INFLUENCE OF A SLIGHT MODIFICATION OF THE COLLODION MEMBRANE ON THE SIGN OF THE ELECTRIFICATION OF WATER

1. It is shown that collodion membranes which have received one treatment with a 1 per cent gelatin solution show for a long time (if not permanently) afterwards a different osmotic behavior from collodion membranes not treated with gelatin. This difference shows itself only towards solutions of those electrolytes which have a tendency to induce a negative electrification of the water particles diffusing through the membrane, namely solutions of acids, acid salts, and of salts with trivalent and tetravalent cations; while the osmotic behavior of the two types of membranes towards solutions of salts and alkalies, which induce a positive electrification of the water particles diffusing through the membrane, is the same. 2. When we separate solutions of salts with trivalent cation, e.g. LaCl₃ or AlCl₃, from pure water by a collodion membrane treated with gelatin, water diffuses rapidly into the solution; while no water diffuses into the solution when the collodion membrane has received no gelatin treatment. 3. When we separate solutions of acid from pure water by a membrane previously treated with gelatin, negative osmosis occurs; i.e., practically no water can diffuse into the solution, while the molecules of solution and some water diffuse out. When we separate solutions of acid from pure water by collodion membranes not treated with gelatin, positive osmosis will occur; i.e., water will diffuse rapidly into the solution and the more rapidly the higher the valency of the anion. 4. These differences occur only in that range of concentrations of electrolytes inside of which the forces determining the rate of diffusion of water through the membrane are predominantly electrical; i.e., in concentrations from 0 to about M/16. For higher concentrations of the same electrolytes, where the forces determining the rate of diffusion are molecular, the osmotic behavior of the two types of membranes is essentially the same. 5. The differences in the osmotic behavior of the two types of membranes are not due to differences in the permeability of the membranes for solutes since it is shown that acids diffuse with the same rate through both kinds of membranes. 6. It is shown that the differences in the osmotic behavior of the two types of collodion membranes towards solutions of acids and of salts with trivalent cation are due to the fact that in the presence of these electrolytes water diffuses in the form of negatively charged particles through the membranes previously treated with gelatin, and in the form of positively charged particles through collodion membranes not treated with gelatin. 7. A treatment of the collodion membranes with casein, egg albumin, blood albumin, or edestin affects the behavior of the membrane towards salts with trivalent or tetravalent cations and towards acids in the same way as does a treatment with gelatin; while a treatment of the membranes with peptone prepared from egg albumin, with alanine, or with starch has no such effect.     

THE RELATION BETWEEN THE ELECTRICAL CONDUCTIVITY OF THE EXTERNAL MEDIUM AND THE RATE OF CELL DIVISION IN SEA URCHIN EGGS

Dilution of sea water with isotonic sugar solution leaves the rate of cleavage of Arbacia eggs almost unchanged until the proportion of sea water is decreased to 20 or 25 volumes per cent. From this point cleavage becomes progressively slower with further dilution. Many eggs fail to cleave at dilutions of 5 to 6 volumes per cent. No cleavage occurs in 2 volumes per cent sea water or in pure sugar solution. Eggs returned from these media to sea water resume cleavage and development. There is thus no relation between the rate of cleavage and the electrical conductivity of the medium, except possibly within the range of dilutions from 20 to 5 volumes per cent sea water. In this range cleavage rate decreases as conductivity decreases, but the relation is not a linear one.     

ON THE GROWTH AND RESPIRATION OF SULFUR-OXIDIZING BACTERIA

1. It is shown that Sulfomonas thiooxidans oxidizes elementary sulfur completely to sulfuric acid. Sodium thiosulfate is oxidized by this organism completely to sulfate. Sulfomonas thiooxidans differs, in this respect, from various other sulfur-oxidizing bacilli which either produce elementary sulfur, from the thiosulfate, or convert it into sulfates and persulfates. 2. The organism derives its carbon from the CO₂ of the atmosphere, but is incapable of deriving the carbon from carbonates or organic matter. 3. The S:C, or ratio between the amount of sulfur oxidized to sulfate and amount of carbon assimilated chemosynthetically from the CO₂ of the atmosphere, is, with elementary sulfur as a source of energy, 31.8, and with thiosulfate 64.2. The higher ratio in the case of the thiosulfate is due to the smaller amount of energy liberated in the oxidation of sulfur compound than in the elementary form. 4. Of the total energy made available in the oxidation of the sulfur to sulfuric acid, only 6.65 per cent is used by the organism for the reduction of atmospheric CO₂ and assimilation of carbon. 5. Sulfates do not exert any injurious effect upon sulfur oxidation by Sulfomonas thiooxidans. Any effect obtained is due to the cation rather than the sulfate radical. Nitrates exert a distinctly injurious action both on the growth and respiration of the organism. 6. There is a definite correlation between the amount of sulfur present and velocity of oxidation, very similar to that found in the growth of yeasts and nitrifying bacteria. Oxidation reaches a maximum with about 25 gm. of sulfur added to 100 cc. of medium. However, larger amounts of sulfur have no injurious effect. 7. Dextrose does not exert any appreciable injurious effect in concentrations less than 5 per cent. The injurious effect of peptone sets in at 0.1 per cent concentration and brings sulfur oxidation almost to a standstill in 1 per cent concentration. Dextrose does not exert any appreciable influence upon sulfur oxidation and carbon assimilation from the carbon dioxide of the atmosphere. 8. Sulfomonas thiooxidans can withstand large concentrations of sulfuric acid. The oxidation of sulfur is affected only to a small extent even by 0.25 molar initial concentration of the acid. In 0.5 molar solutions, the injurious effect becomes marked. The organism may produce as much as 1.5 molar acid, without being destroyed. 9. Growth is at an optimum at a hydrogen ion concentration equivalent to pH 2.0 to 5.5, dropping down rapidly on the alkaline side, but not to such an extent on the acid, particularly when a pure culture is employed. 10. Respiration of the sulfur-oxidizing bacteria can be studied by using the filtrate of a vigorously growing culture, to which a definite amount of sulfur is added, and incubating for 12 to 24 hours.     

STUDIES ON THE CROSS-STRIATION OF THE INDIRECT FLIGHT MYOFIBRILS OF THE BLOWFLY CALLIPHORA

1. The cross-striation in the indirect flight myofibrils of Calliphora has been studied by phase contrast and polarised light microscopy. The band pattern at rest-length has been determined in flies killed in osmium tetroxide vapour while their wings remained in the resting position. All other observations have been made on unfixed fibrils. Although length changes in situ are probably very slight (about 2 per cent), isolated fibrils, by treatment with crude muscle extract or with ATP, can be induced to elongate to 104 per cent rest-length, or to shorten by 8 per cent but no more. Over the range 98 to 104 per cent rest-length, experimentally induced length changes are reversible. The fibrils can also be stretched beyond 104 per cent rest-length, but the process is irreversible. During the course of glycerol extraction the fibrils elongate to 104 per cent rest-length. 2. The changes in band pattern observed over the range 104 to 92 per cent rest-length are qualitatively the same as the changes observed over a wider range (about 130 to 40 per cent rest-length) in the skeletal myofibrils of rabbits. The earlier stages of shortening appear to be effected by retraction of the I bands into the A bands where they fill up the H zones. No evidence has been found that any changes in band pattern are due to a migration of the A substance. 3. Two components of the sarcomere can be extracted from it and a third component remains behind. These three components, which have also been demonstrated in skeletal myofibrils of the rabbit, where they behave in the same way, are: (a) the A substance which does not change its position as the fibril changes its length, and which can be extracted by the same procedures as remove myosin (shown elsewhere to be the A substance) from rabbit fibrils; (b) a material which extends from the Z lines to the borders of the H zone and which moves inwards during contraction and outwards during elongation; it can capture rabbit myosin from solution and form with it a contractile system, and it is thought to be actin; (c) a "backbone" or stroma bearing Z and M lines. 4. Since all these features of the cross-striation are the same in the insect fibrils as in rabbit fibrils, it is considered very probable that the sarcomere is similarly organised in both types of muscle and contracts by essentially the same mechanism.     

TIME RELATIONS OF GROWTH : III. GROWTH CONSTANTS DURING THE SELF-ACCELERATING PHASE OF GROWTH.

Growth curves consist, in all cases, of two major segments. The first major segment is, in the case of higher animals and plants, made up in turn of several (probably five) shorter segments during each of which growth takes place at a constant percentage rate. The transitions between the successive stages are abrupt, the abruptness being of the order of metamorphosis in cold blooded animals. It has been made clear in the first paper of this series that the time rate of growth following the major inflection declines at a constant percentage rate. The junction between the two major segments occurs at puberty in animals and flowering in plants. The two major segments are not symmetrical about the major inflection. The slope of the segment following the inflection is always less than the slope of the segment preceding the inflection. The major inflection does not occur in the center of the growth curve. The instantaneous rate of growth at the beginning of growth is of the order of 100–200 per cent per day (i.e. the body weight is doubled in from 7 to 17 hours). It may be mentioned that 2 months after conception the rate of growth in man is only 8 per cent per day. This is contrary to all the published statements. Thus, Minot concluded that growth begins at 1000 per cent per day; Jackson concluded that in man, growth during the 1st month takes place at 57.5 million per cent per month; during the 2nd month 990 per cent per month; during the 3rd month 390 per cent per month (8 per cent per day is only 240 per cent per month). The reason for the discrepancy between the values derived, by the method adopted by the writer, and the values given in the literature is explained by Fig. 1.     

THE EFFECT OF CARBON DIOXIDE TENSION ON THE METABOLISM OF CEREBRAL CORTEX AND MEDULLA OBLONGATA

Manometric measurements were made of oxygen uptake (Q _OO2) and aerobic lactic acid output (Q_G) by slices of cerebral cortex and medulla oblongata of the cat in the presence of mixtures of 1, 5, and 20 volumes per cent of carbon dioxide in oxygen. The concentrations of NaHCO₃ and NaCl in the medium were varied to maintain constant pH and sodium ion concentrations. The calcium ion concentration was 0.0002 M. At pH 7.5 under these conditions, an increase in carbon dioxide from 1 per cent to 5 per cent doubled the Q_G of both tissues but did not alter Q _OO2; an increase from 5 per cent to 20 per cent carbon dioxide had no further effect on Q_G in either tissue or Q _OO2 of cortex, but did depress the Q _OO2 of medulla. At pH 8.1, an increase in carbon dioxide from 1 per cent to 5 per cent raised the Q _OO2 and Q_G of cortex by about 60 per cent. Measurements at low oxygen tension carried out previously in phosphate medium were repeated in bicarbonate medium to obtain data for the combined output of lactic acid and carbon dioxide (Q_A). When the oxygen in the gas phase was decreased from 95 to 3 volumes per cent, the lactic acid output as measured colorimetrically increased by 114 mg./gm. in cortex and by 8 mg./gm. in medulla; Q_A increased from 12.3 to 13.5 in cortex and decreased from 5.1 to 3.8 in medulla.     

THE CHLOROPHYLL-PROTEIN COMPOUND OF THE GREEN LEAF

1. Aqueous extracts of spinach and Aspidistra leaves yield highly opalescent preparations which are not in true solution. Such extracts differ markedly from colloidal chlorophyll in their spectrum and fluorescence. The differences between the green leaf pigment and chlorophyll in organic solvents are shown to be due to combination of chlorophyll with protein in the leaf. 2. The effect of some agents on extracts of the chlorophyll-protein compound has been investigated. Both strong acid and alkali modify the absorption spectrum, acid converting the compound to the phaeophytin derivative and alkali saponifying the esterified groups of chlorophyll. Even weakly acid solutions (pH 4.5) denature the protein. Heating denatures the protein and modifies the absorption spectrum and fluorescence as earlier described for the intact leaf. The protein is denatured by drying. Low concentrations of alcohol or acetone precipitate and denature the protein; higher concentrations cause dissociation liberating the pigments. 3. Detergents such as digitonin, bile salts, and sodium desoxycholate clarify the leaf extracts but denature the protein changing the spectrum and other properties. 4. Inhibiting agents of photosynthesis are without effect on the absorption spectrum of the chlorophyll-protein compound. 5. The red absorption band of chlorophyll possesses the same extinction value in organic solvents such as ether or petroleum ether, and in aqueous leaf extracts clarified by digitonin although the band positions are different. Using previously determined values of the extinction coefficients of purified chlorophylls a and b, the chlorophyll content of the leaf extracts may be estimated spectrophotometrically. 6. It was found that the average chlorophyll content of the purified chloroplasts was 7.86 per cent. The protein content was 46.5 per cent yielding an average value of 16.1 parts per 100 parts of protein. This corresponds to a chlorophyll content of three molecules of chlorophyll a and one of chlorophyll bfor the Svedberg unit of 17,500. It is suggested that this may represent a definite combining ratio of a and b in the protein molecule.     

PRODUCTS OF THE OXIDATION OF THIOSULFATE BY BACTERIA IN MINERAL MEDIA

Various cultures (previously described), which oxidize thiosulfate in mineral media have been studied in an attempt to determine the products of oxidation. The transformation of sodium thiosulfate by Cultures B, T, and K yields sodium tetrathionate and sodium hydroxide; secondary chemical reactions result in the accumulation of some tri- and pentathionates, sulfate, and elemental sulfur. As a result of the initial reaction, the pH increases; the secondary reactions cause a drop in pH after this initial rise. The primary reaction yields much less energy than the reactions effected by autotrophic bacteria. No significant amounts of assimilated organic carbon were detected in media supporting representatives of these cultures. It is concluded that they are heterotrophic bacteria. Th. novellus oxidizes sodium thiosulfate to sodium sulfate and sulfuric acid; the pH drops progressively with growth and oxidation. Carbon assimilation typical of autotrophic bacteria was detected; the ratio of sulfate-sulfur formed to carbon assimilated was 56:1. It is calculated that 5.1 per cent of the energy yielded by the oxidation of thiosulfate is accounted for in the organic cell substance synthesized from inorganic materials. This organism is a facultative autotroph. The products of oxidation of sodium thiosulfate by Th. thioparus are sodium sulfate, sulfuric acid, and elemental sulfur; the ratio of sulfate sulfur to elemental sulfur is 3 to 2. The pH decreases during growth and oxidation. The elemental sulfur is produced by the primary reaction and is not a product of secondary chemical changes. The bacterium synthesizes organic compounds from mineral substances during growth. The ratio of thiosulfate-sulfur oxidized to carbon assimilated was 125:1, with 4.7 per cent of the energy of oxidation recovered as organic cell substance. This bacterium is a strict autotroph.     

THE TOTAL LUMINOUS EFFICIENCY OF LUMINOUS BACTERIA

Methods are described for measuring the light emitted by an emulsion of luminous bacteria of given thickness, and calculating the light emitted by a single bacterium, measuring 1.1 x 2.2 micra, provided there is no absorption of light in the emulsion. At the same time, the oxygen consumed by a single bacterium was measured by recording the time for the bacteria to use up .9 of the oxygen dissolved in sea water from air (20 per cent oxygen). The luminescence intensity does not diminish until the oxygen concentration falls below 2 per cent, when the luminescence diminishes rapidly. Above 2 per cent oxygen (when the oxygen dissolving in sea water from pure oxygen at 760 mm. Hg pressure = 100 per cent) the bacteria use equal amounts of oxygen in equal times, while below 2 per cent oxygen it seems very likely that rate of oxygen absorption is proportional to oxygen concentration. By measuring the time for a tube of luminous bacteria of known concentration saturated with air (20 per cent oxygen) to begin to darken (2 per cent oxygen) we can calculate the oxygen absorbed by one bacterium per second. The bacteria per cc. are counted on a blood counting slide or by a centrifugal method, after measuring the volume of a single bacterium (1.695 x 10^–12 cc.). Both methods gave results in good agreement with each other. The maximum value for the light from a single bacterium was 24 x 10^–14 lumens or 1.9 x 10^–14 candles. The maximum value for lumen-seconds per mg. of oxygen absorbed was 14. The average value for lumen-seconds per mg. O₂ was 9.25. The maximum values were selected in calculating the efficiency of light production, since some of the bacteria counted may not be producing light, although they may still be using oxygen. The "diet" of the bacteria was 60 per cent glycerol and 40 per cent peptone. To oxidize this mixture each mg. of oxygen would yield 3.38 gm. calories or 14.1 watts per second. 1 lumen per watt is therefore produced by a normal bacterium which emits 14 lumen-seconds per mg. O₂ absorbed. Since the maximum lumens per watt are 640, representing 100 per cent efficiency, the total luminous efficiency if .00156. As some of the oxygen is used in respiratory oxidation which may have nothing to do with luminescence, the luminescence efficiency must be higher than 1 lumen per watt. Experiments with KCN show that this substance may reduce the oxygen consumption to 1/20 of its former value while reducing the luminescence intensity only ¼. A partial separation of respiratory from luminescence oxidations is therefore effected by KCN, and our efficiency becomes 5 lumens per watt, or .0078. This is an over-all efficiency, based on the energy value of the "fuel" of the bacteria, regarded as a power plant for producing light. It compares very favorably with the 1.6 lumens per watt of a tungsten vacuum lamp or the 3.9 lumens per watt of a tungsten nitrogen lamp, if we correct the usual values for these illuminants, based on watts at the lamp terminals, for a 20 per cent efficiency of the power plant converting the energy of coal fuel into electric current. The specific luminous emission of the bacteria is 3.14 x 10^–6 lumens per cm². One bacterium absorbs 215,000 molecules of oxygen per second and emits 1,280 quanta of light at λ_max = 510µµ. If we suppose that a molecule of oxygen uniting with luminous material gives rise to the emission of 1 quantum of light energy, only 1/168 of the oxygen absorbed is used in luminescence. On this basis the efficiency becomes 168 lumens per watt or 26.2 per cent.     

THE BIOGENESIS OF PRIMARY SEX HORMONES : I. THE FATE OF ESTRINS INJECTED INTO THE RABBIT

1. A method is given for the extraction and fractionation of rabbit urines which frees these urines of inactive chromogens but permits a quantitative recovery of estrone and estriol for the colorimetric determination of these compounds. 2. Estrone and estriol content of rabbit urine extracts can be determined by the concentration of the colored compound they form upon diazotization with sulfanilic acid and by the modified phenolsulfonic acid test of Cohen and Marrian. Estriol can be determined by the specific reaction first described by David. The technique for these tests is presented. 3. Estriol (300 micrograms) injected into rabbits (a) in heat, (b) pregnant, (c) pseudopregnant, (d) hysterectomized in heat, (e) hysterectomized pseudopregnant, (f) ovariectomized, is excreted in the urine as estriol. Rabbit does in the luteal phase (b, c, and e) excrete 3 to 4 times the amount of estriol excreted by females without corpora lutea (a, d, and f). 4. When estrone (300 micrograms) is injected into the same types of rabbit does types a, b, and c excrete both estrone and estriol, type f excretes both estrone and estriol shortly after ovariectomy, but only estrone at 2 months after castration. Hysterectomized animals (types d and e) never excrete estriol after estrone injection. The total urinary estrin (estrone plus estriol) in estrone-injected animals is increased 2 to 3 times in animals in the luteal phase (b, c, and e). 5. It is concluded that the uterus is the site of conversion of estrone to estriol, and that the conversion cannot take place in a uterus completely free of ovarian control (e.g., in long time ovariectomized animals). 6. In neither estrone-injected nor estriol-injected females is all the injected hormone recovered in the urine. The maximum recovery is 66 per cent. When estrone-benzoate (600 micrograms) is injected 94–98 per cent of the hormone is recovered from animals in the luteal phase (types c and e) and about 79 per cent in an ovariectomized female (type f). These data are taken to indicate that luteal secretions give partial protection against destruction to the hormones. 7. The observation that in certain of the urine extracts the hormone titer by bioassay is somewhat higher than the colorimetric titer may indicate that there is a slight conversion of estrone to estradiol, particularly since no equilenin was found in any of the extracts by colorimetric test. 8. The simultaneous injection of 300 micrograms of estrone and 500 micrograms of progesterone 4 days after an initial injection of 300 micrograms of estrone results in: (1) an increased estrin excretion in females in heat, hysterectomized unmated, and ovariectomized, and a slight decrease in the pseudopregnant female; (2) the appearance of estriol in the urine of the long time ovariectomized animal with no urinary estriol in a control ovariectomized animal receiving no progesterone. These findings are taken to prove that the conversion of estrone to estriol occurs in the uterus under the influence of progesterone. Since animals in heat produce small amounts of estriol after estrone injection it is inferred that the ovaries of estrus rabbits produce small amounts of corpus luteum hormone in the absence of formed corpora lutea.     

A comparative study of the effects of the diamine oxidase inhibitor aminoguanidine, with or without dietary restriction, on the nucleic acid and protein composition of cardiac and type I and type II skeletal muscles of the rat.

It has been proposed that the diamine oxidase inhibitor aminoguanidine may be a potential therapeutically important anabolic agent. An investigation was therefore made into the effects of aminoguanidine treatment with or without nutritional restriction, on cardiac and skeletal muscles containing mainly of either Type I (i.e. soleus) or Type II fibres (i.e. plantaris) or a mixture of Type I and II fibres (i.e. gastrocnemius). After 3 weeks, dietary restrictions reduced cardiac weight, protein, RNA and DNA contents by between 31 per cent and 36 per cent. Similar, but smaller, reductions were observed in the soleus (18-31 per cent), plantaris (22-34 per cent) and gastrocnemius (22-34 per cent). Aminoguanidine had no effect on the heart of the rats fed ad libitum, nor did it alter the response to dietary restriction. Treatment with aminoguanidine had no overt anabolic effect on skeletal muscle, but a reduction in DNA content was observed. It was concluded that cardiac protein and nucleic acid contents are more sensitive to dietary deprivation than either anaerobic or aerobic skeletal muscles. Furthermore, aminoguanidine does not appear to promote growth or reduce catabolism as previous studies have suggested.     

THE EFFECTS OF CAFFEINE ON OXYGEN CONSUMPTION AND CELL DIVISION IN THE FERTILIZED EGG OF THE SEA URCHIN,ARBACIA PUNCTULATA

1. By means of the Warburg-Barcroft microrespirometer apparatus and the Warburg direct method, the relative effect of caffeine upon the O₂ consumption of the fertilized egg of Arbacia punctulata was shown for the following concentrations in sea water: 0.002 per cent (M/10,000), 0.004 per cent (M/5,000), 0.02 per cent (M/1,000), 0.1 per cent (M/200), 0.2 per cent (M/100), 0.5 per cent (M/40), and 2 per cent (M/10). 2. In comparison with the normal eggs (uninhibited, non-caffeine-treated controls), caffeine in concentrations including and greater than 0.1 per cent (M/200) depressed the average uptake from approximately 25 to 61 per cent over the 3 hour period. In a number of instances, as typified by Experiment 10, the effective inhibitory concentration ranged from 0.02 per cent (M/1,000) upward and the degree of depression of the O₂ consumption ranged from 10.6 per cent to 60.6 per cent. 3. All caffeine concentrations including and above 0.02 per cent (M/1,000) in the series used, resulted in decreasing the normal rate of cleavage division in the fertilized Arbacia eggs. 4. The higher concentrations (0.5 and 2 per cent) produced a complete blockage of the cleavage process. 5. Complete cleavage inhibition was noted only when the O₂ uptake had been depressed to 50 per cent or more of the normal controls. 6. O₂ consumption-time relationship data indicate an average depression, in O₂ consumption over a 3 hour period, ranging from 25 per cent with a caffeine concentration of 0.1 per cent to a 61 per cent inhibition with a concentration of 2 per cent. 7. Concentrations of less than 0.1 per cent (certainly of less than 0.02 per cent) give variable results and indicate no significant effect. 8. It is inferred from the respiration data presented that it is probable that the inhibition of the O₂ consumption in fertilized Arbacia eggs is due to the influence of caffeine upon the main (activity or primary) pathway. It will be observed that there are certain similarities of the caffeine data to the degree of inhibition accomplished by sodium cyanide. Moreover, it has been demonstrated that the cyanide probably acts on the cytochrome oxidase step in the cytochrome oxidase-cytochrome chain of reactions constituting the O₂ uptake phase of respiratory metabolism. It is not improbable, therefore, that caffeine also may act upon the cytochrome oxidase enzyme. 9. From the viewpoint of environmental conditions influencing reproductive phenomena, it is of interest that caffeine can affect the normal metabolism of the zygote.     

BIOCHEMICAL EFFECTS OF THYROID DEFICIENCY ON THE DEVELOPING BRAIN

Abstract— The effects of neonatal thyroidectomy on some constituents of the cerebrum, cerebellum and liver of the rat have been studied during the first 7 weeks of life. In the normal rat between the 6th and 14th post-natal days the RNA content per unit of DNA in the brain increased by 70 per cent. Although the brain continued to grow from the 14th to the 35th day, the amount of RNA relative to DNA decreased by about 20 per cent. The ratio of protein to DNA increased during the whole period studied and in the cerebral cortex it was more than trebled between the age of 6 and 35 days. The growth of the cerebellum extended over a longer period than that of the cerebrum, its weight increasing by 88 per cent between the ages of 14 and 35 days as compared with a cerebral increase of 34 per cent. The DNA content showed a 50 per cent increase during this period. Qualitatively these maturational changes were not affected by neonatal thyroidectomy. Quantitative changes, which applied equally to the cerebral cortex and brain as a whole, were observed. At the age of 35 days, the weights of the cerebral hemispheres and cerebellum were reduced by thyroidectomy by 20 per cent; the overall DNA content per organ did not change, but the amounts of protein and RNA relative to DNA decreased significantly. It is therefore inferred that thyroid deficiency affects the size of the cells in brain and cerebellum rather than their total number. Conversely, the cell population of the liver was only a quarter of that in the control. There was a small but significant decrease in the hepatic protein and RNA content in the hypothyroid animal. The activities of the following enzymes which served as markers for subcellular fractions in homogenates of cerebral cortex were determined: lactate dehydrogenase for the supernatant, glutamate dehydrogenase for the mitochondrial and glutamate decarboxylase for the synaptosomal fractions. When the activities were expressed on a fresh weight basis a significant decrease by comparison with the control values was observed only in the case of glutamate decarboxylase (—15 per cent at the age of 17–32 days); when the activities were based on DNA content all values were reduced, probably as a result of the general decrease in cell size. Pyrimidine metabolism of brain and liver, studied after the administration of [6-¹⁴C]-orotic acid, was not affected in either tissue by neonatal thyroidectomy. A small but significant reduction in the incorporation of labelled pyrimidine nucleotides in liver RNA was observed, but no significant decrease in the incorporation in cerebral RNA was found in the hypothyroid rats.     

THE RHEOLOGY OF THE BLOOD. V

A study has been made of those proteins which might offer exceptions to the law that the fluidity of a protein solution is a linear function of the volume concentration; viz., egg albumin, serum albumin, pseudoglobulin, euglobulin, gelatin, and sodium caseinogenate. Solutions of egg albumin below 20 per cent by weight obey the above law but somewhat below 30 per cent the fluidities begin to be too high, presumably due to the contribution to the fluidity made by the deformation of the particles as they come into contact, as the fluidity approaches zero. The fluidity of serum albumin solutions shows a similar behavior, being exceptional above 15 per cent in weight. Pseudoglobulin and euglobulin give fluidity-concentration curves (Fig. 4) which are linear up to about 2.5 per cent each in a total range of 20 and 14 per cent respectively. From this singular point both compounds show a second range which is linear. Pseudoglobulin is the only substance whose solutions seem to show a third linear range. We have also used the data of Chick and Martin for sodium caseinogenate and found evidence for two linear régimes. It is desirable at this time to call attention to the measurements of the flow of glycogen solutions by Botazzi and d''Errico (14) which in Fluidity and See PDF for Structure plasticity, page 207, are expressed in rhes. The data show two linear fluidity curves of different slopes. In this case it was definitely known that the data for each curve were measured with different viscometers which suggested the possibility of an error in viscometry entering in to confuse the issue. We have no suspicions as to the reliability of the data studied in this paper; we only wish to caution the readers that our hypotheses based on these data must be regarded with due reserve until confirmed. We have found a formula (11) based on the supposed linear relation between logarithmic fluidities and concentration which is convenient to use within the range, but close examination reveals that it does not reproduce the data for the higher concentrations at 25° nor does it permit extrapolation to pure water It is not realistic enough because it does not contemplate any change of régime in going from viscous to non-Newtonian or plastic flow. The formula does not apply to any other of the proteins studied in this paper nor to the great majority of proteins already reported as following the linear law. These are serious objections. We have therefore offered as an alternative a simple formula (24) according to which the fluidities are additive in the viscous régime. When the emulsoid particles approach close packing, they are deformed and this deformation contributes to the flow and the fluidity volume concentration curve is again linear. In fact, there may be one or more additional changes of régime.     

ON THE CHEMICAL COMPOSITION OF THE AXOPLASM OF SQUID GIANT NERVE FIBERS WITH PARTICULAR REFERENCE TO ITS ION PATTERN

Investigations dealing with the determination of the major chemical constituents of the axoplasm of the giant nerve fiber of the squid are described. Particular emphasis has been placed on determining the components involved in acid-base balance. It was found that 72 per cent of the total solids of axoplasm, representing 13.5 per cent of the wet material, are of relatively low molecular weight (dialyzable) and consist mainly of charged ionic or dipolar constituents. Of the 520 micromoles per gm. of total base, 72 per cent are balanced by organic acids: aspartic acid (65 micro equivalents per gm.), glutamic acid (10 micro equivalents), fumaric and succinic acids (15 micro equivalents), a new polycarboxylic acid (35 micro equivalents), and isethionic acid, a biologically novel sulfonic acid (220 micro equivalents). Besides potassium, sodium, small amounts of calcium, and magnesium there is a considerable fraction of organic (nitrogenous) base. Other features of the chemical composition of squid axoplasm include a relatively high concentration of taurine (100 micro equivalents) and an ultraviolet absorbing substance possibly identical with N-methylpicolinic acid. The distribution of the phosphates, especially the concentration of ATP, has been investigated. Specific techniques elaborated in connection with this study have been described and the biochemical implications of the analytical results are discussed.     

THE CONDITIONS OF RECOVERY OF TRANSMISSIVITY OF NEWLY REPASSIVATED IRON WIRES IN NITRIC ACID

1. Passive steel wires were activated in a bath (Bath A) containing 70 v. per cent HNO₃ (in which they undergo prompt repassivation), and immediately transferred to a second bath (Bath B) containing HNO₃ of a concentration varying in different experiments. After varying intervals in this bath they were transferred while still passive to a third bath (Bath C) containing strong HNO₃ (70 or 100 v. per cent) and there immediately activated. 2. During the immersion in Bath B the wires progressively recover their ability to transmit activation waves in strong HNO₃. The measure of this recovery is the distance travelled by the activation waves in Bath C after the varying times of exposure in Bath B. Transmissivity as thus measured is at first incomplete (decremental) and later becomes complete. The minimal exposures in Bath B required to render wires completely transmissive in the strong acid of Bath C were determined for concentrations of HNO₃ between 10 and 100 v. per cent. With 100 v. per cent HNO₃ in Bath C, these exposures range from 40 minutes or more in 15 v. per cent to 10 minutes in 100 v. per cent HNO₃ (temperature 19–20° in all baths). 3. The time required for complete recovery varies inversely with the concentration of the acid in the recovery bath (Bath B), but increases rapidly with the concentration of the acid in the testing bath (Bath C). Hence at a time when a wire has recovered just sufficiently to transmit non-decrementally in a given strong acid (e.g., 70 v. per cent) it still transmits decrementally in a stronger acid. Complete recovery for transmission in 100 v. per cent HNO₃ requires about twice as long as for 70 v. per cent HNO₃. In HNO₃ of 50 v. per cent and less decremental transmission does not occur. 4. The indications are that recovery is an effect of the progressive solvent action of the external acid on the passivating oxide film, which at its first deposition appears to be relatively thick and hence resistant to electrochemical reduction. The final stage of recovery, when electrical sensitivity and speed of transmission are maximal, would on this hypothesis correspond to minimal thickness, possibly monomolecular. 5. The rate of recovery in Bath B is not far from proportional to the concentration of HNO₃ in the more dilute solutions, but in the higher, especially the strongly passivating, concentrations (70 to 100 v. per cent) the rate becomes appreciably slower than proportional, apparently because of the intense oxidizing action of these solutions, which reinforces the oxide sheet and retards the thinning process. 6. The bearing of these observations on the problem of the conditions of recovery in irritable living tissues (such as nerve) during the absolute and relative refractory periods is briefly discussed.     

AXONAL TRANSPORT OF RADIOACTIVITY IN THE GOLDFISH OPTIC SYSTEM FOLLOWING INTRAOCULAR INJECTION OF LABELLED RNA PRECURSORS

After injection of the tritiated RNA precursors [³H]guanosine, [³H]uridine or [³H]orotic acid into the eye of goldfish, labelled TCA-soluble material and RNA appeared to be axonally transported to the contralateral optic tectum. From the time courses of arrival in the tectum,‘average’rates of transport of 6 mm/day for the soluble material and 1·7 mm/day for the RNA were calculated. If the optic nerve was cut after the transported material had arrived in the tectum, about 60 per cent of the TCA-soluble material disappeared by 7 days after the cut, but almost none of the RNA. After a further 8- to 13-day period, the TCA-soluble material had declined by a further 50 per cent from the 7-day value, but the RNA by only 20 per cent. Thus, relatively little RNA was lost when the optic axons degenerated, an observation which suggested that the RNA might be extra-axonal. However, if the optic nerve was crushed before the arrival of the transported material, RNA did not appear in the tectum until the regenerating optic nerve fibres arrived. Therefore, the presence of RNA must be dependent on intact nerve fibres. Moreover, in the earliest stages of regeneration the proportion of transported RNA to TCA-soluble material was considerably higher than normal, suggesting that the regenerating fibres arrived in the tectum already carrying RNA. This implies that the RNA itself was transported in the optic fibres.     

Lipids in Cruciferae

Diploid and tetraploid types of white mustard (Sinapis alba) and turnip rape (Brassica campestris), annual and biennial forms, were grown in Turkey and in Sweden. Seed weight and oil content were measured and fatty acid composition determined by gas chromatography. Polyploidization of all cultivars effected an increase in seed size but did not markedly change the oil percentage or fatty acid composition. Growing conditions in Turkey caused an over-all decrease in seed size although the oil content was reduced only in the spring Turkish plantings. Fatty acid composition was influenced by the environment to a small but probably significant extent in all cultivars. For Swedish and Turkish winter turnip rape the mean content of oleic acid was 13.7 per cent and 15.8 per cent, of linolenic acid 9.5 per cent and 8.6 per cent, of erucic acid 45.7 per cent and 42.4 per cent, respectively; no consistent differences were observed for linoieic acid. Similar differences were observed for summer turnip rape and white mustard. Thus the warmer and drier Turkish climate favoured a higher oleic acid, lower linolenic and erucic acid content, i.e. a reduction in the per centage of the major end products of fatty acid biosynthesis.     

Phage formation in Staphylococcus muscae cultures. XI. The synthesis of ribonucleic acid,desoxyribonucleic acid,and protein in uninfected bacteria

1. The synthesis of ribonucleic acid, desoxyribomicleic acid, and protein in S. muscae has been studied: (a) during the lag phase, (b) during the early log phase, and (c) while the cells are forming an adaptive enzyme for lactose utilization. 2. During the lag phase there may be a 60 per cent increase in ribonucleic acid and protein and a 50 per cent increase in dry weight without a change in cell count, as determined microscopically, or an increase in turbidity. 3. During this period, the increase in protein closely parallels the increase in ribonucleic acid, in contrast to desoxyribonucleic acid, which begins to be synthesized about 45 minutes after the protein and ribonucleic acid have begun to increase. 4. The RNA N/protein N ratio is proportional to the growth rate of all S. muscae strains studied. 5. While the RNA content per cell during the early log phase depends upon the growth rate, the DNA content per cell is fairly constant irrespective of the growth rate of the cell. 6. Resting cells of S. muscae have approximately the same RNA content per cell irrespective of their prospective growth rate. 7. While the cells are adapting to lactose, during which time there is little or no cellular division, there is never an increase of protein without a simultaneous increase in ribonucleic acid, the RNA N/protein N ratio during these intervals being approximately 0.15. 8. Lactose-adapting cells show a loss of ribonucleic acid. The purines-pyrimidines of the ribonucleic acid can be recovered in the cold 5 per cent trichloroacetic acid fraction, but the ribose component is completely lost from the system. 9. The significance of these results is discussed in relation to the importance of ribonucleic acid for protein synthesis.     

CHANGES IN THE BASE COMPOSITION OF NUCLEAR RIBONUCLEIC ACID OF NEURONS DURING A SHORT PERIOD OF ENHANCED PROTEIN PRODUCTION

Nuclei from isolated nerve cells were sampled by microdissection. The content and composition of the nuclear RNA was studied and compared with that of the cytoplasmic RNA of Deiters' nerve cells of rabbits. Analyses were made of control nerve cells and of cells in which an enhanced RNA and protein production had been induced by chemical means, tricyano-amino-propene, for 60 minutes. The nuclear RNA content of the control nerve cells was 56 µµg, i.e. 3 per cent of the total RNA content of the nerve cell. The base ratios were: adenine 21.3, guanine 26.6, cytosine 30.8, uracil 21.3. Purine-pyrimidine analyses showed that the nuclear RNA differed significantly from the cytoplasmic RNA in having higher adenine and uracil values. The guanine and cytosine values were high, however, and the ratio G/C was 0.86 as compared with 1.16 for the cytoplasmic RNA. The composition of the nuclear RNA was interpreted as reflecting the extraordinarily strong development of the nucleolus in these neurons. During the 60 minutes of enhanced neuronal RNA production (+25 per cent) the guanine value increased and the uracil value decreased significantly in the nuclear RNA. In the cytoplasmic RNA the guanine value also increased although not so much as the nuclear guanine. The cytoplasmic cytosine value decreased. The result indicated that the production of the characteristic cytoplasmic RNA had been influenced by the change in the nuclear RNA