Treatment of severe acute lung allograft rejection with OKT3 and temporary extracorporeal membrane oxygenation bridging.

OBJECTIVES: The use of OKT3 for treatment of advanced high-grade acute rejection episodes eventually can result in cytokine release and consecutive pulmonary edema. Temporary extracorporeal membrane oxygenation (ECMO) bridging can be used to overcome this crucial period before the beneficial effects of OKT3 can be observed. METHODS: We summarize our experience with three patients, who underwent lung transplantation and presented with severe acute rejection episodes. OKT3 had to be initiated due to insufficient response to standard rejection therapy with corticosteroids. Upon initiation of OKT3 treatment, a massive life-threatening deterioration of lung function in spite of heavily invasive respirator treatment was seen and temporary ECMO support was imperative to support graft function. Results of this treatment were retrospectively reviewed. RESULTS: In all cases femoro-femoral veno-arterial ECMO was used for support of the impaired graft and after a period of 4-5 days led to a massive improvement of graft function. In the further course two patients could be discharged from hospital and are still alive 30 and 36 months, respectively, after the described incident. One patient died 4 months later due to liver failure. CONCLUSIONS: We conclude that the use of ECMO support in patients experiencing significant side effects from OKT3 therapy is a useful and effective therapeutic tool to overcome the initial critical period until the lung has sufficiently recovered.     

Purification and properties of a fibrinolytic enzyme from Bacillus subtilis

A fibrinolytic enzyme obtained from B. subtilis was purified, using DEAE-cellulose column chromatography, and gel filtration on Sephadex G-100. The preparation was homogeneous as tested by gel filtration on Sephadex G-200, and disc electrophoresis. The molecular weight of this enzyme was 29.400 estimated by gel filtration on Sephadex G-100. The optimum pH for enzyme activity was 7.2. Copper ions significantly increased enzyme activity, while Zn⁺⁺ and Mn⁺⁺ caused marked inhibition.     

Changes in lipid profile variables in response to submaximal and maximal exercise in trained cyclists

This study examined the effect of prolonged submaximal exercise followed by a self-paced maximal performance test on cholesterol (T-Chol), triglycerides (TG), and high-density lipoprotein cholesterol (HDLC). Nine trained male athletes cycled at 70% of maximal oxygen consumption for 60 min, followed by a selfpaced maximal ride for 10 min. Venous blood samples were obtained at rest, at 30 and 60 min during submaximal exercise, and immediately after the performance test. Lactic acid, haematocrit (Hct), haemoglobin (Hb), T-Chol and TG were measured in the blood, while plasma was assayed for HDL-C. Plasma volume changes in response to exercise were calculated from Hct and Hb values and all lipid measurements were corrected accordingly. In order to ascertain the repeatability of lipid responses to exercise, all subjects were re-tested under identical testing conditions and experimental protocols. When data obtained during the two exercise trials were analysed by two-way ANOVA no significant differences (P > 0.05) between tests were observed. Consequently the data obtained during the two testing trials were pooled and analysed by one-way ANOVA. Blood lactic acid increased non-significantly (P > 0.05) during the prolonged submaximal test, but rose markedly (P < 0.05) following the performance ride. Lipid variables ascertained at rest were within the normal range for healthy subjects. ANOVA showed that blood T-Chol and TG were unchanged (P > 0.05), whereas HDL-C rose significantly (P < 0.05) in response to exercise. Post hoc analyses indicated that the latter change was due to a significant rise in HDL-C after the performance ride. It is concluded that apparent favourable changes in lipid profile variables occur in response to prolonged submaximal exercise followed by maximal effort, and these changes showed a good level of agreement over the two testing occasions.     

Shear‐Induced Crystallization and Shear‐Induced Dissolution of Poly(ethylene oxide) in Mixtures with Tetrahydronaphthalene and Oligo(dimethyl siloxane‐b‐ethylene oxide)

Cloud point temperatures (T_cp) and crystallization temperatures (T_l/s) were measured at different constant shear rates for the ternary system tetrahydronaphthalene/poly(ethylene oxide)/oligo(dimethyl siloxane‐b‐ethylene oxide) using a rheo‐optical device and in the case of T_l/s additionally a viscometer. This system enables for the first time a joint investigation of both transitions with a given mixture. Shear favors the homogeneous liquid state and the formation of crystals. T_cp (liquid/liquid demixing, UCST) shifts to lower and T_l/s (liquid/solid, segregation of PEO) to higher temperatures by several degrees as the shear rate, , is increased up to 500 s^?1. The normalized shift in T_cp fits well into previous results for high molecular weight blends, oligomer mixtures, polymer solutions in single solvents and low molecular weight mixtures. A phase separated near critical blend was examined 1 K below its T_cp by means of a shear cell (Linkam) in the quiescent state and under shear with respect to its morphology. Upon an increase in one observes a transition from the co‐continuous structures existing in the quiescent state via deformed and oriented particles to string like morphologies. Finally, at sufficiently high shear rates the mixture becomes homogeneous and structures can no longer be seen under the microscope. The morphologies developing after the secession of shear are pointing to pronounced influences of the flow history of the system on the final structure of two phase blends.

Equilibrium phase diagram of the system THN/COP/PEO at the indicated temperatures as obtained from turbidimetric titration. The curve for 42 °C indicates the compositions under which the mixtures segregate the first solid PEO particles upon cooling. The curves for the higher temperatures denote the demixing of the homogeneous system into two liquid phases.     

Fluorescent in situ hybridization study of c-myc oncogene copy number in prostate cancer

We previously conducted a study of 88 cases of prostate cancer in an attempt to identify potential prognostic biomarkers that can distinguish aggressive cases that must be treated immediately. Prostate cancer is a serious disease affecting men worldwide and compromises the quality of life of its patients. Biomarkers studied included chromosome 7 trisomy, chromosome 8 trisomy, and HER-2/neu oncogene amplification. These biomarkers were initially studied because trisomy 8 and oncogene amplification of the HER-2/neu gene have been reported in many other cancers, including those studied in this laboratory. In view of the fact that HER-2/neu amplification was not found to play a prominent role in the group of prostate cancer specimens that we studied, an exploration of other biomarkers was felt to be warranted. Thus, we began a pilot study of c-myc oncogene copy number in prostate cancer using the same protocol for fluorescent in situ hybridization and a direct-labeled SpectrumOrange LSI c-myc probe (Vysis, Inc., Downers Grove, IL) on formalin-fixed, paraffin-embedded tissue. From a total of 36 cases of prostate cancers successfully analyzed, we found 11 (31%) tumors exhibiting 3 or more positive signals for c-myc in 15% or more of the cells. Of these, only 7 tumors (19% of the total cases studied) had >/=3 signals in 20% or more of the cells. No case had >/=3 signals in 25% or more of the cells. Compared to other molecular probes tested, the c-myc signals were more faint and the quality of the preparation was less optimal than other tumor specimens that we previously studied. Based on the information available thus far, we conclude that an increased copy number in c-myc oncogene copy number was not a prominent finding in our cohort of prostate cancer patients.     

Phase I and pharmacokinetics studies of prochlorperazine 2-h i.v. infusion as a doxorubicin-efflux blocker

In an earlier phase I study, we reported that the maximal tolerated dose (MTD) of prochlorperazine (PCZ) given as a 15-min i.v. infusion was 75 mg/m². The highest peak plasma PCZ concentration achieved was 1100 ng/ml. The present study was conducted to determine if PCZ levels high enough to block doxorubicin (DOX) efflux in vitro could be achieved and sustained in vivo by increasing the duration of i.v. infusion from 15 min to 2 h. The treatment schedule consisted of i.v. prehydration with at least 500 ml normal saline (NS) and administration of a fixed standard dose of 60 mg/m² DOX as an i.v. bolus over 15 min followed by i.v. doses of 75, 105, 135, or 180 mg/m² PCZ in 250 ml NS over 2 h. The hematologic toxicities attributable to DOX were as expected and independent of the PCZ dose. Toxicities attributable to PCZ were sedation, dryness of mouth, anxiety, akathisia, hypotension, cramps, and confusion. The MTD of PCZ was 180 mg/m². Large interpatient variation in peak PCZ plasma levels (91–3215 ng/ml) was seen, with the plasma half-life (t1/2) being approximately 57 min in patients given 135–180 mg/m² PCZ. The volume of distribution (V_d), total clearance (Cl_T), and area under the curve (AUC) were 350.1±183.8 l/m², 260.7±142.7 l m² h^–1 and 1539±922 ng ml h^–1, respectively, in patients given 180 mg/m² PCZ and the respective values for patients receiving 135 mg/m² were 48.9±23.76 l/m², 33.2±2.62 l m² h^–1, and 4117±302 ng ml h^–1. High PCZ plasma levels (>600 ng/ml) were sustained in all patients treated with 135 mg/m² PCZ for up to 24 h. DOX plasma elimination was biphasic at 135 and 180 mg/m² PCZ, and a>10-ng/ml DOX plasma level was maintained for 24 h. Partial responses were seen in three of six patients with malignant mesothelioma, in two of ten patients with non-small-cell lung carcinoma, and in the single patient with hepatoma. Our data show that PCZ can be safely given as a 2-h infusion at 135 mg/m² with clinically manageable toxicities. The antitumor activity of the combination of DOX and PCZ needs to be confirmed in phase II trials.This work was supported by NIH grant R01 CA-29360 and S1488, CRC grant M01 RR-05280, and the Joan Levy Cancer Foundation. This paper was presented at the meeting of the American Association for Cancer Research, Orlando, Florida, May 19–22, 1993