A 58-year old man was admitted because of general malaise in April 1987. Physical examination revealed systemic lymphadenopathy and hepatosplenomegaly. The white blood cell count was 252, 900/microliters with 82% of blasts. Bone marrow aspiration contained 93.8% lymphoblasts, which were positive for TdT and negative for peroxidase reaction. Immunologic marker studies showed OKT 11 positive and CALLA negative. Cytogenetic analysis revealed a clone with 46, XY, t (9; 22) (q34; q11), del(5) (q15) in 12 of the 13 metaphases. Ph1 positive T-acute lymphoblastic leukemia was considered. After AdVP and following AdVEMP (induction) chemotherapy, complete remission was obtained in August 1987. Cytogenetic study at the remission stage showed complete disappearance of Ph1 positive clone. Treatment with BH-AC DMP protocol at the time of recurrence in November 1987, brought no improvement and he died of respiratory failure. Chromosome study at recurrence showed an additional complex abnormal karyotype (double Ph1, +2, 5q-, -10, -13, -17). DNA analysis revealed rearrangements of bcr gene with deletion of 5' side and of TCR delta gene, without any rearrangements in other immunoglobulin genes. From cytogenetic, immunophenotypic and genetic analysis the patient was diagnosed as having acute lymphocytic leukemia (FAB L1) with Philadelphia chromosome and rearrangements of bcr gene with deletion of 5' side and of TCR delta gene.

The expression of EGF receptor, c-neu oncogene product, and of transferrin receptor in 13 cases of chordomas (9 without metastasis and 4 with metastasis) was examined immunohistologically by the B-SA method using specific antibodies to these proteins. Tumor cells in 5 cases without metastasis and 3 cases with metastasis were positive for EGF receptor, those in 7 cases without metastasis and all cases with metastasis were positive for c-neu product, while those in all cases were negative for transferrin receptor. These results suggest a possible link between the expression of cellular genes associated with the growth of neuroectodermal cells and the growth of chordoma known to arise from the embryonal rest of the notochord.

We have utilized the polymerase chain reaction (PCR) to sensitively detect the minimal residual leukemia in 24 patients who were hematologically normal and were negative for Ph1 chromosome after bone marrow transplantation (BMT). None of the patient showed breakpoint cluster region (BCR) gene rearrangement by Southern blot analysis, however, PCR technique revealed the bcr/abl mRNA in 13 of 24 patients. In patients who received CY + TBI as a conditioning regimen, 6 out of 8 patients were detected bcr/abl mRNA by PCR. On the other hand, patients who received high dose AraC+ CY + TBI or busulfan + CY as a conditioning, bcr/abl mRNA was detected in 4 out of 9 patients and 1 out of 5 patients, respectively. There was no clear association between the presence or absence of graft versus host disease and the detection of bcr/abl mRNA by PCR.

Polymerase chain reaction (PCR) is a recently-developed technique capable of amplifying a specific nucleotide sequence and has been proved highly sensitive enough to analyse a small amount of DNA. This prompts us to study minimal residual disease (MRD) which is undetectable by conventional morphological or cytogenetic analysis. Using this technique, tumor specific chimeric DNA or mRNA were detected in patients with follicular lymphoma with t(14; 18) or in Ph1 positive ALL in complete remission, respectively. PCR was also useful to monitor MRD after treatment of bone marrow cells with monoclonal antibody. In sex-mismatched bone marrow transplantation, mixed chimerism was well documented by using repeated sequences which is unique to human Y chromosome. These results suggested that PCR offers a great help for detecting MRD or mixed-chimerism following bone marrow transplantation.

A multi-step model of the carcinogenesis for adult T-cell leukemia-lymphoma (ATL) has been reviewed here according to the authors observations. Epidemiological studies demonstrated that in most japanese ATL cases the disease developed from those who acquired infection of the causal agent, human T-cell lymphotropic virus type 1 (HTLV-1), during early infancy, probably through breast-feeding. Therefore, the latency period between the acquisition of HTLV-1 and clinical manifestation of ATL can be represented by the age of disease onset. It has been demonstrated by us that the age distribution of ATL onset can be described by a single Weibull distribution function, which is considered to be a feasible mathematical model for multistep carcinogenesis. Based on the present model, it is assumed that age-dependent accumulation of nearly five leukemogenic events, most likely somatic mutations, within the target cell(s) might be required prior to the disease onset.

The present study was undertaken to evaluate the nuclear DNA content of esophageal cancer cells consequent on the process of growth and progression of the cancer. In experimental animal studies, 47 esophageal cancers were induced in male Wistar rats by oral administration of N-amyl-N-methylnitrosamine (AMN) and were analysed in the study of ploidy patterns. The study was also carried out to determine the DNA content in the ploidy patterns in man. Primary tumors associated 421 metastatic lymph nodes in the 62 patients with thoracic esophageal cancer were subjected for the study of ploidy patterns. The nuclear DNA content was determined by means of flow cytometry. In the study of the experimentally-induced esophageal cancer in rats, aneuploidy was found in 18% at a depth of submucosa, 30% at proprial muscle, 59% at adventitia, and in 50% at a depth of neighboring structures, respectively. Clinically in man, the incidence of DNA diploidy and aneuploidy in the 62 primary cancers was 56% and 44%, respectively. In the 421 metastatic lymph nodes, diploid was found in 73% and aneuploid in 11%, while the combination of diploid and aneuploid was observed in 16%. Difference in the DNA index (DI) between the primary cancers and metastatic lymph nodes was found in 29 cases (46.8%), and the difference increased with progression of the cancer. Two hundred and ninety seven metastatic lymph nodes of 29 cases were subdivided into 4 groups based on the extent of the cancerous nests, and the DI value was found to be increased in proportion to the extension. With the results, the DI value of esophageal cancer appeared to be changed dependently by the variation of cell populations in the cancer or in the DNA content of the cancer cells.

Using the megakaryocytic leukemia cell lines, K-562 and CMK established from a Down's patient with acute megakaryoblastic leukemia, we studied the changes of antigen expression, cytosolic Ca2+ mobilization, thromboxane (TX) A2 formation and gene expression during megakaryocyte differentiation. We found that thrombospondin synthesis and platelet factor (PF)-4 gene expression were specific for mature megakaryoblasts, whereas collagen unresponsiveness and prostaglandin E1-induced Ca2+ mobilization were noted in immature megakaryoblasts alone. This experiment shows that functional and genetic analysis are useful for characterizing the leukemic megakaryoblastic cells. We analyzed the clinical, hematologic and genetic features of 4 patients with M7, and acute megakaryoblastic transformation of CML, MDS and essential thrombocythemia. In two patients, prednisolone and 6-MP were effective in cytoreduction. In 3 patients with increased platelet counts, normal CFU-Meg formation, the megakaryoblasts with platelet production, or the coexistence of immature megakaryoblasts with mature megakaryocytes were observed, thus indicating that some megakaryoblastic leukemia cells still have the capacity of differentiation. One patient had megakaryoblastic cells with PF-4 gene expression. These clinical findings suggest that the megakaryoblastic leukemia could not be characterized as usual leukemia and a more sensitive marker is required to differentiate leukemic megakaryoblasts from normal megakaryoblasts.

Correlated studies on the cellular kinetics in association with the nuclear ploidy pattern and the pathological morphology of advanced colorectal cancers have been carried out using DNA-RNA cytofluorometry with an AO stain. The results showed that advanced colorectal cancers could be separated into the 3 distinct groups that follow: group I, involving a diploid cell population; group II, a polyploid cell population; and, group II', an aneuploid cell population. The group I type was located in the entire colorectal region, where as groups II and II', were in the sigmoid and rectum. Three groups were mostly Borrmann type 3 cancers, and the gross pathological classification was not found to be related to the ploidy pattern.

One family of 17 cases of general fibrosis syndrome was reported. Four out of five patients examined with CT and/or MRI revealed cavum vergae or cyst of the cavum septi pellucidi or mega cisterna magna, which were characterized by existing on the midline of the brain. The patient with cavum vergae had also platybasia. In one patient, surgical correction of vertical deviation and blepharoptosis of both eyes was performed. There was abnormal insertion of the superior and inferior rectus muscles in posterior and nasal direction, and adhesion of the superior oblique muscle to the superior rectus muscle at the point of it's insertion was found in both eyes. Histopathological findings of the extraocular muscles of two patients showed mixture of relatively normal muscle tissues and vast amounts of collagen fibers. Electron microscopically many glycogen granules were found in muscle fibers. From these findings, this syndrome may be based on failure in development and differentiation.

Using three chromosome 18-specific DNAs, rearrangements of a bcl-2 gene were detected in 20 (44%) of 45 Japanese patients (pts) with follicular lymphoma (FL) and 3 (8%) of 36 pts with diffuse large cell lymphoma. The 21 pts had t (IgH; bcl-2), and of the remaining two who did not display it, one had chromosome t(14; 18). Compared with the findings in American pts, the lower frequency of t(14; 18)-positive lymphoma could reflect a difference in the incidence of overall nodal B lymphoma between Japan and the United States. 11 of 18 pts with t(14; 18)-positive FL expressed CD10 antigen on the cell surface and all 12 pts with t(14; 18)-negative FL did not, and the difference is statistically significant, indicating that t(14; 18)-positive Japanese FL is the same disease entity as most of American FL. Extra 18q- chromosome found in the advanced grade diseases of t(14; 18)-positive lymphoma results in amplification of the rearranged bcl-2 gene on 18q-, suggesting that the change is closely associated with transformation of FL carrying t(14; 18). It is thus possible of international realization to institute the prognostic factors and the treatment strategy for conquering t(14; 18)-positive lymphoma.

Although determination of chromosomal abnormalities may be of limited usefulness for the diagnosis of leukemia, the recent advances in the molecular mechanism associated with chromosome aberration has been rapid. Chromosome translocation in Burkitt lymphoma and chronic myeloid leukemia was the most striking evidence for the oncogene activation. Other specific chromosome abnormalities for FAB-classified leukemias are also known. Translocated type of chromosome abnormalities between immunoglobulin or T-cell receptor genes and oncogenes may also affect the T and B-cell leukemogenesis. However, the role of trisomies found in human and experimental leukemias and the gene dosages had been thought to be most important until 1982, has not been unclear. Many types of phenotypically heterogeneous leukemias have been reported. t(4 ; 11) acute leukemia is one such leukemia which shows early B-cell and myelomonocytic nature. Heterogeneous leukemias have been called biphenotypic, hybrid and acute mixed leukemias. The terminology must be used the unified. Recent trials to use paraffin-fixed tissues and bone marrow smear for molecular analysis has been successfully reported. Basic analysis on the DNA degradation mechanism revealed the enzymatic activity might play an important role before the complete fixation.

Von Recklinghausen's neurofibromatosis is a congenital anomaly due to maldevelopment of the neurectoderm and mesoderm. The disease is known to be commonly associated with other tumors of the neural system. We experienced a case in which two different types of spinal tumors (meningioma and schwannoma) appeared together with Von Recklinghausen's disease. The patient was a 54 year-old female. Her chief complaint was slowly progressive lumbar back pain of about a 10-year duration. She was admitted to our hospital after developing such clinical symptoms as paraparesis, bladder and rectal incontinence, and pain in the soles of her feet. The CT study demonstrated a massive tumor at the TH12-L2 levels accompanied with marked scalloping of the posterior vertebral body. The tumor removed was, histologically, meningioma. During surgery, another small bean-size tumor was incidentally found originating from the dorsal root, and it was identified by histological study as typical schwannoma. It has been reported that the incidence rate of spinal tumors in von Recklinghausen's disease is approximately 4%. Although a case has been reported in which different types of tumors developed in both the cranium and the spinal canal, our case as described above is considered extremely rare as far as we can find in reference literature. The pathogenic mechanism of vertebral scalloping as encountered in our case was thought to be based on interactions between hypoplasia of the supporting tissue and pressure generated by the tumor and CSF.

We report a case of nasal lymphoma with characteristics of natural killer (NH) cells. A 44-year-old man was admitted to our hospital because of right nasal obstruction. Physical examination revealed a tumor in the right nasal cavity and swelling of the right cervical lymph nodes. Histopathological examination showed diffuse medium sized cell lymphoma. The neoplastic cells expressed CD2, NKH-1, HLA-DR and leukocyte common antigen, but lacked other T-cell, B-cell and myeloid markers. They were in germline configuration for immunoglobulin heavy chain and T-cell receptor genes by southern blot analysis. These findings suggest that they are derived from NK cell.

Translocations involving band q32 on chromosome 14 are commonly identified as a 14q + marker-chromosome and the involvement is the single most frequent abnormality in various types of lymphoid cancer. Based on these findings, we have proposed that the 14q + marker-positive lymphoid cancer should be divided into subclasses according to the precise translocations, each of which is a primary chromosome change, and the subclass initiates serial processing of the karyotype evolution associated with the development of more aggressive lymphoid cancer. The proposal is strongly supported by recent advances in molecular genetics. Genes or unknown DNA sequences, which had evolutionally been conserved, were isolated from the breakpoint on the partner chromosome of the 14q+ translocation, and the DNA sequences identified to juxtapose to the IgH gene locus on chromosome 14 band q32 accompanied by individual translocation. Thus, we can subclassify the lymphoid cancers marked with chromosome changes involving the loci of other functional genes.

Adult T-cell leukemia-lymphoma (ATL) is a unique T-cell leukemia-lymphoma that is closely associated with human T-cell leukemia virus type I (HTLV-I). HTLV-I is also associated with a benign disease, HTLV-I associated myelopathy. Main routes of the infection is through breast milk feeding from carrier mother to her baby, from carrier husband to wife, and by blood transfusion of carrier blood to recipient patients. The presence of anti-HTLV-I antibody is a definite sign of the carrier of HTLV-I, then screening of the antibody has been introduced with success to prevent the HTLV-I infection. Mechanism of leukemogenesis by the virus is not known. It is important to note that age-dependent occurrence of HTLV-I associated ATL can be simply described by a Weibull model. This suggests that ATL leukemogenesis might be the result of accumulation of numerous critical events, most likely somatic mutations, which are estimated to be five from the analysis. Although HTLV-I infection plays a primary role in the pathogenesis of ATL as an initiator, it may be only a prerequisite for accumulation of later events. The presence of HTLV-I negative ATL suggests that factor(s) other than HTLV-I infection may be involved in the leukemogenic process of ATL.

Using DAPI-DNA cytofluorometry, the author analyzed nuclear DNA content of formalin fixed, paraffin embedded, glioma material obtained from 14 glioma cases at surgery. Sections of 10 microns were deparaffinized. Following simultaneous DAPI (4,6-diamidino-2-phenylindole dihydroporphyrin chloride)/HP (hematoporphyrin) staining, DAPI binds DNA and DNA-DAPI complexes emit blue fluorescence when exited by ultraviolet (UV) light. Through Zeiss fluorescence microscope, the author measured nuclear fluorescence intensity with histological verification of glioma cells. A DNA histogram was obtained with fluorescence intensity recorded on the abscissa and number of cells plotted on the ordinate. Samples of 20 normal non-neoplastic astrocytes taken from apparently normal brain tissue included in the histological slide were used as diploid (2 C) control. Based on DNA content, tumor cells were classified into 4 groups: N-group composed of cells with 2 C DNA content (normoploid), S-group with less than 2 C (hypoploid), L-group more than 4 C (hypertetraploid), I-group between 2 C and 4 C (intermediate ploidy). Intermediate ploidy was significantly higher and normoploid was significantly lower in glioblastoma compared with those of benign astrocytoma. Thus, DNA content and histological malignancy were well correlated. Due to limitation of measuring diaphragm of turret in the microscope, some extra large cell could not be included in it and was excluded from the measurement.(ABSTRACT TRUNCATED AT 250 WORDS)

The relationship between nuclear DNA content determined by cytofluorometry in the primary focus of breast cancer and the survival rate was analyzed to clarify the prognostic significance of nuclear DNA content in breast cancer. The relationships of the ploidy pattern and the frequency of polyploid cells (4c or above) with the survival rate were studied in patients who underwent extended radical mastectomy and were comparable in the clinical stage and other prognostic factors. The survival rate was significantly lower in those of the non-diploid type showing no prominent peak at 2c or those in whom the frequency of polyploid cells was more than 30% than in those of the diploid type with a prominent peak at 2c or those showing few polyploid cells, even when the disease stage (Stage II by TNM classification and stage III by Tnm classification), histological lymph node metastasis (n (+), n1 beta), and histological type (papillotubular carcinoma, scirrhous carcinoma) were identical. From these findings, nuclear DNA content is considered to be a parameter of the malignancy of breast cancer and to have clinical significance as an important prognostic factor.