Wednesday, December 5, 2007

Classification of Enzymes

For many years it was the custom to classify enzymes solely by the reactions which they catalyzed. As already noted there is within living systems an amazing variety of chemical reactions and these many diverse reactions are specifically catalyzed by individual enzymes. However, as the chemical composition of enzymes becomes better understood, there is an increasing tendency to classify enzymes not merely in terms of what they do but also in terms of their true chemical composition.

In the first place it is possible to characterize a large group of enzymes as hydrolyzing enzymes or hydrolases. The hydrolases are concerned primarily with the breaking down and the building up of proteins, carbohydrates and lipids. It is a characteristic of these essential organic materials that their large molecules are built up by a progressive subtraction of water molecules. When amino acids unite, the acid group of one amino acid combines with the basic amino group of a second, and as in typical acid-base reactions, water is split off. In this way protein molecules are built up, and if they are to be broken down to amino acids again, water molecules must be added. Similarly, when an alcohol unites with an organic acid to form an ester, water is split off; and in the reverse process by the addition of water an ester may be converted into an acid and an alcohol. Thus by the addition of water, a fat may be hydrolyzed to fatty acid and the trivalent alcohol glycerol. When two molecules of a simple sugar such as dextrose unite to form a disaccharide, then also water is split off, and with the addition of water the double sugar may be broken into simple sugars again. Similarly starch is formed from sugar by a series of reactions in which water is removed. All of these reactions in which water is removed or added are catalyzed by the hydrolases. Proteolytic enzymes break down proteins to polypeptids and amino acids (or build up proteins from such compounds), lipases split fats into fatty acids and glycerol (or synthesize the fats from the fatty acids and glycerol), amylase acts on starch, sucrase (often called invertase) acts on sucrose, and there are other specific enzymes which act on the various isomers of sucrose. The hydrolases also include the lecithinases which split off fatty acids from the rest of the lecithin molecule, and the phosphatases. The latter split off phosphate from organic combination. Thus one phosphatase breaks glycerol phosphate into glycerol and phosphate; another group of phosphatase enzymes, the nucleotidases, break nucleotides into nucleosides and phosphoric acid. Most hydrolases are concerned primarily with the breakdown of large molecules as it occurs in digestion and with the synthesis of the breakdown products to form large molecules again in the cells. Such reactions are of primary importance in nutrition, but they yield little energy.

In addition to the hydrolases concerned primarily with nutrition, there are some hydrolases which play a part in other vital phenomena. Thus the enzyme cholinesterase, which hydrolyzes acetylcholine into choline and acetic acid, is believed to have great importance for nervous function. Another hydrolase enzyme which has excited great interest is hyaluronidase. This breaks down hyaluronic acid, a polysaccharide that tends to hold cells together. The enzyme is found in various bacteria; also in testis extracts. When skin is treated with hyaluronidase, the cells separate sufficiently to permit the spreading between them of substances like India ink. Heparin, like hyaluronic acid, is a mucopolysaccharide. There is apparently a heparinase which breaks up heparin or at least destroys its activity. Mention has already been made of the enzymes which specifically attack ribonucleic acid and desoxyribonucleic acid. These likewise are hydrolases.

The enzymes which cause coagulation or clotting, thrombin and rennin, may also be hydrolases. Thrombin causes clotting of vertebrate blood and rennin clots milk. They may both produce this effect as a result of proteolytic action.

Similar to the hydrolases are the phosphorylases. These enzymes, which should not be confused with the phosphatases, add phosphoric acid instead of water. They are very important in carbohydrate metabolism and will be discussed later in this connection.

Enzymes which act on inorganic pyrophosphates and hydrolyze them to orthophosphates are called pyrophosphatases. (This is a reaction which occurs even without the aid of enzymes.) Pyrophosphatases are found in many different types of living material.

In addition to the hydrolases there are various other categories of enzymes. There are enzymes which transfer a radical or group from one substance, the donor, to another, the acceptor. These have been called transferases, and they include the transaminases, which transfer amino groups; and the phosphokinases, which transfer phosphate groups.

An interesting enzyme found especially in the red blood cells of higher animals, is carbonic anhydrase. This catalyzes the reaction in which carbonic acid is split into carbon dioxide and water. Actually, the reaction is speeded in both directions, so that carbonic acid can be formed more rapidly as well as broken down. The reaction needs to be speeded in order to hasten the gain or loss of carbon dioxide from the blood.

Definition of Enzymes

The living machine is essentially a chemical engine, dependent for its growth, maintenance and energy on chemical reactions. These reactions are controlled by catalysts. One of the most striking achievements of modern biology and biological chemistry is the isolation of more and more of these catalysts so important for the vital process. Once isolated, it is possible to study their behavior in test tubes or other suitable containers. In other words, one can detach from the living material certain non-living substances capable of causing or promoting the complex chemical transformations which constitute a major part of the mystery of protoplasm.

Originally, enzymes were defined as "catalytic substances produced by living cells." With increase in knowledge and understanding, it has become necessary to restrict this definition. The original concept was that enzymes were not only produced by living things but were peculiar to them. There are some rather simple substances in protoplasm which can act catalytically; so for example, ascorbic acid, glutathione or as a matter of fact, even the hydrogen ion. The term enzyme was never meant to apply to these. Hence in modern usage an enzyme is defined as a "catalyst of biological origin, possessing a high molecular weight." In the light of what we now know, it is generally believed that all enzymes are proteins.

In as much as enzymes are catalysts, it is important to remember that they hasten rather than initiate reactions, and that when they act on reversible reactions they accelerate the progress of the reaction in either direction. Thus the same enzyme may synthesize as well as break down a complex chemical compound, and the very enzyme which digests proteins or fats or carbohydrates also serves, under the proper conditions, to build up these compounds from their breakdown products.

Digestion--Enzymes

Whenever an organism consumes food in the solid state, this must be brought into solution before it can be utilized. It is necessary, therefore, that such solid food be digested. In some organisms, as pointed out in the last chapter, digestion may occur outside of the body, and this may constitute an important adaptation for those animals which are in the habit of eating animals larger than themselves. Ordinarily, however, food is taken into the body and digested there. Digestion may occur in cavities of special organs such as the stomach or intestine, or it may occur within the protoplasm of cells. The latter type of digestion obviously takes place in protozoa. In organisms such as paramecium or ameba the ingested food is enclosed in a food vacuole, which serves the same purpose as the stomach or intestine of a complicated metazoan. Within the food vacuole the solid food particle is brought into solution. It must not be thought, however, that intracellular digestion is confined to protozoa. In sponges, coelenterates, and flatworms, much of the solid food taken into the body is ingested by ameboid cells lining the walls of the digestive tract, and digestion takes place within the protoplasm of these cells rather than in the lumen of the digestive tube or cavity. Moreover, in some animals rather higher in the evolutionary scale, there is also a certain amount of intracellular digestion. In spiders and in arachnids generally, the digestion of protein, begun either outside the animal or in the digestive tract, is finally completed within the cells lining the tract. In clams and other lamellibranchs, the digestion of protein and fat has been thought to occur exclusively within cells. Both in lamellibranchs and echinoderms, amebocyte cells play an important role. These phagocytic cells even enter into the lumen of the stomach or intestine, ingest particles of food there and then carry these food particles back into the tissues and digest them there. Such intracellular digestion, however, is a primitive character and it does not occur to any extent in higher animals such as the insects and vertebrates.

In the conversion of solid food to a state of solution, enzymes play the leading role. It will be necessary, therefore, to consider the subject of enzymes and the nature of enzyme action. It should be strongly emphasized, however, that enzymes are not concerned only with digestion, but that they are essential factors in all of the chemical activities of the organism. Our discussion at this point will be somewhat parenthetical. It will also be brief; indeed, it would scarcely be possible to present an up to date summary of enzyme chemistry in the space of a single chapter. The subject has grown so rapidly that it has become a science by itself (enzymology), and the modern books on enzymes are heavy with information.

Metazoa

Most many-celled animals take solid food into some cavity within their bodies, in which it is dissolved or digested. This is not always the case, however. There are quite a few organisms which are able to digest their food outside of their bodies. Thus a starfish or sea urchin may pour digestive juices out over some fish or other large animal which it has captured. It is sometimes possible for an animal to eat other animals larger than itself. When a spider captures a fly, it pours digestive juices into the body of its prey and then sucks out the dissolved interior. Many parasitic animals require no digestive apparatus. In the case of animals such as tapeworms, there is no need for the ingestion of solid food, and the dissolved food substances in the alimentary canal of the host pass directly through the body wall of the animal. Other worms derive food in the same manner from the blood of their hosts.

Some authors have tried to classify animals on the basis of their methods of ingestion. Thus there have been distinctions between whirlers (ciliary feeders), snarers, scrapers and suckers. Small particles are often pulled into the body by ciliary action. This occurs not only in the protozoa, but also in practically every other phylum, except the arthropoda. Ciliary mechanisms for feeding are most highly developed in the mollusca. Tentacles and setae are also used in obtaining small particles. The animals that feed on large particles or masses have mechanisms for swallowing inactive food, for seizing prey, or for scraping and boring.

The usual form of ingestion in metazoa involves some sort of passage into the mouth. Jaws and teeth of various types may aid in the capture and in the subdivision of the food. The earthworm has a pharynx which is pulled on by muscles attached to its outer wall so that it can behave as a suction pump. Marine (polychaete) worms also may have sucking pumps. Thus in Autolytus the proventriculus, by its pulsations (120 per minute), produces a strong inward current of water and food. Sucking pumps also occur in insects of various orders, and these pumps may have a rather complicated structure. Even in the case of those animals which ingest food through the mouth there is sometimes a possibility that food may be taken in through the body wall.

Feeding habits of protozoa

Feeding habits of protozoa vary widely. Some contain chlorophyll and behave essentially like plants. Thus the various species of Euglena are able to manufacture starch from simple inorganic materials. These forms and also ciliates without mouths, such as the Opalinidae, are able to absorb food materials through their surfaces. In all of the sporozoa, moreover, food enters the organism through the body wall. These forms are parasites and live on the organic materials provided by their hosts.

Most ciliates and flagellates have definite mouths, but it does not follow that all of the food taken up by these organisms passes through the mouth opening. In flagellates, especially, it seems probable that much of the food enters the cell by diffusion through the body wall. Ciliates like paramecium sweep food particles into their gullet by the beat of their cilia. Although various useless materials may enter the gullet and pass into the cell, there is apparently some power of discrimination.

The ingestion of food by ameba is essentially the same process as that which occurs when leukocytes or other ameboid cells take up foreign particles. Such phagocytosis occurs in practically all metazoa, and is of great importance both in the normal life of animals and in the resistance of animals to disease. Phagocytosis plays an essential role in metamorphosis and, both in insects and in amphibia, larval organs are destroyed bit by bit by the ameboid cells which ingest them. Because of its importance as a protection against disease, phagocytosis has been very widely studied by bacteriologists, pathologists, and students of medical sciences generally. Just as amebae select their food, so do leukocytes exercise a selection of the particles they ingest. The rate of ingestion of bacteria by leukocytes has often been studied under a variety of conditions. Bacteriologists have tried to obtain quantitative data by determining either the number of bacilli taken up per leukocyte or the percentage of leukocytes participating. Important factors (not always properly considered) are the absolute and relative concentrations of bacteria and leukocytes. Phagocytosis increases with increase in the number of bacteria, but the rate of ingestion does not keep pace with the increasing possibilities for collision between bacteria and leukocytes. Various authors have studied the effect of temperature on phagocytosis; curves are obtained like those found for ingestion of food by ameba. Any explanation of the mechanism of phagocytosis could at the same time serve as an explanation of the ingestion of food by ameba. At the present time, however, none of the theories of phagocytosis seems very satisfactory.

In higher animals, phagocytosis is not restricted to leukocytes. Scattered through various tissues and organs of the animal are numerous cells capable of ingesting solid objects. Of these, the macrophages move somewhat more freely than the histiocytes, and there are also morphological differences between the two types of cells. Together, they constitute the reticuloendothelial system.

Vitamins of the B Complex

B 13 has been described as a growth factor for rats and for pigs. There is also a description of a vitamin B 14, extracted from urine.

Over the years, there has been a large and controversial literature about what has been called vitamin P. Substances with vitamin P activity are supposed to maintain proper permeability of the blood capillaries of higher animals and to prevent excessive loss of fluid from them. (The letter P is due to this permeability action.) There is now general agreement that substances with P activity are not to be considered as vitamins. At least this is the judicial opinion of the American Society of Biological Chemists.

Para-aminobenzoic acid (often called paba). This relatively simple compound is an important growth factor for many bacteria. As previously noted, it forms part of the folic acid molecule. Para-aminobenzoic acid and sulfanilamide are very close chemically, as is apparent from their structural formulae:

Thus sulfanilamide is an antagonistic analogue of para-aminobenzoic acid, and its antibacterial action is due to this similarity in structure. Choline. This substance, the basic constituent of lecithin, is believed by many to be a vitamin. Rats deprived of choline develop fatty livers and other types of hepatic injury. They may also show degenerative changes in the kidney. Choline deficiency is also a factor in the development of a leg deficiency (perosis) in chicks and turkeys.

It should be remembered that choline is a constituent not only of lecithin, but also of the important humoral substance, acetylcholine. Inositol. Inositol is a derivative of benzene. It can be prepared from hexahydroxy-benzene by reduction, in the course of which 6 hydrogen atoms are added. Thus inositol is hexahydroxy-hexahydro-benzene. It is an optically active substance found in yeast, muscle, and in various other types of animal and plant tissue.

Inositol is important for the growth of yeasts, and is indispensable for some types of yeast. It also promotes the growth of some fungi. In the presence of inositol, chicks grow more rapidly than when it is absent. However, rats may be bred for three generations without inositol, and György was led to conclude that "the facts presented do not, at the present time, warrant the identification of inositol as a primary and really essential vitamin; they rather favor the assumption that it is a supporting and at least not always specific vitamin." a-Lipoic Acid. This substance, also called protogen, is probably to be included among the vitamins of the B group, for it is water soluble. It is important for the growth of some bacteria and for the protozoan Tetrahymena, and it is involved in the oxidation of the important keto acid, pyruvic acid (for this reason it was formerly called pyruvate oxidase factor). Many types of living material contain a-lipoic acid. It is now known to be a derivative of octanoic (caprylic) acid and it contains sulfur in the form of an S-S group.

Throughout the course of vitamin study, there have indeed been many extravagant claims and many false leads. The vast literature is often contradictory and confused. But in spite of this, in the course of time the truth has gradually emerged and the main outlines of the picture are now clear. All sorts of organisms, from bacteria to man, require certain specific types of organic molecules. These can be synthesized by green plants and by many lower organisms. Higher forms have frequently lost the power for such synthesis, and unless, like the ruminants, they provide themselves with huge storehouses for bacteria, they must depend for their existence on obtaining the vitamins with their food.

From the knowledge of the nature of the vitamins, we can begin to develop a chemical anatomy of living systems. The vitamins represent indispensable organic molecules and apparently the same molecules are almost universally needed. Moreover, we are beginning to discover why the vitamins are needed and what work they do in the living cell.

Much of our information is still fragmentary. The vitamin requirements for the lower vertebrates and for the hosts of invertebrate animals have scarcely been investigated at all. Moreover, in some instances we are completely at a loss to account for the fundamental necessity for a given vitamin. Such superficial disturbances as a reddening of the eye or a scaliness of the skin give no direct indication of the part a vitamin may play in protoplasmic activity.

Vitamin B 12

Year by year, the number of B vitamins increases. B 10 and B 11 are required by chicks, the former for proper feathering, the latter for growth. From liver it has been possible to isolate a pink crystalline substance which can be used to cure pernicious anemia. Folic acid is useful in this disease, for it has a hematopoietic effect (that is to say, it favors increased production of blood cells), but it does not correct the nervous symptoms of the disease. These depend on degenerative changes in the nervous system. Vitamin B 12 in tiny amounts has been thought to correct diseases of both blood and nervous systems, but apparently if the nervous system is badly affected, it can not be cured. The vitamin has been crystallized. It contains 4-4.5 per cent cobalt and also phosphorus; the molecular weight is approximately 1500. (Estimates vary from 1300 to 1600.) In liver extracts there are really two active substances with slightly different absorption spectra. One of these is the definitive B 12, the other is B 12b. (B 12a is another similar substance derived from B 12 by hydrogenation.)

Vitamin B 12 is apparently identical with what has been called "animal protein factor." This was originally obtained from such diverse sources as milk, muscle, liver and cow manure. Although called animal protein factor, it can likewise be obtained from plant sources. Both animal protein factor and vitamin B 12 promote growth in some bacteria, in chickens, rats and mice. The effect on growth may be due to the fact that the vitamin seems to increase the utilization of amino acids. At any rate, this has been reported for chicks.

Folic acid

This vitamin occurs in green leaves; hence the name, which is taken from the Latin word for leaf. Folic acid also occurs in mushrooms and yeast; and in animal tissues, such as liver and kidney. The vitamin has been called by many different names, and it exists in various forms. These have different potencies for different types of organisms.

Chemically, folic acid consists of pteroic acid combined with the common amino acid, glutamic acid. Pteroic acid itself is a combination of para-aminobenzoic acid and a pteridine base. Pteridine bases are pyrimidine derivatives related to the pyrimidine bases thymine and uracil found in the nucleic acids. The formula of folic acid is given below:

The compound shown above is pteroylglutamic acid, sometimes called PGA. Pteroic acid may also combine with 3 glutamic acid groups to form pteroyltriglutarnic acid (P3GA) or with 7 molecules of glutamic acid to form pteroylheptaglutamic acid (P7GA). All of these compounds are found naturally and all have vitamin activity for certain types of living materials.

Folic acid is required by some bacteria, by certain ciliate protozoal, by some insects and by various birds and mammals. The bacteria in the rat intestine normally manufacture a sufficient quantity of the vitamin, and treatment with antibiotics such as sulfa drugs is necessary to cause folic acid deficiency. Folic acid is commonly an essential growth factor, not only for bacteria but for higher organisms as well. Lack of the vitamin in mammals results in various ailments, but especially in a type of anemia characterized by a small number of red blood cells of large size. This type of anemia, called macrocytic anemia, occurs in the tropical disease sprue, and sprue can be cured either by liver extract (which contains folic acid) or by folic acid itself. Folic acid is apparently important both for growth and cell division. When chick embryos are in contact with antagonistic analogues such as 4amino folic acid, their growth is greatly retarded. Similar results have been obtained in numerous other studies with folic acid antagonists. Thus 4-amino folic acid has a retarding effect on tumor growth.

A substance related to folic acid is the so-called "citrovorum factor," 45 which is apparently identical with "folinic acid." This factor favors growth of the bacterium, Leuconostoc citrovorum. The citrovorum factor may be obtained from liver extracts; also it is excreted into the urine of rats or men fed folic acid.

Biotin

As long ago as 1898 it was shown that if dogs were fed raw egg white, diarrhea resulted. Following this it was shown that other mammals also suffered from egg white feeding, and that in addition to the diarrhea, skin ailments and nervous symptoms appeared. The phenomenon became known as egg white injury. Cooking the egg white prevented the injury. György proposed the term vitamin H for a factor which prevented egg white injury. Later, it was shown that this factor is identical with biotin, a vitamin known to be important for yeasts and microorganisms. The raw egg white unites with the biotin and prevents its utilization by mammals or birds.

Chemically, biotin is rather complicated. It has in its molecule a urea ring containing one sulfur atom; the ring is combined with a valeric acid group. The presence of sulfur in the molecule is not important for the vitamin activity, for if the sulfur is replaced by oxygen, the vitamin activity is retained. The formula of biotin is given below:

Biotin acts in extremely low concentration, and it is thus one of the most potent of the vitamins. Many organisms do not grow unless biotin is available for them. Usually in higher organisms it is synthesized in sufficient quantity, but biotin deficiency can occur naturally in birds and in insects. Lack of sufficient biotin has the same effect as egg white injury--diarrhea, disturbances in the skin and nervous system, in rats loss of hair about the eyes. It can also prevent the birth of young in rats. The protein in egg white which binds biotin is called avidin; it is a basic, carbohydrate-containing protein and it occurs not only in the eggs of birds but also in frog eggs.

There is now strong evidence that biotin is important in a process which is known as carbon dioxide fixation. Many bacteria, and higher organisms also, are able to utilize carbon dioxide and form organic compounds from it. This process is interfered with if biotin is lacking.

Pantothenic acid

Pantothenic acid is found in many diverse types of living materials. Chemically, it consists of ß-alanine in a peptid linkage with a dihydroxy acid. (ß-alanine differs from ordinary alanine in being ß-aminopropionic acid instead of α-aminopropionic acid.) The formula of pantothenic acid is given below:

The acid is unstable and for this reason is sold as a calcium salt. This salt is a white powder, readily soluble in water but not in alcohol.

Many organisms from bacteria and protozoa to higher mammals require pantothenic acid. In its absence these organisms do not grow properly and they may show various ill effects. Symptoms of pantothenic acid deficiency include dermatitis (inflammation of the skin) in rats, deficient feathering in chicks, graying of hair in rats, degeneration of the nervous system in pigs, hemorrhages in the adrenal gland and general disturbance of function of this gland, etc.

The vitamin constitutes part of an enzyme system. Combined with various substances including phosphate, adenylic acid, and a sulfhydryl group, it is coenzyme A, and this in combination with a protein is an enzyme capable of acetylating various organic substances such as choline and sulfanilamide. The acetylation of choline may be very important physiologically. How the acetylation, or rather the lack of it, can produce all the pathological conditions noted above is not clear, although in the case of the nervous system, any alteration of acetylcholine metabolism would certainly be harmful.

Vitamin B6: pyridoxine, pyridoxal, pyridoxamine

The term vitamin B6 is now properly used, according to a decision of the American Society of Biological Chemists, to designate three naturally occurring substances: pyridoxine, pyridoxal and pyridoxamine. The formulae of these substances are given below:

Pyridoxine is 2-methyl-3-hydroxy-4,5-dihydroxymethyl pyridine. Pyridoxal is the aldehyde derivative of pyridoxine and has a formyl group instead of a hydroxymethyl group at the 4 position. Pyridoxamine is an amino derivative and has an amino methyl instead of a hydroxymethyl group at the 4 position. The three forms of the vitamin are almost equally effective for higher animals, but some bacteria
have specific requirements for one form or the other. Vitamin B 6 is needed by many bacteria, by some protozoa, and by most of the insects whose vitamin requirements have been investigated.

When rats are deprived of vitamin B 6, they develop a painful inflammation of the skin. This is especially evident in the skin of the extremities--the paws, snout, and ears. The ailment is sometimes called rat acrodynia because of a supposed similarity to a human ailment in which the nerves and skin of the hands and feet are affected. The disease is not wholly specific for vitamin B 6 deficiency; it can be produced also in other ways, for example by a lack of unsaturated fatty acids. In various higher animals, convulsions may follow extreme vitamin B 6 deficiency. Sometimes lack of the vitamin causes muscular weakness; it can also cause anemia.

Vitamin B 6 deficiency can be induced by the addition of an antagonistic analogue, desoxypyridoxine. Thus if this compound is injected into hens' eggs, the embryos do not proceed very far in their development.

Vitamin B 6 is important for the proper metabolism of the amino acids. It forms part of an enzyme system which promotes the synthesis of amino acids by the addition of carboxyl groups. Acting as part of what is presumably the same enzyme system, the vitamin also favors the removal of carboxyl groups from amino acids. Another type of enzyme for which vitamin B 6 is an important constituent is transaminase. This enzyme catalyzes the transfer of an amino group from one amino acid to another; an important type of reaction in protein metabolism. Thus there is some important information as to how the vitamin functions with respect to the chemical reactions occurring during the metabolism of proteins and amino acids, but how this is related to the disturbances which occur in skin, nerves, muscle and blood when the vitamin is lacking has yet to be explained. It has been claimed, on the basis of isolation studies of mitochondria, that these granules contain especially large amounts of vitamin B 6. However, it is the nucleoprotein-containing granules of the cytoplasm that are believed to be especially important for protein metabolism.

Nicotinic acid: Niacin

Nicotinic acid and pyridoxine are closely related substances. Both are derivatives of pyridine. This substance is a weak base, the molecule of which contains five CH-groups and a nitrogen atom arranged in a six-membered ring. As is customary with ring compounds, the various positions on the ring have numbers. In pyridine, nitrogen occupies the number 1 position. Nicotinic acid is 3-pyridine carboxylic acid. The structural formulas of the acid and its amide are given below:

The amide is more important than the acid itself, for some organisms require it and cannot manufacture it from the acid. Both acid and amide are white crystalline powders, without odor.

Nicotinic acid is one of the commonest growth factors needed by bacteria; in other words the ability to synthesize it is one most frequently lost. This may be due to its very wide distribution in soil and natural media. It is an essential requirement for those ciliate protozoa that have been studied (Colpoda and Tetrahymena) and also for most of the insects that have been investigated. Nicotinic acid is essential for the dog, the pig, and for man. However, if enough tryptophan is added to the diet, neither dog nor pig requites it. As noted previously, tryptophan is a precursor of nicotinic acid, and the tissues can synthesize the vitamin from tryptophan.

From a lack of nicotinic acid, dogs become afflicted with what is known as blacktongue disease. In man, nicotinic acid deficiency is the primary cause of pellagra. This serious disease, common enough in the United States, involves the lining of the digestive tract, the skin, and the nervous system.

Nicotinic acid amide enters into the composition of two substances of great importance for the oxidative metabolism of the cell. These are coenzyme I and coenzyme II. Presumably nicotinic acid deficiency is associated with a lack in the tissues of coenzymes I and II. But what connection there is between such a lack and the manifold disturbances present in a disease like pellagra is not at all clear.

Riboflavin Vitamin B2 Vitamin G

Riboflavin is an orange crystalline substance, sparingly soluble in water, and markedly sensitive to light. The riboflavin molecule is complex. The structural formula follows:

The three rings represent an isoalloxazine constituent and this is combined with ribose, the pentose sugar also present in ribonucleic acids.

Although riboflavin is apparently a constituent of all or almost all types of protoplasm, many lower organisms do not require it in their food. Yeasts and molds have no need for it. Bacteria, in general, can get along without it. However, riboflavin serves as a growth factor for the tetanus bacillus, for lactic acid and propionic acid bacteria, and it is a requirement for Erysipelothrix rhusiopathiae and Listerella monocytogenes. The protozoan ciliates that have been investigated require riboflavin. So, too, do almost all the insects that have been studied. Riboflavin is essential for the growth and normal health of the rat, mouse, dog, pig, man, chicken and turkey. In riboflavin deficiency, various symptoms of ill health may appear. These include redness and roughness of the cornea of the eye, soreness at the corners of the lips, redness and soreness of the tongue, etc. At one time, it was thought that "ariboflavinosis," the disease caused by riboflavin deficiency, was extremely common. But the symptoms mentioned above are not always due to lack of riboflavin.

The presence of riboflavin in cells may be detected by fluorescence microscopy. Riboflavin is important for cell respiration. It and other alloxazine compounds combine with phosphoric acid and protein to form enzyme systems known as flavoproteins.

Thiamine: Vitamin B1

The formula for thiamine is C 12 H 18 N 4 OS. It is prepared synthetically as the chloride hydrochloride. This is a white crystalline compound readily soluble in water. Thiamine is a combination of a pyrimidine ring and a sulfur-containing thiazole ring. The structural formula of the chloride hydrochloride is as follows:

Apparently all sorts of protoplasm need thiamine. Some types of cells can synthesize it, others can not. Higher organisms generally require the intact thiamine molecule; on the other hand some lower organisms are able to synthesize thiamine provided they can obtain both parts of the molecule, pyrimidine and thiazole. Other organisms may require either the pyrimidine fraction or the thiazole fraction, but not both. Many bacteria require neither and can synthesize whatever thiamine they need.

Lack of thiamine in the diet produces beriberi, a paralytic disease of men rather common in Japan and in various other portions of the world. In birds, lack of thiamine produces polyneuritis, a disease similar to beriberi. In this disease the nerve fibers show degenerative changes.

Experimentally, it has been shown that a similar paralysis could be produced in chicks, cats, pigeons and rats. The effect is due to the fact that some species of fish contain in their tissues an enzyme, thiaminase, which destroys thiamine. Thiaminase is found not only in fish but also in clams. In addition to its effect on the nervous system, lack of thiamine may also cause retardation of growth. Thiamine is important for the utilization of carbohydrate. Combined with 2 phosphate groups as diphosphothiamine, it forms part of an enzyme system that acts on pyruvic acid, a keto acid of great importance in intermediary metabolism. When animals are deprived of thiamine, their tissues are unable to oxidize pyruvic acid. Thiamine is also important in the transmission of the nerve impulse. When isolated nerves are stimulated, they are believed to release thiamine to the surrounding solution.

Ascorbic acid: Vitamin C

The fat-soluble vitamins are still generally referred to by their alphabetical names. On the other hand, the water-soluble vitamins are now for the most part usually given chemical names which have no relation to the alphabet. This became necessary when what was originally the B vitamin became a whole assortment of heterogeneous substances, distinguished by somewhat variable and uncertain subscripts of the letter B. At present the water-soluble vitamins include the clearly defined ascorbic acid (vitamin C) plus the vitamins of the B complex. Because of the fact that vitamin C is one substance, clearly defined and understood, we shall consider it first rather than in its alphabetical order.

Ascorbic acid is now well understood chemically and it can indeed be synthesized. The structural formula of the L-form is:

The L-form is much more potent than the D-form. Ascorbic acid is really an acid hexose sugar. Its acidity is due to the dissociation of an enolic hydrogen rather than to opening of the lactone ring.

Ascorbic acid is a strongly reducing substance, and because of this, it is easy to demonstrate its presence in cells and tissues. A standard method involves the reduction of silver nitrate. Because of its reducing power, there is doubtless a relation of ascorbic acid to the oxidative processes in cells, but there is still no clear understanding as to the exact function or functions of the vitamin in metabolism. In animals suffering from scurvy there is lack of intercellular material; presumably the cells enclosing the blood vessels are not held together as firmly as they should be and this is doubtless the cause of hemorrhage. Mucopolysaccharides are known to be important constituents of the cementing substances which hold cells together. In view of the fact that mucopolysaccharides are polymers of acid sugars, it is possible that ascorbic acid is involved in the synthesis of such substances. There is some indirect evidence for this. Normally in wound healing mucopolysaccharides are detectable in the healing tissues and may have considerable importance for the process. However, when the store of ascorbic acid is depleted, these substances do not appear.

Vitamins K: fat-soluble vitamins

Fourth and last of the commonly recognized fat-soluble vitamins are the vitamins K, so named because they are important for blood coagulation (coagulation is spelled with a K in German and in the Scandinavian languages). The K vitamins are close to the E vitamins chemically. Tocopherols are very similar to the naphthoquinones chemically; the K vitamins are actually naphthoquinones.

The basic structure of a naphthoquinone is shown below:

This is a 1,4 naphthoquinone. The most active form of vitamin K, a form which can be prepared synthetically, is 2-methyl-1,4-naphthoquinone. This is sometimes called menadione. It does not occur naturally nearly as abundantly as vitamin K1, which resembles menadione except that instead of having a methyl group in the 3 position, it has a phytol group. (Phytol is a 20 carbon mono-unsaturated primary alcohol which can be obtained from the saponification of chlorophyll.) Vitamin K 2 also resembles menadione except that in the 3 position it has a difarnesyl group. (Farnesol is similar to phytol; it is a 15 carbon primary alcohol with 3 unsaturated carbon atoms.) Several other naturally occurring vitamin K compounds have also been described. They are all either yellow oils or yellow crystalline solids, practically insoluble in water.

Vitamin K is manufactured in one form or another by bacteria, which do not ordinarily require it. The bacteria in the intestines of mammals produce enough so that normally the mammals have no need of it from outside sources. However, when absorption of fatty compounds is interfered with, as in intestinal disease or in liver disease involving interruption of bile flow, the vitamin becomes essential for man and mammals. Many birds require the vitamin, for their bacteria can not supply them with sufficient quantity.

Vitamin K is necessary for the clotting of the blood of higher animals. The reason for this is not clearly understood. To some extent opinions differ, but most authorities believe that this action of vitamin K is an indirect effect caused by the entrance into the blood of greater amounts of a substance prothrombin, the precursor of thrombin. Thrombin, which is regarded as an enzyme, is of primary importance in blood clotting.

Certainly the function of vitamin K can not be restricted to its effect on the clotting of blood. Obviously, the bacteria and the plants which produce it must have some need for it themselves. Experiments with isolated cells show that vitamin K is a very powerful protoplasmic clotting agent. This is not surprising in view of the fact that the clotting of protoplasm is in so many respects similar to the clotting of blood. Although menadione is but sparingly soluble in water, solutions of it cause a coagulation of the protoplasm of marine eggs. Menadione also produces clotting in muscle protoplasm.

A fat-soluble vitamin: vitamin E

A third fat-soluble vitamin or group of vitamins is vitamin E, or perhaps one should say the vitamins E. There are very many substances with vitamin E activity, but the term vitamin E is restricted to the tocopherols. (The word tocopherol is derived from Greek words signifying child birth.) There are 4 tocopherols now known: alpha, beta, gamma and delta tocopherol. The formula for alpha tocopherol is:

Beta and gamma tocopherol have 1 less methyl group than alpha tocopherol and delta tocopherol has 2 less methyl groups.

Vitamin E is necessary for the reproduction of the rat and other mammals and birds. If the male rat is fed a diet lacking in vitamin E, the germ cells in the testis degenerate. In the absence of vitamin E, the female rat cannot give birth to its young; these degenerate and are resorbed. In some cases, muscular atrophy follows lack of vitamin E in the diet.

The mechanism whereby vitamin E functions in the body is still unknown." In the first place it may act as part of an enzyme system, and to this action its effect in preventing degeneration of muscle may be due. Secondly, the tocopherols exert another, less specific function in that they tend to prevent excessive oxidation. Thus, for example, they tend to preserve vitamin A and they may also prevent the necessary unsaturated fats from being oxidized.

Sterols - Vitamin D

A number of sterols have vitamin D activity, that is to say they prevent the occurrence of rickets in higher animals. The chemistry of the sterols is very complicated, and books on physiological chemistry may be consulted for structural formulae, etc. When the common sterol of fungi, ergosterol, is irradiated with ultraviolet rays, another sterol, calciferol, is produced. This is commonly referred to as vitamin D 2. (There is no D 1, for the original use of the term was a misnomer.) D 3 is 7-dehydrocholesterol. Numerous other antirachitic (rickets-preventing) substances have also been described. Whether or not a given sterol or sterol derivative has antirachitic action can only be determined by actual test.

Bacteria do not seem to require sterols. Some species of protozoan flagellates do; also Entamoeba histolytica, the ameba that causes human dysentery. Insects, in so far as they have been studied, require sterols, but they seem to need cholesterol, a cholesterol derivative, or a plant sterol; calciferol cannot usually be substituted for these.

In mammals and birds, vitamin D causes an increase in the deposition of calcium and phosphorus in the bones. This is apparently due, in part at least, to an increased absorption through the intestine. Using radioactive isotopes, Greenberg found that vitamin D promotes the absorption of calcium and strontium from the digestive tract. Sterols may act in the same way in promoting the passage of calcium through the walls of the insect intestine. Moreover, they may be concerned with the entrance of calcium into cells. This aspect of the subject has never been investigated.

Vitamin A is a yellow viscous oil

Vitamin A is a yellow viscous oil. Chemically, it is related to carotene, the common yellow pigment of plants. The generally accepted formula follows:

When carotene is eaten, it is converted to vitamin A. Because of this, it is sometimes called a provitamin. Another provitamin of vitamin A is kitol, a dihydric alcohol found in whale liver oil. The conversion of carotene to vitamin A occurs in the intestines of higher animals. The vitamin is then stored in the liver. The livers of polar bears and Arctic foxes are so rich in vitamin A that they are toxic. The toxicity is an example of what is known as hypervitaminosis.

Vitamin A has been synthesized in several different laboratories. There are really two forms of the vitamin. One form (A1) constitutes most of the vitamin content of the livers of salt water fishes. In the livers of fresh water fishes vitamin A 2 occurs. The 2 substances give slightly different colors with antimony trichloride and can be distinguished spectroscopically.

In higher animals, vitamin A is important for growth. When rats are deprived of vitamin A, the cornea of the eye becomes horny. This condition is known as xerophthalmia. It can occur also in children. Not only the corneal epithelium, but other types of epithelia are also adversely affected in vitamin A deficiency; this may lead to lessened resistance to infection. Vitamin A has a relation to vision. In man, vitamin A deficiency causes night blindness, but the effect is not as readily produced as formerly supposed.

In so far as they have been investigated, invertebrates, with the apparent exception of the snail Helix, do not seem to require vitamin A. Aside from its relation to vision, little is known as to the reason why vitamin A is necessary. Its presence in cells can be recognized with a fluorescence microscope because of the fact that in ultraviolet light it gives a green fluorescence.

Vitamins, riboflavin, nicotinic acid, pantothenic acid form

Most of the work on vitamins has been stimulated by the practical needs of medicine and industry. The physician and the drug manufacturer are not ordinarily interested in the vitamin requirements of lower organisms, unless these organisms can conveniently be used in vitamin assay. Thus, as might be expected, there are vast gaps in our knowledge of the vitamin requirements of many large groups of animals. We know almost nothing of the vitamin needs of metazoan invertebrates other than insects. And we know very little about the vitamins necessary for fish, amphibia and reptiles. A wide field is open for such study, and many facts of interest may well be discovered. Some vitamins are required by practically all living organisms from bacteria to man; others are needed by only some forms and not others.

There has been much interest in the avitaminoses, that is to say the diseases caused by the lack of specific vitamins. Our knowledge in this field is almost entirely confined to higher animals. Thus we know that when the vitamins D are omitted from the diet, a child will develop rickets and so will a pig or a rabbit, a dog or a chicken. Accordingly, the vitamins D have been called the antirickets vitamins. But animals without bones such as insects or protozoa may also require vitamin D. From a knowledge of the reasons why this is so, we may be able to obtain additional information as to the way that vitamins function.

Much as is known about the vitamins, their chemistry, their occurrence in various foods, the diseases that follow their absence from the diet, there is still almost complete ignorance as to the exact nature of their action. A beginning has been made in the recognition of the fact that some vitamins such as riboflavin, nicotinic acid, and pantothenic acid form essential parts of some enzyme systems. We know also the end results of many types of vitamin deficiency. But to say that a vitamin is necessary for growth or that it prevents anemia tells us very little as to what it really does or why it is necessary. If we were to know how certain vitamins influence growth or prevent cell deterioration in the epithelial or nervous system, we would have basic information of great importance. Such information would be useful not only in the better understanding of the vitamins but also in the elucidation of various basic vital processes. The final interpretation of vitamin action must lie in an understanding of how the vitamins affect the cell protoplasm.

It is this aspect of the subject that will be stressed in the following discussion of individual vitamins. The general physiologist is not primarily interested in the relation of vitamins to human welfare, the foods a man should eat, or the benefits to be derived from adding this or that vitamin to the diet. Such information can be obtained in books on physiological chemistry, nutrition, or medicine. In discussing the individual vitamins, we shall not proceed in alphabetical order. This order has no other basis than the chronology of discovery. We shall consider first the fat-soluble and then the water-soluble vitamins. In general, especially in the case of the water-soluble vitamins, true chemical names have tended to supplant letters and subscripts. But in some instances, several chemical substances can and normally do supply a given vitamin need. In these instances the alphabetical name is sometimes more convenient.

Sunday, November 18, 2007

About intercourse

In any case, if you and your teen-ager do manage to talk frankly together, sooner or later you will come to the question of sexual intercourse.

Like Betty, your child will doubtless have managed to gather the general facts. So also has Craig.

"The man puts his penis inside the woman," he states concisely. "I know that. But just where?"

Like Craig and like Betty, your child also will be after details. These, as you know now, you'll do well to explain in your own simplest and most comfortable way. Don't be afraid to draw from your own experience. And when you don't know, don't be afraid to say so, with a promise to consult book or doctor to find out.

"Which hole?" . . . You know now that to either boy or girl it's important knowledge that a woman has a third opening with a special channel or tube leading up from it to the uterus where her babies will grow. (We've talked about this already.)

"How is it done?" . . . You know this too.

"How often?" . . . It depends . . .

"But" said Nell, "you can't have that many babies."

"It isn't done every time to have a baby," her mother assured her. "It makes two people who love each other dearly feel closer and more intimate, more at one with each other. It's done for love many times . . . How often depends on how they both feel."

"How do they feel?" . . . Here is the tough one, the one at which the boy or girl invariably seeks to arrive if he dares.

But after you've considered it, you'll see that this one isn't too difficult either!

You'll need first to recognize what the adolescent is really after.

Actually he knows how it feels. He has known a long time.

The answer that your teen-ager usually wants to this question is not information or fact, but sanction for the feelings he has had in touching himself. For he intuitively knows inside him that these feelings are like those he will have in intercourse. He is now after reassurance; to know that his feelings are normal, healthy and good.

When Betty came to this question, her mother answered, "Those feelings? You know them. They're like when you look at a romantic scene in the movies or like when you touch yourself. Only even better. Because you're together with someone you love."

In this simple way of putting it, Mother shows Betty once more that Betty's experience with similar feelings has been wholesome and sound.

Betty gazed thoughtfully out of the window. And then she said an astonishing thing.

"That helps a lot, Mother. I'd been wondering whether I could hold out till I got married. Lots of girls don't. I think they're curious. Now that I know what it feels like, I won't have to do so much research."

"My!" thought Betty's mother, "haven't both of us grown up tremendously since we've talked on and off about these things over the last couple of years?"

How to help these adolescents not to be driven to do so much research? How to help them feel secure and satisfied enough to maintain controls? This is probably the biggest of all questions in the minds of parents. It, too, is part of what must be included in the sex education of today's youth.

So let us turn to it next and consider it in the context of rising interest in each other that boys and girls feel at this age.

If easy does it comes hard to us

Ideally what our boys and girls want is someone who can talk with them comfortably about sex. Someone who is not embarrassed. Someone who becomes neither bothered nor excited over the subject. Someone who is easy within himself.

We have repeatedly read these specifications! They are the ideal ones no doubt. But we haven't been brought up ideally. And not many of us can reach this ideal.

However, there are some things we can do to help us feel at least somewhat more comfortable.

The first of these takes us back to the matter of language. Betty's mother used the funny old little-girl term. That made her feel more comfortable because it was more familiar. On the other hand, for some people the less familiar words are the more comfortable ones. In speaking of bodies, for instance, some feel easier using such terms as "vulva" to designate the whole external female genital area, "vagina" for the opening and the channel inside, "clitoris" for the "small, humpy place that has such good feelings," as one girl described it, and "penis" and "testicles" or "testes" for the male organs.

And so as the first step to help get you at ease, choose whatever terms are the most comfortable for you. (In spite of the advantages to your child of using familiar terms, your maximum comfort--which may be shaky anyway--comes first!)

The next thing is to be frank about your feelings concerning the terms your child uses. If you can accept them sincerely, so much the better, as we have said. But don't be insincere. Insincerity makes you less comfortable. Moreover, your distaste will be sensed and will become a deterrent to frankness between you.

It's better to stop your child if his language distresses you (even though you know again this isn't ideal). One mother said, for instance, "I know you and some of your friends use those words. But I don't personally. And I've never liked them. I get too embarrassed. So let's not, around me."

Supply your teen-ager with words if he fumbles. But don't make your condemnation of his words an over-all, indiscriminate business. Let him know you know that some people use such terms decently and that your feelings against them are personal with you.

This brings us to another point: If you feel embarrassed on any score in talking about sex, better say so!

Right straight out: "I feel uncomfortable talking about sex! I wish I didn't. But I do."

"I still feel a bit tense!" confessed Betty's mother, when Betty came with more wonderings several months later. "When it comes to the sixty-four dollar question that we've never really gone into . . ."

Betty giggled. "Yes, Mom, you've always blushed whenever we've come within miles of that one! We almost got to it a couple of times and I could feel you backing away."

"Or turning and running!"

Then Betty's mother managed something else that held value. You can try this one also: Be frank about the mistakes and embarrassments of the past instead of evading them.

"Like the time you spelled out that four-letter word and asked me what it meant!"

"Yes," nodded Betty. "You got red as a beet and said I shouldn't say anything so horrible ever again. So I made up my mind I'd never ask you anything again! My! I was mad . . ."

"Uh-huh," thought Betty's mother. She knew, as you no doubt also know by now, that it's good to: Let your child get his old hostilities out for all the times you have not attended adequately to his sex education. It may help clear the air and make him more friendly. And if he's more friendly you'll be more relaxed.

"After all, I felt hostile to my folks for holding out on me. Why shouldn't Betty feel angry at me for having held out on her?"

She hadn't forgotten, and you don't need to either, that it helps to: Try recalling how you felt when you were young. This can bring more understanding of how your child feels right now.

Out loud Betty's mother continued: "I was stupid to renege on that most important question!" Once more acknowledging her mistakes.

"Yes, you were," Betty agreed, the bitterness quite apparent.

"You didn't like me at all for it." Mirroring Betty's feelings.

"No, Mother, I didn't. If you'd told me things straight out, you could have saved me from a lot of worry and fretting . . ."

"I realize it, darling. But believe me, I didn't know enough to then."

"But you're doing all right now!" Betty's smile was radiant. "And it's twice as hard, I know, when you've had parents as old-fashioned as yours."

Then with rapport reestablished Betty confided, "I still have a lot to get straight. The basic facts--those I've managed to pick up. But there are so many details I can't figure out . . ."

As Betty went on, Mother found herself listening and answering with fewer qualms than ever before.

When it comes to you and your child, if, after thinking everything over, you still feel you don't want to, or can't, go into such discussions, if you feel that you would be too uncomfortable in coping with such questions--don't drive yourself.

You can possibly find someone else for your child to talk with. A doctor. A psychiatrist. A minister. (There is a whole group of pastors these days interested in psychology.) A family counselor. A psychologist. A gifted teacher.

Or, if you choose, you can go to someone for yourself. To help you revamp your own attitudes. You can, for example, join a group where parents can discuss their own feelings under trained leadership in an endeavor to become easier and more comfortable inside.

However, there may still be a hurdle, not in you but in your child.

Even though you get past your own old prejudices, it still may be hard for your child to discuss these matters with you. His feelings may get in the way more than with a person about whom he cares less. For one thing, he will perhaps feel less concerned with a more distant person's opinions of him should he reveal thoughts he has believed were "bad." For another thing, he may find it easier to talk to an outsider since many of his sexual worries and embarrassments arose originally from imaginings that were born out of his relationship with you, his parents, in bygone days.

Teenager - They want plain talk

Without thinking, Betty's mother had slipped back to the baby term "tee-tee" that Betty had used in bygone days. Talk of this part of the body had since then faded out of the picture. Betty had never acquired any other term. And so, when Betty's mother referred to it now, she did what came most naturally.

After listening to a speaker recently another girl in her late teens went up eagerly to the platform. "What a relief it was," she confided, "to hear you talk so simply about the things that others are so stilted about. It was good to have you use our language."

Had Betty been accustomed to the word "vulva," it would have been a different matter. As it was, the most comfortable expression for both Betty and her mother was the baby word. And so Mother's use of it in the beginning facilitated their talk. Later she was able to work in as synonyms the more scientific names.

Not infrequently language difficulties get in between parent and child and slow down communication.

Long before adolescence most children have talked with other children and have acquired their own vocabulary. By the time they are in their teens, these youngsters ordinarily have used colloquial terms among themselves. This is the language in which their imaginings and wonderings are set. As one boy put it, "We want to ask about things in the way we think about them to ourselves."

But if by chance some of these words slip out, parents are prone to grow indignant.

"P-lease, Bud, stop that gutter talk . . ."

"Heavens! Lucille! Where on earth did you hear such horrible words?"

The adolescent feels taken aback. Here is an extra stumbling block thrown in his path when he is trying so hard to find his way among conflicting ideas, supposed truths, superstitions, facts, and feelings.

He craves the familiarity of whatever language is most familiar. He likes also to feel that we are using language that is comfortable to us and that comes out of the homeyness of intimate usage, not out of a textbook lecture prepared just for this occasion and as distant as the moon and the stars.

For years in books about child psychology and sex education, parents have been told to use a scientific vocabulary. But with added observation and more vivid and direct contact with children's actions and fantasies, the eyes and ears of those who work most searchingly with them have become more perceiving.

All of us know that the mechanics of translating a foreign tongue can interfere with an understanding of what is said. Curiosity then goes unanswered. This is often what happens when we reply to our children's questions in words that are foreign and often strained. We answer technically but we leave their curiosity hanging in mid-air.

We adults very commonly use some of the same common terms as our children. And yet we avoid these terms assiduously when we talk with them.

Perhaps we should ask ourselves: Are we afraid of revealing dark secrets and shames? We needn't be. After all, gutter meanings do not lie in the words of themselves but only in whatever gutter thoughts we connect with them.

Our own use of these words has probably not been gutter usage. We have probably spoken them in moments of hearty and wholesome love talk and love play.

Without realizing it, however, by avoidance or indiscriminate condemnation, we may put needless barriers in the way of our children's free questioning and of our making things clear. We may once more give a youngster the impression that his terms are not nice and that he is not nice.

We can gain confidence better by saying, "There are lots of words for these." Then we can go on and name several of them and by so doing show that the lowly words he has most probably heard do bear mention as well as the more scientific synonyms. "I've called it by such and such term--what have you called it?" Or "These are my words for it--what are yours?" Or "Here are some of the words I've heard used. How about you?"

If we can show him that we are interested in what he has been calling things and are willing to listen, he may bring out his wonderings more readily since he can express them in his own vernacular. This may be distinctly personal. Or it may be the group language used in his particular set or locale.

We may fear, however, that if he uses these words with us he will grow too free with them elsewhere and fling them about.

But judging where and when to use certain terms is not new business in his life. Nor is it new business in our dealings with him. 'Way back when he was little we shushed him in company when he spoke about toileting. More recently, when we started applying the newer ways of discipline, we talked with him about confining his criticisms of us and his gripes against us to the privacy of our company alone. "Lots of people misjudge you when you talk in public, for instance, about being angry. They think children should always be respectful, never mad at their parents. And they think mad-talk exceedingly bad. When we're alone it's all right; but not in company."

The same sort of thing applies now. Said Randy's father, "Lots of people misjudge you when you talk like this. Many grownups believe these words are dirty. As you know, many children do too. They do have dirty meanings. But some of them have love meanings also. It all depends on how you use them. . . . I'd be careful though, because you don't want to be branded foulmouthed by people, young or old, who consider them foul."

Randy looked thoughtful and answered, "You know, Dad, you said an awful lot then. It's strange but I can usually tell whether it's the dirty use or the clean use that a person intends."

"And sometimes you want to ride along with the dirty use as well as the clean one?"

"That's true. It seems sort of smart."

At least it fits in with the revolt need that many an adolescent feels intensely. When he uses these terms smuttily, this frequently serves as outlet for hostility. Forbidding won't accomplish much. But understanding may. The terms are then no longer expressly forbidden. His use of them is no longer a gesture of throwing his parents' veto to the winds.

Healing old hurts

Betty's mother here has done something tremendously important. She has put good body feelings in the category of being good. She has lifted from them the old onus of being wicked and "bad." She has attached them to the warm and lovely yet primitive act of mothering in which body and spirit combine. And she had admitted to wishing that she might have got more of these feelings herself.

She smiled back at Betty. Then she went on. "I've learned that lots of things feel good to babies and children and to big people, too. Things we've been told we should be ashamed of. I've found out we shouldn't be at all!"

Betty stared at Mother, her eyes round and big. "What do you mean?" with a catch in her voice, eager yet somewhat afraid.

"I, too, felt my heart getting a little poundy," her mother confessed when she told of the experience. "But I decided I wasn't going to let Betty go on as I had--so stupidly ashamed of everything human. So I barged ahead! I tried to remember all the mistakes I had made in needlessly stopping Betty from enjoying her body. At the time, of course, I thought I'd been right. But why let old mistakes stay uncorrected? Especially when something as vital to your child is at stake."

Betty's mother thought back. "When you were little," she said, "I was a thumb snatcher, for one thing. I've learned now that it doesn't hurt jaws or mouth for a small child to suck. But I did everything to you then to make you stop.

"And then I was a dessert holder-upper. Custards and apple sauce and the other nourishing, sweetish foods your little body enjoyed with its taste buds! 'No,' I'd say. You couldn't have them unless you ate all the things you didn't like first. That was foolish too.

"But worst of all," and here Betty's mother drew a deep breath, "when you began to touch your tee-tee, which is the thing that feels best of all, why I had fifty-nine cat fits. I've found out since that it's harmless and I could have saved both of us a lot of worry. But I didn't know then . . ."

"You mean"--Betty's mouth hung open--"it's not true that it drives people crazy? Nothing terrible happens to them?"

"No. Not one single terrible thing. It's perfectly natural and normal and I was a very foolish woman ever to have made you think anything else."

In this, Betty's mother was bringing out what has been well established: We know now that masturbation does not injure a person or make him "nervous." We know, however, that anxiety, fear and shame over it often does.

Many an adolescent has acquired just such feelings! They intrude into the respect he has for himself as a person. And this is often what hurts him most.

At the stage where the adolescent now finds himself, sex impulses, as we know, surge freshly. The sight of a girl or a boy to a member of the opposite sex, the sound of a voice, the thrill of a chance touch, daydreams of love and romance--all bring body sensations akin to the earlier ones that were "bad." Just as the love-rivalry "bad" feelings and the touching "bad" feelings got attached to each other earlier, so the adolescent love surgings get attached now. Like a snail pulling in its horns, the boy or girl may retreat from healthy contacts. Or he may run wild to prove to himself that anxiety has no foundation.

What Betty's mother was doing was trying to reassure Betty that fear and anxiety and shame of body enjoyment had no place.

Teenage asks about birth

Knowledge about the "third opening" is relieving also when it comes to the worries over birth.

"I couldn't see how the baby was going to get out of either of the other two holes," young teenage Betty confides. "It makes a difference, though, knowing you've got such a stretchable place."

Her mother nods, "I used to think I'd burst when I had a child. Before I knew there was an opening that could stretch wide enough to let the baby through . . ."

"I used to think . . .

"I used to think . . ."

Betty chatters on, and Mother listens, adding just enough about her own former childish ideas to let Betty feel that she herself was not strange, different or "bad" because of what she had felt and thought.

"You can help lay the ghost of these earlier fantasies by talking about them," Betty's mother had learned.

"It must hurt all the same, Mom. Doesn't it--just terribly --when the baby gets born?"

"It used to, in our grandmother's day. It still did in mine. And I guess in some instances it does now. But doctors have developed new methods of pain killing. And there are ways, too, of exercising and preparing one's muscles during pregnancy that make it possible to have what's called 'painless' or 'natural childbirth.' There are books about it one can get."

"Before I have a baby, I will."

Then, after a moment, "Mother, . . . tell me . . . something else I've wondered about for the longest time. Doesn't it bother you to have a baby pulling at your breasts when it feeds?"

"I used to think it would, so I didn't nurse you children. But I wish now I had. They say it feels good."

"You mean to the mother?"

"Yes. Sort of thrilly and cozy and warm."

Betty glanced toward her mother, her face illumined. This was something new and wonderful, having mother talk about good feelings that could come to one's body. She sighed contentedly, "Gee, Mother! And I'll bet it feels good to the baby too."

Menstruation

One mother explained to her daughter Nora, whom she noticed maturing, that when she got old enough to manufacture full-grown eggs which could grow into babies she would be a woman. "You'll be able to tell when it happens . . ."

"How?"

"Some blood comes out."

"You mean that's the signal?"

"Yes. It's a kind of signal that tells a girl she's growing up."

Came the day when Nora started to menstruate. She ran into the room where her parents were reading and triumphantly announced, "What do you know, Mother and Daddy? I'm making eggs."

Actually there are egg cells inside the body when a baby girl is born. Actually these cells only become mature enough to be fertilized somewhat after the onset of menstruation. Menstruation is a sign that a girl is well on her way to this ultimate goal of womanhood. But what was more important here than these technical details was that this child was taking this physiological transition from girl to woman in good emotional stride. Many do not.

Some girls in their unconscious minds imagine they are in some mysterious fashion being punished for earlier "badness" --for curiosities that should not have been there, for littlegirl wishes to shove Mother out and possess Father or for touching themselves . . .

Threats they have heard in the past may now be catching up with them along with what they imagined when they first heard these threats.

Moans Paula, "Everytime I start flowing the thought jumps up in my mind, 'See, I've been hurt.'"

"That may be my fault," says Paula's mother. "I used to warn you not to touch yourself for fear you'd make yourself sore. I was full of those awful scare stories my mother told me. But I've learned lately that they don't hold an ounce of truth."

Some girls do not feel ready to grow up. They may unconsciously wish still to be babies. Or they may hope that by staying babyish they will manage still to get some of the things they have wanted and missed.

Cramps and nausea, aches and sick feelings may be signs of hidden unhappiness about turning from child into woman. Neither the coddling nor the belittling treatment does any good. Listening to the girl and giving her opportunities to talk about herself, on the other hand, may.

Idabelle complained every month. Her mother, before she knew better, would bring on hot drinks and hot bottles. She would dismiss her daughter's moans and groans with supposedly encouraging phrases. "You'll be all right in a few hours. And anyway, dear, this is nothing! Just wait till you have a baby. That really is grim."

Finally, however, Idabelle's mother learned better. She learned about helping a child get out troubled feelings.

"If your child feels mean, give him a chance to get the mean feelings out. That can reduce them. Talking's a good way for the adolescent youngster . . .

"If your child is afraid, give him a chance to get the fear feelings out. . . Don't probe. But don't turn him off either. Let him complain and see what comes.

"Be interested in his troubles and worries. Maybe along with the griping some of the more bothersome thoughts will slip out. If not, your willingness to listen may of itself do more than you think."

So Idabelle's mother figured, "I'll give it a try."

"Oh," groaned Idabelle with a sour look on her face. "I feel so awful!"

"I know how it is," said her mother, drawing up a chair and settling back to listen, real interest showing.

"I hurt so in my middle!" Idabelle moaned. "I wish I'd feel better. I just can't miss school . . . I can't miss the tennis tournament . . ." She paused a few moments and then, very angry, she exclaimed, "Why on earth did you make me be born a woman? I'd have avoided all this misery if you'd only let me be a boy!"

Idabelle raved on until, as suddenly as the complaining had started, it stopped. Idabelle burst out laughing. "Gee, Mom, did you ever hear anything half so absurd?"

"Yes, I have," Mother answered. "Me! I've painted a woman's life in pretty grim colors, as if men had all the advantages . . ."

"Well, they have, haven't they? Look at Jack. He never gets this 'curse.' He goes more places with Dad. He plays tennis twice as easily. Beside which I think you've always preferred him . . ."

"It's true that mothers do have a special kind of feeling toward their sons. But they also love their daughters in a different way."

That was all for the moment. However, with additional opportunities for voicing the fantasies that she would have been more beloved and happier if she'd been a boy--Idabelle grew less incensed about her "horrible fate," and her pain eased.

Even though no "revelation" ever rolls out, acceptant listening that communicates understanding can sometimes of itself bring more relief than do pills.

Many parents believe it wise to check with the doctor if a girl continues to have trouble with her periods. Seldom does the doctor find anything physically wrong. But he can sometimes step in and give the girl opportunities to talk in a more casual and easy way than the parent, who is naturally more emotionally involved. Only under very rare circumstances and only if the doctor finds unusual and severe symptoms should there be actual examination of the sex organs. For the young girl starting out on her course of being a woman such an examination more frequently than not stands as a very grave and shocking threat, as a punishment for secret thoughts, as an invasion of privacy and as a violation of her whole person.

Irregularity in menstruation is very common. Sometimes girls skip several months in between each of their first few periods. Most women remain somewhat irregular and can expect at least a third of their cycles to extend past the time they believe it is due. The idea that one has to be regular to be normal sets up tension which, in its turn, like other worries, may cause delay.

"I'm late," wails young Jinny. "And I know my mother'll think the worst. She never trusts Bob and me. She watches me like a hawk, expecting me to skip . . ."

The worry made young Jinny late just as an intense desire to be pregnant has made many a woman late, or the intense desire not to be pregnant. Letting down on tension often does wonders. Talking out worries to an acceptant listener may help to bring this about.

Prior to the onset of their menses, many girls are worried about personal hygiene and want information concerning it. Showing them, if they wish, just how to wear their napkins instead of merely telling them can make for a bit of needed assurance. However, insisting on such demonstration if they wish to avoid it becomes an invasion of the privacy they crave.

Discussing the matter of bathing and of exercising is also in order. "Many girls go swimming, bathe, take showers, play tennis and enter into other games as usual. Many girls prefer not to do strenuous things. Ordinarily, it depends on psychological choice rather than on physical necessity . . ."

Boys are fully as curious, and often as worried, about menstruation as are girls.

"But why on earth are they so callous?" one mother complained. "When my daughter has her period and doesn't feel like swimming, her brother starts in on her: 'What's the matter?' . . . 'What's happened to you?' . . . 'Why don't you get in the water? You're chicken! What's wrong?' . . . I've told him girls have what's called their period once a month and during that time he should be more considerate. But I might as well talk to a lizard for all the attention he pays."

Wednesday, October 10, 2007

Preretirement Planning

One of the most important contributions to a successful retirement is advance planning well ahead of the actual date of retirement. Where you will live, and what your continuing activities will be are questions that should be considered. A location convenient to your children can be a great asset both to them and to you. Many retirees seek a year-round warm climate, while others prefer to remain where their roots are established.In developing financial plans for retirement, it is helpful to outline personal needs and preferences. Although individual circumstances and goals may differ, the following are general guidelines:

1. An income payable each month of retirement during your lifetime sufficient to enable you to maintain the same standard of living you and your spouse enjoyed before you retired. Because of tax reductions and a different pattern of expenses in retirement, the gross income needed may be only 80 percent or less of your preretirement income. However, it will be necessary to build protection against the ravages of inflation that decrease the value of the dollar.
2. Continuing income payable to your spouse after your death sufficient to maintain your customary standard of living.
3. Hospital-Medical-Surgical coverage which, combined with Medicare, will enable you to meet the major share of medical expenses.
4. As large a nest egg as possible for special needs, such as travel or unforeseen expenses. The nest egg may include a savings account, investments in money-market type mutual funds, bonds, or stocks, the equity in your home, and the L cash values of your life insurance contracts.

Careful advance financial planning can also reduce later retirement expenses, for example, completing the mortgage payments on your home before you retire. Plan to enter retirement with a new car or one of recent vintage in good condition. Consider putting your life insurance on a paid-up basis at retirement to avoid further payment of premiums. Plan to purchase desired high-cost items before retirement.

Because it is not possible to predict how long you will live in retirement, you should prepare for a wide range of possibilities: a long life, a short one, or something in between.

The Teachers Insurance and Annuity Association (TIAA), which provides pensions to the' majority of college professors, reported in 1979 that 778 of its annuitants were over 90 years old and 17 were over 100. Its oldest pensioner was 107 years old.

Clearly, long life is possible and it is well to avoid outliving your income. Social Security, annuities, and your company pension will continue until your death, and, in some cases, will continue after that in whole or in part to your named beneficiaries. But if savings are drained too rapidly, you may outlive them.

The following table shows the number of additional years people live on the average after reaching a given age. However, these are only, averages not a basis for firm planning. Many will not live as long as indicated; others will live much longer. As medical knowledge advances, these average expectancies may increase.

Average Additional Years of Life

Age Male Female
55 23 28
60 19 23
65 15 19
70 12 16
75 9 12
80 7 10
85 5 7
90 4 6
95 3 4
100 2 3

Thursday, September 27, 2007

Biotin

The term "biotin" should now replace several old names, to wit: coenzyme R, protective factor X, the protective factor against egg white injury, the vitamin H and a part of the Bios II b or the adsorbable factor of the yeast growth promoting substance. It was isolated and from the Bios II b factor of yeast as a methyl ester of the empirical composition C 11 H 18 O 3 N 2 S and having a melting point of 166°-167° when highly purified. Saponification with cold alkali gives free biotin of the empirical composition C 10 H 16 O 3 N 2 S and melting at 230°-232°. Biotin is a simple monocarboxylic acid, a derivative of valeric acid. The nitrogen atoms form a urea structure which can be opened up with a loss of one carbon to form a diaminocarboxylic acid, from which in turn one can reform biotin by reaction with phosgene. The sulfur is functioning in a thioether structure, since a sulfone can be formed by oxidation. The most probable structure is that of 2′-keto-3,4-imidazolido-2-tetrahydrothiophene-n-valeric acid.

Looked upon as an active principle of yeast growth biotin is active even at a concentration of one part in 500,000,000,000. As the active substance stimulating several species of the root-nodule bacterium Rhizobium and obtained from concentrated cultures of Azotobacter, and previously known as coenzyme R (R=respiration) it shows effects in concentration of I part in 100 billion. As Vitamin H (H for "Haut" or skin) it counteracts the effects of avidin of both raw and dried egg white to the extent of 10,000 units per milligram by a rat assay method (i.e. 0.1 gamma of biotin per rat per day for 30 days protects against egg-white injury).

The biotin may be extracted from liver, dried eggs, potato starch, fresh eggs, dried yeast, autolyzed yeast.Inactivation of biotin has been accomplished both under acid and alkaline conditions. Aeration is not effective but stronger oxidizing agents destroy activity quickly and completely. Nitrous acid inactivates without a loss of nitrogen.Biological effects of biotin other than those used in assay of the foregoing effects concern the fatty infiltration of the livers of rats on a low fat diet with high biotin. The fermentation of yeast is increased as well as its respiration and growth both aerobically and anaerobically, in the presence of plentiful nitrogen in the form of ammonia. Butter-yellow tumor formation brought on by avidin-containing diets should be counteracted by biotin.

Biophysics scope principles methods

The domain of discussion may be indicated by defining biophysics as the application to biological research of the methods and content of the physico-mathematical sciences. "Method" is advisedly placed first. It has become clear that the experimental and theoretical procedures usually associated with physics are not the exclusive property of that science, but represent rather adequately the developed and mature form of the scientific method itself. Biophysics is the adaptation of this methodology to the problems of biology.

It appears from this that the word "biophysics" is in part a misnomer. Biophysical research is biological research with a particular emphasis--concern for the logical ordering and quantification of biological theory, for the more intimate unification of theory and experiment, for the introduction into experimental biology of techniques and measuring devices which have been developed in the so-called physical sciences, and for the reduction-when it is possible and useful, and only then-of biological problems and concepts to already existent laws and concepts of the physical sciences.

That physics--the science of matter--has in fact preoccupied itself with those objects which used to be termed "non-living" is logically accidental, not intrinsic. Taken in this sense, the "physicalizing" of biology merely means the attempt to analyze certain complex phenomenal patterns in terms of somewhat simpler patterns which have already been the object of intensive study. In this sense it is neither novelty nor heresy, but an inevitable extension of the classical biological procedure of analyzing organisms into organs, organs into tissues, tissues into cells, and cells finally into their discernible parts.

It is only fair to add that "biophysics" is usually used in a sense which contrasts it with biochemistry--that is, as a study of the physical as distinct from the chemical aspects of biological systems. This viewpoint would see viscosity, osmotic pressure, surface tension, the electrical and mechanical properties of biological systems as physical, while the composition and metabolic activities of such systems are the concern of chemistry.

This criterion has been serviceable in the past, of course; but it is so narrow as to be both cramping to research and rather difficult rigidly to maintain. Thus, the study of bioluminescence must in this view separate sharply into two parts: the analysis of the luciferinluciferase system is biochemistry, while the emission of a photon by the reduced enzyme is biophysics. It is valid to say, "For the moment, let us consider only the physical aspects of this phenomenon, and ignore the chemical and purely biological." But the moment passes quickly, and the essential inseparability of the different aspects demands the broader interpretation.

It remains to consider the best way of subdividing biophysical activity into its branches according to some specified criterion of classification. From the preceding discussion it is clear that one apparently natural division-into theoretical and experimental--is not a desirable one. The nature of biophysics, as here defined, demands the same close interweaving, mutual support, and mutual stimulus of theorizing and experimenting as is now practised so successfully in physics.

To adopt the existing fundamentum divisionis of biology is no more desirable; for the fences around bacteriology, embryology, histology, physiology, botany, and the like, in view of their haphazard historical origin, exhibit small regard either for practical utility or for logical consistency. Some of these fields are organized around a particular function, some around a particular set of structures, some around a particular class of organisms, and some are mere catch-alls for topics which do not fit handily into the remaining cells.

If we accept the cell as fundamental concept in biology, a useful though by no means perfect mode of classification suggests itself--the hierarchy: cell-parts, cells, cell-aggregates. These further subdivide in a fairly natural way, principally according to functions and activities. Thus, under Cell-parts, we think of: Protoplasmic Structure; Enzyme Systems of the Cell; Nucleus and Chromosomes; Cell Membranes and Permeability; Golgi Bodies and Mitochondria.
Under Cells would come: Growth; Form; Motion; Division; Differentiation; Metabolism; Senescence; Stimulus-Response.

Under Cell-aggregates there are similar categories: Organic Form and Differentiation; Growth; Metabolism; Senescence; StimulusResponse. These would intersect with the triple classification: Tissues; Organs; Organisms. Thus, for example, we would find Secretion under Metabolism, Organs (perhaps also under Cells, Metabolism); special senses under Stimulus-response, Organs; and the nervous and endocrine systems under Stimulus-response, Organisms. Another category under Cell-aggregates would be Populations, which would cover the material of biometrics, ecology, and much of bacteriology.

Such a topic as The Virus is not an insuperable obstacle to this arrangement; there are many good reasons for placing it under CellParts. But it is obvious from the topics named that a better method than either equal-ranking classes or a hierarchy would be a multidimensional network, in which topics may be different distances apart, but in which each topic is linked to many others directly, and to all others indirectly. Protoplasmic Structure would be close to if not precisely at the centre of such network. With the further development of biophysics, the appropriate ordering will doubtless become more evident.

Applied biophysics is exceedingly scanty. There are a few scattered medical applications, such as the gold number or its modern equivalents in the analysis of cerebrospinal fluid, the use of X-rays and radium in the treatment of cancer. But biophysics is yet too young to show applications on any large scale.

There are many fields of biology in which the rather young biophysical method has not been active, so that a sufficient amount of representative material is not available for presentation. Some topics of biophysical interest, e.g., metabolism, are still referred to biochemistry for the same reason.

Bioluminescence

Bioluminescence is a word applied to the light emitted by various living organisms. At least 40 different orders of animals contain luminous species and two groups of plants, the bacteria, responsible for the luminescence of flesh and dead fish, and the fungi, which live in phosphorescent wood. Luminous bacteria are so small that individuals cannot be seen by their own light but colonies are visible. They are easily cultured and are nonpathogenic to man but may infect living animals, giving rise to a luminescent disease of sand-fleas, shrimps and midges, which is eventually fatal. Luminous bacteria may also live symbiotically in special organs of certain fish, notably Photoblepharon and Anomalops, of the Banda islands.

Bacteria and fungi emit light continuously day and night, while all other forms luminescence only when stimulated. The phosphorescence of the sea appears when many different kinds of small organisms are disturbed by the breaking of waves or motion of a boat. Among the groups of animals containing luminous species are flagellates, radiolaria, sponges, jelly-fish hydroids, sea pens, ctenophores, nemerteans, earthworms and many marine worms, shrimp, ostracods and copepods, myriapods, several groups of insects, monuses, squid, brittle stars, balanoglossids, tunicates and fish.

Bioluminescence is never dependent on a previous illumination of the cells or a previous radiation of any kind, nor is it connected with crystallization, friction, or rubbing, but is the result of oxidation by molecular oxygen of a definite substance produced in the luminous cell. It is a chemiluminescence. The luminous material or photogen is almost universally manufactured by living cells as granules, which may normally undergo oxidation within the cell, as in the fire-fly, (intracellular luminescence) or be extruded as a luminous slime or secretion (extracellular luminescence) as in a small ostracod crustacean, Cypridina. Most is known concerning the chemistry of extracellular luminescence, especially that of Cypridina.

In Cypridina the granules in the secretion dissolve on contact with sea water and the homogeneous luminescence is emitted by the resultant colloidal solution. Two kinds of granules are distinguishable in the cells of Cypridina, one large and yellow, the other small and colorless. In fact, in five of twenty-five different groups of luminous animals tested, it can be demonstrated that luminescence is due to two chemical substances, luciferin (yellow) and luciferase (colorless), which can be easily separated because of a difference in resistance to heating and other properties.

Crude luciferin solution is prepared by making a hot water extract of a luminous organ. Heating destroys the luciferase but does not harm the luciferin. Crude luciferase solution is prepared by making a cold water extract of a luminous organ, when both luciferin and luciferase dissolve and luminescence occurs. The extract is then allowed to stand in the air until the light disappears, evidence that the luciferin has been completely oxidized, leaving the luciferase, an enzyme, in solution. A luciferase solution, by virtue of this mode of preparation, must contain the oxidation product of luciferin as well as luciferase.

Luciferin and luciferase are quite specific. Luciferin from one animal will not luminesce if mixed with the luciferase of another luminous form unless the animals are closely related, such as two species of the same genera or two genera within the same order. Even if the color of the luminescence is different in the two different species light will appear, provided the species are closely related. In this case it is interesting to note that the color of luminescence of the resultant "cross" is determined by the animal supplying the luciferase. Luciferase must be the source of the light. It is convenient to designate the luciferins and luciferases by prefixing the name of the animal from which the substances are obtained.

Most luminous animals, if dried rapidly will again luminesce on moistening. Dried Cypridinae have been kept for 26 years without deterioration and can be used for preparing luciferin and luciferase.

Cypridina luciferin is purified by extraction of the dry Cypridinae with methyl alcohol. Ten per cent of butyl alcohol is then added and the methyl alcohol removed in vacuo. The supernatant butyl alcohol extract is chilled and benzoylated with benzoyl chloride. After fifteen minutes this solution is diluted with ten volumes of water and the new inactive benzoyl luciferin derivatives extracted with pure ether. After removing the ether in vacuo the residual liquor is hydrolyzed with hydrochloric acid in absence of oxygen. The free active luciferin is left in the acid solution and can be extracted with butyl alcohol. By repeating the benzoylation and hydrolysis, a product purified 2,000-fold, as compared with dry Cypridinae, can be obtained.

To purify luciferase it is usually sufficient to dialyze a cold, wellstirred water extract of rapidly dried, powdered Cypridinae against cold running water for twenty hours. Dialysis removes pigment, and a precipitate forms which can be filtered off. A few drops of toluene added to this solution will preserve it for months with little loss in activity if kept in a refrigerator.
Cypridina luciferin is slowly dialyzable; not destroyed by trypsin; soluble in water, absolute methyl, ethyl, and propyl alcohol but insoluble in acetone, benzene and ether; readily adsorbed on fine particles. It does not act as an antigen.

Cypridina luciferase is non-dialyzable; destroyed by trypsin; soluble in water, but insoluble in alcohols and all fat solvents; readily adsorbed on surfaces. It is capable of forming an antiluciferase when injected into the blood of a rabbit. Other luciferins and luciferases have different properties.

The oxidation of luciferin with production of light in the presence of luciferase gives an oxidation product which cannot be reduced, whereas the oxidation without luminescence by oxidants like potassium ferricyanide is reversible.

This Product is called oxidized lucifenn. The change with ferricyanide occurs in two steps, one of which is the reversible oxidation previously referred to; the second is irreversible and probably also an oxidation, although this has not been definitely demonstrated. The spontaneous oxidation of luciferin without emission of light in crude solutions (without luciferase) is probably catalyzed by traces of heavy metals in the solution and proceeds much more slowly when the luciferin has been purified. Both the non-luminescent oxidation and the luminescent oxidation undoubtedly take place simultaneously when luciferin is mixed with luciferase.

Cyanide does not affect luciferase but forms an irreversible combination with purified Cypridina luciferin. Azide combines reversibly with luciferin whereas urethane, sulfanilamide, sulphathiazol, sulphapyridine and p-aminobenzoic acid probably act reversibly to inhibit luciferase activity.

Divisions of biochemistry

No rigid, conventional divisions of biochemistry exist, as in the case of chemistry and physics proper, where a distinction between organic' and "inorganic," for example, is rather sharp. Still dominant is the treatment where the class of substance involved is the name of the subject, as "Carbohydrates," "Fats," "Proteins. . . . . Inorganic Constituents," "Accessory Substances," and now '"Hormones." Almost all texts describe these groups of substances and then take up what is known of their ingestion or synthesis in the body, their role, their metabolism and the further course, final consumption and elimination of the products of their metabolism. That at least the principal classes of materials serve as equivalents of one another and are mutually convertible tends to upset any such classification. The latter part of many a text, therefore, resorts to a series of chapters where the physiological process, e.g. digestion, or the function of an important organ, e.g., the kidney or the liver, serves as the division of the subject.

Occasionally a text starts with a phsico-chemical introduction in place of the organic chemistry of the principal materials. In such cases a stress on colloid chemistry is imperative. The fields of biochemistry shape up as applications of sectors of the other sciences to living systems.