Purification and properties of the alkaline phosphatase ofSerratia marcescens

1. A hypothesis based on the Hill-Bendall-model of photosynthetic electron transport is proposed to explain positive and negative photo-phobotaxis inPhormidium uncinatum. In the non-cyclic electron chain a pool is located into which photosystem II (e. g. by absorption by C-phycoerythrin, 561 nm) feeds electrons while photosystem I (e.g. 723 nm) drains electrons out of it.
2. Interruption of the electron flow into the pool causes a sudden decrease of the pool size and thus a positive phobic response. This happens e.g. when an organism leaves a trap which is illuminated by a wavelength absorbed by photosystem II pigments (e. g. 561 nm).
3. A negative reaction takes place when electrons are suddenly drained out of the pool; again the pool size decreases. This is the case when an organism enters a light trap illuminated by photosystem I light (723 nm).
4. The net flow of electrons into or out of the pool—and thus the reaction sense—can be manipulated by the relative excitation of the two photosystems or by blocking the electron influx by DCMU.

     

Effect of external factors on phototaxis of Chlamydomonas reinhardtii

1. A fully automated phototaxis monitoring device is described for measuring photo-topatactic responses of flagellated organisms.
2. Photokinesis can be demonstrated in Chlamydomonas cells only after a dark period of about 72 hrs.
3. Pre-darkening of a few hours duration raises the phototactic disposition, whereas pre-illumination has no significant effect.
4. Circadian rhythms can be initiated by only one period of darkness or lower light intensity, whereas a period of higher intensity does not induce rhythms. The period length of the circadian rhythms is about 24 hrs.

     

Experimental problems using electronic particle counters

The following scheme lists the problems which have been and will continue to be encountered in grazing experiments on natural particle assemblages. In some cases there are solutions, but many of the problems listed below remain intractable. To solve them will require innovative approaches, most probably combining the use of particle counters with other techniques to approach the problem to be solved from several angles at once. Variance between samples due to

- statistical causes (especially caused by large particles) and

- differences between experimental bottles and between experimentals and controls at start of experiment. Shifts in particle distribution during experiment due to

- increase in small particles

- growth of bacteria and other organisms due to excretion of grazers and

- breakage during handling by grazers into fragments. Processes, independent of grazing, leading to changes in particle size distribution due to

- primary production

- grazing by additional components and

- bacterial growth on detrital particles and the formation of detrital flocs.

     

Extracellular and intracellular determination of light-induced potential changes during photophobic reactions in blue-green algae

Light-induced potential changes have been measured in the filamentous blue-green alga Phormidium uncinatum both intracellularly and between the two ends of a trichome. There is evidence that these potential changes are correlated with photophobic reactions in this organism.

1. The potential changes follow the light-dark regime with a lag phase of about 10 s. The photophobic reaction time has been found to be about the same length of time.
2. The action spectra of both externally and internally measured light-induced potential changes correspond with the photophobic action spectrum, indicating the participation of the main photosynthetic pigments of Phormidium, chlorophyll a and phycobilins.

A hypothesis is being discussed according to which sensory transduction between photoreceptor and motor apparatus of the cell is mediated by light-induced electrical potential changes.     

Triplet-minus-singlet absorbance difference spectra of reaction centers of Rhodopseudomonas sphaeroides R-26 in the temperature range 24–290 K measured by Magneto-Optical Difference Spectroscopy (MODS)

The recently developed technique of Magneto-Optical Difference Spectroscopy (MODS) [10] has been applied to reaction centers (RC) of the photosynthetic bacterium Rhodopseudomonas sphaeroides R-26. Absorbance changes induced by a magnetic field are measured as a function of wavelength yielding the triplet-minus-singlet (T-S) absorbance difference spectrum. (T-S) spectra thus obtained have been measured from 24–290 K. Going from low to high temperature the (T-S) spectra show the following features:

1. A rapid decrease of positive absorption bands at 809 and 819 nm.
2. A slow appearance of a band shift at 798 nm.
3. A shift of the peak wavelength of the Q_y absorbance band of the primary donor P-860 from 992 to 861 nm, and of its Q_x band from 603 to 600 nm.

The spectra at 24, 66, 116, and 290 K have been analyzed by Gaussian deconvolution. The 800 nm region of the spectrum at 24 K can be decomposed in a combination of two band shifts and an appearing band. The temperature dependence of the spectra in this region is well explained by spectral broadening of the two shifting bands combined with a decrease in intensity of the appearing band when the temperature increases. The two shifting bands in the 800 nm region are identified as the two bands at 803 and 813 nm which together make up the 800 nm band in the absorption spectrum and are assigned to the two accessory RC bacteriochlorophylls (BChls). The band shift of the 813 nm pigment is appreciably larger than that of the 803 nm pigment. The appearing band at 808 nm is attributed to monomeric absorption of ³P-860, the triplet state being localized on one BChl. We find no evidence for admixture of a charge transfer (CT) state of ³P-860 with one of the accessory BChls at higher temperature.     

Rhythmicity of chromophore turnover of visual pigment in the Antarctic amphipod Orchomene plebs (Crustacea; Amphipoda)

1. Relative retinal amounts in the compound eye of the Antarctic amphipod Orchomene plebs were assessed during conditions of continuous summer daylight every 3 h over a period of 48 h. The habitat of the experimental animal is the bottom of the Ross Sea (78°S; 166°E) down to depths of at least 400 m; water temperature is a constant — 1.8° C. A periodicity of 12 h was detected with relative amounts of 11-cis retinal exhibiting peaks at midday and at midnight and troughs at 7.00 h and 19.00 h.
2. The result that 90% of retinoid were insoluble in n-hexane suggests that at least 90% of the measured retinoid were attached to membrane-bound proteins such as opsin.
3. Selective light adaptation showed that the visual pigments were thermostable and photoregenerable. The main absorbance peak of rhodopsin, compared with metarhodopsin, seems to be in the longer wavelengths.

     

Morphological and physiological properties of pheromone-triggered flipflopping descending interneurons of the male silkworm moth,Bombyx mori

1. The morphology of descending interneurons (DNs) which have arborizations in the lateral accessory lobe (LAL) of the protocerebrum, the higher order olfactory center, and have an axon in the ventral nerve cord (VNC), were characterized in the male silkworm moth, Bombyx mori.
2. Two clusters (group I, group II) of DNs which have arborizations mainly in the LALs were morphologically characterized. The axons of these DNs are restricted to the dorsal part of the each connective (Figs. 1–5).
3. Pheromonal responses of the group I and group II DNs were characterized. Flipflopping activity patterns, which have two distinct firing frequencies (high and low) in response to sequential pheromonal stimulation, were usually recorded (Figs.6–10).
4. Two types of flipflopping activity patterns were classified into those that had an antiphasic relationship (called the ‘FF’ type) between the left and right connectives and those with a synchronized relationship (‘ff’ type) (Figs. 8–12). We propose that some group II DNs show ‘FF’ flipflopping activity patterns (Fig. 10).
5. A state transition was usually elicited by less than 10 ng bombykol, the principal pheromone component. Extra impulses were elicited during constant light stimulation (Fig. 9).
6. Our results suggest that the LAL olfactory pathways might be important for producing flipflopping activity patterns (Fig. 11).

     

Photo-activation of respiration in the presence of CO in sperm of several marine invertebrates

• 1.1. The light irradiation at examined wavelengths between 360 and 630 nm enhanced the respiratory rate in the presence of CO in sperm, as well as eggs, of the sea urchins, Anthocidaris crassispina and Hemicentrotus pulcherrimus, the starfish, Asterina pectinifera, and the echiuroid, Urechis unicinctus.
• 2.2. The maximum peaks of stimulating effects of light irradiation on CO-insensitive respiration were found at wavelengths of 430, 530 and 570 nm in sperm of these species.
• 3.3. The respiration in the presence of CO was insensitive to light irradiation in sperm and eggs of the oyster, Crassostrea gigas and the tunicate, Ciona intestinaris.

     

Seasonal variations in biochemical constituents of the clam,Paphia laterisulca

1. The total protein, fat and glycogen contents were estimated from the edible clam, P. laterisulca. Seasonal variations in these constituents along with the water content were studied.
2. The gonad index in P. laterisulca was found to increase during the ripe condition and in winter (December–January) and decrease on spawning.
3. A relatively high water content was obtained during monsoon (June to September). This might be due to the loss of salts and gain of water in low salinities.
4. Protein content varied with the reproductive cycle of the clam. The level reached its peak in the mature stage and declined on spawning. Immature clams showed less protein content than gravid ones.
5. Lipid content started to increase as the gametogenesis commenced, reached its peak in fully mature condition (August) and sharply declined due to the shedding of gametes during spawning.
6. Glycogen content was high during the period of active gameto-genesis (May–June). A sharp decrease took place when the clams were fully ripe (July). The glycogen might have been utilized in the formation of active ripe gametes.
7. After starvation for twelve days, total protein and fat contents remained constant, while glycogen content decreased by 66.82%. The water content increased by 4.67%.
8. Seasonal variation in the organic constituents are discussed in relation to the reproductive cycle of the clam.

     

Comparative kinetic light-scattering and -absorption photometry

A recent development of kinetic light-scattering and -absorption photometry is described. Essential points are:

1. In the scattering experiment, amplitude resolution of 2 · 10^?5 (single flash) by application of a differential detector, stability of the same magnitude due to optical compensation, high intensity at the detector due to special optics for the scattered light and semiconductor sources.
2. In the absorption measurement, elimination of scattering contributions by the dual wavelength-method and by high aperture optics.
3. Simultaneous measurement of absorption and scattering. The application of the method is described in using signals from isolated bovine rod outer segments. A reliable procedure is described by the use of which the originally measured light-scattering effects can be split up into single signals.

The method allows comparative kinetic analysis of absorption and scattering signals. The possible causal connections between pigment and membrane structure processes can be selected.     

A study on the rate of water transport of the clam katelysia opima in relation to environmental conditions

1. The neutral red technique was employed to study the rate of filtration in Katelysia opima.
2. The weight specific water filtration was found to be greater for younger clams compared to the older ones.
3. The rate of water filtration increased with decreasing salinity.
4. Water filtration was found to increase as temperature increased, reaching a maximum at 35°C. but then sharply decreasing at 39°C.
5. Light had no significant effect on the rate of filtration.
6. Suspended matter was found to affect the rate of water filtration.
7. The rate of filtration was low at high pH and high in low pH.
8. The rate of water filtration was found to be faster during high tide than during low tide.
9. The presence of the parasitic crab, Pennotheris sp., in the mantle cavity of clams had a marked effect on the particle filtration.
10. Accidental cut of the siphon tips had no effect on the rate of filtration.

     

Effects of energy deprivation on the fly pupil mechanism: evidence for a rigor state

The energy dependence of the pupil pigment-migrations in the fly Musca domestica was studied in live animals, using optical techniques and nitrogen-gas induced anoxia. The results obtained can be summarized in 3 points:

1. Energy deficiency can make the pupil mechanism stop in any state, extreme or intermediate.
2. Anoxia induced during intermittent stimulation makes the pupil stop in the closed state (aggregated pigment granules).
3. During long-term anoxia the pupil very slowly opens (dispersal of pigment granules), irrespective of ambient intensity.

The slow anoxic opening (point 3) is more than 1000 times slower than that predicted for free diffusion of pigment granules in water. Assuming realistic values of cytoplasm viscosity, this implies that anoxia causes the pigment granules to attach to rigid structures in the cells, in analogy with the rigor state in anoxic muscles. The rigor phenomenon in the pupil mechanism prevents experimental discrimination between active and passive processes of pigment migration. Normal pupil opening has a time course which agrees reasonably with a passive diffusion process, but it is argued that an active transportation of granules away from the rhabdom is more likely in the dark adapted eye.     

The use of a continuous assay system to show the stimulation of phosphoenolpyruvate production in intact mitochondria by valinomycin and by Mn2+

1. A method for the direct recording of the PEP efflux from isolated mitochondria is described.
2. This method has been used to show the stimulation of PEP efflux by externally added Mn⁺⁺ ions.
3. Valinomycin, uncoupler and oleate were also shown to stimulate PEP efflux.
4. Valinomycin caused an increase in the internal concentration of both PEP and citrate.
5. The results indicate that the major pathway of PEP synthesis in isolated mitochondria is via PEP carboxykinase and the results do not call for an unknown pathway of metabolism.
6. Two interactions between PEP and citrate are described; competition for the mitochondrial interior and the stimulation of PEP production by citrate.

     

The use of toxicokinetics for the safety assessment of drugs acting in the brain

Pharmacological and toxicological studies undertaken on drugs that affect the brain are frequently performed in disparate species under various experimental conditions, at doses often greatly in excess of those expected to be administered to humans, and the findings are extrapolated implicitly or explicitly with scant regard to differences in the biodisposition of the drugs. Such considerations are necessary since:

1. Species;
2. Strain;
3. Gender;
4. Route;
5. Dose;
6. Frequency and time of administration;
7. Temperature;
8. Coadministration of drugs; and
9. Surgical manipulation

are but some of the factors that have been shown to influence the kinetics and metabolism of drugs. This article, using MDMA and other phenylethylamines as examples, provides evidence for the need to measure the exoosure of the drugs and their active metabolites in blood and brain (toxicokinetics) in order that conclusions based only on dynamic, biochemical, or histological evidence are more pertinent. Further, the combined use of toxicokinetic-dynamic modeling can lead to a better appreciation of the mechanisms involved and a more useful approach to the calculation of safety margins.     

Improvements of the membrane filter method for DNA:rRNA hybridization

We describe and recommend the following improvements of DNA:rRNA membrane filter hybridization methods. One of our aims was to avoid DNA release from filter discs during hybridization.

1. Our hybridization conditions are 2 SSC in aq. dest., with 20% formamide, 50 C, overnight for 16 hr.
2. Duplexing is over in 8–10 hr.
3. Formamide has to be very pure (O.D.≤0.2/cm light path at 270 nm).
4. RNAase treatment: 250 μg/5 ml 2 SSC/filter at 37 C for 1 hr.
5. Our conditions for stepwise thermal denaturation are: 5°C steps from 50C to 90C in 1.5 SSC in 20% formamide.
6. Single-stranded DNA, fixed on membrane filters, and stored in vacuo at 4C, can be used reliably for hybridization for up to 20 months.
7. Concentrated DNA in 0.1 SSC, quick-frozen at ?50 C and stored at ?90 C for up to 2 years can be used for hybridization without much change.
8. A CsCl gradient purification step yields much purer DNA, but increases the release of DNA from filters by about 20%. Filters with 20% more DNA is a compensation.
9. rRNA can be stored for 20 months in SSC or 2 SSC at ?12C without changing the hybridization results.

     

Luminescence and thermoconductive properties of the oriental hornet (Vespa orientalis) cocoon: Evidence for photoinduced electron-transfer system

• 1.1. The luminescence properties of the cocoon spun by the larvae of the Oriental hornet was studied.
• 2.2. When light was shined upon the cocoon caps with excitation wavelengths of 240, 290 or 312 nm, a luminescence occurs where maximal intensity is at a wavelength of about 360 nm.
• 3.3. The luminescence is dependent on (a) the excitation wavelength, (b) the side of the cocoon irradiated and (c) on the age of pupating larva.
• 4.4. The cocoon cap displays electric conductance properties, in that the resistance to current is temperature dependent. Evidence is provided for a photoinduced electron-transfer system.

     

Growth requirements of blue-green algae under blue light conditions

1. Growth requirements of blue-green algae containing only the c-phycocyanin + chlorophyll a pigment system have been studied under blue light (380–540 nm) which approximates light conditions existing in subsurface waters in nature.
2. While a few species were capable of very slow photosynthetic growth on minimal medium with NO₃ ^- as nitrogen source, most species were dependent on organic compounds for comparable growth under this condition. Some organisms did quite well with only Casamino Acids as a supplement, others did well with only glucose. One species, Agmenellum quadruplicatum strain PR-6, grew only when glucose and Casamino Acids were supplied simultaneously.
3. Inhibitory effects of blue light on CO₂ fixation and nitrogen metabolism are noted as possible explanations of these responses.

     

Seasonal variation in the biochemical composition of Mytilus viridis at Ratnagiri on the West Coast of India

1. The seasonal variation in the water, protein, fat and glycogen contents of the mussel, Mytilus viridis has been studied for the year March, 1974 to March, 1975.
2. The water level increased during the monsoon season and decreased in summer.
3. The level of protein, fat and glycogen showed correlation with the reproductive cycle of the mussel.
4. The protein level was high when the mussels were mature and dropped during the breeding period.
5. During sex change from male to female in May the protein level remained high whereas during sex change from female to male in October and November it was low.
6. The fat level was high in mature mussels and declined on spawning.
7. The glycogen level was at its peak in immature mussels and low in mature.

     

Time course of the Houseflies' landing response

The landing response of stationary flying houseflies Musca domestica has been recorded on video tape. The leg movements were quantitatively evaluated. It could be demonstrated that:

1. only the first two pairs of legs are involved in the reaction (Fig. 1). Prothoracic tarsi are lifted beyond the head, mesothoracic tarsi are lowered and moved sidewards (Fig. 2a and b).
2. the movement of the tarsal tips is mainly due to an opening of one single joint per leg, i.e. the femurtibia joint of the prothoracic leg (Fig. 2c), and the coxa-femur joint of the mesothoracic leg.
3. the landing reaction is a fixed action pattern which does not seem to require further sensory input once it is released (Fig. 4d).
4. the landing responses to a light-off stimulus and to expanding patterns with different angular velocities are indistinguishable (compare Fig. 3a-c with Fig. 2a-c). The only parameter that is obviously dependent on the stimulus conditions, is the latency of the reaction (Fig. 4a-c).

     

Effects of scaphognathite nerve stimulation on the acutely deafferented crab ventilatory central pattern generator

1. Sensory axons from crab (Carcinus maenas) scaphognathites enter the thoracic ganglion primarily via the LNb branch of the levator nerve. The LNa branch of the levator nerve and the depressor nerve each contain relatively few sensory axons.
2. Acutely deafferented ventilatory central pattern generators show a free running burst rate which is lower than that observed in intact crabs. Electrical stimulation of the levator nerve, or of its LNb branch, increases the burst rate in a frequency dependent manner. Stimulation at high enough intensity to recruit afferents will restart a paused motor rhythm. Stimulation of the levator nerve with short pulse trains phase resets and can entrain the rhythm.
3. In addition to increasing the burst rate, LNb stimulation also causes a progressive elimination of motor neurons from the bursts as the stimulating frequency increases, probably due to depolarization of the 3 oval organ ‘giant’ afferent axons in this branch. Intracellular depolarization of single oval organ afferents will also inhibit some motor neurons as well as slow or stop the rhythm.
4. Continuous stimulation of the depressor nerve does not affect the ganglionic burst rate and this nerve contains only a few small diameter afferent axons; however, brief trains of stimuli can reset the rhythm in a phase-dependent manner.