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Biomedical subjects

F L Harrison

Publications and source records attributed to F L Harrison.

At least 19 recordsLinked to original sources

Effects of chronic irradiation on the reproductive success of the polychaete worm, Neanthes arenaceodentata.

Effects of lifetime exposure to chronic irradiation on reproductive success were assessed for laboratory populations of Neanthes arenaceodentata. Exposure was initiated upon the spawning of the parental (P1) female and was terminated upon spawning of the first filial (F1) generation female; broods from the F1 pairs were sacrificed before hatching occurred. Groups of worms in the experiments received either no radiation (controls) or 0.19, 2.1 or 17 mGy h-1. The total dose received was either background or approximately 0.55, 6.5 or 54 Gy, respectively. The mean number of embryos in the broods from the F1 females exposed to 17 mGy h-1 was statistically significantly different from the mean number of embryos from control females; however, the mean number of embryos in the broods from the F1 females exposed to 0.19 and 2.1 mGy h-1 was not significantly different from the mean number from control females. For all the radiation-exposed groups, there was a statistically significant reduction in the number and percentage of live embryos in the broods from the F1 pairs as well as a statistically significant increase in the numbers and percentages of abnormal embryos. Results on embryo abnormalities and mortalities indicate that dominant- and recessive-lethal mutations were most likely induced in the germ cells and that these mutations had an adverse effect on reproductive success by affecting the survival of early-life stages. Except for pairs exposed to 17 mGy h-1, there was no evidence of gamete killing or reduced fertilization success, because the number of developing embryos in the broods did not decrease with increased dose. Data for the estimated hatch number and actual hatch number indicated that doses as low as 0.19 mGy h-1 can reduce significantly the number of larvae that hatch when lifetime doses are given.

Animals↗

Effects of acute irradiation on reproductive success of the polychaete worm, Neanthes arenaceodentata.

Effects of acute irradiation on the reproductive success of a relatively low-fecundity species were investigated by exposing pairs of female and male polychaete worms (Neanthes arenaceodentata) to either no radiation (controls) or 0.5, 1.0, 2.0, 5.0, 10 or 50 Gy of acute irradiation (5 Gy min-1) at the time when oocytes were visible in the female. The broods from the pairs were sacrificed before hatching occurred, and information was obtained on the number in the brood, the number of normal and abnormal embryos, and the number of embryos that were living, dying and dead. Developing gametes of N. arenaceodentata appeared to be sensitive to acute irradiation. There was a significant reduction in the percentage of live embryos in the broods from pairs receiving doses as low as 0.5 Gy, which is lower than the lowest dose at which effects in invertebrates have been reported previously. This was most likely due to the induction of lethal mutations in the developing gametes, which affected the survival of embryos in early stages of life. Except for those pairs receiving 10 or 50 Gy, there was no evidence of decreased fertility or fecundity or of reduced fertilization success; the number of embryos in the broods from only these irradiated groups was significantly different from the controls.

Animals↗

The 14 kDa beta-galactoside binding lectin in myoblast and myotube cultures: localization by confocal microscopy.

The endogenous 14 kDa beta-galactoside-specific lectin has been localised in myoblasts and myotubes by indirect immunofluorescence using confocal microscopy. The antibodies used in these experiments suggest that in myoblast cultures the lectin is abundant on the cell surface and concentrated at ruffled edges of migrating cells. The quantity and distribution of cell surface lectin is independent of the nature of the substratum and undiminished by brief trypsinisation or lactose-washing of the cell suspension. The lectin is also abundant intracellularly, apparently relatively free in the cytoplasm until the cells approach confluency, when the lectin is concentrated into particular areas of cytoplasm resulting in a 'patchy' appearance of stained cells. After fusion to form multinucleate myotubes, intracellular lectin is less abundant and concentrated at the periphery of myotubes, from where lectin appears to be released in vesicles packed with lectin which 'bud off' from the myotube surface. In recolonising cultures, lectin is apparently more abundant and lectin-packed vesicles can also be seen 'budding off' from myoblasts. In differentiated cultures, extracellular lectin is detected both on the myotube surface and in fibrillar arrays of extracellular material.

3T3 Cells↗

The sugared path to a normal lung: soluble beta-galactoside-specific lectins.

The diverse functions of a family of soluble beta-galactoside-binding proteins in lung are discussed. The smallest and most abundant member, a 14-kDa lectin, appears to be a negative modulator of cell growth. Larger members of this family include RL29, a protein involved in the regulation of cytoplasmic RNA levels, and one subunit of the cell surface elastin receptor involved in the regulation of elastic fibrillogenesis. The structure and function of other members of this family remain to be elucidated, but probably homologous carboxy-terminal domains will specify the galactose-binding function and varied amino-terminal domains will specify the diverse biological functions. Although currently these functions are being described at the cellular level, these beta-galactoside-binding proteins appear to have important roles in regulating lung physiology.

Animals↗

The sequence of the mouse 14 kDa beta-galactoside-binding lectin and evidence for its synthesis on free cytoplasmic ribosomes.

The partial amino acid sequence of the mouse 14 kDa beta-galactoside-binding lectin has been deduced from cDNA clones corresponding to 86% of the coding sequence and extending to the polyadenylation signal. The deduced amino acid sequence for the murine lectin shows 94% identity with the rat, 89% with human, 86% with bovine and 46% with the chicken 14 kDa lectins. A cDNA probe has been used to analyse genomic DNA and identify a single mRNA of approx. 570 bp in 3T3 fibroblasts, murine erythroleukaemia cells and the murine basement-membrane-secreting Engelbreth-Holm-Swarm tumour. Analysis of free and bound polyribosomes has shown that the lectin message is translated on free cytoplasmic ribosomes.

Amino Acid Sequence↗

The bioavailability of sediment-sorbed chlorobenzenes to larvae of the midge, Chironomus decorus.

Larval stages of the midge, Chironomus decorus, were used to define the bioaccumulation of sediment-sorbed mono-, di-, tri-, and hexachlorobenzene. Larvae were exposed to high- and low-organic-content sediments that had been equilibrated with individual radiolabeled chlorobenzenes prior to testing. Equilibrium or nonequilibrium aqueous concentrations of the volatile test chemicals were flowed through sealed chambers in a sediment-water exposure system. The uptake of chlorobenzenes by midge larvae was rapid for all compounds tested, and apparent steady-state conditions were reached within 48 hr of exposure. Bioconcentration factors for the accumulation of chlorobenzenes from sediments and from interstitial and overlying waters were related to the octanol/water partition coefficients of the compounds. Because the diffusion of chlorobenzenes to overlying water during nonequilibrium flow through conditions was very slow, bioaccumulation was dependent on the concentration of the chemicals in interstitial water. These results show how benthic organisms may be able to accumulate significant levels of chlorinated aromatic compounds from ecosystems where their concentrations in the water column are relatively low.

Animals↗

Radiation-induced micronuclei in peripheral erythrocytes of Rana catesbeiana: an aquatic animal model for in vivo genotoxicity studies.

An in vivo micronucleus assay for peripheral erythrocytes of Rana catesbeiana tadpoles was developed and evaluated. The assay was used to determine the spontaneous frequency of micronuclei in circulating erythrocytes in tadpoles from two different populations, to define the time from administering the clastogen to the maximum micronucleus frequency in peripheral erythrocytes, and to determine the response to radiation. The spontaneous frequency of micronuclei in circulating erythrocytes of early-stage tadpoles was low (3.6 +/- 2.8 micronuclei per 1,000 erythrocytes, MN o/oo), but higher than that of late-stage tadpoles (1.7 +/- 0.7 MN o/oo). The time from the exposure of early-stage tadpoles to radiation (2.1 Gy) to the maximum micronucleus frequency was about 2 wk. The increase in frequency of micronuclei in peripheral erythrocytes of late-stage tadpoles receiving doses ranging from 0.5 to 3.0 Gy was linear with dose; a 3-fold increase was obtained with a dose of 3.0 Gy. The spontaneous frequency of micronuclei in erythrocytes and the increase in frequency induced by radiation appeared to differ in tadpoles from different populations. Quantification of micronuclei in the peripheral erythrocytes of R catesbeiana tadpoles provides a promising whole-animal system for studies of genotoxicity in aquatic environments.

Animals↗

Distribution of an endogenous beta-galactoside-specific lectin during foetal and neonatal rabbit development.

The tissue concentration of an endogenous beta-galactoside-specific lectin is developmentally regulated in rabbit tissues during foetal and neonatal development. Immunoassay data and localization studies both indicate that the lectin is particularly associated with late embryonic and early postnatal development. No lectin could be detected in embryos prior to 21 days of gestation, whereas many tissues showed an increase in lectin around the time of birth. It is likely that in most tissues the high concentrations of lectin are largely due to increased synthesis by fibroblasts, which are particularly abundant and active in connective tissue during the extensive tissue reorganization taking place at this time. The lectin appears to be synthesized by other differentiated cell types also, notably myoblasts, alveolar cells and erythroblasts. The peak of lectin concentration seen in foetal liver probably reflects lectin associated with foetal erythropoiesis. The rabbit lectin has a low specific activity but its tissue concentration is correspondingly higher than lectin concentrations in other mammals. This conservation of total lectin activity suggests a fundamental role for the lectin dependent upon its saccharide binding activity. This and other indirect evidence suggests that these lectins are involved in the synthesis, secretion or organization of extracellular matrix components.

Animals↗

Intra- and extracellular distribution of an endogenous lectin during erythropoiesis.

An endogenous beta-galactoside-specific lectin has previously been isolated from rabbit bone marrow. The quantification of extracted lectin now indicates that approximately 75% of the lectin is intracellular in marrow. Indirect immunofluorescence studies show the extracellular lectin is associated with the erythroblast cell surface and is also found in some acellular areas of the marrow stroma. At enucleation, lectin surrounds the extruded nucleus while some residual lectin is observed in the cytoplasm of circulating reticulocytes and erythrocytes.

Animals↗

Copper-binding proteins in liver of bluegills exposed to increased soluble copper under field and laboratory conditions.

Livers from bluegills exposed to increased soluble copper (Cu) under field and laboratory conditions were analyzed to determine the concentration and distribution of Cu in metalloproteins of different molecular size. Analyses were performed on bluegills collected from the impoundment of the H. B. Robinson Steam Electric Plant (Florence, SC) near the effluent discharge from the power plant, near the water intake to the cooling system, and from a control pond as well as on bluegills exposed under controlled laboratory conditions. Metalloproteins were separated into low molecular weight (LMW), intermediate molecular weight (IMW), and high molecular weight (HMW) fractions by using high-performance liquid chromatography. In the field-exposed bluegills, Cu concentrations in the LMW, IMW, and HMW fractions were highest in bluegills from the discharge site and lowest in those from the control pond. In the laboratory-exposed bluegills, Cu concentrations in the fractions increased with exposure concentration and time. Concentrations of Cu in the LMW protein fraction and pellet of bluegills exposed to 160 micrograms Cu/L appeared to plateau with long exposure times, whereas those in the HMW fraction continued to increase. Bluegills maintained in 80 micrograms Cu/L water at pH 5.5 accumulated lower concentrations of Cu in the LMW and pellet fractions and higher amounts in the HMW than in those maintained in 80 micrograms Cu/L at pH 7.0. Mortality was dependent on exposure concentration and duration and was higher in bluegills maintained in water at pH 5.5 than at pH 7.0.

Animals↗

Surface localization of an endogenous lectin in rabbit bone marrow.

A lectin, which may be involved in cell to cell adhesion during erythropoiesis in rabbit bone marrow, has been isolated and characterized. Several electron microscopical techniques have been used to investigate the cell surface distribution of this lectin in bone marrow utilizing colloidal gold conjugates of anti-lectin IgG or protein A. The lectin is present at the surface of erythroid cells at all stages of development but no lectin was detected on the surface of myeloid cells. The limitations and complementary nature of the techniques used are discussed.

Animals↗

Selective association of an endogenous lectin with connective tissues.

Using indirect immunofluorescence we have localized an endogenous beta-galactoside-specific lectin in resin-embedded rabbit tissue sections. The pattern of lectin distribution correlates well with biochemical estimations of lectin levels, being abundant in intestine, lung and heart tissue and relatively less abundant in skeletal muscle, liver and kidney. In all tissues lectin is found in connective tissue associated with fibroblasts and the extracellular matrix, and at the periphery of morphologically recognizable smooth muscle cells. The lectin is abundant in skin, intestine and blood vessels, where connective tissue forms the tissue architecture. It is also abundant in heart, where it is particularly associated with the capillaries and lung, where it is also found in alveolar cells. Discrete localization of lectin occurs in areas of connective tissue where epithelial elements are differentiating, such as the crypts of Lieberkuhns in the small intestine and hair follicles in the skin. From these observations we suggest that in cells of mesenchymal origin these endogenous lectins may play a role in the elaboration or organization of the extracellular matrix that regulates tissue differentiation in a number of embryonic and adult tissues.

Animals↗

Isoforms of an endogenous lectin in rabbit bone marrow.

A lectin which may mediate inter-erythroblast associations during red blood cell development in rabbit bone marrow has previously been purified and characterised. We have now detected different forms of this lectin in purified preparations and crude tissue extracts, by isoelectric focusing in agarose gels followed by rocket immunoelectrophoresis and by indirect antibody staining of focused proteins blotted onto nitrocellulose paper. These minor antigens are probably isoforms of the bone marrow lectin previously characterised.

Animals↗

Endogenous beta-galactoside-specific lectins in rabbit tissues.

Soluble beta-galactoside-specific lectins have been detected in many rabbits tissues and are relatively abundant in certain organs. These lectins are apparently identical to the lectin in rabbit bone marrow, which may be involved in inter-erythroblast associations during differentiation, and are very similar in structure and activity to lectins described in a number of species both in adult and embryonic tissues. The rabbit lectin is a monomer with a relative molecular mass of approximately 12 000 and has a low specific activity in the haemagglutination of trypsinized rabbit erythrocytes relative to lectins from other species, which are predominantly dimeric. In addition to the major antigen two minor isoforms of the rabbit galaptin were detected in all tissues. Since these lectins are widely distributed, it seems probable that they are involved in cellular functions that are prominent during development and also maintained in certain differentiated tissues. Experimental evidence currently available suggests that these lectins may be involved in the organization of extracellular matrix components.

Animals↗

An in vivo sister-chromatid exchange assay in the larvae of the mussel Mytilus edulis: response to 3 mutagens.

An in vivo sister-chromatid exchange (SCE) assay using the larvae of the mussel Mytilus edulis was developed. Larvae were exposed to mutagens beginning 12 h after fertilization and harvested 12 h later in the late trochophore stage. Dose responses to the mutagens bromodeoxyuridine, mitomycin C and methyl methanesulfonate were linear. The sensitivity of the SCE response of mussel larvae to these mutagens compares favorably with that of other systems and indicates this assay to be valuable in marine genetic toxicological monitoring.

Animals↗