THE ACTION SPECTRUM OF DAPHNIA MAGNA (CRUSTACEA) PHOTOTAXIS IN A SIMULATED NATURAL ENVIRONMENT

Abstract— The action spectrum of phototaxis in Daphnia magna (Crustacea) was measured in a chamber which simulated a natural angular distribution of underwater light. A 17% step-down in irradiance was used to stimulate the phototactic response at all wavelengths and irradiances tested. Peaks in the spectral response curves depended on the fluence rate to which the zooplankton were acclimated. The wavelength of maximum response (Z_max) shifted from yellow-green at the highest acclimation fluence rate (5.1 × 10⁻² Wm⁻²) to blue-violet at moderate rates. At low acclimation fluence rates, the blue-violet maximum was retained and another maximum developed in the red. At the lowest fluence rate (1.6 × 10⁻⁵ Wm⁻²), the blue-violet and red maxima were lost and another maximum developed in the near ultraviolet. The action spectrum indicates the presence of three, and possibly four, photopigments with Z_max, at ∼405, 440, 570 and 690nm. The 440 and 690nm maxima may belong to the same photopigment; however, this was not tested. Changes in zooplankton swimming speed, caused either by large changes in irradiance or by mechanical stimuli, were accompanied by changes in the strength of the phototactic response to the −17% stimulus at any irradiance level for white and monochromatic light, and indicated the presence of a mechanism connecting swimming speed and photosensitivity.     

EFFECTS OF INTENSITY AND FLUENCE UPON DNA SINGLE-STRAND BREAKS INDUCED BY EXCIMER LASER RADIATION

Abstract— A Xenon-chloride excimer laser emitting energy at 308 nm was used to induce single-strand breaks (SSBs, frank breaks plus alkali-labile lesions as assayed by alkaline sucrose sedimentation techniques) in purified DNA from Bacillus subtilis . A fluence response study and a peak pulse intensity study were performed. At a pulse energy of 0.1 mJ/pulse, the radiation induced SSBs in a linear fashion (91 SSB/10⁸ Da per MJ/m²) to a maximum exprimental fluence of 1.28 MJ/m². The pulse intensity study showed that there were no significant changes in DNA breakage (105 SSB/10⁸ Da) between 2.93 times 10⁹ and 5.86 times 10¹¹ W/m² (0.11 and 22.0 mJ/pulse) at a constant total fluence of 1.1 MJ/m² (27000 mJ dose). This study has verified and extended previous work by quantifying the yield of SSBs induced in DNA by this laser radiation.     

ANALYSIS OF MULTIPLE PHOTORECEPTOR PIGMENTS FOR PHOTOTROPISM IN A MUTANT OF Arabidopsis thaliana

Abstract
The shape of the fluence-response relationship for the phototropic response of the JK224 strain of Arabidopsis thaliana depends on the fluence rate and wavelength of the actinic light. At low fluence rate (0.1 μmol m^-2s^-1), the response to 450-nm light is characterized by a single maximum at about 9 μmol m^-2. At higher fluence rate (0.4 μmol m^-2s^-1), the response shows two maxima, at 4.5 and 9 μmol m^-2. The response to 510-nm light shows a single maximum at 4.5 μmol m^-2. Unilateral preirradiation with high fluence rate (25 μmol m^-2s^-1) 510-nm light eliminates the maximum at 4.5 μmol m^-2 in the fluence response curve to a subsequent unilateral 450-nm irradiation, while the second maximum at 9 μmol m^-2 is unaffected. Based on these results, it is concluded that a single photoreceptor pigment has been altered in the JK224 strain of Arabidopsis thaliana.     

EFFECT OF INTRACELLULAR INJECTION OF EGTA AND TETRAETHYLAMMONIUM CHLORIDE ON THE RECEPTOR POTENTIAL OF LOCUST PHOTORECEPTORS

Abstract— The depolarizing photoreceptor potential of locust, Valanga irregularis , as that of other arthropods, has an initial transient phase that declines to a stable sustained phase. In this work, the effect of intracellular injection of EGTA and TEA on these phases was studied.
(1) Reversal potential of the transient phase was higher than that of the sustained phase.
(2) EGTA and TEA enhanced both phases and the enhancement was more prominent in the sustained phase.
(3) Resting potential was shifted slightly upward by the injection. The amplitude of the maximum response was also increased.
(4) Current injected through an electrode filled with 2 M KCl had a significant effect on the shape of the response depending on the direction of the current.
(5) It is suggested that the activation of K⁺ conductance induced by an increase of intracellular free Ca²⁺ concentration causes a decrease in amplitude of the sustained phase.     

Nonmonotonous variation of DNA angular separation during asymmetric pulsed field electrophoresis

Asymmetric pulsed field electrophoresis within crystalline arrays is used to generate angular separation of DNA molecules. Four regimes of the frequency response are observed, a low frequency rise in angular separation, a plateau, a subsequent decline, and a second plateau at higher frequencies. It is shown that the frequency response for different sized DNA is governed by the relation between pulse time and the reorientation time of DNA molecules. The decline in angular separation at higher frequencies has not previously been analyzed. Real‐time videos of single DNA molecules migrating under high frequency‐pulsed electric field show the molecules no longer follow the head to tail switching, ratchet mechanism seen at lower frequencies. Once the pulse period is shorter than the reorientation time, the migration mechanism changes significantly. The molecule reptates along the average direction of the two electric fields, which reduces the angular separation. A freely jointed chain model of DNA is developed where the porous structure is represented with a hexagonal array of obstacles. The model qualitatively predicts the variation of DNA angular separation with respect to frequency.     

Molecular reorientation dynamics and microscopic friction in liquids

Molecular rotation reorientation times are investigated using time resolved fluorescence depolarization studies of three solutes of similar size and shape (nile red, neutral nile blue and cationic nile blue) dissolved in alcohol and alkane solvents as well as an extensive compilation of previous results for neautral and charged solutes dissolved in non-polar, polar and associated solvents. A universal correlation is foung between reorientation time, solvent viscosity, and solute volume for solutes dissolved in alkanes, while strongly interacting solutes experience relatively enhanced friction, and non-polar solutes dissolved in alcohols experience reduced friction. The results are compared and contrasted with slip and stick hydrodynamic predictions, and used to develop empirical correlations, which can be used to predict molecular reorientation times with an uncetainty on the order of a factor of two in virtually any solute-solvent system.     

Stochastic simulation of laminar and electroosmotic convection in capillary electrophoresis and electrochromatography.

A stochastic (Monte Carlo) simulation has been utilized to study the combination of laminar and electroosmotic convection under typical conditions for capillary electrophoresis and electrochromatography. The radial velocity profiles are determined for varying proportions of laminar and electroosmotic flow at a constant linear velocity. In addition, the molecular zone profiles and the corresponding statistical moments are established for nonretained and retained solutes. It is found that the radial velocity profile has significant influence for solutes that are not retained by a stationary phase, including those separated by electrophoresis alone. However, this effect is rapidly diminished such that retained solutes with an absorption coefficient as small as 0.1 show little influence if their diffusion coefficients in the mobile phase are on the order of 1.0 x 10(-5) cm2/s. Larger solutes with diffusion coefficients of 1.0 x 10(-6) cm2/s or less show slightly greater variance and asymmetry. In most cases, however, slow kinetics and resistance to mass transfer in the stationary phase exceed that in the mobile phase. As a consequence, there is little effect of the radial velocity profile for retained solutes.     

Stepwise Two-Photon Excitation of 1,5-Dihydroflavin Mononucleotide: Study of Flavosemiquinone Properties

Abstract— Photoionization of 1,5-dihydroflavin mononucleotide in aqueous solution, leading to formation of hydrated electrons and flavosemiquinone radicals, was studied by nanosecond laser photolysis (excitation at LD = 354.7 nm). Analysis of the laser fluence dependence of the photoionization yield indicates that the reaction is a stepwise two-photon reaction with the fluorescent state of the flavin as intermediate step. Extinction coefficients of the neutral and anionic flavosemiquinone were obtained from transient absorbance measurements at 310-660 nm. The pro-tolytic dissociation constant of the neutral radical was determined from a study at pH 7.3-10.3 of the spectral evolution in the submicrosecond time range due to de-protonation of this radical. The value pK_a= 8.7 0.1 was obtained. The formation of tautomeric species of the neutral flavosemiquinone was suggested based on the existence of a spectral evolution of this species during the first microseconds after excitation.     

DOUBLET Stentor DO NOT DISPLAY PHOTODISPERSAL

Normal Stentor, called singlets since they have a single membranellar band and oral groove surrounding their frontal field, swim away from light sources and collect in the darker areas of an unevenly illuminated container (photodispersal). Phenotypic variants, called doublets since they have 2 membranellar bands and 2 oral grooves, do not exhibit this behavior. Doublets produce photophobic responses and contractions when illuminated at the same fluence rates which produce those responses in singlets, hence their sensitivity to light is normal. Illumination of the frontal field of doublets produces a photophobic response at lower fluence rates than does illumination of their side or posterior. This directional sensitivity is quantitatively similar to that observed in singlets. However, doublets do not reorient their swimming direction after a phobic response as extensively as do singlets. This failure in reorientation is the probable reason that doublets fail to show photodispersal. These results imply that the mechanism producing photodispersal in singlets depends on photophobic responses or some other, presently undescribed, response which requires an asymmetric frontal field.     

THE EFFECT OF CYTOKININS ON GROWTH and PIGMENT ACCUMULATION OF RADISH SEEDLINGS (RAPHANUS SATIVUS L.) GROWN IN THE DARK and AT DIFFERENT LIGHT QUANTA FLUENCE RATES*

Abstract— The dependency of cytokinin effects upon irradiance was studied with radish seedlings ( Raphanus sativus L. cv. Saxa Treib). Kinetin (6-furfurylamino-purine) or BAP (6-benzylamino-purine) were applied via the roots of plants growing either in continuous darkness or under high (90 Wm^-2) or low intensity white light (10Wm^-2). Apart from the different development of plants at low and high fluence rates, the following cytokinin effects were found:
(1) Both cytokinins acted in a similar manner on growth characteristics and pigment accumulation at high and low light conditions, BAP being in many cases more effective than kinetin.
(2) When compared with the control, the cytokinins suppressed hypocotyl and root lengthening in the dark and light-grown plants. In darkness they led to increased cotyledon areas, whereas in the light the leaf expansion was suppressed.
(3) In the etiolated and low light grown plants, the anthocyanin content of the hypocotyls was enhanced due to the action of cytokinins, whereas under high light the anthocyanin accumulation was decreased.
(4) In the cotyledons of etiolated plants, more phototransformable protochlorophyll(ide) and more carotenoids were formed when cytokinins were present. In green leaves the carotenoid content was diminished due to the action of cytokinins, particularly in plants grown in strong light. The chlorophyll a/b ratio was increased in the cytokinin-treated plants in most cases.
The results suggest a light dependency of the cytokinin effects. It is believed that the response of a plant towards exogenously applied cytokinins is similar to that with high intensity light.     

FLUENCE-RATE DEPENDENCE OF MONOPHOTONIC REACTIONS OF NUCLEIC ACIDS IN VITRO AND IN VIVO

The Bunsen-Roscoe law, also known as the reciprocity law ( E = f(F) with F = I t ) has only limited validity for monophotonic reactions of nucleic acids. Especially at low fluence rates, the extent of in vitro and in vivo photoreactions of nucleic acids in the far-UV and near-UV range is a function of the fluence and of the fluence rate ( E = f (F;I)). In vitro experiments with poly(dA)poly(dT) clearly show that the far-UV (254 nm) response, indicated by the changes of the ellipticity at 315 nm, does not obey the Bunsen-Roscoe law at low fluence rates in the range between 1 W m^-2 and 20 W m^-2. In vivo experiments with Escherichia coli revealed very similar anomalies. Studying the growth delay after irradiation with far-UV light at 280 nm or near-UV light at 334 nm, we have confirmed the lack of reciprocity in both spectral ranges. The failure of the Bunsen-Roscoe law for the 280 nm and 334 nm UV irradiation effect at low fluence rates was in the range O < I < 40 W m^-2. In both cases reciprocity occurred at higher fluence rates (40 < I < 100 W m^-2).     

Na+/K+ TRANSPORT AND PHOTOSENSITIVITY OF THE COLORLESS FLAGELLATE Peranema trichophorum (Euglenida)

Abstract— Perimycin, ouabain and elevation of extracellular K⁺ concentrations cause an increase in the fluence rate thresholds (white light) for the step-up photophobic response in Peranema trichophorum . Elevation of extracellular Na⁺ concentration decreases the thresholds for this response in comparison to the control level. The fluence rate threshold of perimycin-treated cells increases before the side effect of an antibiotic action appears. Removal of K⁺ ions from the medium of K⁺-treated cells to a concentration of 1 mM depresses the threshold for the step-up response to the control level. By addition of K⁺ or Na ⁺ ions to perimycin- or ouabain-treated cells the threshold returns to the control value. It is suggested that the flagellar and cell membrane are responsible for changes of P. trichophorum photosensitivity.     

THE ROLE OF HEMOGLOBIN IN THE PHOTOTAXIS OF THE NEMATODE Mermis nigrescens

Abstract A dense zone of crystalline hemoglobin in the head has been presumed to be involved in the photosensitivity of Mermis. With the aim of identifying its role, we have studied the wavelength dependence of the phototaxis. Measuring phototaxic efficiency at constant photon fluence rate (intensity), we find that the spectral response curve is approximately fiat from 350–540 nm and falls to an insignificant level by 580 nm. This is unlike the absorptance (fraction absorbed) spectrum of the hemoglobin pigmentation. Also, fluence-rate/response curves at 420 and 500 nm occur at the same fluence rates even though these wavelengths correspond to a maximum and a minimum of hemoglobin absorption. These results prove that the hemoglobin cannot be functioning as the visual pigment in phototaxis but, for reasons discussed, they neither confirm nor rule out a role as a shadowing pigment. The results are consistent with a shadowing role in the presence of contrast enhancement by the nervous system.
A steep fluence-rate dependence suggests that contrast enhancement does occur in Mermis phototaxis. The 420 or 500 nm fluence rate for half-maximal response is 6 times 10 photons s^-1 cm^-2 (about equivalent in effectiveness to pre-dawn twilight). The wide range of sensitivity, 350–560 nm, has interesting implications as to the nature of the visual pigment.
†NATO reaearch collaborator a n leave from the Department of Biophysics. Laboratorium voor Algemene Natuurkunde, Rijksuniversiteit Groningen, Westersin-gel 34. 9718 CM Groningen, The Netherlands.     

THE PHOTOMOVEMENT OF Caenorhabditis elegans, A NEMATODE WHICH LACKS OCELLI. PROOF THAT THE RESPONSE IS TO LIGHT NOT RADIANT HEATING

Abstract— Caenorhabditis elegans adults were tested at constant temperature with 10 s periods of monochromatic light alternated with 20 s dark periods. Stimuli at effective intensities and wavelengths caused an increase in the frequency of ecclitic (phobic, avoidance) responses, which was measured as an increase in the probability of a temporary reversal in direction of movement. For monochromatic stimuli ranging from 420 to 680 nm at a constant 56 picoeinsteins s^-1 cm^-2, only those at520–600 nm elicited significant responses. At 540 nm the threshold fluence rate was approximately 30 pE s^-1 cm^-2. At saturating intensities the mean reversal probability was increased to 0.20 in 10 s from a background level of 0.12. approximately.
Because C. elegans lacks ocelli and is very sensitive to temperature, possible sources of radiant heating were considered in detail, including (a) infrared present in the stimuli, (b) absorption of light by the arena, and (c) absorption of light by a nematode pigment. All possible sources were found to cause a negligible temperature rise, on the order of or less than the natural temperature fluctuations inside the worm, 1.5 times 10^-6°C. A 2 times 10^-4°C temperature rise produced by a 1230 nm infrared stimulus had no significant effect on reversal frequency. It was concluded that the response to illumination must have been to light, and not to temperature changes.
Large, + or - 2 °C changes from the acclimation temperature caused significant increases in the background frequency of ecclitic responses (a thermoecclisis or thermoklinokinesis). However, neither the threshold nor the saturation level of light-induced responses was affected by the ± 2°C changes.     

Host sensitization of Tb3+ ions in tribarium lanthanide borates Ba3Ln (BO3)3 (Ln = Lu and Gd)

The vacuum-ultraviolet (VUV) spectroscopic properties of undoped and Tb(3+)-doped borates Ba(3)Ln(BO(3))(3) (Ln = Lu and Gd) with different crystal structures were investigated by using synchrotron radiation. Ba(3)Lu(BO(3))(3) (BLB) crystallizes in a hexagonal structure, whereas Ba(3)Gd(BO(3))(3) (BGB) crystallizes in a trigonal structure. The maximum host absorption for BLB and BGB was found to locate at ~179 and ~195 nm, respectively. Upon host excitation, BLB exhibits an intrinsic broad UV emission centered at 339 nm, which is attributed to the recombination of self-trapped excitons that may presumably be associated with band-gap excitations or molecular transitions within the BO(3)(3-) group. In contrast to BLB, no broad emission but line emission ascribed to a Gd(3+)(6)P(J)-(8)S(7/2) transition was observed in the emission spectrum of BGB. Upon doping of Tb(3+) ions into the hosts of BLB and BGB, an efficient energy transfer from the host excitations to Tb(3+) via host/Gd(3+) emission was observed, showing that host sensitization of Tb(3+) occurs in these rare-earth borates.     

CALCIUM CONTROL OF PHOTOTACTIC ORIENTATION IN Chlamydomonas reinhardtii: SIGN AND STRENGTH OF RESPONSE

Abstract— Chlamydomonas reinhardtii responds to a blue light stimulus by an oriented swimming (phototaxis) toward or away from the stimulus source. In this study it is established that the sign and strength of the phototactic response are a complex function of extracellular [Ca²⁺], stimulus fluence rate, time of analysis after onset of stimulation and light pretreatment. At very low extracellular [Ca²⁺] the response is weak and usually negative. At [Ca²⁺] close to the preconditioning level, phototactic response becomes stronger and positive. As [Ca²⁺] is raised further, the initial (2 s) response remains positive but the long term (20 s) becomes negative and very strong. At extremely high [Ca²⁺] the cells become immobile. This bimodal behavior suggests that two different mechanisms determine the direction of the turn. Data cannot be explained in terms of a simple model. The model which accounts for most of the details of the behavior is that of Kamiya and Witman (1984), which proposes that positive response is triggered by a transient increase in intracellular [Ca²⁺] and negative response by a decrease below unstimulated level of Ca²⁺, at least in the range of 10^-9-10^-6 M [Ca²⁺]. The strong negative orientation which follows an initial positive response above this level of [Ca²⁺], in these experiments, is best explained by an adaptation of the cells due to an increased (on average) intracellular [Ca²⁺].     

PHEOMELANIN PHOTOCHEMISTRY. PHOTOLYSIS OF MODEL COMPOUNDS

Abstract— Phytochrome control of nitrate reductase activity has been studied in cotyledons and hypocotyls of light-grown Sinapis alba. Under polychromatic irradiation, an increase in the fluence rate of far-red light added to a constant source of photosynthetically active radiation causes a decrease in the phytochrome photoequilibrium and, in the hypocotyl, this results in decreased nitrate reductase activity. However, in the cotyledons this difference is only observed transiently. In both organs, enzyme activity is correlated with the level of the far-red light absorbing form of phytochrome, P_fr. These correlations are not altered when the fluence rate (with respect to phytochrome) is increased, suggesting that the responses are not fluence rate dependent. The results obtained are consistent with the notion that in fully de-etiolated seedlings, P_ft alone controls nitrate reductase activity.     

Suppression of Delayed-type Hypersensitivity and Hemolysis Induced by Previously Photooxidized Psoralen: Effect of Fluence Rate and Psoralen Concentration

Abstract— The kinetics of the formation of biologically active psoralen photooxidation (POP) products were analyzed by the biological effects produced. Effects of the UV light fluence rate and psoralen concentration during the preir-radiation were investigated to assess the yield of POP products, which were active in vivo (inducing suppression of delayed-type hypersensitivity [DTH] reaction to sheep red blood cells) and in vitro (altering the human erythrocyte membrane permeability). It was shown that the reciprocity law of the irradiation fluence rate and time was not valid in the case of POP-induced hemolysis and DTH suppression. Immunosuppressive POP products were more efficiently formed at low fluence rate (20.8 W/m²), whereas POP hemolysins were more efficiently produced at a high fluence rate (180 W/m²) of UV light. The yield of immunosuppressive POP products was enhanced in dilute psoralen solutions, while the POP hemolysins yield increased with increasing psoralen concentration. A kinetic scheme for psoralen photoproduct formation was proposed. Kinetic analysis showed that a labile intermediate was produced as the result of excitation of psoralen. This intermediate was either converted to a stable immunosuppressive POP product, or two intermediates combined to form a POP hemolysin. It is proposed that PUVA therapy conditions are more favorable for the formation of immunosuppressive rather than membrane-damaging psoralen photooxidation products.     

BLUE LIGHT INDUCED PHOTOTRANSFORMATION OF PHYTOCHROME IN THE PRESENCE OF FLAVIN*

Abstract— Irradiation of the P_r form of phytochrome in the presence of flavin mononucleotide (FMN) which absorbs the actinic blue light yields P_fr at a rate greater than that in the absence of FMN. The actinic blue light absorbed by FMN enhances the phototransformation of P_r via the energy transfer from the former to the latter. On the other hand, the photoreversion of P_fr was inhibited by the presence of FMN when illuminated with blue light. The lack of photo-enhancement of the reversion of P_r, by blue light suggests that the P_fr chromophore (acceptor) transition dipole is virtually perpendicular to the FMN transition dipole, as the result of a chromophore reorientation in the P_r→P_fr phototransformation. The fact that blue light absorbed by flavin preferentially enhances the forward phototransformation of phytochrome while inhibiting the reversion may have an important implication in the high irradiance responses in plants in terms of a preferential accumulation of P_fr by blue light excitation.