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Two-photon imaging is a major recording technique in neuroscience but its low sampling rate imposes a severe limit of elucidating high temporal profiles of neuronal dynamics. Here we developed two hyperacuity Bayesian algorithms to improve spike detection and spike time precision, minimizing the estimation error supervised by the ground-truth given as the electrical spike signals. The benchmark showed that our algorithms outperformed other unsupervised algorithms maximizing the likelihood of the estimates for both experimental and simulation data. We argue that the supervised algorithms are useful tools to improve spike estimation of two-photon recording in case ground truth signals are available.

BACKGROUNDS AND PURPOSE Lactic acidosis is a fatal adverse effect of metformin, but the risk factor remains unclear. Multidrug and toxin extrusion 1 (MATE1) is expressed in the luminal membrane of the kidney and liver. MATE1 was revealed to be responsible for the tubular and biliary secretion of metformin. Therefore, some MATE polymorphisms, that cause it to function abnormally, are hypothesized to induce lactic acidosis. The purpose of this study is to clarify the association between MATE dysfunction and metformin-induced lactic acidosis. EXPERIMENTAL APPROACH Blood lactate, pH and bicarbonate ion (HCO3-) levels were evaluated during continuous administration of 3 mg·mL−1 metformin in drinking water using Mate1 knockout (−/−), heterozygous (+/−) and wild-type (+/+) mice. To determine the tissue accumulation of metformin, mice were given 400 mg·kg−1 metformin orally. Furthermore, blood lactate data were obtained from diabetic patients given metformin. KEY RESULTS Seven days after metformin administration in drinking water, significantly higher blood lactate, lower pH and HCO3- levels were observed in Mate1−/− mice, but not in Mate1+/− mice. The blood lactate levels were not affected in patients with the heterozygous MATE variant (MATE1-L125F, MATE1-G64D, MATE2-K-G211V). Sixty minutes after metformin administration (400 mg·kg−1, p.o.) the hepatic concentration of metformin was markedly higher in Mate1−/− mice than in Mate1+/+ mice. CONCLUSION AND IMPLICATIONS MATE1 dysfunction caused a marked elevation in the metformin concentration in the liver and led to lactic acidosis, suggesting that the homozygous MATE1 variant could be one of the risk factors for metformin-induced lactic acidosis.

This paper describes results obtained from analysis and experiments done on a fluxgate dc earth leakage current sensor for electric vehicles and photovoltaic power generation systems. As fluxgate current sensors are extremely sensitive to room temperature, various applications need this characteristic. However, conventional fluxgate current sensors have a problem in that they can only measure a narrow range of currents. To address this issue, we propose that the turns of the excitation coil be optimized with a simulator. This new sensor consists of a coiled wire that is wound around a toroidal amorphous core and through a bus bar. The sensor detects variations in excitation current according to dc current. The sensor was analyzed with a simulator. The magnetic field simulator was used to calculate the excitation current, and the output voltage according to the excitation current was calculated with a circuit simulator. We found that the results from simulations almost corresponded to those from measurements, and were able to verify the validity of the basic analytical model. As a result of analysis with the simulator, 35 turns of the excitation coil were used to achieve a wide range of measurements of ±600 mA.

In order to investigate the crack path under Mode II or Mode III loadings, reversed torsion tests were carried out on SAE52100 and Mode II fatigue crack growth tests were carried out on 0.47% carbon steel specimens. In the torsional fatigue test (SAE52100), the type of inclusion at the torsional fatigue fracture origin was slender MnS inclusions which were elongated in the longitudinal direction. The cracks first propagated by Mode II up to crack length 2a = 100–200 μm (which are almost equal to the length of MnS inclusion) in the longitudinal direction, and then branched by Mode I to the direction (∼ ± 70.5°) perpendicular to the local maximum normal stress (σθmax) at the crack tip. In the Mode II fatigue crack growth test (0.47% carbon steel) in air and in a vacuum, the cracks first propagated by Mode II. After the Mode II fatigue crack growth stopped, the crack branched to the direction perpendicular to the local maximum normal stress (σθmax) at the crack tip, and finally branched to the angle close to the direction perpendicular to the remote maximum principal stresses. A fibrous pattern on the Mode II fatigue fracture surface tested in a vacuum was clearer than that in air. The Mode II threshold stress intensity factor ranges, ΔKIIth = 10.2 MPa m (Longitudinal crack) and ΔKIIth = 12.5 MPa m (Transverse crack) in a vacuum were higher than those in air, ΔKIIth = 9.4 MPa m (Longitudinal crack) and ΔKIIth = 10.8 MPa m (Transverse crack). Both in vacuum and in air, the values of ΔKIIth for crack growth perpendicular to the rolling direction were higher than those for crack growth parallel to the rolling direction. The values of KII and KIII at a 3D elliptical crack tip under shear stress were analyzed to investigate the shear crack growth pattern in materials. The 3D crack analysis shows that the most stable aspect ratio b/a of a small planar elliptical crack under cyclic shear stress is b/a = 0.49 in absence of friction on the crack surfaces. The aspect ratio b/a = 0.49 can be explained by the equal resistance against fatigue crack growth both in Mode II and Mode III, i.e., ΔKIIth = ΔKIIIth. However, the aspect ratio b/a for the failure of a real railway wheel did not stay at the stable aspect ratio b/a = 0.49 and continued decreasing. The cause for the decrease in the aspect ratio b/a smaller than 0.49 was revealed to be the friction between crack surfaces.

In order to investigate the crack path in materials containing a small crack under mixed mode loading, reversed torsion and combined push-pull/torsion fatigue tests were carried out on 0.47% carbon steel specimens containing an initial small crack. The initial small cracks were introduced by a preliminary push-pull fatigue test using a specimen that contained an artificial small hole of 40 μm diameter/depth. Push-pull fatigue tests followed by reversed torsion, and reversed torsion followed by push-pull were carried out. Fatigue tests of combined push-pull/torsion followed by push-pull were also carried out to examine the effect of crack geometry, such as branching and kinking, on cumulative fatigue damage. Different crack growth behaviours due to different loading modes and sequences complicatedly influenced the fatigue crack path and eventually the cumulative fatigue damage. Thus, existing fatigue damage theories cannot be applied to the cases presented in this study. A single factory roof morphology was formed on the fracture surface of the specimen having an initial semielliptical surface crack under cyclic torsion. Taking the result into consideration, torsional fatigue tests of circumferentially cracked specimens were carried out to investigate the Mode III crack growth threshold and the formation mechanism of the factory roof. It was found that the factory-roof pattern was formed by Mode I crack branching from small semielliptical cracks nucleated ahead of the circumferential crack tip. Thus, stress intensity factors at the semielliptical crack tip ahead of the circumferential crack tip were analysed to make the mechanism of crack branching from the semielliptical crack clear. The analysis shows that the Mode III stress intensity factor, K III , at the deepest point of the semielliptical crack decreases with increase in the depth of semielliptical crack. It follows that as the shear crack growth becomes difficult at the semielliptical crack tip, crack branching starts at the major axis ends in the direction for Mode I growth, forming factory roof, due to ΔK Ith < ΔK IIIth .

The central nervous system of persons with Down syndrome presents cytoarchitectural abnormalities that likely result from gene-dosage effects affecting the expression of key developmental genes. To test this hypothesis, we have investigated the transcriptome of the cerebellum of the Ts1Cje mouse model of Down syndrome during postnatal development using microarrays and quantitative PCR (qPCR). Genes present in three copies were consistently overexpressed, with a mean ratio relative to euploid of 1.52 as determined by qPCR. Out of 63 three-copy genes tested, only five, nine and seven genes had ratios >2 or

Quantitative crossed cerebellar diaschisis (CCD) and the correlation with a reduction in supratentorial regional cerebral blood flow (rCBF) and cerebrovascular reserve capacity (CVR) were investigated in clinically stable patients with major cerebral artery occlusion by the iodine-123-N-isopropyl-p-iodoamphetamine (I-123 IMP) single photon emission computed tomography (SPECT) method. Thirty patients with major cerebral artery occlusion underwent SPECT by the I-123 IMP autoradiographic method. Regional CBF was measured in the cerebral hemisphere, frontal and parietal lobes, temporo-parietal lobe, and cerebellum both at rest and after administration of acetazolamide. Eighteen of 30 patients (60%) had CCD. CCD was significantly related to magnetic resonance imaging evidence of infarction. Quantitative CCD was 17% and the CVR in the cerebellum was preserved in patients with CCD. There was a significant difference in CBF and CVR between the affected and normal sides in all regions of interest in the patients without CCD [CBF (ml/100 g/min): hemisphere (H), normal side (N): 31.4 +/- 6.8, affected side (A): 27.5 +/- 7.4; p0.05. CVR: H, N: 0.56 +/- 0.38, A: 0.42 +/- 0.18; p0.01]. CCD is common in patients with major cerebral artery occlusion, and quantitative I-123 IMP SPECT is helpful in detecting CCD in clinically stable patients with occlusion of major cerebral arteries.

Investigates the analysis of a hyperthermic treatment. The time dependent electromagnetic-heat transfer equations are solved step-by-step considering the temperature dependencies of relative permittivity /spl epsiv//sub r/ and electric conductivity /spl sigma/. Close agreement between the calculated and measured temperature distributions is obtained. A technique to direct the electromagnetic energy toward a specific part of a homogeneous torso model is also noted.

The stereoselective deuterium labeling at the 5' methylene protons of the ribose ring recently developed by Kawashima et al. [1995, Tetrahedron Lett., 36, 6699-6700] enabled the assignment of pro-R and pro-S protons at the 5' position. The deuterium-labeled nucleotides, [(5'S)-(2)H]- and [(5'R)-(2)H]-diastereomers, in an approximate ratio of 2:1, were incorporated in the decamer 5'-d(GCATTAATGC)-3'. Thus, both pro-R and pro-S methylene proton signals without geminal coupling appeared in the NOESY and DQF-COSY spectra. Complete stereospecific assignments and simplified spin systems enabled the determination of 15 (3)J coupling constants between H4' and H5'/H5", and the unambiguous assignment of 135 NOESY cross peaks originating from H4'/H5'/H5" resonances.

A 31-year-old male having familial amyloid polyneuropathy underwent a Tc-99m(V)-DMSA study to evaluate the myocardial involvement. The patient also underwent T1-201, I-123-BMIPP and I-123-MIBG myocardial SPECT studies to evaluate blood perfusion, fatty acid metabolism and sympathetic function of the heart, respectively. Tc-99m(V)-DMSA SPECT showed uptake to the myocardium indicating myocardial involvement of amyloidosis. Both T1-201 and I-123-BMIPP studies showed normal uptake indicating normal blood perfusion and fatty acid metabolism but I-123-MIBG SPECT showed no uptake to the heart, indicating severe impairment of sympathetic function.

Haplotype analysis is important for mapping traits. Recently, methods for estimating haplotype frequencies from genotypes of unrelated individuals based on the expectation-maximization (EM) algorithm have been developed. Our program estimates haplotype frequencies in the population and determines the posterior probability distribution of diplotype configuration (diplotype distribution) for each subject based on the estimated haplotype frequencies. Samples from three ethnic groups for the smoothelin gene (SMTN) and those from three Japanese groups for serum amyloid A genes (SAA@) were analyzed. The estimated diplotype distribution for each individual was concentrated, in most cases, in a single diplotype configuration. The diplotype configuration thus determined was the same as that determined in in vitro experiments, with one exception. Thus, the diplotype configurations determined using the estimated haplotype frequencies from unrelated individuals are reliable. Using this method, the risk of a subject developing a phenotype may be estimated from the diplotype distribution when the phenotype is associated with diplotype configurations.

The mechanisms of uptake and intracellular distribution of 99mTc-tetrofosmin, 99mTc-MIBI and 201TI and the behaviors of 99mTc-tetrofosmin and 99mTc-MIBI in relation to Na+ were studied with primary cultures of myocardial cells.Both the uptake and the washout of the tracers were sequentially measured. The cells were treated with ouabain, bumetanide, tetrodotoxin, dimethyl amiloride (DMA), nigericin and carbonyl cyanide m-chlorophenylhydrazone (CCCP) to observe the effects of the uptake and intracellular distribution of the traders. Cells equilibrated in buffers with or without Na+ were treated with monensin and DMA to evaluate the effect of Na+ on the accumulation of the tracers.Despite the similarities in uptake kinetics, there was a higher level of retention of 99mTc-tetrofosmin inside the cells. Ouabain, bumetanide and tetrodotoxin did not show any inhibitory effect on the uptake of 99mTc-tetrofosmin and 99mTc-MIBI, whereas they produced various degrees of inhibition of 201TI uptake. DMA produced approximately 35% inhibition of 99mTc-tetrofosmin uptake and 50% inhibition of 99mTc-MIBI uptake. Nigericin increased the uptake of 99mTc-MIBI by the cells. The addition of CCCP produced the release of 38% of the accumulated 99mTc-tetrofosmin and 52%-70% of the accumulated 99mTc-MIBI, indicating that these percentages of accumulation were related to mitochondrial uptake. Neither Na+-free buffer nor monensin had any significant effect on 99mTc-tetrofosmin accumulation, but they both caused increased accumulation of 99mTc-MIBI.The uptake of both 99mTc-tetrofosmin and 99mTc-MIBI through the cell membrane is partly related to the Na+/H+ antiporter system. Only part of the accumulated 99mTc-tetrofosmin inside the cells enters into mitochondria; most of the accumulated 99mc-MIBI is related to mitochondrial uptake. This uptake may be related to Na+.

Substance P (SP) is a neuropeptide widely distributed in the nervous system. Extensive study has shown SP stimulates production of various cytokines by bone marrow stromal cells, although, the role of SP in hematopoietic phenomena is still unclear. Recently, we established a human cloned stromal cell line, HAS303, which can support hematopoietic stem cell proliferation and differentiation in vitro. We used this culture system to examine the effects of SP. Expression of the mRNAs of neurokinin (NK)-1R, NK-2R and NK-3R, specific SP receptors, on HAS303 cells was demonstrated by the RT-PCR. CD34+ cells isolated from bone marrow were co-cultivated with HAS303 cells in the presence and absence of SP and the total hematopoietic cells and progenitors were counted every 5 days. Introducing SP (10(-8) M) to the co-cultures significantly increased the number of total cells and progenitors compared with control cultures. SP showed no enhancing activity on CD34+ cells cultured alone. SP also stimulated IL-3-dependent colony formation of whole bone marrow MNCs in a soft agar culture system, but showed no such activity on isolated CD34+ cells in this system. These observations suggest that SP stimulated HAS303 cells, activated HAS303 cells, and stimulated the proliferation and differentiation of CD34+ cells. Treating HAS303 cells with SP increased the intracellular Ca2+ concentration and stimulated production of G-CSF, GM-CSF, SCF and IL-6, but not IL-1alpha, IL-1beta and TNF-alpha, but did not enhance proliferation. All these findings suggest that SP mediates hematopoietic cell proliferation and differentiation in vitro by activating stromal cell function.

A previously reported highly sensitive assay for measuring telomerase activity on cell and tissue extracts indicates that most human tumor tissues, but not cells adjacent to tumors, have detectable telomerase activity. Although this assay has provided a significant amount of information about the presence or absence of telomerase activity, it does not indicate whether all cells within a tumor have telomerase activity or whether only a subset does. The present report demonstrates the ability to advance this technology to an in situ assay. Using fluorescent telomerase primers and in situ PCR, we show that telomerase activity can be detected at the cellular level. This study demonstrates that telomerase activity is not detected in normal cells but is detected in tumor cells of clinical specimens and in tumor-derived cell lines.

We investigated and compared findings on combined 99mTc pertechnetate-201Tl with those of [99mTc]pertechnetate-67Ga scintiscans to elucidate the advantages of 201Tl in detecting various salivary glands disorders.We studied 23 patients: 6 had sialadenitis, 12 had benign tumors and 5 had malignant tumors. All but four patients had undergone [99mTc]pertechnetate (before and after lemon stimulation), 201Tl (early and delayed) and 67Ga imaging.Five of six sialadenitis patients showed various degrees of diffuse uptake of 99mTc. All six except one showed early uptake without retention of 201Tl on delayed imaging. The 67Ga scan showed uptake in all patients except one. Nine of 12 benign tumors showed a cold defect on 99mTc scans. Patients with Warthin's tumors and plasmacytoma showed increased 99mTc uptake at the tumor with retention. The 201Tl scan showed early uptake without retention in benign tumors except in three patients, two of whom had Warthin's tumor. Five of the benign tumors, however, were positive on 67Ga scan. None of the malignant tumors showed any uptake of 99mTc. The 201Tl scan showed uptake with tumor retention on delayed images in three patients; three other patients also had positive 67Ga scans. Overall, sensitivity and specificity of 201Tl in detecting malignant tumors were 60% and 73%, respectively, with a negative predictive value of 85%. Sensitivity and specificity for 67Ga were 60% and 47%, respectively, with a negative predictive value of 80%.In view of sensitivity, specificity and convenience of 201Tl as well as future prospects for dual-isotope acquisition, 67Ga may be replaced by 201Tl in detecting salivary gland disorder.

Genetic alteration, including genomic instability, is an ultimate step toward the malignant process. One approach to delineating replication errors in cancer cells is to determine the alterations of microsatellites, which are short, repeated nucleotide sequences existing throughout the genomes. We used a fluorescent system to assess microsatellite changes in seven loci (D2S123, D3S643, D5S107, LPL, D17S261, TP53, and D18S34) of 73 consecutive patients with various hematological neoplasias. De novo acute leukemia patients had a low frequency (1%) of microsatellite alterations at each locus, and none of them demonstrated multiple microsatellite changes. In chronic myeloid leukemia patients, no microsatellite instability was detected in the chronic phase, whereas a relatively high frequency (25%) of multiple microsatellite changes was evident in the blastic phase, and half of these patients had multiple microsatellite changes. About 50% of the patients with myelodysplastic syndrome (MDS) and post-MDS acute myeloid leukemia (post-MDS AML) had microsatellite alterations. We next compared microsatellite alterations in two different hematological phases (MDS and post-MDS AML phases); 5 of 11 patients with post-MDS AML had de novo appearance of microsatellite instability during disease progression. This indicates that genomic instability at multiple microsatellite loci could occur either before or after leukemic transformation in MDS patients. We concluded that genomic instability in chronic myeloid leukemia might be linked to blastic transformation in combination with cytogenetic changes. In contrast, MDS patients had replication errors as a relatively early genetic event as well as a late genetic event. These results suggest that the involvement of genomic instability in the progression of disease is different among various types of leukemia.

We investigated the kinetics, cellular uptake and intracellular distribution of 99mTc-tetrofosmin in tumor cell lines and compared them with those of 99mTc-MIBI and 201TI.At specific intervals after incubation with radiotracers, cellular uptake was determined. Cells were also treated with nigericin, carbonyl cyanide m-chloro-phenylhydrazone (CCCP) and ouabain to determine their effects on the uptake of the tracers.Each tracer showed similar uptake kinetics in both cell lines, and a steady-state was maintained for at least 4 hr. Nigericin stimulated the uptake of both 99mTc-tetrofosmin and 99mTc-MIBI in HBL-2 cells, although it inhibited their uptake in SW-13 cells. Nigericin also inhibited 90% of 201TI uptake in both cell lines. Addition of CCCP caused 73%-97% release of accumulated 99mTc-MIBI from both cell lines with or without nigericin pretreatment, indicating that most of the accumulated 99mTc-MIBI was related to mitochondria. The effect of CCCP on accumulated 99mTc-tetrofosmin was less marked than that on 99mTc-MIBI in both cell lines, indicating that only a part of accumulated 99mTc-tetrofosmin, was related to mitochondria. Ouabain preincubation inhibited 74%-77% and 51%-53% of 201TI uptake in HBL-2 and SW-13 cells, respectively, as well as inhibited 22%-31% uptake of 99mTc-tetrofosmin in both HBL-2 and SW-13 cells. Uptake by the dead cells of either cell line was negligible for each tracer.Technetium-99m-tetrofosmin uptake depends on both cell membrane and mitochondrial potentials. Only a small fraction of 99mTc-tetrofosmin accumulates inside the mitochondria, while most 99mTc-MIBI accumulates inside the mitochondria. Thallium-201 uptake is partly independent of the Na+, K+ pump.

Erythropoietin (Epo), the primary in vivo stimulator of erythroid proliferation and differentiation, acts, in part, by altering the tyrosine phosphorylation levels of various intracellular signaling molecules. These phosphorylation levels are tightly regulated by both tyrosine kinases and tyrosine phosphatases. We have recently shown that the SH2 containing tyrosine phosphatase, Syp, binds directly to both the tyrosine phosphorylated form of the Epo receptor (EpoR) and to Grb2 after Epo stimulation of M07e cells engineered to express high levels of human EpoRs (T. Tauchi, et al: J Biol Chem 270:5631, 1995). To determine which tyrosine within the EpoR is responsible for binding Syp, we examined DA-3 cell lines expressing full-length mutant EpoRs bearing tyrosine to phenylalanine substitutions for each of the eight tyrosines within the intracellular domain of the EpoR. We found that: (1) all Epo-stimulated mutant EpoRs, except for the Y425F EpoR, coimmunoprecipitated with Syp; (2) all Epo-stimulated mutant EpoRs, except for the Y425F EpoR, bound to a GST-fusion protein containing both SH2 domains of Syp; (3) Jak2 could phosphorylate GST-Syp in vitro after Epo stimulation of wild-type (wt) EpoR expressing DA-3 cells; (4) Epo-stimulated tyrosine phosphorylation of Syp in vivo was markedly reduced in Y425F EpoR expressing DA-3 calls; and (5) DA-3 cells expressing the Y425F EpoR grow less well in response to Epo than wt EpoR expressing cells. These results suggest that Syp binds via its SH2 domains to phosphorylated Y425 within the EpoR and is then phosphorylated on tyrosine residues by Jak2. Moreover, Y425 in the EpoR reduces the Epo requirement for Syp tyrosine phosphorylation and promotes proliferation.

We identified the telomere length at different hematological phases in 16 patients with myelodysplastic syndromes (MDS), showing disease evolution with a conventional Southern blot hybridization using the (TTAGGG)4 probe. The MDS patients studied were classified into three groups according to the pattern of telomere length reduction. The first group had telomere shortening at the time of disease diagnosis. In four of the six MDS patients in this group, the disease progressed within 6 months postdiagnosis and each of them survived for less than 1 year. Moreover, in this group four patients showed a 5q anomaly with or without additional changes, and 50% of patients in this group had complex chromosome abnormalities. The patients in the second group showed reductions in telomere length after disease progression; two of these three patients showed gradual disease progression and had one or two chromosome abnormalities. The third group comprised the remaining seven MDS patients; they showed no telomere reduction by disease evolution. Two patients in this group experienced rapid disease progression. These results may indicate that telomere reduction is linked to disease evolution in some MDS patients, perhaps as a result of genomic instability because patients with complex chromosome abnormalities were clustered in the first group. However, because some MDS patients show disease progression without telomere reduction, genetic changes, including point mutations of certain gene(s), may also contribute to disease progression. We further noted that telomere shortening at the time of MDS diagnosis might indicate a poor MDS prognosis.

Erythropoietin is known to be effective for the treatment of anemia in chronic renal failure, but the efficacy of erythropoietin for anemia in other diseases is not so great. Insulin exerts a growth promoting activity in various kinds of cells. In the present study, the effects of insulin on erythroid progenitors (colony forming units-erythroid, CFU-E; and burst forming units-erythroid, BFU-E) in human bone marrow were examined at various concentrations of recombinant human erythropoietin (rh-Epo) to clarify the relationship between erythropoietin and insulin. Human insulin stimulated the formation of CFU-E and BFU-E in the presence of three concentrations (0.25, 5, and 100 U/ml) of rh-Epo. Stimulatory effects of human insulin on CFU-E and BFU-E were also observed in the nonphagocytic and nonadherent bone marrow fraction (NP-NA fraction) and in the NP-NA and T cell-depleted fraction at each concentration of rh-Epo. Human insulin further stimulated the CFU-E and BFU-E growth in CD34+ separated bone marrow cells. These results indicate that the enhancing effect of human insulin on erythroid progenitors is not mediated through monocytes and macrophages or T cells, suggesting a direct action on erythroid progenitors.

Nagoya University Hospital has developed a new, comprehensive computerized hospital administration system since April 1992. In the present study, a user's evaluation questionnaire survey on the order entry system revealed that the improvements at the human interface level were appreciated by some respondents, but not all users felt the human interface provided, adequately meets their needs. Another presumed advantage of the micro-mainframe-link architecture is fast response, but in our study respondents considered the response time too long. Most users felt that transmission of orders was fast, accurate and clear, and that fast retrieval of the results of laboratory tests was good. Our system with this kind of architecture has thus proved satisfactory except for the slow response time. This study will be useful to reengineer our hospital information system in the next phase.

The blast progenitors in acute myelogenous leukemia grow in response to hematopoietic growth factors (HGFs), and their sensitivity to antileukemic drugs is influenced by HGFs. We report the effects of stem cell factor (SCF) on the growth and sensitivity to 1-beta-D-arabinofuranosylcytosine (Ara-C) of blast progenitors in acute myelogenous leukemia. SCF stimulated both colony formation and self-renewal of blast progenitors and, when used in combination with other HGFs, synergistic enhancement of colony formation was noted in 8 of the 15 patients examined. Cell fractionation studies demonstrated no unique growth dependency on SCF in either CD34+ or CD34- populations. Blast cells of patients that displayed synergistic growth enhancement with SCF displayed the highest Ara-C sensitivity when HGFs were used in combination with SCF. The tritiated thymidine suicide test (20-min exposure) revealed that the proportion of blast progenitors in the S phase of the cell cycle was highest when SCF and another HGF were simultaneously present, although 24-h exposure killed most or all of the blast progenitors. These data indicate that SCF enhances growth and sensitivity to Ara-C of acute myelogenous leukemia blast progenitors in a closely correlated fashion and that the cell cycle changes as well as other mechanisms are involved in the Ara-C sensitivity modulation by SCF.

Intracavity phase modulation in a fiber-optic ring laser gyro can provide 'optical dithering' to reduce the effects of frequency locking while retaining optical reciprocity in the cavity. We show that the use of two antiphased phase modulators placed symmetrically within the fiber cavity can provide uniformly distributed dithering. A modulation index of 2.4 theoretically eliminates the zero-order lock-in band around zero frequency, while use of a high modulation frequency puts higher-order lock-in bands outside the beat frequency dynamic range. Push-pull modulation allows for high modulation frequency with minimum dynamic perturbation of the cavity resonant behavior. We describe results with an experimental Brillouin fiber optic gyro operating at 1.3-micron wavelength using push-pull modulation together with a novel synthetic heterodyne detection scheme for sensing rotation rate and direction. A ten-fold reduction of the width of the zero-order lock-in band is observed. We also demonstrate that the observed frequency bias at zero rotation rate is caused by the Kerr effect due to the power imbalance between the two oppositely directed circulating lasers.

The epigenetic phenomenon could play a role in the interaction between chromatin and DNA-binding enzymes, allowing us to consider an association between the phenomenon and gene rearrangement. The correlation between methylation status and rearrangement of the T-cell receptor (TCR) beta chain gene in leukemia cells obtained from patients with acute myeloid leukemia (AML) was examined. All of the AML patients with a TCR-beta rearrangement had hypomethylated CCGG sequences within the J beta 1 region on the rearranged allele, while the germline allele had completely methylated CCmeGG sequence in this region, indicating a strong association between hypomethylation status and rearrangement of the TCR beta chain gene. In the DNA from AML patients with or without a TCR-beta rearrangement, the C beta 2 region contained completely methylated CCmeGG sequences, even though they express T-cell-associated antigens, including CD7; this pattern is quite different from that observed in T-cell neoplasias. Moreover, some AML patients showed a TCR-beta rearrangement without the presence of immunoglobulin heavy-chain gene rearrangement, suggesting that TCR beta chain gene involvement in AML is required for unknown factors other than common recombinase activity.

Epigenetic changes may play a role in genetic alterations in cancer cells, but little is known about this phenomenon. In this study we examined the correlation between rearrangement and methylation status of the T-cell receptor (TCR) beta-chain gene in 23 patients with B precursor acute lymphoblastic leukemia (ALL). In B precursor ALL, all patients had a CCmeGG sequence in the C beta 2 region, a pattern similar to that observed in normal mature B-cells. Approximately 55% of patients with B precursor ALL exhibiting a hypomethylated CCGG sequence at the J beta 1 region showed rearrangement of this region. Furthermore, the same allele of rearranged J beta 1 always contained an unmethylated sequence in the region, although another allele without rearrangement contained methylated J beta 1. By contrast, none of the patients had a rearrangement in the J beta 1 region without hypomethylation. Therefore, rearrangement of the J beta 1 region may link to the hypomethylation status of this region. A close association between hypomethylation of the J beta 1 region may promote accessibility to a putative common recombinase to produce TCR J beta 1 rearrangement. In contrast, about 45% of patients with a hypomethylated J beta 1 did not show rearrangement in this region, thus allowing categorization of B precursor ALL patients into subtypes, according to the combination of TCR beta-chain gene rearrangement and hypomethylation status, especially in the J beta 1 region.

A new human lymphoma cell line, designated DL-40, was established from the peripheral blood of a 64-year-old woman with leukemic conversion of aggressive large cell lymphoma. The cell line grew in suspension with or without forming clumps of cells and exhibited large, round, or multiple nuclei in the relatively abundant cytoplasm that was positive for acid phosphatase. The cells expressed a Ki-1 antigen (CD30), E+, CD2+, CD4+, CD45+, Ia+ phenotype and had rearranged T-cell receptor beta chain but were negative for CD15, HTLV-I, and Epstein-Barr virus nuclear antigen. Chromosome analysis of this cell line showed a human female karyotype with complex hyperdiploid abnormalities. DL-40 cells produced tumors histologically similar to the original lymphoma when transplanted into nude mice and immunosuppressed hamsters. The DL-40 cell line could provide a useful tool for the understanding of biology of the Ki-1-positive non-Hodgkin's lymphoma.

We previously reported on the heterogeneity of bone marrow stromal cell function in supporting hematopoietic cell proliferation and differentiation in vitro among refractory anemia (RA) of myelodysplastic syndrome (MDS) patients. Interestingly, stromal cells from same MDS RA patients induced an apoptotic change in CD34-positive hematopoietic cells (CD34+). To clarify the mechanism of this action of stromal cells, a cloned stromal cell line (LS801) from MDS patient was established. In a coculture system with LS801, proliferation and differentiation of CD34+ cells was inhibited. Assay for the expression of 7A6 antigen on CD34+ cells, for monitoring the cells undergoing apoptosis, showed that LS801 induced an apoptotic change in CD34+ cells. When CD34+ cells were cocultured but kept separate fom LS801 by a 0.45-mm millipore membrane to prevent their attachment, the action of LS801 for inducing apoptosis of CD34+ cells was inhibited. The expression of CD95 on CD34+ cells was induce by LS801, however, no production of fas ligand, TNF-α or IFN-γ in LS801 was observed. In light of these observations, direct contact between LS801 and CD34+ cells might be essential for maximum induction of CD34+ cells undergoing apoptosis.

"1.海藻エキス0.5%,1.5%,2.0%水溶液を水の代りにマウスに投与して発育状態を水投与の対照群と比較したところエキス投与により体重増加率が向上した.また20-メチルコランスレン処理による人工的皮膚癌を負荷し海藻エキスの発癌抑副作用を検索した結果,担癌動物において最大5週間の延命効果を認めた.2.海藻エキス中には約7 ppm, 固形飼料には0.76ppmのヒ素が含まれているが,50ppm亜ヒ酸水溶液投与群,1.5%海藻エキス投与群,水投与の対照群についてヒ素の吸収率を測定したところ海藻エキス投与群は吸収率低く,エキス投与により同時に摂取する食餌中のヒ素の吸収をも抑制することを推察した.比較のため行った亜ヒ酸水溶液投与群においてはヒ素吸収率高く飼育10週以降1/3のマウスに足部関節と筋肉の腫張を認めた.3.尿中ヒ素排泄量は50ppmヒ素水溶液投与群が最高であり,海藻エキス投与群は対照群より低く,エキス投与によって屎中へのヒ素排泄量が増加するため飼育5ヵ月現在においては生体への吸収量が少く血清中ヒ素濃度が低下し尿中への排泄量も減少するものと推察した.本実験に際し組織の病理学的鑑定を賜った三重大学医学部病理学伊豆津公作教授および実験に協力頂いた庄田節子,沢田京子様に感謝の意を表す次第である."

Erythropoietin was measured by exhypoxic polycythemic mouse method in the course of a 64-yr-old male with renal cell carcinoma associated with erythrocytosis. Serum erythropoietin fluctuated with progression of the disease. Preoperative elevated erythropoietin (0.11 U/ml, p greater than 0.05) subsided after nephrectomy and again increased with developing lung metastasis (0.1 U/ml, p greater than 0.02). Erythropoietin was markedly increased in the tumorous extracts from primary renal cell carcinoma in the kidney (0.2 U/g, p greater than 0.01) and lung metastasis (0.8 U/g, p greater than 0.01). Renal cell carcinoma from the lung metastasis was transplanted into nude mice, resulting in erythrocytosis in some of these mic. In the erythrocytotic mice, erythropoietin was elevated to levels of 0.25--0.9 U/g (p greater than 0.01) in the tumorous extracts and increased (0.67 U/ml, p greater than 0.02) in the serum. These results indicate that this renal cell carcinoma is an erythropoietin-producing tumor, and this tumor has been successfully transplanted in nude mice for the first time.

"5.8ppmのヒ素を含む海藻エキスを1.5%に希釈したものを水の代りに投与して10ヵ月飼育したマウスの成長と肉眼的症状および組織の病理学的変化を観察するとともに諸組織のヒ素含有量を測定した.また同エキスを1カ年以上連日飲用した成人の毛髪ヒ素含有量を一般成人のそれと比較した.これらの実験結果は次のとおりである. 1)1日当り海藻ヒ素7.3μgを摂取したマウスは飼育6ヵ月で9頭中1頭に足部関節の腫張を呈したが,その症状は50ppmの無機態ヒ素投与3ヵ月で出現した症状と同様であった. 2)足部の異常症状を呈したマウスは無症状のものよりヒ素吸収率,尿中ヒ素濃度,諸組織のヒ素含有量高くとくに尾部,肝組織のヒ素が腸壁,皮膚および体毛より高値であった.また赤脾髄での巨核球の増生,肝細胞におけるミトコンドリアの膨化と伸長,滑面小胞体の増加と拡張等の現象を認めた.3)連日20～30mlの海藻エキス(ヒ素含量5.8ppm)を1年以上5カ年飲用した成人の毛髪ヒ素含有量は対照区の一般成人に対し有意差を認めなかった.", "A specimen of algae extract containing 5. 8 p.p.m. arsenic was given perorally to the mice to examine its effect on the growth and any sign of intoxication of the animals. Each groups consisting of 9 mice respectively were fed commercial solid stock diet together with a sample solution of 1.5% algae extract. After 10 months feeding, the animals were killed and several organs and tissues were excised to assay their arsenic contents and also to investigate their ultrafine structure by electron microscopy. The results were as the following. 1. In the animals which had taken for 6 months daily 7. 3μg arsenic in the form of algae extract developed some swelling of their paws, the symptom of which was similar to that shown by the animals which had taken 50 p.p.m. arsenic for 3 months daily. 2. In the animals with paw swelling, the arsenic content of the tail, liver, and pawjoints were found higher than those of the intestinal wall, skin and hair. 3. The electron micrograph of the hepatic cells revealed swelling and elongation in some of the mitochondria and an increase in number of smooth microsoma and nuclei. 4. The hair from the adult persons who were given 20～30 ml sample solution of algae extract for 1～5 years daily showed on an average a similer arsenic value to that of the hair from normal adult persons as a standard"

The linkage of the plasma protein (Gl-1) and hooded loci has been determined from backcross progeny of the initial crosses Long Evans/Orl×BN/Orl and Long Evans/Orl×Fatty/Orl. The two sets of data show a significant linkage heterogeneity with, respectively, 5.0±2.8 and 15.1±3.0 recombination. This is the fourth linkage of biochemical markers so far demonstrated in the rat.

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Experimental data presented in this study indicate that prolonged pressure exerted on the kidneys of rabbits may lead to true erythrocytosis. With approximately 60 per cent of rabbits developing erythrocytosis, the pressure of experimentally induced unilateral hydronephrosis appears to be most effective in this respect. It is suggested that of the role of the renal medulla in the production of erythrocytosis is an important one. Interference with renal arteries and veins did not result in erythrocytosis. The role of erythropoietin and the possible mechanisms by which "renal pressure erythrocytosis" is produced is discussed.

1. Lactobacillus salivariusと同定された乳酸桿菌Sa-S株について,マイトマイシンCおよび,紫外線による誘発をこころみ,それが溶原株であることを確認した。2. マイトマイシンCおよび紫外線の誘発条件について検討した結果,マイトマイシンCの濃度は37C 15分処理の場合,最終濃度2∼10mcg/mlが適当であり,紫外線誘発の場合には(15W殺菌燈,90cm) 5.4erg/cm2/sec 30∼60秒の照射が有効であつた。3. 誘発の際の菌のageは対数増殖期初期が適当であり,休止期に近いものは誘発がかからなくなる。4. Sa-S株のテンペレートファージPLS-1の指示菌について検討した結果,Sa-S株の誘発生残菌より得られた非誘発性の菌株のみが指示菌となることが明らかになつた。5. PLS-1ファージの潜伏期は約80分,平均放出量は100∼130である。その他,紫外線耐性,形態,種々の乳酸桿菌に対する吸着等,PLS-1ファージの生物学的性質について検討した。

Lactobacillus casd YIT0091の多剤耐性変異株L. casei PSR3002の人工消化液およびNa-deoxycholateに対する耐性の変化を原株と比較するとともに, 他のLactobacillus, StreptococcusおよびBifidobacteriumの人工消化液耐性を調べ次の成績を得た。1. pHを変えた人工胃液に対する耐性にはL. casei YIT0091およびその耐性変異株間に差がなく, Na-deoxycholate耐性も等しかつた。しかしpH2.7～pH2.5の人工胃液で3時間処理し, 人工腸液に継代した場合, 生菌数の消長に差がみられL. casei PSR3002の誘導期が長くなり, 増殖性もやや低下した。2. 人工胃液耐性は, 供試菌中のL.caseiはすべて強く, L. acidophilusの2株, L. plantarurn YIT0062およびStreptococcus faecalis ATCC10541がこれに次いだ。3. Na-deoxycholate耐性はS. faecalis ATCC10541がもつとも強くL. casei YIT0091, L. casei PSR3002, L. casei I-136,. L. plantarum YIT0062およびL. acidophilus A-34がこれに次ぎ, その50%発育阻止濃度は330～600mcg/mlであつた。人工腸液での増殖性もL. acidophilus A-34を除き, これらの株がよかつた。S. therrnophilus YIT2001, L. bulgaricus YIT0098, L. jugurti LJ-4は人工腸液ですべて増殖せず, Nadeoxycholateの50%発育阻止濃度は55～90mcg/mlであつた。

The resting membrane of a barnacle muscle fiber is mostly permeable to cations in a solution of pH 7.7 whereas it becomes primarily permeable to anions if the pH is below 4.0. Mechanisms of ion permeation for various monovalent cations and anions were investigated at pH 7.7 and 3.9, respectively. Permeability ratios were obtained from the relationship between the membrane potential and the concentration of the test ions, and ionic conductances from current-voltage relations of the membrane. The permeability sequence for anions (SCN > I > NO3 > Br > ClO3 > Cl > BrO3 > IO3) was different from the conductance sequence for anions (Br, Cl > ClO3, NO3 > SCN). In contrast, the permeability and conductance sequences were identical for cations (K > Rb > Cs > Na > Li). The results suggest that anion permeation is governed by membrane charges while cation permeation is via some electrically neutral mechanism.

Two new esterase polymorphisms have been described in tissue homogenates of the four rat strains August/Orl, LEW/Orl, Long Evans/Orl, and WAG/Orl. Independent expression of these polymorphisms in the different strains and in various tissues of a particular animal agrees with the separate genetic control hypothesis; the gene symbols Es-4 and Es-5 have been assigned to the loci. Transmission of both genes follows a normal autosomal recessive Mendelian pattern; preliminary data indicate tight linkage between these loci and the previously described Es-2 and Es-3 loci.