Lethal and sub-lethal effects of UVB on juvenile Biomphalaria glabrata (Mollusca: Pulmonata)

Although Schistosoma mansoni occurs mainly in the tropics, where intense levels of solar radiation are present, the impact of ultraviolet (UV) light on schistosome transmission is not known. The purpose of this study was to investigate potential effects of UVB (290-320nm) on juvenile Biomphalaria glabrata, the snail intermediate host of S. mansoni. Albino and wild-type snails were exposed to doses of UVB from UV-fluorescent lamps, and the following were measured: survival, photoreactivation (light-mediated DNA repair), effects on feeding behavior, and morphological tissue abnormalities. Irradiation with UVB is lethal to B. glabrata in a dose-dependent manner. Exposure to white light subsequent to UVB irradiation enhances survival, probably by photoreactivation. The shell offers some, but not complete, protection. Experiments in which UVB transmittance through the shell was blocked with black nail polish suggest that injury to both exposed (headfoot) and shell-enclosed (mantle and visceral mass) tissues contributes to mortality in lethally irradiated snails. Wild-type (pigmented) snails are less susceptible to lethal effects of UVB than albino snails, and they may be more capable of photoreactivation. UVB exposure inhibits snail feeding behavior, and causes tentacle forks and growths on the headfoot. Thus, UVB may influence the life cycle of S. mansoni by both lethal and sub-lethal damage to the snail intermediate host. However, the ability of snails to photoreactivate may mitigate these effects.     

Excision repair in xeroderma pigmentosum group C but not group D is clustered in a small fraction of the total genome

DNA repair in xeroderma pigmentosum complementation groups C and D occurs at a low level. Measurements of pyrimidine dimers remaining in bulk DNA from the whole genome indicated very little excision in either complementation group. The repair sites in group C cells were, however, clustered together in small regions of the genome which appeared to be mended nearly as efficiently as the whole genome is mended in normal cells, while repair in group D cells was randomly distributed. Growth of normal cells in cycloheximide or 3-aminobenzamide neither inhibited repair nor altered the distribution of repair sites. Growth of normal cells in novobiocin or aphidicolin inhibited excision but repair remained randomly distributed. On the basis of these observations, and consideration of other cellular features of group C and D, we suggest that group C may represent a mutation which results in a low level of repair enzymes with normal function. Group D, on the other hand, may represent a mutation resulting in functionally defective repair enzymes.     

Repair-deficient xeroderma pigmentosum cells made UV light resistant by fusion with X-ray-inactivated Chinese hamster cells.

Xeroderma pigmentosum (XP) is an autosomal recessive human disease, characterized by an extreme sensitivity to sunlight, caused by the inability of cells to repair UV light-induced damage to DNA. Cell fusion was used to transfer fragments of Chinese hamster ovary (CHO) chromosomes into XP cells. The hybrid cells exhibited UV resistance and DNA repair characteristics comparable to those expressed by CHO cells, and their DNA had greater homology with CHO DNA than did the DNA from XP cells. Control experiments consisted of fusion of irradiated and unirradiated XP cells and repeated exposure of unfused XP cells to UV doses used for hybrid selection. These treatments did not result in an increase in UV resistance, repair capability, or homology with CHO DNA. The hybrid cell lines do not, therefore, appear to be XP revertants. The establishment of these stable hybrid cell lines is an initial step toward identifying and cloning CHO DNA repair genes that complement the XP defect in human cells. The method should also be applicable to cloning genes for other diseases, such as ataxia-telangiectasia and Fanconi's anemia.     

DISTRIBUTION OF DIATOMS IN THE PLANKTON OF YAQUINA ESTUARY,OREGON1

The distributional patterns of diatoms in the plankton of the Yaqnina Estuary, Oregon, were, investigated and related to selected climatic and hydrographic factors. Distribution was strongly influenced by seasonal patterns of rainfall resulting in the introduction of a large volume of fresh water into the estuary during fall and winter. Plankton assemblages in spring, summer and fall had fewer diatom species and exhibited a more rapid rate of change in species composition than in winter. Winter assemblages were further characterized by many pennate diatoms, apparently dislodged from the benthos during periods of high freshwater discharge and silt loads. A statistical measure of community difference indicated an increase in taxonomic homogeneity among assemblages throughout the estuary with the onset of the rainy season in late fall and a gradual transition to a more heterogeneous system again during late spring. Canonical correlation ordered 20 prominent diatom taxa along the salinity gradient and identified possible relationships among certain taxa and selected environmental variables, namely visible light energy and temperature. Redundancy in the species data given the environmental data was only 40%, emphasizing the difficulty in demonstrating a quantitive relationship between plankton dynamics in the field and concurrent measurements of chemical and physical variables.     

Sublethal effects of ultraviolet b radiation on miracidia and sporocysts of Schistosoma mansoni: intramolluscan development, infectivity, and photoreactivation

Schistosoma mansoni occurs in tropical regions where levels of ultraviolet B (UVB; 290-320 nm) light are elevated. However, the effects of UVB on parasite transmission are unknown. This study examines effects of UVB on the miracidia and sporocysts of S. mansoni, focusing specifically on intramolluscan development, infectivity, and the ability to photoreactivate (repair DNA damage using visible light). Histology revealed that miracidia irradiated with 861 J x m(-2) underwent abnormal development after penetrating Biomphalaria glabrata snails. Total number of sporocysts in snail tissues decreased as a function of time postinfection (PI), among both nonirradiated and irradiated parasites; however, this decrease was greater in the latter. Moreover, whereas the proportion alive of nonirradiated sporocysts increased PI, that of irradiated sporocysts, i.e., derived from irradiated miracidia, decreased. Irradiation of miracidia with UVB resulted in decreased prevalence of patent infection (defined by presence of daughter sporocysts) in a dose-dependent manner, and no infections occurred at a dose of 861 J x m(-2). Like many aquatic organisms, including the snail host, parasites were able to photoreactivate if exposed to visible light following UVB irradiation, even subsequent to penetrating snails. These photoreactivation results suggest cyclobutane-pyrimidine dimers in DNA as the primary mechanism of UVB damage, and implicate photoreactivation, rather than nucleotide excision, as the main repair process in S. mansoni.     

CELL SURVIVAL CHARACTERISTICS AND MOLECULAR RESPONSES OF ANTARCTIC PHYTOPLANKTON TO ULTRAVIOLET-B RADIATION1

Twelve species of Antarctic diatoms were studied to assess UV sensitivity in relation to cellular and molecular aspects of DNA damage and repair. Responses of cell survival, induction of DNA damage, and DNA repair capacity were determined. There was a wide range of interspecific UV-sensitivity among diatoms. D₃₇ values (average fluence to kill one cell) ranged from 681 J · m^?2 (most sensitive) to 25,338 J · m^?2 (most resistant). Molecular analysis (by radioimmunoassay) of UV-induced DNA damage [induction of cys-syn cyclobutane dimers and pyrimidine (6-4) pyrimidone photoproducts] also revealed considerable variability among species [0.98–84 lesions · (10⁸ daltons DNA)^?1 induced by exposure to 2500 J · m^?2]. Repair of DNA damage ranged from 0.18 to 2.72 lesions removed · (10⁸ daltons DNA)^?1 in 6 h; removal represented 0.72–73.5% of initial damage. Comparison of cellular responses associated with photoenhanced repair and nucleotide excision (“dark”) repair indicated that light-mediated correction of UV damage was an important factor in cell survival. There was a relationship between the number of photoproducts induced and cell survival, but not between repair efficiency and survival. The data also indicate a general dependence of photoproduct induction and D₃₇ values on cell size and shape (expressed as the surface area: volume ratio which ranged from 0.07 to 0.66 between species) and suggest that these factors are indicators of UV sensitivity. Smaller cells with greater surface area: volume ratios sustained more damage per unit of DNA, had lower D₃₇ values, and were more sensitive to UV exposure. The wide species variations observed in molecular and cellular responses to UV exposure emphasize the ecological implications of changes in natural UV regimes. These changes can act as determinants of cell size and taxonomic structure within phytoplankton communities and have as yet unknown effects on trophic interactions within the Antarctic ecosystem.     

Distribution of mycosporine-like amino acids in the sea hare Aplysia dactylomela: effect of diet on amounts and types sequestered over time in tissues and spawn

We investigated the interaction of diet and accumulation of UV-absorbing mycosporine-like amino acids (MAAs) in body tissues and spawn of the sea hare Aplysia dactylomela to determine if MAA accumulation reflects type and level of dietary intake. Food sources were the red algae Acanthophora spicifera, Centroceras clavulatum, and Laurencia sp., and the green alga, Ulva lactuca. Adults were maintained on these foods for 40 days, after which feces were collected and tissues separated by dissection. Field animals were similarly sampled at this time. All spawn from experimental and field animals was collected over the study period. Samples, including seaweed foods, were analysed for six MAAs. Overnight consumption experiments using a variety of common seaweeds and one seagrass from A. dactylomela's habitat showed that the four seaweeds selected as foods were among those best-eaten by Aplysia. After 40 days levels of specific MAAs in the tissues of experimental animals showed excellent correlation with those in their diets, suggesting that the MAAs were dietarily-derived. Relative MAA contents in spawn from all diet groups correlated well with those in spawn from field animals. Commonest MAAs in spawn were porphyra-334, shinorine, and palythine, in this order. Concentrations of these MAAs were maintained at constant levels over time in spawn from all diet groups eating red algae and from field animals. Spawn from the Ulva dietary group showed an initial significant decline in MAA concentrations, but levels stabilized after the first 2 weeks. Skin was rich in porphyra-334 and shinorine, and levels of these in experimental animals correlated well with comparable levels in the skin of field animals. Digestive glands contained high levels of asterina-330, particularly those of the Centroceras dietary group, where concentrations reached a maximum of 21 mg dry g(-1).     

Temporal patterns and variations in phytoplankton community organization and abundance in Narragansett Bay during 1959-1980

Incident irradiance, surface water temperature and phytoplanktonspecies abundances were measured at weekly intervals in NarragansettBay from 1959 through 1980. Stepwise discriminant analyses (SDA)of this 22-year data set indicate that fundamental ecosystemchanges occurred between the 1960s and 1970s, with 1969 beingthe key transitional year in these decadal shifts in phyto planktontaxonomic structure and seasonal abundance. This decadal shiftwas accompanied by the increased summer abundance of small Thalassiosiraspp., which first appeared in 1966 and by 1969 became the sixthmost important phytoplankton component in this bay. Decadaltrends in phyto plankton community organization and abundancewere also accompanied by distinct long-term climatological gradientsof temperature and light. The 1960s were generally colder andbrighter than the 1970s. Prior to 1969, the annual phytoplanktonmaximum occurred most commonly during winter; in the 1970s,the annual maximum generally shifted to a summer event. Three5-year phytoplankton cycles occurred between 1959 and 1974.During each pentade, the phytoplankton community returned toa similar taxonomic organization and abundance cycle after divergingin the intervening years. Pentade cycles did not occur after1974; the phytoplankton community thereafter diverged significantlyfrom each preceding year. Five species [Skeletonema costatum,Detonula confervacea, Asterionellopsis glacialis, Hererosigmaakashiwo (= Olisthodiscus lureus) and Thalassiosira nordenskioeldii]dominated the phytoplankton over the 22-year period. SDA revealeda high degree of similarity and constancy in the annual occurrencepatterns of these taxa. The decadal shifts revealed by SDA weremore directly related to the considerable interannual variabilitythat characterized the abundance and seasonality of the lessabundant species.     

Influence of Ozone-Related Increases in Ultraviolet Radiation on Antarctic Marine Organisms

Every spring for the past two decades, depletion of stratosphericozone has caused increases in ultraviolet B radiation (UVB,280–320 nm) reaching Antarctic terrestrial and aquatichabitats. Research efforts to evaluate the impact of this phenomenonhave focused on phytoplankton under the assumption that ecosystemeffects will most likely originate through reductions in primaryproductivity; however, phytoplankton do not represent the onlysignificant component in ecosystem response to elevated UVB.Antarctic bacterioplankton are adversely affected by UVB exposure;and invertebrates and fish, particularly early developmentalstages that reside in the plankton, are sensitive to UVB. Thereis little information available on UV responses of larger Antarcticmarine animals (e.g., birds, seals and whales). Understandingthe balance between direct biological damage and species-specificpotentials for UV tolerance (protection and recovery) relativeto trophic dynamics and biogeochemical cycling is a crucialfactor in evaluating the overall impact of ozone depletion.After more than a decade of research, much information has beengathered about UV-photobiology in Antarctica; however, a definitivequantitative assessment of the effect of ozone depletion onthe Antarctic ecosystem still eludes us. It is only obviousthat ozone depletion has not had a catastrophic effect in theAntarctic region. The long-term consequences of possible subtleshifts in species composition and trophic interactions are stilluncertain.     

DNA repair in man: regulation by a multigene family and association with human disease

The major mechanism of repair of damage to DNA involves a conceptually simple process of enzymatic excision and resynthesis of small regions of DNA. In man and other mammals, this process is regulated by several gene loci; up to 15 mutually complementary genes or gene products may be involved. Repair deficiency results in an array of clinical symptoms in skin, central nervous system, and hematopoietic and immune systems, the major example being xeroderma pigmentosum (XP), a disease with a high incidence of cancer. Cloning repair genes by straightforward methods has proved difficult, but we have begun the effort by demonstrating that correction of a human repair deficiency can be achieved by transferring very small fragments of DNA from normal hamsters into XP cells. One of the complementation groups of XP cells (group C) appears to express a change in gene regulation such that these cells repair only a small clustered region of the DNA with high efficiency.