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Biomedical subjects

S Muchnik

Publications and source records attributed to S Muchnik.

At least 37 records · Page 2Linked to original sources

Blockers of potassium current and resting membrane potential in rat muscle fibers.

Rat diaphragm fibers were equilibrated for several hours in 150 mM KCl; when they were returned to 5 mM KCl the resting potential went back to its original level with a half time of 17 min. This repolarization was blocked by 5 mM BaCl2, a blocker of the inward rectifier K channel. On the other hand, 0.1 mM apamin and 0.02 mM glibenclamide which block the Ca-dependent and ATP sensitive K channels, respectively, and 0.1 mM 9-AC a blocker of the Cl- channel did not affect the repolarization. 5 mM barium decreased the K conductance measured under current-clamp conditions in diaphragm muscle fibers. The possible role of the inward rectifier system in the repolarization following return to normal [K]o is discussed.

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The effect of BCG on thoracic duct lymphocytes.

Adoptive immunotherapy is becoming increasingly more important in the management of advanced malignancies. This report describes the results of immunomodulation therapy with BCG in the Dunning tumor. In addition it describes new techniques in the harvesting of lymphocytes. Thoracic duct lymphocytes from 34 rats were evaluated for the effect of the presence of the Dunning R-3327 AT-3 tumor as well as for the response to bacillus Calmette Guerin (BCG). Both tumor and BCG resulted in significant changes in the helper/suppressor T cell ratios.

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Tumor infiltrating lymphocytes: the effect of bacillus Calmette Guerin on helper/suppressor-T cell ratios of treated and untreated tumors.

Copenhagen X Fischer rats bearing single and bilateral Dunning R3327 AT-3 tumors were either treated or not treated at a single site with bacillus Calmette Guerin (BCG). One week later tumors were removed, tumor infiltrating lymphocytes (TIL's) isolated, and then characterized for total-T, helper-T and suppressor-T cell subsets utilizing monoclonal antibodies. The purpose was to determine the effect of BCG on TIL's in treated as well as untreated tumors. In summary: 1) BCG treatment significantly alters TIL distributions at both injected and noninjected sites; 2) a noninjected contralateral tumor compromises the effectiveness of BCG therapy at the suppressor T cell level; 3) contralateral tumors, whether inoculated or not, have similar TIL distributions.

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Efficacy of immunopriming prior to isolation of tumor infiltrating lymphocytes for use in adoptive immunotherapy.

Adoptive immunotherapy is dependent upon the leukocytic subsets isolated as tumor infiltrating lymphocytes (TIL) prior to in vitro expansion with interleukin-2. To favorably influence T-cell subset representation in TIL the efficacy of bacillus Calmette Guerin (BCG) and cyclophosphamide (CTX) priming was evaluated in rats bearing Dunning R3327-AT prostatic tumors. When assessed by immunohistochemistry, both agents significantly (p less than 0.001) increased helper-T representation and decreased that by suppressor-T cells. As a result helper/suppressor (H/S) T cell ratios of TIL from untreated tumors (0.73 +/- 0.11) were significantly (p less than 0.001) elevated by both BCG (1.93 +/- 0.39) and CTX (1.40 +/- 0.25). Immunopriming might enhance adoptive immunotherapy by increasing the H/S ratio of TIL prior to their culture.

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Changes in tetrodotoxin-resistant action potentials after passive transfer of myasthenia gravis patient sera.

Muscle electrical activity has been studied in mice after intraperitoneal injection of sera from myasthenia gravis (MG) patients. Myasthenic serum did not modify the electrical properties of innervated muscle fibres. The resting membrane potential and the action potential parameters remained unchanged. However, tetrodotoxin (TTX)-resistant action potentials of denervated muscles were reduced by myasthenic serum, possibly in association with receptor endocytosis induced by the immunoglobulin. However, a direct effect of MG serum on TTX-resistant sodium channels cannot be ruled out.

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A sequential study of the peripheral nervous system involvement in experimental Chagas' disease.

To search for the sequential compromise of the spinal cord, nerves, and skeletal muscle in mice chronically infected with Trypanosoma cruzi, animals were subjected to electromyographic investigation, end-plate recordings, and histological studies at 7, 15, 37, 60, 90, 120, 180, 270, and 360 days postinfection. Electromyographic studies showed signs of motor unit remodeling as early as 15 days postinfection, when diminished duration and amplitude of motor unit potentials pointing to a primary muscle involvement were found. Thereafter, certain features of denervation, reinnervation, and primary muscle involvement were often found to coexist. Low miniature end-plate potentials with normal frequency and acetylcholine quantum content were found in end-plate recordings made at the phrenic-diaphragm in vitro. Double end-plate potentials were observed in most of the tested muscle fibers from day 90 postinfection. All these features suggest post-synaptic damage of the end-plate and the presence of reinnervation after day 90 postinfection. Histological studies disclosed inflammatory infiltrates consisting of lymphocytes and macrophages, with vasculitis as the main lesion in the hamstring muscles; intracellular parasites were seen in 25% of the cases. Neuropathic features, as expressed by type fiber grouping and grouped muscle fiber atrophy, were found. On nerve examination epineural, perineural, and endoneural vasculitis were seen. Digestion chambers and myelin ovoids (axonal degeneration) were observed. In teased fiber preparations, segmental internodal and paranodal demyelination and remyelination were found. The lumbar inflammatory spinal cord failed to show grey or white matter infiltrates. However, spinal roots and dorsal root ganglia were densely affected by inflammatory cells.

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Effect of passive transfer of myasthenic serum on mechanical, electrical and neuromuscular transmission properties of mouse skeletal muscle.

Neuromuscular transmission, muscle electrical activity and muscle mechanical properties have been studied in mice after the intraperitoneal transference of sera from myasthenic patients. Measurements of mepp's amplitude appear to be a suitable method for myasthenia gravis diagnosis since a high percentage of the sera from these patients caused a decrease in mepp's amplitude. The increase of the premenstrum weakness in myasthenic patients did not appear to be associated with greater fall of mepp's amplitude, since serum obtained in the premenstruum and far from it produced similar results. Myasthenic serum did not modify the electrical properties of innervated muscle sarcolemma. The resting membrane potential and the action potential parameters remained unchanged. However, tetrodotoxin resistant-action potentials of denervated muscles was reduced by myasthenic serum, possibly in association with receptor endocytosis process induced by the immunoglobulin. Muscle mechanical properties did not show alterations, even in the presence of positive titer of anti-striated muscle antibody.

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Tools to differentiate immunologic and non-immunologic myasthenia gravis in infancy.

Twelve patients between 15 months and 13 years of age with clinical and pharmacological features of myasthenia gravis were studied. Repetitive nerve stimulation did not offer valuable information; the patients demonstrated either inspecific muscular decremental response or had normal behavior. Two clear groups of patients were identified after measurements of acetylcholine receptor (AChR) antibodies and MEPP amplitude recorded in the diaphragm of mice injected with sera from those patients. The first group included patients with positive AChR antibodies titers and decreased MEPP amplitude. The second one had negative AChR antibodies titers and MEPPs with normal amplitude. These data strongly suggest immunologic and non-immunologic mechanisms for the former and later respectively.

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Effects of resting membrane potential and intactness of the T-tubules on caffeine contractures in rat skeletal muscle.

We have studied the effects of changes in the resting membrane potential (Vm) and T-tubules on caffeine contracture (25 mM) elicited in rat soleus muscle in vitro at 34 degrees C. In high [K]o (30-140 mM, [K]o X [Cl]o constant) caffeine contractures were reduced by about 40-50% and had a faster time course than in normal Krebs ([K]o = 5 mM). Detubulation of the muscles by an osmotic treatment produces a reduction of about 30% in the caffeine contracture tension. Our results with high K solutions suggest a reduced sensitivity of the myofibrils to calcium released by caffeine. The effects of detubulation on caffeine contracture suggest that caffeine may have a direct effect on sarcolemma in addition to its well known action on the sarcoplasmic reticulum (SR). However, a depletion of the calcium content in the SR of depolarized muscle fibres as well as an anatomical damage produced by the osmotic treatment can not be ruled out as an explanation for the reduced caffeine contracture.

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Mechanical and electrical properties of denervated rat skeletal muscles.

Mechanical activity (twitch and tetanus) and electrical activity (single and repetitive action potentials) were recorded in vitro (34 degrees C) in control and denervated (3 to 14 days) soleus and extensor digitorum longus muscles of the rat. After denervation tetanic tension (100 to 200 Hz, 500 ms duration) was decreased in both types of muscles. Denervation reduced significantly the rates of rise and fall and the amplitude of the action potential in both types of muscle fibers. In denervated fibers with very low resting membrane potential no action potentials could be recorded: in these fibers only a slow response without overshoot was detected. Hyperpolarization of denervated fibers to -90 mV prior to application of the depolarizing pulse increased their excitability. Action potential amplitudes were well maintained during tetanic stimulation (200 Hz, 40 to 90 ms) in innervated fibers. Depolarization of the innervated fibers with cathodic current before the tetanic pulse hindered the generation of repetitive action potentials at 200 Hz. A proportion of denervated fibers stimulated at 100 to 200 Hz generated only one action potential or gave rise to an incomplete train. Hyperpolarization of the denervated fibers resulted in an improvement in the ability to generate a train of action potentials at 100 to 200 Hz. A group of denervated fibers exhibited well maintained action potentials during tetanus. We suggest that failure in the repetitive electrical activity of denervated fibers could be the reason for the reduced tension of tetanus. Depolarization of the fibers and/or the increment in the electrical time constant of the sarcolemma are suggested for the decrease in the electrical excitability of denervated fibers.

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Caffeine contractures in denervated slow-twitch and fast-twitch muscles of the rat.

Caffeine contractures (25 mM) and twitches were registered in vitro (34 degrees C) in normal and denervated soleus and extensor digitorum longus of the rat. Both muscles lost weight progressively after denervation although the loss of weight in soleus muscles was more manifest. Denervated extensor digitorum longus muscles showed an increase in the caffeine contracture tension (expressed in g/g muscle) whereas the response generated by denervated soleus muscles was smaller than the control values. The time to peak of the caffeine responses was shortened in both types of muscles after denervation. Twitches were larger and had a slower time course than in normal muscles, especially in the extensor digitorum longus muscles. These findings suggest that some changes in the excitation-contraction coupling could be responsible for the potentiation of the twitches in denervated muscles.

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The effect of verapamil and Ca free solution on mechanical and electrical properties in fast twitch mammalian skeletal muscle.

The effects of verapamil (0.01-0.1 mM) and Ca-free solutions (0 Ca, 3 mM MgCl2, 5 mM EGTA) upon mechanical and electrical properties of a fast twitch mammalian skeletal muscle (extensor digitorum longus) have been studied. Ca-free saline reduces twitch and tetanus tension. The response to single pulses was not affected in the presence of verapamil, but as the rate of stimulation increased, peak tensions were lower and a gradual decrease of tension was observed: frequency-dependent effect. Caffeine (30 mM) contracture was significantly reduced by both solutions. The fibers in Ca-free saline were depolarized (6 mV) and +dV/dt, -dV/dt and overshoot of the AP's were reduced. On the other hand, verapamil did not affect Vm. The presence of 0.1 mM verapamil hindered the ability of the fibers to generate AP's at high rates of stimulation. Lower concentration of verapamil (0.01) did not affect the repetitive electrical activity, but tetanic tension was decreased. These findings support partially the hypothesis about the dependence of mammalian skeletal muscle from external Ca. Verapamil would have several mechanisms of action: a) Ca-channel blocker action, b) local anaesthetic effect, and c) reduction of Ca release from the sarcoplasmic reticulum.

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Effects of actinomycin D on contractile properties of denervated rat skeletal muscle.

We studied the mechanical and electrical properties in the extensor digitorum longus muscle of Wistar rats in vitro 4 days after denervation. In a group of animals, actinomycin D was injected intravenously at the time of denervation. The prolongation of contraction and relaxation time, and the increase in twitch tension observed in denervated muscles were prevented by the drug. Other denervatory changes such as the elimination of cooling potentiation, posttetanic potentiation, staircase phenomenon, and shortening of caffeine contracture were also inhibited or partially reverted by actinomycin D. In addition, some electrical denervatory changes such as the decrease of resting membrane potential and slowing of maximum rate of fall of the action potential were partially abolished by actinomycin D. These results support the hypothesis that a new synthesis of proteins is related to denervation changes.

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Co2+, low Ca2+, and verapamil reduce mechanical activity in rat skeletal muscles.

We have studied the effects of Co2+ (5 mM), low-Ca2+ solution [0 added CaCl2, 5 mM ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid, 3 mM MgCl2 Ringer], and verapamil (0.1 mM) on mechanical and electrical properties of rat soleus muscle in vitro at 34 degrees C. Muscle fibers had normal resting potentials in Co2+ and verapamil solutions. Low-Ca2+ solution produces a depolarization of approximately 4 mV. The action potentials are smaller and have a slower time course when exposed to test solutions. Iterative generation of action potentials in the presence of Co2+ and low-Ca2+ solution is not modified. In the presence of Co2+ or low-Ca2+ solution, the mechanical output, twitch and tetanus tensions, and caffeine contracture are reduced significantly. Verapamil produces a decrease in the twitch and tetanic tensions but does not modify the caffeine contracture tension. The effect of verapamil on the twitch becomes more manifest when the muscle is stimulated at 3-5 Hz. We suggest that changes in the action potential characteristics or the inhibition of a Ca2+ current are responsible for the mechanical changes observed in the presence of the drugs.

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