Electrotonic Coupling between Adjacent Internodal Cells of Chara braunii: Transmission of Action Potentials beyond the Node

Many nodal cells are interposed between two internodal cellsof Chara braunii. When an action potential conducted in an internodereached the node, no electrical activation in the nodal cellscould be found, although an area of the membrane bordering thenodal cells in this internode was partially activated (end-membraneaction potential). When the action potential approached thenode along the stimulated internode, an electrotonic potentialchange (depolarization) was produced in the other internode.This depolarization was greatly depressed by the end-membraneaction potential of the stimulated internode, so that hardlyany transmission took place. The ratio of the potential changein the surface membrane of the adjoining ("postsynaptic") internode(cell b) to that of the stimulated one (cell a), the couplingratio, e_b/e_a, can be estimated from a simple equivalent circuitof the nodal region composed of two surface-membrane resistances(R_a, R_b) and intercellular resistance (R_n). If R_n remains thesame, a higher ratio should be produced with a larger R_b, butthe ratio does not depend on any change in R_a, which could beproved experimentally. Transmission of the action potentialbeyond the node was frequent when the coupling ratio was increasedand when the threshold that elicits the action potential waslowered by immersing the node in a K or Na salt solution. ¹ Present address: Department of Physiology, Tohoku UniversitySchool of Dentistry, Sendai 980, Japan. (Received December 1, 1980; Accepted January 23, 1981)     

Ephaptic Transmission and Conduction Velocity of an Action Potential in Chara Internodal Cells Placed in Parallel and in Contact with One Another

When two separated Chara internodal cells were kept in contactover a length of 14 mm or more in moist air, an action potentialof one cell could be transmitted to the other cell in about40% of cases (ephaptic transmission). The action potential ofthe former cell was sometimes eliminated transiently when anew action potential was elicited in the latter cell. The newaction potential reactivated the former cell. The conductionvelocity of the action potential was reduced from 0.30?0.11cm/s (mean?SD) to 0.15?0.05 cm/s by ephaptic transmission ofthe action potential. Substitution of an artificial pond waterfor moist air or contact over a shorter length (4 mm insteadof 14 mm or more) between the two cells reduced the couplingratio, the ratio of the change in membrane potential of onecell to that of the other cell, and no transmission was observed.After ephaptic transmission, the action potentials of the twocells were conducted at almost the same velocity along the cells.The velocity was increased to 0.68?0.06 cm/s by simultaneousconduction of the action potentials in the two adjacent cells.When the simultaneously conducted action potentials reachedthe node of one cell, at the point at which another cell madecontact, they were able very frequently to trigger a new actionpotential in the adjoining cell. (Received September 28, 1989; Accepted April 5, 1990)     

Inhibitory Effect of N,N'-Dicyclohexylcarbodiimide on Reduction of Cytochrome b560 and Stoichiometry of the Reduction in Chromatium Chromatophores

N,N'-Dicyclohexylcarbodiimide (DCCD) inhibited the flash-inducedreduction of cytochrome b₅₆₀, by blocking the electron flowbetween the secondary electron acceptor and cytochrome b₅₆₀presumably in the vicinity of the ubiquinone pool. The stoichiometryof the reduced cytochrome b₅₆₀ per reaction center bacteriochlorophylldimer was 0.77?0.12 throughout the redox potential range of150 to 390 mV at pH 7.0. The high stoichiometry suggested thatmost of the electrons ejected from the reaction center reducedcytochrome b₅₆₀. (Received January 19, 1982; Accepted March 15, 1982)     

Changes in the Levels of Chlorophyll and Light-Harvesting Chlorophyll a/b Protein of PS II in Rice Leaves Aged under Different Irradiances from Full Expansion through Senescence

Effects of irradiance on changes in the amounts of chlorophyll(Chl) and light-harvesting chlorophyll a/b protein of PS II(LHCII) were examined in senescing leaves of rice (Oryza sativaL.). Results of treatments at two irradiances (100% and 20%natural sunlight) were examined after the full expansion ofthe 13th leaf throughout the course of senescence. With 20%sunlight, the Chl content decreased only a little during leafsenescence, while with 100% sunlight it decreased appreciably.Similarly, the amount of LHCII protein during treatment with20% sunlight remained almost constant. However, the ratio ofChl a/b during the shade treatment decreased significantly andthe rate of decrease was greater than during the full-sunlighttreatment. The ratio of Chl a/b for Chl a and b bound to LHCIIwas about 1.2, irrespective of leaf age or irradiance treatment.When the amounts of Chl bound to LHCII were calculated fromthe total leaf content of Chl and the ratio of Chl a/b, assuminga ratio of Chl a/b bound to LHCII of 1.2, they were well correlatedwith the amounts of LHCII protein. Changes in the amounts of LHCII synthesized during the two irradiancetreatments were examined using an ¹⁵ tracer. Incorporation of¹⁵N into LHCII declined dramatically during both treatmentsfrom full expansion through senescence, suggesting that therewas little synthesis of LHCII protein during that time. In addition,the amount of LHCII synthesized during senescence was lowerduring the shade treatment than during the 100% sunlight treatment.These results indicate that the absence of an apparent changein levels of LHCII with shade treatment during senescence wascaused by the very low rate of turnover of LHCII protein. (Received June 17, 1992; Accepted September 28, 1992)     

The Influence of Stress Conditions on Chlorophyll Content of Two Range Grasses with Contrasting Photosynthetic Pathways

The influence of various treatments and temperature regimeson total chlorophylls and on the chlorophyll a:b ratio of westernwheatgrass and blue grama plants was investigated at differenttime intervals during the 120-day growth period. Western wheatgrass,a C₃ species, accumulated greater amounts of chlorophyll thandid blue grama plants, a C₄ species. Maximum concentrations(mg gd wt^–1) of chlorophylls in western wheatgrass andin blue grama were recorded at the lower (13/7°C) and higher(30/18°C) temperature regimes. Nitrogen fertilizer alonedecreased the chlorophyll content in both species. The chlorophylla:b ratio in blue grama ranged from an average of 2·00under irrigated plus fertilized conditions to 3·00 undercontrol and fertilized conditions. On the other hand, the chlorophylla:b ratio in western wheatgrass remained constant at 3·00throughout the growing season under various treatments and temperatureregimes.     

Propagation of Action Potential over the Trap-lobes of Aldrovanda vesiculosa

In the trap-lobes of Aldrovanda vesiculosa, an action potentialwas generated in a cell located at the base of a sensory hairstanding on the margin of central portion of the paired lobes,which spread over this portion within about 40 msec. The electricalcoupling ratio for two adjacent cells in the middle layer ofthe lobes was 0.8. This showed that an action potential generatedin a cell of the trap-lobes must spread electrotonically toadjacent cells. An analysis of an equivalent circuit for thecell injected with current and its neighboring cells in themiddle layer of the lobes showed that the resistances of theplasmalemma, tonoplast and junction between two cells were 11.1,4.7 and 0.56 M, respectively. Numerous plasmodesmata in thejunctional walls of the cells were found by electron microscopy.The low resistance of the junction between cells must be dueto the presence of the plasmodesmata which allows an electrotonictransmission of action potential from cell to cell. (Received January 23, 1982; Accepted April 13, 1982)     

Characterization of new strains of nonphotosynthetic mutants of Chlamydomonas reinhardtii II. Quinones and cytochromes b-559, b-563 and c-553 in twelve mutants having impaired Photosystem II function

Twelve new strains of nonphotosynthetic mutants of Chlamydomonasreinhardtii having impaired functioning of Photosystem II werestudied with respect to their quinone and chloroplastic cytochromecontents and to various photooxidation reactions of cytochromesb-559 and c-553. The quinones were analyzed by chromatography,cytochromes b-563 and c-553 were measured spectrophotometricallyafter solubilization by Triton X-100, and cytochrome b-559 wasstudied by means of low-temperature difference spectra. Noneof these mutants showed a great deficiency of plastoquinoneA, ubiquinone Q9, cytochrome b-563 or cytochrome c-553. Butall lacked an ascorbate-reducible pool of cytochrome b-559 photooxidizableat 77?K. In spite of this deficiency, five mutants (Fl 18, Fl29, Fl 47, Fl 50, Fl 59) showed an appreciable photooxidationof cytochrome b-559 in the presence of FCCP at room temperature.The other strains performed only weak cytochrome b-559 photooxidationin the presence of FCCP, DCMU and DBMIB or p-benzoquinone (Fl39, Fl 42, Fl 52, Fl 54, Fl 57, Fl 60); in the mutant Fl 33,no cytochrome photooxidation was observed. These results pointed out that the pool of ascorbate-reduciblecytochrome b-559 photooxidizable at 77?K is different from thepool photooxidizable in the presence of FCCP at room temperature. (Received February 8, 1979; )     

Pigment Composition of Chlorophyll-Protein Complexes Isolated from the Green Alga Bryopsis maxima

SDS-solubilized thylakoid membranes of Bryopsis maxima showeda similar pattern to those of higher plants in SDS-poIyacrylamidegel electrophoresis. Absorption spectra and pigment compositionof both CP1 and CPa bands were similar to those of higher plantsand other algae. Five bands containing chlorophyll (Chl) b weredivided into three categories; a group of major light-harvestingChl a/b-protein complexes (LHCP 1, LHCP 2 and LHCP 3), a minorLHCP (LHCP 3') and a photosystem I complex (CP1a). LHCP 1, thehigh molecular form, showed the lowest Chl a/b ratio among theLHCPs, and contained only xanthophylls as carotenoids. LHCP2, LHCP 3 and LHCP 3' bands contained xanthophylls and carotene.Carotenoid composition of LHCP 3' was different from that ofthe major LHCPs. CP1a band contained a considerable amount ofsiphonaxanthin and siphonein. (Received May 24, 1985; Accepted December 13, 1985)     

Changes in cytochrome contents and respiratory activity of Bacillus cereus Strain T during germination, vegetative growth and sporulation

Vegetative cells of Bacillus cereus Strain T contain cytochromeb-562, a minor b-type component, in addition to known components,cytochrome a+a₃, cytochrome b-557 and cytochrome c-551. Also,the spores contain low but definite amounts of cytochromes b-562and c-551, which were oxidized when the spores were shaken withair. Contents of cytochromes a, b and c per cell and per cellnitrogen, and the activity of glucose oxidation increased duringspore germination and elongation. During the stage precedingfirst cell division, cytochrome contents per cell increasedin parallel with the increase of cell nitrogen, while the activityof glucose oxidation decreased. During early exponential growth,the content of cytochrome b per cell nitrogen and respiratoryactivity with glucose again increased. When cells entered thesporulation stage, characterized by structural changes insidethe cells, the activity of glucose oxidation began to decrease,while that of acetate or succinate oxidation started to increase.During the sporulation process, the contents of the three cytochromecomponents continued to increase and reached the highest levelin cells containing completed spores, but the activity of respirationwith endogenous or added substrates was negligible in thesecells. (Received November 10, 1975; )     

The presence of chlorophyll b and the estimation of phaeopigments in marine phytoplankton

A reverse-phase h.p.l.c. technique was used to estimate theconcentration of chlorophyll b in phytoplankton cultures, fecalpellets of Calanus pacificus, and suspended paniculate matterfrom the Central North Pacific, Oregon coastal waters, and DabobBay (a temperate fjord in Puget Sound, WA, USA). The purposewas to assess the distribution of this pigment in the euphoticzone and its effect on the fluorometnc estimation of phaeopigments.Analyses of natural waters confirm high chlorophyll b concentrations(median mass ratio of b:a > 0.3) at the depth of the chlorophylla maximum in tropical waters while values for temperate planktonare relatively low (median mass ratio of chl b:a = 0.05) andpatchy. Zooplankton fecal pellets showed a significant enrichmentin chlorophyll b, suggesting grazing as a mechanism to explainhigh concentrations of this pigment at the bottom of the euphoticzone. It is estimated that the presence of chlorophyll b couldcause an average overestimation of phaeopigment concentrationby the fluorometnc technique of 38% between 0 and 200 m in theCentral North Pacific. This effect is more pronounced at thelayer of chlorophyll b maximum (120–140 m). ¹Present address: Marine Biology Research Division, A-002, ScrippsInstitution of Oceanography, La Jolla, CA 92093, USA     

Formation of Chlorophyll-Protein Complexes during Greening 1. Distribution of Newly Synthesized Chlorophyll among Apoproteins

The relationship between the accumulation of Chl and the apoproteinsof the light-harvesting Chl a/b-protein complex of PS II (LHCII)during the greening of cucumber cotyledons was studied. LHCIIapoproteins were not detected in etiolated cotyledons. Uponillumination, Chl a was formed as a result of photoconversionof protochlorophyllide (Pchlide) which had accumulated in thedark. During the lag period that preceded the accumulation ofChl, a small amount of LHCII apoproteins appeared. The amountof LHCII apoproteins increased with increases in levels of Chlb, though somewhat more rapidly during the first 10 h of greening.Treatment with benzyladenine (BA) or levulinic acid (LA) wasused to vary the supply of Chl a for apoproteins by promotingor inhibiting the synthesis of Chl a, respectively. LA decreasedbut BA increased the rate of accumulation of Chl b and LHCIIapoproteins. Only small amounts of Chl b and LHCII apoproteinswere formed under intermittent illumination. However, in thepresence of chloramphenicol (CAP), which inhibits the synthesisof plastome-coded proteins including apoproteins of the P700-Chla-protein complex (CP1) and a Chl a-protein complex of PS II(CPa), we observed the accumulation of Chl b and LHCII apoproteins,both of which are of nuclear origin. During incubation in thedark after intermittent exposure to light, CAP alone allowedneither destruction nor accumulation of Chl b and LHCII apoproteins,but it did enhance the effect of CaCl₂ in inducing both Chlb and these apoproteins. These results can be explained by assumingthat apoproteins of CP1 and CPa have a higher affinity for Chla than do LHCII apoproteins. When the availability of Chl ais limited, these apoproteins compete with one another for Chla, with the resultant preferential formation of CP1 and CPa.However, when the supply of Chl a becomes large enough for saturationof apoproteins of CP1 and CPa, some of the Chl a is incorporatedinto LHCII apoproteins either directly or after conversion toChl b. Thus, the formation of different Chl-protein complexes(CPs) is regulated by the relative rates of synthesis of Chla and apoproteins and by differential affinities of the apoproteinsfor Chl a. ⁴Present address: Kyowa Hakko Co., Ltd., 4041, Ami-machi, Inashiki,Ibaraki, 300-03 Japan (Received September 14, 1989; Accepted April 26, 1990)     

Isolation, Properties and a Possible Function of a Water-Soluble Chlorophyll a/b-Protein from Brussels Sprouts

A water-soluble Chl a/b-protein (CP673) was isolated and purifiedfrom Brussels sprouts (Brassica oleracea L. var. gemmifera DC).The protein had a molecular mass of 78 kDa and an isoelectricpoint of 4.7, consisted of three or four subunits of 22 kDaand was extremely heat-stable. Although CP673 contained aboutone Chl a per protein, the blue and red absorption bands ofChl a that consisted of three or four Chl a forms with differentabsorption maxima suggested that there are several differentmodes or sites of binding for Chl a. Chl a/b ratio of largerthan 10 also indicated that Chl b is present only in a smallfraction of CP673. The heterogeneity of CP673 in terms of compositionand binding of Chl suggests that Chl is not an intrinsic componentof the Chl-protein. Homology search showed that the N-terminalamino acid sequence of CP673 is highly homologous with thatof a 22 kDa protein that accumulates in water-stressed leavesof two Brassicaceae plants, rapeseed and radish, but not withthose of the light-harvesting Chl a/b-proteins of photosynthesis.A possible function of the water-soluble Chl-protein was discussed. (Received September 17, 1996; Accepted November 18, 1996)     

Properties of light-harvesting chlorophyll a/b-protein, and photosystem I chlorophyll a-protein, purified from digitonin extracts of spinach chloroplasts by isoelectrofocusing

A procedure for purifying both light-harvesting chlorophylla/b-protein and photosystem I chlorophyll -protein from digitoninextracts of spinach chloroplasts is described. This procedureuses isoelectrofocusing on Ampholine at the last step and permitsisolating all of the chlorophyll-proteins from the same extractin a better yield and a highly pure state. The purified light-harvesting chlorophyll a/b-protein whichhas an isoelectric point (pi) of 4.35 (?0.1) and a single polypeptideof 24 kilodaltons (kD), shows slightly higher chlorophyll a/Aratio of 1.35 than the values reported for the preparationsobtained by anionic detergents. This chlorophyll-protein exhibitsa markedly high and sharp fluorescence band at 681 nm at 77?Kwhich is not found on the chloroplast emission spectrum. Photosystem I chlorophyll a-protein focuses on Ampholine intotwo bands with pi values of 4.75 (?0.1) and 4.80 (?0.1). Thesetwo fractions show the same absorption spectra (maximum at 678nm at room temperature) and emission spectra (maximum at 734nm at 77?K) and have the same constituent polypeptides: onelarge band at 55–64 kD and six minor bands (21.5, 20,19, 18, 16 and 15 kD). The polypeptide composition and the P-700to chlorophyll a ratio (1 to ca. 80) of this preparation arevery similar to those of the photosystem I reaction center preparationobtained from Swiss chard chloroplasts by Bengis and Nelson(8). (Received October 31, 1978; )     

Chlorophyll a forms in mesophyll and bundle sheath chloroplasts of Zea mays leaves grown at low light intensity

Mesophyll and bundle sheath chloroplasts were prepared fromleaves of Zea mays grown at light intensities of 1.1 and 240µW/cm², respectively. The mesophyll chloroplasts thatdeveloped at the low intensity and bundle sheath chloroplatsthat developed at both low and high intensities showed higherratios of chlorophyll a/b and P700/chlorophylls compared withthe normal ratios found for the mesophyll chloroplasts thathad developed at the high intensity. Derivative absorption spectrophotometryat 77?K revealed that the low intensity mesophyll chloroplastscontained more of chlorophyll a forms with longer wavelengthred bands than high intensity mesophyll chloroplasts. More ofthe longer wavelength forms of chlorophyll a were also presentin the bundle sheath chloroplasts that had developed at lowand high intensities. All these four types of chloroplasts showedtwo peaks of fluorescence, one at 687 hra and the other at 733or 738 nm. In addition to these peaks, the high intensity mesophyllchloroplasts showed a shoulder at 697 nm, and the two typesof bundle sheath chloroplasts showed a shoulder at 680 nm. (Received June 17, 1974; )     

Growth of grana from "primary" thylakoids in Phaseolus vulgaris

The plastids of young dark-grown bean leaves, exposed to periodiclight are agranal, devoid of chlorophyll b and contain primarythylakoids and chlorophyll a. Transfer of these plants to continuousillumination results in synthesis of new chlorophyll a, chlorophyllb and grana. This study was done in order to study whether andhow the grana are formed from preexisting primary thylakoids.¹⁴C--aminolevulinic acid was used to label the chlorophyll aof the primary thylakoids, and its fate was studied after transferof the plants to continuous light. It was found that chlorophyll b and grana become ¹⁴C-labelled.The total radioactivity of chlorophyll b per bean increasedwith the parallel decrease of that of chlorophyll a. All subchloroplastfractions, obtained after digitonin disruption of chloroplasts,contained chlorophyll a of equal specific radioactivity. Thespecific radioactivity of chlorophyll b was lower than thatof chlorophyll a, and, in addition, it was lower in the granathan in the stroma lamellae fraction. The data suggest that chlorophyll b is formed from chlorophylla; the grana are formed by stacking of preexisting primary thylakoids;chlorophyll b is synthesized faster in the grana than the stromalamellae; the newly formed chlorophyll a molecules are distributedat random throughout the developing photosynthetic membraneand not on specific growing sites. (Received April 24, 1976; )     

Determination of chlorophylls and carotenoids of marine phytoplankton: separation of chlorophyll a from divinylchlorophyll a and zeaxanthin from lutein

A rapid reverse-phase HPLC method is presented for the identificationand quantification of most of the phytoplankton pigments. Thismethod yields the resolution of divinyl-chlorophyll a and chlorophylla, as well as the partial resolution of lutein and zeaxanthin,and of divinyl-chlorophyll b and chlorophyll b. In addition,chlorophylls c_1,2 and c₃ are well resolved. The analysis timefor one sample is 20 mm, which makes this method particularlysuited when large numbers of samples have to be processed.     

Temperature-dependent development in marine copepods: comments on choices of models

Guerrero et al (1994) promote Tauti's equation, rate = a exp(b temp), as simple to fit by log conversion (which may be formallyinappropriate), empirically adequate, and having ‘appropriatebiological characteristics’. No function is justifiablefrom reductionist theories, but Belehrádek's, rate =a (temp – T₀)^b, with b fixed for the taxon of interest,fits equally well, and singularly distinguishes differencesattributable to temperature adaptation (T₀, often misunderstoodas ‘biological zero’), and to size or other species-dependentproperties (a).     

Regulation of Electron Transport Composition in Cyanobacterial Photosynthetic System: Stoichiometry among Photosystem I and II Complexes and Their Light-Harvesting Antennae and Cytochrome b6/fComplex

This study was done to confirm our previous observation withthe pattern of changes in electron transport composition inducedby an imbalance of the electron transport state. Contents ofphotosystem (PS) I and II complexes and their antennae and Cytb₆/f complex were determined for systems of cyanobacterium SynechocystisPCC 6714 of different PS I/PS II ratios. The results indicatedthat (1) the observed changes in the PS I/PS II ratio are not-dueto regulation of the activities of the respective PS's but tochanges in their contents, (2) the molar ratio between PS IIand Cyt b₆/f complexes was fairly constant when marked changesoccurred in the PS I content, and (3) the PS II and Cyt b₆/fcontents per cell remained fairly constant while the PS I contentchanged markedly. These findings agree with our previous observationwith autotrophic cells of Anacystis nidulans Tx 20 and supportour argument that in cyanobacterial and red algal electron transportsystems, the content of the terminalcomponent(s), such as PSI complex, is regulated in order to maintain a balance betweenthe electron influx by PS II action to the system and the effluxby PS I action from it. (Received June 3, 1987; Accepted September 20, 1987)     

Purification and Characterization of Light-Harvesting Chlorophyll a/b-Protein Complexes of Photosystem II from the Green alga, Bryopsis maxima

Light-harvesting chlorophyll a/b-proteins of photosystem II(LHC II) were purified from thylakoid membranes of the greenalga, Bryopsis maxima. Extraction with digitonin did not solubilizechlorophylls (Chl) and carotenoids to any significant extent.Two forms of purified LHC II, P4 and P5, with respective apparentparticle sizes of 280 and 295 kDa, were obtained by sucrosedensity gradient centrifugation and column chromatography onDEAE-Toyopearl. P4 and P5 had similar spectral absorption at77 K with Chl a maxima at 674, 658 and 438 nm and Chl b maximaat 649 and 476 nm. Carotene was not present in P4 or P5. Fluorescenceexcitation spectra demonstrated that Chl b, siphonaxanthin andsiphonein can efficiently transfer absorbed light energy toChl a. P4 and P5 each contained two apoproteins of 28 and 32kDa, with similar but not identical amino acid compositions.P5 contained 6 molecules of Chl a, 8 of Chl b and 5 of xanthophyll(three molecules of siphonaxanthin and one each of siphoneinand neoxanthin) per polypeptide. (Received September 11, 1989; Accepted December 11, 1989)     

Effects of 5-Aminolevulinic Acid on the Accumulation of Chlorophyll b and Apoproteins of the Light-Harvesting Chlorophyll a/b-Protein Complex of Photosystem II

The effects were examined of 5-aminolevulinic acid (ALA) onthe accumulation of Chl and apoproteins of light-harvestingChl a/b-protein complex of photosystem II (LHCII) in cucumbercotyledons under intermittent light. A supply of ALA preferentiallyincreased the accumulation of Chl a during intermittent illumination.However, when cotyledons were pretreated with a brief exposureto light or benzyladenine (BA), the stimulatory effect of ALAon the increase in the level of Chl b was greater than thatin the level of Chl a, resulting in decreased ratios of Chla/b. Time-course experiments with preilluminated cotyledonsrevealed that LHCII apoproteins accumulated rapidly within thefirst 30 min of intermittent illumination with a decline duringsubsequent incubation in darkness. A supply of ALA did not affectthe accumulation of LHCII apoproteins during the intermittentlight period, but it efficiently inhibited the decline in theirlevels during the subsequent darkness. After exposure to a singlepulse of light of BA-treated cotyledons, the prompt increasein levels of LHCII apoproteins was not accompanied by the formationof Ch b, which began to accumulate later. The pattern of changesin levels of LHCII apoproteins was quite similar to that inlevels of Chl a. These results suggest that LHCII apoproteinsare first stabilized by binding with Chl a and that an increasedsupply of Chl a and the accumulation of LHCII apoproteins areprerequisites for the formation of Chl b. ¹Present address: Department of Chemistry, Faculty of Scienceand Technology, Meijo University, Aichi, 468 Japan.