Observations on the anemia of cryptogenetic splenomegaly. II. Expansion of the plasma volume

In cryptogenetic splenomegaly, although there is consistently a significantreduction in hemoglobin concentration and in the red cell count, the totalred cell volume may be reduced, normal or increased.

Following splenectomy the total red cell volume is either within normallimits or is increased, yet significant anemia persists.

Before splenectomy a significant increase in plasma volume is consistentlypresent. Following splenectomy the plasma volume falls, but it remains significantly greater than normal. It would appear that the greater the plasmavolume before splenectomy the greater is the fall following splenectomy.This fall in plasma volume is not a prompt response to splenectomy, for thefall which occurs one month after splenectomy is equalled or exceeded bythat occurring in the succeeding four months.

It is concluded that an expanded plasma volume before splenectomy contributes with the hemolysis previously reported to the development of theanemia encountered in cryptogenetic splenomegaly. Expansion of the plasmavolume at least contributes to the anemia which persists following splenectomy.

Submitted on February 12, 1957 Accepted on December 25, 1957     

Clot firmness

1. The relative firmness of clots in different systems is not necessarily directlycorrelated with the speed of clot formation.

2. Thrombin will hasten the development of full clot firmness, but will notsignificantly alter the degree of clot firmness of citrated plasma specimens, although the firmness of "purified" fibrinogen preparations clotted by thrombin isincreased with increasing thrombin concentration.

3. Citrate and/or calcium concentration influence clot firmness.

4. Fibrinogen preparations develop greater firmness on clotting with thrombinthan do citrated plasma preparations.

5. A platelet factor influences clot firmness of recalcified plasma, but not ofthrombin-activated or coagulase-activated plasma.

6. Clot firmness of recalcified citrated plasma is reduced by storage. This reduction in firmness is more rapid if the specimen is deep frozen as compared to unfrozen aliquots at 6 C.

Submitted on March 18, 1957 Accepted on July 16, 1957     

Alkali resistant type of hemoglobin in women with molar pregnancy

In 4 patients with molar pregnancy the fetal Hb blood concentration was foundto be significantly higher than that found in control groups of normal early pregnancy and in imminent and incomplete abortions. Following the molar abortionthe fetal Hb decreased gradually and reached normal vlaues within three to fivemonths.

It is assumed that the increased production of fetal Hb in these patients is dueto the reactivation of fetal erythropoietic activity by either excessive amounts ofchorionic gonadotropin, stress due to a rapidly growing tumor or to some otherunknown factors.

Submitted on April 17, 1957 Accepted on July 5, 1957     

Multiple hemangiomata associated with thrombocytopenia: remarks on the pathogenesis of the thrombocytopenia in this syndrome

1. A 13-year-old girl with multiple hemangiomata and thrombocytopeniais described.

2. Complications of surgical treatment included severe bleeding, septicemiaand a fibrinolytic event.

3. Simultaneous platelet counts from the tumor and peripheral blood wereperformed and showed a significantly higher concentration of platelets inthe tumor vessels.

4. With the rise in the platelet count, a decreased platelet fragility as wellas a persistently abnormal prothrombin consumption could be demonstrated.

5. The possible role of the tumor in the pathogenesis of the thrombocytopenia is discussed.

Submitted on December 27, 1957 Accepted on July 14, 1958     

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

1. The effect of diets, varying in quantity or quality of protein, on white bloodcell regeneration was studied in leukopenic rats, the leukopenia having been induced by a protein-free diet.

2. Diets containing different amounts of casein (3, 6, 9 and 18 per cent, respectively), were fed ad libitum. At the 3 per cent level, a further decrease occurred ofwhite blood cells, whereas the other three diets initiated a regeneration of leukocytes, its degree being more or less in proportion to the casein content.

3. In experiments with diets containing 18 and 30 per cent of casein, the amountof protein eaten and not its level in diet was the decisive factor in the regenerationof leukocytes. The white blood cell regenerating effect of a diet containing anoptimal level of protein, may be neutralized when given in restricted amounts.

4. Diets containing nutritionally inferior proteins, fed at 9 per cent level, alsoimpaired normal regeneration of leukocytes. The white blood cell regenerationafforded by the proteins investigated was found to increase in the following order:maize, gelatin, wheat, casein, processed soya, peanut, meat, egg.

5. In white blood cell regeneration promoted by dietary protein, granulocyteswere found to react to a greater degree than lymphocytes and monocytes.

     

In vitro incorporation of C14-DL-leucine into normal and leukemic white cells

Normal white blood cells and cells from patients with leukemia are able toincorporate C¹⁴-DL-leucine into proteins in vitro. Cells from individuals withchronic myelogenous leukemia were able to incorporate significantly greateramounts of the amino acid, and they were able to maintain this incorporation fora longer period of time than the control cells.

Both types of cells had the same in vitro survival time of four days, followingstorage at 3 C.

The incorporation was partially decreased in the presence of dinitrophenol, aswell as in an atmosphere of nitrogen.

Submitted on February 1, 1957 Accepted on March 7, 1957     

Selenoid (crescent) bodies

"Selenoid (Crescent) Bodies" are known in hematology as Cuerpos en MediaLuna in Spanish and Corps en demi-lune in French.

An original method of staining selenoid bodies and one which is suitablefor morphologic structural studies has been described. The method is alsosuitable for staining stromas of erythrocytes.

Selenoid bodies originate from erythrocytes and correspond to their stromas.Selenoid bodies can be produced in vitro by the spreading of blood on aslide. Two factors influence their formation: a mechanical factor by frictionof the erythrocytes against the surface of the slide and a chemical essentialfactor represented by the lipids of the blood, increasing the fragility of theerythrocytes.

The number of selenoid bodies in any smear of blood is directly proportional to its contents in lipids, either from an exogenous source (alimentarylipemia) or from an endogenous origin.

Selenoid bodies have also been found in the dog, sheep, rabbit, guineapig, rat and hen.

Submitted on August 8, 1957 Accepted on November 18, 1957     

Hematopoietic and nitrogen sparing effect of preparations from hog gastric mucosa with high content of blood group substances

A mucin fraction with high content of A and H blood group substances wasobtained from hog gastric mucosa. This fraction administered per os togetherwith vitamin B₁₂ in rats made anemic by phenylhydrazine injection showsa significant effect on the hematologic response in comparison with the animaltreated with vitamin B₁₂ alone.

The same fraction administered per os in cortisone-treated rats has alsoa significant nitrogen sparing action.

The possible mechanisms and relationships of the observed biologic effectsare briefly discussed.

Submitted on January 15, 1957 Accepted on September 23, 1957     

Polycythemia in primary carcinoma of the liver

In a series of 176 consecutive patients with hepatocarcinoma an increasein red cell count and in hemoglobin significantly above normal levels wereencountered in 17 (10 per cent).

An investigation of 28 patients with hepatocarcinoma developing in acirrhotic liver is reported. In three of the patients (10 per cent), the redcell counts and hemoglobin levels were significantly above those encounteredin healthy Chinese.

In these 28 patients it has been shown that the plasma volume is increased,and this increase does not differ significantly from that encountered in uncomplicated cirrhosis of the liver. The total red cell volume, on the otherhand, is significantly greater than in uncomplicated cirrhosis of the liver. Whilethe mean total red cell volume in hepatocarcinoma is not significantly different from that in healthy controls, consideration of this finding in individualpatients shows that it was above normal in 17, normal in six, and in the remaining five it was below normal.

It is concluded that the polycythemia encountered is a true polycythemiasecondary to the development of the hepatocarcinoma. The expanded plasmavolume is considered probably attributable to the pre-existing cirrhosis ofthe liver. Unfortunately, in the course of this investigation we did not encounter a patient in whom the carcinoma had developed in a liver whichwas not cirrhotic.

Submitted on April 8, 1957 Accepted on November 18, 1957     

An abnormal hemolytic system associated with leukemia and other disseminated malignant diseases

1. In the blood of some people with leukemia and other disseminated neoplastic diseases it is possible to demonstrate an abnormal hemolytic mechanismby incubating sterile blood in vitro at 37 C. for 24 hours. There is a latent periodof 6 to 8 hours before hemolysis begins. Hemolysis can be inhibited by an excessof glucose or by low pH and also by removal of calcium. The heat stability of thesystem indicates that the plasmal abnormality is not enzymatic.

2. It is suggested that hemolysis is due to a reaction between ionic calcium andsome component of the erythrocyte. The nature of the plasma abnormalitythat permits the reaction is not known.

3. The significance of this phenomenon in the pathogenesis of anemia in neoplastic disease has not been established.

Submitted on January 29, 1957 Accepted on March 27, 1957     

On the occurrence of a plasminogen-like substance in human tissues

Evidence is presented for the occurrence of a plasminogen-like proenzymein human tissues.

When homogenates of human connective tissue are incubated with SKthey dissolve fibrin clots far more easily than control homogenates withoutSK. This dissolution of fibrin clots by tissue extracts is a proteolytic reaction similar to the dissolution of fibrin clots by plasma enzyme.

The chemical features of tissue "plasminogen" are the same as of bloodplasminogen. It can be extracted at very high and very low PH, precipitatedat pH 5.3 (isoelectric point), and inhibited by ions of heavy metals in exactlythe same manner as blood plasminogen.

A similar behavior was found in the case of tissue plasminogen in accordance with recent views on the complex nature of the blood fibrinolyticsystem. The occurrence of tissue inhibitors of plasmin was proven, and itwas shown, on comparing the activity of human and bovine tissues, thatthere is the possibility that in tissues there also exist a "proactivator" and"plasminogen" in accordance with Müllertz and Astrup.

Tissue "plasminogen" occurs chiefly in organs rich in connective tissue,like the aorta wall, fascia, etc. In other organs the "plasminogen" activityis negligible.

It is suggested that tissue "plasminogen" may be of significance in thepathogenesis of certain diseases of connective tissue, e.g. rheumatoid arthritis, in which proteolysis is postulated to be an important pathogenicfactor.

Submitted on March 18, 1957 Accepted on October 10, 1957     

Starch block electrophoretic studies of human hemoglobin solutions. II. Results in cord blood,thalassemia and other hematologic disorders: comparison with Tiselius electrophoresis

The results of starch block electrophoresis of normal adult hemoglobinin the various disease states has been reported. The slow-moving fraction,A₂, is elevated in all patients so far studied with thalassemia. Some patientswith Addisonian pernicious anemia also show an increase in this fraction.

Cord blood contains very little A₂.

The A₂ fraction has no correlation with the severity of the disease.

The A₂ fraction found on starch seems to be identical with the small,fast-moving component noted in the Tiselius apparatus at acid pH.

Iron deficiency anemia of the acquired type tends to have a lower amountof A₂ than normal, with an increase to average normal values after therapy.

Submitted on September 9, 1957 Accepted on December 15, 1957     

Unique sequence of pernicious anemia,polycythemia, and acute leukemia

The development of three primary blood dyscrasias, namely, perniciousanemia, polycythemia vera, and acute myeloblastic leukemia, in one patient hasbeen reported.

Submitted on March 22, 1957 Accepted on July 1, 1957     

The effects of acute protein depletion on the distribution of radioiron in rat tissues

1. The distribution of Fe⁵⁹ in plasma, bone marrow, erythrocytes, liver, andspleen was determined at specified intervals after single intravenous injectionsof the radioisotope into normal and protein-deficient rats.

2. In normal rats, injected radioiron followed the usual metabolic pathways; i.e., through bone marrow to erythrocytes, with small, consistent amountsto liver and spleen.

3. In protein-deficient animals normal Fe⁵⁹ time-concentration relationshipswere disrupted completely.

4. Less iron was recovered from the tissues of protein-deficient rats thanfrom normal animals.

5. Storage iron in liver and spleen was not conjugated with polysaccharide,even though tissue protein was seriously depleted.

Submitted on June 27, 1957 Accepted on September 15, 1957     

The postnatal synthesis of fetal hemoglobin. I. Some studies in newborn goats using radioactive Fe59 and glycine-2-C14

Postnatal synthesis of fetal and adult hemoglobin was studied with radioactiveiron (Fe⁵⁹) and glycine-2-C¹⁴ in newborn goats.

At birth about 60 per cent of Hb-F is present in newborn goats; after 60 daysonly about 5 per cent is found. Two goats were treated with "exchange transfusions" with blood of the mother. Using an alkali denaturation method the fetalhemoglobin was not demonstrable during the first few days after this treatment,but 20 days later about 20 per cent was present in circulating blood.

Maxima of radioiron-specific activity in adult hemoglobin occurred in untreated newborn goats at the 12th day, of C¹⁴ at the sixth day; in "exchangegoats" for radioiron at the 8th day. The fetal hemoglobin maxima of specific activity occurred in the untreated goats resp. after 32 and 24 days, in "exchangegoats" after 8 days.

The reappearance of Hb-F in "exchange goats" and the specific activity ofHb-F are direct indications for a synthesis of this component after birth inthis species.

Submitted on January 29, 1957 Accepted on May 24, 1957     

Albumin synthesis in young and elderly subjects using a new stable isotope methodology: Response to level of protein intake

Albumin synthesis was evaluated in 5 young adult males (19–25 yr) and 6 elderly males (64–78 yr) by a procedure involving oral administration of ¹⁵N-glycine every 3 hr over a 60-hr period. From about 40 hr onwards, urinary urea achieved a plateau of ¹⁵N-enrichment, which was estimated from the average of the last five (low protein) or seven (adequate protein) consecutive three-hourly urinary samples of the 60-hr period. This enrichment plateau was used as an index of the ¹⁵N-enrichment of the guanidine N of hepatic free arginine. The ¹⁵N-enrichment of the guanidine N of arginine in serum albumin was determined and albumin synthesis was estimated by comparing this value with the estimated enrichment of precursor hepatic arginine. Using this methodology, serum albumin concentration, synthesis, rate and plasma volume were measured when the young and elderly subjects had received an adequate protein intake (1.5 g · kg^?1 for 7 days) or a low protein intake (0.4 g · kg^?1 for 14 days). Serum albumin concentration was lower in the elderly at both levels of protein intake; protein intake did not affect this parameter in either age-group. Plasma volume (per kg body weight) did not differ between young and old, but increased in both groups when they were given the low-protein diet, so that the total intravascular albumin mass increased in both age groups significantly in the case of the young, and was probably due to net transfer of albumin from the extravascular pool. The fractional synthesis rate of the whole body albumin pool with adequate intake of protein was 4.0%/day in the young and 3.4%/day in the elderly. This fractional rate was reduced significantly by giving the low-protein diet to the young subjects, but was not reduced in the elderly. Absolute synthesis rates, calculated per kg body weight and per kg body cell mass, led to a similar conclusion. Whole body protein synthesis was also estimated from urinary ¹⁵N-urea enrichment using the Picou and Taylor-Roberts model. Albumin synthesis as a percentage of whole body protein synthesis (5%–6%) was reduced in the young adults by giving the low-protein diet, but was unchanged in the elderly. In conclusion, the rate of albumin synthesis in the young, but not in the elderly, is sensitive to changes in protein intake. It is suggested that albumin synthesis in the elderly is controlled at a lower set point, which prevents its response to higher protein intakes.     

The effects of splenectomy on the red blood cells of the dog with particular emphasis on the reticulocyte response

In splenectomized dogs the following observations on the erythroid elementswere made:

1. There was no unusual postoperative depression of erythrocyte, hemoglobin or hematocrit values, and regeneration occurred at the same rate as inthe control dogs.

2. A significant increase in the number of circulating reticulocytes wasnoted, which persisted for a longer duration than the associated thrombocytosis. There is a suggestion that the reticulocytes circulating during regeneration were more immature than those of the control animals.

3. Varying numbers of target cells accompanied by increased osmotic resistance were present in all the splenectomized dogs. Most also developednormoblastemia, particularly during active regeneration, which persistedthereafter in some of the animals. Howell-Jolly bodies were rare.

4. No significant bone marrow changes were noted.

It is suggested that:

1. In nonsplenectomized animals, most erythrocytes released from the bonemarrow in response to blood loss are mature red cells rather than reticulocytes.

2. Postsplenectomy reticulocytosis is not dependent on a fall in hematocritand is not a reflection of hemolysis or even of increased erythropoiesis, except,perhaps, in part during regeneration from the blood loss incurred at surgery.

3. Postsplenectomy reticulocytosis and normoblastemia are unaccompaniedby alterations in the myeloid-erythroid ratio of the bone marrow and areprobably largely due to a diminution of the normal inhibition of erythroid release from the marrow, permitting their earlier entry into the circulation.

4. The aforementioned changes are due specifically to the removal of thespleen itself, rather than merely to the removal of splenic blood from theportal circulation.

Submitted on November 18, 1957 Accepted on April 12, 1958     

Chediak''s anomaly of leukocytes in malignant lymphoma associated with leukemic manifestations: case report with necropsy

A case of Chediak’s anomaly of leukocytes was described, the fourth reported in the literature.

The patient was an 11-month-old boy who, in addition to Chediak’s anomalyof the leukocytes, presented a leukemic blood picture.

He died five days after admission to the hospital, and necropsy revealeda malignant lymphoma.

We consider it possible that in all the reported cases there were generalizedpathologic changes in the reticulohistiocytic system.

Submitted on December 3, 1957 Accepted on May 15, 1958     

The nature of the hemorrhagic disorder accompanying hemolytic transfusion reactions in man

A hemorrhagic diathesis has been observed in 2 patients who received 500 ml.of incompatible whole blood. In both, hypofibrinogenemia, hypoprothrombinemiaand thrombocytopenia were observed and there was no evidence of increasedfibrinolytic activity. In one, accelerin activity was reduced and there was transient evidence of a low-titered circulating anticoagulant.

The most likely explanation for the observed changes is intravascular coagulation in the recipient, presumably initiated by the thromboplastin-like activityof the hemolyzed red blood cells.

Submitted on November 5, 1956 Accepted on January 11, 1957     

The efficiency of oxidative phosphorylation by normal and leukemic human leukocytes

1. A new modification of existing methods has been described for the separation of leukocytes from whole blood which provides a procedure for therapid isolation of uninjured cells suitable for the study of oxidative phosphorylations.

2. This method has been employed in a study of the relative efficiencyand yield of oxidative-linked phosphorylations mediated by normal and leukemic or immature leukocytes. The maximum aerobic phosphorylating capacitywas exhibited by chronic lymphocytic leukemic leukocytes, followed in decreasing order of activity by acute monocytic leukemic leukocytes and chronicmyelocytic leukemic leukocytes. Oxidative phosphorylation was not demonstrated with normal leukocytes.

3. Results of this study suggest that expression of leukocyte metabolicdata on a unit nitrogen basis more accurately reflect the morphologically obvious size differences among the various leukocytes than presentation of dataon a unit cell basis.

4. The aerobic phosphorylations mediated by leukemic leukocytes werefound to be dependent upon substrate addition and were depressed by lowlevels of dinitrophenol. Under the experimental conditions employed in thisstudy, glucose-6-phosphate was formed in stoicheiometric amounts. These results indicate that leukemic leukocytes are capable of the aerobic esterification of inorganic phosphate accompanying the oxidation of selected Kreb'scycle intermediates.

Submitted on August 2, 1957 Accepted on November 2, 1957