Ability of 16 priority PAHs to be photocytotoxic to a cell line from the rainbow trout gill

Sixteen polycyclic aromatic hydrocarbons (PAHs) were screened for their ability to be photocytotoxic to a cell line from the rainbow trout gill, RTgill-W1. PAHs could be divided into one of three groups: incapable of being photocytotoxic, able to be both photocytotoxic and directly cytotoxic, or capable of being only photocytotoxic. Photocytotoxicity was distinct from direct cytotoxicity in that EC50 values were lower with the neutral red assay immediately after the PAH/UV treatment than with alamar Blue or CFDA-AM, indicating a more specific action on lysosomes. As well, in photocytotoxicity but not in direct cytotoxicity, the three assays showed increased impairment 24 h after treatment. Most PAHs were found to be strictly photocytotoxic; however, only six compounds were photocytotoxic at concentrations theoretically achievable in water. When photocytotoxic PAHs were ranked relative to fluoranthene to establish fluoranthene equivalent factors (FEFs), benzo[a]pyrene and benzo[g,h,i]perylene were found to be most potent. However, when the water solubility of each compound was taken into account in order to calculate the potential environmental photocytotoxic potency (PEPP), fluoranthene and pyrene appeared to have the most potential to impact fish through photocytotoxicity.     

Ability of 16 priority PAHs to be directly cytotoxic to a cell line from the rainbow trout gill

Sixteen polycyclic aromatic hydrocarbons (PAHs) were screened for their ability to be directly cytotoxic to a cell line from the rainbow trout gill, RTgill-W1. Exposure times of 2 h or less were sufficient for direct cytotoxicity to be detected, which appeared to be caused by a common mechanism, the general perturbation of membranes. This was judged by the similarity of results obtained for three fluorescent indicator dyes, alamar Blue, 5-carboxyfluorescein diacetate acetoxymethyl ester (CFDA-AM) and neutral red. Among the 16 PAHs tested, just two- and three-ring PAHs were found to be directly cytotoxic. These were naphthalene approximately = acenaphthylene approximately = acenaphthene > fluorene approximately = phenanthrene. The results suggest that water solubility and lipophilicity are the critical properties determining the direct cytotoxicity of PAHs and that they do so by influencing PAH accumulation in membranes. Only naphthalene was effective at concentrations well below its water solubility limit. Therefore, direct cytotoxicity is likely to be most environmentally relevant only with naphthalene.     

A digenean metacercaria from the reef coral, Porites compressa, experimentally identified as Podocotyloides stenometra

Feeding Porites compressa infected with a digenean metacercaria to the coral-feeding butterflyfish, Chaetodon multicinctus, established that the metacercaria was Podocotyloides stenometra. Those and field examinations finding a prevalence of 100% and an average intensity of infection of 6.5 worms/fish in 28 C. multicinctus off Kaneohe Bay, Oahu, Hawaii, established this fish as a definitive host. Plagioporus sp. of Cheng and Wong, 1974 is a synonym of P. stenometra.     

The effect of poly-L-lysine, amiloride and methyl-L-lysine on gill ion transport and permeability in the rainbow trout

The action of poly-L-lysine (PLL) on Na and Cl transport across freshwater fish gills was studied. Low concentration (10-6M) were added to the external medium for brief periods (1--5 min), then removed. During the next 20 min there was a rapid net loss of Na (117+/17 muEquiv[100 g]-1hr-1) and Cl (129+/17 muEquiv[100 g]-1hr-1). Both values are an order of magnitude larger than unidirectional effluxes in control fish. The efflux of both ions decreased to control values within 60 min after application and removal of PLL. In contrast, unidirectional influxes (JCl in and JNa in) were inhibited by about 40+ and showed no sign of returning to the original rates for 3 hr. Thus, PLL has two independent actions, causing a large increase in gill permeability which is reversible within an hour and apartial inhibition of influx which showed no sign of reversing for 2--3 hr. When PLL was applied for a longer period (60 min, the results were qualitatively similar but the permeability change was larger and persisted longer. These effects were compared with those of the small organic amines, amiloride and methyl-L-lysine. The latter inhibited JNa in, but there was not other similarity to PLL. Neither affected sodium efflux, not did they have any effect on Cl movements, in or out, across the gill. Inhibition of JNa in, was rapidly and completely reversible, and amiloride was shown to act by competing with Na for an entry site.     

The gill calcium transport cycle in rainbow trout is correlated with plasma levels of bioactive, not immunoreactive, stanniocalcin

The aims of this study were to determine the most appropriate duration for the measurement of the maximal accumulated O2 deficit (MAOD), which is analogous to the anaerobic capacity, to ascertain the effects of mass, fat free mass (FFM), leg volume (Vleg) and lower body volume (V1b) on anaerobic test performance, to examine the reproducibility for peak power output (Wpeak) or maximal anaerobic power using an air-braked cycle ergometer and to produce approximations for the percentages of aerobic and anaerobic metabolism during exercise of short duration but high intensity. A group of 12 endurance trained cyclists [mean age 25.1 (SD 4.6) years; mean body mass 73.43 (SD 7.12) kg; mean maximal oxygen consumption 5.12 (SD 0.35) l.min-1; mean body fat 12.5 (SD 4.1) %] accordingly performed four counterbalanced treatments of 45, 60, 75 and 90 s of maximal cycling on an air-braked ergometer. The mean O2 deficit of 3.52 l for the 45-s treatment was significantly less (P < 0.01) than those for the 60 (3.75 l), 75 (3.80 l) and 90-s (3.75 l) treatments. These data therefore indicate that in predominantly aerobically trained subjects the O2 deficit attains a plateau after 60 s of maximal cycling on an air-braked ergometer. Statistically significant interclass correlation coefficients (P < 0.05) between the anthropometric variables (mass, FFM, Vleg and Vlb) and Wpeak or maximal anaerobic power (0.624-0.748) and MAOD (ml) or anaerobic capacity (0.666-0.772) furthermore would suggest the relevance of taking into account muscle mass during anaerobic tests.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immunohistochemical detection of vitellogenin in male brown trout from Swiss rivers

The detection of vitellogenin, a yolk precursor protein, may serve as a biomarker for exposure to environmental oestrogens as its induction by xenobiotic oestrogens in the immature and male fish has been reported repeatedly. In the present work, juvenile brown trout were injected with oestradiol (5 microg g(-1) body weight oestradiol benzoate) in order to assess the induction and organ distribution of vitellogenin by means of immunohistochemistry. In addition, brown trout collected from Swiss rivers were analysed. Vitellogenin was detected in the oestradiol-injected juvenile trout but not in uninjected controls. The presence of vitellogenin was also demonstrated in a male and an immature feral brown trout from one of two locations downstream of three sewage treatment plants. In contrast, no positive staining was found in livers of trout upstream of the respective plants. The results demonstrate the suitability of immunohistochemistry for monitoring feral fish for the presence of vitellogenin production.     

A theoretical formulation for boundary friction in articular cartilage

In this paper we report the cloning and characterization of cDNA encoding a novel, short form of heparin-binding EGF-like growth factor (SF HB-EGF), and show expression of specific mRNA in various tissues and cell types. Our data suggest that SF HB-EGF mRNA is a product of alternative splicing. Like normal HB-EGF, SF HB-EGF contains the signal peptide, the propeptide, the heparin-binding domain and the first two conservative disulfide loops of the EGF unit. Instead of the third disulfide loop, the spacer, the transmembrane and the cytoplasmic domains, SF HB-EGF has a nine amino acid tail.     

Occurrence of Loma cf. salmonae in brook, brown and rainbow trout from Buford trout hatchery, Georgia, USA

We retrospectively analysed the long-term treatment results (median 8 years) of 31 patients with macroprolactinoma. 17 patients were treated by pituitary surgery (group 1) followed by long-term dopamine agonist therapy whereas 14 patients received long-term dopamine agonist therapy alone (group 2). 2 patients of group 1 and 1 patient of group 2 had external pituitary irradiation because of progressive disease. The two groups were comparable with respect to age, gender and initial prolactin (PRL) levels. At the end of the observation period dopamine agonist dosage could be reduced by 50% in group 1 and by 39.3% in group 2. Pituitary function did not change substantially during therapy. Complete remissions (no visible tumour in CT or MRI, normal PRL levels under current dopamine agonist medication) were achieved in 23.5% of group 1 vs. 21.4% of group 2, partial remissions (reduction of PRL and tumour size) in 35.3% vs. 64.3%, stable disease in 23.5% vs. 7.1% and progressive disease in 17.7% vs. 7.1% (differences not significant). Visual field defects showed 28.4% remissions (complete and partial) in group 1 versus 50% in group 2. Dopamine agonist therapy could be stopped definitively in only 1 patient of group 2 with an invasive macroprolactinoma. Initial surgical reduction of tumour load followed by medical therapy does not seem to guarantee a better long-term outcome than dopamine agonist therapy alone if the patient responds to and tolerates dopamine agonist therapy. Surgery should be reserved for non-responders, drug-intolerant or non-compliant patients, and for those with acute severe neurological compromise.     

A novel cartilage protein (CILP) present in the mid-zone of human articular cartilage increases with age

A novel, somewhat basic noncollagenous protein was purified from guanidine hydrochloride extracts of human articular cartilage using cesium chloride density gradient centrifugation, followed by ion-exchange chromatography at pH 5, and gel filtration on two serially coupled columns of Superose 6 and Superdex 200. The protein of 91.5 kDa contains a single polypeptide chain substituted with N-linked oligosaccharides. It appeared unique to cartilage as studied by enzyme-linked immunosorbent assay and immunoblots of various tissue extracts. Its concentration in articular cartilages showed some variability with age being lower in young individuals. It represents a chondrocyte product, since it is synthesized by articular chondrocytes in explant cultures. Interestingly, the distribution of the protein in the articular cartilage provides important information on the nature of chondrocytes at different compartments in the tissue. Thus, chondrocytes in the middle/deeper layers of the tissue in particular, appeared to have produced the protein and deposited it in the interterritorial matrix. The protein was neither seen in the superficial nor in the deepest regions of the articular cartilage. Based on its immunolocalization we have named this protein CILP (cartilage intermediate layer protein).     

A prion protein fragment primes type 1 astrocytes to proliferation signals from microglia

Giliosis is a hallmark of prion disease. A neurotoxic prion peptide (PrP106-126) induces astrocyte proliferation in the presence of microglia. This peptide also directly enhances microglial proliferation in culture. We have investigated this further to understand the method by which factors released by microglia and PrP106-126 work together to enhance astrocyte proliferation. PrP106-126 in the presence of microglia specifically enhanced type 1 astrocyte proliferation but not Type 2. Astrocytes that do not express the prion protein were more sensitive to oxidative stress and the toxicity of cytosine arabinoside. In the presence of cytosine arabinoside, PrP106-126 was toxic to pure astrocyte cultures. Using conditioned medium from microglia we have shown that PrPc-expressing astrocytes proliferate in response to factors released by microglia stimulated by granulocyte/macrophage colony-stimulating factor. This response is enhanced in the presence of PrP106-126. PrPc-deficient astrocytes do not show this response. These results suggest that astrocytes are primed by PrP106-126 to respond more to factors released by proliferating microglia. Astrocytes may proliferate in this system to escape entering the cell suicide pathway.     

A physiologically based pharmacokinetic and pharmacodynamic model for paraoxon in rainbow trout

Trout were exposed to an aqueous solution of 75 ng/ml paraoxon for 5 days at 12 degrees C. The relationships among paraoxon concentration in water and target organs, AChE inhibition, and carboxylesterase (CaE) detoxification of paraoxon were characterized quantitatively by development of a PBPK-PD model. The PKPD model structure consisted of brain, heart, liver, kidney, and remainder of the body, which were interconnected by blood circulation. The paraoxon tissue/blood partition coefficients were: plasma/water, 1.46; liver/plasma, 5.89; brain/plasma, 3.90; heart/plasma, 2.91; kidney/plasma, 0.45; and blood/plasma, 0.91. Turnover of AChE was characterized from a dose-response study, in which its zero-order synthesis rate and first-order degradation rate constant were determined in several tissues; for brain they were 7.67 pmol/min and 7.31 x 10(-5) hr(-1). The uptake and depuration clearances of paraoxon (Cl(u) = 0.651 and Cl(d) = 0.468 ml min(-1) g body wt(-1)) were determined using a compartmental model. During continuous water exposure to paraoxon, AChE activity in the tissues declined to new steady state values that were maintained by the synthesis of new AChE. CaE was shown by simulation to be an important pathway for detoxification of paraoxon.     

A new technique for nasal tip cartilage graft in primary rhinoplasty

Inadequate tip projection in traditional rhinoplasty led to postrhinoplasty deformity, which with time got worse. Sheen's vertical shield and Peck's horizontal multiple rectangular stacked cartilage graft have been used to reduce postrhinoplasty tip projection loss. The new two fixed column disk cartilage graft technique at the tip-defining points is designed to combine the advantages of both techniques. This technique can be used in both open and closed rhinoplasty, and the amount of projection can be adjusted to fit the dorsum with relative ease. This technique has been used in 38 primary rhinoplasties with satisfactory results.     

Allyl sulfides from garlic suppress the in vitro proliferation of human A549 lung tumor cells

This study was designed to evaluate the alterations in doppler derived coronary blood flow velocities and flow reserve following rotational ablation. Changes in doppler derived coronary blood flow velocity variables have been valuable in assessing the physiological outcome following coronary balloon angioplasty. Rotational ablation's mechanism of plaque removal could alter distal vascular bed characteristics, and, as a result, intracoronary blood flow velocities and the coronary flow reserve. A 12-MHz doppler guidewire recorded intracoronary phasic velocities and coronary flow reserve (as assessed by the hyperemic response to adenosine [12-18 mcg intracoronary]) in 28 patients, before and after rotational ablation of 30 lesions. Adjunctive balloon angioplasty was performed in 27 of 28 patients (96%). Rotational ablation and adjunctive balloon angioplasty successfully reduced the lesion diameter (87 +/- 9% to 14 +/- 11%; P < 0.001). A significant increase in the mean distal average peak velocity (25 +/- 13 cm/sec, before; 47 +/- 22 cm/sec, after; P < 0.001), and decrease in the proximal to distal average peak velocity ratio, (2.1 +/- 1.3; to 1.2 +/- 0.4; P = 0.002) was recorded. The mean distal diastolic to systolic velocity ratio (before, 1.4 +/- 0.7; after, 1.6 +/- 0.8; P = 0.44) and the coronary flow reserve (before, 1.6 +/- 0.6; after, 1.5 +/- 0.5; P = 0.34) did not increase despite increases in distal velocities, following successful intervention. Doppler derived distal coronary blood flow velocities increased following rotational ablation and adjunctive balloon angioplasty, with resolution of transstenotic velocity gradient. Changes in distal phasic velocity pattern and coronary flow reserve, immediately after the intervention, were not useful in the assessment of the functional outcome and may be related to abnormalities in distal vascular bed vasoreactivity produced by rotational ablation.     

Isolation of a spotted fever group rickettsia from the Pacific Coast tick, Ixodes pacificus, in Oregon

A rickettsia of the spotted fever group was isolated on three occasions from Ixodes pacificus in western Oregon. These isolations, and additional evidence furnished by complement fixation tests on guinea pigs inoculated with other Oregon ticks of this species, indicate that the association of this rickettsia with the Pacific Coast tick may be widespread. This is the first isolation of a spotted fever group rickettsia from I. pacificus. Because the Oregon isolates are mildly virulent for guinea pigs they resemble the Western U and Rickettsia montana strains of rickettsiae. However, preliminary evidence from cross-immunofluorescence tests of mouse antisera suggests the Tillamook and Grants Pass strains are antigenically different from all known spotted fever group agents.     

Identification of glycoproteins in goblet cells of epidermis and gill of plaice (Pleuronectes platessa L.), flounder (Platichthys flesus (L.)) and rainbow trout (Salmo gairdneri Richardson)

A quantitative analysis has been made of the glycoproteins present in the goblet cells of the epidermis, gill filaments and gill lamellae of three species of teleost fish. The glycoproteins have been identified by a combination of techniques, including the use of the enzyme sialidase followed by Alcian Blue staining, at PH 2.6 or 1.0, in combination with periodic acid-Schiff. The selected fish were representative of species living in marine, freshwater and estuarine environments. The range of glycoproteins identified in these fish was similar to that found in mammalian tissue in that both neutral and acid glycoproteins were present, the latter included both sialomucins sensitive and resistant to sialidase, and sulphomucin. A single goblet cell contained either neutral or acid glycoproteins alone or in combination. Only the epidermis of the plaice and rainbow trout contained uniform cell populations producing acid glycoproteins, the former sulphomucin and the latter mainly sialomucin. At each site in the flounder and in the gill epithelia of the plaice and rainbow trout, the goblet cell population was mixed, with cells producing each type of glycoprotein. The number of goblet cells producing each type of glycoprotein varied at each tissue site.     

A scanning electron microscopic study of the degenerative cartilage in patellar chondropathy

Patellar chondropathy as cartilage degeneration localized in patellar cartilage in young persons is characterized by cartilaginous changes, such as softening, swelling, and fissuring. With a view to structural characterization of early cartilaginous degeneration before erosion, the morphology of affected cartilage was studied under a scanning electron microscope. The surface network of cartilage constituting fibrils had an edematous change, presenting with fibrillation on the medial facet, whereas many fibrils of the central ridge had a collagen bundle, and fissuring of varying size was observed. It appeared that a mechanical force (shearing) acting on the site of the central ridge was associated with the formation of a collagen bundle and its destruction. On the lateral facet, fibrils were arranged perpendicular to the joint surface; the superficial layer of fibrils was worn by hyper-pressure acting on the lateral facet. On the fractured surface, the coarseness of collagen fibrils showed changes that varied with the site and stage of cartilage degeneration. Frequent changes were signs of fibril loosening (coarsening), such as reduction in fibril density (i.e., edematous change), collagen fibril aggregation, and fissuring, and longitudinal restructuring of fibrils. The patellar cartilage in the patients of this series showed a structure adapted to the mechanical force. The initial structural changes of cartilage consisted of collagen fibril aggregation and reduction in fibril density. These changes give rise to matrix rarefaction, which in turn causes cartilage degeneration to progress. These changes were concurrent in both the superficial and middle layers and were not localized as basal degeneration.     

Yeasts isolated from the intestine of rainbow trout adhere to and grow in intestinal mucus

The hyaline layer of echinoderm embryos is an extraembryonic matrix that functions as a substrate for cell adhesion through early development. The major constituent of the hyaline layer is the protein hyalin, a fibrillar glycoprotein of approximately 330 kDa that multimerizes in the presence of calcium. Here we provide a molecular characterization of hyalin and identify a region of the protein that is important for its function in cell adhesion. Partial hyalin cDNAs were identified from two sea urchin species, Strongylocentrotus purpuratus and Lytechinus variegatus, by screening expression libraries with monoclonal antibodies to hyalin. The cDNAs each encode a tandemly arranged series of conserved repeats averaging 84 amino acids. These hyalin repeats are as similar between the two species as they are to repeats within each species, suggesting a strong functional conservation. Analysis of this repeat shows that it is a unique sequence within the GenBank database with only weak similarity to mucoid protein sequences. The hyalin mRNA is approximately 12 kb in length and is present in developing oocytes coincident with the appearance of cortical granules, the vesicle in which the hyalin protein is specifically packaged. The mRNA is present throughout oogenesis but is rapidly lost at oocyte maturation so that eggs and early embryos have no detectable hyalin mRNA. The hyalin protein, however, remains at relatively constant levels throughout development. Thus, all the hyalin protein present during early development, when no RNA is detectable, is maternally derived and exocytosed from cortical granules at fertilization. Hyalin mRNA reaccumulates in embryos beginning at the mesenchyme blastula stage; a RNA gel blot and in situ hybridization analysis of gastrulae and larvae shows a progressive confinement of hyalin mRNA to the aboral ectoderm. Recombinant hyalin containing the tandem repeat region of the protein was expressed in bacteria and is shown to serve as an adhesive substrate, almost equal to that of native hyalin, in cell adhesion assays. This adhesive activity is partially blocked by dilute hyalin monoclonal antibody Tg-HYL to the same extent as that for native hyalin. Thus, this hyalin repeat region appears to contain the ligand for the hyalin cell surface receptor. These data help explain some of the classic functions ascribed to the hyalin protein in early development and now enable investigators to focus on the mechanisms of cell interactions with the hyaline layer.     

Hairs causing pulmonary emboli. A rare complication in long-term intravenous studies in dogs

From a study of 320 individuals superoxide dismutase (SOD) variants are shown to be present in Iraq, and it is suggested that this genetic system may be useful for the study of historical population movement in the Middle East. The esterase D gene frequencies show no significant heterogeneity among the different regions of the country.