononuclear cells during early steps of infection in nasal mucosal explants

Equine herpesvirus type 1 (EHV-1) replicates extensively in the epithelium of the upper respiratory tract, after which it can spread throughout the body via a cell-associated viremia in mononuclear leukocytes reaching the pregnant uterus and central nervous system. In a previous study, we were able to mimic the in vivo situation in an in vitro respiratory mucosal explant system. A plaquewise spread of EHV-1 was observed in the epithelial cells, whereas in the connective tissue below the basement membrane (BM), EHV-1-infected mononuclear leukocytes were noticed. Equine herpesvirus type 4 (EHV-4), a close relative of EHV-1, can also cause mild respiratory disease, but a cell-associated viremia in leukocytes is scarce and secondary symptoms are rarely observed. Based on this striking difference in pathogenicity, we aimed to evaluate how EHV-4 behaves in equine mucosal explants. Upon inoculation of equine mucosal explants with the EHV-4 strains VLS 829, EQ(1) 012 and V01-3-13, replication of EHV-4 in epithelial cells was evidenced by the presence of viral plaques in the epithelium. Interestingly, EHV-4-infected mononuclear leukocytes in the connective tissue below the BM were extremely rare and were only present for one of the three strains. The inefficient capacity of EHV-4 to infect mononuclear cells explains in part the rarity of EHV-4-induced viremia, and subsequently, the rarity of EHV-4-induced abortion or EHM.