Measurements of Strong-Interaction Effects in Kaonic-Helium Isotopes at Sub-eV Precision with X-Ray Microcalorimeters.

We have measured the 3d→2p transition x rays of kaonic ^{3}He and ^{4}He atoms using superconducting transition-edge-sensor microcalorimeters with an energy resolution better than 6 eV (FWHM). We determined the energies to be 6224.5±0.4(stat)±0.2(syst) eV and 6463.7±0.3(stat)±0.1(syst) eV, and widths to be 2.5±1.0(stat)±0.4(syst) eV and 1.0±0.6(stat)±0.3(stat) eV, for kaonic ^{3}He and ^{4}He, respectively. These values are nearly 10 times more precise than in previous measurements. Our results exclude the large strong-interaction shifts and widths that are suggested by a coupled-channel approach and agree with calculations based on optical-potential models.

Alu-Alu fusion sequences identified at junction sites of copy number amplified regions in cancer cell lines.

Alu elements are short, ∼300-bp stretches of DNA and are the most abundant repetitive elements in the human genome. A large number of chromosomal rearrangements mediated by Alu-Alu recombination have been reported in germline cells, but only a few in somatic cells. Cancer development is frequently accompanied by various chromosomal rearrangements including gene amplification. To explore an involvement of Alu-Alu fusion in gene amplification events, we determined 20 junction site sequences of 5 highly amplified regions in 4 cancer cell lines. The amplified regions exhibited a common copy number profile: a stair-like increase with multiple segments, which is implicated in the breakage-fusion-bridge (BFB) cycle-mediated amplification. All of the sequences determined were characterized as head-to-head or tail-to-tail fusion of sequences separated by 1-5 kb in the genome sequence. Of these, 4 junction site sequences were identified as Alu-Alu fusions between inverted, paired Alu elements with relatively long overlapping sequences of 17, 21, 22, and 24 bp. Together with genome mapping data of Alu elements, these findings suggest that when breakages occur at or near inverted, paired Alu elements in the process of BFB cycle-mediated amplification, sequence homology of Alu elements is frequently used to repair the broken ends.

Coamplification of multiple regions of chromosome 2, including MYCN, in a single patchwork amplicon in cancer cell lines.

Coamplification of multiple segments of chromosome 2, including an MYCN-bearing segment, was examined in 2 cancer cell lines, NCI-H69 (lung cancer) and IMR-32 (neuroblastoma). High-resolution array-CGH analysis revealed 13 and 6 highly amplified segments located at different sites in chromosome 2 in NCI-H69 and IMR-32, respectively. FISH analysis demonstrated that these segments were co-localized in double minutes in NCI-H69 and in homogeneously staining regions in IMR-32. Connectivity of the segments was determined by a PCR assay using designed primer sets. It was found that all the segments were connected to each other irrespective of their order and orientation against the genome sequence, and a single chain-like cluster was configured in both cell lines. Such patchwork structures of the amplicons suggest the possibility that massive genomic rearrangements, explained by the single catastrophic event model, are involved in the formation of the amplicons, enabling the coamplification of different chromosomal regions including the MYCN locus. The model comprises massive fragmentation of chromosomes and random rejoining of the fragments.

Subcellular fate of protein antibiotic neocarzinostatin in culture of a lymphoid cell line from Burkitt's lymphoma.

14C-LABELED PROTEIN ANTIBIOTIC NEOCARZINOSTATIN (NCS) was prepared efficiently by chemical modification. With the use of lymphoma-derived cell line P3HR-1, the subcellular behavior of this antitumor antibiotic was studied by the uptake a. nd autoradiography of isolated nuclei of radioactive NCS. The antibiotic was taken up by the cells, reaching the maximum value at 1.5 hr and decreasing in value at 4.0 hr to the level at 0.5 hr. The silver grains in the autoradiograms were also found in the isolated nuclei. The grain count in the nuclei showed a tendency similar to the uptake of NCS by the whole cells, i.e., a gradual increase at 0.5 hr, reaching the maximum value at 1.5 hr, and then decreasing after 4.0 hr to the level at 0.5 hr. These facts indicated that NCS reached not only to cytosol but also into the nucleus, and/or at least to the nuclear membrane of the lymphoid cell. The number of NCS molecules incorporated into the cells at 1.5 hr was calculated to be about 1 x 10-6/cells at a concentration of 3 mug NCS per ml of medium, which can be extrapolated to 1 x 10-4 molecules per cell at the minimum inhibitory concentration. The number of molecules should be even less within the nucleus. In cell-free systems, the interaction of DNA and NCS, which is an inhibitor of DNA synthesis, was investigated with use of a Sephadex G-100 column, with negative results. In the cell culture system, NCS molecules were degraded into smaller polypeptides of certain sizes by proteolysis either by serum component(s) or by cells themselves. An inactive isomer, pre-NCS, which is an antagonist of NCS and a partially denatured homologous molecule, behaved similarly to NCS in all of these experiments. Because the chemically modified NCS used in this study retained biological activity essentially similar to that of parental NCS, the results obtained here could be interpreted as similar to those of parental NCS in vitro.

Age-related changes in myelopoietic response to lipopolysaccharide in senescence-accelerated (SAM) mice.

The effects of in vivo lipopolysaccharide (LPS) administration on myelopoiesis were examined in senescence-accelerated (SAM) mice. Young mice injected with LPS exhibited: (a) increased femoral proliferative pool size; (b) transient reduction in femoral non-proliferative pool size and number of femoral colony forming unit-granulocyte macrophages (CFU-GMs); (c) marked increase in splenic CFU-GMs; and (d) transient increase in S-phase of femoral CFU-GMs. The responses of old mice after LPS administration differed from those of young mice in the following points: (a) no recovery of the femoral non-proliferative pool or femoral CFU-GMs, (b) less significant augmentation of the femoral proliferative pool and splenic CFU-GMs, and (c) prolonged reduction in S-phase of femoral CFU-GM. Injection of LPS into mice resulted in a hyperproduction of colony-stimulating activity (CSA) in bone followed by production of colony-inhibitory activity (CIA) in young mice and in contrast, an excessive CIA secretion from bone without an increase in CSA levels in old mice. These imbalances in the regulatory factors derived from non-hemopoietic cells in the bones may lead to an inappropriate response of myelopoiesis in aged SAM mice after LPS administration, which may play a key role in infections.

Comparison of erythropoietic response to androgen in young and old senescence accelerated mice.

In this study, to clarify whether the functional capacity of hemopoietic progenitor cells and the micro-environment of aged mice are identical with those of the young, we investigated the changes in the number of hemopoietic progenitor cells and the production of regulatory cytokines from splenic cells as well as changes in the serum levels of cytokine in senescence-accelerated mice (SAM) after administration of 19-nandrolone decanoate (19-ND), a synthetic androgenic anabolic steroid. 19-ND induced an increase in erythroid colony-forming units (CFU-E), erythroid burst-forming units (BFU-E), and granulocytic-macrophage committed progenitor cells (CFU-GM) in bone marrow and spleen; especially remarkable increases were observed in the splenic CFU-E in both young and old mice. Antigen expression analysis of hemopoietic organs revealed that total TER-119+ cells per spleen of young and old mice with androgen treatment rose 2.6- and 3.2-fold over their respective control values. The responsiveness of hemopoietic progenitor cells to androgen did not change with age. Injection of 19-ND into young and old mice markedly enhanced the erythropoietin levels but not IL3 and GM-CSF levels in the serum of both groups. Cytokine production assessed by pokeweed mitogen-stimulated spleen condition medium showed an age-related decline. Androgen treatment could not influence IL-3 and GM-CSF production of spleen. These findings suggest that the spleen of both old and young mice served as the major site of regenerative repopulation of hemopoietic progenitors, especially the late erythroid progenitors in 19-ND-treated mice. The proliferative reserve of erythropoiesis with androgen treatment in aged mice was not reduced more than that in treated-young mice.

Actin-actin contact: chemical cross-linking between actin and the 2.6-kDa peptide from subdomain 4 of actin.

Previously, we demonstrated that the 2.6-kDa peptide extending from Arg177 to Tyr198 in subdomain 4 of rabbit skeletal actin bound to actin itself, inhibited the elongation of actin filament, and severed F-actin. The corresponding segment in actin, therefore, is thought to contain the most critical actin-actin contact [Hori, K. and Morita, F. (1992) J. Biochem. 112, 401-408; Hori, K., Itoh, T., Takahashi, K., and Morita, F. (1994) Biochim. Biophys. Acta 1186, 35-42]. In this paper, we report on the binding site in actin for the 2.6-kDa peptide studied by using a zero-length cross-linker, 1-ethyl-3(3-dimethylaminopropyl)carbodiimide (EDC). We conducted limited digestion of actin cross-linked with the 125I-labeled 2.6-kDa peptide with various proteases, and developed peptide maps. The cross-linked region of the 2.6-kDa peptide was found to be within the region of Ala114 to Glu167 in actin by identifying the radioactive peptide fragments. The region was further restricted by isolation of radioactive peptide from alpha-chymotryptic digest of the cross-linked actin. The binding site of the 2.6-kDa peptide was finally assigned to be within the 24 amino acid segment from Ala144 to Glu167, which lies in subdomain 3 of actin. Using computer graphics, actin-actin contact provided by the two segments was suggested to be along the left-handed genetic helix of actin filament.

Molecular cloning of porcine neuronatin and analysis of its expression during pituitary ontogeny.

We cloned and sequenced two types (alpha and beta type) of cDNA from the porcine anterior pituitary cDNA library, encoding neuronatin that has been cloned as a gene expressed by the fetal developing brain. Nucleotide sequences of alpha and beta type were identical except for two gaps, suggesting that they were produced by an alternative splicing. The amino acid sequences showed a high homology (more than 94 %) compared to those of other mammals, including human, rat, and mouse. In this study, RT-PCR was performed for the RNA samples prepared from the porcine fetal and postnatal pituitaries. The results showed that two types of neuronatin are expressed through the fetal stages, from day 40 to day 110 in both sexes. The relative amounts of alpha to beta type reversed just after birth in both sexes, and both amounts increased further in the postnatal anterior pituitary. This increased expression after birth is quite different from the brain in which the expression of neuronatins decreased, indicating the distinct role of the neuronatin in pituitary and brain. Finally, we found a pituitary cell type specific localization, especially in gonadotroph cell lines LbetaT2, of beta-neuronatins.

Therapy-related megakaryoblastic leukemia with pituitary involvement following treatment for non-Hodgkin's lymphoma.

A case of a 66-year-old Japanese man developed therapy-related megakaryoblastic leukemia with pituitary involvement after chemotherapy for non-Hodgkin's lymphoma. Alkylating agents had been administered for the treatment of non-Hodgkin's lymphoma and 6 years later, megakaryoblastic leukemia with myelofibrosis and myelodysplasia developed. The blast cells expressed CD41, and immature antigens also. These findings were compatible with therapy-related megakaryoblastic leukemia. An autopsy revealed blast-cell infiltration into multiple organs including the posterior pituitary lobe. Therapy-related megakaryoblastic leukemia is very rare, and pituitary involvement may be associated with immaturity of blast cells.

[Plasma cell leukemia (IgG kappa) presenting bilateral neurosensory hearing loss and left sixth cranial nerve plasy].

A 30-year-old man who had been given a diagnosis of IgG-kappa multiple myeloma by another hospital and treated with melphalan, prednisone, and cyclophosphamide 6 months earlier, was admitted to our hospitaly in July 1994 because of progressively impaired hearing in both ears, vertigo, and worsening fatigue. Peripheral blood examination showed a white blood cell count 25,000/microliter, with 77.5% atypical plasma cells. Examination at the time of hospitalization also revealed retinal hemorrhages and serum hyperviscosity. The diagnosis was plasma cell leukemia with hyperviscosity syndrome. Subsequent treatment consisted of vincristine, doxorubicine, and prednisone and repeated plasmapheresis. This resulted in a partial response and a reduction of serum viscosity but no reversal of hearing loss. One month after admission, left sixth cranial nerve plasy was demonstrated. Cranial computed tomography studies disclosed a tumoral mass in the sphenoid sinus. The patient received local radiotherapy and intensive chemotherapy, but exhibited no notable alleviation of his cranial nerve palsy. He died of septicemia and progressive disease in August 1994. This case was rare in that it involved plasma cell leukemia and bilateral neurosensory hearing loss associated with serum hyperviscosity and sixth cranial nerve plasy due to plasmacytoma within the sphenoid sinus.

[Plasma cell leukemia presenting with circulating villous lymphocytes and an indolent clinical course].

A 73-year-old man was admitted to our hospital in July 1996 because of lymphoctyosis and lumbago. Physical examination revealed hepatomegaly and anemia. Hematologic examination showed a hemoglobin concentration of 9.6 g/dl and a leukocyte count of 32,700/microliter with 74% abnormal mononuclear cells. In Wright-Giemsa stained blood films, these cells had short villi arising from 1 or 2 poles. Immunophenotyping of peripheral mononuclear cells showed moderate to strong expression of CD10, CD24, CD38, and sIg lambda, but not of CD19, CD20, or CD25. Southern blot analysis of the peripheral mononuclear cells demonstrated rearranged monoclonal bands in the C lambda. Urine immunoelectrophoresis detected a monoclonal band identifiable as lambda-type Bence Jones protein. In addition, bone X-ray studies disclosed multiple osteolytic lesions. A diagnosis of plasma cell leukemia was made, and the patient was placed on chemotherapy consisting of cyclophosphamide and prednisolone. No notable improvement in laboratory findings was seen but the patient experienced an indolent clinical course. He died of pneumonia in January 1998. The morphological and clinical findings were unusual for a case of plasma cell leukemia. This case study suggested that signs of lymphocytosis require immunophenotypic and electron microscopic studies for the differential diagnosis of plasma cell leukemia.

[A case of chronic myelogenous leukemia presenting multiple extramedullary tumors localized in cranial dura].

A 64-year-old woman had been given a diagnosis of Ph-positive chronic myelogenous leukemia (Ph+ CML) in October 1992 and accordingly treated with interferon-alpha busulfan, and hydroxyurea. She was admitted to our hospital with a one-day history of consciousness disturbance on May 30, 1993. Two weeks before admission, she had received chemotherapy consisting of vincristine and predonisolone because of progressive thrombocytopenia, basophilia, and leukocytosis accompanied by a heightened degree of cell immaturity in peripheral blood and bone marrow. Cranial computerized tomography on admission disclosed tumoral masses in the left frontal lobe and the right temporal lobe. Moreover, lumbar puncture ezinkns disclosed blastoid cells in cerebrospinal fluid. Based on these laboratory findings, the diagnosis was blastic crisis CML, 46XX t(9; 22; 17) (q34; q11; q23), cytogenetic aberration and extramedulary brain disease Although the patient underwent the same combined chemotherapy again, her unconsciousness did not resolve. She died of cerebellar herniation on the 7th hospital day. Post mortem examination revealed three extramedullary tumors localized in cranial dura. This was a rare case of CML presenting multiple extramedullary tumors localized in cranial dura.

[Severe hemolytic anemia with tear drop red cells as initial manifestation of Wilson's disease].

A 16-year-old girl was admitted for a detailed examination of hemolytic anemia in November 1995. Initial laboratory findings included a total bilirubin concentration of 1.46 mg/dl, hemoglobin of 9.1 g/dl, and a reticulocyte count of 89/1000 percent. The plasma haptoglobin concentration was below 10 mg/dl. A blood smear showed many dacryocytes and a few echinocytes and codocytes. GOT was 71 IU/l; GPT, 44 IU/l; and LDH, 812 IU/l; the results of a hepaplastin test were 45% of normal. On further investigation, the level of serum ceruloplasmin was found to be 4 mg/dl, and of serum copper, 43 micrograms/dl. Urinary copper excretion was markedly increased, at 345 micrograms per day. Slit-lamp examination of both corneas revealed obvious Kayser-Fleischer rings. A liver biopsy sample showed fibrosis histologically and an elevated copper concentration of 535 micrograms/g dry weight and 183 micrograms/g wet weight. In family studies, the patient's asymptomatic 5-year-old sister was observed to have metabolic abnormalities consistent with Wilson's disease. These findings suggested that the patient's hemolytic anemia with red cell deformities was due to abnormal copper metabolism associated with Wilson's disease.

[Surgical treatment in an infant with atrial septal defect and persistent left superior vena cava draining into the coronary sinus; report of a case].

A 8-month-old boy with ostium secundum atrial septal defect and persistent left superior vena cava draining into the coronary sinus underwent surgical treatment because he had required long-term ventilator support due to congestive heart failure and respiratory distress. The left superior vena cava and dilated coronary sinus produce a ridge in the left atrium, causing a subdivided left atrium. In this case, the early symptom could be explained by an obligatory left to right shunt at the level of the left atrium resulting from a ridge produced by the left superior vena cava and the colonary sinus, as a subdivided left atrium.

[Management of mediastinitis after pediatric cardiac surgery with continuous closed irrigation method].

Two cases are presented as a successful management for mediastinitis with the continuous closed irrigation method after pediatric cardiac surgery. The continuous closed irrigation method has significant advantages over conventional open irrigation method or muscle flap in pediatric cases, because the system is simple to handle and easy to wash out any infectious tissue debris without additional invasive intervention. However, adequate duration of the irrigation and subsequent antibiotic regimen is still unclear. We conclude that the continuous closed irrigation method is an effective management which is applicable to most of mediastinitis cases after pediatric cardiac surgery.

[Pollen information of airborne Japanese cedar pollen using a simulation method].

We have developed a simulation method of airborne Cryptomeria japonica pollen distribution on a map displayed visually on a TV screen. Each patient can be available the information where the place he or she lives. The pollen season in 1995, we served the information about airborne pollen distribution on a map and C. japonica flowering areas on a map to a local resident through TV broadcasting. To verify the simulation method, comparison was made between the result from actual pollen counting and from simulation. It was clarified that both results were comparatively agreed on daily basis. Problem about compatibility among personal computers were solved to rewrite the program of displaying the image using Visual Basic for MS-Windows and create image files. The files can be read continuously by animation software. We think the information can be offered to local resident, local clinicians and patients waiting at the clinics by use of computer networks.

[Effects of global climate change on Japanese cedar pollen concentration in air--estimated results obtained from Yamagata City and its surrounding area].

The effects of global warming on Japanese cedar pollen concentration in air were examined. There was a significant increase in total pollen count in years where small or average values of pollen dispersion were observed and where summertime temperatures were postulated to have risen from 2 to 5 degrees C. There were no remarkable differences in total pollen count in years where large values of pollen dispersion were observed. On the other hand, a drastic decrease in total pollen count was expected when summertime temperatures were postulated to have dropped from 2 to 5 degrees C. A major factor controlled the value of total pollen count was examined by use of a simulation method of airborne Cryptomeria japonica pollen. The values of total pollen count mainly controlled by the amount of male flower at basins and in Japanese cedar forested areas, and the manner in which plains isolated from pollen sources are considered in part to be subjected to meteorological conditions. The difference of initiation time of the pollen season has no great influence on the values of total pollen count.

[An attempt of applying the image processing for the automatic estimation of sampled airborne pollen].

Development of techniques to rapidly and easily estimate an airborne pollen quantity is necessitated in order to make out a appropriate prescription for an allergy patient from medical clinical viewpoints, and in order to research a movement of allergen from medical basic viewpoint. The measurement of airborne pollen quantity required a large labour and time, because the amount of pollen grains is visually measured by naked eye. This paper, as a first step to estimate an airborne pollen quantity full-automatically, discuss techniques to measure the quantity of sampled airborne cedar pollen automatically and rapidly using the image processing techniques. As a result, the following facts are cleared. Automatic measurement is possible to some degree without any special image processing. It is important to eliminate a noise on image as blurs on basefilm for high accuracy measurement. The precision has improved fairly with level correction for image. Sharpening filter is the most appropriate process to improve the accuracy of automatic measurement of sampled airborne pollen. This filtering process has a merit that the operativeness is easy.

[Measurement of pain threshold and tolerance threshold using radiant heat stimulation.--Regional difference, sex difference, expression of sensation].

In 1940, Hardy had developed a method of radiant heat stimulation and succeeded to determine pain threshold quantitatively. An algometer NYT-55 was developed according to his principle by Nakahama and Yamamoto, which was a further developed version of Hardy's model. In the present study, pain threshold and tolerance threshold were measured quantitatively, using Initial Skin Temperature Controlled Repeated Thermal Stimulation System (STRTS) which was composed of NYT-55 (KUDO ELECTRIC Co.) and a microcomputer PC-9801 VM 21 (NEC Co.). The following results were obtained in healthy volunteer students (62 males and 61 females). The pain and tolerance threshold were changed at every measurement sites on the body surface. The sex differences were observed for both pain and tolerance threshold. In general, the male's threshold intensity was higher than that of female. The pain threshold of trained subjects who have experienced preparatory training about pain measurement were relatively stable, while the threshold of untrained subjects were unstable.