Gene expression profiles of some immune relevant genes from skin of susceptible and responding Atlantic salmon (Salmo salar L.) infected with Gyrodactylus salaris (Monogenea) revealed by suppressive subtractive hybridisation.

Suppressive subtractive hybridisation was used to examine the genetic basis of susceptibility and resistance of the Atlantic salmon (Salmo salar) to Gyrodactylus salaris infection. Selected immune relevant genes are listed and two genes, for myeloid leukemia differentiation protein (Mcl-1) and opioid growth factor receptor (OGFr), obtained from the susceptible salmon library were characterised. Both sequences showed high amino acid identity and similarity with human and mouse isoforms, and their possible involvement in the response of salmon to G. salaris is discussed. Quantitative reverse transcriptase-PCR was performed for both genes. Upregulation of Mcl-1 in B1 backcross salmon of the susceptible phenotypic category compared with resistant salmon was demonstrated. The possible relationship of the salmon Mcl-1 and cytokines (interleukin 1beta) in the G. salaris-induced host response is discussed. Potential involvement of OGFr in the depletion of mucous cells during prolonged and heavy G. salaris infection, via suppression of DNA synthesis and profound decrease in basal cell proliferation, is proposed. However, only two of six susceptible fish showed high upregulation of OGFr, which might indicate that its expression is localised to sites of wounds resulting from a heavy burden of G. salaris.

Development and application of real-time PCR for specific detection of Lepeophtheirus salmonis and Caligus elongatus larvae in Scottish plankton samples.

Lepeophtheirus salmonis and Caligus elongatus are important parasites of wild and cultured salmonids in the Northern Hemisphere. These species, generically referred to as sea lice, are estimated to cost the Scottish aquaculture industry in excess of pound 25 million per annum. There is great interest in countries such as Ireland, Scotland, Norway and Canada to sample sea lice larvae in their natural environment in order to understand lice larvae distribution and improve parasite control. Microscopy is currently relied on for use in the routine identification of sea lice larvae in plankton samples. This method is, however, limited by its time-consuming nature and requirement for highly skilled personnel. The development of alternative methods for the detection of sea lice larvae which might be used to complement and support microscopic examinations of environmental samples is thus desirable. In this study, a genetic method utilising a real-time PCR Taqman-MGB probe-based assay targeting the mitochondrial cytochrome oxidase I (mtCOI) gene was developed, which allowed species-specific detection of L. salmonis and C. elongatus larvae from unsorted natural and spiked plankton samples. Real-time PCR is a rapid, sensitive, highly specific and potentially quantitative technique. This study demonstrated its suitability for the routine identification of L. salmonis and C. elongatus in mixed plankton samples. The real-time PCR assay developed has considerable potential for use in complementing, supporting and reducing reliance on time-consuming conventional microscopic examination for the specific identification of sea lice larvae in plankton samples.

Rhabdomyolysis in the military: recognizing late-onset very long-chain acyl Co-A dehydrogenase deficiency.

Very long-chain acyl Co-A dehydrogenase deficiency, an inborn error of lipid metabolism, is commonly thought of as a disease of infancy or early childhood. However, several cases of late-onset very long-chain acyl Co-A dehydrogenase have been reported. This report of two military men who survived basic training before their disease presentation broadens the spectrum of late-onset disease, presents two previously unreported mutations, and demonstrates the fine line between athletic, active lifestyle and severe disease presentation.

Identification and characterisation of the genomic segment 7 of the infectious salmon anaemia virus genome.

The isolation and characterisation of a gene encoding the putative matrix proteins of infectious salmon anaemia virus (ISAV) is reported. Following identification of an ISAV-specific sequence from a cDNA library, RACE-PCR was used to identify a mRNA transcript of approximately 1.2 kb containing the ISAV consensus sequence GCTAAGA at the 5' end. Although the cDNA transcript and its putative protein product did not possess high homology with other orthomyxoviral sequences, similarity to a paramyxovirus fusion glycoprotein and viral cell surface proteins was identified. The size of this transcript suggested that it was derived from segment 7 of the ISAV genome and encoded the matrix proteins. Like syntenic segments of other orthomyxoviruses, this segment was shown to encode at least two matrix proteins, M1 and M2. The existence of M1 and M2 ISAV mRNA was demonstrated by RT-PCR and sequencing, with the M1 transcript being more abundant than the M2 in infected cell cultures, as is found in other orthomyxoviruses. Nucleotide sequence comparison of segment 7 of the ISAV genome from isolates of different geographic origin indicated it to be the one of the most variable of the ISAV genes characterised to date.

The impact of HIV disease on an Irish prison population.

Between January 1987 and January 1991, 168 known HIV-infected prisoners have been incarcerated in Dublin's Mountjoy prison. This figure constitutes 16.6% of the total HIV-infected population in the Republic of Ireland over the same period. One hundred and forty-one (84%) of these prisoners have attended the Department of Genitourinary Medicine, St James's Hospital, Dublin. This group displayed considerable morbidity from HIV-related disease. Respiratory tract infection was the most frequent complication seen. Much additional morbidity was directly attributable to intravenous drug use. A survey of a representative group of inmates revealed that 64.7% were diagnosed HIV-positive in prison. The mean length of time spent incarcerated since the diagnosis of HIV infection was 38.9 months. Twenty-nine of 34 individuals who answered a questionnaire were imprisoned for drug-related crimes and 32 of 34 prisoners admitted to parenteral drug use within the prison. As the HIV epidemic unfolds in Dublin, increasing numbers of prisoners with symptomatic HIV disease will spend time incarcerated in Mountjoy prison. This will pose a considerable burden on prison and hospital medical services alike.

Spatial and temporal variations in parasite prevalence and infracommunity structure in herring (Clupea harengus L.) caught to the west of the British Isles and in the North and Baltic Seas: implications for fisheries science.

Herring Clupea harengus L. viscera were examined for endoparasitic infections as part of a multidisciplinary stock identification project (WESTHER, EU Contract no. Q5RS-2002-01 056) which applied a range of stock discrimination techniques to the same individual fishes to obtain comparable results for multivariate analysis. Spawning and non-spawning adults, and juvenile herring were caught, over 3 years, by commercial and research vessels from numerous locations to the west of the UK and Ireland, along with control samples of spawning fish from the eastern Baltic Sea, and juveniles from sites in the eastern and western North Sea, and the north of Norway. The metacercariae of two renicolid digeneans (Cercaria pythionike and Cercaria doricha), one larval nematode (Anisakis simplex s.s.) and one larval cestode (Lacistorhynchus tenuis) were selected as tag species. Results were compared with those from herring collected between 1973 and 1982, which suggested remarkable stability in the parasite fauna of herring in the study area. These species were used to compare the parasite infracommunities of spawning herring. A significant variation in infracommunity structure was observed between different spawning grounds. These results suggest that the parasite fauna of herring are spatially variable but remain temporally stable in both the short and long term. Significant differences in prevalence and abundance of infections and comparisons of parasite infracommunity enabled the separation of putative herring stocks west of the British Isles. Distinctive patterns of parasite infection in two different spawning groups off the north coast of Scotland suggest that this area is occupied by two spawning populations, one recruiting from the west of Scotland, the other from outside this area, and most likely from the eastern North Sea. The distribution patterns of L. tenuis, C. doricha and C. pythionike suggest the potential for fish that spawn in three distinct International Council for the Exploration of the Seas (ICES) management units to be present in mixed aggregations found over the Malin Shelf, with significant implications for management in this area.

Isolation of a novel fish thymidylate kinase gene, upregulated in Atlantic salmon (Salmo salar L.) following infection with the monogenean parasite Gyrodactylus salaris.

Analysis of differential gene expression in salmon (Salmo salar) blood following infection with the monogenean parasite Gyrodactylus salaris, resulted in the isolation of a thymidylate kinase gene not previously described from fish and which showed similarity to an LPS-inducible thymidylate kinase gene isolated from mouse macrophages. This salmon TYKi-like gene may play a role in an innate generalised response to pathogen infection as it was upregulated in salmon following infection with the parasite, and also in response to injection with the immunostimulants LPS and Poly I:C, used to emulate bacterial and viral infections, respectively. The possible role of this gene in the biosynthesis of mitochondrial DNA in activated macrophages, in response to G. salaris infection is discussed.

Effect of alterations of dietary sodium on the severity of asthma in men.

BACKGROUND: There is some evidence of a positive association between increased dietary salt consumption and both increased bronchial reactivity and mortality from asthma in men. This study assesses the effects of alterations in dietary salt consumption on the clinical severity of asthma in adult male asthmatic patients. METHODS: A randomised, double blind, placebo controlled, crossover design was employed. Twenty seven mild to moderate asthmatic patients were established on a low sodium diet (80 mmol/day) at the end of a 4-5 day run in period and then randomised to receive 200 mmol/day slow sodium or matching placebo for five weeks, crossing over to the alternative regime for a further five weeks. Patients used diary cards to record twice daily peak expiratory flow rates, daily symptom scores, and bronchodilator consumption. Spirometry and degree of bronchial responsiveness (methacholine challenge test) were measured at screening and at the end of each treatment period. Twenty four hour urinary sodium excretion was measured at screening and in duplicate for each treatment period. RESULTS: Twenty two patients completed the study. For these patients the mean (95% confidence interval (CI)) difference in 24 hour sodium excretion between treatments was 204 (175 to 235) mmol. Compared with placebo, sodium supplementation resulted in deleterious alterations of all measured parameters. Bronchial reactivity rose on slow sodium with a 0.73 (0.2 to 1.3) doubling dose methacholine difference compared with placebo. Estimated median (95% CI) difference in bronchodilator consumption was 1.3 (0.4 to 2.1) puffs per day, the estimated median difference in symptom score was 0.6 (0.2 to 0.9), and mean forced expiratory volume in one second fell by 0.21 (0.05 to 0.37) 1. The peak expiratory flow rate rose on placebo and fell on slow sodium. Median differences between treatments were 5.6% (2.2% to 9.8%) for morning and 7.8% (3.9% to 12.9%) for evening peak expiratory flow rate. CONCLUSIONS: Our results suggest that large increases in dietary sodium result in physiological deterioration and increased morbidity in male asthmatic patients.

A study of acute asthma in the accident and emergency department.

We have conducted a survey of all the asthmatic patients seen at the Leicester Royal Infirmary Accident and Emergency Department during a 1-year period. A significantly greater number of patients were seen in the 8-hour period 1600-2359 hours and a greater number of patients were seen on Sundays and bank holidays. Wheeze was recorded in only 43% of the patients and there was no record of any nocturnal asthmatic symptoms. Peak expiratory flow rate was only recorded in 26% of patients before treatment with nebulized salbutamol (the commonest treatment). Pulse rate, respiratory rate, time of day or month did not determine whether or not a patient was admitted. We conclude that the education of junior doctors in the management of asthma is still deficient.

The effect of lifestyle on wheeze, atopy, and bronchial hyperreactivity in Asian and white children.

There is increasing evidence linking Western culture to increasing prevalence of asthma. We have measured the prevalence of symptoms of asthma, bronchial hyperreactivity, and atopy in a sample of white and Asian inner city schoolchildren aged 8-11 yr in the U.K. and assessed the influence of certain aspects of western culture on these variables. Bronchial reactivity was measured by methacholine challenge, atopy by allergen skin prick tests, and symptoms of asthma and aspects of lifestyle by questionnaire. Degree of Westernization was assessed from dietary habit categorized as exclusively Asian, mostly Asian, mixed, or exclusively English. The questionnaire was completed by 92% of children. The 308 white and 539 Asian children were of similar age (mean 9.3 yr versus 9.4 yr) with slightly more girls in the Asian group (52.6% versus 47.4%). The white children reported more wheeze within the past year (14.2% versus 8.8%, p = 0.01) but less were atopic (29.5% versus 38.4%, p = 0.01) with no significant difference in the prevalence of hyperreactivity (31.5% versus 29.6%, p = 0.57). Most Asians (97%) included Asian food in their diet; 35% ate a mostly Asian diet, 45% ate a mixed English and Asian diet, and 17% ate an exclusively Asian diet. After controlling for confounding factors whites were at higher risk of hyperreactivity and lower risk of atopy, and the risk of hyperreactivity and to a lesser extent atopy among Asian children was related to diet in a dose response fashion. The relative odds for hyperreactivity with an exclusively Asian, mostly Asian, or a mixed diet relative to an English diet were 0.31 (95% CI 0.15 to 0.62), 0.88 (0.56 to 1.37), and 0.99 (0.65 to 1.49), and for atopy were 1.22 (0.72 to 2.06), 1.47 (0.99 to 2.19), and 1.78 (1.23 to 2.58). These results suggest that diet or other closely related factors associated with Westernization are involved in the etiology of atopy and hyperreactivity.