Plasma kallikrein levels in acute Babesia argentina infections in splenectomised and intact calves.
Explore the source record for details and available documents.
Agar↗
SEARCH · Search PubMed
Results for “Tosyl Compounds”
Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
Chloramine-T effect on sodium conductance of neuroblastoma cells as studied by whole-cell clamp and single-channel analysis.
Patch-clamp experiments were done on sodium channels of neuroblastoma cells (N1E-115) in the presence of tetraethylammonium ions to block potassium channels. In Ringer solution whole-cell records revealed a diphasic INa inactivation with the fast (tau 0) component. being clearly larger than the slow (tau 1 approximately 3 tau 0) component. In single-channel studies on inside-out patches the mean open time, to, turned out to be only a fraction of tau 0 and almost independent of membrane potential. After external application of chloramine-T INa inactivation of whole cells was delayed with both tau 0 and tau 1 increased, and incomplete, i.e. a persistent current component emerged. The latter was maximal at a more positive membrane potential than the peak current. Also, after chloramine-T treatment the peak INa increased, particularly at weak depolarizations. In inside-out patches the equally effective internal application of chloramine-T led to bursting channel openings with mean burst times (tb) approximately 6 ms, and gap times (tg) approximately 20 ms, where gap is defined as a closure of greater than or equal to 1.5 ms. Within the bursts to was approximately 2 ms, again clearly shorter than tau 0; the mean close time, tc was approximately 0.5 ms. The single-channel conductance was approximately 13 pS and unaffected by chloramine-T. Diphasic INa inactivation and the fact that to less than tau 0 led to an extension of the model of Aldrich and Stevens [J Neurosci 7:418-431 (1987)], in which overall kinetics is determined by the openings rather than closures of the sodium channels.(ABSTRACT TRUNCATED AT 250 WORDS)
Chloramines↗
Interaction of 4-aminopyridine with normal and chloramine-T-modified K channels of neuroblastoma cells.
The steady-state effects and rate of action of 4-aminopyridine (4-AP) on normal and chloramine-T (CL-T)-modified voltage-dependent potassium (K) currents were studied in neuroblastoma cells with the whole-cell voltage-clamp current recording technique. 4-AP apparently slows both the activation and inactivation of the normal current but does not modify the time course of the CL-T-modified current. These differential effects of 4-AP are interpreted as resulting from the existence of two types of K channels with different 4-AP sensitivities under normal conditions and similar 4-AP sensitivities after CL-T, which furthermore slows their inactivation [8, 9]. While the onset of 4-AP action on the normal current is delayed and can be described by the difference of two exponentials, the onset of 4-AP action on CL-T-modified current starts immediately after the external application of the drug and can be described by the sum of two exponentials. The 4-AP-induced block of the normal current exhibits use-dependent features and is relieved by long conditioning depolarizations. In contrast, the block of the CL-T-modified current is not use-dependent. At high 4-AP concentrations (1-10 mM), the steady-state block of the normal current reaches a saturating value of 95%, while the steady-state block of the CL-T-modified current and the "unblocked" normal current only reaches a saturating value of 35%. The results suggest that CL-T inhibits a channel or membrane constituent which contributes to the inactivation of channels and increases their apparent affinity for 4-AP when they are in closed or open states.(ABSTRACT TRUNCATED AT 250 WORDS)
4-Aminopyridine↗
Atopic allergy to chloramine-T and the demonstration of specific IgE antibodies by the radioallergosorbent test.
Chloramine-T is a small molecular oxidizing agent that has been widely used as a disinfectant since the beginning of this century. It is generally used in a 5% solution but it is also supplied in powder form. Sporadic case reports of immediate-type sensitization to this agent associated with symptoms of asthma, rhinitis and urticaria have appeared during recent decades. In one of the reports, specific IgE antibodies in sera of four patients who developed asthmatic symptoms after exposure to chloramine-T were demonstrated using a radioimmuno-assay. Three cases of bronchial asthma in workers who had handled chloramine-T powder are described in the present report. Positive skin-prick test reactions to chloramine-T were observed and specific IgE antibodies to human serum albumin treated with chloramine-T were detected using the classic radioallergosorbent (RAST) technique in all three patients.
Adult↗
Pathological and morphometric assessment of testicular parameters in patients with metastatic prostate cancer following treatment with either the antiandrogen Casodex (ZM176,334) or bilateral orchidectomy.
Casodex is an orally active non-steroidal antiandrogen that is highly selective for androgen receptors in animals and man. It is indicated for the non-surgical treatment of advanced prostate cancer in man. The present open controlled study in 13 Casodex-treated and 21 orchidectomy-alone (control) patients addressed the hypothesis that chronic administration of antiandrogens will result in Leydig cell hyperplasia as a result of feedback inhibition of the pituitary resulting in increased luteinising hormone (LH) stimulation of Leydig cells. Although Casodex has been shown to produce a moderate rise in circulating plasma testosterone concentration on chronic treatment in prostate cancer patients, a controlled histopathological and morphometric assessment of the testis following orchidectomy in relapsed Casodex patients showed no effect on Leydig cell populations compared with an orchidectomy alone (control) group. No evidence for induction of Leydig cell hypertrophy or hyperplasia as a result of chronic oral administration of 50 mg Casodex daily was obtained in this study.
Androgen Antagonists↗
Kinetics of sodium current and gating current in the frog node of Ranvier.
The experiments were done on voltage-clamped nodes of Ranvier of the frog. The aim was to study the kinetics of sodium current INa and gating current Igat over a large potential range (-92 to -12 mV) and to compare the time constants for the turning-on of INa or Igat with those for the turning-off measured at the same potential. Sodium tail currents were recorded at different postpulse potentials. Inactivation was inhibited by a few min treatment with 0.5 mM chloramine-T (Wang 1984). The sodium permeability was activated by a 0.4 ms pulse from holding potential (-92 mV) to about 0 mV. At the peak of INa the membrane was repolarized to postpulse potentials between -92 and -12 mV. At E greater than -60 mV the tail currents decayed with two time constants, tau 1 and tau 2, reflecting presumably the turning-off and the inactivation of the sodium permeability. The relation between tau 1 and postpulse potential was bellshaped with a maximum at -32 mV. The tail currents could also be fitted by the Hodgkin-Huxley equation with the sodium activation variable m raised to the second or third power. At E less than -50 mV tau m off was equal to 2 tau 1 or 3 tau 1, respectively, whereas at E greater than -25 mV tau m off was equal to tau 1. In addition, the time constant of the turning-on of sodium activation m (tau m on) was determined, assuming INa approximately m2 (with a small initial delay) or INa approximately m3 (without an initial delay). At -22 mV and -12 mV the ratio tau m off/tau m on was close to 1.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Behaviour of chemically modified sodium channels in frog nerve supports a three-state model of inactivation.
Voltage clamp experiments were done on single myelinated nerve fibres of the frog, Rana esculenta, with 10 mM TEA+ in the external solutions to block potassium channels. Sodium current inactivation was measured in TEA-Ringer solution and after treatment with Anemonia sulcata toxin II (5 microM), internal iodate (20/40 mM), glutaraldehyde (10 mM), chloramine-T (0.6 mM), and 2,4,6-trinitrophenol (1 mM). The diphasic inactivation time course, observed in untreated membranes, is slowed by all these agents in a very similar way. Both time constants are increased and the proportion of inactivation components is changed favouring the slowly inactivating one. Trinitrophenol only slows inactivation, whereas in Anemonia toxin II, internal iodate, glutaraldehyde and chloramine-T inactivation becomes incomplete, so that a persistent current is flowing during depolarizations. None of these agents even at high concentrations however, totally removes inactivation. These modifications of inactivation time course are interpreted as changes of rate constants in a three-state inactivation model with one open and two closed states (o-c-c). After chemical treatment the access to the closed states is impeded and the transitions into the open state are accelerated. If the membrane is depolarized during drug application chloramine-T fails to modify inactivation. The curve relating the steady state inactivation parameter, h infinity, to the conditioning potential, V pp becomes non-monotonic in chloramine-T, i.e. dh infinity/dV pp greater than 0 for V pp greater than 60 mV. Trinitrophenol, which per se fails to produce a persistent current component, increases the persistent current in a fibre pretreated with chloramine-T.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Chloramine-T-induced modifications of K+ channel inactivation in neuroblastoma cells.
Explore the source record for details and available documents.
Animals↗
Effects of chloramine-T on charge movement and fraction of open channels in frog nodes of Ranvier.
The effects of chloramine-T, a substance which partially and irreversibly inhibits inactivation of the Na current (Wang 1984), on the on and off charge movement (Qon and Qoff) and on the relative position of the Qon-E and F-E curves were studied on voltage clamped frog nodes of Ranvier. The decrease of Qoff with increasing pulse duration which is seen in normal fibres was much less pronounced in chloramine-T-treated fibres, i.e. chloramine-T inhibited charge immobilization. Also, the decrease of the time constant tau off which normally accompanies the decrease of Qoff was absent in chloramine-T-treated fibres. A correlation (r2 = 0.80) between the relative tau off and the relative Qoff was observed in chloramine-T-treated fibres. The voltage dependence of the on charge movement, Qon-E, was compared with the voltage dependence of the fraction of open channels, F-E. In normal fibres, the F-E curve rose very steeply and crossed the normalized Qon-E curve so that F greater than Qon/Qon max for E greater than -40 mV. After chloramine-T treatment, the F-E curve rose less steeply and remained below the normalized Qon-E curve at all potentials. The effect of halazone was similar to that of chloramine-T but weaker. The observations about the Qon-E and F-E curve are compared with the findings on the squid giant axon.
Action Potentials↗
Action of benzocaine on sodium channels of frog nodes of Ranvier treated with chloramine-T.
Single myelinated nerve fibres of the frog Rana esculenta were voltage clamped in solutions containing 10 mM TEA to block potassium channels. Reduction of peak INa was measured after equilibrating the membrane in solution containing benzocaine between 0.025 and 2.0 mM. A Hill plot of this effect revealed a slope nH approximately 1 between 0.25 and 1.0 mM but showed deviations for the lowest concentrations (nH less than 1) and the highest concentration (nH greater than 1). Treatment with 0.6 mM chloramine-T irreversibly partially inhibited inactivation of sodium channels leading to a large INa component persisting during depolarization. After treatment the benzocaine effect on peak INa (tested with 0.25-1.0 mM) was unchanged but the persistent component was much more depressed. Benzocaine shifted the steady-state inactivation curve to more negative potentials. This was also observed after chloramine-T treatment which itself produced a curve of decreased slope, shifted to more positive potentials. Recovery from inactivation was studied at different levels of hyperpolarization; it was diphasic in anaesthetic-free solutions before and after chloramine-T treatment although slowed in the latter case. In benzocaine recovery started with a delay (less than 0.6 ms at 16-18 degrees C) and proceeded with a single time constant that decreased with increasing hyperpolarization, was independent of benzocaine concentration and not affected by chloramine-T. The results are compatible with the idea that the affinity of the binding site for benzocaine increases when the channel state changes from resting to open to inactivated with equilibrium dissociation constants of the reaction with resting channels, KR = 0.7 mM and with inactivated channels, KI = 0.04 mM.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Distinctly different rates of benzocaine action on sodium channels of Ranvier nodes kept open by chloramine-T and veratridine.
Single myelinated nerve fibres of the frog, Rana esculenta, were voltage clamped in a fast-exchange chamber in the presence of 10 mM TEA to block potassium channels. After treatment with 0.6 mM chloramine-T for 1-4 min a sizeable INa component persisted even during a 14-s depolarizing impulse. Changing the perfusate to Ringer solution + 1 mM benzocaine resulted in a fast reduction (half time ca. 0.06 s) of the persistent INa, comparable to the rate of block of peak INa during a series of short impulses before chloramine-T. In the presence of 60 microM veratridine the peak INa was followed by a slow exponential (tau s) reincrease of inward current, Is, that did not appreciably inactivate. Application of 0.25 mM benzocaine during a 14-s depolarizing impulse caused Is to decrease exponentially with a large time constant, tau on of 4.3 s. Recovery on washout proceeded with tau off = 3.4s. Tau on was little dependent on benzocaine concentration and was 4.5 s on the average in 1 mM. Tau on in 25 microM was insignificantly (15%) larger than in 1 mM if tested on the same fibre. After equilibration in 25 microM, 0.25 mM and 1 microM, Is(t = 14s) was reduced to 0.69, 0.30, and 0.10, respectively, of the value without anaesthetic. Cooling by only 4-5 degrees C reduced Is and much increased tau s. tau on (1 mM benzocaine) increased almost in proportion to tau s. Tail currents during a series of pulses (1.1 s every 2.5 s) were reduced by 0.25 mM benzocaine clearly faster (tau on = 1.3 s) than Is during a long pulse of the same amplitude.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
The mechanisms of sodium current inhibition by benzocaine in the squid giant axon.
(1) The effects of benzocaine on the ionic currents in the voltage-clamped squid giant axon have been examined under various conditions; intact axons, axons internally perfused with CsF and axons dialysed with tetraethylammonium ions were used. (2) Both the steady state outward (potassium) current and the early transient (sodium) current were reduced by ca. 50% by benzocaine (1 mM). (3) Plots of the changes produced by benzocaine (1 mM) in the Hodgkin-Huxley parameters for the steady state activation (m infinity), the steady state inactivation (h infinity) and the time constants (tau m and tau h) for activation and inactivation of the sodium current are shown. The m infinity and h infinity curves are shifted in positive and negative directions respectively on the voltage axis. The time constants are not greatly affected. (4) In axons in which the sodium current inactivation had been largely removed by treatment with chloramine T, the sodium current was still reduced by ca. 50% by 1 mM benzocaine and the positive shift in activation remained unchanged. (5) The dependence on benzocaine concentration (for less than or equal to 2 mM) of the peak sodium current reduction and the shift in steady state inactivation have been determined. (6) It is concluded that in the squid axon the effects on inactivation are not the main reason for the reduction of the sodium current by benzocaine and that, in common with many other neutral anaesthetics, there are at least two sites at which benzocaine acts.
Animals↗
The existence of a highly tetrodotoxin sensitive Na channel in freshly dispersed smooth muscle cells of the rabbit main pulmonary artery.
To characterize the inward current recorded from single smooth muscle cells of the rabbit main pulmonary artery, a voltage clamp procedure using patch pipettes filled with high Cs solution to inhibit K currents was employed. Under superfusion with normal physiological salt solution, application of a command potential to -10 mV from the holding potential of -80 mV elicited an inward current comprising fast and slow components. In Ca-free solution containing 2.5 mM Mn and 134 mM Na, the major part of the slow inwart current (Islow) ceased, but a transient fast inward current (Ifast) remained. A reduction in the Na concentration in the bath solution inhibited the amplitude of Ifast. Both nicardipine (30 nM) and diltiazem (1-10 microM) inhibited Islow but had no effect on Ifast. Application of tetrodotoxin (greater than 1 nM) in Ca free solution inhibited the amplitude of Ifast in a dose-dependent manner with a dissociation constant of 8.7 nM. Chloramine-T (0.3 mM) increased the peak amplitude and reduced the rate of decay of Ifast and completely inhibited Islow. These results suggest that the inward curent generated in the smooth muscle cells of the rabbit main pulmonary artery is associated with activation of a voltage-dependent Ca channel and a tetrodotoxin-sensitive Na channel.
Animals↗
Removal of rapid sensory adaptation from an insect mechanoreceptor neuron by oxidizing agents which affect sodium channel inactivation.
1. The femoral tactile spine of the cockroach is a mechanoreceptor with a single sensory neuron. The response to a step movement is a burst of action potentials which decays to zero in about 1 s. This rapid adaptation is a property of the action potential initiating region of the neuron. 2. The oxidizing agents chloramine-T and N-chlorosuccinimide selectively and irreversibly remove sodium channel inactivation from neurons in several preparations and are believed to act by oxidation of methionine or cysteine residues in the proteins of the sodium channel. 3. Chloramine-T and N-chlorosuccinimide, applied for a controlled time period, eliminated the rapid adaptation of the tactile spine neuron to an electrical depolarization. After treatment it fired tonically in response to a steady current stimulus. Longer applications of the agents eventually raised the threshold for action potential initiation. 4. Threshold behavior in the tactile spine neuron was characterized by measuring strength-duration relationships for stimulation with extracellular current pulses at the action potential initiating region. The two oxidizing agents caused a voltage-dependent modification of the dynamic threshold properties which led to the change from rapidly adapting to tonic behavior. 5. Two stronger oxidizing agents, N-bromoacetamide and N-bromosuccinimide, raised the threshold of the neuron without removing rapid adaptation. These two agents act similarly to chloramine-T and N-chlorosuccinimide on sodium inactivation in other neurons but are believed to oxidize the tryptophan, tyrosine and histidine residues of proteins in addition to cysteine and methionine.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Iodination of myofibrils and myosin.
The relative reactivity of the tyrosine side chains in the proteins of skeletal muscle myofibrils was determined using iodination techniques. The destruction of ATPase activity of myofibrils and myosin by lactoperoxidase and chloramine-T iodination could be prevented by the attachment of cysteamine to the sulphydryl groups prior to the iodination reaction and subsequent regeneration with thioglycolate or dithiothreitol. Iodination using 1,3,4,6-tetrachloro-3 alpha, 6 alpha-diphenylglycoluril did not require cysteamine treatment for retention of full enzymatic activity. The specific activity of the different proteins varied markedly with desmin, troponin-T, and tropomyosin having the highest labelling with all three iodination procedures. In contrast the myosin light chains had low specific activity when labelled in myofibrils or intact myosin. The isolated light chains, however, were much more highly iodinated. It appears that iodination may be a useful technique for examining protein-protein interactions in the myofibril.
Adenosine Triphosphatases↗
Presence of serum modulates expression of complement receptor type 1 (CR1) on human granulocytes after quartz exposure.
We have investigated the interaction between granulocytes and quartz with respect to the expression of complement receptor type 1 (CR1) and the presence of normal human serum (NHS). Quartz down-regulates selectively CR1 on activated granulocytes. This down-regulation is abolished in the presence of both NHS and heat-inactivated NHS (NHS56) but not human albumin. When quartz was preincubated with NHS (quartz-NHS) before exposure to activated granulocytes, a down-regulating effect was observed in contrast to preincubation with NHS56, which did not induce a down-regulation. Preincubation with cytochalasin B reduced the down-regulation of quartz-NHS, indicating a cytoskeleton-dependent internalization of the receptor. The serine protease inhibitor PMSF partly reduced this down-regulation. Our results indicate that the presence of NHS in the alveolar space influences the interaction between quartz and recruited granulocytes with respect to CR1 expression. Since CR1 is an important opsonin receptor and soluble CR1 can modulate the inflammatory response, this may be of importance in the inflammation and fibrosing process induced by quartz in the alveolar space and lung interstitium.
Blood Physiological Phenomena↗
Studies on the iodination of a ras protein and the detection of ras polymers.
Several methods for the iodination of recombinant v-H-ras protein were compared. The Iodobead method gave greatest incorporation of radioactivity with minimal modification of the ras protein. Upon treatment of the ras protein with [125I] Nal and an Iodobead, radioactivity was initially incorporated into a 22 kDa species with a pl of 5.2, then predominantly into a 23 kDa species with a pl of 5.4. The specific activity of [125I]ras was 6 x 10(6) cpm/pmol total ras protein. Iondination did not alter the biological activity of the ras protein as judged by its ability to bind GTP gamma S and induce maturation of Xenopus laevis oocytes. It is concluded that while iodination alters the apparent molecular weight and pI of ras, presumably by the oxidation of one or more classes of amino acids, this does not affect the biological function of the protein. The ras protein, radioactively-labelled with iodine using the Iodobead method, should be suitable for studies of protein-protein interactions involving ras. Treatment of iodinated ras with the chemical cross-linking agent disuccinimidyl suberate revealed the presence of several minor high molecular weight protein species. This result shows that, in a dilute solution of purified ras protein, the monomeric form is in equilibrium with small amounts of polymeric forms.
Animals↗
Expression of elastinolytic activity among isolates in Aspergillus section flavi.
A survey of the distribution of elastinolytic potential among 32 culture collection isolates of Aspergillus flavus. A. oryzae, A. parasiticus, A. sojae, A. nomius, and A. tamarii revealed this character to be highly conserved within Aspergillus Section Flavi. Furthermore, 144 isolates of A. flavus from environmental samples from six separate regions of the United States produced elastase on solid medium. Most previously described polymorphisms in elastinolytic potential were attributed to the toxicity of borate buffers. Replacement of borate with HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) resulted in detection of elastase production on solid medium by all tested fungal isolates except two that had been in culture over 50 years. In liquid culture, only isolates of A. flavus, A. tamarii, and A. oryzae accumulated elastase activity. Although isoelectric focusing revealed only one isoform (pI 9.0) of elastase in these culture filtrates, elastinolytic activity in filtrates was partially inhibited by both 1,10-phenanthrolene (2 mM) and phenylmethylsulfonylfluoride (2 mM), suggesting the presence of both metallo and serine elastinolytic proteinases.
Aspergillus↗