In order to investigate the erythroid committed stem cells in polycythemia vera, the marrow cells from 8 patients in relapse and 2 patients in remission were cultured with erythropietin in vitro according to the method of Krantz. The marrow cells from all 8 patients in relapse showed low or no response to erythropoietin, comparing with that of normal volunteers. The marrow cells from 2 patients in remission responded to erythropoietin to the almost same degree as that of normal volunteers. Considering the action of erythropoietin, which mainly acts on erythroid committed stem cells, inducing their differentiation into erythroblasts, there might be some abnormalities in erythroid committed stem cells in polycythemia vera. Moreover, it might be possible that normal erythroid committed stem cells also exist in the marrow cells.
The colony formation in soft agar of the marrow cells from polycythemia vera patients was investigated by the method of Robinson. The number of colonies formed in soft agar which is considered to reflect the granuloid committed stem cells, was increased in relapse and was almost same as that of normal volunteers in remission. From these findings, it could be considered that the granuloid committed stem cells is increased in polycythemia vera in relapse.
These findings were discussed in relation to the pluripotent stem cells and the pathogenesis of polycythemia vera.

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Data of 217 patients with polycythemia vera diagnosed since 1960 were collected in Japan. The mean age of diagnosis was 53.0 years and a male: female ratio was 1.1:1.
Forty six patients were treated with P³² and 133 patients were treated by chemotherapy. In the course of chemotherapy busulfan was used most frequently.
The survival rate of the series was 75 per cent at 5 years and 55 per cent at 10 years after diagnosis. The survival was shorter in older patients and in those who had hypertension. After an adjustment for the age of patients was made, there was no difference between the survival of patients treated with P³² and that of patients treated by chemotherapy.
Of 31 patients who had died, the cause of death were vascular lesions in 8, anemia (myelofibrosis or aplastic anemia) in 4, acute leukemia in 2, cancer in 2, and others.

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1) Twenty-two cases bearing various tumors associated with erythrocytosis have been reported in Japan. Through analysis of 457 cases with various tumors at the Keio University Hospital, the incidence of erythrocytosis is estimated 2.2∼4.4% in renal carcinoma; 1.9∼3.8% in hepatoma; lower than 1% in uterine myoma, various brain tumors and gastric carcinoma.
2) The first case of a 38-year-old male having meningioma associated with erythrocytosis was reported. Erythropoietin (Ep) activity estimated by polycythemic mouse bioassay in the pre-operative serum and the tumor extract was elevated, suggesting that this tumor was “Ep producing tumor”. The estimation of Ep in 3 cases with gastric carcinoma possibly associated with erythrocytosis failed to suggest that gastric carcinoma is a variety of “Ep producing tumor”.
3) In renal adenocarcinoma associated with erythrocytosis, Ep activity in the tumor extract and pre-operative serum was elevated and Ep inhibitor normally present in the kidney tissue disappeared in the tumor extract.
4) In uterine myoma associated with erythrocytosis, Ep activity was elevated in the pre-operative serum and tumor extract. The elevated activity in the latter, however, was different in quality from that of the renal carcinoma. Ep inhibitor seemed to be not important for production of erythrocytosis.
In conclusion, careful examination of the patients bearing various tumors may lead one to find more cases of “Ep producing tumor”. The Ep estimation in the tumor extract is the most important for the diagnosis of “Ep producing tumor”.
It is suggested that there are at least two different mechanisms in the production of erythrocytosis associated with various tumors.

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Fifteen abnormal hemoglobins which cause or may cause erythrocytosis are reviewed. Their clinical expression is a benign hereditary polycythemia which affects only erythrocytes. Abnormally high oxygen affinity of hemoglobin causes diminished oxygen supply to the tissue, and this in turn stimulates erythropoietin production by the kidneys resulting in increased erythropoiesis in the bone marrow. Hb Hiroshima is discussed first as a representative case, and the functional characteristics of such abnormal hemoglobins are explained in relation to their structural abnormality or a single amino acid substitution. Importance of blood oxygen affinity as a screening test for abnormal hemoglobin in erythrocytosis or anemia is stressed.

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From erythropoiesis-stimulating activity of various hormones, relationships between hormones and erythrocytosis have been investigated, clinically and experimentally.
The results obtained were as follows:
1. Out of five cases of patients with Cushing's syndrome resulting from overproduction of glucocorticoid, only a case associated with erythrocytosis was observed. It was thought as a cause of erythrocytosis that glucocorticoid increased oxygen consumption in tissue.
2. Erythrocytosis was observed in a patient with vilirizing adrenal carcinoma, and in two cases out of twenty patients with breast cancer under the treatment with large dose of testosterone also. Mechanisms by which androgen stimulates erythropoiesis may be related to a direct stimulatory effect on hemopoietic stem cell and to potentiate activity of erythropoietin.
3. Out of nineteen cases of patients with hyperthyroidism, eleven cases were associated with hypochromic anemia and it was pointed out that a main cause of anemia in such patients was likely to be iron deficiency. From experimental results it was suggested that thyroxin and L-triiodothyronine have a stimulatory effect on erythropoiesis indirectly through their calorigenicity.
4. None of patients with acromegaly was associated with erythrocytosis. However, from experimental results it was suggested that growth hormone itself shows an erythropoietic action possibly due to stimulation of erythropoietin production without an increasing oxygen consumption, differing from mechanisms of both thyroxin and glucocorticoid.

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Marked erythrocytosis has been determined for two of the four siblings born to parents of a consanguinous marriage.
The elder brother (25) had had a ruddy complexion since infancy. Erythrocytosis was detected at the age of 19 when he was affected by a common cold. He had a slight case of splenomegaly. The younger brother entered the hospital, likewise at the age of 19, with a complaint of recurrent headaches since the age of 15. He had a slight case of splenomegaly and simple goiter, Marked erythrocytosis, rise in Ht, increase in Hb, and increase in total red cell volume were detected in both patients, Leukocytosis and thrombocytosis were not detected in either of them. The differential white cell counts were normal. There was hyperplasia in the marrow. Half life of red cells by ⁵¹Cr was within normal limits. Urine protein tests showed positive for both patients but no abnormalities were detected by the renal function tests and pyelography. Arterial oxygen saturation was normal. Abnormal hemoglobin could not be found. The oxygen affinity of Hb for the equilibrium curve (taken for the elder brother) was normal. Other diseases which may have led to secondary polycythemia could not be detected. Tests employing the polycythemic mouse assay showed marked increase in erythropoietin activity both in plasma and urine. ³²P treatment was given to the elder brother but this only resulted in a slight decrease in the red cell volume. At the age of 23, the elder brother was affected by celebral thrombosis but this was cured by phlebotomy. Vercyte and letting was effective in eliminating the headaches for the younger brother. They are leading a healthy normal life at the present.
Familial erythrocytosis is a disease characterized by increase in red blood cells without leukocytosis and thromboeytosis and by good prognosis. There have been only 24 cases reported in literature including the present two cases. It is suspected that there is autosomal recessive type of inheritance but this is not definite. With the exception of one case, increase in erythropoietin activity has been reported in 6 families. It is believed that this disease results from the overproduction of erythropoietin due to congenital aberrant regulatory mechanism for erythropoietin production. We have also studied the reports on polycythemia caused by abnormal hemoglobin and other polycythemic disorders in relation to the present disease and the problems involed in the classification of the disease therein.

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The glass bead column method for determining platelet adhesiveness is described. The method is a modification of the original technique of Salzman as follows.
A nylon tubing containing glass beads was connected with a first syringe using a three-way stopcock. Test blood sample (being filtered) as well as control blood sample was drawn using one needle, which was attached to the other side of the stopcock. The test blood sample was collected through glass bead column into a second syringe which contained 3.8% citrate and was distally connected with the nylon tubing. The filtration rate of test blood sample was 2.7 ml/30 seconds.
Platelet factor-3-activity as well as its availability and adenine nucleotide contents in filtered platelets was not significantly different from those in control platelets. Platelet aggregability was slightly increased after filtration through glass bead column.

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A urinary B₁₂ excretion test by dual isotope technique was performed in 22 patients including 7 with pernicious anemia, 3 with total gastrectomy and 2 with chronic renal failure. The urinary excretion rate of ⁵⁸Co-B₁₂ was 21.3±3.7% in control group and 4.1±2.6% in both patients with pernicious anemia and those with total gastrectomy. The ratio of ⁵⁷Co-B₁₂ to ⁵⁸Co-B₁₂ was significantly high (p<0.01), ranging 1.4 to 9.4 in the patients with IF deficiency. In this group, percent excretion of ⁵⁸Co-B₁₂ was greater than that of free B₁₂ only when administrated to the same patient.
Free IF was not found in the solution of IF bound ⁵⁷Co-B₁₂ and 8 to 15% of ⁵⁷Co-B₁₂ was proved free from IF by coated charcoal method. It was showed that the exchange of ⁵⁷Co-B₁₂ on IF with free B₁₂ occurred depending on pH and temperature in vitro. Therefore, the higher urinary excretion of the free form of B₁₂ in the dual isotopic method was probably due to the exchange of two forms of B₁₂ in vivo.
The radioactivity of ⁵⁸Co-B₁₂ in 2 ml of urine is too low to be measured accurately. This was settled by counting ⁵⁸Co-B₁₂ adsorbed on uncoated charcoal from 20 ml of urine.

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Case 1. A 57-year-old woman, IgA plasma cell leukemia without Bence Jones proteinuria.
In this case, the nuclei of the leukemic cells exhibited indentation and lobulation similar to that seen in Rieder cells in acute myeloid leukemia. Giant plasmacytoma cells were also seen in the bone marrow smear and section. Electronmicroscopically, almost all the myloma cells showed the presence of polyribosome clusters formed by 10∼15 ribosomes, and poor developement of mitochondria and rough-surfaced endoplasmic reticulum presenting short-tubular from.
The deformity of the nuclei and the appearance of giant cells seemed to resemble to those of the reticulum cell.
Case 2. A 59-year-old man, plasma cell leukemia with λ-typed Bence Jones proteinuria.
In the electronmicroscopic observation, some of the ergastoplasmic cisternae were with the leukemic cells showed poor developement of Golgi zone, and well developed lamellar structure and others with dilated vesicular form. The cytoplasma also contained small polyribosome clusters between the nucleus and the endoplasmic reticulum.
The correlation between the presence of small polyribosome and high L chain synthesis was discussed.

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A case of β-thalassemia minor was found in Fukuoka city recently. The proposita, 52-years-old housewife with no symptoms, admitted on December in 1972, for purpose of the examination of anemia which was detected one year ago and refractory to the administration of iron, vitamin B₆ and viamin B₁₂.
Laboratory examination revealed microcytic, hypochromic anemia associated with morphological abnormality of erythrocytes, normal serum iron level, erythroid hyperplasia of bone marrow, increased osmotic resistance and shortened life span of erythrocytes. Homoglobin A₂ and hemoglobin F were increased significantly in comparison with normal value.
Her husband, son and three daughters were examined. Her son and two daughters except the eldest one had microcytic, hypochromic anemia associated with the same hematologicl findings as proposita had. They also had increased hemoglobin A₂ and hemoglobin F.

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