Experiments on human beings disclose a con

nexion between glycaemic and alcoholaemie variations which appears to support the view that sugar in the organism ultimately undergoes alcoholic scission.

T. H . Po p e. _ A ction of n arcosis on the ch em ical c o m p o si­

tion of th e brain. M. Se r e j s k i (Biochem. Z., 1927, 1 8 2 , 188—192).—Chloroform narcosis in dogs leads to an increase in the lipin content of both the white and the grey matter of the brain. The white matter shows a greater increase in cholesterol and unsatur­

ated phosphatides than the grey matter. The total nitrogen of the white and grey matter is not sig­

nificantly altered, but there is a slight tendency to

increase. J. Pr y d e.

T oxicity of lo ca l anaesthetics ad m in istered intra-arterially. T. Ku r o d a (Biochem. Z., 1927, 1 8 1 , 172—175).—The toxicity of anaesthetics of the cocaine group is less after intra-arterial than after intravenous administration, probably owing to a detoxication during passage through the capillary plexus. C. R. Ha r in g t o n.

B lood reaction s of the alkaloids of Ceanothus americanus. C. E. T h a r a l d s e n and J. K r a w e t z (Amer. J. Physiol., 1927, 79, 545—552).—Both in vitro and when administered orally or subcutaneously, these alkaloids reduced the clotting time of blood.

Their influence was probably on the action of the thromboplastin. R . K . G a n n a n .

[Pharm acology of] quinine. G. We i s s and R. A. Ha t c h e r (J. Pharm. Exp. Ther., 1927, 30, 327—333).—The toxicity of quinine is dependent on the rate of its intravenous injection. For the cat, the fatal dose of the hydrochloride falls from 140 mg.

per kg. body-weight to 100 mg. as the rate of injec­

tion is increased from 2 to 5 mg. per kg. body-weight per min. Cats recover within about 3 hrs. from the toxic effects of 70% of the fatal dose. The exclusion of the kidneys from the circulation has little influence on the rate of recovery, and perfusion experiments show that the greater part of an intravenous dose is probably destroyed in the liver. E. A. Le n t.

[Pharm acology of] quinine. R. A. Ha t c h e r

and H . Gold (J . Pharm. Exp. Ther., 1927, 3 0 , 347—

350).—Quinine is eliminated practically completely from human blood in 6— 2 0 min. after intravenous

injection. E. A. Lu n t.

[P harm acology of] q u i n i d i n e . S . We i s s and R- A. Ha t c h e r (J. Pharm. Exp. Ther., 1927, 3 0 , 335—345).—The toxicity of quinidine sulphate varies with the rate of intravenous injection. The average fatal dose for the cat, when injected at the rate of 5 mg. per kg. body-weight per min., is about 80 mg.

per kg. About 95% of an intravenous dose leaves

the blood within 5 m in .; the elimination is practically complete in 3—4 hrs. Perfusion experiments indicate that the liver of the cat destroys quinidine and quinine at practically the same rates. The quinidine was determined by titration with bromine water.

E. A. Lu nt. N e p h elo m etric d ete rm in a tio n of s m a ll quan­

titie s of a rsen ic. M . De l a v il l e and J . Be l i n

(Bull. Soc. Cliim. biol., 1927, 9 , 91—93).—A mixture of 5 c.c. of the fluid investigated with 5 c.c. of a reagent containing sodium hyposulphite (1 0%) in hydrochloric acid is boiled and cooled, and the tur­

bidity compared in a nephelometer with that of a standard arsenic solution. The error is ± 6%, using solutions containing 0-0005 mg. of arsenic per c.c.

In biological fluids, organic matter is destroyed first by a mixture of nitric and sulphuric a cid s; the liquid is then evaporated and diluted. L. F. He w i t t.

D etectio n of s m a ll q u an tities of lea d in o rg a n s b y ch em ica l and sp ectro g ra p h ic m e a n s. M.

Kl o s t e r m a n n (Ver. Ges. deut. Naturforsch. Aerzte, 1926, 3 , 1116— 1118).—Lead is obtained as dioxide by ordinary methods, and the formation of a deep blue colouring matter by the interaction of tetra- methjddiaminodiphenyhnethane, lead dioxide, and glacial acetic acid gives a colorimetric criterion for the quantity of lead present. The method has been calibrated by the addition of known amounts of lead to various organs and determining its quantity by the procedure described. The lead may be recon­

verted into the dioxide and then determined spectro- graphically. R . A. Mo r t o n.

K in etics of th e a ction of ca rb o x y la se. E.

Ha g g l u n d and T. Ro s e n q u is t (Biochem. Z., 1927, 1 8 1 , 296—301).—The optimal phosphate concen­

tration for the extraction of carboxylase from yeast is 0-3—0 -3 5if, such extracts being four times as active as an aqueous extract. The variation of reaction velocity with concentration of substrate and with amount of enzyme is studied.

P. W. Cl t jt t e r b u c k. C atalase. I. H. v o n Eu l e r and K . Jo s e p h s o n

(Annalen, 1927, 4 5 2 , 158— 181).—B y a modification of the method of Hennichs (A., 1926, 756), the authors have obtained an exceedingly active prepar­

ation of catalase from horse-liver. The first extraction of the liver -with water does not remove all the enzyme, a second extraction yielding a solution practically identical with the first. Only a single precipitation of the combined extracts with a half volume of alcohol should be employed. The solution (C.F. 1500) so obtained was adsorbed with aluminium hydroxide and the enzyme eluted with 0-02A7- ammonia solution, 46% of the activity being obtained.

Dialysis of this solution (C.F. 7500) for 60 hrs. resulted in no loss of activity, and yielded a product 'with C.F. 14,600. Elution with sodium hydrogen phos­

phate removes oidy 29% of the activity together with a much larger amount of blood pigment, and in this case a loss of activity occurs during dialysis, the final product having C.F. 4600. After dialysis of the solution obtained from the ammonia elution, the enzyme was adsorbed on kaolin in a . medium having 5, and again eluted with 0-02i\T-ammonia.

BIO CHEMISTRY. 377 The solution (C.F. 26,000) after dialysis through a

collodion membrane at 0° for 110 hrs. yielded a preparation having C.F. 43,000, which could be concentrated under diminished pressure without loss of activity. In agreement with Hennichs, it was found that there is no relation between the activity of the preparations and the percentage of iron present.

Two preparations having C.F. 43,000 and 40,000, respectively, contained N 14-72 and 15-06%; S 0-78 and 1-25%; Fe 0-63 and 0-15%, respectively (phos­

phorus absent), the ash content of the two samples being only 1-4 and 0-94%, respectively.

J. W. Ba k e r. C ollagen -d issolvin g en zym e (co lla gen ase).

W. S . Sa d ik o v (Biochem. Z., 1927, 181, 267—283).

—Whereas freshly-prepared trypsin does not, many commercial preparations (especially when old) do attack tendon collagen. When minced pancreas is digested with 2 0% glycerol at the ordinary tem ­ perature for 24 hrs., an extract is obtained which attacks both fibrin and collagen, and the gland residue if further digested for 24 hrs. at 40° with glycerol gives an extract which attacks only collagen.

Attempts were made to purify collagenase by adsorp­

tion methods. The enzyme is not precipitated by safranine, but it is adsorbed by kaolin (distinction from fibrinase). Charcoal adsorbs fibrinase partly, but does not adsorb collagenase. Kaolin and char­

coal adsorbates of pepsin contain an enzyme which in acid solution converts collagen into a glue, but cannot further digest it (glutinase). When an old pancreatic solution was fractionated with kaolin and charcoal, whilst with charcoal no selective adsorption occurred, with kaolin the collagenase was destroyed.

Collagenase is not injured by alcohol (distinction from trypsin). Collagen which has been treated with picric acid withstands the action of pepsin and trypsin, but is attacked by collagenase. Collagen and fibrin pretreated with formaldehyde are com­

pletely resistant to collagenase and trypsin, but fibrin, so treated, is attacked by pepsin. Glycerol extracts of spleen attack fibrin and collagen in alkaline solu­

tion and sodium carbonate extracts contain glutinase.

Glycerol and sodium carbonate extracts of pancreas act equally well on collagen and fibrin, but those of liver are inactive. Pure pancreatic juice without addition of enterokinase digests fibrin both in acid and alkaline solution, but acts only faintly on collagen.

In presence of enterokinase, collagen is attacked in both acid and alkaline solution, but fibrin only in alkaline solution. Enterokinase alone possesses to a slight extent the properties of collagenase and glutinase. Glycerol extracts of the faeces of various animals were shown to contain tryptase, collagenase,

<x-glutinase (glutinises in acid solution), and p-glutin- ase (glutinises in alkaline solution). Diphtheria toxin contains a-glutinase, but no collagenase, peptase, or tryptase. P. W. Cl u t t e r b u c k.

G lycerop h osp h atase. H. Ko b a y a s h i (J. Bio­

chem. [Japan], 1926, 6, 261—274).—The optimum acidity for the activity of glycerophosphatase is at p a 5-56. The rate of hydrolysis of glycerophosphate is proportional to the enzyme quantity, i.e., the time necessary for equal degrees of hydrolysis is

inversely proportional to the amount of enzyme.

The affinity between the enzyme and the substrate is not influenced by the acidity of the medium.

Ch e m ic a l Ab s t r a c t s. P e r o x y d a se . IV. In fluence of th e su b str a te on th e o p tim u m ;>„. H. Uc k o and H. W. Ba n s i

(Z. physiol. Chem., 1927, 164, 52—57).—Horse­

radish peroxydase has different optimum reactions for the substrates, pyrogallol, guaiacol, and o-cresol (cf. A ., 1926, 1275). The activity-^,, curve and the optimum p n are not specific properties of an enzyme, but depend on the substrate.

A . WORMALL.

S ter eo ch em ic a l sp ecificity of lip a se s. E ffect of p o iso n s on fa t-sp littin g en zy m es. P. Ro n a

and R. Am m o n (Biochem. Z., 1927, 181, 49—79).—

The course of hydrolysis of various esters by different preparations of lipase was followed electrometrically, periodic measured additions of alkali being made in sufficient amount to maintain a constant p u ; in all cases the course of reaction was linear. When the substrate was r-methyl mandelate, the lipases from pig- and ox-liver and from taka-diastase attacked the cZ-ester preferentially, w'hilst those from human liver, guinea-pig serum, and pig-pancreas attacked the Z-ester first (cf. Willstatter, A ., 1924, i, 1144).

The lipase from pig-liver behaved abnormally in that it hydrolysed the pure Z-ester more rapidly than the pure ¿-ester, although its action on the r-ester was to bring about a preferential hydrolysis of the cZ-form; on further purification, this enzyme tended to lose its optical specificity. Whilst the concentration of enzyme had little effect on the hydrolysis of tributyrin by pig-liver lipase, it had a large effect on the rate of hydrolysis of the mandelic esters; further, methyl mandelate was hydrolysed much more rapidly than ethyl mandelate. The inhibiting effect of various alkaloids, of atoxyl, of trypafiavine, and of adrenaline on these enzymes is described. C. R . Ha r i n g t o n.

In flu en ce of th e q u in in e g ro u p o n e n z y m ic fvuictions of th e o r g a n ism . V I. D ep en d en ce on th e 7>h of th e a ction of q u in in e an d ca rb a m id e on p a n c rea tic lip a se . J. A . Sm o r o d i n c e v and V. A . Da n il o v (Biochem. Z., 1927, 181, 149— 157).

— Quinine accelerates the hydrolysis of triacetin by pancreatic lipase over the range p u 4-5— 6-0, but retards it at p n 8, which, under normal conditions, is the optimum reaction for this enzyme. Carb­

amide has no effect on the enzyme at any reaction;

carbamide nitrate has no action beyond that due to its effect on the p a of the solution.

C. R. Ha r i n g t o n. P a p a in . H. Kr a u t and E. Ba u e r (Z. physiol.

Chem., 1927, 164, 10—36; cf. A ., 1924, i, 467;

1925, i, 739).—The adsorption curves of papain and invertase on alumina show significant differences.

Small amounts of the adsorbent remove a relatively large amount of papain and a small amount of invertase, but with further additions the amount of the former decreases, whilst th a t of the latter in­

creases. Simple peptides are not integral constituents of papain, but it is not possible to separate the peptones and polypeptides from the enzyme.

Crude papain contains an inhibitory substance, which is probably a peptide, since digestion with erepsin leads to an increase in activity. Substances associated with the enzyme act as stabilisers and as co-adsorbents, for purified papain is much less readily adsorbed than the original preparation. Purification results in the removal of nine-tenths of the original impurities, and the methods of purification include extraction with chloroform, precipitation of part of the impurities with lead phosphate, adsorption on alumina, and elution with dilute acetic acid. Yeast- gum is added as a stabiliser and .co-adsorbent. D i­

alysis causes a loss in activity, probably due to the loss of a specific stabiliser with buffer properties.

Some chemical properties of the preparation are given. Millon’s reaction becomes somewhat stronger during the course of purification, whilst Molisch’s and the ninhydrin reactions remain fairly constant.

A study of the stability at different liydrogen-ion concentrations shows that there is a maximum stability at p a 5—6, approximately the same as the optimum reaction. A. Wo r m a l l.

Influence of th e reaction on th e p roteo ly tic p ow er of papain. II. W. E. Ri n g e r and B. W.

Gr u t t e r in k (Z. physiol. Chem., 1927, 164, 112—

142; cf. A., 1926, 977).—A purified water-soluble papain shows two optimum reactions (pa 2-5 and 11-3) for the dissolution of fibrin. Similar optima are found when lieat-coagulated egg-white and serum proteins are used as substrates, although the optimum on the acid side varies somewhat with the protein.

The influence of electrolytes on this action depends on the p a of the medium, for small quantities of a large number of anions and cations have a strong inhibitory action, except in neutral or faintly acid solutions, where they have a strong activating action.

Variations in the amount of electrolyte, with constant p a, produce alterations in the velocity of the reactions, and also in the range of p a, over which the action takes place. The further digestion of serum proteins by papain in neutral solution, as measured by the liberation of amino-nitrogen, is also accelerated by electrolytes. Papain and serum proteins appear to form compounds in neutral solution. A. Wo r m a l l.

Purification and p rop erties of p ep sin . J. C.

Fo r b e s (J. Biol. Chem., 1927, 71, 559—585).— On treatment of an aqueous solution of commercial pepsin with safranine, the enzyme was quantitatively precipitated; the precipitate was dissolved in 2 0% alcohol containing a little oxalic acid and the dye removed by extraction with butyl alcohol. In this way, enzyme preparations were obtained having twenty times the activity of the original material.

Such an active preparation had C 45-24, H 6-19, N 11-50, S 1-79%; figures are given for the nitrogen distribution; traces only of purine derivatives were found. The isoelectric point was at p a 2-5; the preparation had no immediate effect when added to dilute hydrochloric acid, but on prolonged incubation of such a solution a decrease was observed both in the conductivity and the hydrogen-ion concentration.

Extraction of the dried preparation with ether left the enzymic activity unchanged; addition of alcohol and ether to aqueous solutions of the pepsin caused

inactivation when the concentration of acid was high, but had little or no effect when it was low.

C. R. Ha r i n g t o n. In ten sificatio n of en zy m ic a ctio n b y m in im a l a m o u n ts of k n o w n su b sta n ce s. M. Ja c o b y (Bio- chem. Z., 1927, 181, 194—206).—The activity of urease is increased by the presence of as little as 0 -0 0 0 1 mg. of potassium cyanide, or by even less of acetaldehydecyanohydrin; the activating effect is independent of the p a. Urease which has been inactivated by mercuric chloride can be reactivated by treatment with cyanide. The action of urease is also accelerated by very small amounts of histidine, histamine, and haemoglobin; arginine and lysine have a slight accelerating effect, whilst other amino- acids have practically no effect, Histidine, therefore, apparently belongs to a second group of accelerating substances of the same order of activity as cyanides.

C. R. Ha r i n g t o n. F er m en ta tio n s w ith y e a s t re g a rd ed fr o m th e b io lo g ic a l p o in t of v iew . II. F e r m e n ta b ility of glycera ld eh y d e an d d ih y d rox ya ceton e b y liv in g y ea st. H . H a e h n and M. G l a t j b i t z (Ber., 1927, 60, [5], 490—493).—Pure glyceraldehyde is not fermented by bottom beer 3reast, top yeast M , or Saccliaromycodes L udw igii in 1% solution or in a solution of p n 7-37—7-73, and only slightly if boiled previous to the addition of the yeast. The yeast- cells become damaged to some extent. Pure dihydr­

oxyacetone is vigorously fermented by S . L udw igii after a pronounced period of induction, but not by the other two varieties of yeast. W ith technical, bimolecular dihydroxyacetone, the action is some­

what less vigorous. The yeast-cell does not suffer damage in any case. H . W r e n .

F er m en ta b ility of free an d p h o sp h o ry la ted h e x o s e s and a p o la r im e tr ic p roof of th e ir fix ­ ation in th e y e a st-c e ll. C. Nettbergand M. Ko b e l

(Biochem. Z., 1926, 179, 451— 458).— Robison’s hexosemonophosphate and the two monophosphates prepared by Neuberg—one by partial hydrolysis of hexosediphosphate, the other by hydrolysis of synthetic sucrosemonophosphate—are all fermented by top and bottom yeasts, but more rapidly by yeast juice. In no case is the fermentation as rapid as that of dextrose and laavulose. Laevulose shows an increased lsevorotation, and hexosemonophosphate (Robison’s and that from the diphosphate) an in ­ creased dextrorotation when they are added to yeast juice which has lost its fermentative power through keeping. I t is suggested that a combination of sugar or sugar phosphate with yeast protein or protein degradation products may lie at the basis of this phenomenon (see also Neuberg and Kobel, A., 1926, 151, 1061). & J . Pr y d e.

P h osp h ory la tio n an d o x id o -red u ctio n [in fer­

m entation], R. Ni l s s o n and T. Lo v g r e n (Z.

physiol. Chem., 1927, 164, 61—68).—The addition of small quantities of methylene-blue to dried bottom yeast H in a phosphate solution containing sugar decreases considerably the induction of the ferment­

ation process as measured by the evolution of carbon dioxide, but subsequently the fermentation curves, with and without methylene-blue, are approximately

BIOCHEMISTRY. 379 parallel. Large quantities of methylene-blue, how­

ever-, entirely suppress the evolution of carbon dioxide.

The action of small amounts of methylene-blue, by which the induction period is shortened, is attributed to the participation of this substance in the oxido- reduction reactions. Addition of methylgne-blue also increases the rate of disappearance of inorganic phosphate from the solution during the first stages.

A. Wormall.