Fast Hemoglobin in Lead Poisoning

1. An electrophoretically fast hemoglobin was found in approximately 40per cent of preschool children with elevated blood lead levels.

2. Fast hemoglobin was found more often in lead-poisoned patients withhypochromic anemia than in patients with normochromic red cells.

3. Fast hemoglobin differed from hemoglobins produced in vitro by incubation with chromate or oxidized glutathione. It had electrophoretic propertiessimilar to that found in a few patients receiving tolbutamide.

4. Fast hemoglobin could not be differentiated from normal hemoglobin A₃by any technic utilized.

5. Both lead and A₃ hemoglobins were heterogeneous molecular species.

6. The mechanisms leading to the production of hemoglobin A₃ and leadhemoglobin remain unknown.

Submitted on October 7, 1965 Accepted on December 10, 1965     

Lymphatic Drainage of Thymocytes to the Circulation in the Guinea Pig

1. An extensive network of lymphatic vessels coming from the thymus ofthe guinea pig and carrying thymocytes to the general circulation has beendemonstrated.

2. Local accumulations or infiltrations of cells at the surface of the lobuleor in the interlobular septa have been observed through which thymocytesappear to leave the thymus.

3. Irregular, tortuous canaliculi, resembling a lymph sinus in a lymph node,seem to play an important role in transporting thymocytes from these accumulations to the typical lymphatic vessels.

4. The output of thymocytes via lymphatic vessels to the general circulationis about 12.1 x 10⁶ per day.

Submitted on May 19, 1965 Accepted on September 11, 1965     

Quantitative Characterization of Chick Thymus and Bursa Development

1. Critical size analysis of the lymphoid cell populations of the developingchick bursa of Fabricius and thymus indicates that characteristic shifts in cellsize distribution, modal cell volume and mean cell volume occur during morphogenesis of these organs.

2. Comparison of bursa and thymus shows that while there are clear differences between the lymphoid cells of these two structures, the pattern of morphogenetic change is similar.

3. 19-nortestosterone injected in low doses can partially inhibit bursal development without affecting thymus morphogenesis. Depending on the age of theembryo at time of injection, however, doses sufficient to produce completebursal inhibition may also retard thymic morphogenesis.

4. The size differences found between bursal and thymic lymphocytes in thechicken parallel the size differences found between lymph node and thymiclymphocytes in the mouse.

Submitted on April 21, 1965 Accepted on July 10, 1965     

Brief Report: In Vitro Effect of Erythropoietin on the Spleen of the Polycythemic Mouse. I

1. Spleen explants of transfustion-induced polycythemic mice were incubated in vitro with and without erythropoietin.

2. After 24 hours incubation with the erythropoietin, immature large erythroblasts appeared, whereas the mature small erythroblasts were first observedafter 48 hours incubation.

3. Marked radioiron incorporation into the heme was observed after 48hours incubation with erythropoietin.

4. The control incubations without erythropoietin did not show these findings.

5. These data strongly suggest that erythropoietin induced the differentiation of erythroblasts from stem cells in vitro.

Submitted on July 19, 1965 Accepted on November 24, 1965     

Jordans' Anomaly in White Blood Cells: Report of Case

Persistent vacuoles were seen in the protoplasm of granulocytes, monocytesand occasional lymphocytes of 2 sisters suffering from ichthyosis. In the bonemarrow these vacuoles were found in the cytoplasm of promyelocytes, myelocytes, metamyelocytes and rarely in plasma cells. They were not observed inblasts or in red cells and thrombocytes and their precursors.

With the help of cytochemical staining technics and fluorescence microscopystudies, it was determined that the vacuoles contained lipids. A similar abnormality of leukocytes was described previously by Jordans in 2 brothers suffering from muscular dystrophy.

Ichthyosis was transmitted in this family in an autosomal recessive way.

Submitted on September 14, 1965 Accepted on December 10, 1965     

"Intermediate" cell types and mixed cell proliferation in multiple myeloma: electron microscopic observations

Bone marrow studies of multiple myeloma revealed, in some cases, aconspicuous proliferation of "lymphoid" cells, virtually indistinguishable bylight microscopy from those seen in lympho-proliferative disorders.

Electron microscopy demonstrated a variety of cells ranging from typicallymphocytes to cells with plasmocytoid features. Between these two types ofelements there were cells with intermediate characteristics.

In addition, in several cases of myeloma the presence of fixed reticuloendothelial cells and "reticular" plasma cells (or reticulum cells with plasmocyticfeatures) was frequently noted.

The presence of reticulum cells and lymphocytes and of cells apparently"intermediate" between these cellular elements and plasma cells, as judgedfrom electron microscopic observations, is suggestive morphologic evidenceof a phenomenon of cell transformation and evidence of a mixed cell proliferation in certain cases of multiple myeloma.

Submitted on February 16, 1965 Accepted on July 10, 1965     

Hydroxocobalamin: IV. Biological Half-Life of Hydroxocobalamin in the Human Liver

The mean hepatic biological half-life of Co⁵⁷-hydroxocobalamin injected to5 normal human subjects was similar to that of Co⁶⁰-cyanocobalamin, asshown by double-label hepatic surface counting during the first 30 weeks afterintramuscular or intravenous injection of cobalamins. In 2 cases in whomthe counting was extended over a year’s period, the clearance of hepaticradioactivity following the intramuscular injection of hydroxocobalamin hasslowed down as compared to that of cyanocobalamin, between the 30th and52nd week after injection.

Submitted on March 9, 1965 Accepted on May 18, 1965     

Effect of Anticoagulant and ABO Incompatibility on Recovery of Transfused Human Platelets

The effects of anticoagulant solutions on the recovery of transfused plateletswere studied. Citrate anticoagulants at pH 7.4 or pH 6.5 were found to beequally effective in preserving the viability of platelets when centrifugationof the cells was not required. When centrifugation is required, as in mostplatelet survival studies, citrate at pH 6.5 gives maximum recoveries. Ethylenediamine tetraacetate (EDTA) caused temporary sequestration of nearly alltransfused platelets and reduced maximum recoveries by about 50 per cent.

Platelet recovery was lowered by ABO-incompatibility between donor platelets and recipient serum, but survival time of remaining platelets was notaltered. Lowest platelet recoveries resulted when group A₁ or A₁ B plateletswere given to group O recipients with high isoagglutinin titers.

It is suggested that ABO-compatible platelets prepared in citrate should beused where possible in the treatment of thromboctyopenic disorders.

Submitted on January 5, 1965 Accepted on March 13, 1965     

The Influence of Chemotherapy on Survival in Acute Leukemia: III. A Comparison of Patients Treated During 1958-1964 with Those Treated in Two Sequentially Preceding Periods

Three-hundred and ninety-nine patients with acute leukemia examinedduring the period 1947-1964 were divided into three sequential series andtheir survival, from diagnosis to death, was compared.

A statistically significant increase in duration of survival for myeloblasticleukemia from a median of 2 to 5 months occurred between series I andseries III. This is the first convincing evidence that 6-mercaptopurine therapyinfluences survival in myeloblastic leukemia.

A steady increase in median survival of patients with lymphoblasticleukemia from 4 to 8 to 12 months was found in the three series. The resultsof analysis of these series are compared to other reported series. The hypothesis—that the longer survivals reported for patients treated in hematology clinicscompared to that of patients selected from population areas reflects patientselection—was examined and appeared unlikely.

Submitted on May 11, 1965 Accepted on August 6, 1965     

Antigenic Behavior of the S-63 Mouse Leukemia Virus

The present studies are an extension of earlier reports on the antibody response of mice to a leukemogenic virus (S-63). These investigations weredesigned to show whether a correlation exists between in vivo and in vitrodeterminations, and whether protective antibodies against the virus could beproduced in immune-tolerant animals. Viral neutralization and protectionstudies in mice were compared with passive cutaneous anaphylaxis (PCA),immunodiffusion, microprecipitin, and immunofluorescent tests using similarantigens and antibodies. As shown in tables 1, 2 and 3, antibody could bedemonstrated by both in vivo and in vitro methods, and these studies showed(table 2) that the antibody was protective.

The S-63 leukemia virus was isolated in 1963 from a mouse ascites cellleukemia line. That cell line had been carried in the laboratory for over 3years. The virus has been carried in ICR mice since its isolation. Immunologicstudies were carried out to determine the relation between the virus and theoriginating ascites cell. Studies of antibodies produced in normal rabbitsagainst the S-63 virus were unsatisfactory for purposes of these experimentsbecause of cross reaction with normal mouse tissue. Immune-tolerant animalshave yielded valuable information:

Antibodies produced in immune-tolerant rabbits injected with the S-63virus do not cross react against intact ascites cells from which the virus wasoriginally isolated. Conversely, antibodies similarly produced against theascites cells do not react against the virus antigen.

Antibodies produced in immune-tolerant rabbits against ascites cells protect mice against ascites cell-induced leukemia, but not against the diseaseinduced by S-63 virus. Mice challenged with ascites cells and treated withantiascites cell gamma globulin are protected against the disease. The besttherapeutic advantage is obtained when treatment is started either at the timeof cell inoculation or in the early stages of the disease.

Antibodies which protect newborn animals from infection develop in adultICR mice recovering from S-63 virus leukemia. These antibodies producedby convalescent mice are similar, from the standpoint of protection, to thoseproduced in immune-tolerant rabbits.

Submitted on January 14, 1965 Accepted on May 18, 1965     

Iron-deficient diet: effects in rats and humans

1. Iron absorption in rats is increased by dietary iron deprivation.

2. Erythropoiesis in the rat is unaffected by dietary iron deprivation thatincreases iron absorption by more than a factor of four.

3. Iron absorption is not increased in rats bled of an amount of iron equivalent to that lost in 5 days of iron deprivation.

4. These findings are compatible with the concept that iron-absorption iscontrolled by depletion of iron from a specific pool, separate from the hepaticand erythrocytic iron pools.

5. Iron absorption in human subjects was unaffected by dietary iron deprivation for 13 days. Reasons for differences between human and rat resultsare discussed.

Submitted on December 2, 1965 Accepted on March 26, 1966     

Extraction of an Erythropoietin-Producing Factor from a Particulate Fraction of Rat Kidney

1. A two-step method for the extraction of erythropoietin from hypoxic kidneys has been developed which allows residual plasma erythropoietin in renalvasculature to be separated from that of intracellular origin.

2. Renal extracts have been purified by DEAE cellulose chromatographyand found to contain 2 major erythropoietically active fractions. One bearsstrong resemblance to plasma erythropoietin. The other component is uniquein that it has practically no erythropoietic-stimulating activity unless previouslyincubated with normal rat serum. This activation phenomenon is used toidentify this kidney component as the renal erythropoietic factor (REF). TheREF has the capacity to produce erythropoietin or become erythropoieticallyactive when incubated with normal rat serum.

3. Differential centrifugation technics revealed that the REF is confined toparticles present in the light mitochondrial fraction of kidney.

4. Extracts of the light mitochondrial fraction of kidneys from normal ratsproduced significant amounts of erythropoietin when incubated with normalserum. The quantity found, however, was less than that evoked by similarextracts of kidneys from hypoxic rats.

5. The product of the incubation extracts of the renal light mitochondrialfraction with normal rat serum showed the same log dose/response regressionas sheep plasma erythropoietin standard.

6. It is hypothesized that either (a) the REF is a precursor of erythropoietinwhich must be complexed with a carrier present in normal serum in order tobecome physiologically active, or (b) the renal factor is an enzyme whichproduces erythropoietin by its action on a particular serum protein.

Submitted on October 1, 1965 Accepted on December 10, 1965     

Immunofluorescence of Megakaryocytes in the Thrombocytopenic Purpuras

The fluorescent antibody technic was applied to the study of thrombocytopenic purpuras with a presumable immunologic pathogenesis.

The results of the investigation suggest the hypothesis that some plasma protein material is strongly adherent to the surface of megakaryocytes in morethan 50 per cent of cases of chronic ITP.

Submitted on April 27, 1965 Accepted on October 25, 1965     

The Development of Hodgkin's Disease and Lymphoma during Anticonvulsant Therapy

1. Six patients in whom Hodgkin’s disease or lymphosarcoma developedduring the use of anticonvulsant agents (Dilantin, phenobarbital, Mysoline,Celontin) are reported.

2. The differential diagnosis from pseudolymphoma, the course of thesemalignant lymphomas, and their therapeutic management are discussed.

3. Based upon information presently available, it could not be determinedwhether the atypical hyperplasia sometimes observed during anticonvulsantdrug therapy is precancerous, or whether these agents may be carcinogenic incertain sensitive individuals.

4. Patients with a seizure disorder who develop lymphoma require standardtherapy for both conditions. The anticonvulsant drugs had no evident adverseeffect on the course of the lymphoma, although substitution of another agentseems warranted, if possible, for the offending drug.

Submitted on July 30, 1965 Accepted on January 31, 1966     

The Oxidation of Long-Chain Fatty Acids by the Formed Elements of Human Blood

1. In vitro, human erythrocytes free of contaminating white cells andplatelets do not oxidize fatty acid in the surrounding medium, either boundto albumin or supplied in buffer as the potassium salt. These results are notinfluenced by glucose deficiency.

2. White cells and platelets are both capable of oxidizing fatty acid fromthe surrounding medium—supplied either as the potassium salt in buffer orbound to albumin. Increasing the albumin/fatty acid molar ratio results in adecrease in the quantity of fatty acid oxidized. The amount of fatty acidoxidized is greater if glucose is not added to the medium.

Submitted on February 10, 1965 Accepted on May 31, 1965     

Inhibition of the Local Inflammatory Response in Man by Antimetabolites

The effects of chemotherapy on the local inflammatory response in man werestudied in 33 patients.

Inhibition of the mononuclear cell phase of the response at 4, 8 and 24hours was noted. The median on-therapy values at these hours were 0, 8.5and 23 per cent compared to control values of 4, 24.5 and 86 per cent, respectively.

The 24-hour values for the individual types of therapy were 13.5 per centfor intensive combination therapy with 4 antitumor agents, 21 per cent for6-mercaptopurine given i.v. daily for 5 days, 2 per cent for methotrexate giveni.v. daily for 5 days, 37 per cent for intermittent methotrexate and 56 percent for other therapy. Recovery was substantial within 5 days of the end oftherapy.

The inhibition was not correlated with alterations in the levels of circulatinggranulocytes or lymphocytes, nor with the numbers of granulocytes enteringthe inflammatory site.

Complicating infections were noted in eight patients, seven of whom werein the most strongly inhibited group. Observations of the response in ten untreated patients with acute leukemia showed both impaired granulocyte andmononuclear phases of the response.

Submitted on March 15, 1965 Accepted on May 18, 1965     

The Sites of Hemoglobin Catabolism

Hemoglobin is cleared from plasma principally by the liver, moderately bythe marrow, and relatively little by the spleen or other organs. Haptoglobinbinding has no effect on this relative distribution, its only discernible role inhemoglobin clearance or catabolism being that of preventing the renal lossof hemoglobin.

When bone marrow is depleted of parenchymal cells, as by cytotoxic chemicals or protein deprivation, its ability to take up hemoglobin from plasma ismarkedly enhanced.

Submitted on January 7, 1965 Accepted on April 23, 1965     

Human Bone Marrow Chromosomes in Megaloblastic Anemia

The long-recognized difference between chromosomes of megaloblastic andnormal human marrow cells can be measured in the cells studied by us. Thechromosomes from patients with megaloblastic erythropoiesis are approximately one-third longer than the corresponding normal chromosomes.

Submitted on January 18, 1965 Accepted on March 13, 1965     

Factors Controlling Erythropoiesis in Birds

1. Erythropoiesis in birds is stimulated by bleeding and environmentalhypoxia and is suppressed by induced polycythemia, indicating that the physiologic response is similar to that in mammals.

2. This response is mediated through a humoral factor which stimulateserythropoiesis of birds but not of mammals. Similarly, the erythropoietin ofmammals does not stimulate the erythropoiesis of birds.

3. Although both the erythropoiesis stimulating factor of birds and of mammals appear to require an intact protein structure for biological activity, theerythropoietin of birds differs from that of mammals in that it is not destroyedby sialidase and does not lose its activity when reacted with antibody to human urinary erythropoietin.

Submitted on June 7, 1965 Accepted on September 11, 1965     

The Effect of Erythropoietin on Hematopoietic Organs of the Polycythemic Mouse

Increased RNA-P³²/DNA-P³² ratio, iron incorporation and heme synthesisin spleens of polycythemic mice was demonstrated following an injection oferythropoietin. These changes were correlated with the appearance of erythroblasts. These findings suggest an intimate relation between RNA synthesisand stem cell differentiation.

Submitted on June 8, 1965 Accepted on July 25, 1965