Molecular detection of tumor cells at diagnosis invading the bone marrow and peripheral blood of patients with aggressive or indolent lymphomas.

We studied tumor cell invasions of bone marrow and peripheral blood in patients with various types of advanced non-Hodgkin's lymphoma by amplifying complementarity determining region III using the polymerase chain reaction (PCR) method and developing patient-specific probes. After molecular engineering, we could detect tumor cells in bone marrow from seven of 11 cases and in peripheral blood from six of 11 cases, despite negative results in four cases studied morphologically. Indolent cases were more likely to yield positive results than aggressive cases. The reason may be different biological behaviors among the histological types.

[Marked thrombocytopenia after high-dose intravenous gamma globulin in a pregnant woman with idiopathic thrombocytopenic purpura].

A 35-year-old pregnant woman had thrombocytopenia with a platelet count of 6.3 x 10(4)/microliter. After her third normal delivery, peripheral blood studies revealed that the patient had a normal Hb concentration and leukocyte count, with mild thrombocytopenia. A diagnosis of idiopathic thrombocytopenic purpura (ITP) was made based on the high megakaryocyte count of 338/microliter and PAIgG of 40.8 ng/10(7) cells in January 1995. The patient was followed without treatment. She was 9 weeks pregnant on June 7, 1996, and desired an abortion. Her platelet count was 6.3 x 10(4)/microliter, leukocyte count 8,600/microliter, and Hb 13.7 g/dl at the time. She was given high-dose intravenous gammaglobulin (Globenin-I) at 400 mg/kg/day for 5 consecutive days. The platelet count was found to have decreased markedly, to 0.9 x 10(4)/microliter on June 11. The percentage reduction in the Hb concentration, leukocyte count, and platelet count after gammaglobulin treatment was 11.7%, 46.6%, and 85.8%, respectively. The PAIgG titer had increased to 181.2 ng/10(7) cells on June 17, but hypergammaglobulinemia was suspected. The patient was started on prednisolone on June 24, and an abortion was performed on July 29. The mechanism of thrombocytopenia after infusion of Globenin-I was unknown. We suspect that Globenin-I treated with polyethylene glycol was one of the possible causes of myelosuppression in this case.

[Myelodysplastic syndrome accompanied by i(17) (q10) anomaly following pure red cell aplasia and transient myeloproliferative stage].

A 71-year-old man was given a diagnosis of pure red cell aplasia (PRCA) in May 1995. However, immunosuppressive agents, including prednisolone, azathioprine, and cyclosporin A, were not effective, and he required frequent red cell transfusions. In September 1995, leukocytosis and thrombocytosis developed (peaking at 10,100/microliter white cells and 98.1 x 10(4)/microliter platelets, respectively, in November 1996). Conversely, the patient's peripheral blood count began to decrease in July 1996, and pancytopenia progressed thereafter i(17) (q10) chromosomal abnormality of bone marrow cells was detected in November 1996. The patient was readmitted due to the progression of thrombocytopenia (1.2 x 10(4)/microliter). His bone marrow has 16.6% blasts, and a diagnosis of myelodysplastic syndrome (MDS) was made. The patient died in November 1997. His hematological state demonstrated significant changes in a relatively short period and severe hypoerythropoiesis and eosinophilia of the bone marrow persisted throughout the clinical course. These findings suggested that a common deranged stem cell was the origin of 3 different states; PRCA, chromic myeloproliferative disorder, and MDS. The i(17) (q10) anomaly may have caused the acute proliferation of blasts and pancytopenia.

A staging system for multiple myeloma based on the morphology of myeloma cells.

The morphology of myeloma cells is reported to be one of the prognostic factors in multiple myeloma (MM) patients. We analyzed the prognostic factors, including morphological classification, in 292 patients with MM in order to select poor-risk patients who should be considered candidates for early intensive chemotherapy, including stem cell transplantation. Multivariate analysis was applied to 90 patients diagnosed between 1989 and 1996, because serum beta-2-microglobulin (beta2M) has been measured regularly since 1989, and showed that serum albumin, serum beta2M, and the morphology of myeloma cells predicted survival. According to these factors, patients were divided into 3 risk groups; a high-risk group (14%), a intermediate-risk group (46%) and a low-risk group (40%). There were significant differences between survival times in these 3 groups (median survival: high-risk, 16; intermediate-risk, 22; and low-risk, 44 months).

Differences of IgG and IgM antibodies to phospholipids and APTT among non-pregnant and pregnant women.

OBJECTIVE: The objective of our study was to assess the difference in IgG and IgM antibodies to cardiolipin(CL), phosphatidylserine(PS), and APTT among non-pregnant and pregnant women. METHODS: IgG and IgM antibodies to CL and PS among 102 healthy non-pregnant women and 154 healthy pregnant women were measured by an ELISA. The activated partial thromboplastin time(APTT) was measured in 67 healthy non-pregnant women, in 67 healthy women at 10-14 weeks of gestation, and in 67 healthy women at 30-32 weeks of gestation. We compared the titer of +2SD in each group. RESULTS: The titers +2SD of IgG antibodies to CL and PS in pregnant women were lower than in non-pregnant women (p < 0.05), whereas the values of the IgM antibody to CL and PS were the same in pregnant and non-pregnant women. The APTT in pregnant women was significantly shorter than in non-pregnant women (p = 0.0001). CONCLUSION: Standard criteria for the positivity of the IgG antibody to CL and PS and the prolongation of APTT in pregnancy should employ the values of normal pregnant women rather than those of non-pregnant women.

[Idiopathic CD4+ T-lymphocytopenia terminating in Burkitt's lymphoma].

A 33-year-old male was admitted because of severe neutropenia (55/microliter) in 1985. A diagnosis of autoimmune neutropenia was not made. CD4+ T-lymphocyte count was 128/microliter and CD4/CD8, 0.1, though the titers of immunoglobulins were in normal range. Nine times examinations of CD4+ T-lymphocyte count resulted in 5 times lower counts than 300/microliter between 1985 and 1993. His HIV (type 1 and type 2) studies were negative. He suffered from EB virus infection in 1987. The titers of VCA-IgG and EA-IgG had been elevated for half a year after that. He was operated polyp-like tumors in bilateral nasal cavities on February in 1994. The diagnosis was Burkitt's lymphoma that was also demonstrated by in situ hybridization using EB virus RNA. He was treated with a modification of COPBLAM III, but he died on June in 1994. Autopsy revealed tumor cell infiltrations in multiple organs. We suspect that he developed Burkitt's lymphoma 82 months later, when he had complicated EB virus infection in condition of idiopathic CD4+ T-lymphocytopenia caused by unknown origin.

Hepatitis B virus carriers in the treatment of malignant lymphoma: an epidemiological study in Japan.

BACKGROUND: Hepatitis B after the withdrawal of cytotoxic chemotherapy in hepatitis B virus (HBV) carriers is well known and may lead to fatal hepatic failure. We retrospectively analyzed the prevalence of HBV carriers, the incidence, and the risk factors of hepatitis B in the treatment of malignant lymphoma. PATIENTS AND METHODS: HBV carriers were defined as patients with positive HBs-antigen, either with normal or abnormal serum aminotransferase level at patient presentation. Questionnaires to the members of the Japan Lymphoma Treatment Study Group included general information, details about HBV carriers, and further information about hepatitis B. RESULTS: Among 1380 patients collected from eight institutions, 45 patients (3.26%) were determined to be HBV carriers, Hepatitis B developed in 17 of the HBV carrying patients (37.8%). Seven of those 17 (41.2%) died of hepatic failure. Hepatitis developed at a high rate in patients who were negative for HBe-antigen (50%), and who had received second- or third-generation chemotherapy (63.2%). CONCLUSION: We confirmed that hepatitis B developed with high frequency in HBV carriers with malignant lymphoma. Moreover, hepatitis often resulted in fatal hepatic failure. It is necessary to prevent the hepatitis B developing in HBV carriers when receiving intensive chemotherapy for malignant lymphoma.

[A HBV carrier with fulminant hepatitis complicated by ATL, multiple myeloma and thyroid cancer].

A 75-year-old female, born in Tochigi Prefecture, was admitted because of lumbago in August of 1991. The leukocyte count was 11,800/microliters with 22.5% atypical lymphocytes. We demonstrated a lymphocyte surface marker, ATL-associated antigen, and proviral DNA. We also identified 2.60 g/dl of serum monoclonal protein, found to be IgG, lambda type, and punched out lesions in the skull. We made a diagnosis of ATL. She was also a HBV carrier. The patient was treated with a modification of CHOP therapy, because of increasing atypical lymphocytes in the peripheral blood in November of 1992. She died of acute hepatitis, suddenly, in March of 1993. Autopsy revealed multiple myeloma, fulminant hepatitis and occult thyroid cancer in addition to ATL.

Ten-year survivors with multiple myeloma.

Of 130 Japanese patients with symptomatic multiple myeloma who were treated between 1970 and 1989, nine (6.9%) patients survived for more than 10 years. Younger age, low and intermediate tumour mass, chemotherapy with cyclophosphamide, the disappearance of myeloma protein, and a positive response to retreatment were correlated with long-term survival.

Altered expression of blood group A and H antigens on red cells from an acute leukemic patient.

Alternate expressions of the blood group A and H antigens on red cells are described in a patient with acute myelocytic leukemia. The patient's red cells showed mixed field agglutination with anti-A and anti-H sera and lectins, and no agglutination with anti-B serum. The agglutinability of the A red cells with Dolichos biflorus lectin was between A1 and A2 (A intermediate). Inagglutinable red cells were separated with anti-A agglutinin, and the proportion was about 80% of total cells. The agglutinating activity with Ulex europaeus anti-H of red cells, which were inagglutinable with anti-A, was 16 times weaker than that of group O cells. The weaker reaction with Ricinus communis lectin and the stronger reaction with Psathyrella velutina lectin on the inagglutinable cells with anti-A than those on the group O cells suggest that fucosyl alpha (1-2) and galactosyl beta (1-4) residues at the nonreducing end of carbohydrate chains of H antigens on the red cells were diminished, and N-acetylglucosaminyl beta (1-3) residues were sequentially exposed. His saliva contained A and H substances in normal amounts of a secretor. Serum alpha-N-acetylgalactosaminyltransferase activity which converts O red cells to A red cells was the same as those in sera from A1 individuals. These results suggest that the synthesis of H precursors is partially blocked in this patient's red cells.

Relatively low prevalence of human papillomavirus 16, 18 and 33 DNA in the normal cervices of Japanese women shown by polymerase chain reaction.

Ninety-two cervical scrapes and tissues, obtained from cytologically or histologically normal cervices of Japanese women, were examined for the presence of human papillomavirus (HPV) 16, 18 and 33 DNA by the polymerase chain reaction (PCR) method. Five out of 92 cases were HPV 16 DNA-positive, but neither HPV 18 nor 33 DNA was detected. The HPV (type 16, 18 and 33) prevalence rate in pregnant women, including postpartum, was 10% (3/31), which was higher than that in non-pregnant women. In two HPV 16-positive cases, we detected HPV 16 DNA again 2 months later. HPV (type 16, 18 and 33) prevalence in normal cervices was shown to be relatively low. However, it is very important to follow up the HPV-positive cases in cytologically normal cervices in order to elucidate the relation between HPV infection and the progression of cervical cancer.

Human papillomavirus type 31 DNA detected in part of the dysplasia but in no part of the squamous metaplasia in a specimen taken from one patient.

Using in situ hybridization, human papillomavirus (HPV 6, 16, 18, 31, 33) DNAs were detected in a cervical severe dysplasia accompanied by squamous metaplasia. It was found that, only HPV 31 DNA was harbored in the cervical severe dysplasia, but HPV DNAs were not identified in a lesion of squamous metaplasia. The in situ hybridization method will be of use, therefore, when dysplasia with squamous metaplasia or other lesions are examined for HPV DNA. In a cervical smear, HPV 31 DNA could be detected on the nuclei of dysplastic cells, so this method is applicable to cervical smears. If squamous metaplasia is to be considered as a precursor lesion to cervical dysplasia, the HPV DNA harbored in the dysplasia must also be detected in the accompanying squamous metaplasia. Our results suggested that not all squamous metaplasias were involved with HPV, as far as we were able to detect using five types of HPV DNA probe.

[A case report of Buschke-Lowenstein tumor (giant condyloma)].

A 28-year-old woman with diabetes mellitus and alcoholic hepatitis presented a rare case of Buschke Lowenstein tumor, or giant condyloma of vulva. HPV DNA of this tumor was detected by in situ hybridization using tritium labeled HPV 6 b, 16 and 18 DNA. This tumor appeared to harbour HPV 6 b DNA, but other HPV DNAs were negative. The distribution of HPV 6 b DNA was detected in the nucleus of the squamous epithelium showing koilocytosis. Total vulvectomy and resection of the clitoris were done. The postoperative course was uneventful and there has been no recurrence of tumor so far.

The detection of human papillomavirus (HPV) in a case of minimal deviation adenocarcinoma of the uterine cervix (adenoma malignum) using in situ hybridization.

HPV-16 and HPV-18 DNA sequences were detected in a case of minimal deviation adenocarcinoma (MDA), a rare pathological type of uterine cervical cancer. This result suggests HPV to be associated with MDA as well as with other pathological types of cervical cancer.

[The sensitivity of in situ hybridization for detection of human papillomavirus].

We studied a sensitivity of HPV DNA detection by in situ hybridization method using 3H labeled HPV DNA. The materials were CaSki cells and SiHa cells which were derived from as a negative control. The total cellular DNAs extracted from these cell lines were estimated copy numbers of HPV 16 DNA using Southern blot hybridization. In our result, CaSki cell has 400 copies/cell, SiHa cell were appeared to have 1-5 copies/cell. Simultaneously these cells were fixed by periodate-buffered lysine-paraformaldehyde-glutaraldehyde (PLPG) and were detected HPV 16 DNA using in situ hybridization. We detected HPV 16 DNA in CaSki cells and SiHa cells by in situ hybridization also. We concluded that the sensitivity of our in situ hybridization technique is 1-5 copies/cell.

The physical state of human papillomavirus 16 DNA in cervical carcinoma and cervical intraepithelial neoplasia.

Cervical carcinomas and cervical intraepithelial neoplasias (CIN) were analyzed for the presence of human papillomavirus (HPV) DNA using Southern blot hybridization. Of the five HPV types examined (HPV types 6, 11, 16, 18, and 33), HPV 16 DNA was detected most frequently. In most HPV 16-positive carcinomas examined, HPV 16 DNA was present in an integrated state in cellular DNA with or without the coexistence of episomal species. In one case, however, only episomal species were detected. Among seven cases of HPV 16-positive CIN, four contained HPV 16 DNA only in the episomal state and the rest contained HPV 16 DNA only in the integrated state, but the coexistence of both states was not found. These results suggest that the integration of HPV 16 DNA is not necessary for cells to become malignant, although it is frequently associated with malignant cells.

[The presence of HPV DNA in squamous metaplasia, dysplasia and carcinoma in situ of the uterine cervix].

HPV DNA was detected in 11% (4/36) of cervical squamous metaplasia, 43% (6/14) of mild dysplasia, 39% (7/18) of moderate dysplasia, 50% (7/14) of severe dysplasia and 50% (7/14) of CIS. The incidence of HPV16 DNA increased with the severity of dysplasia through CIS. It was distributed almost evenly in the nuclei of the total epithelial layers in severe dysplasia and CIS. Especially in CIS, HPV DNA was found significantly in the nuclei of both basal and parabasal cells, suggesting the possibility of involvement in the carcinogenesis of cervical cancer. The presence of HPV DNA in squamous metaplasia (36 cases) was investigated in three different pathological situations. Group 1 was squamous metaplasia in squamo-columnar junction; Group 2 in the polyp; and Group 3 accompanied by dysplasia or CIS. HPV DNA was positive only in squamous metaplasias with polyps. In these HPV DNA positive cases, HPV DNA was distributed not only in the metaplastic lesion, but also in the adjacent stromal lesion.

[Four cases of an extramedullary plasmacytoma of the head and neck].

From 1987 to 1988, four patients with solitary, extramedullary plasmacytomas of the head and neck were seen at the Gunma Cancer Center Hospital. With reference to such plasmacytomas, we have reviewed the medical literature regarding these tumors and mainly present the results of radiation therapy with respect to these patients. One patient, who had a local recurrence after surgical treatment, underwent radiation therapy (36Gy in four weeks) that resulted in good local control of the disease. Two patients developed multiple painful bone lesions during their long follow-up periods without manifesting any abnormal laboratory findings, suggesting multiple myeloma. It was found that radiation therapy (less than 30 Gy) was useful for the palliation of painful lesions.

Thyrotropin measured by the immunoradiometric assay in low birth weight infants.

In 37 infants, the blood levels of TSH were determined by the immunoradiometric assay (IRMA) and the relation between TSH and thyroid hormone was evaluated. The ranges of gestational age (weeks) and birth weight (g) of infants were 28-42 and 982-3,650, respectively. The birth weights of 19 infants were below 2,500 g. The free T4 levels in the low birth weight (LBW) infants were lower than those of the normal infants and significantly correlated to the birth weight (r = 0.64, P less than 0.01) and gestational age (r = 0.58, P less than 0.01). In addition, free T4 levels were significantly correlated to the levels of total T4 (r = 0.66, P less than 0.01). The concentrations of TSH measured by IRMA method were significantly correlated to those of free T4 (r = 0.51, P less than 0.01). From these data, we consider that the transient hypothyroxinemia observed frequently in LBW infants might be a physiological reaction regulated by hypothalamus and that thyroid hormone treatment should be avoided.