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Biomedical subjects

R Johansen

Publications and source records attributed to R Johansen.

At least 19 recordsLinked to original sources

Guidelines for health and welfare monitoring of fish used in research.

The aim of this paper is to provide background material necessary for the development of international guidelines for the health and welfare monitoring of fish used in research. It provides an overview of present guidelines and discusses why more detailed and species-specific guidelines are needed. A major issue within fish research is to document the situation today and point out areas where improvements are needed.

Animal Welfare↗

A sequential study of pathological findings in Atlantic halibut, Hippoglossus hippoglossus (L), throughout one year after an acute outbreak of viral encephalopathy and retinopathy.

Following a natural outbreak of viral encephalopathy and retinopathy (VER) at a commercial farm in Norway, surviving Atlantic halibut, Hippoglossus hippoglossus, were sequentially studied for distribution of nodavirus, immune response and histopathology over 1 year. Typical clinical signs and histopathology of VER were observed during the acute stage of the disease. Most of the surviving fish became subclinical carriers of nodavirus with clusters of nodavirus-containing cells in the central nervous system. Four random samplings of presumably healthy fish were performed from two fish groups, with low and high growth rates respectively, over a 7-month period. Immunohistochemical (IHC) examination revealed a higher number of nodavirus-positive cells in fish with a low growth rate than in fish with a high growth rate. All IHC positive fish were also reverse transcriptase polymerase chain reaction (RT-PCR) positive for nodavirus and for nodavirus antibodies detected by enzyme-linked immunosorbent assay (ELISA) at all sampling points. The percentage of PCR- and ELISA-positive fish remained high throughout the year, while the number of IHC-positive fish decreased, especially in the group with a high growth rate. Several other histopathological changes were observed, including pericarditis, steatitis, changes in liver and kidney, and necrosis of the intestinal wall. None of these findings seemed to be related to the nodavirus infection. Nodavirus was reisolated in cell culture from subclinically infected fish one year after the acute VER outbreak, which indicates that the virus was still infectious.

Animals↗

Characterization of nodavirus and viral encephalopathy and retinopathy in farmed turbot, Scophthalmus maximus (L.).

An outbreak of nodavirus infection in turbot larvae is described with respect to histopathology, immunohistochemistry, cell culture cultivation, RT-PCR amplification and sequence analysis of the capsid protein gene RNA2. Affected turbot developed classical signs of viral encephalopathy and retinopathy (VER) with abnormal swimming behaviour and high mortality levels. In the acute stage of infection, light microscopy revealed vacuolation of the central nervous system (CNS), with positive immunohistochemical staining for nodavirus. Later in the infection, CNS lesions appeared more chronic and contained clusters of cells immunopositive for nodavirus. Bacterial overgrowth in the intestines of the fish may have provoked or influenced the course of the nodavirus infection. We were unable to propagate the virus in cell culture. While RT-PCR using primers designed to detect Atlantic halibut nodavirus gave negative results, further testing with primers complementary to a more conserved region of RNA2 resulted in amplification of a product of the expected size. The entire RNA2 segment was cloned and sequenced. Sequence alignment showed that the turbot nodavirus (TNV) was different from previously described fish nodaviruses. In addition, phylogenetic analysis based on an 823 nt region of the sequence indicated that TNV clustered outside the four established fish nodavirus genotypes, suggesting a fifth genotype within the betanodaviruses.

Amino Acid Sequence↗

Acute and persistent experimental nodavirus infection in spotted wolffish Anarhichas minor.

Spotted wolffish Anarhichas minor (approx. 0.7 g) were found to be susceptible to infection with a nodavirus isolated from Atlantic halibut (AHNV) by bath-challenge. During the acute stage of infection, 4 to 8 wk post-challenge, viral encephalopathy and retinopathy (VER) were diagnosed by histopathology, immunohistochemistry (IHC) and reverse transcriptase-polymerase chain reaction (RT-PCR). Accumulated mortality was 52% in the challenged group. The surviving fish were sampled 16 wk post-challenge, by which time they had grown to approximately 17 g. No clinical signs of VER were observed in these fish. RT-PCR examination revealed the presence of nodavirus in several organs of the survivors, but no immunopositive cells were detected by IHC. Nodavirus was reisolated from fish at the last sampling in SSN-1 cells, showing that nodavirus retains virulence in persistently infected wolffish for at least 16 wk post-bath-challenge.

Animals↗

Experimental infection of Atlantic halibut Hippoglossus hippoglossus with nodavirus: tissue distribution and immune response.

Atlantic halibut Hippoglossus hippoglossus, age 8 mo and weighing 20 g, were challenged by either intraperitoneal injection (i.p.) or by bath exposure using nodavirus isolated from Atlantic halibut. Fish were sampled at intervals over a 41 d period, starting on Day 5 post-challenge. Although no clinical disease or mortality was recorded, the data show that nodavirus did successfully propagate in i.p.-challenged fish. Using conventional end-point reverse transcription (RT)-PCR, nodavirus was detected in the kidney of all examined i.p.-challenged fish, and further in the head, heart, liver and posterior intestine of most of these individuals. Quantitative real-time RT-PCR revealed that the amount of virus in head samples from the i.p.-challenged group increased during the experiment. The presence of nodavirus in nervous tissue of i.p.-challenged fish was detected by immunohistochemistry from Day 13 post-challenge. In the retina, virus positive cells were found adjacent to the circumferential germinal zone at the ciliary margin towards the iris. In the brain, a few positive cells were detected in the tectum opticum. An ELISA was developed to detect anti-nodavirus activity in plasma. The method included an optimized coating procedure, which allowed the use of non-purified nodavirus as the coating antigen in a simple indirect ELISA. An anti-nodavirus antibody response was detected from Day 19 post-challenge in i.p.-challenged fish, while a response was not detected in the bath-challenged or control fish. This experiment demonstrates a subclinical nodavirus infection in Atlantic halibut at a post-juvenile stage induced by i.p. injection of virus.

Animals↗

Pericarditis and myocarditis in farmed Atlantic halibut Hippoglossus hippoglossus.

Farming of Atlantic halibut Hippoglossus hippoglossus is a growing industry in Norway. This study comprises the first description of heart morphology in fish from one Atlantic halibut farm. Fish with no clinical signs of disease were randomly sampled (8 to 12 mo old, weight 1 to 50 g). The hearts were studied by light and electron microscopy and samples were taken from the kidney and pericardial cavity for bacteriological examination. Slight to severe changes were found in most hearts at all samplings. The most serious pericardial and myocardial changes were seen in larger fish (15 to 50 g). In some of the hearts the affected pericardium formed multifocal protrusions. Eight fish had an adhesion between the apex of the ventricle and the parietal wall of the pericardial cavity. Results of bacteriology and immunohistochemistry investigations for common disease agents were negative.

Animals↗

[Nerve biopsy].

Nerve biopsy, if undertaken at centres with appropriate expertise, may be a valuable diagnostic procedure in selected patients with peripheral neuropathy. Sural nerve biopsy and skin biopsy for evaluation of epidermal nerves are described. Indications, surgical procedure, preparation and transport are reviewed. Usually the patient has to be admitted to a hospital with the necessary resources for a comprehensive battery of nerve tissue processing, including teasing and electron microscopical examination, and other investigative procedures. Another option, when practically and economically feasible, is to obtain sural nerve biopsy from several (2-4) patients performed by a skilled surgeon at the local (central) hospital. A technician from a neuromuscular centre or laboratory of neuropathology should be at the site to provide immediate and proper preparation and transport of the tissue samples. An example of peripheral nerve vasculitis with teased nerve fibers in early and late axonal degeneration is presented as an illustration. Sural nerve biopsy is useful in selected cases, particularly in order to demonstrate peripheral nerve vasculitis. For detection of small fiber neuropathy in the investigation of painful neuropathies, skin biopsy for evaluation of epidermal nerves may be an appropriate alternative.

Biopsy↗

Reactivity of leg alignment to articular pressure testing: evaluation of a diagnostic test using a randomized crossover clinical trial approach.

OBJECTIVE: A study was undertaken to assess the reliability of detecting leg alignment changes (reactivity) and to determine if the observed leg alignment reactivity can be attributed to a rotatory articular pressure challenge. DESIGN: Prospective double-blind crossover trial of a diagnostic test. SETTING: Laboratory: Center for Technique Research. PARTICIPANTS: Forty-two chiropractic college students, faculty and staff. INTERVENTIONS: A standardized force of 2 or 3 kg was applied with a 1 cm2 rubber-tipped pressure algometer on the lateral aspect of the T3-T7 spinous processes and the posterior aspect of the lateral masses of C1. MAIN OUTCOME MEASURES: Leg alignment reactivity: an increase in leg alignment discrepancy (yes or no) following a diagnostic intervention. RESULTS: The reliability for detecting leg alignment reactivity was poor: on average, Kappa = 0.05 in the thoracics and 0.06 at C1. On average, the attributable risk of leg alignment reactivity (pressure test risk--sham test risk) was less than 4%. In many cases, the sham rate was greater than the pressure test rate. CONCLUSIONS: For the population investigated, leg alignment reactivity to rotatory pressure testing can, in the majority of cases, be attributable to background noise. This procedure was not found to be viable for identifying vertebrae for adjustment. Further research with different subject populations, regions of investigation, leg alignment measurement techniques and vertebral challenge techniques are indicated.

Adult↗

Responsiveness of leg alignment changes associated with articular pressure testing to spinal manipulation: the use of a randomized clinical trial design to evaluate a diagnostic test with a dichotomous outcome.

OBJECTIVE: A study was undertaken to assess the stability of leg alignment reaction to a pressure challenge and its responsiveness to an adjustive intervention. DESIGN: Prospective, double-blind clinical trial of a diagnostic test. SETTING: Laboratory: Center for Technique Research. PARTICIPANTS: Forty-two chiropractic college students, faculty and staff. INTERVENTIONS: A high-velocity, low-amplitude, short lever adjustment of a single vertebra from among C1 and T3-T7; or a sham adjustment similar to a manual diagnostic pressure test at C1, T3-T7 or T9-T10. MAIN OUTCOME MEASURES: Leg alignment reactivity: An increase in leg alignment discrepancy (yes or no) following a metered pressure challenge to a vertebra. RESULTS: On average, stability was poor at T3-T7 (Kappa = 0.04), moderate at C1 (K = 0.47), and fair for sham pressure tests (K = 0.30). Responsiveness: The proportion of positive baseline leg alignment reactions that responded (became negative) to sham adjustment was 95% at T3-T7 and 55% at C1. Further analysis was untenable since too few vertebrae were implicated for an adjustment. CONCLUSIONS: For the population investigated, the majority of the responsiveness of the leg alignment diagnostic test to a rotatory adjustment appears to be a diagnostic illusion (i.e., background noise unrelated to a treatment intervention). Further research with different subject populations, regions of investigation, leg alignment measurement techniques and vertebral challenge techniques are indicated.

Adult↗

Artefacts and morphological changes during chemical fixation.

The normally 'condensed' (darkly stained) chromosomes of dinoflagellates decondense by swelling. This occurs in an increasing number of cells when the concentration of added OsO4 is decreased. With different fixatives other types of disintegration can be observed, which vary with the concentration. With cryofixation and freeze-substitution the chromosomes are most 'condensed'. Escherichia coli infected with bacteriophage T4, with or without active lysozyme production, were studied by optical densitometry for partial lysis and by light and electron microscopy for observing swelling. When active lysozyme is present some of the acrolein (2.5%)-glutaraldehyde (2%)-fixed cells swell at 0 degrees C, but do not in the absence of lysozyme nor when fixed at room temperature. If OsO4 is added at concentrations < or = 0.5%, partial lysis occurs when lysozyme is present. The optical density decreases, the cells lose some matter and swell slightly. The corresponding electron micrographs show gap formation by curdling and/or a decreased concentration of the cytoplasm which reveals certain phage-related particles.

Animals↗

Coralline shape of the bacterial nucleoid after cryofixation.

A new procedure of immunostaining sections of cryofixed and freeze-substituted Escherichia coli shows that DNA extends from its bulk into small ribosome-free spaces throughout the cytoplasm, resulting in a coralline-shaped nucleoid. Low-resolution imaging of a bacterium reconstructed from serial sections demonstrated that the small excrescencies are not resolved. The resulting photograph shows the same features as phase-contrast light micrographs.

Chromatin↗

Effect of cooling on the intracellular concentrations of Na+, K+ and Cl- in cultured human endothelial cells.

Cooling is accepted as a practical way of lowering cell metabolism in vein grafts during coronary by-pass surgery. We have previously shown that low temperature causes endothelial cells to become detached, both in in vitro and in vein graft. In this study we have looked at the effect of cold on the concentrations of intra- and extracellular electrolytes. Human endothelial cells were grown on titanium grids for electron microscopy. The cells were incubated for 30 min at 37 degrees, 20 degrees, and 4 degrees C with cell culture medium containing human serum, and at 20 degrees and 4 degrees C with heparinized sodium acetate solution with serum, frequently used for flushing and distending vein grafts. Freeze-dried cells were then subjected to elemental X-ray microanalysis. The ambient fluid was analysed by flame photometry. At 20 degrees and 4 degrees C, intracellular concentration of sodium increased, and potassium decreased, compared with controls (37 degrees C). The changes in sodium concentrations were aggravated when cell culture medium was replaced by heparinized sodium acetate. The intracellular chloride concentration did not change when cells were stored in cold cell culture medium. The extracellular concentration of potassium increased with increasing incubation time at 4 degrees C. The connection between these findings and cell detachment is discussed.

Chlorides↗

Dextran-induced lowering of parameters of the kallikrein-kinin system in rat plasma.

Pretreatment of rats with tranexamic acid inhibited the rapid lowering of the plasma levels of acetone/kaolin-activated prekallikrein proactivator and prekallikrein caused by intravenous injection of dextran, but did not inhibit the reduction in the level of plasminogen, and potentiated the lowering of high molecular weight kininogen. By acetone/kaolin activation of normal rat plasma a mixture of surface-bound factor XIIa and unbound XIIf was obtained, and a BAEe-esterase (MW about 47,000) possessing weak kininogenase activity was present in addition to kallikrein. In activated plasma from dextran-treated rats the cleavage of XIIa was strongly reduced, and the second esterase was almost absent. It is suggested that dextran induces the loss of a plasma factor which is important for the cleavage of factor XIIa in the adopted procedure. This factor was not high molecular weight kininogen, and the lowering of plasminogen was too small to account for the reduction in PKA-activity.

Animals↗

A family systems approach to home dialysis.

Dramatic increases in the cost of the end stage renal disease program are producing an emphasis on home dialysis with the patient and his family carrying the responsibility for this treatment. This requires an adjustment of the family's emotional system and role assignment to sustain members through the continued stress of chronic catastrophic illness. We are suggesting a model of intervention in which family members are encouraged to recognize stress caused by one member's illness and to make necessary changes in the family functioning.

Adult↗