HEMOGLOBIN PRODUCTION FACTORS IN THE HUMAN LIVER : III. ANEMIAS—PRIMARY, APLASTIC AND SECONDARY—LEUKEMIAS

Biological assay of the human liver in various types of anemia shows conspicuous differences in the concentration of hemoglobin producing factors. Pernicious anemia shows very high values and the liver in untreated cases may show maximal storage of the hemoglobin producing factors. Liver therapy reduces this store as the missing factor is supplied and new hemoglobin and red cells can be turned out by the marrow. Aplastic anemia likewise shows high concentration of hemoglobin producing factors as there is no outlet for this material through the red marrow. Secondary anemia due to loss of blood will show low normal values but even long standing severe anemia will not seriously deplete this store of hemoglobin producing factors in the liver. Secondary anemia due to blood destruction within the body shows higher values and some excess store of hemoglobin producing factors and iron. Leukemia gives a biological assay like secondary anemia due to blood loss and always presents definite anemia. Iron analyses show conspicuous differences and iron concentration within the liver parenchyma does not in any way parallel the concentration of hemoglobin producing factors. The highest values for iron concentration are found in aplastic anemia (70 mg. per cent)— high values in pernicious anemia (51 mg. per cent)—normal values in leukemia (13 mg. per cent)—and low values in anemia due to loss of blood (5.3 mg. per cent). These findings should aid in a more complete understanding of the pathogenesis and internal metabolism of various anemias

HEMOGLOBIN PRODUCTION FACTORS IN THE HUMAN LIVER : ANEMIAS, HYPOPROTEINEMIA, CIRRHOSIS, PIGMENT ABNORMALITIES, AND PREGANCY

Human liver tissue has been assayed to determine the amount of hemoglobin production factors in normal and abnormal states. Standardized dogs made anemic by blood removal have been used in this biological assay. Normal animal liver as control is rated as 100 per cent. Normal human liver tissue as compared with the normal animal control contains more of these hemoglobin production factors—a biological assay ratio of 120 to 160 per cent. Infections, acute and chronic, do not appear to modify these values, the concentration of hemoglobin-producing factors falling within the normal range. Pernicious anemia and aplastic anemia both show large liver stores of hemoglobin-producing factors—a biological assay ratio of 200 to 240 per cent. Therapy in pernicious anemia reduces these liver stores as new red cells are formed. Secondary anemia presents a low normal or subnormal liver store of hemoglobin-producing factors—an assay of 60 to 130 per cent. Hemochromatosis, erythroblastic anemia, and hemolytic icterus in spite of large iron deposits in the liver usually show a biological assay which is normal or close to normal. Polycythemia shows low reserve stores of hemoglobin-producing factors. Leukemias present a wide range of values discussed above. Hypoproteinemia almost always is associated with low reserve stores of hemoglobin-producing factors in the liver—biological assays of 60 to 80 per cent. Hypoproteinemia means a depletion of body protein reserve stores including the labile protein liver reserves—a strong indication that the prehemoglobin material (or globin) is related to these liver stores. Pregnancy, eclampsia, and lactation all may present subnormal liver stores of hemoglobin-producing factors. Exhaustion of protein stores lowers the barrier to infection and renders the liver very susceptible to many toxic substances. It should not be difficult to correct hypoproteinemia under these conditions and thus relieve the patient of a real hazard

DIETARY EFFECTS ON ANEMIA PLUS HYPOPROTEINEMIA IN DOGS : II. THE FINDINGS WITH MILK PRODUCTS, WHEAT, AND PEANUT FLOURS AS COMPARED WITH LIVER

Casein (purified or commercial) in this type of experiment falls in the top bracket as a protein consistently favorable for maximal new hemoglobin and plasma protein production in doubly depleted dogs (anemic and hypo-proteinemic). Lactalbumin is less favorable for total blood protein production and the ratio of plasma protein to hemoglobin is high—that is lactalbumin favors plasma protein production as compared with casein, or is less favorable for hemoglobin production. Peanut flour (purified or commercial) is less than half as effective as casein in promoting new blood protein production. The ratio of plasma protein to hemoglobin is about the same as casein. Wheat gluten as tested is distasteful to dogs. It is neither very good nor very poor for blood protein production when it is eaten. There is nothing unusual about the response. Weight loss usually confuses the picture. Liver stands as a control base line for the above experiments. Its capacity to further hemoglobin and plasma protein production is well established. The production of hemoglobin was about 3 times that of plasma protein in the experiments

COPPER AND COBALT RELATED HEMOGLOBIN PRODUCTION IN EXPERIMENTAL ANEMIA

Copper added to a standard diet often effects a moderate increase in hemoglobin production in anemia due to blood loss. The copper response is quite irregular in contrast to the iron response. In these dogs there is no lack of copper held in reserve stores (liver and spleen) so the reaction is not related to an actual deficiency of the element. An effect upon enzyme complexes related to globin and hemoglobin production is to be considered. Cobalt under similar conditions causes no stimulus to hemoglobin production, rather an inhibitory effect when more than minimal doses are given. The claim that cobalt causes a polycythemia in dogs receives no support from our experiments

GLOBIN UTILIZATION BY THE ANEMIC DOG TO FORM NEW HEMOGLOBIN

It has been shown that the standard anemic dog can use sheep, goose or dog hemoglobin when given by vein and return quantitatively its equivalent as new dog hemoglobin within the red cells. Globin at times can be used when given by vein with a quantitative return of new hemoglobin in red cells in these same anemic dogs. Again the administration of globin by vein will inhibit the expected hemoglobin formation; and we believe the toxic effect of the globin is responsible. A digest of globin may be used by the anemic dog to form new hemoglobin. Globin from both horse and dog have been tested and seem to react in identical fashion. The globin radicle of hemoglobin appears to be an important limiting factor in abundant hemoglobin building in this type of anemia due to blood loss. Globin fed by mouth is well utilized to form new hemoglobin and we may record a 30 to 40 per cent utilization or a return of 30 to 40 gm. new hemoglobin from the feeding of 100 gm. globin. This is to be compared with the utilization of liver protein—an average return of 13 gm. new hemoglobin for the feeding of 100 gm. liver protein

HEMOGLOBIN PRODUCTION FACTORS IN THE HUMAN LIVER : I. NORMAL, INFECTION AND INTOXICATION

Human liver tissue has been assayed to determine the concentration of hemoglobin production factors in normal and abnormal states. Standardized dogs made anemic by bleeding have been used in this biological assay and the human liver tissue compared with control animal tissue. Normal human liver tissue (external trauma) contains much more of these hemoglobin production factors than the normal control animal liver—the ratio being 162 to 100. In this form of biological assay 42 gm. of animal liver or 26 gm. of human liver represent 1 gm. of potential hemoglobin. A second group (Table 2) in which the viscera were practically normal except for atrophy, the cases presenting a good deal of arteriosclerosis and senile changes, shows a lower content of these hemoglobin production factors. The ratio of human to control here is 117 to 100. This is certainly the low limit of normal. Acute fulminant infections reduce somewhat the store of these potent hemoglobin production factors in the human liver (Table 3). The average value is 117 as compared with 100 control but the more acute cases show the lower values. Chronic intoxications show values which are close to the human normal—151 per cent. The liver content of hemoglobin producing factors shows very wide fluctuations in cases of thyrotoxicosis. Diabetes may be associated with rather low values. There may be complete dissociation of the organic iron content and the concentration of hemoglobin production factors in the liver

HEMOGLOBIN PRODUCTION FACTORS IN THE HUMAN LIVER : II. LIVER DEGENERATION, CANCER, CIRRHOSIS AND HEPATIC INSUFFICIENCY

Biological assay of human liver material shows various concentrations of hemoglobin producing material in various diseased states. Cardiac congestion with red atrophy of the liver shows values (Table 21) somewhat below the human normal—94 per cent compared to 162 per cent. It is probable that this is due to atrophy and destruction of the liver cells in the centers of the liver lobules. We believe the remaining liver cells have a normal content of hemoglobin producing factors. Amyloid (tuberculosis) and fatty (alcoholic) degeneration show (Table 22) subnormal values—111 per cent compared to 162 per cent. If we allow for the "dilution" of liver cell weight by the amyloid and fat we observe normal values for the concentration of hemoglobin producing factors. Carcinoma growth in the liver whether primary or secondary shows net figures (Table 23) which are low—75 per cent compared to normal 162 per cent. It appears that cancer tissue, even tumor cells originating from hepatic epithelium, contains no hemoglobin producing factors. If we allow for the "dilution" of the remaining liver cells by the cancer tissue the figures are within normal limits. Cirrhosis and hepatitis (Table 24) with no signs of true hepatic insufficiency or severe hepatic injury but death supervening from secondary infection or hemorrhage will give average normal values –164 per cent compared to normal human of 162 per cent. Icterus is not a factor. Cirrhosis or hepatitis with severe parenchyma injury and signs of hepatic insufficiency give low values (Table 25) in biological assay –48 per cent compared to normal human 162 per cent. It may be surprising that the liver cells hold so tenaciously to these unknown factors influencing hemoglobin production but obviously only in severe liver injury is this concentration seriously reduced

ANEMIA PLUS HYPOPROTEINEMIA IN DOGS : VARIOUS PROTEINS IN DIET SHOW VARIOUS PATTERNS IN BLOOD PROTEIN PRODUCTION BEEF MUSCLE, EGG, LACTALBUMIN, FIBRIN, VISCERA, AND SUPPLEMENTS

Dogs with sustained anemia plus hypoproteinemia due to bleeding and a continuing low protein or protein-free diet containing abundant iron have been used in the present work to test food proteins and supplements as to their See PDF for Structure capacity to produce new hemoglobin and plasma proteins. The reserve stores of blood protein-producing materials are thus largely depleted in such animals and sustained levels of 6 to 8 gm. per cent hemoglobin and 4 to 5 gm. per cent plasma protein can be maintained for considerable periods of time. The stimulus of double depletion drives the body to use all protein building materials with the utmost conservation. This represents a severe biological test for food and body proteins and its assay value must have significance. Measured by this biological test in these experiments, casein stands well up among the best food proteins. The ratio of plasma protein to hemoglobin is about 40 to 50 per cent, which emphasizes the fact that these dogs produce on most diets about 2 gm. hemoglobin to 1 gm. plasma protein. The reason for this preference for hemoglobin production is obscure. The mass of circulating hemoglobin is greater even in this degree of anemia and the life cycle of hemoglobin is much longer than that of the plasma protein. Egg protein, egg albumin, and lactalbumin all favor the production of more plasma protein and less hemoglobin as compared with casein. The plasma protein to hemoglobin ratio is increased, sometimes above 100 per cent. Supplements to the above proteins of casein digests or several amino acids may return the response toward that which is standard for casein. Histidine as a supplement to egg protein increases the total blood protein output and brings the ratio of plasma protein to hemoglobin toward that of casein. Beef muscle goes to the other extreme and favors new hemoglobin production up to 4 gm. hemoglobin to 1 gm. plasma protein—a ratio of 25 per cent. The total amounts of new blood proteins are high. Lactalbumin as compared with casein shows a lower total blood protein output and a plasma protein to hemoglobin ratio of 70 to 90 per cent. Amino acid supplements are less effective. See PDF for Structure Fibrin is a good food protein in these experiments—much like casein. When fed over these 5 week periods it causes a sustained increase in blood fibrinogen. Folic acid in the doses given has no effect on the expected response to various diets. Peanut flour is a very poor diet for the production of new hemoglobin and plasma proteins. Small supplements of casein and beef show a significant response with improved output of blood proteins. Soy bean flour gives a poor response and wheat gluten a good response with adequate output of blood proteins. Visceral products show some variety. Beef heart is not as effective as beef muscle. Beef spleen, kidney, and pancreas give good responses but not up to casein. Pig stomach, beef brain, and calf thymus are below average. The plasma protein to hemoglobin ratio shows a narrow range (40 to 60 per cent) in experiments with visceral products

INFECTION AND INTOXICATION : THEIR INFLUENCE UPON HEMOGLOBIN PRODUCTION IN EXPERIMENTAL ANEMIA

Infection in human cases is often believed to be responsible for anemia. It is generally believed that lack of absorption and definite blood destruction are responsible for the anemia. Accelerated metabolism due to thyroid or dinitrophenol does not modify hemoglobin production in these standard anemic dogs. Endometritis lasting over many weeks will profoundly reduce the production of hemoglobin in the standard anemic dog. A sterile abscessalso will diminish the production of new hemoglobin in the anemic dog when liver is being fed but particularly during fasting periods when the usual abundant production of new hemoglobin is reduced to zero. Impaired absorption can be excluded as a factor of any significance in certain experiments given above. Destruction of red cells can likewise be excluded as of any significance in certain experiments given above. These experiments point to a disturbance of internal metabolism related to hemoglobin building in the body as responsible for the inhibition of hemoglobin production under these conditions. We believe this same factor is often of importance in human disease

DIETARY EFFECTS ON ANEMIA PLUS HYPOPROTEINEMIA IN DOGS : I. SOME PROTEINS FURTHER THE PRODUCTION OF HEMOGLOBIN AND OTHERS PLASMA PROTEIN PRODUCTION

Doubly depleted dogs (anemic and hypoproteinemic) respond favorably to all the diet proteins used in the above experiments. Egg products (whole egg, albumin, or egg yolk) are well utilized by these dogs. Egg proteins favor the production of plasma protein and in some experiments the output of plasma protein is actually more than the output of hemoglobin. In contrast fresh beef muscle favors hemoglobin production—the output being 3 or 4 times that of plasma protein. The processed egg albumin fed in Table 4 was not well utilized and there was weight loss. Beef muscle (fresh or processed) gives a total blood protein output about twice that with egg feeding and there is a striking preponderance of hemoglobin output. Beef heart and salmon muscle show a pattern much like beef muscle. The total blood protein output is below that due to beef muscle