131 



running times were not as constant as in previous work with 

 Hajdu.2 



The results seemed positive in the case of "Bent Horn," 

 "Shorty," and "Spott}%" three young goats, about 15 months old. 

 The result was negative with "Nanny," an old goat which be- 

 longed to the original flock received in full grown condition four 

 years earlier. Figure 28 suggests the presence of an active substance 

 in our extracts, capable of alleviating or abolishing the symptoms 

 of myotonia. The extracts had no acute effect and developed their 

 action gradually. This action outlasted the injections and disap- 

 peared gradually. The active matter present thus did not act as 

 drugs do, which act the strongest when their concentration is the 

 highest. 



The myotonic goat is not an ideal test object. The size and smell 

 of the animal are bigger than desired; the expense and labor in- 

 volved in keeping it are considerable. The measurements are time 

 consuming. So the final isolation of the active agent, if any, will 



the clear liquid concentrated in vacuo at 20-23 °C in two steps to 1.4 liter. 

 The fluid was stored overnight at — 20°C, on thawing the precipitate re- 

 moved on a small Sharpies centrifuge. Four volumes of methanol were added, 

 the precipitate separated, the fluid stored at — 20°C, and the newly formed 

 precipitate separated again. The fluid, when liberated from its methanol 

 content, was poisonous to mice, probably owing to its potassium content. 

 Concentrated sulphuric acid was added gradually under strong mixing, 

 whereupon a white crystalline precipitate formed, presumably consisting of 

 K2SO4. H2SO4 was added until the addition of 10% sulphuric acid caused no 

 appreciable precipitate. The pH at this state was about 5. 



The solution was concentrated in vacuo to 500 ml. Two liters of acetone 

 were added, the precipitate separated, and shaken out with acetone. The 

 supemates were rejected and the semi-liquid precipitate redissolved in 500 

 ml water. This fraction was injected after neutralization, a small quantity of 

 penicillin being added to prevent infection. 



' On the first three days of the treatment, the running time was measured 

 one hour before and one hour after the injection. The two measurements 

 showed no marked difference so it was concluded that the injections had no 

 acute effect and subsequently the running times were measured only before 

 the injections. 



