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Osteoarthritis (OA) is a degenerative joint disease for which there are no disease-modifying drugs. It is a leading cause of disability in the UK. Increasing age and obesity are both major risk factors for OA and the health and economic burden of this disease will increase in the future. Focusing on compounds from the habitual diet that may prevent the onset or slow the progression of OA is a strategy that has been under-investigated to date. An approach that relies on dietary modification is clearly attractive in terms of risk/benefit and more likely to be implementable at the population level. However, before undertaking a full clinical trial to examine potential efficacy, detailed molecular studies are required in order to optimise the design. This review focuses on potential dietary factors that may reduce the risk or progression of OA, including micronutrients, fatty acids, flavonoids and other phytochemicals. It therefore ignores data coming from classical inflammatory arthritides and nutraceuticals such as glucosamine and chondroitin. In conclusion, diet offers a route by which the health of the joint can be protected and OA incidence or progression decreased. In a chronic disease, with risk factors increasing in the population and with no pharmaceutical cure, an understanding of this will be crucial.

Objective: Sulforaphane (SFN) has been reported to regulate signaling pathways relevant to chronic diseases. The aim of this study was to investigate the impact of SFN treatment on signaling pathways in chondrocytes and to determine whether sulforaphane could block cartilage destruction in osteoarthritis., Methods: Gene expression, histone acetylation, and signaling of the transcription factors NF-E2-related factor 2 (Nrf2) and NF-κB were examined in vitro. The bovine nasal cartilage explant model and the destabilization of the medial meniscus (DMM) model of osteoarthritis in the mouse were used to assess chondroprotection at the tissue and whole-animal levels., Results: SFN inhibited cytokine-induced metalloproteinase expression in primary human articular chondrocytes and in fibroblast-like synovial cells. SFN acted independently of Nrf2 and histone deacetylase activity to regulate metalloproteinase expression in human articular chondrocytes but did mediate prolonged activation of JNK and p38 MAPK. SFN attenuated NF-κB signaling at least through inhibition of DNA binding in human articular chondrocytes, with decreased expression of several NF-κB-dependent genes. Compared with cytokines alone, SFN (10 μM) abrogated cytokine-induced destruction of bovine nasal cartilage at both the proteoglycan and collagen breakdown levels. An SFN-rich diet (3 μmoles/day SFN versus control chow) decreased the arthritis score in the DMM model of osteoarthritis in the mouse, with a concurrent block of early DMM-induced gene expression changes., Conclusion: SFN inhibits the expression of key metalloproteinases implicated in osteoarthritis, independently of Nrf2, and blocks inflammation at the level of NF-κB to protect against cartilage destruction in vitro and in vivo., (© The Authors. Arthritis & Rheumatism is published by Wiley Periodicals, Inc. on behalf of the American College of Rheumatology.)

In autumn 2011, 11 illegally imported animals were seized from a farm in southern Norway. These included four raccoon dogs (Nyctereutes procyonoides), four raccoons (Procyon lotor), and three South American coatis (Nasua nasua), all considered alien species in Norway. An additional two raccoons had escaped from the farm prior to seizure. The seized animals were euthanized and postmortem examination revealed that the four raccoons had moderate to high numbers of the zoonotic nematode Baylisascaris procyonis in their intestines, ranging from 11 to 115 nematodes per small intestine, with a mean of 53. The identity of the nematodes was confirmed using molecular analysis of ITS-1, ITS-2, cytochrome C oxidase 1, and 18S. Echinococcus multilocularis was not detected in any of the 11 animals. Toxocara and Toxascaris sp. eggs were detected in the feces of two raccoons, and two coatis had coccidia oocysts (80 and 360 oocysts per gram). Domestic dogs and other wildlife on the farm had potential access to the animal pens. Given that the eggs can remain infective for years in the environment, local veterinary and health authorities will need to remain vigilant for symptoms relating to infection with B. procyonis.

Prevalence, abundance and instar composition of Ixodes ricinus as found on one ear collected from 1019 moose (Alces alces), red deer (Cervus elaphus) and roe deer (Capreolus capreolus), shot during hunting (August-December) 2001-2003, are reported. The animals originated from 15 coastal municipalities (CM), seven municipalities bordering to coastal municipalities (BCM) and four inland municipalities (IM), in Norway, between latitudes 58-66° N. I. ricinus occurred endemically in all CM and BCM up to 63°30' N, whereas it was non-endemic further north and in the IM. This geographical distribution of the tick along the coast of southern Norway was largely in accordance with that reported as far back as the 1940s. Our results therefore did not indicate any large scale northwards expansion of I. ricinus in Norway during the 60 year-period between the two studies. However, the prevalence of infestation and tick abundance were significantly higher in CM as compared to BCM. The prevalence and abundance by month were highest during August and September, gradually decreasing towards December. The considerable prevalence of ticks in November, as well as findings in December, would seem to indicate a prolonged tick season as compared with the studies carried out 60 years ago. A total of 8920 ticks were isolated from 439 of the 603 animals examined in endemic municipalities, and the maximum number of ticks found on one single ear was 204. Attached adult ticks were primarily found among the long hairs at base of the ear, whereas nymphs and larvae were seen all over the outer surface of the pinna, for larvae especially at the edge and tip of the ear. Nymphs were the dominant instar, constituting 74% of the total tick count. The proportion of larvae and adult ticks was 13% and 12%. A significantly higher proportion of adult ticks and lower proportion of immature stages were found in moose, as compared to red deer and roe deer. The same apparently size-associated preference of adult ticks was also found for adult animals (all species) as compared to calves. Other grossly detected ectoparasites included the lice Solenopotes burmeisteri in red deer and Damalinia meyeri in roe deer, and the deer ked fly, Lipoptena cervi, in moose and roe deer. This is believed to be the first systematic study on the instar composition by I. ricinus infestation in free-ranging cervids. The examination of ears from hunted cervids should be recognized as a rational way of obtaining data on the geographical distribution and abundance of this tick in nature., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Objective: To examine the ability of a broad-spectrum histone deacetylase (HDAC) inhibitor to protect cartilage in vivo, and to explore the effects of class-selective HDAC inhibitors and small interfering RNA (siRNA)-induced knockdown of HDACs on metalloproteinase expression and cartilage degradation in vitro., Methods: A destabilization of the medial meniscus (DMM) model was used to assess the in vivo activity of the HDAC inhibitor trichostatin A (TSA). Human articular chondrocytes (HACs) and SW-1353 chondrosarcoma cells were treated with cytokines and TSA, valproic acid, MS-275, or siRNA, and quantitative reverse transcription-polymerase chain reaction was performed to determine the effect of treatment on metalloproteinase expression. HDAC inhibitor activity was detected by Western blotting. A bovine nasal cartilage (BNC) explant assay was performed to measure cartilage resorption in vitro., Results: Systemically administered TSA protected cartilage in the DMM model. TSA, valproic acid, and MS-275 repressed cytokine-induced MMP1 and MMP13 expression in HACs. Knockdown of each class I HDAC diminished interleukin-1-induced MMP13 expression. All of the HDAC inhibitors prevented degradation of BNC, in which TSA and MS-275 repressed cytokine-induced MMP expression., Conclusion: Inhibition of class I HDACs (HDAC-1, HDAC-2, HDAC-3) by MS-275 or by specific depletion of HDACs is capable of repressing cytokine-induced metalloproteinase expression in cartilage cells and BNC explants, resulting in inhibition of cartilage resorption. These observations indicate that specific inhibition of class I HDACs is a possible therapeutic strategy in the arthritides., (Copyright © 2013 by the American College of Rheumatology.)

Inter-country travel of companion animals provides an opportunity for introduction of zoonotic pathogens, such as rabies virus and Echinococcus spp. Regulations are in place to control this threat, but Schengen Agreements mean that border controls between some European countries are minimal, and animals may enter countries without any checks that they have been appropriately treated. Veterinarians provide an important source of information for people intending to travel with their pets. We conducted a telephone survey to investigate provision of correct advice to someone intending to travel with their dog to Norway. Mainland Norway is considered free of both rabies and E. multilocularis and is a signatory to the Schengen Agreement. Ten randomly selected veterinary clinics were surveyed in Austria, Belgium (Wallonia), Finland, France, Germany, Norway, Sweden, Switzerland and United Kingdom. The information provided was scored as correct, incorrect or incomplete. The information provided by secondary information sources (website or government agency), which the clinic had referred the caller to, was also assessed (correct, incorrect, incomplete). Whilst the majority of clinics provided appropriate information regarding rabies, many clinics did not provide correct information regarding treatment for E. multilocularis. Less than one in 10 clinics provided the correct information regarding both pathogens directly at the time of calling. The correct information was obtained, once taking into account secondary sources, just 62% of the time. Countrywise, most clinics in Finland provided correct advice, either directly or indirectly via referring the caller to another source, whilst the majority in Belgium, Germany and France did not. The apparent paucity of readily accessible, correct advice for owners intending to travel with their dogs is concerning. The compulsory treatment regulations are only as good as the checks that ensure compliance, and this is also lacking in some countries., (2012 Blackwell Verlag GmbH)

Routine Trichinella meat inspection at the slaughterhouse detected one larva in a pooled batch of 100 pig samples. The larva was sent to the Norwegian Veterinary Institute (NVI) for species identification.Morphological examination revealed that the larva was not Trichinella spp. Molecular analysis was performed. PCR and sequencing of 5S/ITS identified the larva as Toxocara cati. A second round of digests was carried out at the meat inspection laboratory, in smaller batches to try to identify the infected animal. No further larvae were detected and it was not possible to identify which of the 100 animals the larva had come from. This is the first time that Toxocara cati has been reported in slaughterhouse pigs in Norway.Although the infected individual could not be identified, the meat originated from one of six potential farms. A small survey regarding rodent control and cats was sent to each of these farms. Cats had restricted access to food storage areas (two farms reported that cats had access) whilst none of the farms allowed cats into the production housing. Cats were, however, present on all the farms (mostly stray cats of unknown health status). Half of the farms also reported seeing rodents in the pig housing during the previous six months and half reported finding rodents in the feed and straw storage areas. We were unable to narrow down the source of infection - however contamination of food or bedding material, with cat faeces or infected rodents, in addition to the presence of infected rodents in pig housing remain potential routes of infection.

In the past three decades, Echinococcus multilocularis, the cause of human alveolar echinococcosis, has been reported in several new countries both in definitive hosts (canids) as well as in people. Unless treated, infection with this cestode in people is fatal. In previously endemic countries throughout the Northern Hemisphere, geographic ranges and human and animal prevalence levels seem to be increasing. Anthropogenic influences, including increased globalisation of animals and animal products, and altered human/animal interfaces are thought to play a vital role in the global emergence of this pathogenic cestode. Molecular epidemiological techniques are a useful tool for detecting and tracing introductions, and differentiating these from range expansions., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Dupuytren's disease (DD) is a common fibrotic condition of the palmar fascia, leading to deposition of collagen-rich cords and progressive flexion of the fingers. The molecular mechanisms underlying the disease are poorly understood. We have previously shown altered expression of extracellular matrix-degrading proteases (matrix metalloproteases, MMPs, and 'a disintegrin and metalloprotease domain with thrombospondin motifs', ADAMTS, proteases) in palmar fascia from DD patients compared to control and shown that the expression of a sub-set of these genes correlates with post-operative outcome. In the current study we used an in vitro model of collagen contraction to identify the specific proteases which mediate this effect. We measured the expression of all MMPs, ADAMTSs and their inhibitors in fibroblasts derived from the palmar fascia of DD patients, both in monolayer culture and in the fibroblast-populated collagen lattice (FPCL) model of cell-mediated contraction. Key proteases, previously identified in our tissue studies, were expressed in vitro and regulated by tension in the FPCL, including MMP1, 2, 3, 13 and 14. Knockdown of MMP2 and MMP14 (but not MMP1, 3 and 13) inhibited cell-mediated contraction, and knockdown of MMP14 inhibited proMMP-2 activation. Interestingly, whilst collagen is degraded during the FPCL assay, this is not altered upon knockdown of any of the proteases examined. We conclude that MMP-14 (via its ability to activate proMMP-2) and MMP-2 are key proteases in collagen contraction mediated by fibroblasts in DD patients. These proteases may be drug targets or act as biomarkers for disease progression., (© 2012 Elsevier B.V. All rights reserved.)

Background: The fox tapeworm Echinococcus multilocularis has foxes and other canids as definitive host and rodents as intermediate hosts. However, most mammals can be accidental intermediate hosts and the larval stage may cause serious disease in humans. The parasite has never been detected in Sweden, Finland and mainland Norway. All three countries require currently an anthelminthic treatment for dogs and cats prior to entry in order to prevent introduction of the parasite. Documentation of freedom from E. multilocularis is necessary for justification of the present import requirements., Methods: The probability that Sweden, Finland and mainland Norway were free from E. multilocularis and the sensitivity of the surveillance systems were estimated using scenario trees. Surveillance data from five animal species were included in the study: red fox (Vulpes vulpes), raccoon dog (Nyctereutes procyonoides), domestic pig, wild boar (Sus scrofa) and voles and lemmings (Arvicolinae)., Results: The cumulative probability of freedom from EM in December 2009 was high in all three countries, 0.98 (95% CI 0.96-0.99) in Finland and 0.99 (0.97-0.995) in Sweden and 0.98 (0.95-0.99) in Norway., Conclusions: Results from the model confirm that there is a high probability that in 2009 the countries were free from E. multilocularis. The sensitivity analyses showed that the choice of the design prevalences in different infected populations was influential. Therefore more knowledge on expected prevalences for E. multilocularis in infected populations of different species is desirable to reduce residual uncertainty of the results.

Background: Oxidative stress is proposed as an important factor in osteoarthritis (OA)., Objective: To investigate the expression of the three superoxide dismutase (SOD) antioxidant enzymes in OA., Methods: SOD expression was determined by real-time PCR and immunohistochemistry using human femoral head cartilage. SOD2 expression in Dunkin-Hartley guinea pig knee articular cartilage was determined by immunohistochemistry. The DNA methylation status of the SOD2 promoter was determined using bisulphite sequencing. RNA interference was used to determine the consequence of SOD2 depletion on the levels of reactive oxygen species (ROS) using MitoSOX and collagenases, matrix metalloproteinase 1 (MMP-1) and MMP-13, gene expression., Results: All three SOD were abundantly expressed in human cartilage but were markedly downregulated in end-stage OA cartilage, especially SOD2. In the Dunkin-Hartley guinea pig spontaneous OA model, SOD2 expression was decreased in the medial tibial condyle cartilage before, and after, the development of OA-like lesions. The SOD2 promoter had significant DNA methylation alterations in OA cartilage. Depletion of SOD2 in chondrocytes increased ROS but decreased collagenase expression., Conclusion: This is the first comprehensive expression profile of all SOD genes in cartilage and, importantly, using an animal model, it has been shown that a reduction in SOD2 is associated with the earliest stages of OA. A decrease in SOD2 was found to be associated with an increase in ROS but a reduction of collagenase gene expression, demonstrating the complexities of ROS function.

Objective: Increasing evidence implicates serine proteinases in pathologic tissue turnover. The aim of this study was to assess the role of the transmembrane serine proteinase matriptase in cartilage destruction in osteoarthritis (OA)., Methods: Serine proteinase gene expression in femoral head cartilage obtained from either patients with hip OA or patients with fracture to the neck of the femur (NOF) was assessed using a low-density array. The effect of matriptase on collagen breakdown was determined in cartilage degradation models, while the effect on matrix metalloproteinase (MMP) expression was analyzed by real-time polymerase chain reaction. ProMMP processing was determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis/N-terminal sequencing, while its ability to activate proteinase-activated receptor 2 (PAR-2) was determined using a synovial perfusion assay in mice., Results: Matriptase gene expression was significantly elevated in OA cartilage compared with NOF cartilage, and matriptase was immunolocalized to OA chondrocytes. We showed that matriptase activated proMMP-1 and processed proMMP-3 to its fully active form. Exogenous matriptase significantly enhanced cytokine-stimulated cartilage collagenolysis, while matriptase alone caused significant collagenolysis from OA cartilage, which was metalloproteinase-dependent. Matriptase also induced MMP-1, MMP-3, and MMP-13 gene expression. Synovial perfusion data confirmed that matriptase activates PAR-2, and we demonstrated that matriptase-dependent enhancement of collagenolysis from OA cartilage is blocked by PAR-2 inhibition., Conclusion: Elevated matriptase expression in OA and the ability of matriptase to activate selective proMMPs as well as induce collagenase expression make this serine proteinase a key initiator and inducer of cartilage destruction in OA. We propose that the indirect effects of matriptase are mediated by PAR-2, and a more detailed understanding of these mechanisms may highlight important new therapeutic targets for OA treatment.

The cardiovascular nematode Angiostrongylus vasorum is spreading in the fox and dog populations of northern Europe. A. vasorum can result in severe clinical manifestations in dogs; therefore, specific diagnosis is crucial for assessing its prevalence. In the present study, faecal samples from foxes and domestic dogs were tested by a new polymerase chain reaction (PCR) targeting the second internal transcribed region of the ribosomal DNA (ITS2) of A. vasorum. Initial isolation of faecal larvae by sieving facilitated the processing of larger sample volumes and allowed for the recovery of dead larvae from frozen samples. The sieve-PCR method enabled the identification of a single larva per 2 g of faecal sample and did not amplify DNA of a range of canine helminths, thus presenting a non-invasive tool for wildlife surveillance and for confirmative diagnosis in dogs.

The ADAMTS (a disintegrin and metalloproteinase domain with thrombospondin motifs) family includes 19 secreted proteinases in man. ADAMTS16 is a recently cloned gene expressed at high levels in fetal lung and kidney and adult brain and ovary. The ADAMTS-16 protein currently has no known function. ADAMTS16 is also expressed in human cartilage and synovium where its expression is increased in tissues from osteoarthritis patients compared to normal tissues. In this study, we ascertained that the full length ADAMTS16 mRNA was expressed in chondrocytes and cloned the appropriate cDNA. Stable over-expression of ADAMTS16 in chondrosarcoma cells led to a decrease in cell proliferation and migration, though not adhesion, as well as a decrease in the expression of matrix metalloproteinase-13 (MMP13). The transcription start point of the human ADAMTS16 gene was experimentally identified as 138 bp upstream of the translation start ATG and the basal promoter was mapped out to -1802 bp. Overexpression of Egr1 induced ADAMTS16 promoter constructs of -157/+138 or longer whilst Sp1 induced all ADAMTS16 promoter constructs. Transforming growth factor beta (TGFbeta) stimulated expression of endogenous ADAMTS16 gene expression in chondrocyte cell lines.

Trichinella surveillance in wildlife has relied on the detection of muscle larvae using digestion techniques. Serology has been proposed as more suitable for large-scale epidemiological studies in wildlife. In this study, 328 individual sera from wild red foxes and 16 sera from experimentally infected farmed foxes were serologically tested with both excretory/secretory antigen (E/S) and the synthetic beta-tyvelose glycan antigen, in indirect ELISA tests. The wild red foxes (Vulpes vulpes) had previously been examined for muscle larvae, using muscle digestion, whilst the experimentally infected farmed foxes were inoculated per os with either a low dose, 500 larvae, or a high dose, 10,000 of Trichinella nativa muscle larvae. Western blot (WB) was carried out on all seropositive samples using crude larval antigen. The present study found both beta-tyvelose and E/S antigen suited for the detection of antibodies to Trichinella spp., and T. nativa in particular, in foxes. Both ELISA antigens performed well, although, the E/S antigen was superior to the beta-tyvelose antigen, with sera that had been stored at -20 degrees C for more than 10 years. Neither antigen, however, detected all of the samples proven seropositive by WB: E/S detected 21 of the 27 wild red fox sera positive by WB; beta-tyvelose detected 22 positive sera; and in total 24 of the 27 positive WB sera were identified using both antigens. Serology alone, without WB or muscle digestion, led to a two- to threefold higher seroprevalence estimate, respectively. The use of E/S antigen in conjunction with the WB was the method of choice for the screening of wild red fox populations for Trichinella. Antibody persistence to T. nativa was short in the low dose group where antibody levels were not different from background by 32 wpi. In total, 7.3% (24/328) of the wild red fox population had antibodies to Trichinella on ELISA and WB. Antibodies were identified in foxes from a further two regions in Norway compared to the original muscle digestion results.

Echinococcus multilocularis, causing alveolar echinococcosis in humans, is a highly pathogenic emerging zoonotic disease in central Europe. The gold standard for the identification of this parasite in the main host, the red fox, namely identification of the adult parasite in the intestine at necropsy, is very laborious. Copro-enzyme-linked immunosorbent assay (ELISA) with confirmatory polymerase chain reaction (PCR) has been suggested as an acceptable alternative, but no commercial copro-ELISA tests are currently available and an in-house test is therefore required. Published methods for taeniid egg isolation and a multiplex PCR assay for simultaneous identification of E. multilocularis, E. granulosus and other cestodes were adapted to be carried out on pooled faecal samples from red foxes in Norway. None of the 483 fox faecal samples screened were PCR-positive for E. multilocularis, indicating an apparent prevalence of between 0% and 1.5%. The advantages and disadvantages of using the adapted method are discussed as well as the results pertaining to taeniid and non-taeniid cestodes as identified by multiplex PCR.

Introduction: The molecular mechanisms underlying cartilage destruction in osteoarthritis are poorly understood. Proteolysis is a key feature in the turnover and degradation of cartilage extracellular matrix where the focus of research has been on the metzincin family of metalloproteinases. However, there is strong evidence to indicate important roles for other catalytic classes of proteases, with both extracellular and intracellular activities. The aim of this study was to profile the expression of the majority of protease genes in all catalytic classes in normal human cartilage and that from patients with osteoarthritis (OA) using a quantitative method., Methods: Human cartilage was obtained from femoral heads at joint replacement for either osteoarthritis or following fracture to the neck of femur (NOF). Total RNA was purified, and expression of genes assayed using Taqman low-density array quantitative RT-PCR., Results: A total of 538 protease genes were profiled, of which 431 were expressed in cartilage. A total of 179 genes were differentially expressed in OA versus NOF cartilage: eight aspartic proteases, 44 cysteine proteases, 76 metalloproteases, 46 serine proteases and five threonine proteases. Wilcoxon ranking as well as the LogitBoost-NR machine learning approach were used to assign significance to each gene, with the most highly ranked genes broadly similar using each method., Conclusions: This study is the most complete quantitative analysis of protease gene expression in cartilage to date. The data help give direction to future research on the specific function(s) of individual proteases or protease families in cartilage and may help to refine anti-proteolytic strategies in OA.

The red fox (Vulpes vulpes) population, in Norway, was naïve to Sarcoptes scabiei prior to the late 1970s when this parasite was first recorded and a still ongoing epidemic started. During the course of this protracted epidemic some degree of host/parasite adaptation, with the occurrence of healthy antibody positive foxes, might be expected. In the present study the prevalence of sarcoptic mange and serologically identified S. scabiei exposure was investigated in 363 Norwegian red foxes, shot by hunters during two different study periods (1994-1995 and 2002-2005). The sarcoptic mange diagnosis was based upon the presence of clearly visible lesions in the skin of the cadaver with confirmatory demonstration of S. scabiei. The serodiagnosis was based on an indirect-ELISA. There was a significant decrease in prevalence of both mange cases and seropositive animals from the first to the second study period. Whilst the mange prevalence fell more than threefold, from 30.0% to 6.6%, the seroprevalence dropped less dramatically from 53.3% to 19.1%. The smaller decrease in seroprevalence compared to mange cases reflected a significantly higher ratio of seropositive-mange negative versus seropositive-mange positive foxes, during the second study period, 40:18, compared to the first, 14:18. These findings indicate that the red fox population is adapting to live with the parasite and that low-grade or sub-clinical infections, and even recoveries, occur amongst exposed foxes. Mange positive foxes had significantly poorer body condition than those without sarcoptic mange. No significant difference in body condition was seen between seropositive-mange negative versus seronegative-mange negative foxes. The ELISA sensitivity was found to be 95% and proved a useful tool for investigating the exposure to S. scabiei in wild foxes. This study is believed to be the first pointing to a long-term Sarcoptes/fox adaptation, combining long-term prevalence studies of clinical sarcoptic mange and serological evidence of exposure to the parasite in the general fox population.

Trichinella nativa is the most frequent Trichinella species in arctic wildlife and also the predominating species seen in Norwegian fauna. The adaptation of T. nativa to a cold climate is reflected by the well-documented freeze tolerance of its muscle larvae. The ability of the larvae to survive repeated freezing and thawing events has not however been elucidated and was investigated in the present study, using an Alaskan isolate and two isolates from coastal and inland Norway, respectively. Each T. nativa isolate was inoculated in foxes (Vulpes vulpes) and the muscle tissue obtained after 20 weeks was minced and divided into freezer bags. In the initial part of the study, the bags were exposed to either continuous freezing (-5 degrees C) for up to 7 weeks or freezing (-5 degrees C) for up to 7 weeks with seven overnight (+21 degrees C) thawing events. Once a week a bag was removed from each group, the meat was digested and muscle larvae isolated. In vitro assessment of larval viability was carried out based on larval motility (active vs non-active) and morphology, coiled (alive) or C-shaped (dead). Larval infectivity was subsequently bioassayed in mice, administering 500 larvae per mouse. The mice were euthanised 4 weeks post inoculation, the muscle digested and larvae per gram (lpg) and reproductive capacity index (RCI) were calculated. During the second part of the study, some of the minced fox muscle, exposed to the initial freeze protocol, was stored for a further 23 weeks at -18 degrees C prior to in vitro and in vivo assessment of larval viability and infectivity. The study demonstrated that Trichinella isolates originating from carnivores from higher northern latitudes expressed highest tolerance to freezing and that temperature fluctuations around freezing point, for up to 7 weeks, had little effect on larval infectivity. A negative effect of the initial repeated freeze-thaw events could be demonstrated once the larvae were exposed to longer periods of subsequent deep freezing. Furthermore, it was demonstrated that larval morphology and motility are not suitable for the assessment of infectivity of Trichinella larvae. It was concluded that bioassay in mice was the only suitable method currently available for assessing larval viability.

Purpose: Dupuytren's disease (DD) is a common fibrotic condition of the palmar fascia, leading to deposition of collagen-rich cords and finger contractions. The metzincin superfamily contains key enzymes in the turnover of collagen and other extracellular matrix macromolecules. A number of broad-spectrum matrix metalloproteinase inhibitors, used in cancer clinical trials, caused side effects of DD-like contractures. We tested the hypothesis that changes in the expression of specific metalloproteinases underlie or contribute to the fibrosis and contracture seen in DD., Methods: We collected tissue from patients with DD and used normal palmar fascia as a control. We profiled the expression of the entire matrix metalloproteinase (MMP), tissue inhibitor of metalloproteinases (TIMP), and a disintegrin and metalloproteinase domain with thrombospondin motif (ADAMTS) gene families in these tissues using real-time reverse-transcription polymerase chain reaction., Results: A number of metalloproteinases and inhibitors are regulated in DD. The expression of 3 key collagenases, MMP1, MMP13, and MMP14 is increased significantly in the DD nodule, as is the expression of the collagen biosynthetic enzyme ADAMTS14. The expression of MMP7, an enzyme with broad substrate specificity, is increased in the DD nodule and remains equally expressed in the DD cord. TIMP1 expression is increased significantly in DD nodule compared with normal palmar fascia., Conclusions: This study measured the expression of all MMP, ADAMTS, and TIMP genes in DD. Contraction and fibrosis may result from: (1) increased collagen biosynthesis mediated by increased ADAMTS-14; (2) an increased level of TIMP-1 blocking MMP-1- and MMP-13-mediated collagenolysis; and (3) contraction enabled by MMP-14-mediated pericellular collagenolysis (and potentially MMP-7), which may escape inhibition by TIMP-1. The complete expression profile will provide a knowledge-based approach to novel therapeutics targeting these genes.

The first report of Aelurostrongylus falciformis (Schlegel 1933) in Fennoscandian badgers is described. Routine parasitological examination of nine Norwegian badgers, at the National Veterinary Institute during 2004 and 2005, identified A. falciformis in the terminal airways of five of the animals. The first stage larvae (L1) closely resembled, in size and morphology, those of Angiostrongylus vasorum (Baillet 1866). The diagnosis for both A. falciformis and A. vasorum is frequently based on the identification of L1 in faeces or sputum. The potential for misclassification of an A. falciformis infection as A. vasorum, where larval identification is the only diagnostic method used, is discussed.

A survey of the parasitic fauna of the Norwegian red fox (Vulpes vulpes) population was carried out in 1994/1995 and 2002-2005. All foxes were killed during the licensed hunting season from October to April and, in total, 393 animals from all regions of the country were examined. The present study details the results of extra-intestinal nematode and Trichinella larvae examinations. All individuals were examined for Trichinella, using routine digestion methods. Parasitological examination of the internal organs of some of the foxes also identified a number of different extra-intestinal nematodes. The following prevalences were identified (number positive/number foxes examined): Trichinella larvae 19/393 (4.8%); Capillaria böhmi (C. böhmi) 88/174 (51%); Capillaria aerophila (C. aerophila) 160/181 (88%); Crenosoma vulpis (Cr. vulpis) 105/181 (58%) and Capillaria plica (C. plica) 81/154 (53%). No evidence of Angiostrongylus vasorum infection was found. The 19 different Trichinella isolates were species typed by PCR and sequence analysis; 18 isolates were identified as Trichinella nativa and one as Trichinella britovi. A wide geographical distribution of the parasites was seen. The following exceptions were recorded: C. böhmi, the prevalence of which was significantly lower in northern Norway (6%) compared to other regions (central Norway, eastern Norway and southern and western Norway; 52-57%). There was a significantly higher prevalence of Trichinella infection in eastern Norway (8.1%), when compared with the rest of the country (0.6%). Cr. vulpis prevalence was significantly higher in central Norway (83%) than in other regions (41-56%). There were no significant differences in age and sex distribution of the parasites with the exception of Cr. vulpis where juvenile foxes had a greater likelihood of infection. The data also indicated that adult foxes were more commonly infected with Trichinella larvae (5.8%) than juveniles (3.3%) (no statistical significance).

Cartilage destruction in osteoarthritis (OA) is thought to be mediated by two main enzyme families; the matrix metalloproteinases (MMPs) are responsible for cartilage collagen breakdown, whereas enzymes from the 'a disintegrin and metalloproteinase domain with thrombospondin motifs' (ADAMTS) family mediate cartilage aggrecan loss. Tissue inhibitors of metalloproteinases (TIMPs) regulate the activity of these enzymes. Although cartilage destruction in OA might be driven by the chondrocyte, low-grade synovitis is reported in patients with all grades of this disease. Our earlier work profiling these gene families in cartilage identified a number of genes that are regulated in OA, which are hence implicated in the disease process. Because the synovium might contribute to cartilage-matrix destruction in OA, we have extended the screening in the current study. We have profiled MMP, ADAMTS and TIMP genes in both cartilage and synovium from patients with either OA of the hip or a fracture to the neck of femur (NOF), giving a more complete picture of proteolysis in this disease. The four most significantly upregulated genes (P < 0.0001) in OA synovium compared to the fractured NOF are MMP28, ADAMTS16, ADAMTS17 and TIMP2. For MMP9, MMP10, MMP12, MMP17, MMP23, MMP28, ADAMTS4, and ADAMTS9, there is a significant correlation between expression levels in the synovium and cartilage, suggesting similar mechanisms of regulation. Additionally, we have shown that in cartilage the median level of steady-state mRNA for MMP13 is approximately 20-fold higher than MMP28 and approximately 1,500-fold higher than ADAMTS16, with expression of this latter gene approximately 150-fold higher in synovium than cartilage. This study is the most comprehensive analysis of the metzincin family of proteinases in the joint to date and has identified several proteinase genes not previously reported to be expressed or regulated in synovium.

1. The authors attempted to detect a possible effect of treatment with phenelzine on a physiological response to nicotine in the rat. 2. Positive findings in an animal model suggest the feasibility of more complicated experiments in animals and the possibility of studies involving human subjects. 3. Treatment of Sprague Dawley rats (n = 10) with phenelzine sulfate (15.0 mg/kg ip) every 48 hours for 14 days was associated with a 73.3% decrease in the hypothermic response to nicotine. 4. Treatment with phenelzine did not enhance the rate of elimination of nicotine. 5. The authors discuss a possible relationship between changes in nicotinic mechanisms and the therapeutic actions of drugs used to treat affective illness.

1. Chronic treatment with amitriptyline produces supersensitivity to the hypothermic effects of the muscarinic agonist oxotremorine. 2. Chronic treatment with amitriptyline also produces supersensitivity to the hypothermic effects of nicotine. 3. Oxotremorine and other naturally occurring muscarinic agonists are also nicotinic agonists. 4. Chronic treatment with amitriptyline produces time-dependent and reversible supersensitivity to the hypothermic effects of nicotine. 5. The authors assessed the possibility that the development of supersensitivity to oxotremorine following chronic treatment with amitriptyline is related to an effect of this antidepressant on a nicotinic mechanism. 6. A nicotinic receptor antagonist would blunt (though not necessarily eliminate) enhanced sensitivity to the thermic effects of oxotremorine if the nicotinic effects of the latter are significant. 7. The simultaneous administration of mecamylamine (a peripherally and centrally active nicotinic receptor antagonist) greatly augments (rather than blunts) the hypothermic response to oxotremorine. 8. The data suggest that the oxotremorine may activate a nicotinic mechanism counterbalancing its effect on a muscarinic mechanism. 9. This is consistent with previously published reports that the activation of nicotinic and muscarinic mechanisms can exert opposite effects.

Parasitic diseases are primarily diseases of poverty. At serious risk are individuals, communities and countries least able to afford the costs of treatment or prevention. In turn, economic development projects which aim to increase income levels may lead to negative results because of increased transmission of parasitic diseases often results. In attempting to analyse the economic consequences of parasitic diseases and the economics of their control, economists have usually relied on the tools of cost-benefit analysis, cost-effectiveness analysis and financial analysis. These efforts are briefly reviewed in the paper. The results have been subject to considerable criticism because of conceptual and methodological problems. For example, most studies have not taken into account the epidemiology and natural history of the disease in estimating the associated economic losses, thereby leading to inappropriate conclusions. The UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases through its Social and Economic Research Scientific Working Group (SER-SWG) is promoting research on the economics of parasitic diseases. In an attempt to improve the usefulness and reliability of such studies, conceptual and methodological approaches have been suggested which are reported on here. To improve the research design of these projects, a conceptual framework is proposed which consists of four components: (1) baseline social, cultural and economic conditions influencing disease transmission; (2) resources already invested in the health system and investments in other related sectors such as agriculture, housing, water supply and sanitation; (3) health consequences resulting from (1) and (2); and (4) social and economic consequences resulting from (3). A key concern in relation to the framework is to determine the most useful basis for linking results from studies of one component to another so as to analyse more systematically the impacts of disease on individuals and society. Such studies are inherently interdisciplinary and close collaboration of economists with medical scientists and disease control programme staff is needed to ensure completeness and reliability of input data and results. Results from these studies could be used to inform national decision-makers about the social and economic consequences of the parasitic diseases and their control and, thus, should strengthen support for increased investment to reduce the parasitic disease burden in developing countries.

Aneurysms of the extracranial carotid arteries are rare vascular lesions that produce a high incidence of unfavorable neurologic sequelae. In the past 24 years, 19 patients at UCLA Hospital were treated for angiographically demonstrable extracranial carotid arterial aneurysms. These cases were reviewed to determine the natural history of these lesions as influenced by various modes of therapy in order to develop a logical treatment plan for these difficult patients. Because of the varied location of these lesions, proper treatment requires a diversity of techniques. Gratifying results can be achieved when a carefully selected operative approach is undertaken.

Desipramine HCl, 10 mg/kg i.p. twice daily, produced subsensitivity to the hypothermic effects of nicotine, 1 mg/kg, after 1 and 2 weeks of treatment in male Sprague-Dawley rats, Phenelzine sulfate, fluoxetine HCl, and bright artificial light produced the same effect. The capacity of three chemically distinct classes of anti-depressants and bright artificial light (a treatment for seasonal depression) to produce this result suggests that effects on nicotinic mechanisms may be involved in the mechanism of action of these treatments.

Fluoxetine HCl, 10 mg/kg ip, twice daily produced subsensitivity to the hypothermic effects of nicotine (base), 1 mg/kg ip, after 1 (p less than 0.02) and 2 (p less than 0.002) weeks of treatment. Phenelzine sulfate, desipramine HCl and bright artificial light produced the same effect. The capacity of three chemically distinct classes of antidepressants and bright artificial light (a treatment for seasonal depression) to produce this result suggests that effects on nicotinic mechanisms may be involved in the mechanism of action of these treatments.

Two hundred fifteen patients with a history of either stroke, transient ischemic attack (TIA), or asymptomatic carotid bruit underwent noninvasive carotid artery testing using oculopneumoplethysmography. Of patients with hemodynamically significant stenosis, 51 (40.8%) underwent endarterectomy, and 74 (59.2%) were treated nonoperatively. The incidence of stroke in the nonoperated group was 12/74 (16.2%) compared with only 1/51 (1.9%) in the operated group. Similarly, recurrent TIA occurred in 29/74 (39.2%) of the nonoperated group vs 9/51 (17.6%) of the operated. In nonhemodynamically significant carotid stenosis, the risk of cerebrovascular death and stroke was exceedingly low: 2/90 (2.2%). Patients with hemodynamically significant stenosis treated nonoperatively have a greater risk of cerebrovascular death, stroke, and TIA than patients treated with carotid endarterectomy.

1. Two but not one week of treatment with amitriptyline (AMI) produces subsensitivity to the hypothermic effects of the alpha 2 antagonist yohimbine. 2. Subsensitivity persisted for the three weeks during which it was measured following the discontinuation of AMI. 3. Two weeks of twice daily injections of saline did not alter the thermic response to this agent. 4. The results support other data indicating tricyclic antidepressants (TCAs) subsensitize alpha 2 mediated thermic changes and are consistent with reports that TCAs down-regulate alpha 2 receptors.

The release of endogenous catecholamines (CA) from rat brain slices containing the nucleus tractus solitarius (NTS) was measured using a sensitive radioenzymatic assay. KCl (35 to 75 mM) induced a dose-related increase in norepinephrine (NE) release. Dopamine (DA) release was maximal with 50 mM KCl. An increase in epinephrine (E) release was only observed with 75 mM CKl. NE and E release was totally calcium-dependent whereas DA release was only partially calcium-dependent. Subsequent administrations of KCl released less CA. The calcium dependency of the KCl induced released of E, NE, and DA suggests a neurotransmitter function in the NTS for these CA. A difference in storage sites and/or mechanisms may be responsible for the observed differences in sensitivity to KCl and to extracellular calcium.

A prospective, randomized, double-blind clinical study was performed to determined the efficacy of short-term (24 hr) perioperative antibiotics in preventing septic complications after emergency appendectomy for nonperforated appendicitis. The patients were stratified into three clinical arms: Group I (placebo, n = 45), Group II (cefamandole, n = 46) and Group III (cefamandole plus carbenicillin, n = 45). The three groups of patients were similar in regard to age, sex, duration of operation and pathologic classification of the appendix. The overall incidence of infection in the study was 5.1%. The infection rates in Groups II (2.2%) and III (0%) were significantly lower than Group I (placebo) (13.3%), (p less than 0.05). No difference was observed between cefamandole alone and cefamandole plus carbenicillin. Average postoperative hospital days per patient for each group was: Group I - 3.8 days; Group II - 2.9 days; Group III - 3.1 days. Cost analysis of hospitalization including cost of prophylactic antibiotics revealed a $247.99 per patient saving for Group II versus Group I and $95.53 for Group III versus Group I. Systemic prophylactic antibiotics can successfully reduce septic complications after appendectomy for nonperforated appendicitis, and a single drug (cefamandole) directed at the facultative pathogens is as effective as double drug therapy, which includes specific anaerobic coverage.