ELECTRICAL VARIATIONS DUE TO MECHANICAL TRANSMISSION OF STIMULI

Mechanical stimulation of Nitella often produces responses resembling propagated negative variations but traveling faster and going past a killed spot. They appear to result from a mechanical disturbance traveling along the cell and stimulating each spot it touches (i.e. the stimulus itself travels). They are called mechanical variations to distinguish them from propagated negative variations. A mechanical disturbance may cause an irreversible change (death wave), but in traveling along the cell it may lose intensity and then produce only a reversible response (mechanical variation) which may eventually change to a propagated negative variation. The all or none law does not apply to incomplete mechanical variations, for the response varies with the strength of the stimulus.     

Distant effects of toxic agents

Toxic solutions applied at one end of a Nitella cell 6 cm. long may produce little or no visible change in the structure of the protoplasm at the place of application but if the opposite end is covered with water its protoplasm soon disintegrates. If the middle of the cell is covered with mineral oil this region remains normal in appearance for half an hour or more. The result is due to the movement of substances in the cell. The loss of substances at the end where the toxic agent is applied results in loss at the opposite end if it is covered with water since water enters and travels along inside the cell carrying substances with it. This causes injury at the spot where the water enters. The conception developed here differs fundamentally from the usual view that the effects of injury spread gradually from the region where the toxic agent is applied to the immediately adjoining regions and thence to more remote places. The change produced by loss of substances produces an interesting pattern which deserves study.     

THE DEATH WAVE IN NITELLA : I. APPLICATIONS OF LIKE SOLUTIONS.

Experiments on cutting confirm the prediction that the current of injury will be positive when the cell is in contact with concentrated solutions and negative with dilute solutions. They support the idea that the protoplasm is made up of layers differing considerably in their properties, each having a death curve of simple and regular form, the more rapid alteration of the outer layer making the protoplasm more positive and the more rapid alteration of the inner making it more negative. From the point where the cell is cut a wave of some sort, which we may for convenience call a death wave, passes along the cell, setting up at each point it touches a death process which has the greater speed and intensity the nearer it is to the cut.     

A cell surface differentiation antigen of Drosophila

A monoclonal antibody, generated by immunization with gastrula stage Drosophila melanogaster embryonic cells, recognizes a cell surface antigen which shows tissue and stage specificity. The antigen appears for the first time during cellularization of the blastoderm embryo and is present on all cells until around 12 hr of development. It becomes progressively restricted to specific tissues during the second half of embryogenesis. By the time of hatching, only the nervous system, germ cells, and imaginal cells are positive. During metamorphosis differentiating imaginal tissues become negative so that in the adult only the nervous system and undifferentiated germ cells are positive, with gonadal sheaths showing some staining. A third wave of antigen loss occurs during gametogenesis, resulting in negative staining on the mature sperm and oocyte. All positive tissues appear to contain the same 63-kDa cell surface antigen. The antigen behaves as a general differentiation marker lost by tissues as they approach their terminal differentiated state. The nervous system and possibly gonadal sheaths may be exceptions to this general behavior.     

The mouse spot test. Evaluation of its performance in identifying chemical mutagens and carcinogens

The published results on 60 chemicals and X-rays investigated in the mouse spot test were compared with data on the same chemicals tested in the bacterial mutation assay (Ames test) and lifetime rodent bioassays. The performance of the spot test as an in vivo complementary assay to the in vitro bacterial mutagenesis test reveals that of 60 agents, 38 were positive in both systems, 6 were positive only in the spot test, 10 were positive only in the bacterial test and 6 were negative in both assays. The spot test was also considered as a predictor of carcinogenesis; 45 chemicals were carcinogenic of which 35 were detected as positive by the spot test and 3 out of 6 non-carcinogens were correctly identified as negative. If the results are regarded in sequence, i.e. that a positive result in a bacterial mutagenicity test reveals potential that may or may not be realized in vivo, then 48 chemicals were mutagenic in the bacterial mutation assay of which 38 were active in the spot test and 31 were confirmed as carcinogens in bioassays. 12 chemicals were non-mutagenic to bacteria of which 6 gave positive responses in the spot test and 5 were confirmed as carcinogens. These results provide strong evidence that the mouse coat spot test is an effective complementary test to the bacterial mutagenesis assay for the detection of genotoxic chemicals and as a confirmatory test for the identification of carcinogens. The main deficiency at present is the paucity of data from the testing of non-carcinogens. With further development and improvement of the test it is probable that the predictive performance of the assay in identifying carcinogens should improve, since many of the false negative responses may be due to inadequate testing.     

Photoelectrolysis using chlorophyll electrodes

By the conversion of sunlight to chemical energy, photoelectrolysis was carried out using two different chlorophyll-redox compound lining electrodes. These electrodes were prepared by covering platinum plates with chlorophyll and naphthoquinone or anthrahydroquinone and a conducting adhesive. These electrodes exhibit a photoexciting property. The potential was found to shift to a less noble state when the system of the chlorophyll-naphthoquinone electrode was inserted into NAD solution with illumination. On the other hand, the photoexcitation of the system of the chlorophyll-anthrahydroquinone electrode inserted into ferrocyanide solution made the potential more noble. (If the potential in the dark is in the positive region of the scheme

and the potential moves in the positive direction when the light is turned on, it can be said to be more positive or more noble. But if the potential moves in the negative direction in the light, but remains in the positive region, it can be said to become less noble, but it is not suitable to say that it becomes more negative.) To make the potential difference between two electrodes as big as possible, various factors such as intensity of illumination, molar ratio of chlorophyll to naphthoquinone or anthrahydroquinone and concentration of redox compound in electrolyte were examined.A cell was set up by combining the system of the chlorophyll-naphthoquinone electrode in NAD solution with that of the chlorophyll-anthrahydroquinone electrode in ferrocyanide solution, and photoelectrolysis was carried out by closing the external circuit with illumination. The photovoltage between the two electrodes was 0.25 V and the current density was 8 μA/cm². It was found that NAD was reduced to form NADH at the chlorophyll-naphthoquinone electrode and ferrocyanide was oxidized to form ferricyanide at the chlorophyll-anthrahydroquinone electrode.     

Conductance Change Associated with Receptor Potentials of Gustatory Cells in Rat

The electrical properties of gustatory cells and cells which do not respond to chemical stimuli in the taste bud of fungiform papillae in rats were studied by means of intracellular microelectrodes. Neither of these cell types showed spike electrogenesis. Gustatory cells showed a depolarization, the receptor potential, associated with an increase in the membrane conductance in response to NaCl, sucrose, and HCl, whereas quinine produced a decrease in the conductance together with an increase in the receptor potential magnitude. The reversal point of the receptor potential in response to NaCl or KCl was close to zero membrane potential, but in the case of quinine it was at a more negative potential level than the resting potential. From these results two receptive processes are postulated in the gustatory cell membrane. When the gustatory cells were stimulated for a long duration by concentrated NaCl or sucrose, receptor potentials showed adaptation with decrease in magnitude, but adaptation of the responses to HCl and quinine were hardly detected. Adaptation of the receptor potential was not correlated with conductance change.     

MEK drives cyclin D1 hyperelevation during geroconversion

When the cell cycle becomes arrested, MTOR (mechanistic Target of Rapamycin) converts reversible arrest into senescence (geroconversion). Hyperexpression of cyclin D1 is a universal marker of senescence along with hypertrophy, beta-Gal staining and loss of replicative/regenerative potential (RP), namely, the ability to restart proliferation when the cell cycle is released. Inhibition of MTOR decelerates geroconversion, although only partially decreases cyclin D1. Here we show that in p21- and p16-induced senescence, inhibitors of mitogen-activated/extracellular signal-regulated kinase (MEK) (U0126, PD184352 and siRNA) completely prevented cyclin D1 accumulation, making it undetectable. We also used MEL10 cells in which MEK inhibitors do not inhibit MTOR. In such cells, U0126 by itself induced senescence that was remarkably cyclin D1 negative. In contrast, inhibition of cyclin-dependent kinase (CDK) 4/6 by PD0332991 caused cyclin D1-positive senescence in MEL10 cells. Both types of senescence were suppressed by rapamycin, converting it into reversible arrest. We confirmed that the inhibitor of CDK4/6 caused cyclin D1 positive senescence in normal RPE cells, whereas U0126 prevented cyclin D1 expression. Elimination of cyclin D1 by siRNA did not prevent other markers of senescence that are consistent with the lack of its effect on MTOR. Our data confirmed that a mere inhibition of the cell cycle was sufficient to cause senescence, providing MTOR was active, and inhibition of MEK partially inhibited MTOR in a cell-type-dependent manner. Second, hallmarks of senescence may be dissociated, and hyperelevated cyclin D1, a marker of hyperactivation of senescent cells, did not necessarily determine other markers of senescence. Third, inhibition of MEK was sufficient to eliminate cyclin D1, regardless of MTOR.     

How is extinction risk related to population-size variability over time? A family of models for species with repeated extinction and immigration

It is well known that for an isolated population, the probability of extinction is positively related to population size variation: more variation is associated with more extinction. What, then, is the relation of extinction to population size variation for a population embedded in a metapopulation and subjected to repeated extinction and recolonization? In this case, the extinction risk can be measured by the extinction rate, the frequency at which local extinction occurs. Using several population dynamics models with immigration, we find, in general, a negative correlation between extinction and variation. More precisely, with increasing length of the time series, an initially negative regression coefficient first becomes more negative, then becomes less negative, and eventually attains positive values before decreasing again to 0. This pattern holds under substantial variation in values of parameters representing species and environmental properties. It is also rather robust to census interval length and the fraction of missed individuals but fails to hold for high thresholds (population size values below which extinction is deemed to occur) when quasi extinction rather than true extinction is represented. The few departures from the initial negative correlation correspond to populations at risk: low growth rate or frequent catastrophes.     

Electrochemical titrations of a ferredoxin-ferredoxin:NADP+ oxidoreductase complex

Potentiometric titrations employing an electrochemical thin-layer cell indicate that complex formation between ferredoxin and ferredoxin:NADP⁺ oxidoreductase alters the midpoint oxidation-reduction potentials of both proteins. The midpoint potential of ferredoxin in the complex becomes 22 ± 6 mV more negative compared to ferredoxin alone while the midpoint potential of ferredoxin:NADP⁺ oxidoreductase becomes 23 ± 4 mV more positive on complex formation.     

Electrical Characteristics of Insect Mechanoreceptors

External direct coupled recordings from the neurons of the mechanosensory hairs of insects show nerve impulses and graded slow potentials in response to deformation of the hair. These slow potentials or receptor potentials are negative going, vary directly with the magnitude of the stimulus, and show no overshoot when returning to baseline. The impulses have an initial positive phase which varies in size directly with the amplitude of the receptor potential. The receptor potential is related to the generator potential for the impulse in that it must attain some critical level before impulses are produced, and the frequency of impulses varies directly with amplitude of the receptor potential. The receptor potential does not return to the baseline after each impulse. In some receptors static deformation of the hair will maintain the receptor potential. It appears likely that both the receptor potential and the variation in size of the impulses are caused by a change in conductance of the cell membrane at the receptor site, and that the receptor potential originates at a site which is not invaded by the propagated impulses.     

Antigen as a positive and negative regulator of proliferation in cytotoxic lymphocytes. A model for the differential regulation of proliferation and lytic activity

The goal of these experiments has been to capitalize on the functionally distinct responses of heteroclitic CTL to cross-reactive Ag to explore the role of Ag in regulating CTL proliferation and lytic function. The experiments here have examined one of these clones in detail but several such clones appear similar. The principal findings were: 1) proliferation and lysis may be optimally stimulated by quantitatively different interactions, 2) Ag can have negative as well as positive effects on the proliferative potential of the responding cell, and 3) both positive and negative effects on lysis can be mimicked by agents which are similar to the second messengers produced by stimulating the TCR.     

The adsorption of DNA in the mercury electrolyte interface. 3. Reaction of DNA in the mercury-electrolyte interface

The adsorption of deoxyribonucleic acid in the mercury-electrolyte interface was investigated. The effect of this adsorption on the differential capacity of the electrical double layer at the interface between a stationary mercury drop electrode (HMDE) and a buffered aqueous sodium chloride solution was measured. The dependences of this differential capacity on potential, time, and pH was studied in the presence of native and also of denatured DNA. These results were compared with the adsorption of model compounds and with the general theory of the adsorption of polymers. The structure of the adsorbed DNA molecules corresponds to an alternating arrangement of two-dimensional, totally adsorbed sequences and three-dimensional loops extending into the solution. The adsorbed sequences and loops consist of several segments with a specific free-energy change of adsorption. Essentially this energy determines the distribution of the segments between adsorbed sequences and loops. The absolute value of this energy change per segment is fairly large in the case of negatively charged poly-electrolyte DNA at the weakly positively charged interface near the electrocapillary maximum (ECM). The fraction of totally adsorbed segments is relatively large in this potential region. The more negative the potential the lower is the absolute value of free energy change of adsorption per segment. Under the conditions unfavorable for the adsorption, only a few segments can be adsorbed. Most of the segments of the adsorbed DNA molecules extend into the solution and therefore fairly high interface concentrations can be reached. Thus, the arrangement of DNA molecules in the electrode surface is changed when the potential is altered from values near the ECM to more negative ones. This change should produce the wave on the differential capacity curves at a little more negative potential than that of ECM. At a more negative potential, intermolecular interactions between the loops extending into the solution may occur. The adsorption tendency of the resulting associates is higher than that of the isolated molecules. Therefore the isolated molecules desorb at sufficient negatively charged interface producing a round wave while the associates stay adsorbed. At this potential it is impossible for native DNA to generate associates because they are formed from the isolated molecules. This explains the hysteresis loop of the curves of differential capacity vs. potential by using the HMDE. The desorption of the associates is indicated by a sharp wave at much more negative potential. For denatured DNA the associates arise from the very few isolated adsorbed molecules at this potential; therefore, no hysteresis loop occurs. The association constant of denatured DNA must be much higher than that of the native DNA. The reasons for this are discussed.     

Visual evoked potentials produced by monocular flash stimuli in the cerebral cortex of the rabbit. I. Typography

The visually evoked potentials in the hemisphere contralateral to the stimulated eye in rabbit, can be described topographically as follows. While a positive wave (P1) begins forming in the anterior zones and in the V I binocular zone, the N0 wave, at times very large, is produced in a more occipital zone, which corresponds to the visual streak. Immediately afterwards, the positivity, P1, practically invades the whole of the hemisphere. After this, the N1 wave which is produced in the most posterior parts of the V I, begins forming. The whole phenomenon comes to an end when the P2 wave is generated in the most occipital zones.     

Effect of Long Chain Fatty Acids on Bacterial Respiration and Amino Acid Uptake

S ummary . Long chain fatty acids stimulated oxygen uptake by Gram positive bacteria at bactericidal and protoplast lytic concentrations and produced inhibition at higher levels. The order of activity between individual acids and effects of reversal agents on respiratory activity corresponded to those which produced bactericidal activity. Protoplasts were more susceptible to inhibition than whole cells. Gram negative bacteria were inhibited to a limited extent at high fatty acid concentrations, but spheroplasts were highly sensitive. Fatty acids inhibited amino acid uptake both aerobically and anaerobically at sub-bactericidal levels. The effects were reversed by metal cations, and reflected the activity of dinitrophenol and sodium azide. The susceptibility of organisms to inhibition was of the same order as the sensitivity to other antibacterial effects. The probable mode of action of the fatty acids is discussed in terms of the interference with energy metabolism within the bacterial cell.     

Studies on the physiological effects of non-polar-polar organic electrolytes; the influence of detergents upon the potentiometric reaction and the contractility of nerve and muscle

In a previous paper it has been shown that the nonpolar-polar anionic detergents can be divided into two main groups. One chemically characterized by a relatively long chain of non-polar alkyl groups, which in solution are in contact with one end of a muscle and, locally adhering to it, produce a permanent negative injury potential. This is generally accompanied by a loss of excitability. The second group, distinguished by a relatively short chain of non-polar alkyl groups acts reversibly, ordinarily preserves the excitability and, in contrast to the first group, produces locally a reverse positive potential. For reasons mentioned before, this appears likely to correspond to an increased activity. These concepts have been tested in this second paper. The measurements of the resting potentials of muscles have been supplemented by measurements on frog sciatics with the result that there are brought about, again by the detergents with long alkyl chains, regular irreversible negative resting potentials and with the short chain compounds reversible positive potentials are aroused. Furthermore, in addition there appeared the hardly expected result that muscle stimulated in the presence of short chain detergents responded with an even higher contraction. We have endeavored to explain this on the basis of general considerations concerning the physical chemistry of the excitatory process. More direct evidence of this rise of excitability under the influence of the short chain non-polar-polar detergents will be presented in the next papers on studies concerning chronaximetric measurements on nerve, referring particularly to the semidetergents, and concerning the effects of detergents in general upon the heart beat of a clam.     

Postsynaptic neuronal response of a cat sensorimotor cortex during evoked and self-sustained rhythmic "spike and wave" activity

Intracellular correlates of complex sets of rhythmic cortical "spike and wave" potentials evoked in sensorimotor cortex and of self-sustained rhythmic "spike and wave" activity were examined during acute experiments on cats immobilized by myorelaxants. Rhythmic spike-wave activity was produced by stimulating the thalamic relay (ventroposterolateral) nucleus (VPLN) at the rate of 3 Hz; self-sustained afterdischarges were recorded following 8–14 Hz stimulation of the same nucleus. Components of the spike and wave afterdischarge mainly correspond to the paroxysmal depolarizing shifts of the membrane potential of cortical neurons in length. After cessation of self-sustained spike and wave activity, prolonged hyperpolarization accompanied by inhibition of spike discharges and subsequent reinstatement of background activity was observed in cortical neurons. It is postulated that the negative slow wave of induced spike and wave activity as well as slow negative potentials of direct cortical and primary response reflect IPSP in more deep-lying areas of the cell bodies, while the wave of self-sustained rhythmic activity is due to paroxysmal depolarizing shifts in the membrane potential of cortical neurons.I. S. Beritashvili Institute of Physiology, Academy of Sciences of the Georgian SSR, Tbilisi. Translated from Neirofiziologiya, Vol. 18, No. 3, pp. 298–306, May–June, 1986.     

Effect of physical weathering on the carbon sequestration potential of biochars and hydrochars in soil

Physical weathering can modify the stability of biochar after field exposure. The aim of our study was to determine the potential carbon sequestration of the two chars at different timescales. We investigated the modification in composition and stability resulting from physical weathering of two different chars produced (i) at low temperature (250 °C) by hydrothermal carbonization (HTC); and (ii) at high temperature (1200 °C) by gasification (GS) using contrasting feedstocks. Physical weathering of HTC and GS placed on a water permeable canvas was performed through successive wetting/drying and freezing/thawing cycles. Carbon loss was assessed by mass balance. Chemical stability of the remaining material was evaluated as resistance to acid dichromate oxidation, and biological stability was assessed during laboratory incubation. Moreover, we assessed modification in potential priming effects due to physical weathering. Physical weathering induced a carbon loss ranging between 10 and 40% of the total C mass depending on the feedstock. This C loss is most probably related to leaching of small particulate and dissolved compounds. GS produced from maize silage showed the highest C loss. The chemical stability of HTC and GS was unaffected by physical weathering. In contrast, physical weathering strongly increased the biological stability of HTC and GS char produced from maize silage. After physical weathering, the half‐life (t_1/2) of GS was doubled but only slight increase was noted for those of HTC. During the first weeks of incubation, HTC addition to soil stimulated native soil organic matter (SOM) mineralization (positive priming effect), while the GS addition led to protection of the native SOM against biologic degradation (negative priming effect). Physical weathering led to reduction in these priming effects. Model extrapolations based on our data showed that decadal C sequestration potential of GS and HTC is globally equivalent when all losses including those due to priming and physical weathering were taken into account. However, at century scale only GS may have the potential to increase soil C storage.     

Effect of electromagnetic stimulation with different waveforms on cultured chick tendon fibroblasts

An energy efficient electromagnetic stimulator device for fracture healing was compared to a commercially available device in stimulating cell growth in tissue cultures. The energy efficient device, which conserves energy by using a bidirectional time-dependent magnetic wave form, and the commercially available stimulator, which uses a unidirectionaltime-dependent magnetic wave form, were tested on chick tendon fibroblasts in primary culture. Comparing non-stimulated control and cells electromagnetically stimulated with unidirectional and bidirectional waveforms showed that at the growth phase between days 2 and 3, both electrical stimulation techniques increased cell division as measured by DNA synthesis. When cells were dividing rapidly, collagen synthesis was reduced. When the cells reached the confluence there was no difference among the groups (control, unidirectionally stimulated, and bidirectionally stimulated) in terms of number of cells or collagen produced. © 1994 Wiley-Liss, Inc.     

Suppression of Brown Spot Disease of Cultivated Chrysanthemum by Manipulating Phototropic Response of Conidium Germ Tubes of Septoria obesa

An antagonistic effect between the UV-blue (shorter than 500 nm) and red (longer than 600 nm) wave regions on the induction of phototropic response in conidium germ tubes has been demonstrated in the fungus Septoria obesa, the causal organism of brown spot disease of cultivated chrysanthemum. Phototropic response of conidium germ tubes was dependent on the ratio of the fluence rates between negative phototropism-inducing UV-blue wave region and positive phototropism-inducing red wave region. Elimination of the UV wave region shorter than 390 nm by an ultraviolet absorbing vinyl film significantly reduced the negative phototropic response of conidium germ tubes and suppressed the invasion of the fungus through the stomata. Thus, a manipulation of the phototropic response of conidium germ tubes by controlling light quality resulted in the suppression of brown spot disease of cultivated chrysanthemum.