Development of two novel CYP-antibodies and their use in a PCB exposure experiment with Mytilus edulis.

In an attempt to learn more about the cytochrome P450 (CYP) system of mussels, we used protein databases and alignment software to extract highly conserved CYP sequences. From these alignments synthetic peptides were produced and used for rabbit immunisation, which yielded polyclonal antibodies against the CYP families 2 and 4. The antibodies were evaluated with Western Blot and ELISA assays, using digestive gland microsomal samples from the mussel Mytilus edulis. Western Blots revealed immunoreactions for both antibodies. The anti-CYP2 sequence rendered one major immunopositive protein of approximately 49 kDa size, and weak signals for proteins of approximately 41 and 56 kDa size. The anti-CYP4 sequence rendered two major bands of approximately 56 and 59 kDa size, and also a weak immunoreaction with a protein of approximately 43 kDa size. ELISA rendered only weak signals even with a 1:50 dilution of IgG-purified serum. A 10-day exposure to Aroclor 1254 did not appear to affect any of the immunopositive proteins, while total PCBs in soft bodies increased from 14-40 ng/g DW in controls to 373-638 ng/g DW in exposed mussels.

Dietary vitamin C, immunity and disease resistance in Atlantic salmon (Salmo salar).

Presmolt Atlantic salmon were fed a fish meal based experimental diet supplemented with graded levels of ascorbate-2-monophosphate (AP), equivalent to 40, 400, 2000 and 4000 mg ascorbic acid (AA)/kg for 6 months prior to a bacterial challenge experiment. The liver AA concentration reflected the dietary intake of AP, but not linearly. Growth, hematology and acid phosphatase activity in zymosan stimulated macrophages were not affected by dietary AP. Serum hemolytic complement activity was higher in fish fed the highest AP level, but the variation was not significant. Production of specific antibodies was significantly higher in fish fed the highest AP level 11 and 17 weeks after vaccination. Bacterial challenge with Aeromonas salmonicida showed increased survival in the 4000 AP group. Lysozyme activity in headkidney and serum complement activity and serum iron in fish surviving the challenge were higher in the 4000 AP group, indicating important roles of vitamin C on lysozyme, complement and iron in non-specific disease resistance. The results indicate that high dietary levels of AP favourably affect health in Atlantic salmon. Disease resistance was, however, not correlated with the AA status in the liver.

The minimum dietary requirement of vitamin C in Atlantic salmon (Salmo salar) fry using Ca ascorbate-2-monophosphate as dietary source.

The minimum dietary vitamin C requirement for optimal growth and normal development in Atlantic salmon (Salmo salar) fry at the onset of feeding was studied, using Ca ascorbate-2-monophosphate (AP) as dietary source. The requirement was established by means of a feeding study lasting for 23 weeks from the beginning of feeding. The practical diets used were supplemented with AP at levels of 0, 10, 20, 40, 80 and 160 mg ascorbic acid (AA) equivalents/kg. Growth, mortality, hydroxyproline content in skin and backbone, and AA in liver were recorded to evaluate the results. The results suggest that the minimum dietary requirement for optimal growth and normal development is in the range of 10-20 mg AA equivalents/kg dry diet during the period studied.

Cardiac myxoma as the cause of death in an infant.

Cardiac myxoma is a rare disease most frequently encountered in adults, usually localized in the left atrium. It rarely occurs in infants. We report a 3 1/2 month old girl whose sudden death was caused by a large atrial myxoma, completely occluding the tricuspidal orifice. The importance of possible prodromal symptoms suggesting a cardiac disease is stated, and the role of echocardiography as an excellent examination technique is discussed.

Ascorbate-2-sulfate as a dietary vitamin C source for atlantic salmon (Salmo salar): 1. Growth, bioactivity, haematology and humoral immune response.

The present experiment shows that ascorbate-2-sulfate (AS) is not equivalent to ascorbic acid (AA) as a dietary vitamin C source for Atlantic salmon (Salmo salar). Within reasonable feed supplemental levels AS does not provide the tissues with adequate supplies of vitamin C to secure optimal physiological functions as demonstrated by biochemical and haematological analyses.AS could not be detected in the liver of fish fed either AA or AS, nor in vitamin C - deprived salmon, suggesting that AS is not the natural storage form of vitamin C in this species.There were no significant differences in antibody production against a soluble artificial antigen (NIP11-LPH) in fish fed 500 and 5000 mg AA/Kg dry diet or equivalent amounts of AS during a period of six weeks at a water temperature of 7.2°C.

Ascorbate-2-sulfate as a dietary vitamin C source for Atlantic salmon (Salmo salar): 2. Effects of dietary levels and immunization on the metabolism of trace elements.

Atlantic salmon fingerlings were fed a vitamin C deficient diet for four months. The fish were then provided a dry, practical fishmeal based diet supplemented with 0, 500 or 5000 mg vitamin C/kg as L-ascorbic acid or equivalent amounts of ascorbate-2-sulfate. After six weeks on these diets ten fish in each group were injected with a soluble antigen (NIP11-LPH). Six weeks thereafter blood, liver, kidney, spleen and vertebrae were examined for trace elements. The livers were also analysed for metallothionein.The vitamin C deficient fish were anemic despite the significantly elevated iron concentrations in the liver. Vitamin C had no positive effect in lowering tissue levels of cadmium. The highest level of dietary vitamin C given as ascorbic acid reduced the liver selenium concentrations. In response to antigen injection, the fish in all groups showed increased levels of hepatic metallothionein, copper, zinc and cadmium, while hepatic selenium and iron levels were less affected. The elemental composition in other organs was affected by the antigen injection to a minor extent.

Separation of mouse epidermal basal and differentiating cells for microflow fluorometric measurements: a methodologic study.

The DNA content of lymphocytes and of basal cells from normal hairless mouse epidermis was measured by microflow fluorometry (MFF). To obtain a relatively pure suspension of epidermal basal cells a combined mechanical and enzymatic method was used. The admixture of differentiating cells into the basal cell fraction after cell separation was 13%. The results were compared with those obtained with conventional Feulgen microspectrophotometry applied to basal cells and dermal lymphocytes in histologic sections. The results from both cytophotometric methods were in good agreement and clearly demonstrated the improved resolution obtained by using microflow fluorometry. When the lymphocytes were not treated with pepsin before being stained with ethidium bromide for MFF, the modal DNA value was consistently below that of the basal cells from the same specimen. Pepsin treatment of lymphocytes, however, increased their fluorescence intensity to the value of epidermal basal cells. The modal DNA value of Feulgen-stained dermal lymphocytes in histologic sections was consistently below that of epidermal basal cells from the same section. The advantage of pepsin treatment for obtaining higher resolution of DNA measurements of basal and differentiating epidermal cells and of lymphocytes was evaluated. The cell cycle distribution of basal cells from epidermis in different states of proliferative activity was determined. Changes in the proportion of cells in S phase were parallel to changes in the 3H-Tdr labeling index.

Polycythemia vera treated with -32p and myleran: development of chronic granulocytic leukemia with chromosomal abnormalities in one patient.

Chronic granulocytic leukemia developed in a 59-year-old woman who had previously received a total of 21 mCi -32P for polycythemia vera. She was treated with Myleran (busulphan) for her chronic granulocytic leukemia. Cytogenic studied revealed deletion of chromosomes No. 8 and 12, and translocation between 1 and 8. The patient also developed a severe antoimmune hemolytic anemia, for which she received prednisone treatment. She died with a perforated stomach ulcer.