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Co-carboxylase in the liver and its formation from vitamin B1 in experimental diabetes. Modification of the manometric method of Ochoa and Peters for estimating co-carboxylase in tissues



Co-carboxylase in the liver and its formation from vitamin B1 in experimental diabetes. Modification of the manometric method of Ochoa and Peters for estimating co-carboxylase in tissues



Acta vitaminol, 5: 3-7



The method of Ochoa and Peters (Abst. 3097, Vol. 8) was modified as follows: (1) the tissue was extracted with 2 volumes of 0.001 N HCl instead of with water; (2) 1 g. of yeast, dried at room temperature, was homogenized with 2 lots of 50 ml. of 0.1 M Na2HPO4 then washed twice with 50 ml. of water, and centrifuged each time, each stage occupying not more than 3 min. ; (3) 50 micro g. instead of 10 micro g. of vitamin B1 was added to each Warburg flask; (4) a blank was run without addition of pyruvate. Normal and alloxan-diabetic rats were injected with 20 mg. vitamin B1 per kg. bodyweight and killed 30 min. later. The average content of co-cartaoxylase, in micro g. per g. fresh tissue, was for normal liver without vitamin B1 9.8 and 9.4, for normal liver with vitamin B1 20.1, for diabetic liver without vitamin B1 4.8 and 4.4, and for diabetic liver with vitamin B1 6.7. The diabetic animals' R.Q. had been determined 6 hr. previously, before, and 60 min. after, injection per kg. bodyweight of 20 mg. vitamin B1 and 5 g. glucose, but only slight changes in it were found.

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