A study of p53 protein, proliferating cell nuclear antigen, and p21 in Hodgkin's disease at presentation and relapse.

About one fourth of patients with Hodgkin's disease relapse after therapy. The mechanisms that lead to resistance to treatment in these patients are poorly understood. The authors describe the differential protein expression of p53, proliferating cell nuclear antigen (PCNA), and p21 at initial presentation and relapse, and discuss their role in disease progression and resistance to therapy. Thirty-four patients with Hodgkin's disease who had relapsed after standard chemotherapy and radiotherapy regimens were assessed for the expression of p53 protein, PCNA, and p21 protein (waf/cip 1). In 14 of these cases, sequential biopsies performed both at presentation and at relapse were available for the study. Seventy-five percent of the cases were positive for the p53 protein. Tumors at relapse had higher p53 and PCNA scores than those at initial presentation. In the paired samples, a significant increase was noted in the number of p53 and PCNA-positive cells and in the intensity of staining with p53 antibody. Six of seven paired samples tested for p21 showed an increased p21 expression at relapse. These results suggest that, at relapse, Reed-Sternberg (RS) cells and their variants positive for p53, PCNA, and p21 are increased in number and individually have an increased expression of p53, PCNA, and p21 proteins. These findings suggest that therapy failure and relapse may, at least in part, be associated with altered p53, p21, and PCNA pathways. HUM PATHOL 28:549-555. This work was carried out during an exchange fellowship program at the National Cancer Institute, Bethesda. There are no restrictions on its use

The perinatal autopsy: a neglected source of discovery.

The perinatal autopsy is frequently viewed by pathologists as being scientifically unrewarding and contributing little to patient care. To emphasize its importance in patient management, genetic counselling and specific research, a ten-year review (1979-88) was made of the perinatal autopsy experience at Loyola University Medical Center. The 657 deaths included late fetal deaths (22%), early neonatal deaths (51%), late neonatal deaths (10%) and deaths between 29 days and one year (17%). Comparison of the principal causes of death in various groups categorized by birth weight and age revealed significant trends. There has been a steady decline in mortality from immediate complications of immaturity, while the mortality rate from long-term complications of immaturity has not increased. There were increased frequencies of congenital diaphragmatic hernia and cardiac malformations, while the frequency of renal malformations decreased; the etiological significance of these changes requires further evaluation. A correlation of clinical observations with post-mortem findings indicates that newer diagnostic procedures, such as ultrasound, echocardiography and cardiac catheterization, are of limited value for accurate diagnosis of complex anomalies, cystic renal disease and chromosomal anomalies. A substudy included cases up to 18 years of age. The frequency of childhood neoplasia was low (17 cases), and among these cases there was a predominance of haematological malignancies (11 cases). Even with these small numbers, a shift in cause of death from disseminated malignancy to overwhelming infection was apparent. This paper includes the essential elements of a protocol for perinatal autopsies, with illustrations of specific applications. The perinatal autopsy is clearly an undervalued source of information and discovery. Little or no information is available from developing countries, where autopsies could provide information on causes of paediatric mortality and permit recognition of disease patterns, which is so critical to the planning of health services.

Contribution of post-mortem examination to cancer registries.

In developed countries, the post-mortem examination may not be critical to the identification of patients with cancer, since diagnostic sophistication and histological verification are readily available. The autopsy should be considered of increasing importance, however, to epidemiologists and other investigators who use the data generated by cancer registries. The contribution of post-mortem examinations to etiological studies, to documentation of the natural history of disease in a changing environment and to evaluating diagnostic and therapeutic modalities should, in fact, be recognized as essential to the practice of modern medicine and to research. In the less developed countries, cancer registration must depend to a much greater degree on necropsy for identification of cancer patients, since surgical biopsy may be limited or selected and often specimens are not even submitted for examination. The primary site and histology of a tumour are frequently not available from clinical records, which may carry significant inaccuracies. Most of the important observations and contributions relating to the geographical pathology of cancer have made substantial use of post-mortem material. All of the investigative approaches that are dependent on reliable cancer registry data and that involve use of information derived from autopsies are equally applicable to developed and less developed countries. Such information is of enormous importance to public health planning, and the potential for disease prevention cannot be overestimated.

Burkitt's lymphoma cell lines reveal different degrees of tumorigenicity in nude mice.

A quantitative in vivo assay for evaluating the tumorigenicity of Burkitt's lymphoma (BL) cell lines in nude mice is described. It is based on the dose-response kinetics of BL cell lines in pre-irradiated (480 rad) nude mice following the s.c. injection of 4 different cell doses. This model system was used to estimate the xenografting potential of 26 BL cell lines derived from BL patients of different geographic and ethnic origins, as well as lymphoblastoid cell lines (LCLs) established by Epstein-Barr virus (EBV) immortalization of normal B lymphocytes. The results indicate that most BL cell lines are tumorigenic, but LCLs fail to produce progressively growing tumours in nude mice. However, BL cell lines revealed individual degrees of tumorigenicity and accordingly could be divided into 4 groups with high, moderate, low or no tumorigenicity. Preliminary attempts to correlate the xenografting phenotype of BL lines with other characteristics indicate that: (1) the aberrations of chromosome 1 are more often encountered in cell lines with high and moderate tumorigenicity; (2) EBV-positive BL lines do not reveal a higher tumorigenic phenotype in comparison with EBV-negative ones; (3) cell lines carrying translocations t(8;22) and t(2;8) might fall more frequently in the group of lines with high and moderate tumorigenicity; and (4) when LCLs grow in nu/nu mice, rejection always occurs and is associated with massive tumour necrosis. These findings suggest that the tumorigenicity of BL cell lines in immunosuppressed animals is not related with EBV, but with certain chromosomal abnormalities (BL-specific and non-specific) indicating that this in vivo model system can be instrumental for the identification of other factors or stages involved in BL development.

Malignant lymphoma in African children: three decades of discovery.

The recognition of Burkitt lymphoma (BL) as a clinical syndrome and a pathological entity in African children resulted from astute clinical observations (bedside epidemiology), the availability of cancer registry data and accurate pathological interpretation. Following the early studies in Africa, it soon became evident that this tumor occurred worldwide and the excess of cases in Africa was an incidence phenomenon associated with specific environmental factors. The sentinel discovery of the Epstein Barr virus (EBV) and its association with BL stimulated a wide variety of scientific investigations which have had an impact of virtually every discipline and biology. Epidemiological observations linked to modern laboratory techniques have provided etiological insights which implicate specific environmental factors and genetic events in the pathogenesis of BL and other immunoproliferative diseases. Early infection with EBV and holoendemic malaria are clearly of paramount importance in the development of endemic BL (eBL). These factors do not play a role in the majority of sporadic BL (sBL) cases, but immunosuppression and T-cell deregulation almost certainly are common denominators. The final or principle genetic event in both instances would appear to be the chromosome 8 translocation involving the c-myc oncogene and structural alteration. It is expected that the BL model will continue to be a useful one for identifying basic mechanisms in carcinogenesis which may be applicable as well to a variety of non-neoplastic diseases.

Haematological cell proliferation and differentiation responses to perturbations of polyamine biosynthesis.

Combined administration of methylglyoxal-bis-guanylhydrazone (MGBG) (25 mg/kg) with difluoromethylornithine (DFMO), or MGBG alone at a higher dose (50 mg/kg), to mice resulted in a decreased white cell count (WBC) in the peripheral blood while DFMO or MGBG alone at a lower dose (25 mg/kg) had no effect. As expected, DFMO alone increased the number of colony forming units spleen (CFU-s), colony forming units diffusion chamber granulocyte (CFU-dg) and colony forming units culture (CFU-c) in the bone marrow. MGBG treatment led to an increase in CFU-dg alone. Combined treatment seemingly had no effect on marrow stem cells. Total tibial and differential counts were not affected by any of the treatments. Cell proliferation in diffusion chamber cultures, as judged by CFU-dg colony formation, was impaired by MGBG alone or in combination with DFMO, at dose levels which had no effect or increased the precursor cell number in the bone marrow. This effect was partially reversed with either putrescine or spermidine. Determination of intracellular polyamine concentrations, demonstrated decreased putrescine and spermidine levels after DFMO administration. As expected, MGBG treatment resulted in decreased spermidine and spermine levels, concomitant with an increase in putrescine. In mice which received both agents, rather than only MGBG, after 3 days higher intracellular polyamine concentrations were observed. After 11 days, however, there was no significant difference between the two groups.

The gross pathology and histological features of tumours produced by inoculation of human cell lines into immune-deprived mice.

Tumours were raised in both congenitally athymic ('nude') Swiss mice and in neonatally thymectomized, Ara-C-protected, whole-body irradiated CBA mice by subcutaneous inoculation of cells from a variety of cultured human lines. In both types of animal, tumours tended to grow massively at the site of inoculation, with some infiltration of adjacent tissues but only rarely with evidence of metastatic spread. Tumours derived from Burkitt's lymphoma (BL) lines or from EB virus-transformed lymphoblastoid cell lines (LCL) were all classified as high grade malignant lymphomas with a limited range of appearances on conventional histological examination. In the material studied there were no consistent features distinguishing BL-derived from LCL-derived tumours. Cell lines originating from other haematopoietic malignancies tended to produce tumours interpreted as immunoblastic lymphomas though there were distinctive characteristics in some cases, such as highly convoluted or pleomorphic nuclei in the cells of some tumours derived from T-cell leukaemia lines and plasmacytoid differentiation in tumours originating from myeloma lines. Malignant cell lines of epithelial origin gave rise to tumours with the histological appearances of anaplastic carcinomas readily distinguishable from the high grade lymphomas produced by haematopoietic cells.

Gamma heavy chain disease: report of a case associated with trisomy of chromosome 7.

A case of gamma heavy chain disease is reported in a 52-year-old white male who presented with fever and generalized lymphadenopathy. A lymph node biopsy showed malignant lymphoma. A partial transient response was obtained with cyclophosphamide, vincristine, prednisone, and doxorubicin. He died 3 months after diagnosis from disease progression and infectious complications. Chromosome analysis of cells from an involved lymph node showed the presence of trisomy 7. Chromosome abnormalities have been reported in three of ten previously published cases of gamma heavy chain disease. Trisomy of chromosome #7 has not previously been reported.

Treatment of advanced esthesioneuroblastoma with high-dose chemotherapy and autologous bone marrow transplantation. A case report.

A 46-year-old woman presented with an advanced unresectable esthesioneuroblastoma which failed to respond to radiation therapy and one course of chemotherapy. She underwent treatment with high-dose chemotherapy (cyclophosphamide, doxorubicin, and vinblastine) followed by autologous bone marrow transplantation. The major toxicity from the regimen was severe oropharyngeal mucositis. A complete remission was achieved and the patient is free of disease and asymptomatic 3.5 years after treatment.

Opportunities for study of lymphoid neoplasia in Africa.

Animal models have strongly suggested that lymphoid leukaemias and malignant lymphomas might have a viral etiology. Following the first report by Ludwig Gross in 1951 of the transmission of mouse leukaemia by cell-free filtrates inoculated into newborn mice and the subsequent identification of many tumour-inducing type-C viruses in animals, attention was focused on the RNA retroviruses. However, it was work in Africa which led to the recognition of Burkitt's lymphoma as an entity and to the discovery of the Epstein-Barr virus. Many refer to this DNA virus as the first recognized human tumour virus. The research that these early studies stimulated has resulted in enormous progress in our understanding of human carcinogenesis and in highly significant advances in therapy. More recently, the first human retrovirus was identified and characterized in the laboratory of Dr Robert Gallo. There is increasing evidence that this virus, isolated from a patient with a T-cell lymphoma, is etiologically associated with a specific type of T-cell neoplasm first described in Japan. Africa may be considered the cradle of geographical pathology and the potential for generating unique information on cancer etiology and cancer control is still enormous. Lymphoid neoplasms are excellent models for the study of human cancer, particularly in Africa, since as tumours of the immune system, their induction and evolution is strongly influenced by those environmental factors, such as infection and nutrition, which have a profound effect on the immune response. In addition, there is good evidence that at least some of the tumours in this group are associated with specific transforming viruses. There is an immediate need to collect accurate, reproducible and comparable data on the incidence and characteristics of the different types of lymphoid neoplasia and on the populations in which they occur in the various African countries. Such data will provide the basis for undertaking simultaneous or subsequent etiological and therapeutic studies. Realistic approaches to these goals are considered and specific studies relating to questions posed by available information discussed.

Histopathological features of rapidly progressing breast carcinoma in Tunisia: a study of 94 cases.

The histological features of 94 cases of carcinoma of the breast seen in Tunisia were recorded and subsequently correlated with the clinical classification of the patients in terms of poussée évolutive (PEV) categories. Histological features analyzed in the breast tissues and skin included tumor type, nuclear grade, number of mitoses, involvement of the dermis, cutaneous inflammatory infiltrate and edema. Twenty-eight percent of the cases in which the skin was examined showed tumor emboli in the lymphatics of the dermis. The frequency of cutaneous permeation correlated with the PEV categories. The percentage of cases with nuclear grade 3 was higher for the group with rapid progression of the disease (PEV 1,2,3) than for the cases belonging to the PEV-0 category (90% versus 64.2%). The present study demonstrates that a large proportion of breast carcinoma patients in Tunisia suffer from "inflammatory carcinoma of the breast".

Spontaneous lymphosarcoma arising in a nude mouse: characterization in vivo and in vitro.

A lymphosarcoma spontaneously arising in a nude mouse and a continuous cell line (NML-1) derived from it are described and compared. The primary tumor and a transplantable tumor line from it were composed of lymphoid cells, with no C-type viral particles seen by electron microscopy. The culture line was composed of cells with morphologic and functional properties of macrophages; budding C-type particles were abundant. The cells in the tumors produced in nude mice by injection of the NML-1 cells also resembled macrophages morphologically rather than lymphocytes; however, by electron microscopy, no C-type particles were seen. The findings suggest some type of in vivo suppression of complete expression of the virus.