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.     

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.     

Prevalence of feline infectious peritonitis in specific cat breeds

Although known that purebreed cats are more likely to develop feline infectious peritonitis (FIP), previous studies have not examined the prevalence of disease in individual breeds. All cats diagnosed with FIP at a veterinary teaching hospital over a 16-year period were identified. Breed, sex and reproductive status of affected cats were compared to the general cat population and to mixed breed cats evaluated during the same period. As with previous studies sexually intact cats and purebreed cats were significantly more likely to be diagnosed with FIP; males and young cats also had a higher prevalence of disease. Abyssinians, Bengals, Birmans, Himalayans, Ragdolls and Rexes had a significantly higher risk, whereas Burmese, Exotic Shorthairs, Manxes, Persians, Russian Blues and Siamese cats were not at increased risk for development of FIP. Although additional factors doubtlessly influence the relative prevalence of FIP, this study provides additional guidance when prioritizing differentials in ill purebreed cats.     

Outcome of desmoplasty and fasciotomy for desmitis involving the origin of the suspensory ligament in horses: 27 cases (1995-2004)

OBJECTIVE: To determine outcome of percutaneous ultrasound-guided desmoplasty with simultaneous fasciotomy for proximal suspensory desmitis (ie, desmitis of the origin of the suspensory ligament) in horses that have not responded to stall rest. DESIGN: Retrospective case series. ANIMALS: 27 horses. PROCEDURES: Medical records of horses with proximal suspensory desmitis treated by means of desmoplasty with fasciotomy were reviewed. Follow-up information was obtained through telephone conversations with owners and trainers of the horses or by examination of horses at the hospital. RESULTS: 23 of the 27 (85%) horses, including 3 of 4 horses with forelimb lesions and 20 of 23 horses with hind limb lesions, were able to return to full work after surgery and rehabilitation. All horses had ultrasonographic evidence of healing of suspensory ligament lesions. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that desmoplasty with fasciotomy is a viable treatment option in horses with proximal suspensory desmitis that have not responded to stall rest.     

Nocardia infections in cats: a retrospective multi-institutional study of 17 cases

OBJECTIVE: To record 17 cases of nocardiosis in cats from eastern Australia and to compare this series with cases previously reported. DESIGN: Retrospective/prospective study. RESULTS: Nocardia spp infections were diagnosed in 17 cats over 14 years from the three eastern states of Australia. There were no isolates from dogs during this period, but one isolate from a koala and two from dairy cows. The majority of cats presented with spreading lesions of the subcutis and skin associated with draining sinus tract(s). Early cutaneous lesions consisted of circumscribed abscesses. Infections spread at a variable rate, generally by extension to adjacent tissues. Lesions were generally located in regions subjected to cat bite or scratch injuries, including limbs, body wall, inguinal panniculus and nasal bridge. In some other cases, lesions were situated on distal extremities. The clinical course was variable, from chronic, indolent, initially localised infections to acute fulminating disease. Of the 17 cats, 14 were domestic crossbreds and three were purebreds. There was a preponderance of male cats (12 castrated, 1 entire young adult, 1 entire kitten). Nine of 17 cats were 10 years or older. Interestingly, the majority of infections were attributable to N nova. Immediate and/or predisposing causes could be identified in all cases, and included: renal transplantation [one cat]; chronic corticosteroid administration [three cats]; catabolic state following chylothorax surgery [one cat]; fight injuries [seven cats]; FIV infections [three of seven cats tested]. Of the 17 cats, three were apparently cured. Four were thought to be cured, but infection recurred after several months. Three cats responded partially but were euthanased, while another was improving when it died of unrelated complications. Two died despite treatment and two were euthanased without an attempt at therapy. For two cats there were either insufficient records or the patient was lost to follow up. CONCLUSION: Nocardiosis is a rare, serious disease. Currently it is more common in cats than dogs. Nocardial panniculitis may be clinically indistinguishable from the syndrome caused by rapidly growing mycobacteria. Although the prognosis is guarded, patients with localised infections caused by N nova often respond to appropriate therapy. If definitive treatment is delayed because of misdiagnosis, the disease tends to become chronic, extensive and refractory. Insufficient duration of therapy leads to disease recurrence.