Marshall A. Lichtman

Reduced Red Cell Glycolysis, 2,3-Diphosphoglycerate and Adenosine Triphosphate Concentration, and Increased Hemoglobin-Oxygen Affinity Caused by Hypophosphatemia

Abstract A marked reduction in red cell glucose utilization, lactate production, and 2,3-diphosphoglycerate and adenosine triphosphate concentration occurred in a patient with intractable diarrhea ...

The paradox of the neutrophil's role in tissue injury

The neutrophil is an essential component of the innate immune system, and its function is vital to human life. Its production increases in response to virtually all forms of inflammation, and subsequently, it can accumulate in blood and tissue to varying degrees. Although its participation in the inflammatory response is often salutary by nature of its normal interaction with vascular endothelium and its capability to enter tissues and respond to chemotactic gradients and to phagocytize and kill microrganisms, it can contribute to processes that impair vascular integrity and blood flow. The mechanisms that the neutrophil uses to kill microorganisms also have the potential to injure normal tissue under special circumstances. Its paradoxical role in the pathophysiology of disease is particularly, but not exclusively, notable in seven circumstances: 1) diabetic retinopathy, 2) sickle cell disease, 3) TRALI, 4) ARDS, 5) renal microvasculopathy, 6) stroke, and 7) acute coronary artery syndrome. The activated neutrophilˈs capability to become adhesive to endothelium, to generate highly ROS, and to secrete proteases gives it the potential to induce local vascular and tissue injury. In this review, we summarize the evidence for its role as a mediator of tissue injury in these seven conditions, making it or its products potential therapeutic targets.

Rheology of Leukocytes, Leukocyte Suspensions, and Blood in Leukemia POSSIBLE RELATIONSHIP TO CLINICAL MANIFESTATIONS

Suspensions of leukemic lymphocytes and myeloblasts and blood of leukemic patients were studied to examine (a) the effect of leukemic cells on blood viscosity and (b) the ability of leukemic cells to traverse channels of capillary diameter. The viscosity of suspensions of leukemic cells was dependent logarithmically on (a) shear strain rate and (b) cytocrit, although, suspensions of small lymphocytes and of myeloblasts had a similar viscosity at equivalent shear rates and cytocrit. The minimum apparent viscosity (MAV) of leukemic cells and red blood cells, measured over shear rates of 2.3-230 s(-1) was dependent logarithmically on cytocrit. However, MAV was slightly greater for leukemic cells than for red cells at cytocrits up to 20%. At cytocrits above 20%. MAV of leukemic cells increased more rapidly than that of erythrocytes. For example, at a 15% cytocrit MAV(WBC) (1.85 centipoise) was only slightly greater than MAV(RBC) (1.59); whereas, at 45% cytocrit MAV(WBC) (14.9) was markedly greater than MAV(RBC) (3.81). The blood of subjects with leukemia with marked elevation of leukocyte concentration (leukocrits of 6-32%) had 24% higher mean MAV (3.72) than blood with a similar total cytocrit composed of red cells (3.00). A negative correlation was present between leukocrit and erythrocrit in chronic lymphocytic (r = - 0.82) and chronic granulocytic (r = - 0.81) leukemia. Therefore, the modest increase in whole blood MAV in leukemia can be explained by (a) the negative association of leukocrit and erythrocrit and (b) the rarity of leukocrits over 20% and total cytocrits over 45%. However, the MAV of blood of leukemic patients was 71% greater than expected on the basis of their packed red cell volume. Hence, the ratio of hemoglobin concentration (O(2) carrying capacity) to MAV was abnormally low in the subjects with leukemia studied. Individual leukemic leukocytes were nearly rigid. The mean deformability index (DI) of leukemic myeloblasts (1.22; 1.18) and lymphocytes (1.22; 1.40) as measured by filtration and elastometry, respectively, at 50 mm H(2)O negative pressure, approached that of a rigid body (1.0) as compared to red cells studied by filtration (3.09) or elastometry (4.23). The ability of leukemic cells to traverse nucleopore filter or micropipette channels was related to cell diameter. The relevance of the rheology of leukemic cells to the interruption of blood flow and of tissue oxygen delivery and thereby to clinical manifestations of leukemia is considered.

The paradox of the neutrophil’s role in tissue injury: a review

The neutrophil is an essential component of the innate immune system, and its function is vital to human life. Its production increases in response to virtually all forms of inflammation, and subsequently, it can accumulate in blood and tissue to varying degrees. Although its participation in the inflammatory response is often salutary by nature of its normal interaction with vascular endothelium and its capability to enter tissues and respond to chemotactic gradients and to phagocytize and kill microrganisms, it can contribute to processes that impair vascular integrity and blood flow. The mechanisms that the neutrophil uses to kill microorganisms also have the potential to injure normal tissue under special circumstances. Its paradoxical role in the pathophysiology of disease is particularly, but not exclusively, notable in seven circumstances: 1) diabetic retinopathy, 2) sickle cell disease, 3) TRALI, 4) ARDS, 5) renal microvasculopathy, 6) stroke, and 7) acute coronary artery syndrome. The activated neutrophilˈs capability to become adhesive to endothelium, to generate highly ROS, and to secrete proteases gives it the potential to induce local vascular and tissue injury. In this review, we summarize the evidence for its role as a mediator of tissue injury in these seven conditions, making it or its products potential therapeutic targets.

Improved methods for reducing calcium and magnesium concentrations in tissue culture medium: application to studies of lymphoblast proliferation in vitro

SummaryWe have compared several methods for reducing calcium and magnesium concentrations in tissue culture medium, with the objective of producing selective deficiency effects on the growth of mouse (L5178Y) and human (P1R) lymphoblasts. In experiments in which calcium- and magnesium-“free” McCoy’s medium was supplemented with 15% horse or fetal calf serum, enough calcium and magnesium was provided by serum to support normal lymphoblast growth rate. Either dialysis or chelating-resin treatment of horse or fetal calf serum reduced calcium and magnesium contents approximately 100-fold. Use of dialyzed sera resulted in reduced growth rate, although in most cases the reduction in growth could be attributed to other effects of dialysis on serum, inasmuch as growth in those experiments was not restored to normal by the addition of calcium and magnesium to the medium.In contrast, the reduction of lymphoblast growth rate that occurred when resin-treated serum was used was always attributable to removal of calcium and magnesium, as normal growth always occurred in cultures to which calcium and magnesium were added.To demonstrate a growth-inhibiting effect on either mouse or human lymphoblasts by severe reduction of either calcium or magnesium in the presence of normal amounts of the alternative cation, it was necessary to (a) expose McCoy’s Ca−Mg-“free” medium to chelating-resin to reduce further the residual cation concentrations; (b) wash cells from stock cultures in a medium devoid of calcium and magnesium prior to inoculation into experimental cultures; (c) reduce the proportion of serum in the final medium from 15 to 5%; and (d) add 100 μM EGTA to cultures. Under these conditions, growth of both cell types was completely abolished in the presence of normal magnesium but in the absence of added calcium, and markedly reduced in the presence of normal calcium but in the absence of magnesium. These modifications did not compromise growth in cultures containing normal concentrations of both ions.

Obesity and the Risk for a Hematological Malignancy: Leukemia, Lymphoma, or Myeloma

Studies on obesity and the risk for hematological malignancies are reviewed. The paper includes a discussion of the metabolic effects of obesity and their possible role in linking increased body fat to neoplasia.

Erythrocyte Adenosine Triphosphate Depletion during Hypophosphatemia in a Uremic Subject

Abstract In a patient with chronic renal disease and uremia who was under treatment with a low-protein diet, hemodialysis and aluminum hydroxide gel marked hypophosphatemia associated with anorexia...

Potasssium transport in human blood lymphocytes treated with phytohemagglutinin.

We have confirmed that phytohemagglutinin (PHA) rapidly enhances the uptake of potassium (K+) by human blood lymphocytes. PHA, however, did not produce an increase in lymphocyte K+ concentration. The apparent steady-state of cell K+ concentration despite the marked increase in uptake of 42K+ could be explained by either an increase in K+-K+ exchange or an increase in concentrative (active) K+ accumulation in association with an increase in the leak of K+ from the cell. We compared, therefore, the uptake of 42K+ with the decrement in cellular K+ content when active transport was inhibited by ouabain. These studies established that K+-K+ exchange was negligible in human blood lymphocytes and that the increase in 42K+ uptake after PHA treatment represented concentrative transport. Our studies did indicate that 42K+ exodus from PHA treated lymphocytes increased markedly from 19 to 38 mmol-1 cell water-1-h-1. Within the same time period K+ influx into PHA-treated lymphocytes increased from 20 to 38 mmol-1 cell water-1-h-1. Thus, PHA produces a marked increase in the permeability of the lymphocyte membrane to K+, and the increase in active K+ influx in PHA-treated lymphocytes may represent a homeostatic response by the membrane K+ transport system to the increase in K+ efflux. Increased K+ turnover was observed at the lowest concentrations of PHA which produced an observable increase in [3H]thymidine incorporation into DNA. Thus, PHA produces an increase in K+ permeability that closely parallels its mitogenic effect. The rapid increase in K+ influx preceding blastogenesis and mitogenesis is required, therefore, to maintain normal intracellular K+ concentration. An adequate intracellular K+ concentration is essential for the synthetic processes required for cell transformation or division.

Regulation of Sodium and Potassium Transport in Phytohemagglutinin-Stimulated Human Blood Lymphocytes

Phytohemagglutinin (PHA) or concanavalin A treatment of lymphocytes causes an increase in membrane permeability so that the leak rates of Na and K increase 1.5- to 2-fold. Active Na and K transport increase proportionately in response to the increased membrane permeability. We have examined the role of lymphocyte Na concentration in sustaining the increased Na and K transport observed after PHA treatment. Cell Na concentration increases from 14.8 to 20.5 mmol/liter cell water in PHA-treated lymphocytes (P < 0.001). Four lines of evidence suggest that the 5-6 mmol/liter cell water increase in lymphocyte Na accounts for the increase in active Na and K transport in mitogen-treated lymphocytes. First, PHA does not increase directly the maximal Na, K-ATPase activity of isolated lymphocyte membrane vesicles. Second, when the Na concentration is increased by 6 mmol/liter cell water in unstimulated lymphocytes, Na and K transport increase nearly twofold. Third, the cell Na concentration (15 mmol/liter cell water) is near the K(m) for Na activation of the Na, K-ATPase in lymphocyte membranes. The ATPase activity thus, is capable of increasing as the cell Na rises above normal. Fourth, if lymphocytes are incubated in a medium containing a low Na concentration, K transport does not maintain the internal K concentration and the fall in cell K is accentuated in PHA-treated lymphocytes. These studies indicate that the adaptive acceleration of Na and K transport in mitogen-treated lymphocytes is mediated by a small increase in cell Na.

Battling the Hematological Malignancies: The 200 Years' War

The delineation of the hematological malignancies began near the end of the first third of the 19th century with the recognition of the similarity among cases with lymph node tumors and an enlarged spleen (Hodgkins disease). Descriptions of chronic and acute leukemia and myeloma followed thereafter. In the first years of the 20th century the discovery of x-radiation permitted palliative orthovoltage radiation therapy of Hodgkins disease. Following World War II, legitimate drug therapy for the hematological malignancies was introduced: nitrogen mustard, adrenocorticotropic hormone and cortisone acetate, and anti-folic acid derivatives, initially aminopterin. Today, about 14 classes of drugs (different mechanisms of action) and >50 individual agents are being used, with others under study. Several examples of agents targeting specific transcription factors or oncoproteins have been introduced. Despite remarkable progress, including the ability to cure acute leukemia in about 70% of children, cure several genetic variants of acute myelogenous leukemia in younger adults, cure some cases of lymphoma in children and younger adults, and induce prolonged remission in many affected persons, the majority of patients face an uncertain outcome and shortened life. Thus, we have much to do in the next several decades. The significant hurdles we must overcome include: the apparent infrequency of an exogenous cause that can be avoided, the exponential increase in incidence rates with age and the dramatic negative effect of aging on the results of treatment, the challenge of one trillion or more disseminated cancer cells among which are a smaller population of cancer stem cells, the profound genetic diversity of the hematological malignancies (apparently hundreds of unique genetic primary lesions), the redundant growth and survival pathways defining the cancer phenotype, the decreasing market for pharmaceutical companies as therapy becomes more specific (fewer target patients) and drug development costs become more expensive, and the significant negative long-term effects of current therapy on both children and adults. These challenges will be gradually overcome, if we (a) develop new models of cooperation among academia, industry, and government, (b) continue the growth of international participation in cancer research (more keen minds to the task), and (c) convince the governments of the world, including that of the U.S., that an investment in minimizing the effects of cancer is as important as defending against other threats to the welfare and longevity of their citizens.