Dorothea Zucker-Franklin

The characterization of soluble amyloid prepared in water

Amyloid was extracted from the spleen of a patient with primary amyloidosis by homogenizing it at high speed with water after preliminary treatments, first to remove proteins soluble in saline, and then to remove salts. The extracts containing amyloid appeared to be clear at concentrations up to 6 mg/ml of protein. The material gave little sediment on being centrifuged up to 20,000 g for 1 hr, but the protein was sedimented at 100,000 g in 1 hr. The amyloid could be precipitated from the extracts by addition of NaCl to 0.0075 mole/liter or of CaCl(2) to 0.0025 mole/liter. The protein-bound Congo red formed a red precipitate and this property was used to estimate recovery and purity of amyloid during extraction. On electronmicroscopy the isolated amyloid proved to be morphologically pure. It existed either as single filaments measuring 60-80 A in diameter or as large aggregates of these filaments.Freshly isolated amyloid in water sedimented as a single homogeneous peak with an s degrees (20,[unk]) of about 45-50S. On standing, the solution became cloudy and more rapidly sedimenting components appeared. On electrophoresis the material migrated as a homogeneous peak towards the anode. The protein had an amino acid composition different from that of all known serum proteins. It was rich in acidic amino acids and had little cysteine and methionine and no hydroxyproline. The total content of carbohydrate was less than 2%.

Leukemic cells with membrane properties of thymus-derived (T) lymphocytes in a case of Sézary's syndrome: Morphologic and immunologic studies

Abstract The abnormal lymphocytes of a patient with Sezarys syndrome have membrane markers which differ from those recognized on other leukemic cells. They form rosettes with sheep erythrocytes and lack complement receptor sites and surface immunoglobulin. We show that this combination of markers is possessed by a distinct population of normal lymphocytes, which evidence indicates are T cells.

Microfibrils of blood platelets: their relationship to microtubules and the contractile protein

Human blood platelets were subjected to osmotic shock, brief sonication, pressure homogenization, or treatment with adenosine diphosphate (ADP). These procedures demonstrated an abundance of cytoplasmic microfibrils. The fibrils resembled those found on electron microscopy of partially purified thrombosthenin, the actomyosin-like protein isolated from platelets, and they also appeared to resemble the myofilaments of smooth muscle. Similar fibrils were not found in leukocytes studied under identical conditions. Treatment with colchicine (2 x 10(-5) mole/liter) resulted in the disappearance of microtubules but did not affect the morphology of the microfibrils or interfere with platelet-dependent clot retraction. Thus, microfibrils rather than microtubules may represent the morphologic counterpart of the contractile protein. Brief osmotic shock at low temperature or treatment with 10(-4) M ADP caused the marginal band of microtubules to be replaced by a bundle of intertwining microfibrils. The apparent inter-conversion of microtubules and microfibrils under a variety of conditions led to the hypothesis that fibrils and tubules consist of similar subunits whose degree of polymerization might be dependent on local cytoplasmic forces. Furthermore, on the basis of these observations, it is postulated that the contractile properties of the cells may be vested in the microfibrils, whereas the tubules may serve to maintain the highly asymmetric shape characteristic of circulating and irreversibly aggregated platelets.

Red-Cell and Platelet Fragmentation in Idiopathic Autoimmune Thrombocytopenic Purpura

We investigated the abnormal small-particle spike discerned in the platelet-rich plasma of patients with severe idiopathic autoimmune thrombocytopenic purpura. By electron microscopy, erythrocyte as well as platelet fragments were found in the 27,000 X g plasma sediment of 15 patients with severe disease. These fragments were not observed in the plasma sediment of 12 normal subjects, two healthy asplenic subjects, three patients with thrombocytopenia of nonimmunologic origin, and two with autoimmune thrombocytopenic purpura in remission. Weak complement sensitization of red blood cells was noted in seven out of 12 patients with the disease. Coating of red blood cells with IgG or IgM was not detected in these patients. Whereas erythrophagocytosis was conspicuously absent, phagocytosis of intact platelets as well as platelet fragments and other cellular debris was frequently observed. Autoimmune mechanisms may be directed against erythrocytes as well as platelets in most cases of severe idiopathic autoimmune thrombocytopenia.

The ultrastructure of cells in human thoracic duct lymph

The cells of human thoracic duct fluid were concentrated and studied with the help of the electron microscope. Though most of the cells were considered to by typical lymphocytes, there proved to be a marked heterogeneity within this cell population. Some cells were completely devoid of rough endoplasmic reticulum whereas in others this organelle was well developed. This finding is discussed in the light of recent evidence that thoracic duct lymphocytes may play an important role in the immune response. The high incidence of multivesicular bodies and the occurrence of fibrillar structures in these cells is also pointed out. In addition it was observed that many thoracic duct eosinophiles have a paucity of specific granules and a somewhat degenerated appearance. The absence of neutrophiles, basophiles, and thrombocytes in thoracic duct lymph was also noted.

Microthrombocytosis and Platelet Fragmentation Associated with Idiopathic/Autoimmune Thrombocytopenic Purpura

Platelet volume distribution curves were obtained in 20 control subjects and in 21 patients with idiopathic/autoimmune thrombocytopenic purpura. A striking increase in microthrombocytes as well as megathrombocytes was noted in 86% of patients on one or more occasions, particularly in the presence of severe thrombocytopenia. The entire spectrum of platelet volume distribution curves noted in patients could be reproduced experimentally in rabbits following intravenous injection of anti‐platelet antibody. Differential centrifugation studies with control subjects revealed that microthrombocytes were light platelets and megathrombocytes were heavy platelets. Electron microscopy in patients with thrombocytopenia revealed that microthrombocytes were composed of intact small platelets as well as platelet fragments. It is concluded that severe peripheral destruction of platelets is associated with an increase in microthrombocytes as well as megathrombocytes.

Current concepts of amyloid.

Publisher Summary This chapter discusses the structure, chemistry, immunology, cellular origin, metabolism, and possibly the pathogenesis of amyloid. The simplest and most widely used classification of amyloidosis is based on the four major categories introduced by Reimann. Primary amyloid occurs with no known antecedent or coexistent disease and usually involves mesodermal tissues, such as smooth or skeletal muscle, or the cardiovascular system. Generally there is variability in staining of the deposits with Congo red, iodine, and metachromatic dyes. Secondary amyloid is usually associated with chronic diseases, such as infections, neoplasms, neurological disorders, or connective tissue diseases—especially rheumatoid arthritis. It generally involves spleen, liver, kidney, intestines, and adrenals and tends to have reproducible and characteristic staining properties with the above mentioned dyes. Amyloid associated with myeloma tends to resemble the primary type but is invariably associated with a neoplasm involving plasma cells or lymphocytes—such as multiple myeloma, macroglobulinemia, or heavy chain disease. Tumor-forming amyloid is characterized by the small masses of amyloid in the skin, eye, bladder, urethra, respiratory tract, or other organs and is generally unassociated with any underlying disease.

Ultrastructure of thrombosthenin, the contractile protein of human blood platelets.

Partially purified thrombosthenin with adenosine triphosphatase activity similar to that of actomyosin was subjected to electron microscopy. More than 50 percent of the material consisted of fibrils 80 to 100 angstroms in width. Occasional fibrils suggested a periodic structure. The morphology of isolated thrombosthenin resembled that of microfibrils in the cytoplasm and pseudopods of intact platelets.

The identification of eosinophil colonies in soft-agar cultures by differential staining for peroxidase.

There has been a need to easily quantitate the incidence of eosinophil colonies within soft agar cultures. This has been realized by layering of the agar with benzidine dihydrochloride that permits detection of peroxidase activity in cells. Eosinophil colonies can be specifically identified by the addition to the substrate of potassium cyanide, an inhibitor of enzyme activity in neutrophils and monocytes. The enumeration of eosinophil colonies can be accomplished by scanning fresh or embedded cultures with low power magnification.

HTLV tax and Mycosis Fungoides

To the Editor: Although a causal relation between human T-cell lymphotropic retrovirus type I and type II (HTLV-I and HTLV-II) and adult T-cell leukemia has been well established, the association b...