Andris Kaminskis

The effects of blood flow and detoxification on in vivo cholinesterase inhibition by soman in rats

The in vivo time course of cholinesterase inhibition was measured in brain, lung, spleen, hind limb skeletal muscle, diaphragm, intestine, kidney, heart, liver, and plasma of rats receiving 90 micrograms/kg soman, im. This dose of soman produced severe respiratory depression and transient hypertension, but no significant changes in the cardiac output or heart rate of anesthetized rats. The rate and maximal extent of in vivo cholinesterase inhibition by soman varied widely among the tissues. Although cardiac output was unchanged by soman administration, the blood flow in heart, brain, and lung (bronchial arterial flow and arteriovenous shunts) was increased, whereas blood flow in spleen, kidney, and skeletal muscle was decreased. The relative importance of tissue blood flow, tissue levels of cholinesterase and acetylcholinesterase, and tissue levels of soman-detoxifying enzymes (diisopropyl-fluorophosphatase and carboxylesterase) in determining the in vivo rate and maximal extent of cholinesterase inhibition was examined by multiple regression analysis. The best multiple regression model for the maximal extent of cholinesterase inhibition could explain only 63% of the observed variation. The best multiple regression model for the in vivo rate of cholinesterase inhibition contained three independent variables (blood flow, carboxylesterase, and cholinesterase) and could account for 94% of the observed variation. Of these three variables blood flow was the most important, accounting for 79% of the variation in the in vivo rate of cholinesterase inhibition. This suggests that it may be possible to use a flow-limited physiological pharmacokinetic model to describe the kinetics of in vivo cholinesterase inhibition by soman.

Enhancement of drug absorption after administration by an automatic injector

The effectiveness and rapidity of drug absorption after its administration by an autoinjector was studied in normal male adults. In 12 who received pralidoxime chloride (600 mg) by an autoinjector and by needle and syringe, the plasma concentration of oxime was 82 % higher at 3 min after autoinjector administration, and a therapeutic plasma concentration (4 μg/ml) was attained in about 70% of the time required after needle- and- syringe administration. Atropine given to six subjects by both methods produced tachycardia faster and the time of maximal tachycardia was sooner after autoinjector administration. It is theorized that faster drug absorption occurs because the autoinjector “sprays” the drug through tissue.

Plasma Free Cyanide and Blood Total Cyanide: A Rapid Completely Automated Microdistillation Assay

Techniques are presented which provide direct measurement of both free cyanide (CN-) in plasma and total CN- in whole blood. Loss of total CN- from blood is prevented by conversion to cyanmethemoglobin. Both free and total CN- are assayed by a completely automated method providing readout 17 minutes after sampling. No prior isolation technique is required and sensitivity is adjustable to cover a broad range of CN- concentrations from 1 to 4000 uM. Precision of blood CN- values from 2 to 2500 uM is within +/- 2.3%. No interference results from thiocyanate or thiosulfate at a concentration of approximately 1 mM.

The stability of sarin and soman in dilute aqueous solutions and the catalytic effect of acetate ion

The hydrolysis of aqueous solutions of organophosphonate compounds such as sarin and soman is a critical factor in biochemical and therapeutic studies. The ability to dilute and transport these compounds with assurance of their purity is imperative to the integrity of such studies. An enzymic method has been used to measure low levels of organophosphonates. Maximum stability data were determined in aqueous and isotonic saline solutions. Acetate ion was found to significantly accelerate hydrolysis.1

Carbamoylated Enzyme Reversal as a Means of Predicting Pyridostigmine Protection Against Soman

The inhibition of human and mammalian red blood cell (RBC) Cholinesterase (AChE) in whole blood in the presence of added pyridostigmine has been examined. After the addition of pyridostigmine to animal and human blood, red cells were separated from plasma at varying intervals and their enzyme activity measured. An apparent rate constant (ke) was derived for the reaction sequence in which carbamate is released from AChE inhibited by pyridostigmine. The constant is a complex of the rates of decarbamoylation and reinhibition of AChE in the blood sample. Rate constants were also determined for the spontaneous reactivation (ks) of carbamoylated AChE in the species studied. Values of ks were greater than ke in corresponding species but varied little between species. Pretreatment of animals with pyridostigmine is known to be an effective therapy against organophosphorus compounds, including soman. The ranking of ke values in mammalian blood was the same as that for the protection against soman in animals: monkey>guinea‐pig>rabbit>rat (ke = 0·15, 0·07, 0·05, 0·02 h−1, respectively). Since ke for human blood (0·20 h−1) was greater than that of monkey, pyridostigmine pretreatment would be expected to be an effective prophylaxis for soman in humans.

A Completely Automated Fluorometric Blood Cyanide Method: A Specific Assay Incorporating Dialysis and Distillation

Discrete blood samples can be assayed at a rate of 20/hr. By means of a double lumen catheter, venous or arterial blood can be monitored continuously.

Effects of Physostigmine on the Cardiopulmonary System of Conscious Pigs

Physostigmine, as a pretreatment candidate for nerve agent poisoning, was examined for cardiopulmonary side effects. Cardiovascular and pulmonary parameters were monitored in unanesthetized domestic pigs which received pulmonary arterial infusion of 5 micrograms/kg/min physostigmine salicylate for 2 hr. A level of 74% inhibition of red blood cell (RBC) acetylcholinesterase (AChE) activity was attained in 45 min, and this level of carbamylation increased only slightly during the remaining infusion period. In addition to this large change in AChE activity, minor changes were observed in hematocrit, heart rate, body temperature, mean aortic pressure, pulmonary arterial wedge pressure, and pulmonary artery pressure. Typically, these parameters showed a trend toward elevated levels. Blood gases, pH, respiratory rate, tidal and minute volume, cardiac output, nonelastic resistance, and dynamic compliance were not significantly different from baseline values. The unanesthetized pig responds to physostigmine in a manner similar to that reported for other species and appears to be a suitable model for evaluating cardiopulmonary effects of cholinesterase inhibitors.