Differential characterization of emerging skin diseases of rainbow trout--a standardized approach to capturing disease characteristics and development of case definitions.

Farmed and wild salmonids are affected by a variety of skin conditions, some of which have significant economic and welfare implications. In many cases, the causes are not well understood, and one example is cold water strawberry disease of rainbow trout, also called red mark syndrome, which has been recorded in the UK since 2003. To date, there are no internationally agreed methods for describing these conditions, which has caused confusion for farmers and health professionals, who are often unclear as to whether they are dealing with a new or a previously described condition. This has resulted, inevitably, in delays to both accurate diagnosis and effective treatment regimes. Here, we provide a standardized methodology for the description of skin conditions of rainbow trout of uncertain aetiology. We demonstrate how the approach can be used to develop case definitions, using coldwater strawberry disease as an example.

Content variability of active drug substance in compounded oral 3,4-diaminopyridine products.

WHAT IS KNOWN AND OBJECTIVE: 3,4-diaminopyridine (3,4-DAP; amifampridine) is used for symptomatic treatment of Lambert-Eaton myasthenic syndrome. Until recently, it was only available as a compounded product, which raises safety concerns because of possible high variability in active drug substance content. The objective of this study was to evaluate the variability in dosage form weight, active content variability and impurity of compounded oral 3,4-DAP drug products. METHODS: Ten samples each of 9 oral 3,4-DAP compounded products were weighed, extracted with water and the 3,4-DAP content determined by ultra high-performance liquid chromatography. RESULTS AND DISCUSSION: Variability in dosage form weight ranged from 0·81% relative standard deviation (RSD) to 4·82% RSD. In the 90 samples tested, 3,4-DAP content ranged from 22·2% to 125·2% of declared label content. All 10 samples of one compounded product had active drug substance content well below the declared label content (35·0%, 51·7% RSD). No compounded product achieved the Good Manufacturing Practice (GMP) standard of 95-105% range limit of declared label content; one achieved 90-110%, and four others achieved 80-120% of declared content for all 10 samples. There was no evidence of a significant presence of degradation products or related substances in any compounded product. WHAT IS NEW AND CONCLUSION: Compounded 3,4-DAP products are subject to considerable variability in active drug substance content. This variability seems to be principally because of heterogeneous formulated material rather than variation in dosage form weight.

The potential for targeted surveillance of live fish movements in Scotland.

The network structure of the movements of live fish in the Scottish aquaculture industry has recently been demonstrated for 2003. In this paper, we enlarge this analysis to a longer 3-year period from 2002 to 2004, the new data allowing complete coverage of at least one production cycle. The resulting network contains slightly more sites than that for a single year and is denser with more arcs (directed site-to-site connections) present, but otherwise features recognizable in the 1-year network are still recognizable in the 3-year network. Arc-removal algorithms (a proxy for targeted surveillance) were identified that could successfully reduce the portion of the network reachable from a node (a proxy for potential epidemic size) by approximately one-third by removing as few as four arcs. This results from the high centrality of particular nodes and arcs. A strong community structure was identified in the network, corresponding with species farmed, but only weakly geographical, with a high proportion of arcs occurring between management areas and catchments.

In vitro human skin absorption of linalool: Effects of vehicle composition, evaporation and occlusion on permeation and distribution.

In vitro human skin permeation and distribution of the fragrance material linalool (3,7-dimethyl-1,6-octadien-3-ol, CAS No. 78-70-6) following application in a range of single and mixed vehicles was determined, under unoccluded and occluded conditions, using human epidermal membranes. Vehicles were (70/30 v/v) ethanol[EtOH]/water, dipropyleneglycol [DPG], diethyl phthalate [DEP], (25/75 v/v) EtOH/DEP, (25/75 v/v) EtOH/DPG and petrolatum. Worst case absorbed dose values (% applied dose) for linalool under unoccluded conditions varied from 1.84% (DPG) to 4.08% (EtOH/water) and under occluded conditions from 5.9% (DEP) to 14.7% (EtOH/water). Occlusion always increased absorption but the magnitude of the effect varied with the vehicle from 2 to 6-fold. This study demonstrated that in vitro human skin permeation of linalool varied quite widely between test vehicles and that the magnitude of the effect of occlusion was also vehicle dependent. This was particularly significant in view of the reported variations in biological responses using different vehicles (Lalko et al., 2004; Politano et al., 2006).

Seasonality and heterogeneity of live fish movements in Scottish fish farms.

Movement of live animals is a key contributor to disease spread. Farmed Atlantic salmon Salmo salar, rainbow trout Onchorynchus mykiss and brown/sea trout Salmo trutta are initially raised in freshwater (FW) farms; all the salmon and some of the trout are subsequently moved to seawater (SW) farms. Frequently, fish are moved between farms during their FW stage and sometimes during their SW stage. Seasonality and differences in contact patterns across production phases have been shown to influence the course of an epidemic in livestock; however, these parameters have not been included in previous network models studying disease transmission in salmonids. In Scotland, farmers are required to register fish movements onto and off their farms; these records were used in the present study to investigate seasonality and heterogeneity of movements for each production phase separately for farmed salmon, rainbow trout and brown/sea trout. Salmon FW-FW and FW-SW movements showed a higher degree of heterogeneity in number of contacts and different seasonal patterns compared with SW-SW movements. FW-FW movements peaked from May to July and FW-SW movements peaked from March to April and from October to November. Salmon SW-SW movements occurred more consistently over the year and showed fewer connections and number of repeated connections between farms. Therefore, the salmon SW-SW network might be treated as homogeneous regarding the number of connections between farms and without seasonality. However, seasonality and production phase should be included in simulation models concerning FW-FW and FW-SW movements specifically. The number of rainbow trout FW-FW and brown/sea trout FW-FW movements were different from random. However, movements from other production phases were too low to discern a seasonal pattern or differences in contact pattern.

A panoramic code for sound location by cortical neurons.

By conventional spike count measures, auditory neurons in the cat's anterior ectosylvian sulcus cortical area are broadly tuned for the location of a sound source. Nevertheless, an artificial neural network was trained to classify the temporal spike patterns of single neurons according to sound location. The spike patterns of 73 percent of single neurons coded sound location with more than twice the chance level of accuracy, and spike patterns consistently carried more information than spike counts alone. In contrast to neurons that are sharply tuned for location, these neurons appear to encode sound locations throughout 360 degrees of azimuth.

The contact structure of Great Britain's salmon and trout aquaculture industry.

We analyse the network structure of the British salmonid aquaculture industry from the perspective of infectious disease control. We combine for the first time live fish transport (or movement) data covering England and Wales with data covering Scotland and include network layers representing potential transmission by rivers, sea water and local transmission via human or animal vectors in the immediate vicinity of each farm or fishery site. We find that 7.2% of all live fish transports cross the England-Scotland border and network analysis shows that 87% of English and Welsh nodes and 72% of Scottish nodes are reachable from cross-border connections via live fish transports alone. Consequently, from a disease-control perspective, the contact structures of England and Wales and of Scotland should not be considered in isolation. We also show that large epidemics require the live fish movement network and so control strategies targeting movements can be very effective. While there is relatively low risk of widespread epidemics on the live fish transport network alone, the potential risk is substantially amplified by the combined interaction of multiple network layers.

Identifying genotype specific elevated-risk areas and associated herd risk factors for bovine tuberculosis spread in British cattle.

Bovine tuberculosis (bTB) is a chronic zoonosis with major health and economic impact on the cattle industry. Despite extensive control measures in cattle and culling trials in wildlife, the reasons behind the expansion of areas with high incidence of bTB breakdowns in Great Britain remain unexplained. By balancing the importance of cattle movements and local transmission on the observed pattern of cattle outbreaks, we identify areas at elevated risk of infection from specific Mycobacterium bovis genotypes. We show that elevated-risk areas (ERAs) were historically more extensive than previously understood, and that cattle movements alone are insufficient for ERA spread, suggesting the involvement of other factors. For all genotypes, we find that, while the absolute risk of infection is higher in ERAs compared to areas with intermittent risk, the statistically significant risk factors are remarkably similar in both, suggesting that these risk factors can be used to identify incipient ERAs before this is indicated by elevated incidence alone. Our findings identify research priorities for understanding bTB dynamics, improving surveillance and guiding management to prevent further ERA expansion.

In-vitro human skin penetration of the fragrance material geranyl nitrile.

In-vitro human skin permeation and distribution of geranyl nitrile (GN) was determined using epidermal membranes following application (5 microl/cm(2)) in 70% ethanol, under non-occlusive conditions, at maximum in-use concentration (1%). Permeation was measured (12 time-points over 24 h) using 6% (w/v) Oleth-20 in pH 7.4 phosphate buffered saline as receptor. Permeation of reference benzoic acid was assessed using the same skin donors. Overall recovery of GN at 24 h was low (14.1+/-0.4%) due to evaporation. Evaporative loss of GN from polytetrafluoroethylene (PTFE) sheet, under the same conditions was rapid (93% over 24h) although this overestimated loss during permeation where evaporation competed with uptake. At 24 h, 1.89+/-0.15 microg/cm(2) GN, (3.74+/-0.30% of applied dose) (mean+/-standard error, SE, n=12), had permeated. Following rapid initial permeation, the absorption plateaued due to depletion. Levels of GN in the epidermis (plus any remaining stratum corneum after tape stripping), filter paper membrane support and receptor fluid were combined (as per SCCNFP guidelines) to produce a total absorbed dose value of 4.72+/-0.32%. Systemic exposure resulting from the use of GN as a fragrance ingredient, under unoccluded conditions, would be low based on the currently reported use levels.

Data quality of the Cattle Tracing System in Great Britain.

The Cattle Tracing System (cts) of Great Britain was examined to investigate the quality of its data in terms of known errors and omissions, and their distribution. The proportions of erroneous or missing data have decreased steadily over time, with a marked improvement in the quality of the data since 2001, when recording became mandatory. There is little variation between regions in the quality of the data, but there are potentially important variations between the types of agricultural premises that cattle move to and from, and in other factors correlated with the type of premises, such as the age of the animals and the number of animal movements associated with the premises.

Heterogeneous responses of temperate-zone amphibian populations to climate change complicates conservation planning.

The pervasive and unabated nature of global amphibian declines suggests common demographic responses to a given driver, and quantification of major drivers and responses could inform broad-scale conservation actions. We explored the influence of climate on demographic parameters (i.e., changes in the probabilities of survival and recruitment) using 31 datasets from temperate zone amphibian populations (North America and Europe) with more than a decade of observations each. There was evidence for an influence of climate on population demographic rates, but the direction and magnitude of responses to climate drivers was highly variable among taxa and among populations within taxa. These results reveal that climate drivers interact with variation in life-history traits and population-specific attributes resulting in a diversity of responses. This heterogeneity complicates the identification of conservation 'rules of thumb' for these taxa, and supports the notion of local focus as the most effective approach to overcome global-scale conservation challenges.

Modelling the initial spread of foot-and-mouth disease through animal movements.

Livestock movements in Great Britain (GB) are well recorded and are a unique record of the network of connections among livestock-holding locations. These connections can be critical for disease spread, as in the 2001 epidemic of foot-and-mouth disease (FMD) in the UK. Here, the movement data are used to construct an individual-farm-based model of the initial spread of FMD in GB and determine the susceptibility of the GB livestock industry to future outbreaks under the current legislative requirements. Transmission through movements is modelled, with additional local spread unrelated to the known movements. Simulations show that movements can result in a large nationwide epidemic, but only if cattle are heavily involved, or the epidemic occurs in late summer or early autumn. Inclusion of random local spread can considerably increase epidemic size, but has only a small impact on the spatial extent of the disease. There is a geographical bias in the epidemic size reached, with larger epidemics originating in Scotland and the north of England than elsewhere.

Demographic structure and pathogen dynamics on the network of livestock movements in Great Britain.

Using a novel interpretation of dynamic networks, we analyse the network of livestock movements in Great Britain in order to determine the risk of a large epidemic of foot-and-mouth disease (FMD). This network is exceptionally well characterized, as there are legal requirements that the date, source, destination and number of animals be recorded and held on central databases. We identify a percolation threshold in the structure of the livestock network, indicating that, while there is little possibility of a national epidemic of FMD in winter when the catastrophic 2001 epidemic began, there remains a risk in late summer or early autumn. These predictions are corroborated by a non-parametric simulation in which the movements of livestock in 2003 and 2004 are replayed as they occurred. Despite the risk, we show that the network displays small-world properties which can be exploited to target surveillance and control and drastically reduce this risk.

Effect of early full weight-bearing after joint injury on inflammation and cartilage degradation.

BACKGROUND: Early full weight-bearing after an acute osteochondral injury avoids problems associated with immobility but may also be harmful by amplifying the inflammatory response. To investigate these effects, we developed an in vivo model of subchondral trauma. METHODS: After an impact injury to the femoral condyle, fourteen dogs were randomized to immediate full weight-bearing or to four weeks of minimal weight-bearing before full weight-bearing. Synovial fluid was sampled by aspiration at one, two, four, eight, twelve, sixteen, twenty, and twenty-four weeks. Neutrophils, monocytes, and lymphocytes were enumerated, and the concentrations of tumor necrosis factor-alpha, interleukin-10, nitric oxide, matrix metalloproteinases, and glycosaminoglycans were measured. RESULTS: Compared with the findings for uninjured joints, the synovial fluid from the impacted joints of full-weight-bearing dogs had significantly higher peak concentrations of neutrophils (p = 0.0006 at one week), mononuclear leukocytes (p = 0.001 at four weeks), tumor necrosis factor-alpha (p = 0.001 at one week), nitric oxide (p = 0.001 at one week), matrix metalloproteinases (p = 0.008 at one week), and glycosaminoglycans (p = 0.002 at four weeks and p = 0.001 at six months). The size of the bone bruise correlated with the peak concentrations of tumor necrosis factor-alpha (r2= 0.89, p = 0.007; Spearman rank test), matrix metalloproteinases (r2= 0.96, p = 0.0004), and glycosaminoglycans (r2= 0.96, p = 0.0004). However, restriction to minimal weight-bearing for four weeks after the injury led to a significant reduction in the synovial fluid concentrations of neutrophils (p = 0.007 at one week and p = 0.01 at two weeks), tumor necrosis factor-alpha (p = 0.0006 to 0.02 during the first four weeks), nitric oxide (p = 0.001 to 0.04 during the first four weeks), and matrix metalloproteinases (p = 0.007 to 0.01 from the second week to the eighth week). In contrast, interleukin-10 concentrations were significantly higher (p = 0.002 at one week) and glycosaminoglycan levels remained at normal levels in animals that were restricted from immediate full weight-bearing after the injury. CONCLUSIONS: The magnitude of the inflammatory response is proportional to the size of the bone bruise. Restriction to minimal weight-bearing for four weeks reduces the magnitude of the inflammatory response and the cartilage degradation following articular cartilage impact injury. CLINICAL RELEVANCE: Strategies to minimize mechanical stress during the early postinjury period may help to preserve cartilage integrity and forestall the development of osteoarthritis.

Second neoplasms after Wilms' tumor in childhood.

The frequency of second neoplasms was examined among 487 Wilms' tumor patients treated at the Dana-Farber Cancer Institute and Children's Hospital of Boston, 1927-81. Thirty study patients (6%) developed second primary tumors: 11 cancers, 16 benign tumors, and 3 borderline neoplasms. Cumulative probability of a second cancer was 18% (standard error, 6%) in 34 years after diagnosis of Wilms' tumor. The subgroup of 412 patients who had received radiotherapy for Wilms' tumor developed all 11 second cancers, which included 1 skin carcinoma, 1 acute leukemia, and 9 solid internal cancers (expected, 0.7 cancers other than skin carcinoma; P less than .001). After exclusion of the secondary leukemia, all but 1 second cancer arose within the radiotherapy field. Concurrent therapy with dactinomycin did not reduce the risk of a radiation-associated cancer. Second cancer was the cause of death in 7 patients.

Renal failure in the Denys-Drash and Wilms' tumor-aniridia syndromes.

Nearly 6000 patients enrolled in four clinical trials of the National Wilms' Tumor Study Group during 1969-1995 were followed until death or for a median of 11.0 years of survival for the onset of renal failure (RF). Thirteen of 22 patients with Denys-Drash syndrome and 10 of 46 patients with the Wilms' tumor aniridia syndrome developed RF. The cumulative risks of RF at 20 years from Wilms' tumor diagnosis were 62% and 38%, respectively. Only 21 cases of RF were observed among 5358 patients with unilateral disease who did not have characteristic congenital genitourinary anomalies, and their risk was <1%. Although other explanations cannot be completely excluded, the high rate of RF in patients with the aniridia syndrome challenges the view that nephropathy is associated uniquely with missense mutations in the WT1 gene. It suggests the possibility of a further gradation in the spectrum of phenotypes associated with different WT1 mutations. Patients with Wilms' tumor and aniridia or genitourinary abnormalities should be followed closely throughout life for signs of nephropathy or RF.

Chemopreventive activity of tamoxifen, N-(4-hydroxyphenyl)retinamide, and the vitamin D analogue Ro24-5531 for androgen-promoted carcinomas of the rat seminal vesicle and prostate.

We evaluated the ability of dietary N-(4-hydroxyphenyl)retinamide; 1 alpha,25-dihydroxy-16-ene-23-yne-26,27-hexafluorocholecalcifero l (Ro24-5531); and tamoxifen to inhibit the development of androgen-promoted carcinomas of the accessory sex organs of male Lobund-Wistar rats. Invasive carcinomas of the seminal vesicle (SV) and anterior prostate (AP) were induced in Lobund-Wistar rats with three different combinations of initiator [N-nitroso-N-methylurea (NMU)] and promoter [testosterone propionate (TP)]: (a) high-dose NMU (30 mg/kg) + high-dose TP (20 mg via implant every 2 months); (b) high-dose NMU + low-dose TP (10 mg implanted every 2 months); or (c) low-dose NMU (15 mg/kg) + low-dose TP. During the period of TP administration, rats were fed a diet supplemented with either N-(4-hydroxyphenyl)retinamide (1 or 2 mmol/kg diet), Ro24-5531 (1.25 or 2.5 nmol/kg diet), tamoxifen (0.5 or 5 mg/kg diet), or vehicle alone. After sacrifice at 8.5 or 11 months, the prostate-seminal vesicle complex from each rat was processed in toto and histologically staged as to the extent of tumor involvement. In animals given low-dose TP, all three agents were significantly effective at reducing the incidence of invasive carcinomas of the SV and, to a lesser degree, the AP. Of the three agents, tamoxifen given in high dose (5 mg/kg) had the strongest activity, reducing the occurrence of invasive SV carcinomas from 72-83% in controls to 6% (P = 0.0001) and the occurrence of invasive AP carcinomas from 50-72% to 18-22% (P < 0.05).

In situ detection of alkaline nuclease activity in cells infected with herpes simplex virus type 1 (HSV-1).

An in situ assay for detection of alkaline nuclease activities has been adapted to the herpes simplex virus type 1 (HSV-1) system. Six major nuclease activities which migrate with molecular weights of 90,000, 85,000, 80,000, 76,000, 71,000 and 65,000, and six minor species of molecular weights 87,000, 81,000, 57,000, 18,500, 17,500 and 16,500 were detected in lysates of HSV-1 infected cells following SDS-polyacrylamide gel electrophoresis and enzyme activation in situ. An ELISA assay and an immunoprecipitation study indicated that the six major HSV-induced nuclease species are virus-specific. Moreover, a reconstruction experiment in which 14C-labelled protein markers were incubated with mock- and HSV-infected cell lysates demonstrates that the nuclease fractions detected in situ were not due to endogenous proteolytic activity. The 80,000, 76,000, 71,000 and 65,000 species were first detected at 4 h post-infection, whereas all others were detectable by 6 h post-infection. The activities of the major cellular nucleases of molecular weights 50,000. 48,000 and 45,000 decreased as a function of time post-infection. The level of expression of each of the virus-induced species was dependent upon the multiplicity of infection, and all virus-induced activities exhibited biochemical properties characteristic of purified HSV-1 alkaline nuclease, including activation and inhibition by specifications. The 76,000 HSV-induced alkaline nuclease species was also demonstrated to possess endonucleolytic activity.

Treatment outcomes in patients less than 2 years of age with small, stage I, favorable-histology Wilms' tumors: a report from the National Wilms' Tumor Study.

PURPOSE: Retrospective analyses were performed to determine the effect of tumor weight and therapy modifications on outcome in patients less than 2 years of age with stage I favorable-histology Wilms' tumors. PATIENTS AND METHODS: The 4-year relapse-free and overall survival percentages for patients randomized to different treatment regimens in National Wilms' Tumor Studies (NWTS)-1, -2, and -3 were calculated and compared. RESULTS: The 4-year relapse-free survival percentages of patients whose specimen weight was less than 550 g were found to be 89.1% on NWTS-1, 96.0% on NWTS-2, and 93.2% on NWTS-3. There was no evidence that the relapse-free survival of these patients had improved over time (P value for trend = .99). The 4-year relapse-free survival percentage for similar age and stage patients whose specimen weight was 550 g or greater was significantly poorer than that of patients with smaller tumors (P = .02). CONCLUSION: Changes in the NWTS treatment regimens over a period of more than 20 years have not improved on the excellent prognosis of patients who are less than 2 years of age at diagnosis and who have a stage I, favorable-histology Wilms' tumor with specimen weight less than 550 g. These data could be used as the basis for a future trial in which a subgroup of such patients is treated with nephrectomy only.

Prognostic factors for children with recurrent Wilms' tumor: results from the Second and Third National Wilms' Tumor Study.

The characteristics of 367 stage I-IV National Wilms' Tumor Study (NWTS) children who relapsed after initial treatment for unilateral disease in the second and third NWTS trials (NWTS-2 and -3) were analyzed to identify features predictive of survival. Although modifications in initial therapy resulted in a lower rate of first relapse in these two studies compared with NWTS-1, all previously identified prognostic factors after relapse remained statistically significant predictors of survival. Tumor histology, length of initial remission, initial therapy with two v three drugs, and site of relapse each were independently predictive of postrelapse survival. The 3-year postrelapse survival for all 367 patients was 30% +/- 3%; however, subgroups classified by these prognostic factors were identified that had 3-year postrelapse survival rates of greater than 40%. These subgroups included patients who had tumors of favorable histology (FH) that recurred (1) only in the lungs, (2) in the abdomen when radiotherapy (RT) was not initially given, or (3) that were originally stage I, (4) that were originally treated with only two drugs, or (5) that recurred 12 months or more after diagnosis. These results were achieved with the use of standard treatments, ie, surgery, RT, and chemotherapy using dactinomycin (AMD), vincristine (VCR), and Adriamycin [( ADR] doxorubicin; Adria Laboratories, Columbus, OH). It is suggested that patients in these groups might be managed with aggressive use of conventional therapies until newer chemotherapeutic agents and drug combinations are shown to be more effective. Patients with adverse prognostic features at relapse have a poor survival expectancy with standard measures. Salvage attempts for these patients are better based on experimental approaches.