PHOTOPHOBIC BEHAVIORAL RESPONSES OF EUGLENA IN A LIGHT INTENSITY GRADIENT AND THE KINETICS OF PHOTORECEPTOR PIGMENT INTERCONVERSIONS*

Abstract— Accumulation of Euglena gracilis in small illuminated regions called light traps is due to a phobic response to the diminished light intensity at the boundary of the region. The rate of such accumulation of cells was measured as functions of both the light intensity within the trap and the change of intensity at the boundary of the trap. The initial rate of accumulation of a population of cells was taken to be a direct measure of the phobic response of a single typical cell. The data indicate that the strength of the behavioral response in a single cell may be described as being proportional, to the rate of change of the amount of photochemically active form of a photoreceptor pigment molecule which can exist in two predominant forms.     

PHOTOSENSORY TRANSDUCTION IN CILIATES. II. POSSIBLE ROLE OF G-PROTEIN AND cGMP IN Stentor coeruleus

Abstract— The heterotrichous ciliate, Stentor coerulus , exhibits a welll defines photophobic response to a sudden increase in the intensity of visible light. the phobic reactions usually appear with a latency perios (i.e. a time delay between the onset of the stimulus and the stop response). This latency of phobic response was significatly increased when the cells werw incubated with 8-bromo-guanosine3',5'-cyclic monophospjhate. In the presence of this nucleotide, a reduction of cell responsiveness (i.e. the number of photophobically responding cells) was also observed. similar effects were observed when cells were treated with pertussis toxin, a G-protein activity modulator, and 3'-isobutyl-methylxanthine, an inhibitor of guanosine 3', 5'-cyclic monophosphate (cGMP) phosphodiesterase. the G-protein activator fluoroaluminate and 6-anilino-5,8-quinolinedione (LY 83583) (an effective agent for lowerin cellular cGMP levels) showed opposite effects on hte cell photophobic response. These result indirectly suggesnt that the level of cytoplamic cGMP, possibly modulated by a G-protein-coupled CGMP phosphodiesterase, plays a phototreasducing role in Stentor . In addition, using an antiserum raised against bovine transducin, a cross reacting protein with an apparent molecular mass of 39 kDa was detected on immunoblots. The α-subunits of a Stentor G-protein has also been partially cloned and sequenced. However, the possible coupling between the G-protein and the putative phosphodiesterase remains to be established.     

ACTION SPECTRUM AND ELECTROPHYSIOLOGICAL RESPONSES CORRELATED WITH THE PHOTOPHOBIC RESPONSE OF STENTOR COERULEUS

Abstract— The blue-green ciliate. Stentor coeruleus , is found predominantly in shady places. This concentration occurs because stentor responds when swimming from a shaded area to a lighted area by reversing the direction of its ciliary beat and reorienting its swimming direction until it once again is in the shaded area. A graded receptor potential is recorded from microelectrodes in vacuoles of stentor when the animal is photically stimulated. For all but very weak stimuli this receptor potential is sufficient to elicit a regenerative transmembrane response of variable amplitude in a swimming animal. Suprathreshold electrical stimuli also elicit this regenerative response. In turn the regenerative response is coupled to ciliary reversal. Thus ciliary reversal appears to be produced whenever the photic receptor potential crosses the threshold for elicitation of the regenerative response.
Using the threshold for production of ciliary reversal as a criterion response, an action spectrum was obtained. This action spectrum correlates well with the absorption spectrum of the major pigment of S. coeruleus , stentorin. Stentor bleached of pigment also have an elevated threshold for ciliary reversal. Thus stentorin seems to be the photosensitive pigment in stentor responsible for its photophobic behavior.     

PHOTOSENSORY TRANSDUCTION IN CILIATES. I. AN ANALYSIS OF LIGHT-INDUCED ELECTRICAL AND MOTILE RESPONSES IN Stentor coeruleus

Abstract— Light-induced membrane potantial changes and motile responses have been studied in Stentor cells with intracellular microelectrodes and video microscopy, respectively. Intracellulae microelectrode showed that step-up increase in light induced an electrical membrane response which consisted of an initial membrane depolarization (photoreceptor potential) followed by an action potential and maintaining phase of depolarization (afterdepolarization). The amplitude of the receptor potetial is dependent on the intensity of light stimulus and the action potetials appears with a lag period (latency) after the onset of light stimuklus. The extent of the membrane established between the latency for te action poitential and the onset of ciliary reversal (stop responses). A time correlation was also observed between the duration of the membrane afterdepolarization and the duration of backward swimming. the action spectrum for the photoreceptor potential amplitude of Stentor resembled the action spectra for the latency of ciliary reversal and the photoresponsiveness, iondicating that the photomovement response and membrane potential changes are coupled through the same photosensor system. A hypothesis on the photosensory transduction chain in Stentor is discussed according to ehich the photoreceptors and the ciliary apparatus is mediated by the membrane potential canges.     

THE EFFECT OF POTASSIUM IODIDE ON PHOTOPHOBIC RESPONSES IN EUGLENA: EVIDENCE FOR TWO PHOTORECEPTOR PIGMENTS*

Abstract— Potassium iodide, a quencher of flavin fluorescence, inhibits the shock reaction which Euglena experiences upon a sudden decrease in light intensity (inverse photophobic response) completely at a concentration of 150 mM. The rate of swimming of the cells at the same concentration of KI is reduced to 30% of the control. The direct photophobic response, a shock reaction which appears identical but occurs upon an increase in light intensity, is unaffected by KI as is negative phototaxis of Euglena. It is concluded that a non-flavin pigment system mediates photoreception for the direct photophobic response and negative phototaxis.     

PHOTOSENSORY TRANSDUCTION IN CILIATES. III. THE TEMPORAL RELATION BETWEEN MEMBRANE POTENTIALS AND PHOTOMOTILE RESPONSES IN Blepharisma japonic urn

Abstract— Blepharisma japonicum exhibits a step-up photophobic response when subjected to an increase in light stimulus intensity. This response is characterized by the stop reaction after a period of delay followed by backward swimming (lateral rotation). The latency of the stop response decreased and duration of the lateral rotation increased as the intensity of light stimuli was raised. A step-increase in light intensity elicited a graded membrane depolarization (photic receptor potential), as measured by intracellular microelectrode. When the amplitude of receptor potential exceeded a threshold depolarization for membrane excitation (15–25 mV), an all-or-none action potential of 50–65 mV in amplitude was evoked which also occurred with some latency. Light stimuli of higher intensity (suprathreshold) elicited action potential which was followed by a membrane after-depolarization. Increasing the intensity of stimuli caused generation of an action potential with shorter lag period and prolonged after-depolarization. The action spectra for the latency of stop reaction, receptor potential amplitude and cell photoresponsiveness showed maxima at 460, 530 and 580 nm. The analysis of temporal relationships between the electrophysiological responses and the motile events showed that latency of an action potential, induced by the receptor potential, correlates well with the latency of a cell stop response. Also the duration of membrane after-depolarization resembled the time period of the cell's backward swimming (cell rotation). The data obtained indicate that the primary reaction initiated by light absorption in the photoreceptor pigment (blepharismin) is converted into the observed electrical potential changes, which in turn results in the photomotile response of Blepharisma cells.     

THE PHOTOPHOBIC RESPONSE OF VARIOUS SULFUR AND NONSULFUR PURPLE BACTERIA

Abstract— The photophobic response of 34 strains of nonsulfur and sulfur purple bacteria was examined with respect to response-eliciting light intensities. The bacteria were grown in defined synthetic media or in Winogradsky columns. Two population methods based on Engelmann's light trap were used to determine the discrimination thresholds of the bacteria. A single-side irradiation method allowed the estimation of approximate values, while the double-side irradiation method provided more exact values of the discrimination threshold. Sixteen strains belonging to 9 different species exhibited discrimination thresholds between 0.7% and 2.6%. The motility of the other 18 strains proved to be insufficient to measure light sensitivities with the methods used. The effect of various environmental factors on the light sensitivies of Chromatium vinosum D and Rhodospirillum rubrum Ha was examined. The measurements and observations made in this work recommend strains of Rhodospirillum rubrum and Chromatium vinosum as model organisms for further studies.     

THE ROLE OF CALCIUM IN Chlamydomonas PHOTOMOVEMENT RESPONSES AS ANALYSED BY CALCIUM CHANNEL INHIBITORS

Phototaxis and light-induced stop responses in Chlamydomonas are known to be calcium dependent. We show that phototaxis is stereoselectively inhibited by dihyropyridines, verapamil, diltiazem, omega-conotoxin and pimozide, all inhibitors of slow L-type calcium channels. In contrast, the stop response in Chlamydomonas can be specifically reduced only by omega-conotoxin and pimozide. The light-regulated calcium uptake as detected by 45calcium can be completely suppressed by verapamil and omega-conotoxin but not by diltiazem or any of the dihyropyridine-type calcium channel inhibitors. We conclude that phototaxis and stop response in Chlamydomonas are regulated by three distinguishable drug receptor sites. One of them controls phototaxis and is sensitive to verapamil. The second site controls stop response and phototaxis and shows a high sensitivity to omega-conotoxin and pimozide. These two drug receptors seem to be localized in the plasma membrane and function as ion channels. In addition, calcium influences internal signal transduction from the photoreceptor to the flagella. This internal role of calcium is inhibited by the dihydropyridine binding to a dihydropyridine receptor protein. The arylazide-1,4-dihydropyridine[3H]azidopine binds with a Kd = 35 nM to a 50 kDa protein located in one of the internal cell membranes. Azidopine binding is fully reversible and can be partially inhibited by nimodipine and PN-200110. This protein is the first identified dihyropyridine receptor in an unicellular plant cell. It might serve as an internal calcium regulating channel in Chlamydomonas.     

FLUORESCENCE INDUCTION IN ALGAE AND CHLOROPLASTS

Abstract —The first phases of the fluorescence transient elicited by illumination of dark-adapted algae or isolated chloroplasts (biphasic rise φ_***v - φ_I - _P) have previously been shown to be controlled by two quenchers: Q , the primary acceptor of Photosystem 2 interacting with the secondary acceptor pool A ; and R , a non-photochemical quencher which goes into a non-quenching state as A is reduced.
The dependence of the kinetics of φ decay after illumination upon the redox state of A was studied. It is suggested that some of the centres are in a disconnected state, where electron transfer between Q and A is hindered, the amount of such centres being correlated to the reduction state of A . The implications of this hypothesis on the problem of the variation of the Q-A 'equilibrium constant' under different experimental conditions, and on Murata's 'weak light effect' are discussed.
The effect of 3-(3,4 dichlorophenyl)-1,1 dimethylurea (DCMU) on R is shown to depend upon the redox state of A . A DCMU-induced shift of the midpoint potential of R may account for this dependence.
Evidence is given suggesting that the transient reduction of A which occurs in algae during the φ_I-φ_P rise is controlled by an induction process on the acceptor side of Photosystem 1.     

ACTION SPECTRA OF THE PHOTOPHOBIC RESPONSE OF BLUE AND RED FORMS OF Blepharisma japonicum

Abstract— When exposed, in the presence of molecular oxygen, to light intensities of the order of3–30 W m^-2, the ciliate Blepharisma japonicum changes its color from red to blue, because of the photooxidation of the photoreceptor pigment, blepharismin, to pxyblepharismin. Both red-and blue-pigmnentes cells show step-up photophobic responses. The action spectra f the light-dependent behaviour of the red and the blue form of Blepharisma have been determined; their structure is very similar to that the photosensing and phototransducing properties of blepharismin are maintained in its photooxidized form. oxyblepharismin.     

LIGHT-INDUCED SHIFTS IN THE ABSORPTION SPECTRUM OF CAROTENOIDS IN RED AND BROWN ALGAE

Abstract— Changes in light absorption in the region 450–540 nm were observed upon illumination of the red algae Iridaea splendens, Schizymenia pacifica and Porphyra perforata and the brown alga Pheostrophion irregulare . The difference spectra of these changes in Iridaea and Schizymenia showed maxima and minima at about 465, 480, 495, 515 nm. The spectra were similar to difference spectra earlier observed in photosynthetic bacteria, and the location of the maxima and minima suggested a shift towards longer wavelength of a compound with absorption maxima at about 440, 470, and 500 nm, probably a carotenoid. Similar, but more distorted difference spectra were observed in the other algae. Time courses and size of the signals induced by light of different wavelengths suggest that excitation of both photosynthetic pigment systems causes a shift in carotenoid absorption, with kinetics which appear to be similar to those of the well-known change at 515 nm in green plants.     

铜及其配合物在生物体系中的作用

自人类有历史记载以来，铜及其配合物即用作药物。铜是人体必需的金属元素。人体的正常代谢过程需要它们，但无法体内合成，因此，需要每天从饮食摄取和吸收。生物药学家和研究者以极大的努力彻底地了解铜及其配合物在生物体系中的作用，希望获得更多地信息以利于人类疾病的预防和治疗。本综述评述了铜在组织中的分配和代谢、铜依赖酶、在病态时铜的非正常代谢、铜配合物的药疗活性和抑制氧自由基的产生。     

PHOTOREGULATION OF NITRATE UTILIZATION IN GREEN ALGAE AND HIGHER PLANTS*

Abstract— Nitrate reductase from eukaryotes can be reversibly inactivated, blue light being an effective activating agent both in vitro and in vivo. Hydroxylamine proved to be a powerful inactivating agent of Ankistrodesmus braunii nitrate reductase. Irradiation with blue light of NH₂OH-inactivated nitrate reductase, specially in the presence of μM amounts of FAD, promoted the recovery of the enzyme activity. Similarly, photoexcited methylene blue reactivated spinach nitrate reductase. On the other hand, in vitro nitrate reductase is highly susceptible to photodynamic inactivation caused by singlet O₂. Aerobic incubation of the active spinach enzyme with either FMN or methylene blue under either blue or red light respectively led to its irreversible inactivation. Irradiation of frozen and thawed spinach leaf discs also promoted, in situ, an irreversible inactivation of nitrate reductase, provided that 6₂ was present in the incubation mixture. Thus, either in vitro or in situ, light can cause two quite different responses of nitrate reductase, its blue light-dependent photoactivation in a flavin sensitized reaction and its photodynamic inactivation in a singlet O₂-dependent process.     

可逆与不可逆氢在甲烷化反应中的作用

用自行设计和建立的加压动态分析装置研究了甲烷化催化剂上氢的吸附和反应行为．结果表明，在反应条件下吸附的氢可区分为不可逆吸附氢和可逆吸附氢两类，不可逆吸附氢又可分为能被ＣＯ顶替出来的和不能被顶替的两种．起甲烷化反应的是可逆吸附氢，而能被ＣＯ顶替出来的不可逆氢对吸附ＣＯ起促进作用，不能被ＣＯ顶替出来的不可逆氢是甲烷化催化剂必不可少的组分（或称“促进剂”）．     

PASSIVE AND ACTIVE ROLE OF FLUORESCERS IN THE CHEMILUMINESCENCE OF TETRAMETHOXYDIOXETANE

Abstract— Tests for the possible catalytic role of fluorescers in the chemiluminescent decomposition of dioxetanes are discussed and applied to the case of tetramethoxydioxetane. This dioxetane, which can undergo direct thermal decomposition yielding excited methyl carbonate with subsequent excitation of fluorescers (for example 9, 10-dibromoanthracene. DBA), is shown to be sensitive also to catalysis by fluorescers such as rubrene. 9, 10-diphenyl-anthracene (DPA) and 9, IOdicyanoanthracene (DCNA). The possibility of a charge-transfer mechanism of chemiluminescent catalysis is suggested.     

THE ROLE OF FRONTAL NEURONS IN PAIN AND ACUPUNCTURE ANALGESIA

To study the role of frontal neurons in pain and acupuncture analgesia, experiments were carried out on 4 monkeys (Macaca mulatta) during performing two tasks, the Heat Discrimination Response GO/NO-GO Task (Task Ⅰ) and the Heat Delayed Discrimination GO/NO-GO Task (Task Ⅱ). After a criterion of 90% of correct performances over three successive days had been reached, activity of single neurons was recorded from the frontal cortex. Of 276 Task Ⅰ-related neurons, 211 responded to noxious and/or innocuous heat stimuli. Of 73 Task Ⅱ-related neurons, 59 responded to noxious and/or innocuous heat stimuli. During acupuncture, the nociceptive reaction time for lever release to avoid painful stimuli was increased, the percentage of correct performances was lowered, and the neuronal responses to noxious and/or innocuous stimuli were suppressed. These neurons were mainly located in a circumscribed area medial to the superior ramus of the arcuate sulcus in frontal cortex, including the prefrontal and premotor a     

THE ROLE OF SUPEROXIDE AND SINGLET OXYGEN IN LIPID PEROXIDATION

Abstract— An investigation into the mechanism of lipid peroxidation catalyzed by xanthine oxidase showed a dependence upon superoxide, singlet oxygen and adenosine 5'-diphosphate chelated iron (ADP-Fe³⁺). In the absence of ADP-Fe³⁺ or in the presence of superoxide dismutase there is complete inhibition of enzymatic peroxidation. Initiation of peroxidation likely occurs through an ADP-perferryl ion complex formed by ADP-Fe³⁺ and superoxide. Use of the singlet oxygen trapping agent 2,5-diphenylfuran showed that singlet oxygen does not participate in the initiation of peroxidation but rather in the propagation of peroxidation. The mechanisms of NADPH-cytochrome P450 reductase-catalyzed and ADP-Fe²⁺ catalyzed lipid peroxidation parallel that of xanthine oxidase in that initiation occurs through a superoxide dismutase-sensitive reaction and that singlet oxygen is present during propagation of lipid peroxidation.     

可逆与不可逆吸附CO在合成甲醇反应中的作用

合成甲醇反应是CO+H_2反应系列中较简单的反应之一,它与同系列的甲烷化反应有着不同的反应条件.从热力学观点来看,甲烷化反应和合成甲醇反应同是体积缩小反应,加压应该有利于两反应的进行,实际上合成甲醇反应需加压,而甲烷化反应却不一定需要加压.最近我们用动态分析技术对烃的临氢转化、乙炔加氢和苯加氢反应研究发现,在实际反应条件下,反应物在催化剂表面上的吸附可分为可逆与不可逆吸附两类,它们在多相催化反应中对活性、选择性和稳定性均有影响,但是它们对操作压力的依赖程度是不一样的.因此研究