Incidence of secondary acute myelogenous leukemia after treatment of childhood acute lymphoblastic leukemia. Recent reports of secondary acute myelogenous leukemia (AML) occurring in children previously treated for acute lymphoblastic leukemia (ALL) prompted a review of patients with ALL treated at the Dana Farber Cancer Institute consortium (DFCI) between 1973 and 1987. Seven hundred fifty-two of 779 children treated for ALL entered complete remission. The mean follow-up time for the 752 patients was 4.4 years. Two children had AML develop 12 and 13 months after the diagnosis of ALL, respectively. The estimated overall risk of secondary AML was calculated for the patient population as instances per 1000 patient-years of follow-up. This was compared with recent reported cases from another institution. The estimated overall risk of secondary AML was 0.61 instances per 1000 patient-years of follow-up (95% confidence interval: 0.15, 4.4). The difference between the risk of 0.61 among DFCI patients versus previously reported risk of 5.8 among a differently treated group of patients with ALL was statistically significant (P = 0.0008). No epipodophyllotoxin was used in the patients in the DFCI consortium. In contrast, an epipodophyllotoxin was used in 12 of 13 previously reported patients who had secondary AML develop. The authors concluded that the use of epipodophyllotoxins may be associated with an increased risk of having secondary AML develop in patients with ALL.


Risk factors for cancer of the tongue and the mouth. A case-control study from northern Italy. The role of tobacco and alcohol consumption and the frequency of intake of a selected number of indicator foods as causes of cancer were investigated in a case-control study conducted in northern Italy. One hundred two men with cancer of the tongue, 104 patients with cancer of the mouth, and 726 control subjects (the latter admitted to the hospital for acute nonneoplastic disease without respiratory illness) were interviewed. Similarly strong associations were observed with cigarette smoking (odds ratio [OR], 10.5 and 11.8 for current smokers versus never smokers in cancer of the tongue and mouth, respectively) and alcohol (OR, 3.4 and 3.0 for > or = 60 versus < or = 19 drinks/week). The risk conferred by pipe or cigar smoking, although based on only 12 smokers who did not smoke cigarettes, seemed, however, to be lower for cancer of the tongue (OR, 3.4) than cancer of the mouth (OR, 21.9). Selected indicator foods and beverages, including green vegetables, carrots, fresh fruits, whole-grain bread and pasta, coffee, and tea also affected the cancer risk similarly in the two sites. The beneficial influence of such foods and beverages seemed, however, to be more marked for cancer of the mouth than for cancer of the tongue. This study suggested that, although none of the differences in the effects between cancer sites was statistically significant, tobacco from pipes and cigars and the cleansing effect of some foods of plant origin and nonalcoholic beverages may influence the risk of cancer of the tongue less strongly than the risk of cancer of the mouth.


Hodgkin disease survivors at increased risk for problems in psychosocial adaptation. The Cancer and Leukemia Group B. The long-term psychosocial adaptations of 273 survivors of advanced Hodgkin disease were assessed to determine the nature and extent of problems experienced and to identify those at high risk for maladaptation. Hodgkin disease survivors were identified who initially had been treated in clinical trials within the Cancer and Leukemia Group B from 1966 to 1986, were currently disease free, and had completed treatment for a minimum of 1 year. All survivors had advanced Hodgkin disease (with disease diagnosed at a mean age of 28 years). Survivors were at a mean age of 37 years at the time of interview (6.3 years after treatment completion), and 60% were male. Survivors were interviewed over the telephone 7-10 days after questionnaires were mailed to them concerning their psychological, social, vocational, and sexual functioning. Psychological distress was found to be elevated by one standard deviation (SD) above that of healthy subjects on the Brief Symptom Inventory, and 22% met the criterion suggested for a psychiatric diagnosis. In addition, the following problems were reported by survivors to be a consequence of having had Hodgkin disease: denial of life (31%) and health (22%) insurance, sexual problems (37%), conditioned nausea in response to reminders of chemotherapy (39%), and a negative socioeconomic effect (36%). Survivors found to be at high risk for maladaptation were: men earning less than $15,000 per year or who were currently unemployed; unmarried individuals; those with serious illnesses since treatment completion; and those who were less educated. These findings suggest that including a routine assessment of these factors would help to target survivors in need of additional evaluation and treatment.


How much brushing is enough for the diagnosis of lung tumors? Bronchoscopic investigations of lung tumors require high diagnostic accuracy. Sometimes the combination of brush biopsy with cytologic and histologic examination of forceps-obtained biopsy specimens fails to diagnose tumors. Techniques with a minimum risk and low cost when repeated several times could increase the efficiency of tumor diagnosis and help to avoid rebronchoscopy. Repeated brush biopsies were done during one bronchoscopic examination in 270 patients with pulmonary neoplasias using a flexible fiberoptic bronchoscope guided by radiographic video fluoroscopy. The results of up to five brush biopsies were compared for their diagnostic sensitivity. Singly, 68-77% of the specimens showed malignant findings. With repeated brushing, the sensitivity of the diagnostic accuracy increased to 89.6%. In the periphery of the bronchial tree, the sensitivity of brush biopsy was slightly lower in bronchoscopically invisible tumors. In 222 of 242 (91.7%) patients with positive results of brush biopsy, there was agreement in the final typing of tumor morphology. For routine bronchoscopy, repeated brush biopsy should be done to obtain the highest diagnostic yield.


A phase III trial of mitomycin C alone versus mitomycin C, vinblastine, and cisplatin for metastatic squamous cell lung carcinoma. In an effort to confirm the efficacy of mitomycin C against metastatic squamous cell lung carcinoma and to compare the efficacy of single-agent therapy with a combination containing cisplatin, the authors conducted a randomized Phase III trial of mitomycin C alone versus mitomycin C, vinblastine, and cisplatin (MVP). All patients had advanced squamous cell lung carcinoma, and survival was the primary end point. There were 133 eligible patients who received either mitomycin C alone (n = 64) or MVP (n = 69). The two groups were similar with respect to performance score, disease status, age, sex, and stage. The major objective response rates were 30% (95% confidence interval [CI], 18-41%) and 43% (95% CI, 32-55%) for mitomycin C alone and MVP, respectively (P = 0.1). The median time to progression was 83 days for mitomycin C alone, compared with 119 days for MVP (P = 0.026). The median survival time was 114 days for mitomycin C and 163 days for MVP (P = 0.09). The 1-year survival rates were equivalent. Myelosuppression was the major toxicity, and there were significantly greater leukocyte nadirs with MVP therapy (P < 0.001). Mitomycin C has antitumor activity against squamous cell lung carcinoma when used alone or in combination with MVP. The regimen containing cisplatin had marginally increased activity that did not translate into a clinically significant survival advantage.


Neuropsychologic impairment in adult bone marrow transplant candidates. Long-term cognitive impairment has been reported in adult bone marrow transplant (BMT) recipients. However, the degree to which such impairment is attributable to the procedure or is a condition existing before BMT is not known. The presence, nature, and correlates of neuropsychologic impairment were investigated in 55 adult BMT candidates, all of whom had a malignant condition. Impairment was assessed using a screening battery of standardized neuropsychologic tests. Results indicated that: (1) neuropsychologic performance was associated with specific disease and treatment risk factors, in particular a history of cranial radiation or central nervous system disease treated with intrathecal chemotherapy; (2) performance on tests reflecting memory or higher cognitive processing was more likely to be impaired; and (3) the risk of impairment increased as the number of disease and treatment risk factors for cognitive impairment in the patient increased. It was concluded that neuropsychologic impairment occurs in a significant minority of adult patients before BMT. Research is necessary to determine the extent to which such impairment significantly compromises patients' abilities to: (1) make decisions regarding undergoing BMT or participating in research protocols and (2) understand and execute self-care behaviors after BMT. More broadly, greater attention should be devoted to investigating the presence of long-term neuropsychologic impairment in adult patients with cancer.


Orbital granulocytic sarcomas (myeloid sarcomas) in acute nonlymphocytic leukemia. In a series of 89 patients with acute leukemia, orbital granulocytic sarcomas were observed in 7. All these patients had acute nonlymphocytic leukemia, and they were all children. The orbital involvement usually was bilateral, and the patients had proptosis, conjunctival hemorrhage, and chemosis. Morphologic, cytochemical, and immunophenotypic analyses did not show a predilection for any particular myeloid cell type. Two patients had biphenotypic leukemias with myeloid components. The ocular manifestations responded well to chemotherapy irrespective of the hematologic response.


Spinal epidural tumor in patients with prostate cancer. Clinical and radiographic predictors of response to radiation therapy. The authors retrospectively reviewed 50 episodes of spinal epidural tumor that occurred in 42 patients with metastatic prostate cancer and were treated with external-beam radiation. Treatment response was evaluated in terms of symptoms, neurologic status, and, in most cases, reduction of tumor on repeat myelography. At the completion of therapy, 92% of treated patients experienced pain relief and 67% had significant to complete improvement on neurologic examination. Thirty days after treatment, repeat myelography was performed in 40 of the 50 cases; compared with the initial findings immediately preceding radiation therapy (RT), the results of 58% of these studies had normalized completely, results were improved in 25%, and the results had not changed in 18%. The presence of a high-grade compression fracture of the vertebral body was an indicator of poor prognosis for tumor response on repeat myelography. The ability of a patient to walk before treatment and tumor response on repeat myelography were associated significantly with improved outcome of RT and with survival. The authors conclude that RT can effectively palliate epidural lesions from metastatic prostate cancer. The prognosis for the long-term response to therapy may be indicated by pretreatment ambulatory status and posttreatment imaging of the epidural space.


Immune-mediated disease as a risk factor for canine lymphoma. Autoimmune diseases and neoplasia have been associated as occurring simultaneously in individuals. This study evaluated the association between the simultaneous occurrence of canine lymphoma and various immune-mediated diseases. The Veterinary Medical Data Program, a national disease data registry for veterinary schools, was examined. The following immune-mediated disease categories were evaluated: lupus disorders, pemphigus disorders, autoimmune polyarthritis, immune-mediated hemolytic anemia, and immune-mediated thrombocytopenia. Odds ratios with 99% confidence intervals were calculated for the occurrence of lymphoma and each of the immune-mediated disorder categories. Only dogs with immune-mediated thrombocytopenia had a statistically significantly increased odds ratio (5.61; 99% confidence interval, 4.16-7.57) for the occurrence of lymphoma versus the general population. This association still was observed for immune-mediated thrombocytopenia when stratified by age, sex, and neutering status. Dogs with immune-mediated thrombocytopenia had a greater occurrence of lymphoma than dogs without immune-mediated thrombocytopenia.


Soluble interleukin-2 receptor serum levels in mycosis fungoides. Correlation with clinical stage. In a variety of non-Hodgkin lymphomas, a correlation between soluble interleukin-2 receptor levels (sIL-2R) and clinical stage is demonstrable. In mycosis fungoides (MF), two findings raise the question as to a similar correlation: (1) a proportion of tumor cells express IL-2 receptor and (2) sIL-2R is detectable in serum. sIL-2R were measured in patients with MF (n = 88) and atopic dermatitis (AD) (n = 14) by the enzyme immunoassay technique. Patients with AD served as controls. Cases of MF were classified according to the TNM staging classification. Sera of patients with MF with stages III, IVa, and IVb showed significantly higher values than those of stage I or II and controls. Although a close and significant correlation was found between sIL-2R levels and stage of disease in MF, it is still not clear whether elevated sIL-2R levels reflect disease activity or T-cell activation due to concomitant immunologic processes.


The prognostic value of Ki-67 antigen in non-Hodgkin lymphoma of Waldeyer ring and the nasal cavity. A monoclonal antibody, Ki-67, recognizes an antigen expressed in all phases of the cell cycle, except G0, and can be used as a simple histologic marker of cell proliferation. To assess the prognostic value of the growth fraction in non-Hodgkin lymphoma of Waldeyer ring (W-NHL) and the nasal cavity (N-NHL), the authors applied Ki-67 immunostaining combined with image analysis on such lymphomas. The authors studied 29 patients (18 with W-NHL and 11 with N-NHL), applying Ki-67 to frozen sections. The number of Ki-67-positive cells in a unit area (0.044 mm2), as an indicator of proliferative activity, and the mean area per Ki-67-positive cell (microns2), as an indicator of DNA content, were measured by the image processing system. High-grade lymphomas showed a significantly larger number of Ki-67-positive cells than intermediate-grade lymphomas (102.5 +/- 21.6 in high-grade and 46.8 +/- 8.92 in intermediate-grade lymphomas, P = 0.03), even when analyzed separately by immunophenotypes. A large mean area per Ki-67-positive cell was associated significantly with a T-cell phenotype (36.3 +/- 7.69 microns2 in T-cell lymphomas and 19.4 +/- 2.33 microns2 in B-cell lymphomas, P = 0.034) and an unfavorable clinical outcome. High proliferative activity, defined as nuclear Ki-67 expression in 2000 or more B-cell lymphoma cells and 1000 or more T-cell lymphoma cells in a 1-mm2 area, was found to be a strong predictor of poor survival among these patients (P = 0.048 and P = 0.009, respectively). Ki-67 immunostaining, combined with image analysis, is a novel method for determining a tumor proliferative index that provides useful clinical data regarding head and neck lymphomas.


The future of prognostic factors in outcome prediction for patients with cancer. The anatomic description of the extent of tumor spread (tumor staging) assists clinical management, facilitates communication among physicians, is an essential part of randomized controlled trials, and may help in the counseling patients and their families. However, in recent years, additional "prognostic factors" have been defined, many of which assess or reflect the biologic behavior of malignant neoplasms. Other measures of tumor biochemistry address the natural history of neoplastic development and often are included in a discussion of new prognostic factors. This review article summarizes current knowledge and thinking related to tumor prognostic factors in four areas by providing: (1) a definition and principles of anatomic spread of tumor (staging) and some suggestions for improvement, (2) a description of some examples of additional factors of prognostic significance, (3) some statistical methods to evaluate prognostic factors, and (4) an examination of the possible future of summary statements of outcome (i.e., prognostic indexes).


[Patients and solicitude].

[Patients and solicitude]. Patients are not only passive and vulnerable beings who need to be cared for. They are also autonomous, active and capable of caring for others as well as for themselves. In this study, the author has completed lengthy interviews with five women in order to identify different types of caring. Complicity-based caring is characterized by verbal and nonverbal behavior of patients who are sharing a common experience. This type of caring can be encouraged by a third-party who brings patients together with similar experiences. Action-oriented caring consists of intervening for other patients as an antidote for one's own fear, panic and anguish. Solidarity-based caring corresponds with assisting fellow patients in finding and maintaining their own identity, pride and dignity. Co-operative caring can be observed in group-type settings such as the behavior observed during support groups or vegetarian cooking classes for cancer patients. Self-centred caring refers to the various forms of concern one has for one's self. The experience of suffering can determine how caring is expressed. Concern can force individuals to examine their way of acting and interacting more carefully. Conscious choices are made to avoid empty and exhausting relationships and encourage the growth of positive relationships. Paradoxically, this change in perspective brings about an open mind and heart. It is at this time that caring for healthy individuals appears. Unfortunately, there is very little distinction between patients and healthy individuals, but rather sensitive relationships between individuals facing difficult life challenges.


[Cohabitation of lucid and non-lucid residents]. The primary goal of this study was to confirm the appropriateness of an experimental model that studied the emotions of a rational client toward living with cognitively impaired clients. The second goal was to identify factors linking the feelings of health caregivers toward the cohabitation or segregation of rational and cognitively impaired couples. The research took place in 19 Montreal senior citizen homes with 75 beds or more. The senior citizen homes, the rational clients and the health caregivers were selected using specific criteria and systematic sample selection. Individual interviews with 435 rational clients living in cohabitation units were undertaken. In addition, 349 health caregivers (nurses, practical nurses and attendants) responded to a questionnaire relating to the cohabitation of these couples. It was found that the rational clients adjusted well to the model. Four variables reflecting the emotions of 33 per cent of the rational partners include: The frequency of uncomfortable feelings related to living with the cognitively impaired; their knowledge level about cognitive impairment; their distress level relating to the confused/irrational behavior of the cognitively impaired; and their perception of the advantages of living with the cognitively impaired. The rational clients who had the least understanding of cognitive impairment identified that they were more upset by the confused client's behavior. They demonstrated uncomfortable feelings toward living with confused/irrational clients more frequently, and perceived less advantages relating to cohabitating with these clients. Consequently, the rational clients were less amenable to cohabitation. The percentage of cognitively impaired clients living on the same floor was not identified as a variable.(ABSTRACT TRUNCATED AT 250 WORDS)


A medical workstation for the evaluation of alternative 3D radiotherapy treatment plans. A medical workstation for the evaluation of alternative 3D radiotherapy plans is described. Our system can simultaneously apply qualitative as well as quantitative evaluation methods to as many as three 3D dose distributions. The user interface of the workstation has been designed with the demands of radiation oncologists in mind. The handling of the large amounts of 3D data (CT data, up to three 3D dose distributions, volumes of interest) is assisted by computer graphics.


Variability of the regional cerebral blood flow pattern studied with [11C]-fluoromethane and position emission tomography (PET). The mean regional cerebral blood flow (rCBF) pattern measured with [11C]-fluoromethane and positron emission tomography (PET) in 26 healthy subjects was heterogenous throughout the brain showing the highest rCBF in the medial prefrontal cortex and the lowest rCBF in the inferior temporal cortex. Right/left asymmetry of the mean rCBF was not significant. The variability of the rCBF pattern was assessed by dividing the subjects into one group of naive subjects and one group of subjects who had habituated to the scanning procedure. Naive subjects had a significantly higher mean rCBF (p less than 0.05) in defined areas of the higher order association cortices predominantly in the right cerebral hemisphere, but a virtually identical mean rCBF in the primary cortical input and output areas. These findings suggest a raised level of mental activity in subjects undergoing the first PET measurement.


Study on the change of lung lamellar body to lattice tubular myelin by N-acetylglucosaminidase with special reference to membrane components and calcium. The possibility of a role of N-acetylglucosaminidase or beta-galactosidase in the morphological change of lung lamellar body to lattice tubular myelin was examined in an in vitro incubation study. Electron microscopic observation of the incubation product and gas-chromatographic analysis of released sugars during incubation were used to understand the mechanism of these enzymes in the change of membrane constitution. N-acetylglucosaminidase proved to play a role in developing membrane of lamellar body toward lattice tubular structure with the support of Ca++. In this process, inositol release was indispensable in the modulation of membrane as well as the release of a small amount of N-acetylglucosamine. However, further reaction of this enzyme caused destruction or fusion of lattice-tubular integration with the release of other membrane components. Beta-galactosidase played no role in this process.


Myofibrosarcoma of subcutaneous soft tissue of the cheek. A low grade soft tissue sarcoma from the naso-labial fold and designated as myofibrosarcoma (sarcoma of myofibroblasts) is described using standard light microscopy techniques, immunohistochemistry and electron microscopy. Pink fibrillated cytoplasm, regarded as one of the typical histological features of leiomyosarcoma, was not obvious but immunostaining for alpha-smooth muscle actin was positive suggesting smooth muscle differentiation. By electron microscopy, tumour cells contained abundant rough endoplasmic reticulum cisternae and a large Golgi body; there were modest but numerous bundles of fine filaments with focal densities. A conventional lamina was lacking but the cell surface was characterised by fibronexus junctions in which there was a conspicuous extracellular, fibronectin-containing fibril, apparently mediating contact between cell and matrix. The tumour cells therefore showed myofibroblastic differentiation. The cell surfaces showed strong immunoreactivity with an anti-fibronectin antibody. Myofibrosarcoma is not a widely recognised entity and this case is only the 7th example to be documented in detail by means of both immunohistochemistry and ultrastructure. The combination of electron microscopy for detecting the fibronexus junction and immunostaining for fibronectin at the cell periphery is suggested as potentially useful for distinguishing myofibrosarcoma from leiomyosarcoma.


Applications of hierarchical image segmentation techniques: aorta segmentation. The segmentation of objects from complex images is difficult due to indistinct boundaries between objects and similarity of objects. We have used a hierarchical segmentation approach to accurately distinguish between objects and identify the corresponding boundaries. This approach has been used successfully to extract the aorta from transverse magnetic resonance (MR) images of the abdomen. The procedure to segment the abdominal aorta involves three progressive steps: aorta detection, aorta extraction, and estimation of the aorta wall boundary. Comparison of hierarchical segmentation techniques with single-step segmentation methods (e.g., region-growing, edge-detection) shows that hierarchical segmentation yields more reliable results.


Quantitatively evaluated ultrastructural findings can add to the differential diagnosis between keratoacanthoma and well differentiated squamous cell carcinoma. The differential diagnosis between keratoacanthoma (KA) and well differentiated squamous cell carcinoma (WDSCC) is not always easy to perform. Seven cases of KA and seven cases of WDSCC have been here analyzed by morphometry on ultrastructural sections and compared with normal epidermis. Parameters expressing the cohesivity among epithelial cells (numerical and surface density of desmosomes; volume density of intercellular space) were significantly different in KA and WDSCC, so that they may be useful in differential diagnosis. The Authors also questioned the nature of KA, suggesting a continuum between this lesion and WDSCC.


Isolation of plasma membranes, Golgi bodies and mitochondria of Xenopus laevis morulae. Identification of plasma membrane proteins. Homogenates of Xenopus morulae at the 16-32 cell stage were centrifuged on discontinuous sucrose gradients. Isolated fractions were identified by electron microscopy (EM) as mitochondria, a fraction enriched in Golgi vesicles, and plasma membranes. A special effort was made to prepare plasma membranes free of cytoplasmic contaminants. The resulting purified plasma membranes appeared morphologically identical to plasma membranes in situ. The external surface is covered with a fibrillar coat while vesicles are seen attached to their inner surface. O'Farell's method (1975) was used to obtain protein patterns of the various fractions on 2-dimensional gel electrophoresis. Each fraction displayed a specific pattern. By comparing the different patterns, it was possible to identify a group of proteins as belonging to the plasma membrane fractions. Labelling of cell surface with sulfo-N-hydroxysuccinimido-biotin together with differential extraction of proteins has allowed us to tentatively allocate these proteins in different structures of the plasma membrane fractions. The data presented in this paper corroborate and extend our ultrastructural studies on neogenesis of interblastomeric plasma membranes (Bieliavsky and Geuskens, 1990).


Metastatic pineoblastoma via a ventriculoperitoneal shunt: CT demonstration. We report the CT findings in a patient with pineoblastoma metastatic to the peritoneum via a VP shunt. A large, soft tissue tumor mass was revealed in the pelvis with associated peritoneal seeding and ascites. The initial intracranial tumor biopsy and later biopsy of metastatic peritoneal tumor demonstrated identical tissue diagnostic for pineoblastoma. Patients with intracranial malignant neoplasms and VP shunts will be followed during or after treatment with MRI brain scans. Periodic CT abdomen scans should be obtained to detect potential peritoneal metastases early to allow more effective treatment.


Image segmentation in digital mammography: comparison of local thresholding and region growing algorithms. Local thresholding and region-growing algorithms are developed and applied to digitized mammograms to quantify the parenchymal densities. The algorithms are first evaluated and optimized on phantom images reflecting varying image contrast, X-ray exposure conditions, and time-related changes. The difference between the segmentation results of the two techniques is less than 6% on the phantom images and 11% on the mammograms. The agreement between the computerized procedures and a manual one is in the range of 74-98%, depending on the breast parenchymal pattern and segmentation algorithm. The results show that computerized parenchymal classification of digitized mammograms is possible and independent of exposure.


Age-related quantitative changes in inhibitory axo-somatic synapses on Purkinje cells of rat neocerebellum (Crus I and Crus II). Morphometric analysis concerning inhibitory axo-somatic synapses on the somata of rat Purkinje cells was carried out. The percentage and the absolute cell surface area occupied with the synaptic junctions as well as their surface-to-somatic volume ratio exhibited significant changes with ageing (from 2 to 24 months). All the parameters revealed the smallest value at 2 months and a peak at 9 months; then, after a decline towards 12 months, they remained fairly constant until 24 months. It is suggested that the observed modifications in the synaptic parameters of the somata would have some connection with compensatory mechanisms for dendritic deafferentation seen with ageing or with the onset and development of the reproductive life of the animals.


Massive putaminal-thalamic nontraumatic hemorrhage. Fourteen patients developed massive putaminal-thalamic hemorrhage. All patients were young black men. They were hypertensive but without chronic hypertensive vascular changes. They had been treated with antihypertensive medication for less than 3 yr. All patients presented with a prodromal headache beginning 18-30 h before the brain hemorrhage. Initial clinical signs were heralded by a change in the headache pattern and vomiting. All patients became comatose and hemiplegic within 4-12 h. CT showed a hyperdense putaminal-thalamic hemorrhage which was 60 to 86 mm in maximal diameter. There was marked mass effect with secondary intraventricular extension. All patients died within 72 h, despite rapid and adequate blood pressure control and maximal medical treatment of cerebral edema and increased intracranial pressure.


Case report: extramedullary plasmacytoma of the larynx. A case of extramedullary plasmacytoma of the larynx is presented. The case is unique for the extramedullary plasmacytoma's marked extension into the mediastinum. The computed tomography (CT) features are described.


Synchronized shift in localization of the Golgi complex and the microtubule organizing center in the terminal phase of cytokinesis. As mammalian cells enter mitosis, the Golgi complex is disorganized and the remnants are dispersed throughout the cytoplasm in the form of a few short cisternae and small clusters of vesicles. Once the separation of the chromosomes is completed and nuclei reform, stacks of flattened cisternae reappear and a united Golgi complex of interphase type starts to be reorganized. This process is believed to ensure an approximately equal partitioning of the Golgi complex on the daughter cells. Here, the configuration of the Golgi complex and its relation to the cytoplasmic microtubule system was studied at the end of cytokinesis using synchronized cultures of L929 mouse fibroblasts and rat dermal fibroblasts. One hour after the release of the mitotic block, the Golgi complex (visualized immunocytochemically with antibodies against mannosidase II) was most frequently located on the proximal side of the nucleus as related to the intercellular bridge (visualized immunocytochemically with antibodies against tyrosinated alpha-tubulin). One hour later, it was preferentially found on the distal side of the nucleus as related to the intercellular bridge. Immunocytochemical demonstration of the radiating pattern of microtubules, and direct demonstration of the centrioles using antibodies against detyrosinated or acetylated alpha-tubulin, showed that the microtubule organizing center (MTOC) shifted position in a similar manner as the Golgi complex. Moreover, double staining with antibodies against mannosidase II and tyrosinated alpha-tubulin revealed that the Golgi complex and the MTOC codistributed at both times after the release of the mitotic block. Electron microscopic analysis confirmed that the reforming Golgi stacks first gathered close to the centrosome (a pair of centrioles with associated structures, constituting the main MTOC in the cell) on the proximal side of the nucleus and that the Golgi stacks and the centrosome were subsequently both relocated to the distal side of the nucleus as related to the intercellular bridge. Taken together, the findings indicate that the Golgi complex goes through a characteristic translocation in the terminal phase of cytokinesis and confirm the idea that the cytoplasmic microtubule system plays an important role in the organization of this organelle system. A possible function of the shift in location of the Golgi complex at the end of cytokinesis could be to direct membrane traffic first to the elongating intercellular bridge and thereafter to the leading edge as the cells are about to separate and move away from each other.


MRI diagnosis of biceps tendon rupture.

MRI diagnosis of biceps tendon rupture. Distal biceps tendon rupture is a rare injury. While plain radiographs are usually not helpful in visualizing the defect, evaluation of the tendon with MRI has proven to be useful as it depicts complete anatomic detail of the tendon and associated structures. Its multiplanar imaging capability and superior soft tissue contrast resolution makes MRI a prime diagnostic tool in the diagnosis of tendon disorders. We report a case of complete rupture of the biceps tendon diagnosed by MRI.


Subcellular structure of the atrial myocardium of children in cases of atrial septal defect. In order to study the initial development of myocardial ultrastructural changes owing to right atrium volume overload, myocytes have been studied in specimens taken from the right atrial wall and auricle of four children aged 1 to 6 years with ostium secundum atrial septal defect undergoing cardiac surgery. The younger patients (1 to 4-year-old children) we observed did not show diffuse and significant myocardial ultrastructural damages. The most significant myocardial changes were observed in the 2 older patients (six years old) as we found subcellular signs of myocardial hypertrophy such as an increased number of mitochondria, increased glycogen inclusions, areas of new sarcomerogenesis and nuclei lobulated and variably shaped. Focal degenerative changes, such as rupture of mitochondrial cristae and intercellular fibrosis were also noted. These changes may be considered as the initial features of myocardial hypertrophy because they were not as severe and diffuse as those usually seen in a marked functional failure.


Rickettsiae-like microorganisms in the midgut and other visceral tissues during development of Drosophila auraria. Rickettsiae-like organisms (RLOs) were identified for the first time in midgut, Malpighian tubules, wreath cells (ventral nephrocytes), spermatogonia and gut muscles in a species of Drosophila. Their number in the midgut cells of Drosophila auraria significantly increased at the late third larval instar and at the beginning of pupation. The RLO population in the larval midgut followed the fate of their host cells and was destroyed during metamorphosis. The RLOs pass from the larval to the adult midgut via the RLOs existing in the 'regenerative cells', which will form the adult midgut during population. The total volume of RLOs per anterior midgut cell increased analogically in relation to the absolute volume of the host cells till the 130 h larval stage. However, during the late-third instar and at the beginning of pupation the total volume of RLOs per host cell significantly increased. The physiological significance of the latter data as well as the relationship between RLOs and host cells are discussed.


Early ultrastructural changes during thioacetamide-induced apoptosis in rat liver. An histological and ultrastructural study of the early changes in the liver following a single administration of thioacetamide (TH), was carried out in male Wistar rats. One hour after treatment, apoptosis was already present in the liver. By electron microscopy, the following sequential changes were observed: progressive detachment of hepatocytes from neighboring cells, formation of surface infolds with multiple blebs and, finally, release of several membrane-bounded apoptotic bodies in the extracellular space and into the sinusoidal lumen. Three hours after TH administration, the apoptotic cycle was almost entirely completed, as shown by the presence of phagocytosed apoptotic bodies inside the cytoplasm of intact liver cells. Our study evidences that TH induces apoptosis of liver cell as early as one hour after its administration. Moreover, our data show that the apoptotic cycle may be completed in 3-4 h. From the morphological point of view, apoptosis induced by TH appears indistinguishable from programmed cell death, occurring during embryogenesis or metamorphosis, and from apoptotic cell death seen during regression of mitogen-induced rat liver hyperplasia.


Cytochemical characterization of the cuticle of Caenorhabditis elegans (Nematoda: Rhabditoidea). At the ultrastructural level, the Caenorhabditis elegans (Maupas, 1900; Doughert, 1953) cuticle shows the presence of six layers: epicuticle, external cortical, internal cortical, intermediate, fibrous and basal. Two techniques were used for carbohydrate localization: the periodic acid-thiosemicarbazide-silver proteinate (Thiéry) and gold-labelled lectins. No labelling was found on the nematode's cuticle. With the ethanolic phosphotungstic acid technique (E-PTA), that detects basic proteins, reaction product was observed in the outer cortical layer, in the cuticle struts and in the dense bodies of the muscle cell. Surface anionic sites of C. elegans were visualized by using cationized ferritin particles, at pH 7.2, and by using colloidal iron hydroxide particles at pH 1.8. Treatments with trypsin and neuraminidase (Vibrio cholerae) did not interfere with the binding of the cationic particles to the nematode's surface. In contrast, treatment with chondroitinase ABC, a specific enzyme for glycosaminoglycans, significantly reduced the binding.


Characterization of four melanoma cell lines with electron microscopy, immunocytochemistry, cytogenetics, flow cytometry, and southern analysis. Four cell lines established from human metastatic malignant melanoma, derived from four patients, were analyzed. Ultrastructurally and immunocytochemically, the cultured tumor cells had retained characteristic features of melanocytes and of the primary malignant melanomas. The genetic stability was investigated by repeated flow-cytometric and cytogenetic analyses over 24 months of continuous cultivation. The DNA indices ranged from 1.7 to 2.1 and were stable during the entire period. The same was true for the karyotypes, which had modal numbers ranging from 50 to 84. The most common types of abnormalities were: isochromosomes i(1q), i(9q), translocations (1;17) and (3;6), and other aberrations (1p+,4p+,5p+,11p+,11q-,11q+). Abnormalities involving chromosome 1 were present in all cell lines, but loss of genetic material from chromosome 1p was demonstrated in only one of four cell lines when tested by the Southern blotting technique using a lambda MS1 probe.


Karyotype evolution in a patient with biphenotypic neonatal leukemia. We present the case of a 4-day-old boy with acute lymphoblastic leukemia showing at onset a karyotype 46,XY,t(4;11)(q21;q23). At relapse an additional change, add(2), was present. Molecular analysis showed the same immunoglobulin rearrangement both at onset and at relapse, but immunohistochemical analysis revealed some cells having myeloid features. A continuous cell line derived from the leukemic blasts of the patient presented typical monoblastic features.


Cytogenetic and FISH studies of abnormal X chromosomes in a patient with ANLL. We report a case of idic(X)(q13),r(X)(p22q13), and del(X)(:p11-->cen-->q11:) in a 71-year-old female patient with de novo acute nonlymphocytic leukemia (ANLL), FAB-M4. The abnormal X chromosomes of this patient were identified cytogenetically by G-banding technique and were further confirmed by fluorescence in situ hybridization (FISH) using an alpha-satellite probe to chromosome X centromere. The features of this are compared with other cases reported in the literature.


Temporal association of marrow eosinophilia with inversion of chromosome 16 in recurrent blast crises of chronic myelogenous leukemia. We report a patient with Ph+ chronic myelogenous leukemia (CML) whose recurrent blast crises were associated with marrow eosinophilia and inv(16). After intensive chemotherapy, for each blast crisis, the patient reentered chronic phase with disappearance of both the inv(16) and the eosinophilia.


Detection of an i(17q) chromosome by fluorescent in situ hybridization with a chromosome 17 alpha satellite DNA probe. An isochromosome for the long arm of chromosome 17,i(17q), is frequently found as an additional chromosome aberration to the Ph with advanced disease in the chronic myelocytic leukemia (CML). We studied an i(17q) in blood samples from two patients with CML in blast crisis with a biotinylated chromosome 17 specific alpha satellite deoxyribonucleic acid probe. G-banded karyotypes of these patients showed a dicentric i(17q), dic(17)(p11.2). Fluorescence in situ hybridization (FISH) delineated one normal chromosome 17 and one i(17q) among metaphase chromosomes; the latter showed a dicentric pattern. In most interphase nuclei of both patients, two fluorescence spots were observed. In some interphase nuclei, including mature neutrophils, the dicentric chromosome was discernible by its size and shape of the fluorescent spots. Three fluorescent spots were observed in a small proportion of interphase cells, and existence of a subclone with two normal chromosome 17 and an i(17q) was confirmed by examining a large number of metaphase plates. The results of FISH provided us with information of numerical and structural aberrations of chromosome 17 in interphase cells.


Significance of trisomy 7 in thyroid tumors. Standard cytogenetic studies of a multifocal metastasizing papillary thyroid carcinoma revealed two clonal chromosome aberrations: rearranged 10q and trisomy 7. Trisomy 7 seemed to be restricted to tumor nodule A, whereas era (10q) was detected in tumor nodule B and in a metastatic lymph node. We applied fluorescent in situ hybridization to ask whether trisomy 7 was a feature of the original tumor nodule or an in vitro phenomenon changing quantitatively during early passages and to see whether trisomy 7 was restricted to tumor nodule A. We used the biotinylated chromosome 7 alpha-satellite probe D7Z1 on freshly dropped slides from metaphase harvests from tumor nodule A,B, and the lymph node and on touch preparations from the frozen specimen of tumor nodule A. Trisomy 7 was present in the original tumor nodule (6% of cells), as well as in early passages (P1-3) from both tumor nodules and the metastatic lymph node with a frequency of 10.7-13.2%. The detection of trisomy 7 as a stable component in short-term cell culture and its presence in the original tumor material indicates that this common numerical aberration is an in vivo phenomenon.


Microprocedure for in situ nick translation of chromosomes. We have modified the procedure of in situ nick translation to shorten the autoradiographic exposure time from 1 month to 3 days and reduce the volume of nick translation solution by a factor of at least 10. The modified procedure can be carried out on individually chosen chromosome spreads. The procedure was used on chromosome spreads of three related lines of mouse mammary epithelium (+SA, -SA, CL-S1) with different degrees of tumorigenicity. We found that the autoradiographic silver grains that are observed following in situ nick translation were often placed at the apparent junction site of chromosome translocations or at the breakpoint of chromosomal pieces. We found also that silver grains were located above double minute chromosomes, which suggests that there are active genes in double minutes.


Cytogenetic analysis of hematologic malignancies in Hong Kong. A study of 98 cases. The karyotypes of 98 patients between the ages of 8 and 81 years with acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), and chronic myeloid leukemia (CML) are presented. Although the well-described cytogenetic abnormalities associated with particular FAB subtypes in the West were observed, certain important local differences were noted. In ALL, hyperdiploidy was rarely observed, whereas the Philadelphia chromosome was observed in 50% of abnormal karyotypes. In AML, the t(8;21) was infrequently observed in M2 case, whereas trisomy 4 and 6, rarely reported elsewhere, formed 12% of the abnormal cases. In MDS, the incidence of -5/5q- and/or -7/7q- was 83% of cases with aberrant cytogenetic findings. Neither i(17q) nor an extra Ph was seen in 26 cases of CML including 9 cases of accelerated phase/blast crisis. In addition, previously unreported cytogenetic abnormalities occurring as single cases are presented. These findings are discussed in the context of geographical heterogeneity of chromosomal abnormalities in leukemia and emphasize the importance of continued epidemiologic studies of cytogenetics in hematologic malignancies.


Longitudinal cytogenetic study of metaphase and interphase cells in childhood monosomy 7 syndrome. A 15-year-old male with myelodysplastic syndrome (MDS) characterized by monosomy 7 was cytogenetically evaluated by metaphase karyotyping and fluorescence in situ hybridization (FISH) of interphase cells at six different points during the course of his disease. At diagnosis, there was complete agreement between metaphase and interphase findings. Interphase analysis alone provided important cytogenetic information on the first specimens received following intensive combination chemotherapy and bone marrow transplantation where metaphase analyses were uninformative. The detection of a minor post-treatment monosomy 7 population by interphase but not metaphase studies may have identified minimal residual disease prior to recurrence of MDS. From this longitudinal study, it is concluded that metaphase and interphase cytogenetic analyses form complementary approaches and that use of both provides greater analytical power when appropriate chromosome markers are available.


Isolation of a yeast artificial chromosome clone that spans the (12;16) translocation breakpoint characteristic of myxoid liposarcoma. Cytogenetic analysis of liposarcomas has demonstrated that translocation (12;16) (q13.3;p11.2) is characteristic of the myxoid subtype of this adipose tissue tumor. Our previous results suggested that the GLI gene is close to the translocation breakpoint on chromosome 12. We now describe a yeast artificial chromosome (YAC) that contains GLI and spans the chromosome 12 region involved in the t(12;16) breakpoint. This clone will permit rapid definition of the genetic region surrounding the breakpoint and allow isolation of the gene presumably affected by the translocation.


Interphase cytogenetics on paraffin sections of malignant pleural mesothelioma. A comparison to conventional karyotyping and flow cytometric studies. We performed in situ hybridization (ISH) studies of malignant pleural mesotheliomas to detect numerical aberrations of chromosomes 1 and 7 in interphase nuclei of paraffin sections of 13 cases that had been analyzed previously by conventional karyotyping and flow cytometry. The hybridizations were performed with the biotin-labeled probes recognizing repetitive DNA sequences in the (peri)centromeric regions of chromosomes 1 (1q12) and 7(7cen). Application of histologic sections allowed us to analyze the tumor cells only. Comparison of the karyotype and ISH studies showed that the same chromosome copy numbers were detectable by both methods in 13 (chromosome 1) and in 12 (chromosome 7) cases evaluable by ISH. DNA indexes determined in the paraffin-embedded tumor material corresponded with the ISH findings. As compared with karyotype analysis, ISH showed a larger heterogeneity in chromosome copy numbers. The results can be divided into three groups: 1) Monosomy or disomy of chromosomes 1 and 7 was detected by both methods in two cases; 2) in four cases, disomy of both chromosome 1 and 7 was observed in most of the cells by ISH analysis, and karyotype analysis had shown clear polyploidization in three of these cases; 3) in seven cases, supernumerary copies of chromosomes 1 and/or 7 were present in an evident fraction (27-80%) of the cells analyzed by ISH, and karyotype analysis confirmed the aberrant copy numbers in five of these cases. On the other hand, ISH showed copy numbers not detected by karyotype analysis in six of the seven cases. Thus, by combining karyotype and interphase cytogenetic studies, complementary information about chromosomal aberrations in mesothelioma is obtained.


Analysis of prostatic tumor cultures using fluorescence in-situ hybridization (FISH). Analysis of ten primary prostatic tumor cultures using fluorescence in-situ hybridization (FISH) with pericentromeric probes for chromosomes 7, 8, 10, 16, 17, and 18 revealed aneusomies in nine of these specimens. Classical cytogenetics by G-banding indicated that only four of those same ten specimens had any (but not consistent) clonal abnormalities. This preliminary study suggests that aneusomy is a common event in early-stage prostatic tumors, and also supports the notion that multiple chromosomes are involved. In combination with routine cytogenetic analysis, FISH is thus likely to be a powerful tool in the evaluation of prostatic cancer.


Complex chromosomal rearrangements in an unusual variant of hairy cell leukemia. An unusual case of variant HCL with multiple ribosomal lamellar complexes and complex chromosomal changes is described. The karyotype of the main cell clone is characterized by involvement of chromosome 5 at q13.3 and interstitial deletion of 7q. However, there is no other evidence of myelodysplasia and the abnormal cells are of lymphoid origin with a B-cell immunophenotype. The clonal chromosomal evolution involves rearrangements at sites known to be specifically altered in lymphoid tumors.


t(13q;17p) and del(5q): possibly specific changes in Chinese patients with colorectal cancers. Cytogenetic study of 18 colorectal carcinomas confirmed the extensive heterogeneity and the complexity of the karyotypic picture in this type of tumor. Karyotypic analysis showed that chromosomes 17p and 5q, in both numerical and structural aspects, were the most frequently involved chromosomes and prone to losses. The most common structurally rearranged forms were translocations of 17p with other chromosomes, especially t(13q;17p), which constituted over 50% of all 17p rearrangements, and an interstitial deletion of 5q that made up as much as 73% of all structural abnormalities of 5q. According to the results, we conclude that chromosomes 17 and 5 may play important roles in the evolution of colorectal cancer and t(13q;17p) and del(5q) may be possibly specific to Chinese patients with colorectal cancer.


Acute lymphoblastic leukemia of Burkitt type (L3) with a (14;18) and an atypical (8;22) translocation. A 67-year-old man had night sweats, tumor lysis syndrome, and typical histologic and immunophenotypic features of Burkitt-type acute lymphoblastic leukemia (ALL). Cytogenetic analysis showed a previously unreported karyotype, a combination of a (14;18) and an atypical (8;22) translocation.


Cytogenetic analysis of a uterine lipoleiomyoma. We cytogenetically analyzed a uterine lipoleiomyoma. A primary chromosomal abnormality, t(12;14), was found in all 62 cells studied. A secondary change involving chromosomes 1 and 5 was detected in 15 of 62 cells. These findings suggest that lipoleiomyomas share the same chromosomal abnormalities found in common leiomyomas. We speculate that the secondary chromosomal change involving chromosomal 5 may be responsible for the lipomatous change.


Translocation (5;22) in an Askin's tumor. A boy with a brain metastasis of an Askin's tumor was investigated. Cytogenetic studies revealed a near-diploid karyotype with monosomies of chromosomes 5 and 22 in the presence of a derivative chromosome der(5)t(5;22)(q35;q11). In particular, no involvement of chromosome 11 was seen.


Antimutagenic effects of polyphenolic compounds. Smokers expose themselves to potent carcinogens daily. One of them is the nicotine-derived nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). Since estimates are that humans consume 1 g of phenolic compounds/day, we investigated the inhibitory effects of five structurally related polyphenolic compounds on the mutagenicity of NNK in Salmonella typhimurium TA1535. NNK at a concentration of 80 mM was activated by hamster liver microsomes. The antimutagenic efficacies were dose-related between the non-toxic concentrations of 0.1 and 0.5 mmol/dish in the following order: esculetin > ellagic acid > (+)-catechin > propyl gallate > (-)esculin. At the highest non-toxic dose tested (0.5 mmol/dish), these polyphenolics inhibited mutagenesis in TA1535 by 77%, 67%, 62%, 59% and 53%, respectively. The results of this study demonstrated that polyphenolic compounds may inhibit the activation of NNK.


Reversal of aflatoxin induced liver damage by turmeric and curcumin. The effect of certain food additives on aflatoxin production by Aspergillus parasiticus has been studied in vitro. Extracts of turmeric (Curcuma longa), garlic (Allium sativum) and asafoetida (Ferula asafoetida) inhibited the aflatoxin production considerably (more than 90%) at concentrations of 5-10 mg/ml. Similar results were also seen using butylated hydroxytoluene, butylated hydroxyanisole and ellagic acid at concentration 0.1 mM. Curcumin, the antioxidant principle from Curcuma longa did not have any effect on aflatoxin production. Turmeric and curcumin were also found to reverse the aflatoxin induced liver damage produced by feeding aflatoxin B1 (AFB1) (5 micrograms/day per 14 days) to ducklings. Fatty changes, necrosis and biliary hyperplasia produced by AFB1 were considerably reversed by these food additives.


Immunological mechanism of action of the tumor reducing peptide from mistletoe extract (NSC 635089) cellular proliferation. A peptide isolated from the Viscum album extract (Iscador) has been reported earlier to the both cytotoxic and tumour reducing. Spleen cells from animals treated with a very small quantity of this peptide were found to have increased response to phytohaemagglutinin and Concanavalin-A, indicating that more mature lymphocytes are produced by the peptide administration. Moreover injection of the peptide at the lesion site produced infiltration of lymphocytes and macrophages and finally producing necrosis of the tumor. This peptide also stimulated macrophages in vitro and in vivo and activated macrophages were found to have cytotoxic activity towards L-929 fibroblasts.


TGF-alpha and IGF-I effects on calcium ion transients in human breast cancer cells. In order to evaluate the potential role of calcium as an intracellular messenger for IGF-I and TGF-alpha action on breast cancer cell proliferation, we determined whether these growth factors induce any change in [Ca2+]i using fura-2 loaded cells. The hormone independent BT-20 and MDA-MB-231 cells were refractory to the mitogenic actions of exogenously added IGF-I and TGF-alpha. TGF-alpha administration, however, stimulated [Ca2+]i transients in the BT-20 cells. IGF-I and TGF-alpha stimulated DNA synthesis in the MCF-7 and T47D cells. These growth factors did not, however, stimulate any changes in [Ca2+]i in these cells. These data support the idea that receptor-mediated phospholipid hydrolysis does not serve a major signalling function for driving human breast cancer cells into DNA synthesis.


Differential inhibition by staurosporine of phorbol ester, bryostatin and okadaic acid effects on mouse skin. The tumor promoters 12-O-tetradecanoyl-phorbol-13-acetate (TPA), a strong activator of protein kinase C (PKC) and okadaic acid, which is ineffective in this respect, induce a rapidly developing ('early') edema of the mouse ear. Bryostatin, another potent activator of PKC, is unable to induce an 'early' edema but causes a more delayed development of edema at a time when most of the PKC is down-regulated. The PKC inhibitor staurosporine neither inhibits the early TPA- nor the late bryostatin-induced edema, but suppresses the okadaic acid-induced edema very effectively. TPA as well as bryostatin, but not okadaic acid cause a down-regulation of PKC, which is not inhibited by staurosporine. The calmodulin antagonist cyclosporine A, which does not suppress PKC activity, very effectively inhibits the TPA-induced edema and down regulation of PKC. Hence we conclude that protein phosphorylation catalyzed by staurosporine-suppressable PKC is not involved in the induction of edema and PKC down-regulation by TPA but that a calmodulin dependent process may play a critical role in these and other TPA effects in mouse skin.


Promoting effects of 6-mercaptopurine on carcinogenesis in various organs of F344 rats. Possible promoting effects of 6-mercaptopurine (6-MP) on carcinogenesis in various organs, including the hematopoietic system, were investigated in female F344 rats, using a 2-stage carcinogenesis model. 6-MP was given as a dietary supplement (50 ppm) for 35 weeks subsequent to wide-spectrum initiation with N-ethyl-N-nitrosourea (ENU). Various tumors were observed in the carcinogen-initiated groups. No significant influence of 6-MP on their development, including the occurrence of leukemia, was apparent. However, the incidences of some proliferative lesions in the lung, intestine and kidney were slightly higher in the ENU/6-MP group than the ENU group. Further studies may be needed on promoting effects of 6-MP, based on dose-effect relation using several 6-MP doses and/or other initiators.


Inhibitory effect of 5-hydroxy-4-(2-phenyl-(E)-ethenyl)-2(5H)-furanone, a novel synthesized retinoid, on azoxymethane-induced intestinal carcinogenesis in rats. Modifying effects of 5-hydroxy-4-(2-phenyl-(E)-ethenyl)-2(5H)-furanone, a novel synthesized retinoid (KYN-54), on intestinal carcinogenesis were examined in a rat model using azoxymethane (AOM). A total of ninety male F344 rats, 6 weeks old, were divided into 4 groups. Group 1 (20 rats) was fed a diet containing KYN-54 at a concentration of 0.02% for 3 weeks, during which time 2 s.c. injections of azoxymethane (15 mg/kg) were applied and then kept on a basal diet until the end of the experiment (1 year). Group 2 (30 rats) was given azoxymethane as in group 1 and fed the basal diet throughout, without synthetic retinoid exposure. Group 3 (20 rats) was administered KYN-54 at the commencement of the experiment, but not given the carcinogen. Group 4 (20 rats) received a basal diet alone throughout the experiment and served as a control. Intestinal tumors were seen in groups 1 and 2, their incidence and average number in group 1 (74%, 1.07 +/- 0.87) being significantly less than in group 2 (39%, 0.56 +/- 0.78) (P < 0.02 and P < 0.05, respectively). These results suggest that the synthetic retinoid might be a promising chemopreventive agent for intestinal neoplasia.


Microscopic localization of sterically stabilized liposomes in colon carcinoma-bearing mice. Using light and electron microscopy, we investigated the in vivo distribution of liposomes sterically stabilized by specific lipids which prolong their circulation in blood. Tissue distribution of sterically stabilized liposomes composed of distearoyl phosphatidylcholine:cholesterol:monosialoganglioside GM1 (10:5:1)-encapsulated 67Ga-Desferal indicates that more than 30% of liposomes still remain in the blood at 24 h after tail vein injection. Moreover, such liposomes accumulated in tumors (C-26 colon carcinoma cells implanted s.c.), reaching almost the same level of uptake as liver (approximately 20% injected dose/g tissue). The microscopic localization of liposomes labeled with encapsulated colloidal gold or rhodamine-labeled dextran coincided well with the tissue distribution. To evaluate circulation parameters, two sizes of gold-containing egg phosphatidylcholine:cholesterol:distearoyl phosphatidylethanolamine (derivatized at its amino position with a 1900 molecular weight segment of polyethylene glycol) (10:5:0.8) liposomes were injected. The plasma was examined by electron microscopy of negative-stained preparations at 0.5, 4, and 24 h after liposome injection. It was found that the ratio of small (less than 100 nm diameter) to large (greater than 100 nm) liposomes increased with time, indicating a much faster clearance of the larger liposomes. To detect the localization of liposomes in various tissues, appropriate samples were fixed 24 h after the injection of gold-containing liposomes (between 80 and 100 nm in diameter) composed of egg phosphatidylcholine:cholesterol:monosialoganglioside GM1 (10:5:1) or egg phosphatidylcholine:cholesterol:derivatized distearoyl phosphatidylethanolamine. The tissues examined for this study included normal liver, bone marrow, and implanted neoplasms. Silver-enhanced colloidal gold was found predominantly within Kupffer cells in the normal liver and within macrophages in the bone marrow. Rarely were any silver-enhanced gold particles detected in hepatocytes. In all preparations, electron microscopy revealed the presence of gold in endosomes and lysosomes of fixed sinusoidal lining macrophages in the liver and bone marrow. Peripheral to the implanted tumors, silver enhancement revealed gold in small blood vessels and focally beyond the vessel boundaries in extracellular spaces around tumor cells. Gold particles were not observed within the tumor cell cytoplasm. At the tumor border, nonenhanced gold was occasionally seen by electron microscopy in cells of the mononuclear phagocyte system. We obtained the same localization pattern as with silver enhancement by using an alternative aqueous content marker, rhodamine B isothiocyanate-dextran. We conclude that liposomes of specific composition, which have the ability to remain in circulation with a half-life of 12-24 h, are also able to transverse the endothelium of small blood vessels, including those in tumors, and extravasate into extracellular spaces.(ABSTRACT TRUNCATED AT 400 WORDS)


Molecular and genetic analysis of liver oncogenesis in transforming growth factor alpha transgenic mice. Overexpression of a transforming growth factor alpha (TGF-alpha) transgene induced the development of liver tumors in 69 of 93 (74%) adult male mice. To identify factors associated with oncogenesis, liver tumors from transgenic animals were characterized at the molecular level. TGF-alpha RNA transcripts were elevated in 17 of 25 (68%) liver tumors, relative to adjacent nontumorous tissue. Expression of the endogenous c-myc and insulin-like growth factor II genes was enhanced in 7 of 19 (37%) and 12 of 16 (75%) tumors, respectively. In contrast, epidermal growth factor receptor RNA levels were unchanged or reduced in all liver tumors, and mutations were not detected in either the Ha-ras or Ki-ras genes. The occurrence of liver tumors in castrated TGF-alpha transgenic mice was reduced about 7-fold, while in ovariectomized transgenic animals the incidence was increased about 6-fold. The progeny of a cross between CD1-derived TGF-alpha transgenic (MT42) and C57BL/6 mice exhibited no reduction in tumor burden (83%); however, the incidence of tumor formation in MT42 x FVB/N offspring was substantially lower (19%). We conclude that in these transgenic mice TGF-alpha promotes tumor formation and appears to play a major role in tumor progression. Moreover, other factors that may collaborate in TGF-alpha-induced hepatocarcinogenesis include c-myc, insulin-like growth factor II, sex hormones, and the genetic background upon which the transgene operates.


Cathepsin D as a prognostic indicator for node-negative breast cancer patients using both immunoassays and enzymatic assays. This is a retrospective study on 162 node-negative patients, with both biochemical and clinical factors being measured for determination of prognostic markers. Steroid receptors were measured on all tumors, while tumor size, histological grade, ploidy status, and cell cycle kinetics indicators could not be found or measured on 25 or less of the patient group. The primary focus of this study was the measurement of cathepsin D, analyzed by two different procedures, and 161 of the 162 patients had at least one value. The antigenic assay was performed using the US-CIS kit, and it was sensitive and reproducible. A biochemical assay using the enzymatic activity of cathepsin D was developed, and it gave proportional values, compared to the antigenic assay values (r2 = 0.79). Our results indicated that the mean antigenic levels were 20% higher than the biochemical assay levels (P = 0.001). High levels of cathepsin D by the antigenic assay predicted poor relapse-free (P = 0.0001) and overall (P = 0.0004) survival. High levels of cathepsin D by the biochemical assay also predicted poor relapse-free (P = 0.031) and overall (P = 0.0013) survival. The cathepsin D values were still useful as predictors of outcome after multivariate analysis. Several other factors, such as grade and S phase, were useful as additional prognostic indicators. In conclusion, cathepsin D is the most useful marker in node-negative patients, and the analysis can be performed by both a biochemical and an antigenic assay.


Quantitative imaging of a radiotherapeutic drug, Na2B12H11SH, at subcellular resolution in tissue cultures using ion microscopy. The effectiveness of boron neutron capture therapy is predicted to be dependent not only on the amount of boron taken up by the target cells but also on the intracellular distribution of boron. Using the isotopic imaging technique ion microscopy, we have quantitatively determined uptake and intracellular distribution of Na2B12H11SH, a promising boron drug for boron neutron capture therapy, in four human cell lines: U87 glioblastoma cells, HeLa epithelioid carcinoma cells, GM 2408b mutant skin fibroblasts, and GM 3348b skin fibroblasts. The boron uptake of all four cell lines, after exposure to 100-500 micrograms/ml Na2B12H11SH, increased as the dosages were increased but showed a tendency toward saturation. Boron was more concentrated in the cytoplasm than in the nucleus but was not strongly localized within cells. There were no significant differences in boron uptake among the four cell lines. A retention experiment identified at least two different intracellular boron pools, and cells lost greater than 60% of intracellular boron within 1 h upon changing to Na2B12H11SH-free medium, indicating a largely low affinity binding.


Expression and functional role of the p75 interleukin 2 receptor chain on leukemic hairy cells. Hairy cell leukemia is a chronic lymphoproliferative disorder characterized by the expansion of neoplastic B-cells expressing the p55 chain of the interleukin 2 receptor (IL-2R) system that is recognized by anti-CD25 monoclonal antibodies (mAb) and binds interleukin 2 (IL-2) with low affinity. In the present study we investigated leukemic hairy cells (HC) for the presence of the p75 IL-2R chain which binds IL-2 with intermediate affinity and plays a crucial role in transducing the message to the cell. For this purpose, we tested highly enriched leukemic HC from six hairy cell leukemia patients for the presence of IL-2R transcripts and for the expression of the p55 and p75 IL-2R chains on their surface membrane by flow cytometry and immunoprecipitation analyses. The functional role of IL-2 in the regulation of HC proliferation was also investigated. Our results indicate that freshly isolated HC express detectable messages for both the p75 IL-2R and the p55 IL-2R. Flow cytometry analysis demonstrated detectable levels of p75 IL-2R on the HC from all patients tested. A mixture of two specific mAb was able to immunoprecipitate detectable amounts of p75 IL-2R from leukemic HC. When leukemic HC were cultured in the presence of several concentrations of IL-2 a low proliferative response was observed. Moreover, the IL-2-driven proliferation of HC was markedly inhibited by anti-p75 IL-2R mAb and to a lesser extent by anti-p55 IL-2R mAb. These findings provide direct evidence of the expression of different IL-2 receptors on leukemic HC and suggest that these molecules might play a role in leukemic cell growth.


Amplification of genes within the chromosome 11q13 region is indicative of poor prognosis in patients with operable breast cancer. Amplification of the chromosome 11q13 region, which harbors the BCL1 region and the PRAD1, EMS1, HSTF1, and INT2 genes, was found in 36 (16%) of a series of 226 breast carcinomas. In the 153 patients with stage I-IIIa disease who had received no therapy prior to surgery and who were treated with curative intent, 11q13 amplification was associated with the presence of lymph node metastases (P less than 0.002). The presence of an 11q13 amplification was associated with a significantly shorter relapse-free survival (P less than 0.002) and a higher breast cancer-specific mortality (P less than 0.003). Stepwise multivariate analysis showed that, in addition to lymph node status, 11q13 amplification was the best predictor for short survival. Stratified log-rank analysis indicated that, within the group of lymph node-positive breast cancer patients, 11q13 amplification identifies a subgroup at high risk.


Tumor progression in vivo: increased soybean agglutinin lectin binding, N-acetylgalactosamine-specific lectin expression, and liver metastasis potential. Tumors which grew out from threshold s.c. inocula of L5178Y-F9 and SL2-5 murine T-cell lymphomas in syngeneic DBA/2 mice exhibited a unified natural defense-resistant phenotype including an increased tumorigenicity and correlating reductions in susceptibility to natural antibodies, natural killer cells, and activated macrophages in vitro. The metastatic potential and cell surface saccharide expression of these cells were determined to assess the impact of growth from a small tumor focus in vivo on subsequent metastatic ability and to determine whether there was any association with changes in cell surface carbohydrates, which have been implicated now for many years in tumor development. A significantly increased liver-colonizing ability was observed following i.v. injection. The most consistent change in cell surface saccharide expression detected in studies using five lectins was an increase in N-acetyl-D-galactosamine (D-GalNAc)-specific soybean agglutinin (SBA) binding. The log of experimental liver metastasis, SBA binding, and the percentage of hepatocyte rosetting of the parental and in vivo-selected cells exhibited significant direct correlations. While inhibition of rosetting with in vivo-selected lines by D-GalNAc and galactose was consistent with the involvement of the D-galactose/D-GalNAc-specific hepatocyte receptor, preincubation of the tumor cells but not hepatocytes with D-GalNAc inhibited hepatocyte rosetting and D-GalNAc inhibited homotypic tumor cell binding. These data suggest a role for a saccharide-specific, lectin-like receptor on tumor cells in both interactions and therefore in the increased experimental liver metastasis. Furthermore, the increased expression of D-GalNAc-inhibitable SBA binding sites on the in vivo-selected variants should increase the homotypic binding by the D-GalNAc-specific lectin-like receptors on the tumor cells providing a rationale for the direct relationship observed between increased SBA binding and i.v. metastatic potential.


Homogeneously staining region in anthracycline-resistant HL-60/AR cells not associated with MDR1 amplification. Anthracycline-resistant HL-60/AR cells and their drug-sensitive HL-60/S counterparts were characterized by karyotypic analysis and examined for the overexpression of DNA and mRNA sequences coding for P-glycoprotein (Pgp). The HL-60/S cells were karyotypically stable over a 5-year period of study (1986-1991), except for an additional small Giemsa-positive band noted at 7q22 in cultures harvested in 1987, but not in 1986. This change did not affect drug sensitivity. The drug-resistant HL-60/AR cells examined in 1986, 1987, and 1991 demonstrated a very stable karyotype. The most striking feature was a large homogeneously staining region in the long arm of chromosome 7 (7q11.2), and translocation of the remainder of the long arm to another centromere. Other changes in the HL-60/AR cells included inversion in 9q, partial deletion of the short arm of chromosome 10p, addition of material to the p arm of der(16), loss of chromosome 22, and the appearance of a new marker chromosome. Both HL-60/S and the HL-60/AR cells were found not to amplify DNA or mRNA sequences coding for the Pgp. Thus, although the HL-60/AR cells possess the classical multidrug resistance phenotype and demonstrate a homogeneously staining region near the region of the MDR1 gene, their resistance is due to mechanisms other than those coded for by MDR1.


Antisense-mediated specific inhibition of P120 protein expression prevents G1- to S-phase transition. A pentadecadeoxyribonucleotide (5'-AAAGCCCCCCACCAC), complementary to a splice junction site of mRNA for human proliferation-associated nucleolar protein P120, inhibited expression of the P120 gene and the mitogen-induced proliferation of human lymphocytes. The inhibition of P120 gene expression and proliferation was concentration dependent and reached 90% at 200 microM, as measured by [3H]thymidine uptake and by densitometric scanning of Northern (mRNA) and Western (protein) blots of P120. Inhibition was not observed in cells treated with the correspondent nonsense oligomer. P120 antisense oligomer treatment prevented S-phase entry of mitogen-stimulated lymphocytes, as determined by flow cytometric analysis, but did not block G0-G1 transition assessed by morphological blast transformation and induction of [3H]uridine incorporation. Results of this study suggest that P120 expression may be required for the upregulation of nucleolar function necessary for cell proliferation.


Epigenetic silencing of the DNA repair enzyme O6-methylguanine-DNA methyltransferase in Mex- human cells. Regulation of the expression of the DNA repair enzyme O6-methylguanine-DNA methyltransferase (MGMT) has been investigated in a number of human lymphoblastoid cell lines. In a number of Mex- cell lines that do not express methyltransferase activity, CpG sequences in the mgmt gene were hypomethylated with respect to methyltransferase-expressing Mex+ lines. In the cell line GM1953(S), in which the mgmt gene is coregulated with the thymidine kinase and galactokinase genes, reexpression of all three activities was experimentally induced. In this case, the mgmt gene in the nonexpressing cells was found to be hypermethylated and underwent a demethylation at CpG sequences that was coincident with the reappearance of the mgmt mRNA and the three enzyme activities. The simultaneous silencing of three activities in these cells was correlated with an increase in DNA 5-methylcytosine that was widespread throughout the genome. The data indicate that MGMT expression can be controlled epigenetically in human lymphoid cell lines, although the relationship between cytosine methylation and MGMT expression is complex. Furthermore, the rapid alterations in methylation in GM1953(S) cells indicate the existence of signals that can induce widespread and abrupt alterations in cytosine methylation in human cells in culture.


Enhanced sensitivity of human colon tumor cell lines in vitro in response to thermochemoimmunotherapy. We have investigated the cytotoxic responses in vitro of three human colon tumor cell lines with epithelial-like morphology, DLD-1, HCT-15, and HT-29, to thermochemoimmunotherapy with hyperthermia (42 degrees C for 2 h), carboplatin, and recombinant human tumor necrosis factor (TNF). Dose ranges of carboplatin and recombinant human TNF were administered essentially simultaneously and were followed 1 h later by hyperthermia. A two-tiered approach was used to evaluate cytotoxicity. In the first tier, a 5-day microcytotoxicity assay using vital dye staining was done; the effect on surviving fraction of simultaneously varying carboplatin and recombinant human TNF doses was evaluated by response surface methodology. From this analysis doses were selected for use in the second-tier clonogenic survival assays. A similar treatment protocol was used in clonogenic assays. Both assays revealed significant interline treatment response heterogeneity. Only the HCT-15 cells were sensitive to TNF alone; carboplatin activity against all three tumor cell lines was enhanced by TNF. Hyperthermia had minimal effect as a sole agent but enhanced the effects of carboplatin and TNF in DLD-1 and HCT-15 cells. Triple modality treatment resulted in 3-4-log decreased survival and could reduce cytotoxic resistance expressed against single- or dual-modality treatments by some of these cells.


Selenoperoxidase-mediated cytoprotection against merocyanine 540-sensitized photoperoxidation and photokilling of leukemia cells. Photodynamic therapy with the lipophilic sensitizing dye merocyanine 540 (MC540) is a promising new approach for extracorporeal purging of neoplastic cells from autologous remission bone marrow grafts. Resistance-conferring cellular defenses against the cytotoxic effects of MC540/photodynamic therapy have not been well characterized. This study focuses on the cytoprotective effects of the glutathione-dependent selenoperoxidases GPX and PHGPX, which can detoxify a wide variety of hydroperoxides, including lipid-derived species (LOOHs). Murine leukemia L1210 cells were grown in 1% serum media without [L.Se(-)] and with [L.Se(+)] selenium supplementation. L.Se(-) cells expressed 10- to 20-fold lower GPX and PHGPX activities than L.Se(+) controls and were markedly more sensitive to MC540-mediated photoperoxidation (LOOH formation) and clonally assessed photokilling. Susceptibility of L.Se(-) cells to photoperoxidation and photokilling could be fully reversed to L.Se(+) levels by replenishing Se, and partially reversed by treating with Ebselen, a selenoperoxidase mimetic. Altered lipid composition, greater uptake of MC540, and defective catabolism of H2O2 were all ruled out as possible factors in the elevated photosensitivity of L.Se(-) cells. Human leukemia K562 cells (capable of expressing PHGPX but not GPX) exhibited 5- to 10-fold lower PHGPX activity under Se-deficient relative to Se-sufficient conditions. Although MC540 uptake (nmol/mg lipid) by K562 and L1210 cells was essentially the same, the former were more resistant to photoinactivation. However, like murine counterparts, Se-deficient cells were more susceptible to photoperoxidation and photokilling than Se-sufficient controls. These results clearly demonstrate that GPX and/or PHGPX in L1210 cells and PHGPX in K562 cells play an important cytoprotective role during photooxidative stress. Whether membrane damage due to lipid photoperoxidation is causally related to cell death is not certain; however, the parallel effects of Se deficiency on LOOH formation and cell killing are at least consistent with this possibility.


p53 mutations in breast cancer.

p53 mutations in breast cancer. We have identified and analyzed 41 mutations in p53 in sporadic breast tumors from 136 unselected breast cancer patients and estimate that approximately 40% of such tumors contain p53 mutations. The frequency of G-T transversions and the incidence of guanosine mutations in the nontranscribed strand of the p53 gene were found to be higher than expected, and we suggest, therefore, that exogenous carcinogens have an etiological role in sporadic breast cancers. Mutations were recorded in 44 codons of the p53 gene, with no obvious mutational hot-spots, although mutations at codons 175, 194, 273, and 280 accounted for 25% of the changes. One germ-line mutation was found in 136 patients and so we conclude that constitutional mutation of p53 may be an uncommon etiological factor in breast cancer.


Phosphorous metabolite and cell cycle kinetic response of two human squamous cell carcinomas to radiation. Phosphorous metabolism and cell cycle phase kinetics in response to radiation of two perfused human squamous cell carcinoma cell lines, SQ20B (radioresistant) and SQ38 (relatively radiosensitive), embedded in both basement membrane (Matrigel) and agarose gel threads were studied. The findings for these human cancer cells in response to 2- and 50-Gy irradiation are as follows. (a) Well perfused pure cancer cells (both SQ20B and SQ38) in both proliferative (cells embedded in Matrigel) and static (cells embedded in agarose threads) states did not show significant alteration in either phosphorous bioenergetics or membrane metabolites at 24 and 48 h after irradiation, although a large fraction of the population was clonogenically impaired. Previously reported, sensitively detected, metabolite alterations in response to radiation in rodent and human tumors in situ were not seen in these homogeneous cancer cell populations. (b) The radiosensitive squamous cell carcinoma cell lines SQ38 exhibited G1 block (from 54.38 +/- 1.40% in control to 73.93 +/- 1.01% after irradiation; mean +/ SD) in response to low-dose 2-Gy irradiation and G2 block (from 12.98 +/- 2.15% in control to 25.6 +/- 3.15% after irradiation) in response to high-dose 50-Gy irradiation, while the radioresistant cell line SQ20B showed only conventional G2 block in response to both doses. The differential cell cycle phase response may indicate the difference in radioresistance. (c) The membrane metabolites (including phosphomonoesters and phosphodiesters) and phosphocreatine gradually increased from the early passages to late passages, suggesting that cell proliferation rates were increasing as the cells adapted to tissue culture. The results suggest that the radiation-induced metabolite changes observed in solid tumors in situ may not be a direct response to interim changes within the cancer cells but, rather, a consequence of radiation damage either to the vasculature or to other host-mediated factors.


Highly sensitive, specific detection of O6-methylguanine, O4-methylthymine, and O4-ethylthymine by the combination of high-performance liquid chromatography prefractionation, 32P postlabeling, and immunoprecipitation. A highly sensitive and specific method for the detection of O6-methylguanine (O6-meG), O4-methylthymine (O4-meT), and O4-ethylthymine (O4-etT) has been established by combining prefractionation by high-performance liquid chromatography (HPLC), 32P postlabeling, and immunoprecipitation by monoclonal antibodies (PREPI method). DNA was enzymatically hydrolyzed to 2'-deoxynucleoside-3'-monophosphates (3'-dNps). Each alkyl 3'dNp was separated by reverse-phase HPLC, radiolabeled at the 5' position with [gamma-32P]ATP and polynucleotide kinase. After removing 3'-phosphate for better recognition by the antibodies, the resulting alkyl nucleotides were further fractionated by HPLC and finally precipitated specifically with respective antibodies. The detection limits were 1 fmol for all the alkyl nucleotides analyzed, so that one adduct in 10(8) of its normal counterpart nucleotide can be determined using approximately 100 micrograms (O4-meT and O4-etT) or approximately 150 micrograms (O6-meG) of DNA, i.e., 3-5 x 10(7) cells corresponding to approximately 10 ml of peripheral blood or a few hundred milligrams of tissue. By the use of the PREPI method, three leukocyte and three liver DNA samples from Japanese living in the Tokyo area were analyzed with respect to O-alkyl adduct content. O6-meG was detected in all three of the leukocyte samples (O6-meG:G molar ratio, 1.1 x, 0.8 x, and 1.6 x 10(-8) as molar ratios to guanine). Neither O4-meT nor O4-etT was detected (detection limit, O4-alkylT:thymine molar ratio less than 0.5 x 10(-8)). Among the liver samples analyzed, two cases showed positive O6-meG values (4.2 x and 1.1 x 10(-7) O6-meG:guanine molar ratios). Contrary to the leukocyte DNA, O4-meT (3.9 x, 4.3 x, and 7.5 x 10(-8) as O4-meT:thymine) and O4-etT (1.9 x, 4.9 x, and 8.7 x 10(-8) as O4-etT:thymine) were detected in all the liver samples. These results indicate the validity of the PREPI method for molecular epidemiological studies on DNA alkylation products.


Putrescine-dependent invasive capacity of rat ascites hepatoma cells. The effects of inhibitors of polyamine synthesis on the invasive capacity of rat ascites hepatoma (LC-AH) cells were examined by in vitro assay of penetration of the LC-AH cells through a monolayer of calf pulmonary arterial endothelial (CPAE) cells. Pretreatment of LC-AH cells with alpha-difluoromethylornithine (DFMO), an inhibitor of ornithine decarboxylase, before seeding them onto a CPAE cell monolayer and culturing them for 24 h in the absence of DFMO decreased the number of penetrating tumor cells time and dose dependently (about 35% of the maximal inhibition) without affecting their viability or proliferative activity. DFMO treatment caused a marked decrease in the intracellular level of putrescine but not of spermidine or spermine. The DFMO-induced decreases in invasive capacity and putrescine level were almost completely reversed by the addition of putrescine to the medium during pretreatment with DFMO or invasion assay but were not affected by exogenous spermidine or spermine. No change in the invasive capacity was observed when the CPAE cells were treated with DFMO and the LC-AH cells with methylglyoxal-bis(guanylhydrazone), an inhibitor of S-adenosylmethionine decarboxylase, which depressed the spermidine and spermine levels but increased the putrescine level in the LC-AH cells. These results suggest that intracellular putrescine modulates the in vitro invasive capacity of LC-AH cells.


Involvement of the spleen in preleukemic development of a murine retrovirus-induced promonocytic leukemia. An acute myeloid leukemia can result from the inoculation of Moloney murine leukemia virus into BALB/c mice undergoing a 2,6,10,14-tetramethylpentadecane-induced chronic inflammatory response in the peritoneal cavity. This leukemia is ultimately observed in the peritoneal cavity as an ascites with cells infiltrating the granulomatous tissue. It has been proposed, however, that hematopoietic organs such as the spleen and bone marrow are involved in preleukemic development of Moloney murine leukemia. Therefore, to determine if the spleen plays a role in this development, mice were splenectomized at various times relative to virus inoculation. When splenectomies were performed 3 days before and 2, 4, 6, and 8 weeks after virus inoculation there was, in all cases, a decreased death rate compared to sham-splenectomized controls. The greatest difference in death rate due to promonocytic leukemia was observed when mice were splenectomized at 4 weeks after virus inoculation. The decrease in disease incidence observed as a result of splenectomy was not caused by decreased virus spread in hematopoietic organs or an alteration in the profile of the cellular infiltrate in the granuloma. It was found, however, that the spleens of 2,6,10,14-tetramethylpentadecane-treated mice, relative to those of normal mice, have a significantly increased number of granulocyte-macrophage colony-forming cells and a slightly increased number of multipotential colony-forming cells. These observations suggest that a population of target cells for transformation, consisting of granulocyte-macrophage precursor cells, may reside in the spleen. Alternatively, partially transformed cells may reside temporarily in the spleen during the developmental stages of the disease process.


Marked enhancement of rat urinary bladder carcinogenesis by heat-killed Escherichia coli. Chronic urinary tract infection is an important risk factor for the development of carcinoma in the human urinary bladder. To test the effect of chronic persistent inflammation on bladder carcinogenesis, we instilled heat-killed Escherichia coli (1 x 10(8) cells suspended in 0.5 ml of phosphate-buffered 2.1% NaCl solution) twice a week into the heterotopically transplanted rat urinary bladders in which carcinogenesis was initiated by a single dose (0.25 mg) of N-methyl-N-nitrosourea. When compared with the control animals, the rats treated with killed E. coli showed significantly enhanced bladder tumorigenesis, as reflected by an increase in the incidence of tumor (P = 0.05) and a 6- to 40-fold increase in the number of tumors per bladder (P less than 0.0001). The tumors were characterized by intraepithelial clusterings of neutrophils and by chronic inflammation and marked capillary proliferation in the tumor stroma. All of these features were rare in tumors in the control groups. The accelerated cell proliferation induced by killed E. coli treatment appears to play a significant role in the enhancement of tumorigenesis.


Simian virus 40 large T antigen directed by transcriptional elements of the human surfactant protein C gene produces pulmonary adenocarcinomas in transgenic mice. A model of pulmonary adenocarcinomas was produced in transgenic mice harboring a chimeric gene comprising the SV40 large T antigen under the control of a transcriptional region derived from the human surfactant protein C (SP-C) gene. Transgenic mice succumbed with pulmonary tumors within 4-5 months of age. By histology, the tumors were adenocarcinomas with lepidic, papillary, and solid growth patterns that were indistinguishable from adenocarcinomas occurring in humans. Immunocytochemistry demonstrated the lack of staining for neuroendocrine markers, consistent with the identification of the tumors as non-small cell rather than small cell carcinomas. The presence of SV40 large T mRNA in the lung and tumors was detected by in situ hybridization and Northern blot analysis. Exogenous SV40 large T mRNA and endogenous CC10 (a nonciliated respiratory epithelial cell marker) and SP-C (a Type II alveolar cell marker) mRNAs were expressed at variable levels in the lung tumors. SV40 large T mRNA and CC10 mRNA were detected in the majority of tumors, while SP-C mRNA was detected less frequently. The heterogeneity of bronchiolar and alveolar cell markers in the tumors from the transgenic mice supports the concept that tumorigenesis was initiated in distinct subsets of epithelial cells that produce characteristic adenocarcinomas of the lung.


Monovalent immunotoxin containing truncated form of Pseudomonas exotoxin as potent antitumor agent. Recombinant truncated forms of Pseudomonas exotoxin A that lack the cell binding domain of Pseudomonas exotoxin A were coupled to an F(ab') fragment of a monoclonal antibody HB21 directed against the human transferrin receptor. One of these was NlysPE40. The other, NlysPE38QQR, has two amino groups on residues near the NH2-terminus and has no amino groups near the COOH-terminus. The proteins were linked by a stable thioether bond that connected the sulfhydryl group present in the hinge region of the antibody fragment to an amino group on the toxin. The F(ab')-PE40 immunotoxin, containing NlysPE40, exhibited potent cytotoxic activity on human carcinoma cell lines with a concentration of immunotoxin at which isotope incorporation falls by 50% when compared to nontreated cells (ID50) of 5.3 pM (0.5 ng/ml) on both the epidermoid carcinoma A431 and on the colon carcinoma Colo205. Immunotoxins made with whole antibody were considerably less active, with an ID50 of 15.9 pM (3.1 ng/ml) on these cell lines. F(ab')-PE38QQR, the immunotoxin containing NlysPE38QQR, was found to be the most active agent with an ID50 of 1.05 pM (0.1 ng/ml) on A431 cells. The greater cytotoxicity of immunotoxins containing fragmented antibody was probably due to the higher binding affinity of F(ab') conjugates in comparison to whole antibody conjugates to the transferrin receptor. The increase in cytotoxic activity of the immunotoxin made with NlysPE38QQR than that with NlysPE40 may reflect selective coupling of the toxin through NH2-terminal amino groups. The monovalent and divalent immunotoxins had dose-dependent antitumor effects on human epidermoid carcinoma xenografts in nude mice. A431 tumors completely regressed in all animals at a total dose of 105 pmol (10 micrograms) of F(ab')-PE38QQR and of 154 pmol (30 micrograms) of IgG-PE38QQR. Furthermore, the F(ab') immunotoxin was less toxic to mice than the conjugate containing IgG (840 pmol or 80 micrograms of total dose causing measurable adverse effects versus 208 pmol or 40 micrograms, respectively). Thus, a truncated Pseudomonas exotoxin A molecule coupled to the F(ab') fragment of an antibody is more active and less toxic in mice than an immunotoxin made with a whole antibody. Therefore, the therapeutic index for the monovalent immunotoxin is about four times better than that for the divalent immunotoxin.


Differential inhibition of the epidermal growth factor-, platelet-derived growth factor-, and protein kinase C-mediated signal transduction pathways by the staurosporine derivative CGP 41251. The microbial alkaloid staurosporine is a potent but nonselective inhibitor of protein kinases. The derivative CGP 41251 has been shown to exert a high degree of selectivity for inhibition of protein kinase C activity. Both compounds are powerful inhibitors of proliferation of both normal and transformed cells in vitro and exert antitumor efficacy in vivo. In this work we have studied the mode of action of these compounds by analyzing their effects on early events in the induction of proliferation by different growth stimuli. Both drugs blocked the phorbol ester-induced expression of the c-fos proto-oncogene. The effect of CGP 41251 was reversible, since its removal led to a normal expression of c-fos mRNA in response to phorbol 12-myristate 13-acetate. Submicromolar concentrations of CGP 41251 and staurosporine directly inhibited both the platelet-derived growth factor (PDGF) receptor autophosphorylation and the c-fos mRNA expression induced by PDGF stimulation of intact BALB/c 3T3 cells. In contrast, ligand-induced epidermal growth factor receptor autokinase activity in A431 carcinoma cells and epidermal growth factor-dependent c-fos mRNA expression were relatively insensitive to inhibition by CGP 41251. Staurosporine suppressed signal generation by the epidermal growth factor receptor by reducing overall levels of the receptor. We conclude that CGP 41251 is a potent reversible inhibitor of protein kinase C and PDGF-mediated signal transduction. It inhibits the kinase activity of both protein kinase C and the PDGF receptor tyrosine kinase and the subsequent signaling cascade. The broad inhibition of kinases by staurosporine is also reflected at the cellular level and might contribute to the high toxicity of this compound, in comparison to CGP 41251.


Effects of tamoxifen adjuvant therapy and a low-fat diet on serum binding proteins and estradiol bioavailability in postmenopausal breast cancer patients. Serum was collected at intervals from postmenopausal breast cancer patients to determine the effects of tamoxifen adjuvant therapy and a low-fat dietary intervention, alone and in combination, on sex hormone-binding globulin (SHBG) concentrations and circulating estradiol bioavailability. Serum corticosteroid-binding globulin and follicle-stimulating hormone were also assayed as indicators of patient compliance to tamoxifen therapy. The immunoreactive SHBG concentration was higher (P less than 0.001) in 22 patients who had been treated with tamoxifen for 6-36 weeks when first sampled, compared with 27 who were not receiving tamoxifen therapy. Tamoxifen also produced a reduction in the percentage non-protein-bound estradiol (P less than 0.001) and percentage albumin-bound estradiol (P less than 0.01), the two biologically available fractions, and a corresponding increase in the percentage SHBG-bound estradiol (P less than 0.01). A longitudinal study of 7 patients showed significant reductions in the percentage of albumin-bound estradiol and an increased percentage of SHBG-bound estradiol, after 3-6 months of tamoxifen; after 12-18 months there was also a significant decrease in the non-protein-bound estradiol fraction. We conclude that in postmenopausal breast cancer patients the redistribution of circulating estradiol, with reduced bioavailability, provides an additional mechanism to those demonstrated previously for the therapeutic activity of tamoxifen. Another 12 patients receiving tamoxifen and 8 who were not were followed for 6-12 months on a low-fat diet (fat comprised 20% of the total calories). The dietary intervention had no effect on the serum SHBG concentration or the estradiol distribution. Although tamoxifen increased the serum corticosteroid-binding globulin and partially suppressed the follicle-stimulating hormone concentrations, the responses obtained were less consistent compared with those of the SHBG levels.


Postvaccinal sarcomas in the cat: epidemiology and electron probe microanalytical identification of aluminum. An increase in fibrosarcomas in a biopsy population of cats in the Pennsylvania area appears to be related to the increased vaccination of cats following enactment of a mandatory rabies vaccination law. The majority of fibrosarcomas arose in sites routinely used by veterinarians for vaccination, and 42 of 198 tumors were surrounded by lymphocytes and macrophages containing foreign material identical to that previously described in postvaccinal inflammatory injection site reactions. Some of the vaccines used have aluminum-based adjuvants, and macrophages surrounding three tumors contained aluminum oxide identified by electron probe microanalysis and imaged by energy-filtered electron microscopy. Persistence of inflammatory and immunological reactions associated with aluminum may predispose the cat to a derangement of its fibrous connective tissue repair response, leading to neoplasia.