The effect of quantitative and qualitative protein deficiency on blood regeneration; hemoglobin

1. The effect of diets, varying in quantity or quality of protein, on hemopoiesiswas studied in protein depleted and anemic adult rats.

2. In experiments with diets containing different amounts of casein (0, 3, 9 and18 per cent, respectively), and fed ad libitum, it was found that with a protein-freediet a further decrease of hemoglobin occurred, whereas the other three dietsinitiated a regeneration of hemoglobin; its degree being more or less proportionalto the casein content.

3. In experiments, in which diets with 9 and 18 per cent of casein, respectively,were given in restricted amounts, it was found that the degree of hemoglobinformation was similar to that with the same diets when given ad libitum, whereasthe weight gain was considerably less. It is concluded, therefore, that in caloricdeficiency hemoglobin formation has a priority over weight recovery.

4. A diet containing 30 per cent casein and given in restricted amounts induceda further weight loss, whereas the concentration of hemoglobin showed a markedincrease. Comparing the results obtained by this diet with those observed with 18per cent casein diets, given either ad libitum or in controlled amounts, it was evident that restriction of the quantity of food protein interferes more seriously withhemopoiesis than restriction of calories.

5. Diets containing nutritionally inferior proteins fed at 9 per cent level, alsoimpaired normal hemopoiesis. Hemoglobin regeneration induced by the proteinsinvestigated was found to decrease in the following order: eggs, meat, processedsoya, casein, peanut, maize, wheat, gelatin.

6. Comparing the nutritive value of various proteins for regeneration of hemoglobin and of granulocytes it was found, that casein and soya and maize proteinsare considerably more efficient for hemoglobin formation than for production ofgranulocytes, whereas wheat protein and gelatin have a higher granulocytopoieticcapacity.

     

Study of Leukopenias and Thrombocytopenias by the Direct Antiglobulin Consumption Test on Leukocytes and/or Platelets

A new test, the direct antiglobulin consumption test (direct ACT), has beendevised to be performed on the leukocytes and platelets of patients. It wasperformed in parallel with the indirect antiglobulin consumption test (indirectACT), and other serologic tests on 492 individuals comprising 24 cases of diffuse lupus erythematosus, 93 primary thrombocytopenias, 18 secondarythrombocytopenias, 48 primary leukopenias or leukothrombocytopenias, 80secondary leukopenias, 101 non-leukothrombocytopenic patients and 128 normal subjects.

A good correlation was obtained between a positive or negative result andthe presence or absence of a cytopenia in the corresponding cell series. Outof 183 thrombocytopenic patients, 42.2 per cent gave a positive result withplatelets, and out of 128 leukopenic patients, 25.8 per cent gave a positive result on leukocytes, whereas of the 299 patients with normal leukocyte andplatelet counts, 98 per cent gave negative results. The test was found positivein three categories of patients:

(1) In diffuse lupus erythematosus, tests on both leukocytes and plateletswere almost uniformly positive. The indirect ACT permits a distinctionto be made between three substances in the serum of these patients.These are antiplatelet, antileukocyte-cytoplasm and antileukocyte-nuclearsubstances.

(2) The thrombocytopenic patients were subdivided into two groups, namely primary and secondary thrombocytopenia:

(a) Out of 93 cases of idiopathic thrombocytopenic purpura, about50 per cent gave a positive result with platelets. Neither thehistory nor the clinical picture suggested any differentiation between those who gave negative results, apart from the fact thata history of infection was more frequent among the negativecases. Corticosteroid therapy did not affect the result of the test,but following splenectomy, 38 per cent of the cases become negative.

(b) Of 18 patients with a secondary thrombocytopenia, three cases(16 per cent) gave a positive result.

(3) The leukopenic and leukothrombocytopenic patients were also subdividedinto two groups:

(a) Out of 48 primary or idiopathic cases, some 50 per cent gave apositive direct ACT either on leukocytes and/or on platelets. Byusing the indirect ACT it was possible to distinguish two substances in the serum, one being antiplatelet and the other an antileukocyte cytoplasmic substance.

(b) The 80 cases of secondary leukopenia or leukothrombocytopeniagave a positive result in 15 per cent of cases with leukocytes and/or with platelets.

Of the 101 non-leukothrombocytopenic patients, only five were found togive positive tests.

All the 128 normal subjects gave negative results.

The direct ACT provides direct evidence of the presence of a -globulin,probably an auto-antibody, on the leukocytes and/or the platelets of some 99per cent of cases of diffuse lupus erythematosus, of about 50 per cent ofcases of idiopathic thrombocytopenia and idiopathic leukothromobcytopenia,and in about 15 per cent of cases of secondary thrombocytopenia and leukothrombocytopenia. No marked difference was found in the history, clinicalpicture, or hematological findings between patients giving positive and negative results.

Submitted on April 3, 1961 Accepted on August 1, 1961     

Experimental production of a nutritional macrocytic anemia in swine

1. A deficiency of pteroylglutamic acid has been produced in 32 swine fed apurified diet containing casein and supplemented with seven B vitamins, sulfasuxidine and a folic acid antagonist. The casein was fed at two levels, 10 and 26per cent. Two types of casein were used: a crude preparation possessing significant"extrinsic factor" activity and a purified casein with little activity.

2. The hematologic manifestations observed were (a) severe macrocytic anemia,(b) leukopenia, due to a proportionately greater reduction in polymorphonuclearthan in mononuclear cells, (3) slight thrombocytopenia, and (4) hyperplastic bonemarrow with an increase in immature nucleated red cells which resemble themegaloblasts seen in the bone marrow of patients with pernicious anemia.

3. The feeding of a 26 per cent rather than a 10 per cent crude casein diet did notprevent but did delay the onset of the blood changes. Anemia developed mostrapidly in the animals receiving 10 per cent purified casein.

4. The group receiving 26 per cent casein developed a greater degree of macrocytosis in the same period of time than did the group receiving 10 per cent casein.In all groups the degree of macrocytosis increased as the duration of the anemiaincreased.

5. The hematologic manifestations were not delayed nor was their developmentprevented by the intramuscular administration of 15 U.S.P. units of liver extractevery 15 days.

6. The blood and bone marrow returned rapidly to normal following the administration of pteroylglutamic acid, pteroyldiglutamic acid, pteroyltriglutamicand pteroylheptaglutamic acid. Thymine and xanthopterin had little or no activity. Tyrosine, adenine and uracil were inactive.

7. Purified liver extracts and crystalline vitamin B₁₂ were found to possess somehemopoietic activity in several animals but the activity was considerably less thanthat of the pteroylglutamic acid compounds.

8. The urinary excretion of "tyrosyl" (hydroxphenyl compounds) was notabnormal in the pteroylglutamic acid deficient pigs and was not altered by eitherpteroylglutamic acid or liver extract therapy.

9. The urinary excretion of allantoin and uric acid did not differ significantlyfrom the normal. Immediately following therapy with pteroylglutamic acid,however, in association with the reticulocytosis and lasting for the same period,there was a marked increase in the excretion of allantoin.

10. The results suggest that both pteroylglutamic acid and a factor in liverextract similar to or identical with vitamin B₁₂ are required for normal hemopoiesisin the pig.

Note: ACKNOWLEDGEMENTSThe crude methylfolic acid antagonist, xanthopterin, and the pteroylglutamic acid compounds, withthe exception of pteroylheptaglutamic acid, were kindly furnished by the Lederle Laboratories, PearlRiver, New York, through the courtesy of Dr. T. H. Jukes and Dr. S. M. Hardy.Sulfasuxidine was generously furnished by Sharp & Dohme, Inc., Philadelphia, Pa., through thecourtesy of Dr. W. A. Feirer.Pteroylheptaglutamic acid and Natola were supplied by Parke, Davis & Company, Detroit, Mich.,through the courtesy of Dr. A. E. Sharp and Dr. J. J. Pfiffner.Biotin was obtained from Hoffmann-LaRoche, Inc., Nutley, N. J., through the courtesy of Dr. E. L.Sevringhaus.The vitamins, with the exception of pteroylglutamic acid and biotin and including vitamin B₁₂ werekindly furnished by Merck and Company, Inc., Rahway, N. J., through the courtesy of Dr. A. Gibsonand the late Dr. D. F. Robertson.Experimental liver extracts (No. 1124, 1063, 1066 and 1067) were generously furnished by Armour andCompany, Chicago, Illinois through the courtesy of Dr. E. E. Hays.We are indebted to Mrs. Darlene Kehl, Mr. George Trappett, and Mr. Ocie Hadley for technicalassistance.

     

Experimental anemias in the rat; I. Macrocytic anemia in chronic pteroylglutamic acid deficiency and after splenectomy in Bartonella muris infection

1. In agreement with findings by other workers, rats in acute pteroylglutamicacid deficiency showed leukopenia and growth depression followed by death, without any significant change in the red cell picture.

2. In chronic deficiency, however, produced by the addition of small pteroylglutamic acid doses given intermittently, a severe anemia was obtained afterseventy days.

3. The anemia was macrocytic and "normochromic." Price-Jones curves showeda preponderance of macrocytes with anisocytosis. This agreed with findings byother workers for other species.

4. The anemia could be cured by single doses of 40 µg. or more of pteroylglutamicacid.

5. There was no significant difference between sexes to pteroylglutamic aciddeficiency. Reduction in the protein content of the diets, containing 1 per centsulfasuxidine, from 18 per cent to 10.5 per cent, produced no significant differencein the time of onset and severity of the blood symptoms.

6. These results were not due to infection with Bartonella muris. This infectionproduced a macrocytic anemia of a different type, and was curable by treatmentwith neoarsphenamine.

Note: ACKNOWLEDGMENTSWe are grateful to Dr. T. H. Jukes of the Lederle Laboratories for generous supplies of aldehyde-freePGA; and to Dr. K. Folkers of Merck Laboratories for the biotin used in these experiments. We wish tothank Dr. W. Jacobson for his advice during the course of this investigation. Valuable technical helpwas provided by Mr. D. R. Ashby, Mr. S. G. Impey, Miss M. J. Kemp and Mr. P. W. Wilson, to whomthe authors are indebted.

     

Effects of soy protein hydrolysate on blood pressure and angiotensin-converting enzyme activity in rats with chronic renal failure

We investigated the effects of soy protein and soy protein hydrolysate on blood pressure control, angiotensin-converting enzyme (ACE) activity, and renal function in a rat chronic renal failure model. Rats that had undergone a 5/6 nephrectomy were separated into three groups and fed different experimental diets for 14 weeks. At the end of the study, rats that fed a diet containing soy protein as the protein source had better blood pressure control and renal function, as well as lower circulating ACE activity and renal tumor-necrosis factor-alpha (TNF-alpha) concentration than rats fed a casein protein diet. Soy protein hydrolysate was shown to be as effective as soy protein in preventing the elevation of blood pressure, the progression of renal failure, and decreases in kidney TNF-alpha level, plasma ACE activity, and insulin concentration. In conclusion, the beneficial effects of consuming soy protein on blood pressure and renal function may be mediated mostly by its pepsin-digested hydrolysate through its ACE inhibitory activity.     

In vitro effects of x-radiation on white blood cells and blood platelets

1) Alkaline phosphatase activity of leukocytes is enhanced by radiation with50,000 r. This disturbance accentuates the inherent aging process of white bloodcells and may be explained by changes in the cell envelope.

2) X-radiation diminishes the endogenous oxygen uptake of leukocyte-plateletsuspensions by approximately 20 per cent. This response to radiation is demonstrable at exposures of as little as 5,000 r. The decreasing effect is diminishedwhen substrates such as sodium succinate or -glycerophosphate are added,within a wide range of their concentration. With increasing substrate concentration the decrease due to radiation approaches that of the endogenous respirationand even exceeds it in some of the experiments.

3) In pure blood platelets a similar decreasing x-radiation effect occurs forendogenous respiration as well as succinic dehydrogenase activity; -glycerophosphate dehydrogenase activity, on the other hand, is enhanced.

4) The oxygen uptake in leukocyte-platelet suspensions due only to leukocytescan be calculated. While the percentage radiation decrease of pure leukocytes isunchanged for endogenous and succinate activity, the decrease for -glycerophosphate as substrate reaches considerably higher levels (68 per cent compared with8.2 per cent in leukocyte-platelet suspensions). Thus -glycerophosphate dehydrogenase activity seems to be most sensitive to x-radiation. It was shown inone of our previous studies that -glycerophosphate dehydrogenase is one of themost important respiratory enzymes in leukocytes.

5) The glycolytic system in leukocytes remains intact following exposure toradiation with 50,000 r.

Submitted on October 5, 1956 Accepted on March 27, 1957     

Relationship between magnesium and zinc levels of the diet and its protein utilization

Protein utilization of the diet was tested in relation to different levels of magnesium and zinc in rats. The experimental diets contained either a low (0.14 g Mg or 10 mg Zn/kg) or adequate (0.45 g Mg or 40 mg Zn/kg) level of Mg or Zn and two different quality protein sources: casein or wheat gluten. Net protein utilization and net protein radio indexes in case of casein were significantly lower for the diet containing a low level of Mg or Zn. For gluten diets, such differences were not observed. Digestibility of protein measured in rats fed a low Mg or Zn casein diet was the same as for the diets with an adequate content of these minerals. Rats fed low Mg or Zn casein diets showed a significantly lower plasma Mg or Zn and a lower liver DNA content in comparison to the rats on adequate Mg or Zn diets. The results indicate that the utilization of protein is affected by Mg and Zn content of the diet and that this relationship depends on the quality of protein.     

Low Calcium Diet Enhances Development of Hypertension in the Spontaneously Hypertensive Rat

Young (5 week old) spontaneously hypertensive rats (SHR) were fed diets modified with respect to their calcium content. Control rats were given a normal calcium diet (0.3 per cent). During the experiment the systolic blood pressure (SBP), heart rate and serum level of total and ionized calcium were measured. Three diets were studied :calcium-free diet (0 per cent, h₁), low calcium diet (0.03 per cent, h₂) and high calcium diet (1.2 per cent, H). The h₁ diet induced a transitory increase (at week 2), then a long-lasting decrease in SBP for 12 weeks. Heart rate and calcemia were significantly decreased. The h₂ diet enhanced the increase in SBP and lowered heart rate for 10 weeks. Level of total and ionized serum calcium were unchanged. The high calcium diet (1.2 per cent) attenuated the increase in SBP for 44 weeks and enhanced the heart rate for 16 weeks. The serum level of total calcium remained stable but that of ionized calcium increased significantly at week 7. These data clearly establish that, in young SHR, a low calcium diet enhances the development of genetic hypertension and confirm earlier works obtained with calcium enriched diets. Experimental and clinical data lead us to emphasize the importance of alimentary calcium in the hypertensive pathology.     

Diets containing salmon fillet delay development of high blood pressure and hyperfusion damage in kidneys in obese Zucker fa/fa rats

Hypertension is the leading risk factor for cardiovascular and chronic renal diseases, affecting more than 1 billion people. Fish intake is inversely correlated with the prevalence of hypertension in several, but not all, studies, and intake of fish oil and fish proteins has shown promising potential to delay development of high blood pressure in rats. The effects of baked and raw salmon fillet intake on blood pressure and renal function were investigated in obese Zucker fa/fa rats, which spontaneously develop hypertension with proteinuria and renal failure. Rats were fed diets containing baked or raw salmon fillet in an amount corresponding to 25% of total protein from salmon and 75% of protein from casein, or casein as the sole protein source (control group) for 4 weeks. Results show lower blood pressure and lower urine concentrations of albumin and cystatin C (relative to creatinine) in salmon diet groups when compared to control group. Morphological examinations revealed less prominent hyperfusion damage in podocytes from rats fed diets containing baked or raw salmon when compared to control rats. In conclusion, diets containing baked or raw salmon fillet delayed the development of hypertension and protected against podocyte damage in obese Zucker fa/fa rats.     

An imbalance in the methionine content of the maternal diet reduces postnatal growth in the rat

The pregnant rat fed a low-protein diet has become widely used as a model system in the study of the prenatal programming of adult metabolism and disease. When pregnant rats of the hooded Lister strain were fed semisynthetic diets containing 18% or 9% casein supplemented with 0.5% dl-methionine, there was significant postnatal mortality in the group fed the low-protein diet. In a second experiment, dams were fed diets containing 9% casein supplemented with varying concentrations of dl-methionine up to 0.4% (w/w) and compared with a group fed a diet containing 18% casein supplemented with 0.5% dl-methionine. At birth, the pups from dams fed the low-protein diets supplemented with 0.2% dl-methionine or greater were significantly smaller than those of the dams fed the diet containing 18% protein. By 25 weeks of age, the body weight of the offspring of dams fed the low-protein diet supplemented with 0.2% or 0.3% dl-methionine were approximately 10% lower than those in the control group of offspring from dams fed 18% protein supplemented with 0.5% dl-methionine. There were corresponding changes in the weights of the major organs. These data suggest that increasing the dl-methionine supplement in the low-protein diet retards the growth of the fetus and affects the mature adult body weight. In contrast to the findings of other studies that used different formulas of the low-protein diet, the glucose tolerance in the offspring was unaffected by the protein content of the maternal diet at all levels of dl-methionine supplementation. These results suggest that the changes in metabolism of the offspring result from interactions between protein, lipids, and carbohydrates in the maternal diet, rather than a consequence of postnatal growth retardation per se and highlight the importance of considering all components of the maternal diet in the programming mechanism.     

A New Technic for Separation of Human Leukocytes

A simple technic for separation of human leukocytes is described. It utilizesdifferential sedimentation following red cell and platelet agglutination inducedby Polybrene. In this technic, lymphocytes are partially removed and over50 per cent of polymorphonuclears are recovered. The leukocyte/red cell ratiois about 5, if blood samples with normal leukocyte counts are used. By useof blood samples containing higher white cell counts this ratio may be increased. The separated cells proved to be highly viable as tested by phasemicroscopy, supravital staining and phagocytosis. There was no significantchange in oxygen uptake of leukemic cells in the presence of a relatively largePolybrene concentration.

Accepted on October 30, 1961     

Effects of soy components on blood and liver lipids in rats fed high-cholesterol diets

AIM: To assess the effects of soy protein, isoflavone, and saponin on liver and blood lipid in rats that consumed high-cholesterol diets. METHODS: High-cholesterol diets (1%) with or without soy material were fed to 6-wk-old male Sprague-Dawley rats for 8 wk. Blood lipids, liver lipids, glutamic oxaloacetic transaminase (GOT), and glutamic pyruvic transaminase (GPT) levels were measured. The in vitro bile acid-binding ability of soy materials was analyzed. RESULTS: The results of in vitro studies showed that soy protein isolate had a significantly higher bile acid-binding ability (8.4±0.8%) than soy saponin (3.1±0.7%) and isoflavone (1.3±0.4%, P<0.05). On the other hand, at the end of the experimental period, rats that consumed soy protein diets had lower GOT and GPT levels than rats that consumed casein under high-cholesterol diets. Rats that consumed soy protein also had lower total cholesterol (TC) levels in the liver than those that consumed casein under high-cholesterol diets. Rats that consumed the soy protein diet containing both saponin and isoflavone had lower hepatic TC level than those that consumed the soy protein diet without isoflavone alone. The effect of different types of proteins on triglyceride was not significant. CONCLUSION: Consumption of soy provided benefits to control lipid levels under high-cholesterol dieting conditions in this rat model of hypercholesterolemia. The major component that reduced hepatic TC was not saponin, but possibly isoflavone.     

STUDIES ON THE PHYSIOLOGY OF THE WHITE BLOOD CELL: THE GLYCOGEN CONTENT OF LEUKOCYTES IN LEUKEMIA AND POLYCYTHEMIA

The technic of determining glycogen in isolated white blood cells was appliedto the study of the different types of leukemia and of polycythemia, in order toobtain information on the physiology of the white blood cell. From this study itis concluded that the granulated leukocyte is the only carrier of glycogen in wholeblood. The "reducing substances" in lymphocytes and blast cells are not consideredas true glycogen.

The glycogen content of wet white blood cells in the rabbit amounts to about1 per cent. In the human being a range of from 0.17 to 0.67 per cent was calculated.In disease higher percentages occur, in polycythemia up to 1.64 per cent and inglycogen storage disease up to 3.05 per cent.

The glycogen concentration of normal white blood cells is within the same rangeas that of the striated muscle.

Note: I acknowledge with gratitude my indebtedness to Dr. William Dameshek for giving me the opportunity of analyzing the blood of some of the patients studied. Miss M. H. Campbell, Miss H. A. Clark,and Miss L. M. Garofalo have aided in carrying out many of the blood counts.

     

Malic and Lactic Dehydrogenase Isozymes of Normal and Leukemic Leukocytes Separated on Glass Bead Columns

Five LDH and two MDH isozyme bands were obtained with acrylamidegel electrophoresis of leukocyte extracts. Normal lymphocytes showed a hightotal H-LDH (heart type) activity (67 per cent) with 25 per cent in LDH-1and only 3.5 per cent in LDH-5. Lymphocytes from chronic lymphatic leukemia(CLL) and lymphosarcoma leukemia (LSA-LL) had less LDH-1, and moreLDH-3 and LDH-4, than normal lymphocytes. The H-LDH fell to 60.5 per centin CLL and 56 per cent in LSA-LL. PMN leukocytes had low H-LDH activity(38.8 per cent) with 3.3 per cent in LDH-1 and 25.8 per cent in LDH-5. Inmyelogenous leukemia, myeloblasts had the most LDH-1 and H-LDH, whilemature PMN had the least. PMN leukocytes isolated from CLL, LSA-LL, andmyelogenous leukemia had LDH patterns like the normal. Monocytes fromacute monocytic leukemia were low in LDH-1 and LDH-5, but had a high totalenzyme content. They evidently were rich in LDH-2, 3, and 4.

Lymphocytes had less MDH-1 (60 per cent) than PMN leukocytes (78 percent). In CLL, lymphocyte MDH-2 increased. In myelogenous leukemia,myeloblasts had the most MDH-2 and mature PMN the least. Monocytesfrom monocytic leukemia contained a little more MDH-2 than PMN leukocytes.

In general, white cell immaturity and/or ability to divide was associatedwith high levels of LDH-1, total H-LDH, and MDH-2.

Submitted on May 5, 1966 Accepted on June 26, 1966     

Effect of feeding maize/legume mixtures on biochemical indices in rats

Protein-energy malnutrition is one of the major public health problems in developing countries of the world due to prevailing socio-economic problems. This study aimed to observe the effect of formulated complementary blends on biochemical parameters of rats. Extruded complementary blends from maize fortified with cowpea or soybean at a level of 35% and 25% respectively were fed to 4 groups of rats for 28 days. Similarly, 3 other groups of rats were placed on casein, non-protein or rat pellet diet. Biochemical analysis was done on blood samples of the rats. Results from previous studies show the protein content of the formulated diets to range from 15.75% in UMC to 17.24% in MMS. Significantly (p < 0.05) lower WBC, Hb, MCHC, total protein, albumin and globulin values were recorded for the rats fed a non-protein diet (NP). The serum AST level was 75.5, 71.2, 63.2, 51.0, 60.5 and 55.7, respectively, for rats on casein, rat pellet, MMS, UMS, MMC and UMC (list of abbreviations is shown in the appendix) diets. Alkaline phosphatase was significantly (p < 0.05) higher in soybean-based diets while cholesterol was lowest in rats fed the non-protein diet (NP). The value obtained for serum electrolyte concentration in the rats fed NP compared well with rats on other diets but, however, had a significantly (p < 0.05) higher serum sodium value. These results confirm that the experimental diets supported growth, as shown in a previous study, and had no harmful consequence.     

Leukokinetic studies. I. A method for labeling leukocytes with diisopropyl-fluorophosphate (DFP32)

1. A method has been presented by which granulocytes can be labeledin vivo with diisopropylfluorophosphate containing radioactive phosphorus.The leukocytes are isolated from blood by dextran sedimentation of erythrocytes and are then treated with gramicidin and lysolecithin to remove remaining red cells. Platelets are removed by differential centrifugation. Theisolated leukocytes are placed between two squares of scintillating plasticand counted with a scintillation counter.

2. Leukocytes essentially free of erythrocytes and platelets can be obtainedby the method outlined. The efficiency of the plastic scintillation countingmethod for radioactive phosphorus is about 74 per cent and leukocyte samples obtained from 20 ml. samples of normal blood can be counted with areproducibility of ±10 per cent.

3. The administration of 2 mg. of diisopropylfluorophosphate either intramuscularly or intravenously is without significant toxic side effects.

4. No evidence has been obtained that the label damages the leukocytes.

5. No evidence has been obtained that the label elutes from leukocytesunder the conditions of these studies.

6. Diisopropylfluorophosphate labels granulocytes for a brief period oftime following injection. The label is not reutilized after death of the cells.

Submitted on June 16, 1958 Accepted on July 17, 1958     

The Recovery of Lethally Irradiated Dogs Given Infusions of Autologous Leukocytes Preserved at -80 C

Nine normal mongrel dogs were exposed to 1200 r whole-body irradiationat 4 to 5 r per minute. They were then given intravenous infusions of 2.12 to20 x 10⁹ autologous leukocytes that had been previously stored at -80 C. in10 per cent dimethyl-sulfoxide.

Three dogs survived with delayed but complete hematopoietic recovery.Three showed beginning marrow regeneration but died within 3-4 weeks ofirradiation. Three given less than 6 x 10⁹ cells died within 21 days. The numberof leukocytes infused was critical since there was no survivor among the dogsreceiving less than 9 x 10⁹ cells.

It is concluded that peripheral blood contains primitive cells capable ofrepopulating marrow spaces and restoring marrow function.

Submitted on May 8, 1963 Accepted on June 27, 1963     

A histochemical procedure for localizing and evaluating leukocyte alkaline phosphatase activity in smears of blood and marrow.

A rapid and simple method is described for demonstrating the presence ofalkaline phosphatase activity in cells of blood and bone marrow. Smears werefixed with 10 per cent formalin in methanol at 0 ± 5 C. for 30 seconds andstained by the azo dye procedure, using propanediol buffer at a pH of 9.6.

A "scoring" technic is outlined, which permits the comparison of alkalinephosphatase activities of blood leukocytes of different individuals.

Alkaline phosphatase activity was found only in neutrophilic granulocytesand was localized exclusively in the cytoplasm of these cells. In normal adults,43 to 95 per cent (mean of 78%) of all neutrophils were unstained. Staining wasup to ten times more intense in smears from patients with a variety of pathologic disorders.

Submitted on February 28, 1955 Accepted on June 25, 1955     

Hepatic and serum arginase and ornithine transcarbamylase activities of rats maintained on diets of different protein quality

Groups of rats were fed diets containing 15% protein from arhar dal (Cajanus cajan), bengal gram dal (Cicer arietinum), urad dal (Phaseolus mungo) and also isolated proteins from casein, egg, soya bean, gluten and gelatin for a period of 10 days. Rats maintained on the casein diet had the highest liver arginase activity and those having egg the lowest. All the leguminous proteins gave similar levels of arginase activity, the values falling between the casein and egg groups. Serum arginase level was found to be low only in rats on egg and gluten diets. Liver ornithine transcarbamylase (OTC) activity was significantly higher when legumes provided the protein as compared with the diet containing egg. Serum OTC level was lowest in the egg and arhar dal groups. Though the levels of urea cycle enzymes were altered by the quality of dietary proteins, no simple relationship could be established between them.     

Serum lipids, lipoprotein composition and liver cholesterol in genetically obese Zucker rats fed semipurified diets containing either casein or soy protein

The effect of semipurified diets containing either casein or soy protein on serum lipids, lipoprotein composition and liver cholesterol was studied in genetically obese Zucker rats. The ingestion of a cholesterol-enriched semipurified diet containing casein resulted in elevated levels of serum cholesterol and phospholipids compared to the feeding of a soy protein diet. No differences in serum triglycerides were observed. Differences in serum cholesterol and phospholipids were mainly reflected in the very low density lipoproteins and low density lipoproteins and to a minor extent in the high density lipoproteins. Liver cholesterol paralleled the levels of cholesterol in the serum, the rats fed casein exhibited markedly higher levels of liver cholesterol than those fed soy protein. Furthermore, the rats fed casein also had enlarged livers. Thus, this study clearly shows the differential cholesterolemic effect of dietary casein and soy protein in genetically obese Zucker rats.