Erythrocyte sodium ion transport system in DOC-salt, Goldblatt, and spontaneously hypertensive rats. Altered erythrocyte Na+ transport has been observed in relation to the pathogenesis of essential hypertension. In the present study, intracellular Na+ and K+ levels, Na(+)-K+ pump activity, Na(+)-K+ cotransport, and Na+ passive permeability were measured in erythrocytes of DOC-salt hypertensive (DSH) rats, two-kidney, one clip Goldblatt hypertensive (2KH) rats, and spontaneously hypertensive rats (SHR). The results were as follows: 1. In comparison with the control groups, no change in the erythrocyte Na+ level was noted in the DSH and 2KH groups, whereas a significant increase was seen in the SHR group. 2. Although no change was noted in the erythrocyte K+ level in the 2KH and SHR groups when compared with the control groups, a significant decrease was seen in the DSH group. 3. Na(+)-K+ pump activity of erythrocytes was not changed in the DSH and 2KH groups when compared with the control group, but a significant increase was noted in the SHR group. 4. Na(+)-K+ cotransport of erythrocytes was not changed in any hypertensive rats when compared with the controls. 5. Na+ passive permeability in the erythrocyte membrane was not changed in the DSH and 2KH groups when compared with the control groups, but a significant increase was noted in the SHR group. These findings suggest that increased erythrocyte Na+ levels in SHR are due to increased Na+ passive permeability of the erythrocyte membrane, and increased Na(+)-K+ pump activity may be compensating for the increased intracellular Na+ concentration in erythrocytes. Furthermore, the increase in Na+ passive permeability observed in SHR might not result from hypertension itself but from abnormalities in the erythrocyte cell membrane, because no increase in Na+ passive permeability was noted in either DSH or 2KH rats.


Cytomegalovirus infections and toxoplasmosis in heart transplant recipients in Sweden. The morbidity of cytomegalovirus (CMV) infection and toxoplasmosis was evaluated in 75 heart transplant recipients. Among the 73 patients who survived more than one week after transplantation, 16 (22%) acquired primary CMV infection and 30 (41%) had evidence of secondary infection. All CMV seronegative recipients receiving hearts from seropositive donors developed CMV infection. The majority of infections (42/46) occurred during the first 4 months after transplantation. Overall, the incidence of symptomatic CMV disease was 44%. The infections were generally mild and only 1 death was attributed to primary CMV disease complicated by bacterial septicaemia and multiple organ failure. The severity of CMV disease was greatest among those with primary infection. There were 3 cases of toxoplasmosis. Two patients were toxoplasma seronegative before transplantation and developed clinical and serological signs of infection 2-3 months after transplantation despite receiving organs from seronegative donors. Of toxoplasma seronegative recipients receiving allografts from seropositive donors 3/4 were prophylactically treated with pyrimethamine for 6 weeks. None developed clinical or serological signs of toxoplasmosis while one patient who received trimethoprim-sulfamethoxazole had a subclinical infection.


Prevalence of antibodies against hepatitis B and C viruses among different groups of medical staff. The prevalence of antibodies to hepatitis C virus (anti-HCV) among the medical staff of a department of infectious diseases and a dialysis unit was studied (n = 80). Among medical staff from some selected emergency and operating units at elevated risk for blood exposure (n = 231) the prevalence of antibodies to both hepatitis B and C viruses was determined. Three out of 311 staff members tested (0.96%) were positive for anti-HCV, 1 from an emergency unit and 2 from surgery departments. 14/231 tested for anti-HBc (6.1%) were positive. None of these was positive for HBsAg. Thus, the prevalence of anti-HCV among medical staff frequently dealing with anti-HCV positive patients or at elevated risk for blood exposure was low and did not differ considerably from what has been found among healthy blood donors. The prevalence of markers for passed hepatitis B was only slightly augmented compared to what has been reported from the general population.


[Hepatitis A outbreak in a kindergarten]. In November/December 1990 eight out of 38 children of a kindergarten in Basle were involved in an outbreak of hepatitis A, and further cases occurred among family members and friends of the children. A questionnaire survey and a serologic analysis including all families associated with the kindergarten was undertaken to investigate the source, extent and spread of the hepatitis A. Among 147 persons examined, 20 clinical cases of hepatitis A were diagnosed (9 adults, 11 children), 5 more children were unapparently infected. The hepatitis A virus was imported by an adult who contracted the disease during a journey to Mexico. The virus spread from person to person, the third case being the kindergarten child, who was responsible for the actual outbreak. In a first phase, 11 persons were affected, 6 of them children from the kindergarten. The second phase involved 5 adults and another child from the kindergarten. The hygienic measures undertaken in the kindergarten proved to be efficacious; only one further child was infected thereafter. Immunoglobulin was protective if administered in time. Only 2 out of 21 passively immunized persons who had had contact with hepatitis A patients contracted an unapparent infection, but without further transmission. Among 82 persons who were passively immunized too late or not at all, 20 fell clinically ill with hepatitis A and 3 were unapparently infected. Use of the newly available hepatitis A vaccine is discussed.


Torque-velocity relation and muscle fibre characteristics of foot dorsiflexors after long-term overuse of residual muscle fibres due to prior polio or L5 root lesion. Isokinetic foot dorsiflexion strength and muscle biopsy data from eight patients with overuse of tibialis anterior muscle (TA) fibres due to weakness after prior polio or L5 root lesion were compared to data from age and sex matched, healthy subjects. Concentric peak torque at an angular velocity of 30 degrees/s was 6-24 Nm in the patients and 15-34 Nm in the controls (p < 0.01). Muscle biopsies from TA showed a significantly higher proportion of type 1 fibres in the patients as compared to the controls (p < 0.005). The type 1 fibres in the patients had a significantly larger cross-sectional area than in the controls (p < 0.005). The type 1 fibre proportion and relative area were inversely correlated to the relative concentric torque produced at 180 degrees/s (p < 0.05) and 240 degrees/s (p < 0.05) compared to that produced at 30 degrees/s in the controls but not in the patients. However, the relative concentric torque produced at 180 degrees/s and 240 degrees/s compared to that produced at 30 degrees/s was not significantly different in the patients and the controls. This indicates that the contractile properties of the overused muscle fibres do not change in parallel with the histochemical fibre type.


[Stress-induced carpal tunnel syndrome in athletes--exemplified by 3 kinds of sports]. As is generally known in occupational medicine, exposure to repetitive wrist movements, to vibration and heavy manual work may cause a carpal tunnel-syndrome (CTS). The examination of three kinds of sport (swimming, moto-cross-riding, body-building), each typical for one of the described reasons of high risk for an exercise-induced CTS, showed on 18 sports-women and 54 sportsmen that prolonged practice of one of these kinds of sport may indeed lead to CTS among young people. Six of the examined persons on the typical symptoms of CTS. The electrophysiological main feature of CTS, namely the distal motor latency of the median nerve, was had nine other sportsmen significantly longer than the age-specific normal value (according to Ludin). A clear positive correlation between the estimated total duration of training and the distal motor latency of the median nerve was found.


The cytosensor microphysiometer: biological applications of silicon technology. A silicon-based device, dubbed a microphysiometer, can be used to detect and monitor the response of cells to a variety of chemical substances, especially ligands for specific plasma membrane receptors. The microphysiometer measures the rate of proton excretion from 10(4) to 10(6) cells. This article gives an overview of experiments currently being carried out with this instrument with emphasis on receptors with seven transmembrane helices and tyrosine kinase receptors. As a scientific instrument, the microphysiometer can be thought of as serving two distinct functions. In terms of detecting specific molecules, selected biological cells in this instrument serve as detectors and amplifiers. The microphysiometer can also investigate cell function and biochemistry. A major application of this instrument may prove to be screening for new receptor ligands. In this respect, the microphysiometer appears to offer significant advantages over other techniques.


Ultrafast infrared spectroscopy.

Ultrafast infrared spectroscopy. Recent advances in ultrafast infrared spectroscopy are described, including experimental details and fundamental limitations. The utility of this technique is illustrated with two recent examples.


Association of human cyclin E with a periodic G1-S phase protein kinase. G1 cyclins control the G1 to S phase transition in the budding yeast, Saccharomyces cerevisiae. Cyclin E was discovered in the course of a screen for human complementary DNAs that rescue a deficiency of G1 cyclin function in budding yeast. The amounts of both the cyclin E protein and an associated protein kinase activity fluctuated periodically through the human cell cycle; both were maximal in late G1 and early S phases. Cyclin E-associated kinase activity was correlated with the appearance of complexes containing cyclin E and the cyclin-dependent kinase Cdk2. Thus, the cyclin E-Cdk2 complex may constitute a human G1-S phase-specific regulatory protein kinase.


Molecular diversity of glutamate receptors and implications for brain function. The glutamate receptors mediate excitatory neurotransmission in the brain and are important in memory acquisition, learning, and some neurodegenerative disorders. This receptor family is classified in three groups: the N-methyl-D-aspartate (NMDA), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)-kainate, and metabotropic receptors. Recent molecular studies have shown that many receptor subtypes exist in all three groups of the receptors and exhibit heterogeneity in function and expression patterns. This article reviews the molecular and functional diversity of the glutamate receptors and discusses their implications for integrative brain function.


Flow-dependent cytosolic acidification of vascular endothelial cells. Hemodynamic shear stress affects endothelial cell structure and function, but little is known about the signal transduction mechanisms involved in these processes. The effect of laminar shear stress on cytosolic pH (pHi) was examined in rat aortic endothelial cells cultured in glass capillary tubes. Shear stress forces led to a rapid decrease in pHi (maximal effect 0.09 pH unit at 13.4 dynes per square centimeter). Removal of specific ions or addition of exchange inhibitors suggests that in vascular endothelial cells shear stress forces activate both an alkali extruder, sodium ion-independent chloride-bicarbonate ion exchange, and an acid extruder, sodium-hydrogen ion exchange; the net effect in physiologic buffer with the bicarbonate ion is a decrease in pHi.


Role of transmembrane domain interactions in the assembly of class II MHC molecules. Evidence is presented that suggests a role for transmembrane domain interactions in the assembly of class II major histocompatibility complex (MHC) molecules. Mutations in the transmembrane domains of the class II MHC alpha or beta chains resulted in proteins that did not generate complexes recognized by conformation-dependent antibodies and that were largely retained in the endoplasmic reticulum. Insertion of the alpha and beta transmembrane domains into other proteins allowed the chimeric proteins to assemble, suggesting a direct interaction of the alpha and beta transmembrane domains. The interactions were mediated by a structural motif involving several glycine residues on the same face of a putative alpha helix.


Burst firing in dopamine neurons induced by N-methyl-D-aspartate: role of electrogenic sodium pump. Dopamine-containing neurons of the mammalian midbrain are required for normal behavior and movements. In vivo they fire action potentials in bursts, but in vitro they discharge regularly spaced action potentials. Burst firing in vitro has now been shown to be robustly induced by the glutamate agonist N-methyl-D-aspartate (NMDA) although not by the non-NMDA agonists kainate or quisqualate. The hyperpolarization between bursts of action potentials results from electrogenic sodium ion extrusion by a ouabain-sensitive pump. This mechanism of burst generation in mammalian neurons may be important in the pathophysiology of schizophrenia and Parkinson's disease.


Clinical trials with ifosfamide: the Indiana University experience. At Indiana University, we began clinical trials with ifosfamide in 1981. Although our initial efforts were in a variety of tumor types, including pancreatic cancer, we have most recently focused our attention on two tumors that have historically exhibited a higher degree of chemosensitivity--testicular cancer and small cell lung cancer (SCLC). In phase II trials, ifosfamide has proven to have single-agent activity in both diseases. Coupling this data with preclinical observations of synergy with cisplatin and etoposide, we began trials of ifosfamide plus cisplatin with either vinblastine (VeIP) or etoposide (VIP) in patients with recurrent germ cell tumors; we also investigated the use of VIP in SCLC. In third-line or greater therapy for recurrent germ cell tumors, a 36% disease-free status was attained, with 16% of patients continuously free of disease for 5 or more years. Currently, ifosfamide is being evaluated as part of initial therapy in patients with advanced disease. In SCLC, VIP has also been investigated as part of initial therapy in patients with extensive disease. The complete response rate of 38% achieved in 37 evaluable patients has spurred the Hoosier Oncology Group to compare the VIP regimen to cisplatin/etoposide in patients with extensive-disease SCLC. Ifosfamide has the broad range of clinical activity, but its ultimate role as part of initial therapy remains to be discerned.


Ifosfamide in non-small cell lung cancer: a review. In recent years, ifosfamide has proven to be one of the most effective cytostatic drugs in the treatment of non-small cell lung cancer. Single-agent studies have demonstrated response rates of 15% to 30%. Combinations containing ifosfamide together with one or two other cytostatic agents yield overall response rates of 30% to 45%, with 5% to 10% complete remissions. For this purpose, ifosfamide has proven to be an ideal constituent, especially since the routine use of mesna uroprotection has significantly reduced its toxicity. As myelosuppression is now the primary dose-limiting toxicity, further investigations are directed toward the question of whether adjuvant administration of hematopoietic growth factors can improve treatment results by allowing increased dose intensity. This seems to be particularly important in patients with malignancies that respond poorly to cytostatic chemotherapy to determine whether survival can be significantly prolonged. Further studies with ifosfamide, moreover, should include neoadjuvant investigations in patients with stages II and III disease or its combination with other treatment modalities.


Phase II study of cisplatin, ifosfamide with mesna, and etoposide (PIE) chemotherapy for advanced non-small cell lung cancer. Ifosfamide has shown promising single-agent activity in non-small cell lung cancer (NSCLC). We combined ifosfamide (1,800 mg/m2 plus mesna 1,100 mg/m2 by intravenous [IV] continuous infusion daily for 3 days) with cisplatin (20 mg/m2 IV for 3 days) and etoposide (80 mg/m2 IV for 3 days) and treated 41 chemotherapy-naive patients with recurrent or metastatic NSCLC. Fifteen (40.5%) of the 37 evaluable patients had objective responses (1 complete and 14 partial). Patients with good performance status (Zubrod scale 0 or 1) had a higher response rate than the patients with poor performance status (Zubrod scale 2) (11 of 21 patients [52.4%] versus four of 16 [25.0%]), but the difference was not statistically significant (P = .09). Median survival has not been reached, with 17 patients still alive after a median follow-up of 39 weeks (range, 21 to 56). Significant myelosuppression occurred, with granulocytopenia being the dose-limiting toxicity. Overall, treatment was well tolerated. Considering the pooled response rate of 32% with cisplatin/etoposide regimens and the previous experience from our institution, the results with this three-drug regimen are very encouraging, and its further investigation is warranted.


Ifosfamide, cisplatin, and etoposide (ICE) in the treatment of advanced non-small cell lung cancer. Forty-seven previously untreated patients with histologically or cytologically proven non-small cell lung cancer were treated with ICE (ifosfamide/cisplatin/etoposide). Patients received ifosfamide 4 g/m2 with mesna uroprotection on day 1, and cisplatin 25 mg/m2/d and etoposide 100 mg/m2/d on days 1, 2, and 3; courses were repeated every 28 days. Premedication with prochlorperazine, dexamethasone, and high-dose metoclopramide was given to prevent nausea; lorazepam was added on days 2 and 3 only. Thirty-four men and 13 women (median age, 60 years) received a total of 146 treatment cycles. One patient had stage IIIA disease, seven had IIIB disease, and 39 had hematogenous metastases. Forty-six patients were evaluable for response and toxicity. One patient suffered a myocardial infarction on day 7 that was judged unrelated to treatment. Two patients suffered early death from toxicity and have been classified as nonresponders. Three patients achieved complete response (median, 42+ weeks) and 14 patients achieved partial response (median, 29+ weeks; range, 10 to 82+), for an overall response rate of 37% (95% confidence limits, 23% to 51%). The median survival of the entire group is 26 weeks (1 to 82+). The median nadir granulocyte count was 0.275 x 10(9)/L (range, 0 to 2.3 x 10(9)/L), and there were 14 episodes (in 11 patients) or neutropenia-associated fever, one of which resulted in death. Seven of these patients had not had the required protocol dose reduction for nadir neutrophil count in the preceding cycle. The median nadir platelet count was 120 x 10(9)/L (range, 13 to 385 x 10(9)/L), and three patients required platelet transfusions. Eleven patients had RBC transfusions. Only ten patients had grade 2 gastrointestinal toxicity. Five patients had microscopic hematuria, and one patient had central nervous system toxicity.


Overview of ifosfamide in small cell lung cancer. Ifosfamide, an analogue of the alkylating agent cyclophosphamide, is one of the most active agents in the treatment of small cell lung cancer (SCLC). As a single agent, the drug produces a greater than 50% objective response rate. Recent studies using ifosfamide in combination with other active agents, ie, cisplatin, carboplatin, and etoposide, In the treatment of limited-stage and extensive-stage SCLC have achieved high overall response rates and complete responses (CRs). However, the CR rate is higher in limited-disease patients. Additional studies are needed with the various ifosfamide-containing regimens to precisely define their role in the treatment of SCLC in both limited-disease and extensive-disease patients.


Novel approaches with ifosfamide in small cell lung cancer. During the past 5 years, ifosfamide has been used increasingly in combination chemotherapy for small cell lung cancer (SCLC). The high activity and favorable toxicity spectrum (with the uroprotector mesna) will encourage further use. A policy of no dosage reduction is feasible in patients receiving combination chemotherapy with ifosfamide given as a 24-hour intravenous (IV) infusion, which is much more convenient than the more commonly used 4- to 5-day fractionated regimen. This policy has resulted in actual 2-year survivals of 22% to 33% among SCLC patients not intensively staged. The stability of ifosfamide and its high bioavailability have allowed its use in chronic, 7-day ambulatory IV infusions, with decreased toxicity and hospitalization. Recently, oral and subcutaneous administration also have been tried, again allowing outpatient treatment. The first studies with hemopoietic growth factor support, eg, granulocyte colony-stimulating factor, conducted with combination chemotherapy with ifosfamide-containing regimens in SCLC, demonstrated significant reduction in neutropenia, infections, and antibiotic use. It is clear that the dosage of ifosfamide can be intensified in the future. The broad versatility of the drug will allow interesting new studies, including those to be conducted with outpatients.


Clinical experiences with carboplatin (paraplatin) in lung cancer. Lung cancer is the most lethal cancer in the United States, with 143,000 deaths predicted for 1991. The cure rate is extremely low (approximately 13%), in part because the propensity for early spread precludes surgical cure in most patients. Thus, chemotherapy or other systemic therapies are the only way to improve the dismal results. Cisplatin is an active agent in small cell lung cancer (SCLC) and perhaps the most active agent in nonsmall cell lung cancer (NSCLC). The toxicities and inconvenience of cisplatin make it less than ideal for lung cancer therapy. Carboplatin was developed to provide a less toxic, more convenient alternative to cisplatin. The data presented in this review suggest that carboplatin may be substituted for cisplatin in the treatment of extensive-stage SCLC. In limited-stage SCLC, there are insufficient data to determine whether it should replace cisplatin when used simultaneously with chest irradiation and etoposide. It may be substituted for cisplatin in cycles not using irradiation. In NSCLC, carboplatin may be used alone or with etoposide for the palliative management of metastatic disease. Its role in earlier stages of NSCLC needs investigation.


Carboplatin-containing regimens for small cell lung cancer: implications for management in the elderly. The efficacy and toxicity of two regimens based on etoposide/carboplatin with or without cyclophosphamide/vincristine in the management of small cell lung cancer (SCLC) were assessed by the Australian Lung Cancer Study Group. Response rates of 77% and 85% were noted for the two- and four-drug regimens, respectively, among patients with limited disease (LD). Response rates among patients with extensive disease (ED) were 58% and 79%, respectively. The profiles of nonhematologic toxicity were modest; myelosuppression was dose-limiting when colony-stimulating factors were not used. Twenty-six patients (14%) were older than 70 years of age. Although hematologic toxicity was more severe in the elderly group, there was no significant difference in nonhematologic toxicity, response rate, or overall survival between the geriatric and younger groups. When LD only was considered, 33% of those younger than 70 were alive at 2 years; no patients aged 70 years or older with LD were alive beyond 2 years. In patients with ED, there was no age-related difference in survival. Cytotoxic regimens based on etoposide/carboplatin constitute useful treatment for SCLC, with high response rates and manageable toxicity, irrespective of patient age.


Dose-intensive therapy for germ cell neoplasms. Dose-intensive chemotherapy for recurrent or refractory germ cell cancer has evolved over the last decade. Initial experience using high-dose regimens including single agents like cyclophosphamide, thiotepa, and etoposide demonstrated that responses could be obtained in the majority of patients but that they tended to be of short duration. Second-generation studies focusing on this disease site incorporated drugs and principles specific to germ cell cancer. Large studies performed at Indiana University (Indianapolis, IN) and in several European centers have demonstrated that the addition of high-dose carboplatin or cisplatin to other regimens can result in long-term survival of otherwise incurable patients. With the results of initial pilot studies now confirmed, high-dose carboplatin-based chemotherapy is beginning to play a role in first salvage therapy, and in some centers is being used in protocols of initial therapy for poor-risk patients. Based on the success of some of the early studies in germ cell cancer, similar protocols also are being investigated for other disease sites such as ovarian cancer, neuroblastoma, lymphoma, and breast cancer.


Second-line carboplatin-based chemotherapy for small cell lung cancer: the Groningen experience. In this report, the results of two phase II studies and one pilot study of second-line carboplatin-based chemotherapy for small cell lung cancer are described. Carboplatin plus vincristine given with or without ifosfamide resulted in response rates of 36% and 53%, respectively, in so-called chemotherapy-resistant patients. Toxicity of the carboplatin/vincristine regimen was mild (hematologic toxicity grade 4 was seen with 13% of the courses), whereas the combination including ifosfamide resulted in grade 4 thrombocytopenia in 57% of the courses and grade 4 leukocytopenia in 49%. A partial response was seen in one of nine patients with progression of brain metastases after chemotherapy, and in three patients the neurologic function score improved, with minor tumor reduction evident on computed tomography scan of the brain. We conclude that carboplatin is a useful agent for second-line chemotherapy in patients with an early relapse after induction chemotherapy.


Phase II and III studies with carboplatin in small cell lung cancer. Carboplatin is one of the most active agents in untreated small cell lung cancer (SCLC) (11% complete response [CR], 59% CR plus partial response [PR]). Combination carboplatin/etoposide/vincristine (CEV) (phase II trial) led to an overall remission rate of 84% in patients with limited disease (LD), with 52% CRs. The median survival time with this combination was 14 months in patients with LD and 9.5 months in those with extensive disease (ED). The 30-month survival rates are 30% in LD and 10% in ED, which represents a plateau of the survival curve. This regimen is highly effective and exhibits low toxicity in SCLC. To evaluate the role of carboplatin in combination chemotherapy in patients with extensive SCLC, a phase III trial was performed. In this ongoing trial comparing CEV and etoposide/vincristine in ED patients, CR and overall response rates to date are higher in the CEV arm (CR, 20% versus 15%; CR plus PR, 83% versus 65%), but as yet the differences are not statistically significant. In summary, chemotherapy regimens containing platinum compounds are among the most active in the treatment of SCLC. The use of the new compound carboplatin instead of cisplatin has led to similar or increased remission rates and is preferable because it has fewer side effects. Preliminary results from this ongoing, prospective, randomized phase III trial are presented.


Serum neuron-specific enolase and lactate dehydrogenase as predictors of response to chemotherapy and survival in non-small cell lung cancer. The prognostic value of serum neuron-specific enolase (NSE) and lactate dehydrogenase (LDH) was prospectively assessed in 42 patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) treated within two chemotherapy trials. Pretreatment NSE levels ranged from 4.6 to 34.6 ng/mL (median value, 7.5) and were above 10 ng/mL in ten patients (23.8%). LDH levels varied between 85 U/L and 2,484 U/L (median value, 220) and were above 250 U/L in 12 patients (29.3%). Elevated levels of both enzymes were significantly more common in patients with metastatic disease than in those with locoregional disease (40% v 9% for NSE and 40% v 18% for LDH, respectively). Strong positive correlation (correlation factor 0.693) was found between NSE and LDH serum levels. The levels of both markers did not correlate with age, sex, previous therapy, performance status, or histology. Responses to chemotherapy were seen more frequently in patients with elevated NSE levels (six of ten, 60%) than in those with normal values (seven of 32, 22%; P = .02). Similar correlation was found for LDH: Response was seen in seven of 12 patients with elevated levels (58%) and in six of 29 (21%) of those with normal values (P = .02). In a logistic regression analysis, both markers considered individually remained significant when adjusted by sex, age, performance status, prior therapy, histology, and extent of disease. Three pretreatment characteristics, high levels of NSE and LDH (both considered as continuous variables) and metastatic disease, were found to be associated with shorter survival; with adjustment for extent of disease, however, NSE and LDH were no longer correlated with shorter survival. These data suggest potential clinical value of NSE and LDH determination in treatment selection of NSCLC patients.


Carboplatin in non-small cell lung cancer: an update on the Cancer and Leukemia Group B experience. Since 1984, the Respiratory Committee of the Cancer and Leukemia Group B (CALGB) has evaluated carboplatin, either alone or in combination, in five separate phase II studies for patients with inoperable non-small cell lung cancer (NSCLC). All patients had an Eastern Cooperative Oncology Group performance status of 0 to 2 and had not received previous treatment with chemotherapy. In 70 patients with stage IIIB or IV disease, carboplatin 400 mg/m2 administered intravenously once every 4 weeks produced a 16% overall response rate and an acceptable toxicity profile. Subsequently, combinations of carboplatin/cisplatin, carboplatin/etoposide, and carboplatin/vinblastine have been evaluated in similar patient groups. Response rates of 11%, 12%, and 20%, respectively, were obtained. Myelosuppressive toxicity was substantially greater with carboplatin/etoposide and carboplatin/vinblastine than with carboplatin alone. Carboplatin/vinblastine demonstrated efficacy similar to that of the cisplatin/vinblastine combination previously evaluated by CALGB for treatment of similar patients with advanced NSCLC; ease of administration and lack of significant nephrotoxicity also compared favorably with cisplatin-based therapy. In regional NSCLC patients, carboplatin 100 mg/m2/wk can be administered intravenously concurrently with 60 Gy thoracic radiotherapy given over 6 weeks. The impact of concurrent carboplatin added to a sequential chemotherapy-radiotherapy program for patients with regional NSCLC is currently under study by the CALGB Respiratory Committee.


Carboplatin plus oral etoposide in the management of advanced, non-small cell lung cancer: preliminary results of a Vanderbilt trial. Twenty-eight patients with unresectable, metastatic non-small cell lung cancer (NSCLC) were treated with carboplatin/oral etoposide. Carboplatin was administered intravenously on day 1 at a dose of 300 mg/m2 (12 patients) or 350 mg/m2 (16 patients); oral etoposide was administered at a dose of 50 mg/m2/d for 21 consecutive days. Treatment was repeated every 28 days. Patient characteristics included male:female ratio of 23:5, median age of 60 years, median Eastern Cooperative Oncology Group performance status of 1, weight loss of 5% or more in seven patients; stage IIIB disease in two patients and stage IV in 26. Twenty-five patients were evaluable for response and seven (28%) achieved a partial response (95% confidence interval, 14% to 48%). Median duration of response was 3+ months (range, 2+ to 6+) and median survival was 4+ months (range, 1+ to 10+). Myelosuppression was the predominate toxicity; leukocyte and platelet nadirs occurred between days 22 and 29, with median counts of 2,900/microL and 172,000/microL, respectively. The median interval between the start of cycle 1 and the start of cycle 2 was 33 days (range, 26 to 42). Carboplatin/oral etoposide is a moderately active regimen against advanced NSCLC that can be administered in an outpatient setting with manageable toxicity. Its impact on survival remains to be determined.


Use of carboplatin in germ cell tumors of the testis. The high cure rate among young men with metastatic testicular germ cell tumors heightens the issue of late treatment toxicity. Conventional combination chemotherapy based on cisplatin can be expected to cure 85% to 90% of patients with metastatic disease; however the cost of this success includes severe treatment side effects, including irreversible renal damage and the risk of neurotoxicity and ototoxicity. Carboplatin was investigated as a replacement for cisplatin in the treatment of these tumors because of its reduced toxicity. The Royal Marsden Hospital pilot studies have included 76 patients treated for metastatic nonseminoma with the combination carboplatin/etoposide/bleomycin between 1984 and 1988. The median follow-up at time of analysis was 24 months. The complete remission rate was 95% and the overall cause-specific survival was 98.5%. Also, 33 patients treated with single-agent carboplatin for advanced metastatic seminoma were followed for a median of 36 months. The actuarial progression-free survival was 79%, and 91% of patients were alive and disease-free. The results of these studies indicate that carboplatin has activity equivalent to cisplatin in germ cell tumors of the testis and is less toxic.


Mechanisms of action of teniposide (VM-26) and comparison with etoposide (VP-16). Teniposide is the result of extensive, long-term efforts to refine and improve on the cytotoxic activity of naturally occurring compounds extracted from podophyllin resins and purified. Isolation of an extremely potent though minor component of one of the early podophyllin derivatives led in turn to the synthesis and evaluation of several aldehyde condensation products. Two of these, teniposide and etoposide, were further investigated when their considerable antitumor activity in animals became apparent. Recognition of transient DNA breaks induced by teniposide, etoposide, and other podophyllotoxin analogues established not only that their site of activity was DNA but also that their cytotoxic effect was dose-dependent. Extensive investigation has further indicated that a primary mechanism of action of these agents involves inhibition of the catalytic activity of eukaryote topoisomerase II and, more important, the consequent stabilization of the normally transient covalent intermediate formed between the DNA substrate and the enzyme. As a result of elevated enzyme levels or enzyme activity, or both, in transformed cells, topoisomerase II inhibitors are highly selective for cancer cells versus normal cells. Although teniposide is not substantially more potent than etoposide in terms of catalytic inhibition or stabilization of the DNA-enzyme intermediate, it is more readily taken up by cells, which results in greater teniposide accumulation within the cells and, thus, a greater capacity for cytotoxicity.


Phase I study of oral teniposide (VM-26). A phase I study of teniposide administered orally for 5 consecutive days was performed. The first dose was 60 mg/m2/d, with increments of 25 mg/m2/d. Nineteen patients entered the study and received a total of 77 courses with a median of two (range, 1 to 12). Dose-limiting toxicity occurred at 160 mg/m2/d and consisted of myelosuppression, mainly leukocytopenia, and gastrointestinal toxicity. Sufficient recovery of blood counts was seen by day 21 to allow for a repeat course. Two patients, treated with teniposide doses of 110 and 160 mg/m2/d, respectively, were considered toxic deaths. Partial alopecia was frequent at doses above 85 mg/m2/d. Retching and vomiting during administration of the drug were encountered in virtually all patients. Oral teniposide 135 mg/m2/d for 5 consecutive days on a 3-week schedule is recommended for evaluation of antitumor efficacy in phase II studies.


Teniposide in the treatment of small cell lung cancer: a review. The epipodophyllotoxin derivatives teniposide and etoposide have been under clinical investigation for over 15 years. Although etoposide has been established as one of the most active compounds in the treatment of small cell lung cancer (SCLC), teniposide has received little attention. The results of seven phase II studies evaluating response rate and duration of response to teniposide in 10 previously treated and 102 untreated patients showed response rates of 21% and 58%, respectively. The most frequently used dosage schedule was 60 mg/m2 intravenously daily for 5 days every 3 weeks. The following are factors influencing the response rate and duration of response to teniposide: performance status; prior weight loss; prior chemotherapy exposure, including prior treatment with etoposide; stage; and effectiveness of prior chemotherapy, including time from last administration. Preliminary analyses from a study comparing the efficacy of teniposide with that of etoposide suggest that teniposide may be more effective in previously untreated patients with SCLC who are 70 years of age or older. The preliminary data, however, indicate that equivalent doses of teniposide cause more cases of leukopenia than etoposide. Before a final conclusion can be drawn, the results from an ongoing study using teniposide and etoposide at equitoxic doses must be evaluated.


Teniposide (VM-26) and carboplatin as initial therapy for small cell lung cancer. Forty-four patients with previously untreated histologically proven small cell lung cancer (SCLC) were treated with a combination of teniposide 60 mg/m2 intravenously (IV) on days 1 through 5 and carboplatin 400 mg/m2 IV on day 1 every 28 days for six courses. Patients with limited disease (LD) subsequently received prophylactic cranial and thoracic radiotherapy. Of the 44 patients, 40 were evaluable for response: 31 (78%) achieved an objective response; 9 of 18 patients (50%) with LD had a complete response (CR), with a partial response (PR) plus CR rate of 78%. Two of 22 patients (9%) with extensive disease achieved a CR, with a combined PR and CR rate of 77%. Median duration of response for all evaluable patients was 253 days (36 weeks). Median duration of survival for LD patients was 368 days (52 weeks). Survival of LD patients was 86% at 6 months, 52% at 12 months, and 26% at 18 months. Median duration of survival for all patients in the study was 275 days, with a survival of 79% at 6 months, 36% at 1 year, and 12% at 18 months. Myelosuppression was the main toxicity, with World Health Organization (WHO) grade 3 or 4 infection occurring in 38% of patients. However, no patient died of sepsis or hemorrhage. Treatment was otherwise well tolerated, with no neurotoxicity or nephrotoxicity documented. The high activity of this drug combination justifies its use as first-line treatment of previously untreated SCLC.


Teniposide alone and in combination chemotherapy in small cell lung cancer. Teniposide is one of the most active agents against small cell lung cancer (SCLC). In a phase II study, teniposide achieved a 90% response rate in 33 untreated elderly patients. At our institution, teniposide produced a 34% response rate in a group of 44 unselected patients. Pilot studies of combination chemotherapy with teniposide have recently been initiated. A phase II trial with teniposide, vincristine, methotrexate, and cyclophosphamide in SCLC patients was started, based on demonstration of experimental synergy between these drugs. Chest irradiation was also given to patients with limited disease who responded to chemotherapy, and prophylactic cranial irradiation was given to complete responders (CRs). A response rate of 78% with 22% CRs was achieved in 32 evaluable previously untreated SCLC patients; median durations of response and survival were 252 and 311 days, respectively. Main side effects were myelosuppression, mucositis, and peripheral neuropathy. This teniposide combination chemotherapy compares favorably with other reported active regimens in SCLC. Further trials will determine whether the introduction of teniposide in combination chemotherapy is able to improve the outcome of SCLC.


Treatment of brain metastases of small cell lung cancer with teniposide. Over 50% of patients with small cell lung cancer (SCLC) will develop symptomatic brain metastases during the course of their disease. Results of whole brain radiotherapy, the standard treatment, are rather poor and relapses are frequent. Thus, new modes of therapy are urgently needed for these patients. In this study, the efficacy of teniposide was evaluated at a dose of 150 mg/m2 intravenously on days 1, 3, and 5 at 3-week intervals. In 11 of 26 evaluable patients an intracranial response was observed. Median response duration was 23 weeks (range, 9 to 50). Toxicity was acceptable, with grades 3/4 leukocytopenia and thrombocytopenia reported in 37% and 16%, respectively, of 123 courses. Therefore, teniposide is an effective agent against brain metastases of SCLC and is suitable for palliation of these patients.


The European Organization for Research and Treatment of Cancer experience with teniposide: preliminary results of a randomized study in non-small cell lung cancer. Chemotherapy for non-small cell lung cancer (NSCLC) is unsatisfactory, and the search for new active drugs has been relatively unsuccessful. Most polychemotherapy regimens in NSCLC include cisplatin with a vinca alkaloid or etoposide. Among the new agents tested in recent years, teniposide produced a 17% response rate in 42 evaluable patients, with a 21% response rate in untreated patients. The Lung Cancer Cooperative Group of the European Organization for Research and Treatment of Cancer has started a randomized trial comparing two different schedules of teniposide administration with and without cisplatin. This paper reports the preliminary findings for the initial 80 patients in this randomized study.


Breast carcinoma-in-situ: an emerging problem in Singapore. Breast carcinoma-in-situ constitutes 4.1% of 707 breast cancers diagnosed between 1988 and 1990. Among these 30 patients, intraductal carcinoma-in-situ (DCIS) outnumbers lobular carcinoma-in-situ (LCIS) by 9-fold. They are mostly symptomatic - 87% present as breast lumps and/or nipple discharge, with 52% of lumps exceeding 2cm size. Three patients were detected by screening mammography and it is expected that more breast carcinoma-in-situ will be detected through mammographic screening. Two-thirds of the patients had mastectomy while the rest had lesser procedures. The different surgical procedures and adjuvant therapy instituted for the patients are reflections of the differing opinions regarding optimum therapy for carcinoma-in-situ and the differing rationale for DCIS and LCIS lesions.


Paraneoplastic limbic encephalopathy as a nonmetastatic complication of small cell lung cancer. A 52-year-old man who presented symptoms compatible with paraneoplastic limbic encephalopathy (PLE) was found to have small cell lung cancer. Antineoplastic therapy resulted in complete remission. Because the neuropsychiatric symptoms are potentially reversible, it is important to recognize these symptoms as a manifestation of malignancy so that treatment can be instituted.


Disseminated strongyloidiasis in a World War II Veteran with metastatic undifferentiated carcinoma of neuroendocrine type. This case of disseminated strongyloidiasis illustrates the need for a high index of suspicion and effective screening for this pathogen in certain high-risk patient populations before giving immunosuppressive therapy. A complete travel and military history should be obtained. In the United States, persons with a history of military service in endemic areas, emigrants from endemic areas, patients in institutions, and residents of the Southeast should have careful evaluation for strongyloidiasis. Eosinophilia may or may not be present. Stool examinations are not sufficient to rule out the diagnosis. When available, the ELISA screening test for serodiagnosis may be useful. Duodenal aspirates or bronchoalveolar lavage may be necessary to confirm the diagnosis.


Human papillomavirus DNA detection in primary anogenital warts and cervical low-grade intraepithelial neoplasias in adults by in situ hybridization. In this study, 58 consecutive patients with primary anogenital warts were selected from patients attending a genitourinary clinic. Patients were grouped on the basis of clinical lesion site, i.e. patients with genital warts only, patients with perianal or anal canal warts only, and patients with concurrent perianal and genital warts. Of these patients, 38% of the men (12/31) and 33.3% of the women (9/27) had other anogenital infections, such as nonspecific urethritis (NSU) or nonspecific genital infection, which were the most common. Of the patients who had perianal warts, 37% of the men (7/19) and 25% of the women (4/16) also had warts in the anal canal. Of the women who had anogenital warts, 63% (17/27) had concurrent subclinical low-grade cervical intraepithelial neoplasia (CIN) lesions. Human papilloma virus (HPV) DNA (either 6 or 11, 16 or 18, or 31 or 33 or 35) was detected in 53.3% (40/75) of the anogenital wart biopsy samples, and in 35.2% (6/17) of the low-grade CIN lesions. HPV types 6 or 11 were the most common types in anogenital warts (45.3%); and in CIN lesions HPV types 6 or 11 and 16 or 18 were found with equal frequency (17.6% each). There were no significant differences in HPV types between patients with genital warts and patients with perianal and anal canal warts. Anogenital infection with HPV is multicentric; external anogenital warts and subclinical CIN lesions often exist concurrently. The low prevalence of HPV DNA detected in anogenital warts and CIN biopsy samples may be due to insensitivity of the in situ hybridization technique used in this study.


Results of hepatic resection and transplantation for fibrolamellar carcinoma. Fibrolamellar carcinoma (FLC) is a tumor of the liver that can be differentiated from common hepatocellular carcinoma (HCC). Despite the exceptional role of the clinicopathologic signs and symptoms, true appraisal of the prognosis of the tumor is not clear and remains a controversial issue. To determine the long term prognosis of FLC more precisely, a retrospective study of 20 consecutive patients was performed, with analysis of selected pathologic factors, particularly the TNM staging system. Curative tumor removal (R0) was achieved by partial hepatic resection in 14 patients and total hepatectomy with subsequent replacement of the liver in six patients, respectively. The estimated overall five year survival rate was 36.6 percent. There was an advantage of partial versus total hepatectomy, with median survival times of 44.5 versus 28.5 months. Statistically significant better survival rates at five years were observed in patients with solitary tumors and in instances of absent regional lymph node metastases. Although other factors analyzed did not show significant differences, there was a tendency indicating individual tumor stage was the most significant determinant for prognosis. For further discussion of an apparently more favorable outcome of patients with FLC as compared with common HCC, detailed specification of the tumor stages seems mandatory. From the present analysis, the fibrolamellar variant could not be confirmed to be an independent indicator of better patient survival. The treatment of choice remains radical operation. The goal can, at best, be achieved by a therapeutic concept including partial as well as total hepatectomy, depending on the stage of the tumor.


A prospective evaluation of distal margins in carcinoma of the rectum. The controversy regarding the ideal distal margin after a "curative" anterior resection is currently unresolved. To clarify this issue, a prospective study was undertaken. Two hundred and forty-three patients who underwent "curative" anterior resection were included in this study. Patients were divided into 1 centimeter groups based upon the length of the distal margin (from 0.0 to 8.0 centimeters), and recurrence, local and distant, and survival were analyzed. There was no significant difference in local or distant recurrence or survival when each centimeter interval was studied to 1 centimeter. Patients with a distal margin of less than 0.8 centimeter (group 1) had more frequent anastomotic recurrences (30 percent) when compared with patients (group 2) with a distal margin greater than 0.8 centimeter (10.5 percent; p = 0.01). Both groups were well matched for age, gender, tumor size, length of proximal margin, number of positive lymph nodes, histologic factors, grade of differentiation, Dukes' classification, type of operation, preoperative and postoperative radiotherapy, follow-up evaluation and surgeon. Both groups had a similar pelvic area recurrence (5.0 versus 8.7 percent, p = NS) and distance recurrence rates (10.0 versus 14.2 percent, p = NS). The five year survival rate was adversely affected by a distal margin of less than 0.8 centimeter (49.3 percent in group 1 and 67.5 percent in group 2; p = 0.01). The data suggest that a distal margin of 1 centimeter in the pathologic specimen is adequate distal clearance for most carcinomas of the rectum.


[Changes in cellular ion transport and arterial pressure in renal transplant recipients undergoing cyclosporin treatment]. Arterial hypertension is a common side effect of cyclosporine A (CyA). Aim of the study was to evaluate the activity of erythrocyte (RBC) Na transport in two groups of patients with a well functioning renal graft (Crs less than 1.7 mg/dl) treated by prednisone+azathioprine (10 pts), or prednisone+CyA (21 pts), in relationship with blood pressure status. Twenty-one age matched healthy subjects were studied as a control group. Na,K pump and Na,K cotransport were significantly lower in CyA than in AZA patients (2,184 +/- 106 vs 3,089 +/- 162 and 58 +/- 8 vs 187 +/- 28 mumol/l RBC/h: p less than 0.01), without differences between normotensive and hypertensive patients. Na,K pump efflux in normal subjects was 2334 +/- 66 mumol/l RBC/h (p less than 0.01 vs AZA), NA,K cotransport was 205 +/- 18 mumol/l RBC/h (p less than 0.01 vs CyA). Significant correlations were found between RBC Na,K pump activity and trough plasma CyA levels (p less than 0.02) and between systolic pressure and plasma creatinine in CyA patients (p less than 0.01). Trough plasma CyA levels were higher in hypertensive than in normotensive CyA patients (64 +/- 5 vs 46 +/- 4 ng/ml; p less than 0.01).


The W ileal reservoir: long-term assessment after proctocolectomy for ulcerative colitis and familial polyposis. This report examines the viability of the W reservoir as a reliable option for the treatment of ulcerative colitis and familial polyposis and studies W reservoir adaptation as reflected by changes in compliance and stool frequency. Since 1984, 109 patients have undergone proctocolectomy with W reservoir reconstruction. Ileal reservoir static compliance was measured in 70 and 57 patients at 2 and 12 months after ileostomy takedown and in 25 patients at 3 years. Compliance was calculated as the change in volume over change in pressure. Twenty-four-hour stool frequency decreased from 7.3 +/- 0.2 at 2 months to 4.9 +/- 0.2 at 1 year for patients with ulcerative colitis and from 6.3 +/- 0.4 to 3.4 +/- 0.4 for patients with familial polyposis (p less than or equal to 0.05). Compliance increased from 12.7 +/- 0.6 ml/mm Hg to 14.3 +/- 0.6 ml/mm Hg between 2 months and 1 year. No significant increase in compliance occurred after 1 year. Ninety-six percent of patients were continent during the day at 12 months although 10% experienced occasional minor leakage at night. Average postoperative morbidity (for example, small-bowel obstruction, anastomotic complications) was 35%. No operative deaths, pelvic sepsis, or reservoir loss occurred. We conclude that W ileal reservoirs (1) are an excellent option for ileal reservoir reconstruction, (2) have optimal functional and compliance properties versus lower capacity designs and straight ileoanal pull-through procedures, and (3) maintain stable compliance characteristics and functional reservoir volume after the initial year of adaptation.


The spermicide nonoxynol-9 does not inactivate papillomavirus. Vaginal spermicides are effective contraceptive, and are also capable of inactivating many sexually transmitted pathogens by their detergent effect on bacterial cell membranes and viral envelopes. A 5% concentration of nonoxynol-9, the most frequently used active ingredient of spermicides, was tested for its ability to reduce the transforming activity of bovine papillomavirus type 1 (BPV-1), and the infectivity of BK virus (BKV) and cytomegalovirus (CMV). Nonoxynol-9 markedly reduced the infectivity of CMV, an enveloped virus, but did not significantly affect the activity of the nonenveloped viruses BPV-1 and BKV. Papillomavirus infections are strongly implicated in the etiology of cervical cancer. The reported protective effect of vaginal spermicides against cervical cancer is very likely not mediated by direct inactivation of papillomaviruses by the spermicide.


[Epstein-Barr virus infection and immunoglobulin abnormalities in renal transplant recipients]. The relationship between immunoglobulin abnormalities and EBV infection has been investigated in 65 renal transplant patients. Immunoglobulins abnormalities were demonstrated in 44 (68%) patients, 8 of them (18%, 12% of all patients) had a monoclonal component. Up to now no lymphoproliferative disorder has been observed in these patients. All patients were EBV seropositive at the beginning of the study and in 22 of them (33%) a reactivation of EBV infection could be demonstrated. No relation has been observed between immunoglobulin abnormalities, EBV reactivation, age or sex. By contrast, a significant relation was found between EBV reactivation and immunosuppressive treatment: patients under triple therapy with Azathioprine, Cyclosporine A and Prednisone had less EBV reactivation compared to those under Cyclosporine A treatment.


Calcium transport in dentinogenesis. An experimental study in the rat incisor odontoblast. Since cellular calcium transport mechanisms during biological calcification are less known, a series of experiments were performed by in vivo as well as in vitro methodologies in the dentinogenically active rat incisor. By means of micro-electrode technique, the pH and pCa (calcium ion activity) in predentin in situ were found to be 7.0 and 2.9, respectively. It was concluded that there exists a Ca2+ion concentrating mechanism over the odontoblast layer in direction towards the mineralization front. The kinetics of this calcium flow was determined in vivo by radiotracer technique. The time for 45Ca2+ uptake into the dentin mineral phase was determined to 10-15 min. Transmembraneous Ca2+ ion pumps and channels in odontoblasts were further analyzed. The resting membrane potential of rat incisor odontoblasts was determined to -24 mV. Using ion-specific mini-electrode technique as well as fluorescence spectrophotometry, calcium channels, Ca(2+)-ATPase and Na+/Ca2+ antiports, responsible for cellular Ca2+ uptake and extrusion, were identified in the odontoblast plasma membrane. Dissected odontoblasts were subjected to subcellular fractionation. An electrophoretic uniporter and a Na2+/Ca2+ exchanger, for Ca2+ release and uptake, respectively, were demonstrated in mitochondria, whereas a Ca(2+)-ATPase was present in the microsomal fraction. Mitochondria, microsomes and whole, digitonin-permeabilized odontoblasts, were able to maintain a steady state Ca2+ activity at pCa = 6.4-6.6 in vitro. In rats treated with colchicine, the incorporation of 45Ca2+ into dentin mineral was severely altered. Similarly, administration to rats of specific calcium channel blockers strongly inhibited 45Ca2+ incorporation. Together, the results indicate that a transcellular pathway is a major route for Ca2+ ion transport during dentinogenesis, and that this may be under a relatively strict cellular control.


[Infection by human herpesvirus type 6 (HHV-6) and renal transplantation]. Human herpes virus type 6 (HHV-6) infection was serologically investigated in renal transplant recipients. Before transplantation, 75.5% of patients was seropositive for HHV-6 and no correlation with age, sex and time on dialysis was found. During the first month after transplantation 66% of patients showed a variation in serological status against HHV-6 (seroconversion or fourfold increase of antibody titer). All patients who seroconverted had received the kidney from a HHV-6 seropositive donor, furthermore, in 11/13 (84.6%) pairs of patients receiving the kidney form the same seropositive donor, both members or had HHV-6 active infection or had no infection. The frequency of HHV-6 active infection in seropositive patients is almost the same in case of seronegative or seropositive donor. Comparing HHV-6 and CMV infections, they resulted independent as CMV infection in these patients occurs in a following period (II-III month). Notwithstanding a higher frequency of kidney rejection in patients with active HHV-6 infection, no significative correlation was found.


Electrodiagnostic medicine.

Electrodiagnostic medicine. Electrodiagnostic testing examines the physiologic integrity of the peripheral nervous system. However, such testing should represent only one part of an electrodiagnostic consultation in which the entire clinical context, including the history, physical examination, laboratory studies, and electrodiagnostic testing, is considered as a whole. Although each electrodiagnostic laboratory establishes its own normal values for nerve conduction studies and needle EMG, these values should not be used in isolation. The electrodiagnostic consultation can help narrow an otherwise broad differential diagnosis, confirm a suspected diagnosis, or help define a confusing clinical picture.


Repetitive stimulation of phrenic nerves in myasthenia gravis. In the investigation of patients with myasthenia gravis, repetitive supramaximal stimulation of an affected peripheral nerve is commonly performed to detect abnormal transmission at the neuromuscular junction. A study was undertaken to determine whether abnormal transmission could similarly be detected during stimulation of the phrenic nerves. The phrenic nerves were stimulated supramaximally with surface electrodes in 13 patients with myasthenia gravis and in 16 control subjects (six control patients with diaphragmatic weakness but not with myasthenia and ten normal subjects). The amplitude of diaphragm muscle action potentials was measured with surface electrodes during phrenic nerve stimulation at frequencies of 1-5 Hz for 3-4 seconds. In five patients with myasthenia gravis, a significant decrement (15-43% decrease) occurred in the amplitude of diaphragm muscle action potential during stimulation at 3 Hz. When stimulation frequency was reduced to 1 Hz, diaphragm muscle action potentials returned to their original amplitude within 4-5 seconds. The decrement in the amplitude of the diaphragm muscle action potential was reduced temporarily in three of four patients after the administration of intravenous edrophonium chloride (Tensilon). There was no significant change (< 10% decrease) in the amplitude of diaphragm muscle action potentials during stimulation at increased frequencies either in the 16 control subjects or in eight of the patients with myasthenia gravis. A significant reduction in the amplitude of diaphragm muscle action potential occurred in five of 13 patients with myasthenia gravis during phrenic nerve stimulation at 3 Hz but in none of the control subjects. This may be a useful and non-invasive method for identifying patients with myasthenia gravis in whom weakness of the diaphragm is suspected.


Heparin potentiation of collagen-induced platelet aggregation is related to the GPIIb/GPIIIa receptor and not to the GPIb receptor, as tested by whole blood aggregometry. To determine whether heparin potentiation of platelet aggregation is related to platelet GP IIb/IIIa and GP Ib receptors, four series of experiments were performed on blood from normal volunteers. In the first experiment pretreatment with the monoclonal antibody 7E3 (MAb 7E3), which antagonizes at the GP IIb/IIIa receptor, potently inhibited the collagen-induced platelet aggregation (p less than 0.001). With heparin added to blood pretreated with MAb 7E3, the aggregation increased (p less than 0.005) to an extent similar to that when only saline was used for pretreatment. In the second experiment, monoclonal antibody 10E5 (MAb 10E5) and peptide RGDS, substances which also antagonize at the GP IIb/IIIa receptor, decreased collagen-induced platelet aggregation to an extent similar to that after pretreatment with MAb 7E3. Following pretreatment with RGDS, heparin increased platelet aggregation (p less than 0.03), while after pretreatment with antibody MAb 10E5 heparin did not enhance platelet aggregation. In the third experiment aurin, an inhibitor of von Willebrand factor and its interaction with the platelet GPIb receptor, decreased platelet aggregation dose-dependently. In the fourth experiment heparin enhanced platelet aggregation to a similar extent (p less than 0.005), regardless of pretreatment of the blood with saline, aurin or monoclonal antibody 6D1 (MAb 6D1), the latter an antagonist at the GP Ib receptor. In conclusion, the potentiation of collagen-induced platelet aggregation by heparin was not inhibited by MAb 7E3, RGDS, aurin or MAb 6D1, but was abolished by MAb 10E5, implying that the heparin effect is related to activation of the platelet GP IIb/IIIa receptor complex.


Effect of pulmonary thromboembolism on circulating neutrophils in mice. The role of free radical generation and its scavenging enzymes in circulating mice polymorphonuclear leukocytes (PMNLs) has been studied following pulmonary thromboembolism. Levels of malonaldehyde (MDA), 02- radical generation, activity of superoxide dismutase (SOD), catalase (CAT), lactate dehydrogenase (LDH), myeloperoxidase (MPO) and lysozyme were estimated in lysed neutrophil preparations. Activities of SOD and CAT were increased in neutrophils, while animals showed 60 +/- 4% thrombocytopenia. Levels of MDA in PMNLs were also elevated significantly following thrombosis. However, there was no significant change in superoxide radical generation, after thrombotic challenge, in mice neutrophils. The present study provides evidence for the involvement of free radicals in mice pulmonary thromboembolism.


Effects of two dosages of subcutaneous low molecular weight heparin (Parnaparin) and of unfractionated heparin on fibrin formation and lipolysis in acute myocardial infarction. Although low molecular weight heparins (LMWH) have been extensively investigated for the prophylaxis and treatment of venous thromboembolism in surgical environments, few data in acute myocardial infarction are available in the literature. In this study two dosages of a new LMWH, Parnaparin, and unfractionated heparin (UF) were investigated in 50 pts with acute myocardial infarction. 20 pts received UF (15.000 units, three subcutaneous injections, Group 1), 20 pts received Parnaparin (6.400 units, single injection, Group 2) and 10 pts received a higher dose of Parnaparin (12.800 units, single injection, Group 3). Similar fibrinopeptide A (FpA) levels were observed in Group 1 and Group 2. In Group 3 the dosage of Parnaparin resulted in a significant prolongation of the APTT and in lower FpA levels. Fibrin formation was decreased by Parnaparin in a concentration-dependent way, according to both the anti-Xa activity and the APTT ratio. Parnaparin did not result in a significant increase in free fatty acid concentration, in comparison with UF. Thus, Parnaparin may offer the advantage of a single subcutaneous injection in patients with acute myocardial infarction.


The thrombolytic potency of LMW-heparin compared to urokinase in a rabbit jugular vein clot lysis model. Since the beginning of the clinical use of low molecular weight (LMW) heparins their thrombolytic or profibrinolytic potency has been a matter of controversial discussions. Regarding this problem, the aim of our study was to test a LMW-heparin (Sandoparin) in an in vivo model comparing its lytic activity to unfractionated heparin and urokinase at different doses. For this purpose a newly developed short-term rabbit jugular vein clot lysis model was developed. Urokinase infused at doses of 3300, 6600 and 10,000 U/kg to control animals for one hour showed a clear dose-dependent clot lysis. Test animals were injected with a bolus of 0.5 mg/kg of LMW-heparin followed by a constant infusion of either 0.5, 1.0 or 2.0 mg/kg for one hour. A similar dose-dependent effect was observed for LMW-heparin as for urokinase. Unfractionated heparin did not exhibit a dose-dependent lytic activity in this model. No lysis was found in rabbits treated with saline. These findings suggest that the LMW-heparin tested exhibits a dose-dependent in vivo lytic activity which can be compared to clinically effective doses of urokinase, and that this activity is not present with unfractionated heparin.


Enoxaparin (Clexane, Lovenox), a low molecular weight heparin, enhances t-PA-induced coronary thrombus lysis in anesthetized dogs without inducing hypocoagulability. An occlusive coronary thrombus was obtained in barbiturate anesthetized dogs within 60 min following the angiographic placement of a copper coil into the left descending coronary artery. This thrombus persisted for the 60 min experimental period, within which the effects of i.v. t-PA (10 micrograms/kg/min for 30 min) alone or combined with i.v. heparin (0.63 mg/kg twice) or enoxaparin (1.5 mg/kg twice) were evaluated. t-PA alone achieved recanalization for 20 min in only 2 out of the 5 dogs studied. Combination of t-PA with either heparin or enoxaparin produced this effect in all the 5 dogs studied. In dogs treated with t-PA associated to either heparin or enoxaparin, the thrombus weight was smaller (decreases of 34% and 44% respectively) than in animals given t-PA alone. The plasma amidolytic activity, expressed as t-PA activity, was greater 15 min after the beginning of t-PA infusion, in dogs pretreated with either heparin or enoxaparin than in animals given t-PA alone. Conversely, during t-PA infusion, the apparent t-PA inhibitor and antiplasmin activities were no longer measurable in the plasma, but reappeared 10 min after the end of t-PA infusion. Plasma coagulation time was not modified by t-PA, but was slightly prolonged (2-fold) by enoxaparin and markedly (7-fold) by heparin on initiation of t-PA infusion. Plasma anti-IIa activity was 3-fold higher in dogs pretreated with heparin as opposed to enoxaparin. On the contrary, both compounds increased similarly plasma anti-Xa activity. In conclusion, these results indicate that enoxaparin, like heparin, enhances the thrombolytic effects of t-PA. This favourable effect occurs independently of a plasma hypocoagulable state, which was clearly produced by heparin but not enoxaparin. Its mechanism may be the significant elevation of plasma t-PA activity produced by both heparin and enoxaparin during t-PA infusion.


Hypersensitivity of diabetic human platelets to platelet activating factor. Platelet activating factor (PAF) stimulated aggregation and [32P]-phosphatidic acid (PA) production was compared in normal and diabetic human subjects in platelet rich plasma. The concentration of PAF for half maximal (50%) aggregation of normal and diabetic platelets was 50 nM and 8 nM, respectively. PAF stimulated [32P]-PA production (a metabolite of phospholipase C pathway) was also greater in the platelets from diabetic subjects. This [32P]-PA production was inhibited by the PAF receptor antagonists SRI-63441 and SRI-63675. When the levels of glycosylated hemoglobin (HbA1c) were compared with the PAF stimulated [32P]-PA production a significant relationship was observed. These studies have demonstrated for the first time that diabetic human platelets show hypersensitivity to PAF in both aggregation and [32P]-PA production compared to normal subjects. This may be a result of some modification in phospholipid turnover mechanism and is receptor mediated. Further, the relationship of the degree of aggregation and [32P]-PA production to the level of HbA1c suggest that the insulin deficiency may contribute to these effects.


Dipyridamole inhibits platelet aggregation induced by oxygen-derived free radicals. Pyrogallol (a generator of superoxide anions) caused 50% increase in platelet aggregation induced by 400 microM of arachidonic acid. Dipyridamole did not produce a statistically significant inhibition of arachidonic-acid induced platelet aggregation, but it caused 100% inhibition of pyrogallol-stimulated platelet aggregation. Ferrous salts (Fe2+) induced 34% platelet aggregation which was inhibited (79.6%) by a concentration of dipyridamole of 10 microM. Dipyridamole inhibited ferrous-induced lipid peroxidation with IC-50 values of 17.5 microM. When arachidonic acid was used as aggregating agent, the corresponding IC-50 value was 140.5 microM. These results indicate that dipyridamole prevented platelet activation induced by oxygen-derived free radicals.


LMW heparin (anti-Xa) assays for clinical monitoring and pharmacokinetic studies on the automated coagulation laboratory (ACL). Chromogenic anti-Xa activity procedures were developed for monitoring LMW heparins on the Automated Coagulation Laboratory 300 Plus (ACL, Instrumentation Laboratory) system. For daily monitoring, a "Routine" procedure was devised which allows accurate measurements between plasma levels of 0.1 and 1.0 u/ml LMW heparin. For lower levels a "Routine-Low" method was developed which assesses activities between 0.05 and 0.4 u/ml. Due to variabilities in dODs of individual baseline plasmas, levels below 0.05 u/ml might be inaccurate when pooled normal plasma is used to establish the reference curve. While levels less than 0.05 u/ml should rarely be encountered when monitoring LMW heparins for routine clinical use, pharmacokinetic studies require accurate measurements below that level. For this reason a "Research-High" and a "Research-Low" procedure was designed. For these procedures a study subject's own baseline plasma was used to establish the reference curve. The "Research-High" measures activities between 0.4 and 2.0 u/ml, the "Research-Low" between zero and 0.4 u/ml. The procedures have excellent within-run and inter-run coefficients of variation (less than 5%) and high levels of accuracies. Even inter-instrumental reproducibilities are less than 10%. Different manufacturers' LMW heparins can be analyzed by these assays. The procedures offer full automation, great cost-effectiveness due to lower reagent volumes, rapid turn-around time and great accuracy and reproducibility.


Study of low molecular weight heparin effect on the relation between anticoagulant activity and antithrombin III affinity. Low molecular weight heparin (FR-860), and conventional unfractionated heparin (UF-heparin) were fractionated by rabbit antithrombin III (AT III)-Sepharose, and the effects of each affinity fraction on the coagulation and fibrinolytic activities were investigated. FR-860 was fractionated to no-affinity, low-affinity (LA) and high-affinity (HA) fractions, and UF-heparin to LA and HA fractions. The HA fractions showed higher activities regarding the prolongation of activated partial thromboplastin time, anti-factor Xa activity and antithrombin activity compared with those of LA. The HA and LA fractions exhibited the enhancement of heparin cofactor II (HC II) activity and fibrinolytic activity in a dose-dependent manner. These results suggest that the antithrombotic activity of FR-860 is exerted through AT III and other mechanism such as HC II-mediated system.


Labeling of platelet surface glycoproteins with biotin derivatives. We describe a non-radioactive method for the labeling of platelet surface proteins, consisting of platelet protein biotinylation by means of N-hydroxysuccinimido-biotin (NHS-B) and biotin-hydrazide (H-B); NHS-B labels proteins amino residues while H-B binds to periodate-modified sialoglycoproteins. Washed platelets were biotinylated and protein bands were detected after SDS-electrophoresis and western-blot using avidin-peroxidase and luminol as substrate to enhance the signal which was then detected by X-ray film. Biotin-labeled platelet proteins were also immunoprecipitated with monoclonal antibodies against glycoproteins Ib and the IIb-IIIa complex. The use of periodate induced biotinylation is the method of choice for labeling platelet surface glycoproteins while NHS-B also labels internal proteins. The sensitivity of this new procedure is similar to that obtained with radiolabeling techniques; biotinylation does not interfere with the antigenic properties of Ib and IIb-IIIa glycoproteins.


Could non steroidal anti-inflammatory drugs be used to potentiate L.M.W.H. activity in thrombosis? (Part II). Thromboembolic diseases are one of the main cause of mortality. Heparin fractions obtained by chemical or enzymatical depolymerization of unfractionated heparin are now widely used in the prevention of those illness. However, curative dosages have bad side effects which could be avoid by the potentiation of the antithrombotic efficacy of non-active dosages. A previous study (4) has shown that a non-steroidal anti-inflammatory (NSAI) drug like Phenylbutazone could favour the antithrombotic efficacy of Fraxiparine at a very low dose. The aim of this study was then to determine if other NSAI elements could present the same or better proactive effects.


Glycerophospholipid metabolism: back to the future. It has become customary to regard the various glycerophospholipids as quite similar, and the acyl groups are considered to have little influence on the behaviour of the lipids in membranes or metabolism. Nevertheless, a number of recent observations by the authors and others indicate a high degree of metabolic compartmentation and substrate specificity with regard to the acyl substituents (acyl specificity) of glycerophospholipid metabolising enzymes in intact cells. 1. [32P]Orthophosphate and [3H]glycerol are incorporated into phosphatidylcholine (PC) and phosphatidylethanolamine (PE) of platelets and Swiss 3T3 fibroblasts with a [32P]/[3H]-ratio several fold lower than in glycerol-3-phosphate, phosphatidic acid (PA) and phosphatidylinositol (PI), suggesting distinct metabolic separation (probably by cellular compartmentation) of the glycerol and choline (or ethanolamine) branches of de novo phospholipid biosynthesis. 2. In fibroblasts the [32P]/[3H]-ratio varied 50-fold among the molecular species of PC, PE, PI and PA, which indicates that the enzymes involved in these conversions have some degree of acyl specificity. 3. In vitro assays for lipid-converting enzymes employ detergents, which affect acyl specificity of the enzymes (lipid kinases) both by their chemical nature and concentrations. 4. Thrombin stimulation of platelets causes formation of a multitude of diacylglycerol (DAG) molecular species, but only one major molecular species of PA is formed indicating that the DAG kinase may have distinct acyl specificity in the intact cell. 5. However, this specificity could also result from the net reactions of DAG kinase(s) and PA phosphohydrolase(s), which would constitute an ATP-utilising, paired regulation of the molecular species of PA and the inositol lipids on one hand, and PC, PE phosphatidylserine and triacylglycerol on the other. These findings indicate a high complexity of glycerophospholipid metabolism and a distinct acyl specificity in intact cells that are not apparent from studies in vitro. A major challenge for future research in this area is to bridge the apparent discrepancy between in vivo and in vitro observations regarding glycerophospholipid metabolism, an endeavour that will require more knowledge about the physical chemistry of naturally occurring molecular species than is available today. The most prevailing appreciation of glycerophospholipids among biological scientists to-day is that they can be distinguished functionally, topographically and metabolically only by their head groups and that they form the bilayer in biological membranes. Most of us know that the fatty acid in the sn-2 position is unsaturated and have been indoctrinated that the higher the degree of unsaturation, the greater the fluidity of the membrane.(ABSTRACT TRUNCATED AT 400 WORDS)


Silica clotting time (SCT) as a screening and confirmatory test for detection of the lupus anticoagulants. We have developed a Silica Clotting Time (SCT) test suitable to screen patients with lupus anticoagulants (LA) and compatible with photo-optical instruments. The SCT results were considered to be positive for LA whenever the clotting times were longer than the upper normal limit at low phospholipid concentration and to be confirmed when the prolonged clotting times were corrected to normal by high phospholipid concentration. We studied plasmas from healthy subjects, patients with known diagnoses of LA, patients with acquired deficiencies of blood coagulation and hemophiliacs with anti-factor VIII antibodies. The test was positive for all LA patients, and negative for all non-LA patients except 7 hemophiliacs with anti-factor VIII antibodies. Our data indicate that the SCT is a sensitive test, suitable for screening patients suspected of having LA. Its compatibility with photo-optical instruments makes it a suitable candidate to replace the kaolin clotting time. The contemporaneous performance of SCT at low and high phospholipid concentrations provides screening and confirmation in a single procedure.


[Mortality in operable lung cancer].

[Mortality in operable lung cancer]. We have retrospectively examined the medical records and prospectively studied the survival of the 50 men and 26 women who underwent surgery for primary non-small cell lung cancer at our hospital during the period 1982 to 1986. Adenocarcinoma was the predominant histologic type of tumour (55%). Pneumonectomy was performed in only 17% of the cases. Surgery was considered to be radical in 54 patients. This was not dependent on sex, histology or type of resection. 60% of the patients were alive after three years. Almost all of them had undergone radical resection. The surviving patients (at follow-up 1 July, 1990) had been younger at the time of surgery and had a lower erythrocyte sedimentation rate than those who had died. As a group, however, they had not lived longer than those who died.


[Intraoperative ultrasonography in brain surgery]. Intraoperative ultrasound was used in 33 patients. 19 lesions were primary malignant brain tumours, including 12 gliomas, three astrocytomas and four oligodendrogliomas. There were five metastases, three meningeomas, two dysembryoplastic neuroepithelial tumours and two angiomas. One patient had an arachnoid cyst, and another an infarct. The main advantage of carrying out intraoperative ultrasound examination is that it helps to localize the tumour, particularly in small subcortical lesions where the brain surface may appear normal. Ultrasound is less useful for characterizing the tumour, although the various tumours do show some differences. As a rule, the glioblastomas are inhomogeneous and poorly marginated, while low grade gliomas are more homogeneous and well marginated. Also meningeomas and metastases tend to be homogeneous and well marginated. Periofocal oedema is hyperechogenic compared with brain tissue, with an intensity between that of normal brain tissue and tumour mass. Cyst, calcification and haemorrhage are easily demonstrated.


[Chronic hepatitis C. Experience with 50 patients]. We studied 50 patients (36 males and 14 females) with chronic hepatitis C who were admitted consecutively to our medical department during the period 1987-91. Eight patients (16%) had had a blood transfusion, 17 (34%) had used intravenous drugs and 25 (50%) were "sporadic cases" with no identifiable risk factor except that at least five had been tattooed. Most of the patients had moderate symptoms, including tiredness and asthenia. Few were jaundiced. A percutaneous liver biopsy was performed in 27 patients and showed chronic persistent hepatitis in 12 of them, chronic active hepatitis in six and cirrhosis in nine. Three patients with cirrhosis died; one from hepatoma, one from an endstage cirrhosis with bleeding and coma hepaticum, and one from septicaemia.


Indication for surgery in small-cell carcinoma of the lung. While the development of chemotherapeutic agents has lead to progress in the treatment of small-cell carcinomas of the lung, the number of local recurrences still remains high. Surgery in tumors stage I and II followed by postoperative chemotherapy is the treatment of choice and has been accepted worldwide. In tumors stage IIIa, especially in T1-3 N2 we obtained good results in the projected 3-year survival using a multimodality therapeutic regime consisting of neoadjuvant chemotherapy (3 cycles preoperative) and surgery as well as postoperative chemotherapy and irradiation of the mediastinum. Projected 3-year survival was 67% in stage I tumors, 42% in stage II and with our multimodality therapeutic regime 38% in stage III a tumors.


Hormone receptors and sites of action in the kidney. Identification of hormone target sites in the nephron has been achieved in part using autoradiography, and largely with microdissection and microanalysis techniques that permit quantitative measurements of hormone binding or postbinding effects in discrete nephron segments. The nephron target sites of hormones whose intracellular second messenger is known have been located by measuring their stimulatory effect on cyclic AMP or GMP production along the nephron. These hormones include arginine vasopressin, parathyroid hormone, calcitonin, and beta-adrenergic catecholamines. In contrast, the action sites of hormones whose cellular mediators are less well understood have been identified using micro modifications of conventional binding techniques scaled down to the minute (less than or equal to 1 microgram protein) amount of tissue available. In this group are aldosterone, corticosterone, insulin, angiotensin II, alpha-adrenergic catecholamines and dopamine. Atrial natriuretic peptides and glucagon have been studied with both methods. The precise localization of hormone receptors and sites of action in the functionally heterogeneous nephron is critical for understanding the interactions between the kidney and the endocrine system in fluid volume homeostasis, blood pressure control, and in biochemical and metabolic regulation.


Phosphoinositide hydrolysis and calcium mobilization induced by vasopressin and angiotensin II in cultured vascular smooth muscle cells. The cellular action of vasoconstrictive hormones, angiotensin II (AII) and Arg8-vasopressin (AVP), on vascular smooth muscle (VSM) in cultured VSM cells from rat mesenteric artery was studied. Both AII and AVP specifically induce a transient increases in cytosolic free calcium independent of extracellular calcium or calcium channels activated by high potassium depolarization in VSM cells loaded with Fura-2. Vasoconstrictive hormones induce a dose-dependency with formation of inositolphosphates. Analysis using high pressure liquid chromatography has shown that AVP stimulates rapid and transient increases in inositol 1,3,4-trisphosphate, inositol 1,4,5-trisphosphate and inositol 1,3,4,5-tetrakisphosphate within 1 minute. Moreover, a laser-excitation fluorescence system reveals high calcium concentration sites in subsarcolemmal region. These results indicate that, unlike voltage-dependent calcium influx across the cell membrane, AII and AVP induce receptor-mediated increases in cytosolic free calcium via phosphoinositide hydrolysis creating an intracellular messenger for calcium release from intracellular calcium stores.


Case report of hepatocellular carcinoma and cytofluorometric nuclear DNA ploidy. Cytofluorometric nuclear DNA ploidy analysis was performed in two cases of primary hepatocellular carcinoma (HCC). This study was done to get the information about clinical behavior of tissues and to assess the prognosis. In both cases, almost all the liver function tests were abnormal, the tumors were thickly encapsulated, and the size of the tumors were below 5 cm in diameter. The DNA distribution pattern was categorized in two types, diploid and aneuploid with low and high ploidy, according to the mode and degree of dispersion of cells on the DNA histogram. Low ploidy pattern of histogram was found in one and diploidy pattern in another case. The serum alpha FP level was found high in case of diploidy pattern of histogram. The case of diploidy pattern of histogram died earlier than the case of aneuploid low ploidy pattern of histogram. The DNA ploidy patterns did not relate to the survival rates of patients. A preoperative cytofluorometric nuclear DNA ploidy analysis on biopsy specimens may be potentially useful to get the information about clinical behavior of tissues, to do the selective surgical procedures, and to assess the prognosis.


Incidence, histopathologic and electron microscopic features of spontaneous nephroblastomas in rats. Primary renal neoplasms in the rats are uncommon. Nephroblastoma is the only renal embryonal tumor of the rat; all other tumors are reported in older rats. The occurrence of spontaneous nephroblastoma in rats has been reported. However, metastasis from the nephroblastoma in rat is extremely rare. Data from 2669 Sprague-Dawley control rats and 1060 Fischer-344 rats were reviewed and evaluated to determine the incidence and pathology of nephroblastoma. This tumor was observed in three Sprague-Dawley rats. Metastasis was observed in the lungs and renal lymph nodes in two different rats. No case of nephroblastoma was observed in Fischer-344 rats. Detailed histopathological and electron microscopic features of these neoplasms are described and discussed.


Smokeless tobacco contains an inhibitor of prolyl hydroxylase activity. The effects of smokeless tobacco extract (STE) and various constituents of STE on prolyl hydroxylase activity were determined using enzyme extracted from chick embryos. STE inhibited prolyl hydroxylase activity in a concentration-dependent manner, but nicotine and anabasine had essentially no effect. Enzymatic activity was inhibited by zinc, but not by the other inorganic elements in STE; however, the zinc concentration in STE was not high enough to produce the observed inhibition. The inhibition by STE was diminished by increasing concentrations of 2-oxoglutarate, but not by increasing concentrations of other cofactors. Thus, STE contains an inhibitor of prolyl hydroxylase which may be competitive with 2-oxoglutarate.


Superoxide production in experimental seizures in cats. Seizures cause cerebrovascular responses similar to those seen in conditions such as acute hypertension, ischemia/reperfusion, or fluid-percussion brain injury, which are associated with the generation of superoxide. Accordingly, we studied production of superoxide in experimental seizures. Superoxide production was measured in anesthetized cats equipped with double cranial windows using the superoxide dismutase-inhibitable reduction of nitro blue tetrazolium as a measure of superoxide production. Seizures were induced by intravenous bicuculline. The contribution of hypertension associated with seizures was studied by maintaining arterial blood pressure constant by bleeding. Significant superoxide dismutase-inhibitable reduction of nitro blue tetrazolium indicative of superoxide production was found during seizures with or without control of arterial blood pressure (1.10 +/- 0.27 and 1.29 +/- 0.16 nmol/l/min, respectively). The results show that experimental seizures are associated with superoxide generation that is independent of the rise in arterial blood pressure. It is likely that superoxide generation is due to the metabolic changes that occur during seizures.


Engineered metalloregulation in enzymes. Protein engineering of metal-dependent enzyme activity is now possible due to the wealth of information available about metalloproteins. The results emerging from these studies provide insight into our understanding of the chemistry of metals in macromolecular environments as well as the biology of metal-protein interactions.


Post kala-azar dermal leishmaniasis in the Sudan: clinical features, pathology and treatment. The clinical features, pathology, immune responses, diagnosis and treatment of post kala-azar dermal leishmaniasis (PKDL) in the Sudan are described and discussed. The disease is characterized by maculopapular or nodular lesions on the face, limbs or trunk. Lesions appear during or within months after the treatment of visceral leishmaniasis, but in 2 of 19 patients there was no previous history of kala-azar. PKDL may be confused with leprosy both clinically and pathologically. Similarities and differences between the 2 diseases are discussed. Unlike visceral leishmaniasis, the peripheral lymphoid cells of patients with PKDL respond to Leishmania antigen and some are leishmanin positive. The response to intravenous sodium stibogluconate (20 mg/kg for 30 d) was reasonably good but some patients required repeated or more prolonged treatment. Ketoconazole in a dose of 10 mg/kg daily for 4 weeks had no effect on PKDL.


Vacuolar H(+)-translocating pyrophosphatases: a new category of ion translocase. The membrane surrounding the central vacuole of plant cells contains an H(+)-translocating ATPase (H(+)-ATPase) and an H(+)-translocating inorganic pyrophosphatase (H(+)-PPase). Both enzymes are abundant and ubiquitous in plants but the H(+)-PPase is unusual in its exclusive use of inorganic pyrophosphate (PPi) as an energy source. The lack of sequence identity between the vacuolar H(+)-PPase and any other characterized ion pump implies a different evolutionary origin for this translocase. The existence of the vacuolar H(+)-PPase, in conjunction with increasing recognition of PPi as a key metabolite in plant systems, necessitates reconsideration of ATP as the primary energy source for membrane transport in plant cells.


Concomitant intestinal adenovirus infection and pulmonary cytomegalovirus infection in children causing fatal enteritis and pneumonia. Three children in Bangladesh who presented with diarrhoea, cough, dyspnoea, fever, and signs of malnutrition and died in the hospital were shown at post-mortem examination to have both adenovirus infection of the intestine (by immunofluorescence) and cytomegalovirus infection of the lung (by immunoperoxidase staining). This finding of dual viral infections of the intestine and lung in patients with concomitant enteritis and pneumonia provides a basis for symptoms emanating simultaneously from these two organ systems.


The suppressive effect of deoxyspergualin on the differentiation of human B lymphocytes maturing into immunoglobulin-producing cells. Deoxyspergualin, an analog of spergualin, has been known as a novel immunosuppressive agent with strong immunosuppressive activity in in vivo experimental systems. In the present study, we examined the effect of deoxyspergualin (DSG) and methyldeoxyspergualin (MeDSG) on the proliferation and differentiation of human B lymphocytes in vitro. Highly purified B cells from human tonsil samples were isolated by Percoll density gradient from nonrosetted cells and were used as target cells. Both agents had little effect on the proliferative response of resting or activated B lymphocytes. However, they suppressed the immunoglobulin synthesis of B lymphocytes not only in a T cell-dependent, but also in a T cell-independent system. The inhibition of antibody synthesis was manifested in the early stage of B cell differentiation. Both drugs also suppressed Ig secretion, but not proliferation, of an EBV-transformed human B lymphoblastoid cell line. These results indicate that DSG and MeDSG have a selective immunosuppressive effect on the differentiation pathway of B lymphocytes.


Japanese encephalitis in Sri Lanka--the study of an epidemic: vector incrimination, porcine infection and human disease. A prospective study of mosquito vectors, porcine infection and human disease was carried out during a Japanese encephalitis (JE) epidemic in the North Central province of Sri Lanka (November-December 1987) and a subsequent non-epidemic year (1988). The epidemic involved 361 cases of human encephalitis, virologically confirmed by immunoglobulin M enzyme-linked immunosorbent assay (ELISA), and was preceded 2-3 weeks earlier by sentinel porcine seroconversion. Virus isolation and viral antigen detection (ELISA) in field-caught mosquitoes incriminated Culex tritaeniorhynchus (Giles) and Cx gelidus Theobald as the major vectors of virus transmission during the porcine amplification and human 'spill-over' phases of the epidemic. Virus was also demonstrated in Cx fuscocephala Theobald, Cx whitmorei (Giles) and Mansonia uniformis (Theobald) during the epidemic. The major difference between the epidemic (1987) and non-epidemic (1988) years was a lower vector biomass and lower rates of virus carriage in the mosquito population.


The association of kidney graft outcome with pretransplant serum IgG-anti-F(ab')2 gamma activity. Pretransplant sera of 474 kidney graft recipients were tested for IgG-anti-F(ab')2 gamma activity. The patients had significantly higher IgG-anti-F(ab')2 gamma activity than healthy controls (P = 0.0004). Serum lymphocytotoxic antibodies were correlated with IgG-anti-F(ab')2 gamma (P = 0.004), whereas CMV infection and blood transfusions were not. We found a significant association between pretransplant IgG-anti-F(ab')2 gamma activity and early and 1-year kidney graft outcome. This association was pronounced in recipients with no lymphocytotoxic antibodies. Recipients with immediately functioning grafts and a creatinine < 130 mumol/L at 1 year had strikingly higher pretransplant IgG-anti-F(ab')2 gamma activity than patients with graft failure (P < 0.0001).


Proliferation of cytomegalovirus-primed lymphocytes in bronchoalveolar lavages from lung transplant patients. Previous reports have described an association between cytomegalovirus infection and increased donor-specific alloreactivity of bronchoalveolar lavage (BAL) lymphocytes in transplanted lungs and a higher risk of bronchiolitis obliterans due to chronic rejection. We have postulated that during infection, intragraft CMV-specific lymphocytes are activated and release lymphokines that augment cellular rejection. This study deals with an analysis of CMV antigen induced proliferation of 28 BAL lymphocyte and 27 peripheral blood lymphocytes samples from 17 lung transplant patients with or without CMV infection. Kinetic studies of lymphocyte proliferation have shown that CMV infection of the lung allograft is associated with an accelerated response of BAL lymphocytes but not PBL, following in vitro exposure to CMV antigen. These findings indicate an accumulation of primed CMV-specific lymphocytes within the lung allograft during CMV infection. Evidence has also been obtained that primed CMV-specific lymphocytes may persist for months in BAL. We conclude that the CMV antigen induced proliferation assay is useful for studies of CMV infection in transplant patients.


Rejection in lung transplantation--an immunohistochemical study of transbronchial biopsies. The number, distribution, and phenotype of mononuclear cells infiltrating the allograft lung transplant were determined immunohistochemically with monoclonal antibodies directed against cellular antigens (CD3, CD4, CD8, CD22, CD25, CD16, CD56, CD68, HLA-DR) on frozen sections of transbronchial biopsies. Seventy-two transbronchial biopsies from 21 patients undergoing lung or heart-lung transplantation were evaluated histologically and immunohistologically in a prospective study. Four major results were obtained in the graft lung parenchyma: (1) whatever the histological grading of rejection, T lymphocytes expressing CD3 were present and in a significantly higher number than in control subjects (P < 0.0005); (2) there was a positive correlation between histological rejection and the number of CD3+, CD8+, CD25+, CD16+ cells (P < 0.01); (3) the CD4/CD8 ratio was inverted (0.52 +/- 0.04), with no correlation with the histological rejection; and (4) the number and location of CD3+, CD25+ cells did not correlate with CMV identification in bronchoalveolar lavage. Immunohistochemical criteria could be used for diagnosis of rejection in the management of heart-lung transplantation.


The polymerase chain reaction, a sensitive and rapid technique for detecting cytomegalovirus infection after renal transplantation. Fifty-nine renal transplant recipients were followed during the first 3 months after transplantation. Once weekly, cultures of urine and buffy coat for CMV were taken. Furthermore, peripheral blood leukocytes were examined by an immunocytochemical assay for immediate early antigens of CMV (IEA assay) and by a polymeric chain reaction for CMV DNA. Forty-four patients had a CMV infection; 23 of them were symptomatic. PCR was positive in 22 of the 23 patients with symptomatic CMV disease. For cultures of urine, buffy coat, and the IEA assay these figures were 23, 20, and 21, respectively. The PCR was the first test to become positive in 10 patients. For the cultures of urine and buffy coat and the IEA assay these figures were 0, 2, and 2, respectively. In the other 9 patients, 2 or more tests became positive at the same time. In the patient group with a CMV infection but without symptoms the PCR also had a good correlation with the other diagnostic techniques. These results together with its short processing time of 6 hr make the PCR a sensitive and rapid technique for monitoring CMV infections after renal transplantation.