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A cellular association demarcated by two perpendiculars which were drawn between adjacent bundles of elongate spermatids from the tubular lumen to the basement membrane, was made the unit of histometrical observation in this study (provisionally called a "column"). Cell counting revealed that the average numbers per column of various types of germ cells do not show any significant differences among 5 fowls and between paired testes. The frequency of spermiogenic steps (numbered 1-8) was investigated in each column. A definite and common pattern was found in the frequency distribution in the 5 fowls observed. A relationship between spermiation and younger spermatid steps was also investigated in each column. The spermiation was found at different steps, but most frequently at step 2 (30.6%). Based on these observations and referring to other author's information, an average time interval between two successive spermiations was calculated roughly at 3.3 +/- 1.2 days. Theoretically, this value is equal to an average length of one epithelial cycle. Such a variable cycle may have caused irregular cellular associations in this species.

It was investigated in the domestic fowl, whether a mechanism for a synchronous start of the proliferation of the differentiating spermatogonia in a transversely cut seminiferous tubule (called the "co-ordination" between germ cell association) operates effectively in this species. A trapezoid-shaped cellular association demarcated by two perpendiculars from the tubular lumen to the basement membrane was the unit of observation (provisionally called a "column" Each column is composed of a bundle of elongate spermatids and the neighbouring, various types of germ cells). The extent of closeness of the co-ordination was examined in a total of 2,650 columns, based on the continuity of the spermiogenic step-number of the youngest generation of spermatids within adjacent columns. In no case did all columns in a tubular cross section show the same-step spermatids. In most cases (50.2% of 2,650 columns), the same number was kept in only 2 to 6 adjacent columns. The arrangement of different but consecutive numbers (e.g., 1-2-3 or 1-2-1) was observed in a total of 882 columns. The co-ordination mechanism, therefore, is considered to work, albeit incomplete, among a limited number of adjacent columns in this species.

A determination method for the penicillin antibiotic aspoxicillin in broncho-alveolar lavage fluid has been developed, involving high-performance liquid chromatography and post-column photolysis. The method enabled the determination of aspoxicillin at concentrations below 100 ng/ml, with a linear calibration curve at concentrations of 1-1000 ng/ml for 100 microliters of lavage fluid. The detection limit was 1 ng/ml at a signal-to-noise ratio of 3:1.

The activity of pulmonary lymphocytes was evaluated by the detection of interleukin-2 (IL-2) receptor alpha mRNA expression in lung cancer patients associated with diffuse interstitial shadow on roentgenograms of their lungs. Reverse transcription coupled with the polymerase chain reaction was used to detect mRNA expression. In 5 of 6 patients, IL-2R alpha mRNA expression was increased in pulmonary lymphocytes compared with 4 normal controls. The expression in this mRNA in peripheral blood lymphocytes was almost undetectable in either normal controls or these patients. These results suggest that pulmonary lymphocytes in patients with lung cancer associated with diffuse interstitial shadows are activated and may promote the inflammatory process generating pulmonary fibrosis.

Three leukemia cell lines, BALM-6, -7 and -8 were established from the bone marrow of a patient with acute lymphoblastic leukemia. Based on immunophenotypic and the cytogenetic analysis and on the expression of immunoglobulins on both the cell surface and in the cytoplasm, BALM-6, BALM-7 and BALM-8 are clonal leukemic cell populations of B cell origin.

Serum progesterone (P) concentrations using P-EIA kit (Ovucheck, Cambridge Life Science Co., Ltd.) were examined in 8 captive and 7 free-ranging female Hokkaido brown bears (Ursus arctos yesoensis). The intra- and inter-assay coefficients of variation were 8.9%, 12.6% and 16.6%, 22.7%, respectively, based on 2 serum samples. There was a significant correlation between EIA and radioimmunoassay results based on 64 serum samples (r = 0.725; p less than 0.01). Serum P concentrations were examined in 5 pregnant, 2 solitary non-pregnant bears and a lactating non-pregnant bear in captivity. Annual changes of P levels in pregnant bears were observed as a small elevation during the mating season (May-June), a re-elevation in September-October and a sharp elevation in November-December. The sharp elevation was suspected to reflect changes when implantation occurred. Annual changes of P levels in solitary non-pregnant bears were similar to those in pregnant bears. An annual change of P levels in a lactating non-pregnant bear maintained levels under 5 ng/ml. Two of 7 free-ranging bears exhibited P levels over 1 ng/ml and the birth of cubs was confirmed in the following year in 1 of the 2 bears. P concentrations of other free-ranging bears exhibited less than 1 ng/ml, and these bears were considered to be non-pregnant. It was concluded that P-EIA kit was available for measuring P concentrations in Hokkaido brown bears.

Based on the immunophenotypic, cytogenetic and genotypic findings, two unique leukemia cell lines, NALM-24 and NALM-25, and an EBV-transformed "normal" B-lymphoblastoid cell line (B262) from a patient with ALL were established and characterized. NALM-24 and NALM-25 are unique in that expression of both show B cell and myeloid cell features with the t(9;22) chromosome in single clonal leukemic cell populations.

Based on the immunophenotypic and genotypic findings, this acute leukemia cell line, designated NALM-19, is unique in that a partial expression of both B-cell and myeloid cell features are present in this single clonal leukemic cell population. It is noteworthy that two "normal" EB virus-transformed B cell lines, B239 and B240, (paired with NALM-19) were established from the same leukemic blood.

Based on the morphological resemblance to megakaryocytes and upon the expression f the platelet specific glycoprotein antigen, CD61, it is highly l y that MOLM-1 is a clonal leukemic cell population of megakaryoblastic origin .

The prophylactic effect of the oral administration of high-dose amphotericin B syrup for the systemic fungal infection was studied in 36 patients with hematological neoplasms. Twenty nine patients received 2,400 mg/day of Amphotericin B syrup for during the remission induction therapy. One patient received 1,200 mg/day, 3 received 800 mg/day and 3 received 400 mg/day of Amphotericin B syrup. The prophylactic effect was recognized in 24 of 36 patients, 66.7%. As adverse effects gastrointestinal symptoms such as nausea and vomiting, hypochloremia and hypopotassemia associated with hypochloremia was observed in one patient, respectively, however, they were all controllable. The blood levels of Amphotericin B in patients received 2,400 mg/day was 0.092 +/- 0.055 micrograms/ml (n = 40) on 7th day and 0.110 +/- 0.046 micrograms/ml (n = 21) on 28th day, respectively. The administration of high-dose of Amphotericin B syrup is expected not only for the prophylaxis but also for the treatment of the systemic fungal infection.

Methotrexate (MTX), a folate antagonist and anticancer agent, was linked covalently to murine monoclonal anti-HLA IgG1 antibody (H-1) with the use of dextran T-40 as a multivalent carrier. As determined spectrophotometrically, the conjugate was composed of 9.20 mol MTX per 1 mol antibody. Of the substituted MTX, 46.3% (4.30 mol MTX per 1 mol antibody) exhibited inhibitory activity on dihydrofolate reductase. The MTX conjugated to H-1 [MTX-(H-l)] showed significantly stronger cytotoxicity against HLA-bearing cells than cells lacking HLA (p less than 0.001) in cultivation after a 2-hour exposure to the drugs. In the same experiment, free MTX showed equivalent cytotoxicity to cells both bearing and lacking HLA. When cells were cultivated for 3 days with the drugs, MTX-(H-l) also showed significantly stronger cytotoxicity than free MTX or MTX-control IgG conjugate against HLA-bearing cells (p less than 0.001), and the MTX-(H-l), MTX-control IgG, and free MTX showed equivalent cytotoxicity to cells lacking HLA. Our results indicate that the MTX-(H-l) conjugate binds to the cell surface antigen by its antibody action and exerts greater MTX cytotoxicity than free MTX in vitro.

Peripheral leucocytes from 10 patients with chronic myelogenous leukaemia (CML) were cultured in liquid medium and observed morphologically for 6 to 10 weeks. All cultures showed in increased proportion of mature basophilis at about 3 weeks and most cultures consisted predominantly of mature basophilis at about 5 weeks. The results indicate preferential basophilic differentiation of CML leucocytes in vitro and are consistent with basophilia frequently observed in CML patients.

One-way mixed lymphocyte reactions between lymphocytes from normal human donors and mitomycin C-treated cells from human leukemia T-cell, null cell, and B-cell lines were investigated. An Epstein-Barr virus (EBV)-negative Japanese Burkitt's lymphoma line and two EBV-positive normal lymphoid cell lines were studied in parallei. Normal lymphocytes were stimulated significantly by the cultured null cells and B-cells, but only slightly by the cultured T-cells. The stimulatory capacity of these two leukemia cell lines was approximately equal to that of the lymphoma and normal B-cell lines. The results suggest that not only leukemia B-cells but also leukemia null cells have stimulatory determinants in mixed lymphocyte culture.

The in vivo antitumor activity of NCS-immune immunoglobulin G (IgG) [Neocarzinostatin (NCS) conjugated with rabbit IgG antibody against a human leukemia cell line (NALL-1)] was evaluated in immunosuppressed newborn Syrian hamsters into which a transplantable human leukemia cell line, BALL-1 was implanted. After intraperitoneal (i.p.) injection of the conjugate, hamsters preinoculated i.p. with BALL-1 cells survived longer than hamsters treated with control solutions (p less than 0.01). The control solutions were NCS, immune IgG, a mixture of NCS and immune IgG, NCS-normal IgG conjugate and physiological saline. There was no change in body weight in the NCS-immune IgG-treated hamsters. The growth of subcutaneously (s.c.) implanted BALL-1 tumors was also inhibited by i.p. administration of NCS-immune IgG; however, the degree of inhibition was not significantly different from that obtained by administration of NCS alone, a mixture of NCS and immune IgG or NCS-normal IgG conjugate. These results indicate that NCS-immune IgG was effective against i.p. BALL-1 tumors, but was less effective against s.c. implanted tumors.

A 30 year old female patient diagnosed as acute lymphoblastic leukemia (ALL) with hand mirror like configuration of lymphoblastic-lymphocytic cells is reported. Although the leukemia was resistant to conventional chemotherapeutic regimens, the patient always looked well and survived for more than 20 months. Surface marker analysis showed that the cell was non-T, non-B, and not reactive to antiserum against common ALL antigen. A cytogenetic study of all the analyzable metaphases of the direct bone marrow preparation had a normal female karyotype. The clinical and hematological course is described. The immunological significance and the influence of hand mirror cell on chemosensitivity and prognosis are discussed.

Permanent human hematopoietic cell lines representing T-cell, B-cell and non T/non B (null-cell) leukemia have been established. Comparative analyses were made for their phenotype characteristics. A number of characteristics common within the 7 T-cell lines studied or distinct from other leukemia-type lines were described. Usefulness, validity and limitation of these findings are discussed in connection to the attempt at classification of ALL, CLL and blastic phase of CML. The great majority of CLL were SmIg+-B-cell leukemia and a single case of T-cell CLL was documented. Except 10% as T-cell ALL and a single case of B-cell ALL, the majority of ALL were found to be the non T/non B ALL. Nevertheless, little evidence was suggested from the present study in favor for a notion that the T-cell ALL and the non T/non B ALL are two distinct diseases.

A permanent hematopoietic cell line, designated NALM-1, was established from the peripheral blood of a patient who was in blastic crisis of Ph1-positive chronic myleocytic leukemia. By means of a panel of specific xenoantisera, the NALM-1 cells were found to express a specific antigen of acute lymphoblastic leukemia and blast leukemia-associated antigen. The cells exhibited no cell-surface receptors for sheep erythrocytes, IgG, or complement; neither cell-surface immunoglobulins nor cytoplasmic immunoglobulin were observed. Furthermore, normal T-cell or B-cell antigens, detectable by the antisera used in this study, were not found in the NALM-1 line.

Chromosome analysis was performed on cells from a patient of null cell lymphoma, well-differentiated type. A 14q12 translocation was observed in all the banded cells. In addition, there were multiple chromosome abnormalities. This case will be useful in considering the significance of the 14q1(1-3) translocation in malignant lymphoma disease.

A hairy cell leukemia (HCL) line, ZK-H, was established from peripheral blood of a 69-year-old male patient. The ZK-H cells and the patient's original hairy cells shared the same surface properties; both possessed membrane-bound IgG with kappa light chains and villous surface structures. The ZK-H line carried Epstein-Barr virus (EBV)-determined nuclear antigen, but the patient's fresh leukemic cells lacked this antigen. Morphologically, the ZK-H cells appeared lymphoblastoid and more primitive than the preculture cells. The ZK-H line had a hyperdiploid chromosome constitution of 47 and trisomy no. 2. The presence of membrane-bound immunoglobulin and of B-cell tropic EBV in this cell line provides further evidence for the B-cell nature of HCL in this patient

A 36-year-old man with adult T-cell leukemia gradually improved to the point of complete remission without any anticancer treatment. The patient was referred to our hospital because of the appearance of morphologically abnormal lymphocytes. Thirty percent of the peripheral blood white cells were morphologically abnormal lymphocytes which had lobulated or indented nuclei, and were E rosette-forming and mature T-cell antigen-positive. No hepatosplenomegaly, lymphadenopathy or mediastinal mass was noted. He had a cough with sputum one and a half years before entry. After admission, skin nodules occurred and were diagnosed histologically as non-Hodgkin's lymphoma, diffuse pleomorphic. Serum antibodies against an adult T-cell leukemia-associated antigen were detected at a titer of 1:40. Chest X-ray films, a pulmonary function test, bronchographic examination, bronchoalveolar lavage and transbronchial lung biopsy demonstrated no relationship between the pulmonary symptoms and the adult T-cell leukemia. After treatment with antibiotics and a beclomethasone inhaler, the symptoms decreased, and then the abnormal lymphocytes and skin nodules disappeared. The spontaneous, complete remission has continued for four years.

This study describes the establishment of two permanent leukemic B-cell lines (BALM-1 and -2) originating from the blood cells of a patient with a B-cell type of acute lymphoblastic leukemia. The cells of BALM-1 and -2 exhibited cell surface markers compatible with B-cell origin. The identity between the leukemic cells in vivo and those of the BALM lines was established on the basis of the karyotypic picture, including marker chromosomes 14q+ and t(12;17;?) and cell surface immunoglobulins (K, delta, and mu chain determinants).

Macromomycin (MCR), an unique membrane-reactive anticancer antibiotic, was incubated with murine monoclonal anti-HLA IgG1 antibody (H-1) in the presence of carbodiimide. The resulting mixture was fractionated with a Sephadex G-200 column. The first and second fractions were shown to contain MCR-(H-1) conjugate by the elution profile, as well as by the Sarcina lutea growth inhibition assay and Ouchterlony double-diffusion method. A membrane immunofluorescence test with anti-MCR and anti-mouse IgG antibodies demonstrated specific localization of MCR-(H-1) on the surface of HLA-bearing NALL -1 cells. MCR-(H-1) inhibited the growth of HLA-lacking NS-1 cells statistically less effectively than MCR alone (p less than 0.01). On the other hand, the conjugate and free MCR equally inhibited the growth and 3H-TdR incorporation of HLA-bearing NALL -1 cells. These results indicate that the antibody-bound MCR retained both MCR and antibody activities, and thus exerted antibody-targeting MCR cytotoxicity in vitro.

Fatty acids of plasma and erythrocyte membrane sphingomyelin were determined by gas chromatography-mass spectrometry in adrenoleukodystrophy (ALD) without adrenal insufficiency. Mass chromatogram tracing with the ion at m/z 143 [(CH2)6COOH3]+ showed increases of saturated very long chain fatty acids in plasma and erythrocyte membrane sphingomyelin in ALD. The C26:0/C22:0 ratios in plasma were 0.121, 0.057 and 0.007 in cases 1 and 2, and a control subject, respectively. The C26:0/C22:0 ratios in erythrocyte membrane sphingomyelin were 0.386, 0.211 and 0.093 in cases 1 and 2 and the control subject, respectively. The demyelinating process of ALD was clearly observed in both the inversion recovery 2100/500 and spin echo 2100/80 scans on magnetic resonance imaging. The magnetic resonance image in case 1 revealed widespread demyelinated lesions, involving almost the entire cerebrum and cerebellum, at 4 years after the onset, while that in case 2 revealed demyelinated lesions mainly limited to parieto-occipital areas at 1 year after the onset.

The presence of antibodies against adult T cell leukemia antigen (ATLA) was studied in 59 patients with chronic respiratory diseases. Of 13 patients with diffuse panbronchiolitis, 3, who developed adult T cell leukemia, had the anti-ATLA antibody and 8 had the related, anti-ATLA-like antibody. Of 13 cases of idiopathic interstitial pneumonia, 8 had the anti-ATLA-like antibody. Except for only one case, these antibodies were not detected in 33 patients with bronchial asthma or sarcoidosis and 20 healthy adults examined. These results suggested that the test of these antibodies would be useful for the diagnosis of diffuse panbronchiolitis and idiopathic interstitial pneumonia which frequently develop lung cancers.

Human leukemic B-cell (BALL-1), T-cell (TALL-1), and null-cell (NALL-1) lines were established from three patients with acute lymphoblastic leukemia (ALL). Intraperitoneal transplantation of these ALL cell lines into immunosuppressed newborn hamsters resulted in the development of invasive tumors in all recipients, except 4 of 10 implanted with NALL-1 line. BALL-1 and TALL-1 lines were serially transplanted for 10 and 9 passages, respectively. In addition to a common in vivo feature of mesenteric and retroperitoneal tumors, BALL-1 line was characterized by infiltration of the skin, massive ascites, and bone marrow invasion. TALL-1 cells infiltrated various organs including the lymph nodes, liver, bone marrow, central nervous system, eyes, etc. NALL-1 line grew slowly, producing the least number of tumors. Leukemic cells were detected in the blood of 24 of 25 hamsters bearing BALL-1 and in the blood of 18 of 24 hamsters bearing TALL-1. Thus, these three ALL cell lines were found to exhibit a characteristic biological behavior in hamsters.

Chromosome aberrations and clinical features of three patients with adult T cell leukemia-lymphoma (ATL) not associated with human T cell leukemia virus type I (HTLV-I) are described. From their clinical features, two patients were diagnosed as acute type and one patient was diagnosed as chronic type, which later converted to acute crisis. Clonal and many chromosomal abnormalities were observed before therapy in the two acute type cases and at relapse in the chronic type case. Karyotype aberrations, including trisomy 3, trisomy 7, trisomy 21, del(6)(q21), del(10)(p13), 14q11 translocation, and loss of X chromosome, all of which are frequently found in HTLV-I associated ATL, were also seen in these cases of HTLV-I-negative ATL.

Human leukemic B-cell (BALL-1), T-cell (TALL-1) and null-cell (NALL-1) lines have been established from three patients with acute lymphoblastic leukemia (ALL). To study the heterotransplantability and in vivo growth characteristics, attempts were made to transplant these ALL cell lines into newborn Syrian hamsters treated with rabbit anti-hamster thymocyte serum. Intraperitoneal implantation of 1.8-3.5 x 10(7) cells gave rise to invasive tumors in all recipients after 15 to 41 days. In addition to a common in vivo feature of mesenteric and retroperitoneal tumors, BALL-1 line was characterized by infiltration of the skin, massive ascites and bone marrow invasion. TALL-1 cells infiltrated various organs including the lymph nodes, liver, gallbladder, spleen, bone marrow, central nervous system and eyes. NALL-1 line grew slowly, producing the least tumors, although there were distant metastases in the lungs. Tumor cells were detected in the blood of 2 of 3 BALL-1-bearing hamsters and in the blood of 4 of 5 TALL-1-bearing hamsters. Thus, these three ALL cell lines were found to exhibit a characteristic biological behavior in hamsters, which might be related to the different cell lineage.

The chromosomes of a cell line (NALM-1) derived from the leukocytes of a patient with chronic myelocytic leukemia (CML) were examined with several banding techniques. The modal chromosome number was 46 and the cells contained a Philadelphia chromosome (Ph1), due to the standard translocation of the missing segment of the long arm of chromosome No. 22 onto the distal end of the long arm of chromosome No. 9, i.e., t(9;22) (q34;q11). The Ph1-positive modal cells of the NALM-1 line also had two common marker chromosomes, an extra X-chromosome, and missing chromosomes in groups No. 7, 9, and 15. Immunologic examination of the NALM-1 cells revealed them to have non-T-non-B (null) surface characteristics. An antigen specific for cells of acute leukemia and a human la-like antigen were detected. These facts suggested that the NALM-1 cell line originated from CML cells and maintained the cytogenetic and Immunologic characteristics of such cells.