Inactivation of TNF signaling by rationally designed dominant-negative TNF variants

Tumor necrosis factor (TNF) is a key regulator of inflammatory responses and has been implicated in many pathological conditions. We used structure-based design to engineer variant TNF proteins that rapidly form heterotrimers with native TNF to give complexes that neither bind to nor stimulate signaling through TNF receptors. Thus, TNF is inactivated by sequestration. Dominant-negative TNFs represent a possible approach to anti-inflammatory biotherapeutics, and experiments in animal models show that the strategy can attenuate TNF-mediated pathology. Similar rational design could be used to engineer inhibitors of additional TNF superfamily cytokines as well as other multimeric ligands.     

Systemic RNAi in C. elegans requires the putative transmembrane protein SID-1

Double-stranded RNA-mediated gene interference (RNAi) in Caenorhabditis elegans systemically inhibits gene expression throughout the organism. To investigate how gene-specific silencing information is transmitted between cells, we constructed a strain that permits visualization of systemic RNAi. We used this strain to identify systemic RNA interference-deficient (sid) loci required to spread gene-silencing information between tissues but not to initiate or maintain an RNAi response. One of these loci, sid-1, encodes a conserved protein with predicted transmembrane domains. SID-1 is expressed in cells sensitive to RNAi, is localized to the cell periphery, and is required cell-autonomously for systemic RNAi.     

Reacidification Effects on Water Chemistry and Plankton in a Limed Lake in Sweden

Water chemistry and plankton has been monitored in three Lakes in Tyresta National park SE of Stockholm since 1977. Liming operations started in Lake Långsjön and Lake Trehörningen in 1978 and were repeated every 3–5 years, while Lake Årsjön is an unlimed reference Lake. During 1991–1999, the annual pH median in Lake Långsjön and Lake Årsjön ranged between 6.6–7.1 and 5.2–5.8, respectively, and the composition of phyto- and zooplankton in these lakes did not change markedly. After a final treatment in 1991, the liming of Lake Trehörningen was terminated intentionally. As a result, pH decreased from an annual median 7.1 in 1991, to 6.1 in 1999 (5.8 in 1998). Total organic carbon (TOC) did not change markedly during this period, while the levels of calcium decreased. Metals, known to be influenced by acidification, especially cadmium, manganese and aluminium (Al), increased. The labile-inorganic forms of Al also reached higher levels, especially in 1998. Following the decreasing pH, the total number of phytoplankton taxa decreased by ca 40%. Among zooplankton, the cladocerans Holopedium gibberum, Diaphanosoma brachyurum and Daphnia longispina, common during the limed period, became rare.     

Total vertebrectomy for stabilisation of chronic spinal lumbar luxation in a paraplegic dog without nociception

An adult male crossbred dog was referred with a history of a road traffic accident that took place 1 month earlier. Neurological examination revealed paraplegia with absent nociception in the pelvic limbs. On epaxial palpation, significant curvature of the anatomical axis of the spine between the third and fourth lumbar vertebrae was observed, with the presence of a bone end almost piercing the dog's skin. Survey radiographs of the lumbar spine revealed severe dislocation between L3 and L4 vertebrae. During surgery, the spinal cord was not visible between the dislocated segments. Because of difficulties in reducing the lumbar luxation during surgery, vertebrectomy and vertebral shortening were performed. After alignment between vertebrae L3 and L5, eight cortical orthopaedic screws and bone cement were used for fixation. After 30 days, the dog started to use a wheelchair and was considered by its owner to have a good quality of life with no evidence of pain. To the authors’ knowledge, this is the first case of severe luxation treated by total vertebrectomy and spine shortening in a dog. This surgery can be considered as an option in the management of severe spine luxation when the spinal cord is physically transected.     

Expert opinion based modelling of the risk of human infection with H5N1 through the consumption of poultry meat in Germany

According to current scientific opinion the risk of human infection with H5N1 via preparation and consumption of poultry meat is negligible.This opinion has not yet been challenged by a formal risk assessment, due to the lack of empirical data. We have developed a scenario pathway model as a conceptual framework for a formal assessment of the H5N1 risk to humans through consumption of poultry meat and parameterise the model using information derived from expert opinions. The aim of this study was to investigate whether the notion of an overall negligible risk via the oral infection route is consistent with ad hoc data and expert opinions on the relevant parameters of the model.The model is mainly based on expert opinion. A stochastic Monte-Carlo simulation was conducted which took into consideration (amongst others) the exposure and infection of chicken (broiler and layer), turkeys, ducks and geese, the probabilities of detection prior to slaughter, virus survival and contamination during slaughter, as well as during the cutting and preparation of meat in commercial plants and in private households, respectively. The empirical consumption pattern for poultry meat in Germany was taken into account in the simulation.The results show that the risk for the individual consumer is practically zero whereas up to 23 cases per year in Germany might occur if the upper (more pessimistic) ranges of the expert opinions apply.The finding of a low but non-negligible risk to the population is discussed in relation to the epidemiological information available from recent outbreaks in South East Asia.     

Evaluation and treatment of a posttraumatic intrahepatic biloma in a dog

A 9 mo old male mixed-breed dog was presented with a history of chronic vomiting and fever after undergoing a cholecystectomy for the management of traumatic cystic duct rupture associated with biliary effusion 10 days before referral. A 6 cm × 6 cm intrahepatic bile collection, a biloma, was diagnosed on abdominal ultrasound and fine-needle aspiration. The biloma was treated with percutaneous catheter drainage under ultrasonographic guidance. Two years after aspiration, the dog continued to do well.     

Immunocytochemical localization of the Bacillus sphaericus binary toxin components in Culex quinquefasciatus (Diptera: Culicidae) larvae midgut

This study describes the immunocytochemical localization of the Bacillus sphaericus 2362 binary toxin components, BinA and BinB, in Culex quinquefasciatus larvae that had been intoxicated with this entomopathogen. Ultrathin sections of C. quinquefasciatus larvae midgut embedded in the hydrophilic resin L.R. White were incubated with the antibodies anti-BinA or anti-BinB and then revealed with goat anti-rabbit IgG coupled to gold particles. Immunocytochemical detection demonstrated the presence of specific labeling in ultra-thin sections that had been incubated with the BinA antiserum. Gold particles were detected on the apical areas of cell membranes and inside the epithelial cell cytoplasm, particularly the mitochondria, of cells from the gastric caeca and posterior stomach in larvae exposed during 2 or 24 h to the entomopathogen. A similar labeling pattern was observed in ultrathin sections from both midgut regions when incubated with BinB antiserum.     

Electron microscope observations on the structure of membranes from nuclear polyhedral viruses

Electron micrographs show the presence of the polyhedral, the outer, and the inner membranes occurring in nuclear polyhedral bodies fromMalacosoma neustria, Barathra brassicae andAdoxophyes reticulana. The outer membranes appear to be composed of a central, bimolecular leaflet of lipids bounded on either side by carbohydrates and proteins. The polyhedral and inner membranes, however, seem to consist of protein only. The inner membranes are closely attached to the virus rods, and so are the polyhedral membranes to the polyhedral protein.Samenvatting De polyëdermembranen vanMalacosoma neustria, Barathra brassicae enAdoxophyes reticulana, alsmede de buitenste en binnenste membranen om de deeltjes van deze kernpolyëdervirussen, werden met de elektronenmicroscoop bestudeerd (fig. 1A-I, 2A-C). Om de polyëders te fixeren en ter verkrijging van bepaalde contrasten in ultra-dunne coupes werd van verschillende agentia gebruik gemaakt. Vergelijking van de verkregen foto's (fig. 2D-I, 3A-B) wijst uit, dat het buitenste membraan uit een bimoleculaire laag van lipiden bestaat, die aan beide zijden door koolhydraten en eiwitten zijn begrensd (fig. 4); de polyëdermembranen (fig. 3B) bestaan evenals de binnenste membranen echter uitsluitend uit eiwitten. De virusstaafjes en de polyëders zijn respectievelijk door het binnenste membraan en door het polyëdermembraan nauw omsloten.     

A PCR technique for the detection of Jaagsiekte sheep retrovirus in the blood suitable for the screening of ovine pulmonary adenocarcinoma in field conditions

Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring contagious lung neoplasia caused by jaagsiekte sheep retrovirus (JSRV). Although no specific circulating antibodies against the virus can be detected in infected sheep, JSRV proviral DNA sequences can be found in peripheral blood leukocytes (PBLs) in clinically affected and in a proportion of in contact animals. In this study, existing hemi-nested PCR procedure is compared with a new one-step PCR technique that was developed to minimise potential DNA contamination and reduce sample and reagent handling. Different blood preparations were assessed and the best results were achieved on DNA prepared from buffy coat. The sensitivity of this PCR was lower in JSRV infected sheep without lesions of OPA than in clinically affected sheep, which indicate that this PCR may not be not fully appropriate for screening of individual sheep, but rather to provide results at flock level. This PCR is the only currently available blood test for detection of JSRV infected sheep and may be useful in epidemiological studies and in control programmes of OPA.