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J Levin

Publications and source records attributed to J Levin.

At least 181 records · Page 10Linked to original sources
PubMed1987

Thrombocytopoietic response to immunothrombocytopenia in nude mice.

Thrombocytopoiesis was evaluated in T cell-deficient nu/nu mice and in T cell-replete nu/+ controls to determine if abnormalities would be associated with the deficiency of T cells. Mice were studied in the unperturbed steady state and after acute immunothrombocytopenia was induced by an injection of guinea pig antimouse platelet serum (APS). The state of thrombocytopoiesis was determined from platelet counts, megakaryocyte size, megakaryocyte number, and numbers of Meg-CFC. Splenic lymphocytes were evaluated by response to the mitogens bacterial lipopolysaccharide (LPS), phytohemagglutinin (PHA), and concanavalin A (Con A). Hematocrits, reticulocyte counts, leukocyte counts, marrow cellularity, GM-CFC, and BFU-E also were measured. Steady state thrombocytopoiesis was identical in nu/nu and nu/+ mice. In response to an injection of APS, acute thrombocytopenia was followed by macromegakaryocytosis and rebound thrombocytosis in mice of both genotypes. Splenic Meg-CFC increased in nude mice after APS or an injection of normal guinea pig serum (NGpS), and splenic GM-CFC increased after APS. Neither Meg-CFC nor GM-CFC increased in the spleens of nu/+ mice, but they showed early transient increases in bone marrow that did not occur in nu/nu mice. Sporadic, but weak, mitogenic responses to PHA or Con A were occasionally observed with nu/nu spleen cells, but these did not correlate with the state of thrombocytopoiesis. The results demonstrated that platelet production was normal in nu/nu mice and that megakaryocytopoiesis and platelet production responded to the stimulus imposed by acute immunothrombocytopenia. Increases in megakaryocyte size and platelet production occurred independently of changes in numbers of Meg-CFC, GM-CFC, or BFU-E. A normal complement of T cells appears to be unnecessary for normal platelet production and its augmentation in response to the stimulus of acute immunothrombocytopenia in vivo.

Animals↗
PubMed1987

Regulation of thrombopoiesis: effects of the degree of thrombocytopenia on megakaryocyte ploidy and platelet volume.

We have established a murine model and techniques with which to serially study thrombocytopoiesis after induction of experimental immune thrombocytopenia of variable severity and duration. Bone marrow megakaryocyte ploidy distribution was determined by using unfractionated bone marrow, a polyclonal megakaryocyte-specific probe, and two-color, fluorescence-activated flow cytometry. With these techniques, the modal megakaryocyte ploidy class in normal murine bone marrow was 16N. Serial studies of bone marrow megakaryocyte ploidy after the induction of acute, severe thrombocytopenia (platelet count, less than 0.05 X 10(6) microL) demonstrated no detectable change in the ploidy distribution at 12, 24, and 36 hours after the onset of thrombocytopenia. At 48 hours, the modal ploidy class shifted from 16N to 32N, and the 64N class increased significantly (P less than .001). The ploidy distribution returned to normal 120 hours after the onset of thrombocytopenia. A lesser degree of thrombocytopenia (platelet count reduction to 0.100 to 0.200 X 10(6)/microL) delayed the modal ploidy class shift from 16N to 32N until 72 hours after the onset of thrombocytopenia. Chronic, severe thrombocytopenia (platelet count, less than 0.05 X 10(6)/microL for seven days) resulted in a modal ploidy class shift from 16N to 32N during the thrombocytopenic phase and an enhanced increase in the 64N megakaryocyte class during the recovery phase. Mean platelet volume (MPV) was simultaneously measured on isolated total platelet populations after induction of thrombocytopenia. MPV was significantly increased (P less than .001) as early as eight hours after the onset of acute, severe thrombocytopenia, 40 hours before a shift in the ploidy distribution. Mild thrombocytopenia (platelet count reduction to 0.400 X 10(6)/microL) was not associated with a ploidy shift but did result in a significantly increased MPV (P less than .001). These studies demonstrate that the temporal relationship and magnitude of the effects of thrombocytopenia upon megakaryocyte ploidy distribution are dependent upon the degree and the duration of the thrombocytopenic stimulus and that the effects of experimental thrombocytopenia on platelet volume and megakaryocyte ploidy are dissociated.

Acute Disease↗
PubMed1987

Multivariate flow cytometric characterization and enrichment of murine megakaryocyte progenitors (CFU-Meg).

Multivariate flow cytometric analysis and sorting of mouse bone marrow cells viably stained with Hoechst 33342 (HO) was utilized to obtain a subpopulation enriched at least 30- to 60-fold for clonogenic megakaryocyte precursors (CFU-Meg). HO fluorescence intensity, forward light scatter, and perpendicular light scatter were measured on each HO-stained bone marrow cell as it passed through a dual beam flow cytometer. Cells were sorted on the basis of their light scattering and HO fluorescence intensity properties. The sorting region which included cells with high forward light scatter, low perpendicular light scatter, and low (0.1-0.65 times the modal channel) HO fluorescence intensity contained approximately 0.6% CFU-Meg. Of all CFU-Meg, 75% were contained in the light scatter gate, and 56% of all CFU-Meg were in the low HO-sorting region. This technique offers a simple, one-step procedure to obtain preparations enriched in murine clonogenic megakaryocyte precursors.

Animals↗
PubMed1987

Ultrastructural analysis of murine megakaryocyte maturation in vitro: comparison of big-cell and heterogeneous megakaryocyte colonies.

Two morphologically distinct types of murine megakaryocyte (MK) colonies are present after three to seven days in soft agar culture: (a) "big-cell" colonies composed of ten to 30 large, mature-appearing megakaryocytes and (b) "heterogeneous" colonies consisting of approximately 100 or more cells at various stages of differentiation. Cytochemical and immunocytochemical techniques were used to study MK maturation in colonies as well as normal mouse bone marrow. Acetylcholinesterase (AChE), a specific marker for murine platelets and MK, was found in the perinuclear cisterna, endoplasmic reticulum, and occasionally, Golgi cisternae of MK in three-day big-cell colonies and immature bone marrow MK. MK in seven-day big-cell colonies and mature bone marrow MK showed additional reaction sites in the demarcation membrane system and occasional granules. In seven-day heterogeneous colonies, small cells resembled immature bone marrow MK with respect to AChE localization, whereas large cells corresponded to mature bone marrow MK. With immunogold procedures at the ultrastructural level, polyclonal antibodies against human platelet membrane glycoprotein IIIa and antimouse platelet antiserum labeled bone marrow MK and all MK from colonies grown in soft agar cultures for three to seven days. Granulocytes and macrophages in both bone marrow and soft agar cultures were negative for AChE and these immunocytochemical markers. These data indicate that the pattern of expression of AChE during maturation of MK is similar in vivo and in vitro and demonstrate, when using this marker at the fine-structural level, that a greater range of MK maturational stages is present in heterogeneous colonies than is observed in MK in big-cell colonies. Furthermore, we have confirmed that small cells in heterogeneous colonies are MK and that these colonies are composed solely of MK and their precursors.

Acetylcholinesterase↗
PubMed1986

Essential thrombocythemia and leukemic transformation.

Alkylating agents and 32P have been widely employed in the treatment of patients with essential thrombocythemia (ET). During a four-month period, we observed 3 cases of ET that had transformed into leukemia. Two patients had been treated with uracil mustard: One developed acute myelogenous leukemia 79 months after institution of therapy, and the other patient developed chronic myelomonocytic leukemia 24 months after the start of therapy. The third patient had been treated with busulfan, and ET evolved into myelofibrosis and eventually into acute undifferentiated leukemia with myelofibrosis. The patient who developed acute myelogenous leukemia was asymptomatic at the time of diagnosis of ET but was treated because his platelet count was greater than 1,000,000/mm3. He died 1 month after leukemic transformation, during induction chemotherapy. The other 2 patients presented with symptoms referable to their thrombocythemia. Review of the English literature revealed 12 other definite or probable cases of ET with leukemic transformation, all but 1 having been treated with alkylating agents and/or 32P. We propose that the natural history of ET may be similar to that of polycythemia vera, with evolution into leukemia being an unusual occurrence except in the setting of previous chemotherapy. Therefore, the current practice of treating asymptomatic patients with ET may not be justified, since administration of alkylating agents or 32P may increase the risk of subsequent development of leukemia.

Adult↗
PubMed1986

Immunohistochemical localization of membrane and alpha-granule proteins in plastic-embedded mouse bone marrow megakaryocytes and murine megakaryocyte colonies.

Using an immunoperoxidase technique that permits optimal antigen localization at the light microscope level, we have detected two platelet alpha-granule constituents and three platelet membrane glycoproteins in mouse bone marrow megakaryocytes and in murine megakaryocyte colonies grown in soft agar culture for three to seven days. Using polyclonal antibodies prepared against human platelet proteins, we have demonstrated labeling for von Willebrand factor, fibrinogen, and the membrane glycoproteins IIIa and GMP-140 in both bone marrow megakaryocytes and megakaryocyte colonies after seven days of culture. Using monoclonal antibodies to membrane glycoproteins IIb and GMP-140, we have demonstrated label in mouse bone marrow megakaryocytes. Granulocyte and macrophage colonies were negative for each of these markers. Murine bone marrow megakaryocytes and megakaryocyte colonies demonstrated a similar enzyme histochemical pattern: weakly positive for alpha-naphthyl acetate esterase and negative for chloroacetate esterase. These data indicate that megakaryocytes grown in soft agar culture express many of the same glycoproteins as bone marrow megakaryocytes. Furthermore, the ability of antibodies directed against human platelet membrane glycoproteins to identify murine megakaryocyte glycoproteins indicates that these constituents have been highly conserved during evolution.

Animals↗
PubMed1986

Plasma clearance rates of coagulation factors VIII and IX in factor-deficient individuals.

The plasma clearance rates of factors IX and VIII were determined in patients with hemophilia A and B who had received factor replacement by prolonged, continuous infusion of factor concentrates. The clearance rates were calculated by dividing the factor infusion rates by the steady-state plasma factor activities corrected for base-line factor activities. The mean factor IX clearance rate in eight factor IX-deficient patients was 233 mL/h (range 159 to 340 mL/h). The mean normalized clearance rate was 3.4 mL/h/kg. The mean factor VIII clearance rate in eight factor VIII-deficient patients was 294 mL/h (range 229 to 361 mL/h) and the mean normalized rate was 5.0 mL/h/kg. Both factors show linear relationships between the factor infusion rates and the steady-state plasma factor activities achieved.

Factor IX↗
PubMed1986

Association of decreased plasma cortisol and corticosteroid-binding globulin binding capacity in patients with essential hypertension and their first degree relatives.

Both plasma cortisol and total corticosteroid-binding globulin (CBG) binding capacity were lower in 210 patients with uncomplicated essential hypertension (EH) and their 66 hypertensive first degree relatives, compared with 171 controls. However, both variables were similar in 84 normotensive relatives to those found in control subjects showing extensive variation over the entire control-EH range. The association of reduced plasma cortisol and CBG binding capacity in EH may be closely related to altered steroid metabolism which may be partly explained by an abnormality mimicking a relative deficiency in adrenal 17 alpha- and 11 beta-hydroxylation. One manifestation of this disorder in EH is an increase in the secretion rate and plasma levels of dehydroepiandrosterone sulphate (DHEA-S) which is bound with high-affinity to a low-capacity specific plasma globulin and accounts for most of the digitalis-like activity of human plasma.

Adolescent↗
PubMed1986

Partial purification of thrombopoietin using lectin chromatography.

Thrombopoietin (TPO) was partially purified from the plasma of thrombocytopenic rabbits by ammonium sulfate fractionation and lectin chromatography. Thrombopoietic activity was measured with an in vivo assay that measures platelet production by the incorporation of 75Se-selenomethionine (75SeM) into newly forming platelets in mice. Thrombopoietin was precipitated from rabbit plasma by an ammonium sulfate saturation of 60%-80%. The ammonium sulfate precipitate was further fractionated by application to a column of wheat germ agglutinin (WGA) bound to agarose. The biological activity was eluted from the WGA column with 0.2 M N-acetylglucosamine. The minimum dose that stimulated thrombopoiesis was decreased 1000-fold to 1.3 microgram/g body weight by this first lectin step. The biological activity eluted from the WGA column was then applied to a column of concanavalin A (ConA) bound to agarose and eluted with 0.1 M alpha-methylmannose. The ConA step produced an additional two-fold increase in specific activity and decreased the minimum dose to 0.65 microgram/g body weight. Stimulation of thrombopoiesis was not associated with an increased peripheral platelet count. Although the purification scheme presented provided a 7000-fold increase in specific activity compared to plasma from thrombocytopenic rabbits, analysis of the WGA and ConA fractions by gel-permeation high-performance liquid chromatography (GP-HPLC) and SDS-PAGE indicated that many proteins were still present at this stage of the purification procedure.

Animals↗
PubMed1985

Technetium-99m glucoheptonate as a scanning agent in hepatocellular carcinoma.

Technetium-99m glucoheptonate (Tc-99m GH) is concentrated in pulmonary and cerebral tumors. The purpose of this study was to assess the uptake of this radionuclide by hepatocellular carcinoma. Its concentration by the primary tumor was compared with that in the non-neoplastic hepatic tissue in 31 patients who showed obvious defects on a colloid scan, and its uptake by pulmonary metastases was examined in six patients with x-ray evidence of this complication. In two patients, the uptake by the tumor was greater than, in six it was equal to, and in ten it was less than that in the non-neoplastic hepatic tissue. In the remaining 13 patients, there was no concentration at all in the tumor. In none of the six patients with multiple pulmonary metastases could uptake of Tc-99m GH by the metastases be demonstrated. It is concluded that Tc-99m GH is of limited value in the diagnosis of primary or metastatic hepatocellular carcinoma.

Adult↗
PubMed1985

The clinical value of scintigraphic brain scanning. Experience at the Hillbrow Hospital, Johannesburg, South Africa.

Patients were referred to the Department of Nuclear Medicine for brain scintigraphy to be screened for possible intracranial pathology. These referrals were made in order to reduce the heavy load on the transmission computerized tomography (TCT) facilities. Great clinical importance, therefore, has been attached to scintigraphic findings; this emphasizes the need for an accurate assessment of the predictive value of this procedure.

Adolescent↗
PubMed1985

The history of the development of the Limulus amebocyte lysate test.

The Limulus Test for bacterial endotoxins was developed as the result of investigations of the effects of Gram-negative bacteria on blood coagulation in the horseshoe crab Limulus polyphemus. Analysis of the factors required for blood coagulation (all of which are contained within Limulus amebocytes) has revealed marked similarities between blood coagulation in the horseshoe crab and mammals. The Limulus Test (LT) is the most sensitive test for bacterial endotoxins now available and correlates with a wide variety of other assays. An increasing number of applications of the LT have been described, including detection of endotoxins in pharmaceutical preparations, food, radioisotopes, water supplies, and medical devices, as well as in various body fluids such as blood, cerebrospinal fluid, and urine. The "basic research" that unexpectedly led to the discovery and subsequent development of the Limulus Test provides an example of the value of biomedical investigations directed at understanding pathophysiologic phenomena and demonstrates the importance of studies of comparative physiology.

Animals↗