The Lipid Distribution of Human Platelets in Health and Disease 1

Erickson, Betty Nims; Williams, Harold H.; Avrin, Ira; Lee, Pearl

doi:10.1172/jci101029

Cited by 23 publications

(9 citation statements)

References 14 publications

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“…These workers also reported at that time what has recently been emphasized again (7,8): phosphatides from other natural sources (soy bean, yeast) also were capable of accelerating blood clotting in vitro. Erickson, Williams, Avrin and Lee (9), confirmed the results of Chargaff and associates relating to platelet phospholipid content. Wallach, Surgenor and Steele (10), identified phosphatidyl ethanolamine and phosphatidyl serine in human platelets.…”

supporting

confidence: 78%

Thromboplastic Factors in Platelets and Red Blood Cells: Observations on Their Chemical Nature and Function in in Vitro Coagulation *

Troup¹,

Reed²,

Marinetti³

et al. 1960

J. Clin. Invest.

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Although the existence of blood platelets had been known for many years it was not until 1882 that Bizzozero (1) called attention to the role of platelets in blood coagulation. Morawitz (2), twenty years later, was among the first to consider the specific chemical contribution of the platelet to the clotting process.Howell (3), in 1912, demonstrated the presence of an unsaturated phosphatide ("kephalin") in thromboplastic substances of tissue origin. Mills (4), in 1927, showed cephalin to be present in blood platelets, and the following year Haurowitz and Sladek (5) published results of chemical analysis of horse platelets, reporting a lipid content of 12 per cent of dry weight. The first convincing data relating the cephalin content of platelets to acceleration of coagulation were supplied by Chargaff, Bancroft and Stanley-Brown (6). These workers also reported at that time what has recently been emphasized again (7,8): phosphatides from other natural sources (soy bean, yeast) also were capable of accelerating blood clotting in vitro. Erickson, Williams, Avrin and Lee (9), confirmed the results of Chargaff and associates relating to platelet phospholipid content. Wallach, Surgenor and Steele (10), identified phosphatidyl ethanolamine and phosphatidyl serine in human platelets. Subsequently the presence of inositol phosphatide was noted ( 11,12 A. Isolation of platelets from whole blood. Blood was obtained from normal adult volunteers; it was collected by gravity flow into plastic bags containing 1.5 per cent disodium dihydrogen ethylenediaminetetraacetate in 0.7 per cent NaCl,1 employing a plastic donor set.2 A proportion of 9 volumes of blood to 1 volume of anticoagulant was used. Forty ml aliquots were then gently decanted into silicone-coated3 glass tubes and these were centrifuged at room temperature at 300 G for 15 minutes. All glassware used in collection and transfer of platelet-containing plasma was similarly treated. All glassware used in the coagulation studies was acidcleaned. The supernatant platelet-rich plasma was transferred by pipette to glass centrifuge tubes. Aliquots of platelet-rich plasma were then taken for platelet and white cell counting, and aliquots to be used in coagulation studies were stored at 4°C in glass tubes. The platelet-rich plasma thus obtained had fewer than 300 leukocytes per cu mm, and was completely free of red cells. The platelet-rich plasma was then centrifuged at 2,000 G at 40 C. The supernatant platelet-poor plasma thus obtained was decanted into other centrifuge tubes and again centrifuged at 40 C for 30 minutes at 2,000 G. This supernatant plasma had fewer than 1,000 platelets per cu mm (49) and was considered to be "plateletpoor." The sedimented platelets obtained after the first 2,000 G centrifugation were pooled and washed 3 times with isotonic NaCl solution and resedimented by centrifugation at 40 C for 5 minutes at 900 G. The platelets thus obtained were then subjected to lipid extraction and analysis.B. Extractiont, separation, and measurement of platelet and...

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supporting

confidence: 78%

Thromboplastic Factors in Platelets and Red Blood Cells: Observations on Their Chemical Nature and Function in in Vitro Coagulation *

Troup¹,

Reed²,

Marinetti³

et al. 1960

J. Clin. Invest.

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“…The phospholipid composition of human platelets and erythrocytes has been reported by several investigators (1)(2)(3)(4) and the metabolism of phospholipids in human blood has been studied in some detail (5)(6)(7)(8)(9). Recent investigations of phospholipid metabolism of leukocytes have been reported (6, 10) but we are not aware of similar studies on platelets.…”

mentioning

confidence: 94%

The Incorporation of Radioactive Phosphorus Into the Phospholipids of Human Leukemic Leukocytes and Platelets*

Firkin¹,

Williams²

1961

J. Clin. Invest.

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The phospholipid composition of human platelets and erythrocytes has been reported by several investigators (1-4) and the metabolism of phospholipids in human blood has been studied in some detail (5-9). Recent investigations of phospholipid metabolism of leukocytes have been reported (6, 10) but we are not aware of similar studies on platelets.In this laboratory, investigation of the biochemical characteristics of leukemic leukocytes is being carried out, and as part of this program an examination of the phospholipid metabolism of these cells was undertaken. In this report are presented data on the phospholipid composition of platelets and leukocytes from leukemic patients, and the results of studies on the incorporation of P32-labeled orthophosphate (P32) into the phospholipids of these formed elements in vivo and in vitro.The phospholipid composition was similar in leukocytes and platelets from chronic leukemia and in leukocytes from acute leukemia. The pattern of incorporation into leukocyte phospholipids of p32 administered therapeutically to patients with chronic myelocytic leukemia was studied, and it was found that rapid incorporation of P32 could be demonstrated in lecithin, phosphatidylethanolamine, phosphatidylserine, inositol phosphatide, and sphingomyelin. In the course of this work it was * This study was supported in part by Research Grant E-2813 from the National Institute of Allergy and Infectious Diseases, Bethesda, Md. and by Research Grant P-143B from the American Cancer Society. A preliminary report of the work has been published (Fed. Proc. 1960, 19, 72 MATERIALS AND METHODSPaticets studied. Two patients with acute myelocytic leukemia, 2 with chronic lymphocytic leukemia, and 9 with chronic myelocytic leukemia were utilized in this study. Seven of the patients had not been treated previously for their leukemia. Six of the patients, all with chronic myelocytic leukemia, had been treated previously by chemotherapy or with P'3, but were in relapse with leukocyte counts above 100,000 per mm3 at the time of the study. There was no apparent difference in the results obtained from previously treated or untreated patients, although the series is too small to permit definite conclusions on this point.Separation of the formed elements of the blood. Leukocytes were isolated from patients with acute or chronic myelocytic or lymphocytic leukemia with white blood cell counts greater than 100,000 per mm3. Five hundred ml of blood wvas collected in a plastic platelet pack (Fenw-al) containing 50 ml of 3 per cent ethylenediamine tetraacetic acid (USP) as anticoagulant. All subsequent operations were performed at 40 C. The blood was stored for 1.5 hours to permit sedimentation of the erythrocytes. The plasma layer containing platelets and leukocytes was removed by syphoning into a plastic transfer pack (Fenwal) and then centrifuged in silicone-coated (G.E. Drifilm) 40 ml centrifuge tubes at 180 G for 15 minutes to sediment the leukocytes. The leukocytes were washed 3 times with twice their volume of 0.85 ...

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“…Quantitative studies on platelet lipids were done by Erickson, Williams, Avrin and Lee (33), who found that 16 per cent of the dry weight of platelets was lipid, 73 per cent of the lipid was phospholipid and 68 per cent of the phospholipid was "cephalin." Chargaff, Bancroft and StanleyBrown (34) reported that horse platelet "cephalin" shortened the recalcified clotting time of chicken plasma.…”

mentioning

confidence: 99%