The loci of action of ultraviolet and x-radiation and of photorecovery in the egg and sperm of the sea urchin Arbacia punctulata

Various experiments are described which show the locus of action of ultraviolet radiation and of x-ray in delaying cell division of the fertilized egg of Arbacia punctulata to be in the nucleus. Photorecovery following ultraviolet radiation has its locus in the egg cytoplasm. Analogy is drawn with the action of ultraviolet radiation and photorecovery on the E. coli-bacteriophage system, and certain suggestions are made regarding a common mechanism. Photorecovery after x-ray exposure could not be demonstrated.     

Ultrastructure of the unbalanced growth forms of Salmonellae obtained by the action of various factors

The unbalanced growth forms (UGF), also known as heteromorphous or pleomorphous forms, of salmonellae were obtained under the action of penicillin, homologous antiserum in combination with complement and lysozyme, as well as ultraviolet radiation. The effect of these factors was studied after their momentaneous action and in successive passages. The viability of UGF was determined in terms of colony-forming units. The appearance of UGF was induced by penicillin, a complex of immune substances and ultraviolet radiation, yet the action of two latter factors irreparably damaged most of the cells. The formation of UGF, showing stability in prolonged passages and capable of transformation into L-forms, occurred only under the influence of penicillin. Elementary bodies appeared in UGF, both capable and incapable of transformation into L-forms after successive passages. In the viable part of the UGF population obtained under the action of all the above factors damages in the peptidoglycan layer of the cell wall and, partly, in the cytoplasmic membrane, as well as disturbances in the process of cell division, linked with these cell elements, were observed.     

A comparison of five effects of ultraviolet light on the Arbacia egg

The demonstration of five different effects of ultraviolet radiation on the sea urchin's egg indicates that more than one basic photochemical process goes on there. Photorecovery is observed in only one of these. The need for caution in interpreting such effects is obvious. Evidence for a different mechanism for the timing of cleavage in eggs activated by ultraviolet radiation as compared to normally fertilized eggs is presented. The bearing of these studies on survival curves for microorganisms is discussed.     

Effect of ethylene on the endogenous cytokinin and gibberellin levels in tuberizing potatoes

High concentrations of 2-chloroethylphosphonic acid inhibited tuberization on aged potato tubers (Solanum tuberosum) that had been predisposed to the little tuber disorder. As a result of this treatment sprouts developed which contained relatively high levels of endogenous gibberellins and which elongated normally. The endogenous cytokinin levels in the different treatments did not change appreciably. It is suggested that tuberization is prevented by ethylene either as a direct inhibition of cell division or that it prevents the endogenous cytokinins from functioning. Irrespective of the mode of action of ethylene, cell division apparently is the primary process affected, the result being that storage tissue required for the accumulation of starch is not formed.     

Role of sulA and sulB in filamentation by lon mutants of Escherichia coli K-12.

Cells containing the pleiotropic Escherichia coli mutation lon filament extensively and die after exposure to ultraviolet light. Outside suppressors of the ultraviolet sensitivity, called sul, have previously been described at two loci; these mutations reverse the ultraviolet sensitivity of lon strains but do not affect the mucoidal or degradation defect of these strains. An isogenic set of strains carrying combinations of lon, sulA, and sulI was constructed, and their behavior during normal growth and after ultraviolet treatment was studied. sulA mutations had no detectable phenotype in lon+ cells; the lon sulA strains filamented transiently after ultraviolet irradiation, as did lon+ sul+ cells. We found that the sulB mutation, which alters cell morphology and slows recovery from transient filamentation after ultraviolet treatment, was epistatic to both lon and sulA. Whereas sulA mutations were recessive to the wild-type allele, sulB was partially dominant. The simplest model to account for our observations is that sulA and lon participate in a pathway of filamentation independent of that which produces transient filamentation in wild-type strains; sulB product may be the target of sulA action and may play a role in normal cell division.     

Inactivation of Neurospora conidia by ultraviolet radiation

The inactivation of Neurospora conidia by ultraviolet radiation is a consequence of the inactivation of the nuclei. The nuclei exhibit a kinetics of inactivation which is first order with respect to the number of (viable) nuclei. Arguments based on the efficiency of inactivation indicate that only quanta absorbed within a nucleus can lead to its inactivation.There are at least two sets of inactivation processes. One involves the production of lethal mutations, the other is a non-genetic (not inheritable) process. The two processes are apparently intimately connected: they yield the same action spectrum and the effects of each are reduced by photo reactivation. The assumption that the two processes are independent leads to predictions which are at variance with the observed dependence of the frequency of lethal mutations on dose of ultraviolet radiation.     

THE ACTION SPECTRUM OF SENSITIZATION TO HEAT WITH ULTRAVIOLET LIGHT

1. Heat does not sensitize paramecia to ultraviolet light but ultraviolet light sensitizes them to heat. Paramecia of two species (Paramecium caudatum and P. multimicronucleate) are much more readily killed by heat at 42.3° C. if they are first exposed to ultraviolet light. 2. From studies on paramecia irradiated with a given dosage at various wave lengths before being killed by heat, an action spectrum of the compound in the protoplasm being sensitized to heat can be determined. Proteins with absorption similar to that of pseudoglobulin are suggested by these experiments. 3. The effect upon living things differs from that on pure protein systems in that paramecia are not rendered more sensitive to temperatures below the lethal temperature whereas proteins are. 4. Almost complete recovery from ultraviolet light as judged by heat sensitivity occurs within 4 to 5 days. 5. By a study of the rate of recovery from doses at different wave lengths evidence suggesting effects on nucleic acid is obtained. 6. The possible significance of the data and the action spectrum is discussed.     

Chloroplast Growth and Replication in Germinating Spinach Cotyledons following Massive gamma-Irradiation of the Seed

Spinach seeds (Spinacia oleracea L.) given massive doses of γ-irradiation (500 krad) germinate and form a seedling with two green cotyledons and a radicle, but develop no further. Irradiated cotyledons show no increase in cell number or total DNA over a 7-day period in the light, while in control cotyledons there is a small increase in cell number and large increases in total DNA and chloroplast number. The chloroplasts of irradiated cotyledons are delayed in their division, become greatly enlarged and contain large amounts of starch. The whole population of chloroplasts subsequently undergoes a wave of division. The daughter chloroplasts show normal thylakoid development, but have some abnormal structural features caused by the radiation stress. Information on the effect of X-irradiation, ultraviolet irradiation, and 5-fluorodeoxyuridine on chloroplast replication and on chloroplast and nuclear DNA synthesis was obtained from cultured spinach leaf discs. It appears that chloroplast replication is more resistant to ionizing radiation than cell division and can proceed in the absence of nuclear DNA synthesis and greatly reduced chloroplast DNA synthesis.     

Socioeconomic status and non-melanoma skin cancer: A nationwide cohort study of incidence and survival in Denmark

Background: The two main types of non-melanoma skin cancer differ with the pattern of exposure to ultraviolet radiation (UVR): basal cell carcinoma (BCC) appears to be more closely related to intermittent solar exposure and sunburn, while the risk for squamous cell carcinoma (SCC) is a result of lifetime cumulated exposure to UVR. As these exposures may differ by social position, we investigated its role in the risk for and survival after BCC and SCC diagnosed in Denmark in 1994–2006 with follow-up through 2006. Methods: The analyses were based on 52,166 cases of BCC and 5033 cases of SCC in a cohort of 3.7 million people born between 1925 and 1976 and residing in Denmark in 1992–2006. Information on cancer cases and socioeconomic indicators were obtained from population-based registries. We used log-linear Poisson regression models to estimate incidence rate ratios and cumulative relative survival to estimate survival up to 10 years after the first incident cases of BCC and SCC. Results: High socioeconomic status, measured by both education and disposable income, was strongly associated with a higher risk for BCC, whereas there was no association between SCC and educational level and only a weak association with income. In general, relative survival after BCC was better than after SCC; the pattern of survival was not affected by socioeconomic indicators. Conclusions: The observed pattern of social status and risk for non-melanoma skin cancer differed substantially for the two cancer types, supporting the hypothesis that they may have different aetiologies.     

PHOTOSYNTHETIC INSENSITIVITY OF THE TERRESTRIAL CYANOBACTERIUM NOSTOC FLAGELLIFORME TO SOLAR UV RADIATION WHILE REHYDRATED OR DESICCATED1

Photosynthetic performance of the terrestrial cyanobacterium Nostoc flagelliforme (M. J. Berkeley et M. A. Curtis) Bornet et Flahault during rehydration and desiccation has been previously characterized, but little is known about the effects of solar UV radiation (280–400 nm) on this species. We investigated the photochemical activity during rehydration and subsequent desiccation while exposing the filamentous colonies to different solar radiation treatments. Photochemical activity could be reactivated by rehydration under full‐spectrum solar radiation, the species being insensitive to both ultraviolet‐A radiation (UVAR; 315–400 nm) and ultraviolet‐B radiation (UVBR). When the rehydrated colonies were exposed for desiccation, the effective PSII photochemical yield was inhibited by visible radiation (PAR) at the initial stage of water loss, then increased with further decrease in water content, and reached its highest value at the water content of 10%–30%. However, no significant difference was observed among the radiation treatments except for the moment when they were desiccated to critical water content of about 2%–3%. At such a critical water content, significant reduction by UVBR of the effective quantum yield was observed in the colonies that were previously rehydrated under indoor light [without ultraviolet radiation (UVR)], but not in those reactivated under scattered or direct solar radiation (with UVR), indicating that preexposure to UVR during rehydration led to higher resistance to UVR during desiccation. The photosynthetic CO₂ uptake by the desiccated colonies was enhanced by elevation of CO₂ but was not affected by both UVAR and UVBR. It increased with enhanced desiccation to reach the maximal values at water content of 40%–50%. The UV‐absorbing compounds and the colony sheath were suggested to play an important role in screening harmful UVR.     

UV-B radiation and selenium affected energy availability in green alga <Emphasis Type="Italic">Zygnema</Emphasis>

Green alga Zygnema was exposed to three concentrations of selenium and two levels of UV-B radiation. The combined effects of both treatments on energy availability; photochemical quantum yield and respiratory potential were studied. Our findings show that traces of selenium enhance metabolic process connected with photochemical quantum yield and mitochondrial respiration. Surprisingly, selenium does not diminish the effects of UV-B radiation; on the contrary, the combined action of UV-B radiation and traces of selenium leads to pronounced negative effects on photochemical quantum yield and the respiratory potential. Selenium is involved in the activation of energy resources in green alga Zygnema. The importance of selenium for activity of the mitochondria is possibly an evolutionary recollection from an endosymbiotic bacterium.     

Genetic recombination in Coprinus: III. Inlluence of gamma-irradiation and temperature treatments on meiotic recombination

Non-lethal doses of gamma-irradiation (5 krad) increased meiotic recombination in Coprinus lagopus when treatments were given at the beginning of karyogamy. The division stage at this time was judged to be late leptotene and the duration of the sensitive period was assessed to be 3–4 h. In C. lagopus the radiation-sensitive stage is distinct from the cold-sensitive stage (pachytene). The additive effect of irradiation at early karyogamy followed by cold treatment in pachytene suggested that the two factors influenced different steps in the recombination process. On the other hand, irradiation followed by heat treatment did not significantly alter recombination frequency as compared to single treatments. It was surmised that radiation and high temperature act on the same factor(s) or at the same steps to bring about a similar net result. It was suggested that irradiation at leptotene may cause single-strand breaks in DNA which eventually participate in exchange.     

STUDIES ON CELL METABOLISM AND CELL DIVISION : IV. COMBINED ACTION OF SUBSTITUTED PHENOLS,CYANIDE, CARBON MONOXIDE,AND OTHER RESPIRATORY INHIBITORS ON RESPIRATION AND CELL DIVISION

The effects of 4,6-dinitro-o-cresol and 2,4,5-trichlorophenol on the respiration and cell division of fertilized eggs of Arbacia punctulata have been determined in the presence of each of a number of respiratory inhibitors. The experimental results obtained appear to afford some understanding of the mechanism of action of the substituted phenols on respiration and on cell division. 1. From the fact that the stimulated respiration is completely cyanide and carbon monoxide sensitive, it may be concluded that all of the extra oxygen uptake induced in Arbacia eggs by 4,6-dinitro-o-cresol passes through the metal containing oxidase system. All of the extra oxygen uptake also passes through oxidative steps which can be poisoned by non-stimulating phenols like 2,4-dinitrothymol and 4-nitrocarvacrol, by phenylurethane, by 5-isoamyl-5-ethyl barbituric acid, by malonic acid, or by iodoacetic acid. To abolish all respiratory stimulation by suboptimum concentrations of 4,6-dinitro-o-cresol, each of these inhibitors must be present in a concentration which reduces the normal respiration in the absence of substituted phenols by at least 20–40 per cent. 2. The degree of reduction of the stimulated respiration by a given concentration of carbon monoxide or potassium cyanide depends on the concentration of 4,6-dinitro-o-cresol or 2,4,5-trichlorophenol, being most marked in suboptimum concentrations and least marked in greater than optimum concentrations of the substituted phenol. In contrast to this result, the reduction of the stimulated respiration by a given concentration of 5-isoamyl-5-ethyl barbituric acid or malonic acid is least marked in suboptimum concentrations and most marked in greater than optimum concentrations of the substituted phenol. 3. The present experiments appear to indicate that the inhibition of cell division by substituted phenols is not attributable to a direct action of these agents on mitotic processes nor to an overstimulation of any respiratory process. The inhibition of cell division appears to be associated with the inhibition, by the substituted phenols, of some component of the cyanide sensitive respiratory system. This inhibition is of such a type as to allow the overall respiration to proceed at a rate in excess of the control value, even when division is completely suppressed. The dependence of the division mechanism on a respiratory step which is relatively hypersensitive to poisoning by the substituted phenols is comparable to the dependence of the Pasteur reaction in certain normal and tumor tissues on an oxidative step which is specifically poisoned by the substituted phenols (16). The substituted phenols have no inhibiting effect in vitro on the principal metal containing respiratory catalysts or the principal dehydrogenases; they also do not inhibit the fermentative reactions involved in the anaerobic glycolysis of fertilized Arbacia eggs. It is therefore suggested that the respiratory inhibiting and division inhibiting effects of the substituted phenols may be attributable to the action of these substances on one or more of the oxidation-reduction or phosphorylating steps which are involved in the transfer of hydrogen from the dehydrogenase systems to the specifically cyanide sensitive oxidase mechanism of the eggs. The identification of the respiratory step poisoned by the substituted phenol would constitute an interesting contribution to the chemistry of cell division and experiments to this end are now in progress.     

Photobiophysics of furanocoumarins

Furocoumarins (psoralens) are photosensitizers of plant origin, which increase the sensitivity of biological objects to near ultraviolet (UV-A, 320-400 nm). In combination with UV-A, they are successfully used for treating many dermal and autoimmune diseases (PUVA therapy and photophoresis). Along with therapeutic effects, the furocoumarin photochemotherapy induces a number of side-effects (erythema, edema, hyperpigmentation, and premature aging of skin). All photobiological effects of furocoumarins result from their photochemical reactions. Therefore, in order to advance the therapy, it is necessary to know the photochemical mechanisms of induction of both side- and therapeutic effects. The types of photoreactions of furocoumarins classified with respect to reactive photoproducts interacting with substrate were considered. Primary emphasis was placed on reactions proceeding with the participation of photooxidation products of furocoumarins. Among these photoproducts, at least two types can be distinguished. Some of them possess membranotoxic properties, others produce the immunosuppressory action in vivo. The photochemical mechanisms of the formation of the photoproducts of furocoumarins are different. It was found that, by varying the illumination conditions (intensity of UV-A radiation or the concentration of the photosensitizer), it is possible to obtain the photoproducts of furocoumarins that have either membranotoxic or immunosuppressory properties. It was found that the mechanisms of the immunosuppressive action of the photooxidation products of furocoumarins have some features in common with those underlying the PUVA therapy and photophoresis. It is assumed that the photochemical basis of the therapeutic action of furocoumarins is the reactions with the involvement of the products of their photooxidation.     

Photoinactivation of Ichthyrotoxin from Axenic Cultures of Prymnesium parvum Carter

The ichthyotoxin of Prymnesium parvum is inactivated by visible (400 to 510 mμ) as well as by ultraviolet light (255 mμ). The changes in the absorption spectrum of purified (pigment-free) ichthyotoxin during this process indicate that different mechanisms may be involved in the inactivation by visible or ultraviolet light. Photoinactivation is not affected by the presence of cells, cell pigments, oxygen, or glutathione.     

LEAF DEVELOPMENT OF RUMEX PATIENTIA L. (POLYGONACEAE) EXPOSED TO UV IRRADIATION (280-320 NM)

Two factors which affect leaf ontogeny and ultimate leaf size: (1) the rate and duration of cell expansion, and (2) the rate and duration of cell division, were examined for their role in the slower early leaf growth rate and the smaller size of fully expanded leaves of plants exposed to ultraviolet-it (UV-B 280-320 nm) radiation. Rumex patientia L. was grown in controlled environment chambers under enhanced UV-B radiation (equivalent to daily solar UV-B irradiation at 40°N latitude in mid-May with an atmospheric ozone concentration of 0.20 atm-cm) and control treatments. The pattern of growth as expressed in changes of mean cell size in two distinct cell types, tissue cell density, and length of the entire leaf blades are consistent with the hypothesis that the radiation primarily affects cell division rather than cell expansion. Furthermore, it appears that the radiation probably alters the rate rather than the duration of the cell division phase. An understanding of the mechanism of radiation damage should facilitate prediction of how this stress may interact with other stresses to which plants are normally subjected. Species with normally prolonged periods of cell division during leaf expansion may be particularly impacted if solar UV radiation were intensified as a result of atmospheric ozone reduction.     

Pattern of unequal cell division and development in Caulobacter crescentus

The pattern of asymmetric division has been examined in Caulobacter crescentus (gram-negative aquatic bacteria) by determining the position of the “division site” on cells of different ages. Measurements of cell width and length at this site, which corresponds to the point of eventual cell separation, were made on electron micrographs of cells stained with phosphotungstic acid. The results show that (i) the division site is formed early in the cell cycle and it constitutes the first visible feature on the growing stalked cell to differentiate the incipient swarmer cell, (ii) the division site is located asymmetrically (closer to the swarmer pole than the stalked pole) on the dividing cell, (iii) its position relative to the stalked and swarmer poles does not change during the cell cycle, and (iv) division is consequently unequal, with the swarmer cell always smaller than the stalked cell. The implications of these findings for general models of unequal cell division and stem cell development are discussed.     

Videomicroscopic extraction of specific information on cell proliferation and migration in vitro

In vitro cell imaging is a useful exploratory tool for cell behavior monitoring with a wide range of applications in cell biology and pharmacology. Combined with appropriate image analysis techniques, this approach has been shown to provide useful information on the detection and dynamic analysis of cell events. In this context, numerous efforts have been focused on cell migration analysis. In contrast, the cell division process has been the subject of fewer investigations. The present work focuses on this latter aspect and shows that, in complement to cell migration data, interesting information related to cell division can be extracted from phase-contrast time-lapse image series, in particular cell division duration, which is not provided by standard cell assays using endpoint analyses. We illustrate our approach by analyzing the effects induced by two sigma-1 receptor ligands (haloperidol and 4-IBP) on the behavior of two glioma cell lines using two in vitro cell models, i.e., the low-density individual cell model and the high-density scratch wound model. This illustration also shows that the data provided by our approach are suggestive as to the mechanism of action of compounds, and are thus capable of informing the appropriate selection of further time-consuming and more expensive biological evaluations required to elucidate a mechanism.     

Action of colcemid in sea urchin eggs

The effects of Colcemid, the deacetyl-N-methyl derivative of colchicine, on the eggs of Arbacia punctulata were investigated. Colcemid in concentrations of 2.7 x 10^-5 M or greater blocks syngamy (the fusion of the pronuclei) in these eggs. Although a tenfold decrease in concentration of Colcemid usually permits the pronuclei to fuse, the subsequent division is blocked. In the sea urchin egg, the duration of presyngamy is about 15 min during which time there is no DNA synthesis. However, DNA synthesis is recorded in Colcemid-blocked cells prior to syngamy. Radioautographs of Colcemid-blocked cells which were immersed into thymidine-³H exhibited silver grains above each of the pronuclei. The action of Colcemid on Arbacia eggs is reversible. Nevertheless, exposures to 2.7 x 10^-5 M Colcemid for only 3 min, initiated 5 min after insemination, caused delays of 70 min in subsequent division. In general, cells are more sensitive to Colcemid prior to the time when the mitotic spindle is being assembled than at presyngamy stages. The results are discussed in terms of Colcemid action on pronuclear fusion and cell division.