Feasibility of cellular adoptive immunotherapy for Epstein-Barr virus-associated lymphomas using haploidentical donors.

Epstein-Barr virus (EBV)-associated lymphomas following bone marrow or solid organ transplantation are often sensitive to immunomodulatory therapies. These have included withdrawal or reduction in immunosuppressive therapy in the solid organ transplant setting and adoptive cellular therapies in the bone marrow transplant (BMT) setting. We describe a strategy for generating EBV-specific cytotoxic T cell therapy lines with substantial killing activity against haploidentical targets. Weekly stimulation of peripheral blood mononuclear cells (PBMCs) for 3 weeks with the irradiated cells of an autologous EBV-transformed B lymphoblastoid cell line (B-LCL), followed by stimulation in the presence of IL-2, yielded T cell lines that were cytolytic for haploidentical B-LCLs but did not lyse haploidentical targets not expressing EBV antigens.

Common shoulder injuries among athletes: evaluation and management.

Shoulder injuries in athletics, especially throwing sports, are a relatively common phenomenon. Assessment and evaluation of shoulder problems require a systematic approach that should be both comprehensive and efficient. This article outlines a sequential process of evaluation that incorporates the following different components of a thorough examination: taking a history, inspecting the shoulder, palpating the structures, assessing active and passive range of motion, testing strength, and performing special tests. Guidelines for management of the most common diagnoses are included.

The effects of antishock trouser inflation during hypothermic cardiovascular depression in the canine model.

The purpose of this study was to investigate the value and to identify any deleterious effects of antishock trouser use during hypothermic cardiovascular depression. Thirteen mongrels were made hypotensive by cooling to a core temperature of approximately 27 degrees C. Eight dogs had antishock trousers inflated for one hour and five dogs served as controls. Metabolic and hemodynamic variables were measured at regular intervals during cooling, during trouser inflation, and after trouser deflation. No study animal experienced ventricular fibrillation. Neither central temperature, pH, or serum potassium nor mean arterial BP or systemic vascular resistance were significantly affected by trouser inflation or deflation. Antishock trouser use during the early phase of hypothermia before rewarming does not appear to result in a central bolus of cold, acidotic, hyperkalemic blood or the precipitation of ventricular fibrillation. There appears to be no significant hemodynamic benefit of antishock trouser use early in the management of hypotension caused by moderate hypothermia.

Actions of ethanol on voltage-sensitive sodium channels: effects on neurotoxin binding.

Exposure of rat forebrain synaptosomes (P2) to ethanol in vitro reduced the specific binding of [3H]batrachotoxinin A 20-alpha-benzoate ([3H]BTX-B) to voltage-sensitive sodium channels. This effect of ethanol was concentration-dependent and was affected by the membrane potential. Under depolarizing conditions ethanol was significantly more potent at inhibiting [3H]BTX-B binding. Scatchard analysis of [3H]BTX-B binding revealed that ethanol increased the equilibrium binding constant without affecting the apparent maximum number of binding sites. The rate of formation of the [3H]BTX-B/receptor complex was unchanged in the presence of ethanol whereas the rate of dissociation was accelerated by ethanol. These findings are consistent with an indirect allosteric mechanism for inhibition of [3H]BTX-B binding. The binding of [3H]saxitoxin was unaffected by ethanol suggesting that the specific receptor sites in the channel display differential sensitivity to the inhibitory effect of ethanol. These data, in conjunction with ion flux measurements, provide further evidence that ethanol can affect the voltage-sensitive sodium channels in neuronal membranes.

Actions of ethanol on voltage-sensitive sodium channels: effects of acute and chronic ethanol treatment.

The effects of acute and chronic ethanol treatment on neurotoxin-stimulated 22Na+ uptake and [3H]batrachotoxinin A20-alpha-benzoate binding to neuronal sodium channels were studied in rat forebrain synaptosomes. Fluorescence measurements were used to assess the intrinsic order or fluidity and the sensitivity to ethanol of rat forebrain synaptic plasma membranes at various intervals during and after chronic ethanol treatment. Acute ethanol administration had no significant effect on neurotoxin binding in the absence or presence of ethanol in vitro or on sodium uptake in the absence of ethanol in vitro. However, a single dose of ethanol produced a dose and time-dependent attenuation of the inhibitory effect of ethanol on sodium uptake, suggestive of acute tolerance. Chronic ethanol treatment reduced the influx of 22Na+ in the presence of batrachotoxin and diminished the inhibitory effect of ethanol in vitro on sodium uptake for up to 20 days after withdrawal, but the specific binding of the neurotoxin in the presence or absence of ethanol was unchanged. Synaptic plasma membranes from chronic ethanol-treated rats showed no change in intrinsic order but the disordering effect of ethanol was significantly smaller for up to 20 days after withdrawal. Results of this study demonstrate that brain tissue from ethanol-treated rats can adapt rapidly to some effects of ethanol and that chronic ethanol administration can reduce the effects of ethanol on physical and functional properties of neurons for a prolonged period of time.

Intravenous fluid flow beneath inflated antishock trousers in a canine hemorrhagic shock model.

A study was performed to determine the percentage of fluid that enters the central circulation following infusion in a peripheral vein beneath an inflated pneumatic antishock garment (PASG) during hemorrhagic shock in a canine model. Seven mongrel dogs were bled acutely to a systolic blood pressure of 50 to 60 mm Hg. The PASG then was inflated. One to two microcuries of Technetium-99-labeled macroaggregated albumin was infused into a peripheral vein in the hind leg, followed by a 10-mL/kg fluid bolus. The albumin particles that reached the central circulation were trapped in the pulmonary capillary bed. Following infusion, a full body scan was performed. The percentage of injected fluid entering the central circulation was calculated by comparing the total body counts to counts localized to the thorax. Average mean arterial pressure was 111 mm Hg prior to bleeding, 47 mm Hg prior to shock trouser inflation, and 66 mm Hg after inflation. The percentage of injected Technetium-99-labeled albumin that reached the central circulation ranged from 80.5% to 99.6%, with a mean of 91.3%. We conclude that a significant percentage of IV fluids infused beneath inflated PASG will reach the central circulation.

Effects of ethanol on the functional properties of sodium channels in brain synaptosomes.

Voltage-sensitive sodium channels in excitable cell membranes are responsible for the rapid increase in permeability to sodium ions that occurs during depolarization. Neurotoxins that bind with high affinity and specificity to voltage-sensitive sodium channels have been widely used to identify and characterize the structure and function of sodium channels in nerve and skeletal muscle. This chapter describes the actions of ethanol on the functional properties of voltage-sensitive sodium channels in mammalian brain nerve endings. The effects of acute and chronic ethanol administration are also reviewed. Alterations in the function of neuronal membrane sodium channels may be involved in the depressant effect of ethanol.

Ionizing radiation alters the properties of sodium channels in rat brain synaptosomes.

The effect of ionizing radiation on neuronal membrane function was assessed by measurement of neurotoxin-stimulated 22Na+ uptake by rat brain synaptosomes. High-energy electrons and gamma photons were equally effective in reducing the maximal uptake of 22Na+ with no significant change in the affinity of veratridine for its binding site in the channel. Ionizing radiation reduced the veratridine-stimulated uptake at the earliest times measured (3 and 5 s), when the rate of uptake was greatest. Batrachotoxin-stimulated 22Na+ uptake was less sensitive to inhibition by radiation. The binding of [3H]saxitoxin to its receptor in the sodium channel was unaffected by exposure to ionizing radiation. The effect of ionizing radiation on the lipid order of rat brain synaptic plasma membranes was measured by the fluorescence polarization of the molecular probes 1,6-diphenyl-1,3,5-hexatriene and 1-[4-(trimethylammonium)phenyl]-6-phenyl-1,3,5-hexatriene. A dose of radiation that reduced the veratridine-stimulated uptake of 22Na+ had no effect on the fluorescence polarization of either probe. These results demonstrate an inhibitory effect of ionizing radiation on the voltage-sensitive sodium channels in rat brain synaptosomes. This effect of radiation is not dependent on changes in the order of membrane lipids.

Actions of ethanol on voltage-sensitive sodium channels: effects on neurotoxin-stimulated sodium uptake in synaptosomes.

Exposure of rat brain synaptosomes to ethanol in vitro reduced the neurotoxin-stimulated uptake of 22Na+. This effect of ethanol was concentration-dependent, occurred with concentrations of ethanol achieved in vivo and was fully reversible. The inhibitory effect of ethanol on neurotoxin-stimulated sodium uptake was due to a decrease in the maximal effect of the neurotoxins. Ethanol reduced the rate of batrachotoxin-stimulated sodium uptake when measured at 3, 5 and 7 but not 10 or 20 sec after the addition of 22Na+. In a series of aliphatic alcohols, there was a good correlation between potency for inhibition of batrachotoxin-stimulated 22Na+ uptake and the membrane/buffer partition coefficient, suggesting that a hydrophobic site in the membrane was involved in the action of the alcohols. Ethanol did not affect the scorpion venom-induced enhancement of batrachotoxin-stimulated sodium uptake. The inhibitory potency of tetrodotoxin was also unaffected by ethanol. These results demonstrate that ethanol has an inhibitory effect on neurotoxin-stimulated sodium influx occurring in voltage-sensitive sodium channels of brain tissue.

Effects of ethanol exposure on brain sodium channels.

Ethanol, like other general anesthetics, probably acts through an interaction with excitable membranes. When ethanol is present in membranes, the lipid structure of membranes is disordered. This effect can lead to alterations in functional entities that require a particular lipid environment. Sodium channels, requiring such an environment, have been studied for their sensitivity to ethanol. In synaptosomes, ethanol in vitro reversibly inhibits sodium influx stimulated by batrachotoxin or veratridine in concentrations equivalent to those found in animals during ethanol intoxication. Only the maximum stimulation of the toxins is altered by ethanol. The potencies of aliphatic alcohols are directly related to their lipid solubilities. Acute and chronic ethanol administration reduces the effectiveness of ethanol in vitro. This tolerance dissipates after a single dose of ethanol as it is eliminated from the body. However, after 4 days of ethanol treatment, the tolerance lasts for over 20 days. These data suggest that ethanol might disrupt one of the basic processes of neural function by acting directly on the environment of the sodium channel. The channel appears to be able to adapt to the presence of ethanol through the induction of tolerance.

Ethanol inhibits veratridine-stimulated sodium uptake in synaptosomes.

The effects of ethanol and pentobarbital on voltage-sensitive sodium channels in whole brain (rat) synaptosomes were studied using isotopic flux measurements. Incubation of synaptosomes with ethanol or pentobarbital in vitro inhibited veratridine-stimulated 22Na+ uptake. The effect of ethanol is dose-dependent, occurs at sublethal, pharmacologically relevant concentrations and is fully reversible. These results suggest that ethanol and pentobarbital directly interfere with sodium channel function in nervous tissue. Alterations in sodium channel function may be a possible mechanism for the central nervous system (CNS) depressant action of ethanol and related compounds.

Alterations in dopaminergic function after subacute ethanol administration.

The in vivo and in vitro effects of d-amphetamine were assessed in rats after subacute (24 hr) administration of ethanol vapor (28 mg/l). Forty-eight hours after the termination of ethanol vapor inhalation, when the animals were tolerant to the hypothermic effect of ethanol, a decreased hypothermic response to d-amphetamine was detected, suggesting that functional changes in dopaminergic activity might be associated with ethanol tolerance. The hyperthermic effect of d-amphetamine was similar in the ethanol-tolerant and control animals. The d-amphetamine-stimulated release of [3H]dopamine from crude synaptosomal (P2) fractions of striatum was nearly abolished immediately after subacute ethanol administration. Samples prepared from ethanol-tolerant animals released less [3H]dopamine in response to d-amphetamine perfusion compared to samples prepared from control animals when tested 48 hr after ethanol vapor exposure. Subacute ethanol administration appears to produce long-acting changes in d-amphetamine-sensitive dopaminergic mechanisms.

Ethanol and functional tolerance: interactions with pimozide and clonidine.

Inhalation of ethanol vapor (28 mg/l) for 24 hr caused a reduction of body temperature in rats. The mean peak blood ethanol concentration was 3.36 +/- 0.14 mg/ml. When ethanol (2 g/kg i.p.) was administered 48 hr after termination of ethanol inhalation, the ethanol vapor-treated animals maintained normal body temperatures despite the presence of blood ethanol concentrations (1.81 +/- 0.04 mg/ml) which caused hypothermia in control animals. The rate of clearance of ethanol from blood was found to be 0.34 +/- 0.01 mg/ml/hr in ethanol vapor-treated and control animals when ethanol was administered acutely 48 hr after the inhalation period. Animals tolerant to the hypothermic effect of ethanol demonstrated diminished sensitivity to the hypothermic effect of clonidine (50 micrograms/kg s.c.). Pretreatment of naive animals with the dopaminergic blocker, pimozide, significantly attenuated the hypothermic response to acute ethanol administration (2 g/kg i.p.) and potentiated the hypothermic response to acute clonidine administration. A dopaminergic mechanism may partially mediate the reduction in body temperature associated with acute ethanol administration.