Progressing prostate carcinoma.

In the Karnell Cancer Center Grand Rounds, we present a patient who underwent radical prostatectomy with bilateral pelvic lymphadenectomy, but had positive margins and subsequently developed local recurrence and then systemic disease. Pathologic and radiologic aspects of his disease are discussed. Therapeutic options at different stages of the disease are examined from the point of view of the urologist, radiation oncologist, and medical oncologist. The surgical portion of the discussion focuses on the selection of initial therapy. Both the selection of surgical candidates and choice of pre- or post-operative therapy in patients can be aided by prognostic tools looking at several variables, including prostate-specific antigen (PSA) level, Gleason score of the tumor, seminal vesicle invasion, extracapsular invasion, and lymph node involvement. Low-risk patients can be treated with monotherapy, such as radical prostatectomy, external beam radiation therapy, prostate brachytherapy, or cryosurgical ablation of the prostate. Higher risk patients may require adjuvant and possibly neoadjuvant therapy in addition. The radiation portion of the discussion focuses on the use of radiation therapy as salvage for relapsing disease. Of particular importance is the point that treating high-risk patients whose PSA levels have started to rise but are less than 1 ng/ml results in a long-term PSA control rate as high as 75%, but that limiting the use of salvage radiation therapy to patients with high PSA levels or biopsy confirmation of local recurrence in the face of a negative bone scan results in biochemical long-term control of less than 40%. In the medical oncology part of the discussion, the major focus is on the use of chemotherapy to treat patients whose disease has become resistant to hormonal therapy. Mitoxantrone plus a corticosteroid has been found to offer significant palliation for such patients. Combination therapy with estramustine plus taxanes, other microtubule inhibitors, or other agents such as topoisomerase II inhibitors, has been found to cause shrinkage of measurable soft tissue disease and diminution of serum PSA levels. The development of effective hormonal and chemotherapeutic drugs for treatment of metastatic disease has led to new interest in adjuvant and neoadjuvant therapy of high-risk patients.

Noninvasive prenatal diagnosis. Use of density gradient centrifugation, magnetically activated cell sorting and in situ hybridization.

OBJECTIVE: To develop a noninvasive method suitable for clinical prenatal diagnosis. STUDY DESIGN: Fetal nucleated erythrocytes were separated from peripheral blood of 17 healthy pregnant women using small magnetically activated cell sorting columns (MiniMACS) following density gradient centrifugation and dual antibody labeling methods. The protocol was designed to compare the efficacy of antitransferrin receptor (CD71)/antiglycophorin A (GPA) antibodies with antithrom-bospondin receptor (CD36)/anti-GPA antibodies in identifying nucleated erythrocytes in maternal blood. Cytospin preparations of the isolated cells were subjected to in situ hybridization with specific DNA probes for the Y chromosome and chromosome 21 to confirm the fetal origin. RESULTS: After MiniMACS the enrichment factors for the CD71/GPA- and CD36/GPA-positive cells from maternal blood were similar, and the percentages of fetal cells recovered did not differ. Seven of seven male pregnancies were correctly identified. One case of trisomy 21 was detected. CONCLUSION: The in situ hybridization analysis of fetal nucleated erythrocytes isolated from maternal blood using single density gradient centrifugation, anti-CD71/anti-GPA immunostaining and MiniMACS could be an accurate, sensitive and noninvasive method for prenatal diagnosis.

Platelet disorders in pregnancy: implications for mother and fetus.

Platelet disorders in pregnancy are not uncommon. Most often, obstetricians are faced with a patient with undiagnosed thrombocytopenia and have the responsibility of deciding if the condition is ITP, gestational thrombocytopenia, or a process related to pre-eclampsia. Correct diagnosis is important because ITP can be associated with fetal thrombocytopenia, making route of delivery important. In alloimmune thrombocytopenia, the mother develops antibodies to a specific platelet antigen present on the fetal platelet but absent on her own. Severe AIT can cause intracranial hemorrhage and have disastrous consequences for the fetus and neonate. In preliminary studies antenatal intravenous gamma globulin therapy has shown promise in preventing the development of intracranial hemorrhage and ameliorating fetal thrombocytopenia. Essential thrombocythemia with a platelet count of greater than 600 x 10(9) platelet/L can occur in pregnancy; therapy consists of antiplatelet aggregating agents such as aspirin, and plateletpheresis. Platelet function disorders can be acquired or inherited. Acquired platelet dysfunction disorders are usually caused by drugs such as aspirin or indomethacin, or by a systemic disease. Hereditary disorders of platelet function can be diagnosed in utero, but cordocentesis may represent an unacceptably high risk. For prenatal diagnosis, other methods, such as chorionic villus sampling or amniocentesis, should be investigated as an alternative to the potentially high risk of cordocentesis.

Thromboembolic complications in patients with advanced cancer: anticoagulation versus Greenfield filter placement.

Thirty patients with Stage III/IV cancer and thromboembolic complications between 1987-89 were reviewed. Twelve patients had a deep venous thrombosis proximal to the calf diagnosed by duplex scanning or contrast venography, 15 patients had a pulmonary embolism diagnosed by a high-probability pulmonary ventilation/perfusion scan or arteriogram, and three patients had both deep vein thrombosis and pulmonary embolism. Patients were treated primarily with anticoagulation (Group A = 20 patients) or a Greenfield filter (Group B = 10 patients). Seventy-five percent (15/20) of the Group A patients developed 19 bleeding or thrombosis-related complications: major bleeding (7), recurrent deep venous thrombosis/pulmonary embolism (4), inability to attain consistent therapeutic anticoagulation levels (3), heparin-induced thrombocytopenia (3), or progression of deep vein thrombosis (2). A Greenfield filter was eventually placed in 10 (50%) of the Group A patients without complications. Thirty percent (3/10) of the Group B patients developed progression of deep vein thrombosis that required anticoagulation. One other Group B patient died due to a guidewire-induced arrhythmia. Although patients with advanced cancers and venous thromboembolic disease have a high complication rate with either treatment, initial treatment with a Greenfield filter appears more definitive. Anticoagulation should be reserved for patients with progressive, symptomatic deep vein thromboses after placement of a filter.

The in vitro effects of selected environmental toxicants on two heme synthesis enzymes.

Benzene and some of its substitution products become environmental toxicants due to improper disposal procedures. Benzene has been found to alter heme and globin synthesis in anucleate rabbit reticulocytes (Forte et al., 1976; Wildman et al., 1976) and based on these findings we felt it would be useful to determine what, if any, effect these derivatives would have on heme synthesis in vitro by studying their influence on delta-aminolevulinic acid synthetase (ALAS) and ferrochelatase (FC) activities in rat liver homogenates. ALAS was measured according to Ebert et al. (1970). FC was measured after Williams et al. (1980). Final concentrations of each added compound to the reaction mixture were 10(-3) to 10(-6) M. Normal values for rat liver ALAS were 250-350 nmol ALA/g protein/30 min, mean 290 +/- 40, and for FC were 12-40 mumol heme/g protein/45 min, mean 20 +/- 7. At 10(-3) M and lower concentrations these compounds inhibited ALAS and stimulated FC activities. Their effect on ALAS activity expressed as percentage of control of three analyses performed in triplicate +/- SEM was: o- and p-dinitrobenzenes-46 +/- 2; trinitrotoluenes-55 +/- 2; dinitrotoluenes-70 +/- 2; and amino-dinitrotoluenes-171 +/- 4. The stimulatory effect of these compounds expressed as percentage of control +/- SEM on FC was: dinitrotoluenes-171 +/- 3; dinitrobenzenes-152 +/- 3; trinitrotoluenes-142 +/- 4; and amino-dinitrotoluenes-130 +/- 4. Other classes of compounds tested did not significantly affect these enzymes at the same concentrations. These in vitro techniques may prove useful for predicting in vivo toxicologic effects of pollutants on species of interest.

Natural history of potassium-deficiency myopathy in the dog: role of adrenocorticosteroid in rhabdomyolysis.

Potassium deficiency occurs in several conditions and is reported to cause muscle weakness and rhabdomyolysis. The mechanisms by which potassium deficiency cause muscle disease remain unknown, but the primary purpose of the present study was to determine whether abnormal muscle glycogen metabolism causes muscle weakness, as suggested by previous work. We monitored the natural history of potassium deficiency in two groups of dogs, one of which also received deoxycorticosterone acetate (DOCA), an agent commonly used in other studies to accelerate potassium loss. Group I dogs on potassium-free diet alone showed a 41% decrease in muscle potassium, no change in serum CO2, creatine kinase (CK), or muscle phosphorylase activity and only mild histopathologic abnormalities before death, after 198 +/- 42 days on the diet (mean +/- S.D.). In contrast, group II dogs on the same diet plus DOCA developed clinically similar severe weakness and died more rapidly than group I, 37 +/- 7 days (p less than 0.03). DOCA dogs showed a more rapid decrease in muscle potassium to the same level as group I, a 37% increase in serum CO2, an increase in serum CK to 1060 to 2775 IU/ml, a 23% decrease in muscle phosphorylase activity, and severe muscle histopathology, including rhabdomyolysis. Neither group showed any change in body weight, electromyogram (EMG), muscle glycogen concentration, glycogen synthetase activity, serum or muscle magnesium or phosphorus, or serum T3 or T4. In conclusion, dietary potassium deficiency in dogs causes severe weakness and death without causing rhabdomyolysis or abnormal muscle glycogen metabolism. Adding DOCA to the potassium-free diet creates a different model characterized by rapid clinical deterioration and rhabdomyolysis.

Improved radiochemical method for measuring ferrochelatase activity.

We describe a radiochemical method for measuring ferrochelatase activity in the in vitro incorporation of iron into mesoporphyrin-IX to form mesoheme. 59Fe is used to quantify ferrochelatase activity in rat liver. The uptake of iron is linearly related to the time allowed for it to occur, and the reaction proceeds optimally under reducing and anerobic conditions, maintained in sealed lyophilization vials under a positive pressure of nitrogen.

Alterations in platelet surface sialytransferase activity and platelet aggregation in a group of cancer patients with a high incidence of thrombosis.

Platelet aggregation, platelet surface sialic acid, and platelet surface sialytransferase activity were studied in a group of 12 cancer patients with a high incidence of thrombosis. These patients demonstrated accelerated coagulation, increased Factor VIII antigen and restocetin cofactor, and enhanced adenosine 5'-diphosphate-induced platelet aggregation. Platelet exogenous sialytransferase activity was increased in cancer patients (117.6 +/- 14 pmol/10(9) platelets) as compared to controls (59.0 +/- 4.3 pmol/10(9) platelets, p less than 0.01). Platelet exogenous sialytransferase activity and platelet aggregation were inhibited by aspirin. Thrombosis and bleeding have complicated the clinical course of half of these patients. This platelet membrane analysis provides additional data which may be related to current observations of increased levels of plasma sialytransferase activity and serum sialic acid in cancer patients.

Low erythrocyte pyridoxal-kinase activity in Blacks: Its possible relation to falciparum malaria.

The red-cell glucose-6-phosphate dehydrogenase (G.-6-P.D.) activity and red-cell pyridoxal-kinase (P.L.K.) activity of 27 Nigerian children with severe Plasmodium falciparum parasitaemia were compared with those of 26 healthy Nigerian children and 6 White adults. The mean P.L.K. activity of the malaria patients was similar to that of the Whites but significantly higher than that of the Nigerian controls. Correction for reduced mean red-cell age in patients was made by comparing the P.L.K.: G.-6-P.D. ratio for those subjects with stable G.-6-P.D. phenotypes. The mean P.L.K.:G.-6-P.D. ratio was the same for malaria patients and adult White but significantly higher than that for the Nigerian controls. These results suggest that the relatively high frequency of low red-cell P.L.K. activity among Blacks may have been selected for by falciparum malaria.

The depressant influence of extracellular fluid hyperoxia on liver slice oxygen uptake.

The effect of oxygen tension (Po2) on oxygen consumption (Vo2) of rabbit liver slices was investigated to determine the relationship between extracellular fluid Po2 and liver slice Vo2. Seventy rabbits, 5 kg each, were sacrificed with air embolism. Liver slices (0.3 gm) in Krebs-Ringer's-phosphate (KRP) dextrose were placed in a constantly agitated, 37 degrees C, closed cuvette, and Po2 was continuously monitored down to 10 torr. The system was reoxygenated and closed, and observations were repeated. Hemoglobin concentration was measured on the liver slice homogenate, and a P50 was measured on autologous blood. The presence of small amounts of hemoglobin in the supernatant was confirmed by electrophoretic, spectrophotometric, and morphologic studies. The pH decreased from mean 7.47 to mean 7.37 during each run. The resultant increase in P50 was recalculated and included in the determination of each deltaO2 content/min (Vo2). Vo2 at Po2 30 torr was greater than Vo2 at Po2 90 torr (p less than 0.0001 in each case). Vo2 at Po2 30 torr was not significantly different in the first vs. the second run (p greater than 0.85). The critical oxygen tension for hepatocyte respiration appears to be 30 torr.

Simultaneous spectrophotometry of Fe2+ and Cu2+ in serum denatured with guanidine hydrochloride.

We show how iron and copper ions may be simultaneously measured in 0.2 ml of serum after using guanidine hydrochloride to cause their release from protein. Sensitive reagents for iron (2,4,6-tripyridyl-s-triazine) and copper (disodium 2,9-dimethyl-4,7-diphenyl-1, 10-phenanthroline disulfonate) facilitate such measurement. We eliminated spectral interference between the two metal-ion complexes by selective use of pH. Results compare well with those obtained by accepted methods in which the concentrations of these ions are measured independently.

In vivo hepatic and intestinal toxicity of sodium cyanate in rats: cyanate-induced alterations in hepatic glycogen metabolism.

To determine the hepatic and intestinal toxicity of sodium cyanate, this compound was administered to rats by orogastric tube (PO) or intraperitoneal injection (IP). At low dosage (50 mg. per kilogram per day PO for 8 weeks), the animals showed no clinical effects other than mild lethargy. They had normal intestinal absorption studies, but demonstrated decreased liver G6PD activity and a slight increase in hepatic glycogen. At higher dose levels (200 mg. per kilogram per day PO for 10 days, 400 mg. per kilogram per day PO for 3 days, and 100 mg. per kilogram per day IP for 10 days), the animals became very lethargic and developed hind-limb paralysis; many animals died during the period of dosing. The severity and rate of onset of symptoms increased proportionally with the dose level. Liver sections from rats receiving these higher doses showed striking increases in glycogen deposition. Activities of hepatic enzymes involved in glycogen synthesis and degradation were measured in rats receiving 200 mg. per kilogram per day PO or 100 mg. per kilogram per day IP. Significant decreases were noted in the activities of glucose-6-phosphatase and G6PD in PO-dosed rats. The activities of phosphorylase, UDPG-pyrophosphorylase, glycogen synthetase, phosphoglucomutase, and debrancher did not differ from control rats. In IP-dosed rats, significant decreases were observed in the activities of glucose-6-phosphatase, G6PD, phosphorylase, and UDPG-pyrophosphorylase, but not in the other glycogen-related enzymes. Our data suggest that sodium cyanate affects several enzymes of hepatic glycogen metabolism but that the enzymes vary in their susceptibility (glucose-6-phosphatase and G6PD greater than phosphorylase and UDPG pyrophosphorylase.

Simulation of the pentose cycle in lactating rat mammary gland.

A computer model representing the pentose cycle, the tricarboxylic acid cycle and glycolysis in slices of lactating rat mammary glands has been constructed. This model is based primarily on the studies, with radioactive chemicals, of Abraham & Chaikoff (1959) [although some of the discrepant data of Katz & Wals (1972) could be accommodated by changing one enzyme activity]. Data obtained by using [1-(14)C]-, [6-(14)C]- and [3,4-(14)C]-glucose were simulated, as well as data obtained by using unlabelled glucose (for which some new experimental data are presented). Much past work on the pentose cycle has been mainly concerned with the division of glucose flow between the pentose cycle and glycolysis, and has relied on the assumption that the system is in steady state (both labelled and unlabelled). This assumption may not apply to lactating rat mammary glands, since the model shows that the percentage flow through the shunt progressively decreased for the first 2h of a 3h experiment, and we were unable to construct a completely steady-state model. The model allows examination of many quantitative features of the system, especially the amount of material passing through key enzymes, some of which appear to be regulated by NADP(+) concentrations as proposed by McLean (1960). Supplementary information for this paper has been deposited as Supplementary Publication SUP 50023 at the British Museum (Lending Division) (formerly the National Lending Library for Science and Technology), Boston Spa, Yorks. LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1973) 131, 5.