Natural evidence for chemical and early biological evolution

Meteorites, particularly type II carbonaceous chondrites, provide natural, tangible evidence for chemical evolution, but they do not appear to contain any evidence for biological evolution. On the other hand, some of the oldest sedimentary rocks of the earth have yielded good evidence for early biological evolution; whatever evidence there may be for chemical evolution in these old rocks is generally obscure. Carbonaceous chondrites (types I, II, and III) have been examined for thier content of various kinds of organic compounds. Amino acids have been reported to be present in the three types, but only in type II carbonaceous chondrites (Murray and Murchison) has an indigenous suite of amino acids been found which is apparently free of most terrestrial contaminations. These indigenous compounds are thought to have resulted from extraterrestrial, abiotic, chemical syntheses, and the presence of the amino acids in meteorites provides strong support for the theory of chemical evolution. The geological record of the Swaziland Sequence and Bulawayan System of Southern Africa contains morphological and chemical fossils which indicate that early biological evolution was taking place at least 3.0 to 3.3 aeons ago. Interpretation of the significance of the chemical fossil record has proven to be difficult. At present the occurrence of simple compounds in these very ancient rocks is believed to have little or nothing to do with biochemical processes three aeons ago. The bulk of the reduced carbonaceous material in these rocks, however, probably represents the residue of three billion years old and older organic matter. Isotopic studies of this carbonaceous material may provide chemical evidence for early biological evolution.     

Regulation of hepatic l-serine dehydratase and l-serine-pyruvate aminotransferase in the developing neonatal rat

1. The activities of l-serine dehydratase and l-serine–pyruvate aminotransferase were determined in rat liver during foetal and neonatal development. 2. l-Serine–pyruvate aminotransferase activity begins to develop in late-foetal liver, increases rapidly at birth to a peak during suckling and then decreases at weaning to the adult value. 3. l-Serine dehydratase activity is very low prenatally, but increases rapidly after birth to a transient peak. After a second transient peak around the time weaning begins, activity gradually rises to the adult value. Both of these peaks have similar isoenzyme compositions. 4. In foetal liver both l-serine dehydratase and l-serine–pyruvate aminotransferase activities are increased after injection in utero of glucagon or dibutyryl cyclic AMP. Cycloheximide or actinomycin D inhibited the prenatal induction of both enzymes and actinomycin D blocked the natural increase of l-serine dehydratase immediately after birth. Glucose or insulin administration also blocked the perinatal increase of l-serine dehydratase. 5. After the first perinatal peak of l-serine dehydratase, activity is increased by cortisol and this is inhibited by actinomycin D. After the second postnatal peak, activity is increased by amino acids or cortisol and this is insensitive to actinomycin D inhibition. Glucose administration blocks the cortisol-stimulated increase in l-serine dehydratase and also partially lowers the second postnatal peak of activity. 6. The developmental patterns of the enzymes are discussed in relation to the pathways of gluconeogenesis from l-serine. The regulation of enzyme activity by hormonal and dietary factors is discussed with reference to the changes in stimuli that occur during neonatal development and to their possible mechanisms of action.     

Pig liver pyruvate carboxylase. Purification, properties and cation specificity

1. Pyruvate carboxylase was purified to apparent homogeneity from pig liver mitochondria and shown to be free of all kinetically contaminating enzymes. 2. The enzyme has a mol. wt. of 520000 and is composed of four subunits, each with a mol. wt. of 130000. 3. The enzyme can exist as the active tetramer, dimer and monomer, although the tetramer appears to be the form in which the enzyme is normally assayed. 4. For every 520000g of the enzyme there are 4mol of biotin, 3mol of zinc and 1mol of magnesium. No significant concentrations of manganese were detected. 5. Analysis by sodium dodecyl sulphate-polyacrylamide gel electrophoresis indicates three polypeptide chains per monomer unit, each with a mol. wt. of 47000. 6. The amino acid analysis, stoicheiometry of the reaction and the activity of the enzyme as a function of pH are also presented. 7. The enzyme is activated by a variety of univalent cations but not by Tris(+) or triethanolamine(+). 8. The activity of the enzyme is dependent on the presence of acetyl-CoA; the low rate in the absence of added acetyl-CoA is not due to an enzyme-bound acyl-CoA. The dissociation constant for enzyme-bound acetyl-CoA is a marked function of pH.     

Kinetics of the hydrolysis of N-benzoyl-l-serine methyl ester catalysed by bromelain and by papain. Analysis of modifier mechanisms by lattice nomography, computational methods of parameter evaluation for substrate-activated catalyses and consequences of postulated non-productive binding in bromelain- and papain-catalysed hydrolyses

1. N-Benzoyl-l-serine methyl ester was synthesized and evaluated as a substrate for bromelain (EC 3.4.22.4) and for papain (EC 3.4.22.2). 2. For the bromelain-catalysed hydrolysis at pH7.0, plots of [S(0)]/v(i) (initial substrate concn./initial velocity) versus [S(0)] are markedly curved, concave downwards. 3. Analysis by lattice nomography of a modifier kinetic mechanism in which the modifier is substrate reveals that concave-down [S(0)]/v(i) versus [S(0)] plots can arise when the ratio of the rate constants that characterize the breakdown of the binary (ES) and ternary (SES) complexes is either less than or greater than 1. In the latter case, there are severe restrictions on the values that may be taken by the ratio of the dissociation constants of the productive and non-productive binary complexes. 4. Concave-down [S(0)]/v(i) versus [S(0)] plots cannot arise from compulsory substrate activation. 5. Computational methods, based on function minimization, for determination of the apparent parameters that characterize a non-compulsory substrate-activated catalysis are described. 6. In an attempt to interpret the catalysis by bromelain of the hydrolysis of N-benzoyl-l-serine methyl ester in terms of substrate activation, the general substrate-activation model was simplified to one in which only one binary ES complex (that which gives rise directly to products) can form. 7. In terms of this model, the bromelain-catalysed hydrolysis of N-benzoyl-l-serine methyl ester at pH7.0, I=0.1 and 25 degrees C is characterized by K(m) (1) (the dissociation constant of ES)=1.22+/-0.73mm, k (the rate constant for the breakdown of ES to E+products, P)=1.57x10(-2)+/-0.32x10(-2)s(-1), K(a) (2) (the dissociation constant that characterizes the breakdown of SES to ES and S)=0.38+/-0.06m, and k' (the rate constant for the breakdown of SES to E+P+S)=0.45+/-0.04s(-1). 8. These parameters are compared with those in the literature that characterize the bromelain-catalysed hydrolysis of alpha-N-benzoyl-l-arginine ethyl ester and of alpha-N-benzoyl-l-arginine amide; K(m) (1) and k for the serine ester hydrolysis are somewhat similar to K(m) and k(cat.) for the arginine amide hydrolysis and K(as) and k' for the serine ester hydrolysis are somewhat similar to K(m) and k(cat.) for the arginine ester hydrolysis. 9. A previous interpretation of the inter-relationships of the values of k(cat.) and K(m) for the bromelain-catalysed hydrolysis of the arginine ester and amide substrates is discussed critically and an alternative interpretation involving substantial non-productive binding of the arginine amide substrate to bromelain is suggested. 10. The parameters for the bromelain-catalysed hydrolysis of the serine ester substrate are tentatively interpreted in terms of non-productive binding in the binary complex and a decrease of this type of binding by ternary complex-formation. 11. The Michaelis parameters for the papain-catalysed hydrolysis of the serine ester substrate (K(m)=52+/-4mm, k(cat.)=2.80+/-0.1s(-1) at pH7.0, I=0.1, 25.0 degrees C) are similar to those for the papain-catalysed hydrolysis of methyl hippurate. 12. Urea and guanidine hydrochloride at concentrations of 1m have only small effects on the kinetic parameters for the hydrolysis of the serine ester substrate catalysed by bromelain and by papain.     

Pig liver pyruvate carboxylase. The reaction pathway for the carboxylation of pyruvate

1. The reaction pathway for the carboxylation of pyruvate, catalysed by pig liver pyruvate carboxylase, was studied in the presence of saturating concentrations of K(+) and acetyl-CoA. 2. Free Mg(2+) binds to the enzyme in an equilibrium fashion and remains bound during all further catalytic cycles. MgATP(2-) binds next, followed by HCO(3) (-) and then pyruvate. Oxaloacetate is released before the random release, at equilibrium, of P(i) and MgADP(-). 3. This reaction pathway is compared with the double displacement (Ping Pong) mechanisms that have previously been described for pyruvate carboxylases from other sources. The reaction pathway proposed for the pig liver enzyme is superior in that it shows no kinetic inconsistencies and satisfactorily explains the low rate of the ATP[unk][(32)P]P(i) equilibrium exchange reaction. 4. Values are presented for the stability constants of the magnesium complexes of ATP, ADP, acetyl-CoA, P(i), pyruvate and oxaloacetate.     

Root Formation by Detached White Mustard (Sinapis alba) Cotyledons

Rooting is shown to occur in excised cotyledons of Sinapis alba when grown in petri dishes on moist filter paper. Cotyledons were excised at intervals from 6 hours after the start of imbibition, when they were yellow, unexpanded and enclosed within the testa, to 27 days after sowing when the cotyledons were green and fully expanded and on plants possessing up to 3 foliage leaves. Rooting generally began 5 or 6 days after excision and was completed dining the following 5 days. The age of cotyledons at t ho time of excision had three effects on rooting: the lag period be-fore rooting began and the period during which rooting took place both increased with age: but the most marked effect was on the total number of cotyledons which were able to form roots, which increased until cotyledon expansion was almost complete, then decreased as the mature cotyledons became older. Optimal rooting was shown by cotyledons detached 8 days alter sowing, when they were half expanded. At this age S5 % of them formed roots between 6 days and 8 days after excision.     

Clinical Trial of Lincomycin Hydrochloride in Reiter's Disease

A double-blind trial comparing lincomycin hydrochloride (Mycivin) and placebo in 22 patients with Reiter''s disease showed no significant difference in clinical or laboratory findings between the two groups. It is concluded that lincomycin hydrochloride is no more effective than placebo in the treatment of Reiter''s disease.     

Natural History of Isolated Patent Ductus Arteriosus and the Effect of Surgical Correction: Twenty Years' Experience at The Hospital for Sick Children,Toronto

Seven hundred and forty-four cases of patent ductus arteriosus were diagnosed at The Hospital for Sick Children, Toronto, in the 20-year period from 1947 to 1966. Of these 705 have been treated surgically. It is concluded that operation is best performed between 6 and 12 months of age. Isolated ductus arteriosus is rarely a cause of congestive heart failure, and is more likely to be so in the presence of the rubella syndrome. Failure associated with the anomaly in the neonatal period can usually be successfully managed by medical treatment. Spontaneous closure after one month of age is not to be expected. Surgical treatment at the appropriate time prevents the subsequent development of Eisenmenger''s complex.