bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of sulfhydryl inhibitors on nonlinear membrane currents in frog skeletal muscle fibers.
Authors: Gonzalez A , Bolaños P , Caputo C
Published: May 2004

The effect of sulhydryl reagents on nonlinear membrane currents of frog skeletal muscle fibers has been studied using the triple Vaseline gap voltage-clamp technique. These compounds, which are known to interfere with depolarization contraction coupling, also appear to diminish intramembranous charge movement recorded with fibers polarized to -100 mV (charge 1). This effect, however, is accompanied by changes in the fiber membrane conductance and in most cases by the appearance of an inwardly directed current in the potential range between -60 and +20 mV. This current is reduced by both cadmium and nifedipine and does not occur in Ca-free solution, suggesting that it is carried by calcium ions flowing through regular calcium channels that are more easily activated in the presence of SH reagent. These changes in the membrane electrical active and passive properties decrease the quality and reliability of the P/n pulse subtracting procedure normally used for charge movement measurements. These effects can be substantially reduced by cadmium ions (0.1 mM), which has no effect on charge movement. When SH reagents are applied in the presence of cadmium, no effects are observed, indicating that this cation may protect the membrane from the reagent effects. The effects of -SH reagents can be observed by applying them in the absence of cadmium, followed by addition of the cation. Under these conditions the conductance changes are reversed and the effects of the SH reagents on charge movement can be measured with a higher degree of confidence. Maximum charge is reduced by 32% in the presence of 1.5 mM PCMB and by 31% in the presence of 2 mM PHMPS. These effects do not occur in the presence of DTT and in some cases they may be reversed by this agent. Charge 2, recorded in depolarized muscle fibers, is also reduced by these agents.
Functional role of the NH2-terminal cytoplasmic domain of a mammalian A-type K channel.
Authors: Tseng-Crank J , Yao J A , Berman M F , Tseng G N
Published: May 2004

It has been shown for a Shaker channel (H-4) that its NH2-terminal cytoplasmic domain may form a "ball and chain" structure, with the "chain" tethering the "ball" to the channel while the "ball" capable of binding to the channel in its open state and causing inactivation. Equivalent structures have not been identified in mammalian Shaker homologues. We studied the functional role of the NH2-terminal region of a fast-inactivating mammalian K channel, RHK1 (Kv1.4), by deleting different domains in this region and examining the resultant changes in channel properties at whole cell and single channel levels. Deleting the NH2-terminal hydrophobic domain (domain A) or the subsequent positive charges (domain I) from RHK1 greatly slowed the decay of whole cell currents, suggesting the existence of a ball-like structure in RHK1 similar to that in the Shaker channel. The function of the ball appeared to be abolished by deleting domain A, while modified but maintained by deleting domain I. In the latter case, the data suggest that the positive charges needed for the function of the ball can be replaced by amino acids from a following region (domain III) that has a high positive charge density. Deleting multiple domains from the NH2 terminus of RHK1 corresponding to the chain in Shaker H-4 did not induce expected changes in channel properties that might result from a shortening of a chain. A comparison of single channel kinetics of selected mutant channels with those of the wild-type channel indicated that these deletion mutations slowed whole cell currents by prolonging burst durations and by increasing the probability of reopening during depolarization. There were no changes in single channel current amplitude or latency to first opening. In conclusion, our observations indicate that the inactivation mechanism of RHK1 is similar to that of Shaker H-4 in that a positively charged cytoplasmic domain is important for such a process. The NH2-terminal domain is not involved in channel activation or ion permeation process.
Hyperpolarization-activated chloride currents in Xenopus oocytes.
Authors: Kowdley G C , Ackerman S J , John J E , Jones L R , Moorman J R
Published: May 2004

During hyperpolarizing pulses, defolliculated Xenopus oocytes have time- and voltage-dependent inward chloride currents. The currents vary greatly in amplitude from batch to batch; activate slowly and, in general, do not decay; have a selectivity sequence of I- > NO3- > Br- > Cl- > propionate > acetate; are insensitive to Ca2+ and pH; are blocked by Ba2+ and some chloride channel blockers; and have a gating valence of approximately 1.3 charges. In contrast to hyperpolarization-activated chloride currents induced after expression of phospholemman (Palmer, C. J., B. T. Scott, and L. R. Jones. 1991. Journal of Biological Chemistry. 266:11126; Moorman, J. R., C. J. Palmer, J. E. John, J. E. Durieux, and L. R. Jones. 1992. 267:14551), these endogenous currents are smaller; have a different pharmacologic profile; have a lower threshold for activation and lower voltage-sensitivity of activation; have different activation kinetics; and are insensitive to pH. Nonetheless, the endogenous and expressed current share striking similarities. Recordings of macroscopic oocyte currents may be inadequate to determine whether phospholemman is itself an ion channel and not a channel-modulating molecule.
Surface charge potentiates conduction through the cardiac ryanodine receptor channel.
Authors: Tu Q , Velez P , Cortes-Gutierrez M , Fill M
Published: May 2004

Single channel currents through cardiac sarcoplasmic reticulum (SR) Ca2+ release channels were measured in very low levels of current carrier (e.g., 1 mM Ba2+). The hypothesis that surface charge contributes to these anomalously large single channel currents was tested by changing ionic strength and surface charge density. Channel identity and sidedness was pharmacologically determined. At low ionic strength (20 mM Cs+), Cs+ conduction in the lumen-->myoplasm (L-->M) direction was significantly greater than in the reverse direction (301.7 +/- 92.5 vs 59.8 +/- 38 pS, P < 0.001; mean +/- SD, t test). The Cs+ concentration at which conduction reached half saturation was asymmetric (32 vs 222 mM) and voltage independent. At high ionic strength (400 mM Cs+), conduction in both direction saturated at 550 +/- 32 pS. Further, neutralization of carboxyl groups on the lumenal side of the channel significantly reduced conduction (333.0 +/- 22.5 vs 216.2 +/- 24.4 pS, P < 0.002). These results indicate that negative surface charge exists near the lumenal mouth of the channel but outside the electric field of the membrane. In vivo, this surface charge may potentiate conduction by increasing the local Ca2+ concentration and thus act as a preselection filter for this poorly selective channel.
Phosphate from the phosphointermediate (EP) of the human red blood cell Na/K pump is coeffluxed with Na, in the absence of external K.
Authors: Marín R , Hoffman J F
Published: May 2004

This study is concerned with Na/K pump-mediated phosphate efflux that occurs during uncoupled Na efflux in human red blood cells. Uncoupled Na efflux is known to be a ouabain-sensitive mode of the Na/K pump that occurs in the absence of external Nao and Ko. Because this efflux (measured with 22Na) is also inhibited by 5 mM Nao, the efflux can be separated into a Nao-sensitive and a Nao-insensitive component. Previous work established that the Nao-sensitive efflux is actually comprised of an electroneutral coefflux of Na with cellular anions, such as SO4 (as 35SO4). The present work focuses on the Nao-insensitive component in which the principal finding is that orthophosphate (P(i)) is coeffluxed with Na in a ouabain-sensitive manner. This P(i) efflux can be seen to occur, in the absence of Ko, in both DIDS-treated intact cells and resealed red cell ghosts. This efflux of P(i) was shown to be derived directly from the pump's substrate, ATP, by the use of resealed ghosts made to contain both ATP and P(i) in which either the ATP or the P(i) were labeled with, respectively, [gamma-32P]ATP or [32P]H3PO4. (These resealed ghosts also contained Na, Mg, P(i), SO4, Ap5A, as well as an arginine kinase/creatine kinase nucleotide regenerating system for the control of ATP and ADP concentrations, and were suspended usually in (NMG)2SO4 at pH 7.4.) It was found that 32P was only coeffluxed with Na when the 32P was contained in [gamma-32P]ATP and not in [32P]H3PO4. This result implies that the 32P that is released comes from ATP via the pump's phosphointermediate (EP) without commingling with the cellular pool of P(i). Ko (as K2SO4) inhibits this 32P efflux as well as the Nao-sensitive 35SO4 efflux, with a K0.5 of 0.3-0.4 mM. The K0.5 for inhibition of P(i) efflux by Ko is not influenced by Nao, nor can Nao act as a congenor for Ko in any of the flux reactions involving Ko. The stoichiometry of Na to SO4 and Na to P(i) efflux is approximately 2:1 under circumstances where the stoichiometry of Na effluxed to ATP utilized is 3:1. From these and other results reported, it is suggested that there are two types of uncoupled Na efflux that differ from each other on the basis of their sensitivity to Nao, the source (cellular vs substrate) and kind of anion (SO4 vs P(i)) transported.(ABSTRACT TRUNCATED AT 400 WORDS)
ADP+orthophosphate (P(i)) stimulates an Na/K pump-mediated coefflux of P(i) and Na in human red blood cell ghosts.
Authors: Marín R , Hoffman J F
Published: May 2004

The Na/K pump in human red blood cells that normally exchanges 3 Nai for 2 Ko is known to continue to transport Na in a ouabain-sensitive and ATP-dependent manner when the medium is made free of both Nao and Ko. Although this Na efflux is called "uncoupled" because of removal of ions to exchange with, the efflux has been shown to be comprised of a coefflux with cellular anions. The work described in this paper presents a new mode of operation of uncoupled Na efflux. This new mode not only depends upon the combined presence of ADP and intracellular orthophosphate (P(i))i but the Na efflux that is stimulated to occur is coeffluxed with (P(i))i. These studies were carried out with DIDS-treated resealed red cell ghosts, suspended in buffered (NMG)2SO4, that were made to contain, in addition to other constituents, varying concentrations of ADP and P(i) together with Na2 SO4, MgSO4 and hexokinase. While neither ADP nor P(i) was effective alone, ouabain-sensitive uncoupled Na efflux, (measured with 22Na) could be activated by [ADP+P(i)] where the K0.5 for ADP in the presence of 10 mmol (P(i))i/liter ghosts was 100-200 mumol/liter ghosts and the K0.5 for (P(i))i, in the presence of 500 mumol ADP/liter ghosts was 3-4 mmol/liter ghosts. [ADP+P(i)] activation of this Na efflux could be inhibited by as little as 2 mumol ATP/liter ghosts but the inhibition could be relieved by the addition of 50 mM glucose, given entrapped hexokinase. While ouabain-sensitive Na efflux was found to be coeffluxed with P(i) (measured with entrapped [32P]H3PO4), this was not so for SO4 (measured with 35SO4). The stoichiometry of Na to P(i) efflux was found to be approximately 2 to 1. Na efflux as well as (P(i))i efflux were both inhibited by 10 mM Nao (K0.5 approximately equal to 4 mM). But, whereas 20 mM Ko (K0.5 approximately equal to 6 mM) inhibited the efflux of (P(i))i, as would be expected from previous work, Na efflux was actually increased. When Ko influx was measured in this situation there was a 1 for 1 exchange of Nai for Ko, that is, of course, downhill with respect to the gradient of each ion. Surprisingly AsO4 was unable to replace P(i) for activation of Na efflux but Na efflux could be inhibited by vanadate and oligomycin. In terms of mechanism, it is likely that ADP acts to promote the formation of the phosphoenzyme (EP) by (P(i))i that would otherwise be inhibited by Nai.(ABSTRACT TRUNCATED AT 400 WORDS)
Modulation of synaptic transfer between retinal cones and horizontal cells by spatial contrast.
Authors: Cadenas I D , Reifsnider E S , Tranchina D
Published: May 2004

We studied the influence of steady annular light on the kinetics and sensitivity of horizontal cell (HC) responses to modulation of the intensity of small concentric spots in the turtle retina. As shown by previous investigators, when the intensity of the annulus was equal to the mean spot intensity, spot response kinetics were the same as those for the modulation of spatially uniform light. Turning off the annulus attenuated dramatically high-frequency flicker sensitivity and enhanced somewhat low-frequency sensitivity. This phenomenon reflects a modulation of synaptic transfer between cones and second-order neurons that is mediated by cones, and it will be referred to as cone-mediated surround enhancement (CMSE). Our main results are as follows: (a) The change in test-spot response sensitivity and kinetics upon dimming a steady surrounding annulus is a consequence of the change in spatial contrast rather than change in overall light level. (b) Introduction of moderate contrast between the mean spot intensity and steady surrounding light intensity causes a marked change in spot response kinetics. (c) The dependence of spot response kinetics on surrounding light can be described by a phenomenological model in which the steady state gain and the time constant of one or two single-stage, low-pass filters increase with decreasing annular light intensity (d) The effect of surrounding light on spot responses of a given HC is not determined by change in the steady component of the membrane potential of that cell. (e) Light outside the receptive field of an HC can affect that cell's spot response kinetics. (f) In an expanding annulus experiment, the distance over which steady annular light affects spot response kinetics varies among HCs and can be quite different even between two cells with closely matched receptive field sizes. (g) The degree of CMSE is correlated with HC receptive field size. This correlation suggests that part of the enhancement mechanism is located in the HC. Taken together, our results suggest the involvement of the inner retina in CMSE.
Evaluation of the rate constants of sugar transport through maltoporin (LamB) of Escherichia coli from the sugar-induced current noise.
Authors: Andersen C , Jordy M , Benz R
Published: May 2004

LamB (maltoporin) of Escherichia coli outer membrane was reconstituted into artificial lipid bilayer membranes. The channel contains a binding site for sugars and is blocked for ions when the site is occupied by a sugar. The on and off reactions of sugar binding cause an increase of the noise of the current through the channel. The sugar-induced current noise of maltoporin was used for the evaluation of the sugar-binding kinetics for different sugars of the maltooligosaccharide series and for sucrose. The on rate constant for sugar binding was between 10(6) and 10(7) M-1.s-1 for the maltooligosaccharides and corresponds to the movement of the sugars from the aqueous phase to the central binding site. The off rate (corresponding to the release of the sugars from the channel) decreased with increasing number of glucose residues in the maltooligosaccharides from approximately 2,000 s-1 for maltotriose to 180 s-1 for maltoheptaose. The kinetics for sucrose movement was considerably slower. The activation energies of the stability constant and of the rate constants for sugar binding were evaluated from noise experiments at different temperatures. The role of LamB in the transport of maltooligosaccharides across the outer membrane is discussed.
Characterization of the voltage-dependent properties of a volume-sensitive anion conductance.
Authors: Jackson P S , Strange K
Published: May 2004

Outwardly rectified, swelling-activated anion conductances have been described in numerous cell types. The major functional variable observed amongst these conductances is the extent and rate of depolarization-induced inactivation. In general, the conductances can be divided into two broad classes, those that show rapid inactivation in response to strong depolarization and those that show little or no voltage dependence. The swelling-activated anion conductance in rat C6 glioma cells is inactivated nearly completely by membrane depolarization above +90 mV and reactivated by membrane hyperpolarization. The kinetics of inactivation and reactivation are fit by single and double exponentials, respectively. Voltage-dependent behavior is well described by a simple linear kinetic model in which the channel exists in an open or one of three inactivated states. pH-induced changes in voltage-dependent gating suggest that the voltage sensor contains critical basic amino acid residues. Extracellular ATP blocks the channel in a voltage-dependent manner. The block is sensitive to the direction of net Cl- movement and increases open channel noise indicating that ATP interacts with the channel pore. Blockage of the channel with ATP dramatically slows depolarization-induced inactivation.
Cd2+ regulation of the hyperpolarization-activated current IAB in crayfish muscle.
Authors: Araque A , Cattaert D , Buño W
Published: May 2004

The effects of Cd2+ on the hyperpolarization-activated K(+)-mediated current called IAB (Araque, A., and W. Buño. 1994. Journal of Neuroscience. 14:399-408.) were studied under two-electrode voltage-clamp in opener muscle fibers of the crayfish Procambarus clarkii. IAB was reversibly reduced by extracellular Cd2+ in a concentration-dependent manner, obeying the Hill equation with IC50 = 0.452 +/- 0.045 mM and a Hill coefficient of 1 (determined from the maximal chord conductance of IAB). Cd2+ decreased the IAB conductance (GAB) and shifted its voltage dependence towards hyperpolarized potentials in a similar degree, without affecting the slope of the voltage dependence. The IAB activation time constant increased, whereas the IAB deactivation time constant was not modified by Cd2+. The IAB equilibrium potential (EAB) was unmodified by Cd2+, indicating that the selective permeability of IAB channels was not altered. IAB was unaffected by intracellular Cd2+. The Cd(2+)-regulation of IAB did not depend on [K+]o, and the effects of [K+]o on IAB were unchanged by Cd2+, indicating that Cd2+ did not compete with K+. Therefore, Cd2+ probably bound to a different site to that involved in the K+ permeability pathway. We conclude that Cd2+ affected the gating of IAB channels, interfering with their opening but not with their closing mechanism. The results can be explained by a kinetic model in which the binding of Cd2+ to the IAB channels would stabilize the gating apparatus at its resting position, increasing the energy barrier for the transition from the closed to the open channel states.
The kinetics of inactivation of the rod phototransduction cascade with constant Ca2+i.
Authors: Lyubarsky A , Nikonov S , Pugh E N
Published: May 2004

A rich variety of mechanisms govern the inactivation of the rod phototransduction cascade. These include rhodopsin phosphorylation and subsequent binding of arrestin; modulation of rhodopsin kinase by S-modulin (recoverin); regulation of G-protein and phosphodiesterase inactivation by GTPase-activating factors; and modulation of guanylyl cyclase by a high-affinity Ca(2+)-binding protein. The dependence of several of the inactivation mechanisms on Ca2+i makes it difficult to assess the contributions of these mechanisms to the recovery kinetics in situ, where Ca2+i is dynamically modulated during the photoresponse. We recorded the circulating currents of salamander rods, the inner segments of which are held in suction electrodes in Ringer's solution. We characterized the response kinetics to flashes under two conditions: when the outer segments are in Ringer's solution, and when they are in low-Ca2+ choline solutions, which we show clamp Ca2+i very near its resting level. At T = 20-22 degrees C, the recovery phases of responses to saturating flashes producing 10(2.5)-10(4.5) photoisomerizations under both conditions are characterized by a dominant time constant, tau c = 2.4 +/- 0.4 s, the value of which is not dependent on the solution bathing the outer segment and therefore not dependent on Ca2+i. We extended a successful model of activation by incorporating into it a first-order inactivation of R*, and a first-order, simultaneous inactivation of G-protein (G*) and phosphodiesterase (PDE*). We demonstrated that the inactivation kinetics of families of responses obtained with Ca2+i clamped to rest are well characterized by this model, having one of the two inactivation time constants (tau r* or tau PDE*) equal to tau c, and the other time constant equal to 0.4 +/- 0.06 s.
A slow component of intramembranous charge movement during sarcoplasmic reticulum calcium release in frog cut muscle fibers.
Authors: Pape P C , Jong D S , Chandler W K
Published: May 2004

Cut muscle fibers from Rana temporaria were mounted in a double Vaseline-gap chamber and equilibrated with an end-pool solution that contained 20 mM EGTA and 1.76 mM Ca (sarcomere length, 3.3-3.8 microns; temperature, 14-16 degrees C). Sarcoplasmic reticulum (SR) Ca release, delta[CaT], was estimated from changes in myoplasmic pH (Pape, P.C., D.-S. Jong, and W.K. Chandler. 1995. J. Gen. Physiol. 106:259-336). The maximal value of delta[CaT] obtained during a depleting depolarization was assumed to equal the SR Ca content before stimulation, [CaSR]R (expressed as myoplasmic concentration). After a depolarization to -55 to -40 mV in fibers with [CaSR]R = 1,000-3,000 microM, currents from intramembranous charge movement, Icm, showed an early I beta component. This was followed by an I gamma hump, which decayed within 50 ms to a small current that was maintained for as long as 500 ms. This slow current was probably a component of Icm because the amount of OFF charge, measured after depolarizations of different durations, increased according to the amount of ON charge. Icm was also measured after the SR had been depleted of most of its Ca, either by a depleting conditioning depolarization or by Ca removal from the end pools followed by a series of depleting depolarizations. The early I beta component was essentially unchanged by Ca depletion, the I gamma hump was increased (for [CaSR]R > 200 microM), the slow component was eliminated, and the total amount of OFF charge was essentially unchanged. These results suggest that the slow component of ON Icm is not movement of a new species of charge but is probably movement of Q gamma that is slowed by SR Ca release or some associated event such as the accompanying increase in myoplasmic free [Ca] that is expected to occur near the Ca release sites. The peak value of the apparent rate constant associated with this current, 2-4%/ms at pulse potentials between -48 and -40 mV, is decreased by half when [CaSR]R approximately equal to 500-1,000 microM, which gives a peak rate of SR Ca release of approximately 5-10 microM/ms.
Ca2+ fluxes and channel regulation in rods of the albino rat.
Authors: Knopp A , Rüppel H
Published: May 2004

By use of microelectrodes, changes in the receptor current and the Ca2+ concentration were measured in the rod layer of the rat retina after stimulation by flashes or steady light. Thereby light induced Ca2+ sources, and sinks along a rod were determined in dependence of time. Thus, the Ca2+ fluxes across the plasma membrane of a mammalian rod could be studied in detail. By light stimulation, Ca2+ sources are evoked along the outer segment only. Immediately after a saturating flash, a maximum of Ca2+ efflux is observed which decays exponentially with tau = 0.3 s at 37 degrees C (4.2 s at 23 degrees C). During regeneration of the dark current, the outer segment acts as a Ca2+ sink, indicating a restoration of the Ca(2+)-depleted outer segment. These findings agree with earlier reports on amphibian rods. Further experiments showed that the peak Ca2+ efflux and tau are temperature dependent. The peak amplitude also depends on the external Ca2+ concentration. In contrast to the reports on amphibian rods, only a part of the Ca2+ ions extruded from the outer segment is directly restored. Surprisingly, during steady light the Ca2+ efflux approaches a permanent residual value. Therefore, in course of a photoresponse, Ca2+ must be liberated irreversibly from internal Ca2+ stores. There is certain evidence that the inner segment acts as a Ca2+ store. Our results show that the Ca2+ fraction of the ions carrying the dark current is proportional to the extracellular Ca2+ concentration. This indicates that the Ca2+ permeability of the plasma membrane of the rod outer segment is independent of the Ca2+ concentration.
Hypothesis for a serine proteinase-like domain at the COOH terminus of Slowpoke calcium-activated potassium channels.
Authors: Moss G W , Marshall J , Moczydlowski E
Published: May 2004

Bovine pancreatic trypsin inhibitor (BPTI) is a 58-residue protein with three disulfide bonds that belongs to the Kunitz family of serine proteinase inhibitors. BPTI is an extremely potent inhibitor of trypsin, but it also specifically binds to various active and inactive serine proteinase homologs with KD values that range over eight orders of magnitude. We previously described an interaction of BPTI at an intracellular site that results in the production of discrete subconductance events in large conductance Ca2+ activated K+ channels (Moss, G.W.J., and E. Moczydlowski. 1996, J. Gen. Physiol, 107:47-68). In this paper, we summarize a variety of accumulated evidence which suggests that BPTI binds to a site on the KCa channel protein that structurally resembles a serine proteinase. One line of evidence includes the finding that the complex of BPTI and trypsin, in which the inhibitory loop of BPTI is masked by interaction with trypsin, is completely ineffective in the production of substate events in the KCa channel. To further investigate this notion, we performed a sequence analysis of the alpha-subunit of cloned slowpoke KCa channels from Drosophila and mammals. This analysis suggests that a region of approximately 250 residues near the COOH terminus of the KCa channel is homologous to members of the serine proteinase family, but is catalytically inactive because of various substitutions of key catalytic residues. The sequence analysis also predicts the location of a Ca(2+)-binding loop that is found in many serine proteinase enzymes. We hypothesize that this COOH-terminal domain of the slowpoke KCa channel adopts the characteristic double-barrel fold of serine proteinases, is involved in Ca(2+)-activation of the channel, and may also bind other intracellular components that regulate KCa channel activity.
MICRURGICAL STUDIES IN CELL PHYSIOLOGY
Authors: Reznikoff Paul
Published: May 2004

I. The Plasmalemma.

1. On the plasmalemma of amebæ CaCl2 antagonizes the toxic action of LiCl better than it does NaCl, and still better than it does KCl. MgCl2 antagonizes the toxic action of NaCl better than it does LiCl and still better than it does KCl.

2. CaCl2 antagonizes the toxic action of LiCl and of KCl better than does MgCl2: MgCl2 antagonizes NaCl better than does CaCl2.

II. The Internal Protoplasm.

3. The antagonizing efficiency of CaCl2 and of MgCl2 are highest against the toxic action of KCl on the internal protoplasm, less against that of NaCl, and least against that of LiCl.

4. CaCl2 antagonizes the toxic action of LiCl better than does MgCl2: MgCl2 antagonizes the toxic action of NaCl and of KCl better than does CaCl2.

5. LiCl antagonizes the toxic action of MgCl2 on the internal protoplasm more effectively than do NaCl or KCl, which have an equal antagonizing effect on the MgCl2 action.

III. The Nature of Antagonism.

6. When the concentration of an antagonizing salt is increased to a toxic value, it acts synergistically with a toxic salt.

7. No case was found in which a potentially antagonistic salt abolishes the toxic action of a salt unless it is present at the site (surface or interior) of toxic action.

8. Antagonistic actions of the salts used in these experiments are of differing effectiveness on the internal protoplasm and on the surface membrane.
THE EFFECT OF RENNIN UPON CASEIN
Authors: Pertzoff Vladimir
Published: May 2004

The properties of the paracasein and casein preparations studied are compared in Table VI.

See PDF for Structure

I. Casein retains its characteristic solubility in NaOH: (1) after being exposed to a high degree of alkalinity during its preparation, (2) when recovered from partially hydrolyzed solutions in NaOH, and (3) after being kept for a prolonged time at the isoelectric point at 5°C.

II. It follows from I, that: (1) paracasein is not identical to casein modified by an excess of alkali, and that (2) this protein was not produced from casein by a partial hydrolysis of the latter in presence of NaOH.
ON THE PLACE OF PHOTIC ADAPTATION
Authors: Crozier W. J. , Wolf E.
Published: May 2004

The progress of photic adaptation of Agriolimax, when studied by the method of compounding phototropic and geotropic vectors, is shown to be uninfluenced by the concurrent gravitational excitation. Direct proof is thus obtained that the adaptation to light, manifest in its steadily decreasing effectiveness as a stimulus during the course of exposure, is not due to any central nervous adjustment simulating "learning," but is due to photochemical changes in the receptors.
TEMPERATURE CHARACTERISTICS FOR FREQUENCY OF RESPIRATORY MOVEMENTS IN YOUNG MAMMALS
Authors: Stier T. J. B. , Pincus G.
Published: May 2004

The internal temperature of 2 day old mice deviates by +0.01° to +3.0°C. from the environmental temperature over a range of 24°. The undeveloped temperature control allows rhythmic activities in this mammalian material to be readily subjected to the analysis afforded by temperature characteristics, and thus makes possible some insight into the physicochemical events controlling vital processes in entire uninjured mammals. The thermal increments and critical temperatures obtained point to a similarity in the controlling system of reactions for both homothermic and poikilothermic organisms. For frequency of respiratory movements the increments 12,340 (19.6° to 35.2°C.); 28,340 or 36,500 (15.5° to 19.6°C.) are most frequently found (thirteen cases). Rarely µ = 8,450, 33,000, or 18,340 (two cases). The last increment is either associated with µ = 36,000 below 20°, or extends unbroken throughout the whole range (one case).
THE NATURE OF THE IONIZABLE GROUPS IN PROTEINS
Authors: Simms Henry S.
Published: May 2004

Analysis of the experimental titration curves shows that gelatin contains acid groups with dissociation indices at pH 2.9 to 3.5 corresponding quantitatively with the content in dicarboxylic amino acids; and that the acidic group at pH 9.4 in egg albumin agrees with the amount of tyrosine.

The amounts of histidine and lysine present in both these proteins agree quantitatively with basic groups at pH 6.1 and pH 10.4 to 10.6, respectively.

However, the quantity of the arginine group (pH 8.1) in these proteins is considerably less than the amount of arginine found on hydrolysis. This deficiency is compensated (quantitatively with gelatin and approximately with egg albumin) by a basic group at pH 4.6.

The structure of this "4.6 group" should be similar to aniline and cytosine in consisting of an amino group on a conjugated unsaturated (perhaps cyclic) system. It would appear that the 4.6 group is disrupted on hydrolysis, producing arginine, and may be referred to as "prearginine."

The presence of prearginine in proteins, instead of the full amount of arginine, has an important effect on the properties. Otherwise the isoelectric point of gelatin would be 8.0 (instead of 4.7) and of egg albumin 6.6 (instead of 4.8), and the titration curves would be quite different in shape between pH 4 and 10.

Deamination of gelatin produces no decrease in prearginine, arginine, or histidine groups, but removes nearly all of the lysine group.
THE INFLUENCE OF TEMPERATURE ON THE PHOTOSENSORY LATENT PERIOD
Authors: Hecht Selig
Published: May 2004

1. The effect of temperature on the photosensory latent period in Pholas dactylus is accurately described by the Arrhenius equation when µ = 18,300.

2. The adequacy of this equation has already been found for two other photosensitive animals, Mya and Ciona, which are very similar in behavior to Pholas. The value of µ is different for each of the three species studied.

3. This is taken to mean that though the organization of the receptor process is the same for the three species, the chemical materials concerned are very likely different.
SIGNIFICANCE OF THE CHEMICAL COMPOSITION OF THE SECRETING AND DRY MAMMARY GLAND TO MILK SECRETION
Authors: Gowen John W. , Tobey Elmer R.
Published: May 2004

The results herein presented furnish exact critical evidence for one more stage in milk secretion. Cows producing up to 30 pounds of milk at one milking are shown to have the lactose equivalent of all this milk in the udder when milking commences. The average excess of lactose found in the udder after subtracting the amount necessary for the contained milk is equal to 2.1 pounds. This represents the milk retained in the udder when the cow is believed to be dry. These conclusions are further supported by the fact that no sugar is found in the udder in the quiescent state.

The study of the total composition of the udder as fat, ash, nitrogen, and lactose, and of the contained milk shows that there is a large excess of fat, ash, and nitrogen in proportion to that necessary for milk formation. The excess of udder lactose over the milk lactose is much less.

The lactose would therefore appear to be formed from some element in the blood, probably dextrose, only as needed for the formation of milk.

The composition of the dry udder is quite different in certain respects from that of the actively secreting gland. It builds up a fat reserve of a quite different Reichert-Meissl number from that of butter-fat. It has no sugar, its ash content is reduced, and the nitrogen content is like that of the secreting gland.
THE PREARGININE IN EDESTIN AND ITS RESISTANCE TO HYDROLYSIS
Authors: Simms Henry S.
Published: May 2004

The titration data of edestin show that all the arginine found on hydrolysis exists in this protein as "prearginine."

The extra ionizable groups of histidine, lysine and tyrosine are free in the quantities found on hydrolysis. Part of the extra carboxyl groups of aspartic and glutamic acids are bound as amides, and 50 per cent are bound in some other manner (perhaps anhydride) leaving only about 6 per cent of these groups free to ionize in edestin.

The prearginine in edestin is not converted into arginine on hydrolysis with pepsin up to 18 per cent (of the total hydrolysis). In more highly hydrolyzed solutions it is not possible to detect such a conversion, due to high buffering. Complete hydrolysis however converts prearginine into arginine which can be isolated.

Hydrolyzed edestin promotes the growth of sarcomatous fibroblasts about equally well whether 5, 14 or 18 per cent hydrolyzed.
SYNERESIS AND SWELLING OF GELATIN
Authors: Kunitz M.
Published: May 2004

1. When solid blocks of isoelectric gelatin are placed in cold distilled water or dilute buffer of pH 4.7, only those of a gelatin content of more than 10 per cent swell, while those of a lower gelatin content not only do not swell but actually lose water.

2. The final quantity of water lost by blocks of dilute gelatin is the same whether the block is immersed in a large volume of water or whether syneresis has been initiated in the gel through mechanical forces such as shaking, pressure, etc., even in the absence of any outside liquid, thus showing that syneresis is identical with the process of negative swelling of dilute gels when placed in cold water, and may be used as a convenient term for it.

3. Acid- or alkali-containing gels give rise to greater syneresis than isoelectric gels, after the acid or alkali has been removed by dialysis.

4. Salt-containing gels show greater syneresis than salt-free gels of the same pH, after the salt has been washed away.

5. The acid and alkali and also the salt effect on syneresis of gels disappears at a gelatin concentration above 8 per cent.

6. The striking similarity in the behavior of gels with respect to syneresis and of gelatin solutions with respect to viscosity suggests the probability that both are due to the same mechanism, namely the mechanism of hydration of the micellæ in gelatin by means of osmosis as brought about either by diffusible ions, as in the presence of acid or alkali, or by the soluble gelatin present in the micellæ. The greater the pressures that caused swelling of the micellæ while the gelatin was in the sol state, the greater is the loss of water from the gels when the pressures are removed.

7. A quantitative study of the loss of water by dilute gels of various gelatin content shows that the same laws which have been found by Northrop to hold for the swelling of gels of high concentrations apply also to the process of losing water by dilute gels, i.e. to the process of syneresis. The general behavior is well represented by the equations:

See PDF for Equation

and

See PDF for Equation

where P1 = osmotic pressure of the soluble gelatin in the gel, P2 = stress on the micellæ in the gelatin solution before setting, Ke = bulk modulus of elasticity, Vo = volume of water per gram of dry gelatin at setting and Ve = volume of water per gram of gelatin at equilibrium.
MODIFICATION OF THE NORTHROP-KUNITZ MICROCATAPHORESIS CELL
Authors: Abramson Harold A.
Published: May 2004

A modification of the Northrop-Kunitz microcataphoresis cell is described. Although the theory of von Smoluchowski relating to such systems is in general followed, certain deviations may be encountered which are easily avoided.
THE PHOTOTROPIC EXCITATION OF LIMAX
Authors: Crozier W. J. , Cole W. H.
Published: May 2004

The photic orientation of Limax creeping geotropically upon a vertical plate is such that the phototropic vector determining the angular deflection ß from the vertical path is proportional to log I. This is proved by the fact that with horizontal illumination tan ß is directly proportional to log I; with non-horizontal light rays from a small source the ratio

See PDF for Equation

is directly proportional to log I (where A = the angle between light rays and the path of orientation), the vector diagram of the field of excitation being in this case not a right-angled triangle.
A GLASS ELECTRODE APPARATUS FOR MEASURING THE pH VALUES OF VERY SMALL VOLUMES OF SOLUTION
Authors: MacInnes D. A. , Dole Malcolm
Published: May 2004

A glass electrode apparatus is described with which pH measurements can be made with as small volumes as 2 drops (about 0.14 cc.) of solution.

Using this apparatus the change of pH of the vacuolar sap of Nitella, due to the penetration of brilliant cresyl blue, has been readily followed. The sap and the dye have been found to poison the usual type of hydrogen electrode.
RESEMBLANCES BETWEEN THE ELECTROMOTOR VARIATIONS OF RHYTHMICALLY REACTING LIVING AND NON-LIVING SYSTEMS
Authors: Lillie Ralph S.
Published: May 2004

1. The electromotor variations of pure iron wires, arranged to react rhythmically with nitric acid, are recorded and described.

2. Resemblances between these variations and those of rhythmically reacting living tissues (especially the heart) are pointed out and discussed.
THE CATALYTIC EFFECT OF DYES ON THE OXYGEN CONSUMPTION OF LIVING CELLS
Authors: Barron E. S. Guzman , Hoffman L. A.
Published: May 2004

From the experiments described in this paper and in those previously published it can be concluded that dyes which can be reversibly oxidized and reduced, act as catalysts for some oxidative processes taking place in the living cells, as is manifested by an increase in their oxygen consumption.

It has been found that the catalytic power of the dyes on the oxygen consumption of starfish eggs (mature, unfertilized) is conditioned by two factors: the reduction potential of the dye and the permeability of the cell surface. Dyes whose E'o is towards the positive side of the aerobic reduction potential of the starfish eggs have a maximum catalytic effect. This catalytic power decreases as the E'o becomes more negative than the reduction potential of the cell and becomes nil beyond certain limits. When a dye cannot penetrate into the cell, its effect is greatly diminished as in this case only those oxidative processes taking place at the outer surface of the cell can be activated.

Whether a dye can act as a catalyst or not is dependent on whether the normal consumption of oxygen by the cell is slower or quicker than the oxidation activated by the dye. The speed of this activation is correlated to (1) the speed at which the dye is reduced by the cell, and (2) the speed at which the leuco-dye is oxidized by the atmospheric oxygen. If one of these two processes is slower than the normal respiration, the dye cannot increase the rate of oxygen consumption (phenol indophenol at low concentrations which is kept reduced by the cell is very slowly reoxidized by atmospheric oxygen, on the other hand safranin and neutral red which are not reduced by the cell or at least too slowly reduced, though rapidly reoxidized by air). It will depend on these two reactions velocities whether a dye will act as catalyst (methylene blue and dyes with similar E'o which are quickly reduced by the cell and the leuco-dyes of which are relatively quickly reoxidized). Though this relationship between the reduction potential of the dyes and its catalytic power would seem in contradiction with the well known thermodynamic assumption that there is in general no distinct relationship between the potential and velocity of the reaction, we have pointed out from the literature some of the various experiments where one does recognize this connection.
THE INFLUENCE OF THE QUANTITY OF NUTRITION UPON THE GROWTH OF THE SUCKLING MOUSE
Authors: MacDowell E. C. , Gates W. H. , MacDowell C. G.
Published: May 2004

Under usual conditions, the growth of the suckling mouse is limited by the quantity of mother's milk available.

As this limitation is removed the growth curve approaches a parabola, which is abruptly interrupted at the end of the second week, when the natural process of weaning begins.
THE OSMOTIC PRESSURE OF CONCENTRATED SOLUTIONS OF GELATIN IN EQUILIBRIUM WITH SOLUTIONS OF MAGNESIUM CHLORIDE
Authors: Adair G. S. , Callow E. H.
Published: May 2004

The osmotic pressures and the membrane equilibrium of chloride ions have been determined for solutions of gelatin in equilibrium with solutions of magnesium chloride containing from 4.0 to 9.0 equivalents per litre.

The pressures increase more rapidly than the concentration, an effect represented by a high value of the term "b" in van der Waals' equation p (V – b) = RT. Calculations made by a thermodynamical formula which makes allowances for deviations from the ideal solution laws show that the high value of "b" is not due to an unequal distribution of diffusible ions.

The theory that the high values of the hydration estimated from viscosity formulae account for the magnitude of "b" has been examined and the conclusion has been reached that the term "b" for gelatin as well as for haemoglobin is considerably larger than the volume of the protein hydrate.
STUDIES ON PENETRATION OF DYES WITH GLASS ELECTRODE
Authors: Irwin Marian
Published: May 2004

Glass electrode measurements of the pH value of the sap of cells of Nitella show that azure B in the form of free base penetrates the vacuoles and raises the pH value of the sap to about the same degree as the free base of the dye added to the sap in vitro, but the dye salt dissolved in the sap does not alter the pH value of the sap. It is concluded that the dye penetrates the vacuoles chiefly in the form of free base and not as salt.

The dye from methylene blue solution containing azure B free base as impurity penetrates and accumulates in the vacuole. This dye must be azure B in the form of free base, since it raises the pH value of the sap to about the same extent as the free base of azure B dissolved in the sap in vitro. The dye absorbed by the chloroform from methylene blue solution behaves like the dye penetrating the vacuole. These results confirm those of spectrophotometric analysis previously published.

Crystal violet exists only in one form between pH 5 and pH 9.2, and does not alter the pH value of the sap at the concentrations used. It does not penetrate readily unless cells are injured.

A theory of "multiple partition coefficients" is described which explains the mechanism of the behavior of living cells to these dyes.

When the protoplasm is squeezed into the sap, the pH value of the mixture is higher than that of the pure sap. The behavior of such a mixture to the dye is very much like that of the sap except that with azure B and methylene blue the rise in the pH value of such a mixture is not so pronounced as with sap when the dye penetrates into the vacuoles.

Spectrophotometric measurements show that the dye which penetrates from methylene blue solution has a primary absorption maximum at 653 to 655 mµ (i.e., is a mixture of azure B and methylene blue, with preponderance of azure B) whether we take the sap alone or the sap plus protoplasm.

These results confirm those previously obtained with spectrophotometric measurements.
ON THE TEMPERATURE CHARACTERISTICS FOR FREQUENCY OF BREATHING MOVEMENTS IN INBRED STRAINS OF MICE AND IN THEIR HYBRID OFFSPRING. I
Authors: Pincus Gregory
Published: May 2004

Young mice of a selected line of the dilute brown strain of mice exhibit over the range 15–25°C. (body temperature) a relation of frequency of breathing movements to temperature such that when fitted by the Arrhenius equation the data give a value for the constant µ of 24,000± calories or, less frequently, 28,000±. Young mice of an inbred albino strain show over the range 15–20°C. a value of µ = 34,000±, or, less frequently, 14,000±, with a critical temperature at about 20°C. and a value of µ = 14,000± above 20°C. The F1 hybrids of these two strains, and the backcross generations to either parent strain, exhibit only those four values of the temperature characteristic observed in the parent strains and none other. One may therefore speak of the inheritance of the value of the constant µ, but the inheritance shows in this instance no Mendelian behavior. Furthermore there appears to be inherited the occurrence (or absence) of a critical temperature at 20°C. These experiments indicate the "biological reality" of the temperature characteristics.
TEMPERATURE CHARACTERISTICS FOR THE OXYGEN CONSUMPTION OF GERMINATING SEEDS OF LUPINUS ALBUS AND ZEA MAYS
Authors: Tang Pei-Sung
Published: May 2004

The rate of oxygen consumption by germinating seeds of Lupinus albus and of Zea mays was studied as a function of temperature (7–26°C.). The Warburg manometer technique was used, with slight modifications. Above and below a critical temperature at 19.5°C. the temperature characteristic for oxygen consumption by Lupinus albus was found to be µ = 11,700± and 16,600 respectively. The same critical temperature was encountered in the case of Zea mays, with temperature characteristics µ = 13,100± above and µ = 21,050 below that temperature.
STUDIES ON CRYSTALLINE UREASE
Authors: Tauber Henry , Kleiner Israel S.
Published: May 2004

1. Crystalline urease is not inactivated by trypsin in the absence of a gum. In fact, the presence of trypsin alone in aqueous solutions of urease has an action similar to that of gum, that is, it acts as a "protective colloid" for urease.

2. Crystalline urease is inactivated by trypsin in the presence of a gum. This occurs with great rapidity if purified (crystalline) trypsin is used.

3. If trypsin is added to urease a considerable time before the addition of gum no inactivation of the urease takes place.

4. The suggested explanation is that an inhibitive group of the urease molecule ("antitrypsin") may combine with trypsin to form an irreversible inactive trypsin compound, which cannot attack the urease but has unchanged ureolytic power. This compound cannot be formed if gum is present because the gum has united with a portion of the urease molecule; the tryptic power is thus unimpaired and urease is then digested. This speaks for the protein nature of urease.