William R. Vogler

A Controlled Study of the Efficacy of Granulocyte Transfusions in Patients with Neutropenia

A randomized clinical trial to determine the efficacy of granulocyte transfusions in neutropenic patients with infection was conducted. Criteria for patient selection included a proved infection, a granulocyte count of less than 300/mm3, availability of a suitable donor and failure to respond to at least 72 horus of appropriate antibiotic therapy. Thirty patients were assigned at random to receive either granulocyte transfusions or to serve as a control group. Antibiotic therapy was continued in both groups. Responses were judged by the degree of diminution of infectious episodes and survival. The results showed that 11 of 13 control patients failed to respond during the period of observation, whereas 10 of 17 patients given transfusions responded. The results were statistically significantly different (p less than 0.05). The median survival was 22.5 days in the group given transfusions (group 2) and 7.7 in the control group (group 1) (p less than 0.01). The granulocyte transfusions were most effective in patients with hypocellular marrows who failed to recover during the period of observation. These results indicate that granulocyte transfusions are effective in the short-term control of infections in neutropenic patients.

Effect of tamoxifen, a nonsteroidal antiestrogen, on phospholipid/calcium-dependent protein kinase and phosphorylation of its endogenous substrate proteins from the rat brain and ovary

Antiestrogens (tamoxifen, clomiphene and nafoxidine) were found to inhibit phospholipid/Ca2+-dependent protein kinase (PL/Ca-PK, or protein kinase C), whereas estrogens (estradiol and diethylstilbesterol) and the weakly estrogenic chlorotrianisene were inactive. Kinetic analysis indicated that the antiestrogens inhibited PL/Ca-PK competitively with respect to phosphatidylserine (Ki = 16-27 microM), but non-competitively with Ca2+ (Ki = 14-30 microM). Tamoxifen, but not diethylstilbesterol, also inhibited the phospholipid/Ca2+-dependent phosphorylation of various endogenous proteins from the total, solubilized fraction of the rat brain and ovary. Myosin light chain kinase, a calmodulin/Ca2+-dependent class of protein kinase, was similarly inhibited by tamoxifen; the drug, however, was without effect on cyclic AMP-dependent and cyclic GMP-dependent protein kinases. It is suggested that PL/Ca-PK, by virtue of the hydrophobic interactions required for the enzyme activation, may represent a potential site of action for the lipophilic antiestrogens, in addition to the commonly recognized intracellular estrogen receptors.

A phase III trial of recombinant human erythropoietin therapy in nonanemic orthopedic patients subjected to aggressive removal of blood for autologous use: dose, response, toxicity, and efficacy

BACKGROUND: Previous clinical trials have shown that the use of recombinant human erythropoietin (EPO) can facilitate autologous blood donation and reduce allogeneic blood transfusions in autologous blood donors who are anemic at first donation. However, the role of EPO therapy in nonanemic patients remains undefined. To identify this role, a randomized, controlled, multicenter dose‐escalation trial was conducted in patients whose initial hematocrit was > 39 percent (0.39).

Phospholipid-sensitive Ca2+-dependent protein kinase and its substrates in human neutrophils

Phospholipid-sensitive Ca2+-dependent protein kinase (PL-Ca-PK) was found to be present at a high level in human neutrophils, with its activity localized in the particulate fraction. In contrast, cyclic AMP-dependent protein kinase (A-PK) and cyclic GMP-dependent protein kinase (G-PK), present at lower levels compared to PL-Ca-PK, were localized in the cytosolic fraction. Phosphorylation of several endogenous proteins (mol. wts. 89,000, 38,000, 34,000, 17,000 and 15,000), also localized in the particulate fraction, was stimulated specifically by a combination of phosphatidylserine and Ca2+, whereas no substrate proteins were observed for the calmodulin-sensitive Ca2+-dependent protein kinase system under the same incubation conditions. Although no substrate proteins for G-PK were detected, one substrate (mol. wt. 19,000) for A-PK was observed. Phosphorylation of substrates for PL-Ca-PK, but not that for A-PK and for enzymes independent of Ca2+ or cyclic AMP, was inhibited by a variety of agents, including trifluoperazine, W-7 [N-(6-aminohexyl)-5-chloro-1-naphthalene-sulfonamide], adriamycin, palmitoylcarnitine, and melittin. The present findings suggest that the phospholipid/Ca2+-stimulated protein phosphorylation system may be important in the membrane associated functions of human neutrophils.

Bacterial endocarditis: A review of 148 cases

Abstract One hundred and forty-eight cases of bacterial endocarditis seen in the Emory University Medical School affiliated hospitals between January 1948 and April 1960 are reported. The age, sex, and race distribution and classification of infecting organisms are presented. No change in the relative frequency of occurrence of any of the organisms was found during the twelve years of this study. Recognition of endocarditis was the single most important factor affecting prognosis. The paucity of classic physical findings in patients with proved endocarditis is emphasized. Complications resulting from emboli were the primary factor in hospital mortality. The long-term prognosis of those patients with healed endocarditis was good; 82 per cent were alive five years after discharge from the hospital. Congestive heart failure was shown to be the major cause of death in those patients who survived the period of hospitalization. The time of appearance of symptoms, the mortality and the pathologic findings are shown to be a function of the virulence of the individual organism rather than an attribute of any particular group of organisms. A regimen of penicillin and streptomycin in dosages suited to the individual organism is proposed as the treatment of choice, with addition of vancomycin, erythromycin or chloramphenicol as needed for specific organisms.

Immunocytochemical evidence for phorbol ester-induced protein kinase C translocation in HL60 cells

The ability of tumor promoting 12-O-tetradecanoylphorbol-13-acetate (TPA) to redistribute protein kinase C in human promyelocytic leukemic HL60 cells was investigated. It was found that TPA caused a rapid translocation (within 10 min) of protein kinase C from the cytosolic (soluble) fraction to the particulate (membrane) fraction, as determined indirectly by assaying for the enzyme activity or by immunoblotting of the enzyme protein in the isolated subcellular fractions. Immunocytochemical localization of the enzyme demonstrated directly that the TPA caused an enzyme translocation t the plasma membrane. These findings suggest that translocation to the plasma membrane of the enzyme may represent initial events related to the TPA effect on terminal differentiation of HL60 cells to monocytes/macrophages.

The effect of recombinant human erythropoietin on the efficacy of autologous blood donation in patients with low hematocrits: a multicenter, randomized, double-blind, controlled trial.

Background: This randomized controlled study was undertaken to determine the effect of recombinant human erythropoietin (rHuEPO) on erythropoiesis, autologous blood collection, and allogeneic transfusion risk in elective surgery patients with low baseline hematocrits.

Bone Marrow Transplant Retinopathy

Five of eight patients (62%) who survived at least six months after autologous or allogeneic bone marrow transplantation for acute leukemia developed occlusive microvascular retinopathy. Treatable retinal microangiopathy included a high incidence (80%) of clinically significant macular edema and one case of proliferative retinopathy with subhyaloid hemorrhage. The bone marrow transplant protocol required high-dose cytarabine hydrochloride and 1,200 cGy of total body irradiation. The development of radiation retinopathy after such low doses of teletherapy suggests that high-dose chemotherapy may increase the susceptibility for the development of retinopathy at otherwise safe radiation doses.

Phospholipid-sensitive Ca2+-dependent protein kinase inhibition by R-24571, A calmodulin antagonist☆

R-24571 (calmidazolium), a derivative of the antimycotic agent miconazole, inhibited phospholipid-sensitive Ca2+-dependent protein kinase (PL-Ca-PK), with an IC50 (the concentration causing 50% inhibition) of 5.3 microM. It also inhibited the calmodulin/Ca2+-stimulated enzymes, with IC50 values of 1.6 and 0.1 microM for myosin light chain kinase (MLCK) and phosphodiesterase respectively. Analysis of inhibition by R-24571 of PL-Ca-PK and MLCK revealed complex kinetics, suggesting that the agent interacted with the cofactors, the enzyme, and/or the cofactor-enzyme complexes. At saturating concentrations of the cofactors, R-24571 inhibited PL-Ca-PK and MLCK noncompetitively with their respective cofactors. Inhibition of MLCK by R-24571 was completely overcome by phosphatidylserine, indicating a strong hydrophobic interaction between R-24571 and the phospholipid in the presence of calmodulin. R-24571 also inhibited phosphorylation of various endogenous proteins in brain stimulated specifically by phosphatidylserine/Ca2+ or calmodulin/Ca2+. The present findings inducated that R-24571 has little specificity in inhibiting two types of Ca2+-dependent protein kinases sensitive to phospholipid or calmodulin.

Correlation of cytosine arabinoside-induced increment in growth fraction of leukemic blast cells with clinical response.

The rate of 3H‐thymidine incorporation into DNA and labeling index were determined in fifteen adult patients with acute non‐lymphocytic leukemia by incubation of marrow aspirates in vitro with 3H‐thymidine before and 48 hours after receiving cytosine arabinoside. The cytosine arabinoside was administered using three different schedules, and was followed in 48 hours by high‐dose methotrexate plus Leucovorin and vincristine. The patients were treated with sequential courses of therapy, their clinical responses evaluated, and the results correlated with the rate of 3H‐thymidine incorporation into DNA and the labeling index obtained from marrow aspirates. Statistically significant differences in the initial DNA synthesis rate and increments thereof, and labeling index were found between those responding to chemotherapy and those not responding. High initial 3H‐thymidine incorporation and labeling index and the apparent ability to recruit additional cells into the growth fraction by chemotherapeutic manipulations seemed to define a population of leukemic patients susceptible to a cycle dependent regimen.