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Advances in immunosuppressants enable organ transplantation for sensitized patients. However, influences of pre-formed donor-specific anti-human leukocyte antigen (HLA) antibodies (DSA) have not been fully understood in renal transplantation (RT). On the other hand, immunocomplex capture fluorescence analysis (ICFA) is a reliable method to detect donor-specific anti-HLA antibodies and HLA antigen complexes. Graft ICFA can detect DSA in an allograft (g-DSA).To elucidate the consequences of pre-formed DSA, 198 patients who underwent living-donor RT were enrolled for this study (observation period: 57.8 ± 34.9 months); 187 patients in the DSA- group (excluding ABO-incompatible cases) and 11 patients in the DSA+ group. Before RT, all DSA+ patients had undergone rituximab administration and plasmapheresis. For a graft ICFA, the biopsy specimen (1 × 10There were no significant differences in 5-year graft survival (87.9%/100% in the DSA-/DSA+ groups, respectively). In terms of antibody-mediated rejection (AMR), within 1 month after RT, pathologically determined AMR occurred 3.2% and 63.4% in the DSA- and DSA+ groups, respectively (P .0001). However, interestingly, more than half of them (57.1%) indicated only subclinical AMR, that is, no fluctuation of S-Cr. As representative of 2 cases of subclinical AMR, g-DSA deposition could be confirmed (1.15 ± 0.04) at 1 hour after reperfusion by graft ICFA. Furthermore, g-DSA shifted to 2.20 ± 0.98 at 3 weeks after transplantation, along with a decline in s-DSA mean of fluorescence intensity (1718-506.5).Although pathologically determined AMR occurred more frequently in pre-formed DSA+ recipients, it can be argued that a successful de-sensitization protocol inhibits further production of DSA and graft destruction.

The spatial distributions of potassium and rubidium in Lentinula edodes mushrooms contaminated with radioactive cesium were measured using particle-induced X-ray emission (PIXE) analysis by a beam scanning technique with a submillimeter sized proton beam to study relationships in elemental concentration in the mushrooms between radioactive cesium and other alkali elements. The results of PIXE analysis were compared with the spatial distribution of radioactive cesium in the samples measured by an autoradiographic method. Radioactive cesium was not uniformly distributed and strongly concentrated in the peripheral region of pileus of the mushroom while the elemental maps evaluated from PIXE analysis showed that potassium and rubidium are highly accumulated in the same area as radioactive cesium. The present results suggest a common mechanism of the accumulation of alkali elements including radioactive cesium in Lentinula edodes mushrooms.

A neural network has been successfully implemented in an active-mode millimeter-wave (60 GHz) imaging system with a Yagi-Uda antenna array in order to recognize objects and reconstruct images that appear distorted under coherent millimeter-wave illumination. With 10 /spl times/ 10 sampling points and five teaching trials, a recognition rate of 98% has been obtained for ten dissimilar alphabetical letters used as objects. The success rate of reconstruction of distorted millimeter-wave images was 80% when five dissimilar letters were used for the reconstruction. The recognition rate after changing the spatial resolution of the optical system and sampling interval of the image is also discussed.

A millimeter-wave optical system for plasma diagnostics for measuring the two-dimensional distribution of electron density in the Tsukuba GAMMA 10 tandem mirror has been developed. Two-dimensional images of plasma density of a cross-sectional area of 83 mm×67 mm in the machine axis direction were successfully measured with the optical system. The position at which the present imaging system is installed corresponds to the plug region, and the plasma radius is relatively narrow (less than 100 mm). Information about the density in the area is valuable in the study of confining-potential formation. This optical system was developed for a millimeter-wave phase-imaging interferometer and was designed with the ray tracing method to reduce aberration. It was evaluated experimentally by measuring images of point sources and phase images of dielectric targets. The receiving optical system from the plasma to detector array was designed with a spatial resolution of 28.7 mm and a magnification of 0.30. Ray tracing in...

Millimeter-wave imaging systems in the frequency range of 70–140 GHz have been developed for diagnostics of magnetically confined plasmas. The 70 GHz imaging system successfully measures time evolutions of both radial and axial profiles of line density and electron cyclotron emission (ECE) in a tandem mirror. The imaging system is being installed in Large Helical Device (LHD) at the National Institute for Fusion Science. In order to cover the frequency range of the second harmonic ECE on LHD, a new detector using monolithic microwave integrated circuit technology has been designed and fabricated. The detector consists of the integration of a receiving antenna, a down-converting mixer diode, and an intermediate frequency amplifier on a GaAs substrate chip. The heterodyne response up to 10 GHz was confirmed at 70–140 GHz. The optical system consisting of an ellipsoidal mirror and a flat mirror was designed by using a ray-tracing code and evaluated experimentally at 140 GHz.

The induced proliferative response of the human derived neuroblastoma cell (IMR32), with the cell proliferation medi- ating factor, is much milder compared to other cell strains, and develops cellular clusters which are characterized by numerous extensions of dendrites. Presuming this phenomenon to be one of the induced neuronal differentiation due to genetic alterations , we preliminary studied to elucidate the changes in gene expression of the IMR32 by mRNA differential display analysis, moni- toring polymerase chain reaction (PCR) products by non-gel molecular sieving capillary electrophoresis in linear polyacrylamide solution. The CE fingerprints revealed a number of peaks with differential expression patterns.

A millimeter-wave two-dimensional imaging array has been developed for diagnostics of magnetically confined plasmas. It is applied to the plug cell of the GAMMA 10 tandem mirror, and measures time evolution of radial and axial line-density and electron cyclotron emission (ECE) profiles in one plasma shot. The monolithic-type GaAs Schottky diode detector having an antenna and built-in amplifier on a GaAs substrate is newly fabricated using monolithic microwave integrated circuit (MMIC) technology. This novel detector is developed to attain high frequency response up to submillimeter-wave region and wideband heterodyne characteristics, and is expected to be applied to ECE imaging in a large helical device. The response of the detector is compared with that of the hybrid-type detector using beam-lead Schottky barrier diodes. The first experimental result of the ECE measurement is reported using a heterodyne radiometer with the MMIC detector.

Pyrolytic carbon was infiltrated into porous carbon or two-dimensionally woven carbon fibre (2D-C)/SiC particulate preforms using pressure-pulsed chemical vapour infiltration from C6H6 (3–16%)–H2–N2 at 1273–1373 K. Residual porosity of porous carbon decreased from 29 to 10% after 1×104 pulses at 1323 K, and that of 2D-C/SiC particulate preform decreased from 30 to 7.5% after 4×104 pulses at 1273 K. Flexural strength of 2D-C/SiC preform reached about 150 MPa.

A novel lateral power device, termed a p-channel dual-action device (p-ch DAD), is proposed and experimentally demonstrated in action. This device is based on a new dual-action mechanism. The new device has successfully increased on-state current without lowering the device breakdown voltage. The 600-V level-shifting action of the p-ch DAD has been confirmed by a circuit experiment. A newly designed p-ch DAD on the silicon on insulator can be made by adding four additional masks and trench technology to a 0.8-/spl mu/m CMOS process. Moreover, the process we have developed is completely compatible with an existing 5-V 0.8-/spl mu/m CMOS process.

This paper proposes a visual navigation method for humanoid robot, based on the probabilistic multiple stereo matching. In this paper, we assumed that the existence of measured obstacle is probabilistic. We also consider that stereo vision has high reliability range for location estimation according to the distance between cameras. We therefore propose a stochastic model to measure obstacle location using successive parallactic images in different distance between cameras. We then implement a navigation system for humanoid robot, HOAP-1, using proposed model. This paper also reports performance results of autonomous walk to reach goal position avoiding several obstacles.

A novel robot, Ifbot, which can communicate with human by joyful conversation and emotional facial expression, has been developed by our industry-university joint research project. In this paper, as the fundamental technology of Ifbot for the robot-human interaction, we propose an image processing method for real-time face tracking and talker distinction.

F10, a subline of the B16 mouse melanoma cell line, is itself the parent of the more metastatic BL6 line. BL6 cells differ from F10 cells by an alteration of the gene encoding the B56gamma regulatory subunit of protein phosphatase 2A (PP2A), which results in mRNA encoding a truncated variant of the subunit (deltagamma1). Expression of deltagamma 1 protein is detectable only when BL6 cells are transplanted into mice and then gamma-irradiated. Recently, B56gamma subunit-containing PP2A holoenzymes have shown to dephosphorylate Mdm2, a negative regulator of p53. Thus, we assessed whether the expression of deltagamma1 affects irradiation-induced phosphorylation of Mdm2 and radioresistance of melanoma cells by perturbing the regulation of p53. Western blot analyses revealed that irradiated COS-7 and NIH3T3 cells stably expressing deltagamma1 showed significantly less irradiation-induced Mdm2 phosphorylation. Mdm2 phosphorylation reduces the ability of Mdm2 to target p53 for degradation, which probably explained why p53 protein levels in NIH3T3 cells expressing deltagamma1 were not significantly elevated by irradiation, unlike in wild-type cells. This was also true for F10 cells transfected with deltagamma1 (F10deltagamma1) when the cells expressed deltagamma1 after being irradiated in vivo. p53 mRNA levels in irradiated wild-type and deltagamma 1-expressing cells were both only slightly elevated, suggesting that Mdm2 regulates p53 levels by a post-transcriptional mechanism. p53-mediated induction of the pro-apoptotic gene encoding Bax was also significantly lower in F10deltagamma1 cells irradiated in vivo. Moreover, F10deltagamma1 and BL6 cells were less apoptotic than F10 cells when the cells were irradiated in vivo. The p53 in F10 cells appears to be as functional as that in NIH3T3 cells because irradiation-induced expression of p53-target genes was comparable in both cells. Collectively, deltagamma1 appears to reduce irradiation-induced Mdm2 phosphorylation, which then blocks irradiation-stimulated p53 accumulation. Defects, such as deltagamma1, in PP2A may thus contribute to melanoma cell radioresistance.

F10, a subline of the B16 mouse melanoma cell line, is itself the parent of the more metastatic BL6 line. BL6 cells differ from F10 cells by an alteration of the gene encoding the B56g regulatory subunit of protein phosphatase 2A (PP2A), which results in mRNA encoding a truncated variant of the subunit (Dg1). Expression of Dg1 protein is detectable only when BL6 cells are transplanted into mice and then g-irradiated. Recently, B56g subunit-containing PP2A holoenzymes have shown to dephosphorylate Mdm2, a negative regulator of p53. Thus, we assessed whether the expression of Dg1 affects irradiation-induced phosphorylation of Mdm2 and radioresistance of melanoma cells by perturbing the regulation of p53. Western blot analyses revealed that irradiated COS-7 and NIH3T3 cells stably expressing Dg1 showed significantly less irradiation-induced Mdm2 phosphorylation. Mdm2 phosphorylation reduces the ability of Mdm2 to target p53 for degradation, which probably explained why p53 protein levels in NIH3T3 cells expressing Dg1 were not significantly elevated by irradiation, unlike in wild-type cells. This was also true for F10 cells transfected with Dg1 (F10Dg1) when the cells expressed Dg1 after being irradiated in vivo. p53 mRNA levels in irradiated wild-type and Dg1-expressing cells were both only slightly elevated, suggesting that Mdm2 regulates p53 levels by a post-transcriptional mechanism. p53-mediated induction of the pro-apoptotic gene encoding Bax was also significantly lower in F10Dg1 cells irradiated in vivo. Moreover, F10Dg1 and BL6 cells were less apoptotic than F10 cells when the cells were irradiated in vivo. The p53 in F10 cells appears to be as functional as that in NIH3T3 cells because irradiation-induced expression of p53-target genes was comparable in both cells. Collectively, Dg1 appears to reduce irradiation-induced Mdm2 phosphorylation, which then blocks irradiation-stimulated p53 accumulation. Defects, such as Dg1, in PP2A may thus contribute to melanoma cell radioresistance.

Studies have suggested that 17beta estradiol (E2) can modify apolipoprotein E (apoE) expression. The current study determined if apoE protein varied in different regions of the mouse brain as a function of the estrous cycle and if E2 could increase apoE protein expression. In this study apoE concentration was lowest on estrus in the hippocampus, cingulate cortex and frontal cortex. In contrast, apoE concentration was highest on estrus in the olfactory bulb and cerebellum. There were no differences in the striatal apoE expression throughout the estrous cycle. Exogenous E2 significantly raised tissue levels of apoE in the olfactory bulb and cerebellum at 5 days after treatment. There was a slight, but nonsignificant increase in cortical expression of apoE and no change in striatum. Immunocytochemical localization studies found estrogen receptor alpha (ERalpha) and estrogen receptor beta (ERbeta) in cortical neurons and glia. In the cerebellum and olfactory bulb, ERbeta was seen primarily in glia. ERalpha was not observed in the cerebellum and was rare in the olfactory bulb. Neither ERalpha nor ERbeta was seen in the striatum. Our data show regional differences in the production of apoE throughout the estrous cycle. In addition, exogenous E2 has regionally specific effects on apoE expression. Regional variability in apoE production appears to vary as a function of the estrogen receptor subtype.

The BL6 subline was derived from the F10 line, which was derived from the B16 mouse melanoma cell line. BL6 cells are more invasive than F10 cells and differ genetically from F10 cells by an alteration of the gene encoding the B56gamma regulatory subunit of protein phosphatase 2A (PP2A). This alteration results in the transcription of mRNA encoding a truncated variant of the B56gamma1 isoform (Delta-gamma-1). Delta-gamma-1 is capable of targeting PP2A to the specific subcellular sites but incapable of promoting the dephosphorylation of specific substrates that is normally mediated by the B56gamma subunit-containing PP2A holoenzyme. It thus appears that activities of this type of holoenzymes decrease in cells expressing Delta-gamma-1. Recently, we found two possible ways how Delta-gamma-1 contributes to the enhanced metastatic potential of BL6 cells. The two ways seemed far away from each other: Delta-gamma-1 influenced both the nuclear and cytoplasmic functions of the cell. In the cytoplasm, Delta-gamma-1 localized at the Golgi complex and accelerated Golgi-mediated vesicle transport. On the other hand, Delta-gamma-1 disturbed the cell-cycle regulation. In response to gamma-irradiation, protein levels of Delta-gamma-1 were markedly increased in BL6 cells. Subsequently the integrity of cell-cycle checkpoint became more aberrant in BL6 cells than that in F10 cells. These two actions of Delta-gamma-1 could results in the enhancement of the malignant phenotypes of melanoma cells, as discussed in this review.

Members of the immunoglobulin superfamily often play key roles in intercellular adhesion. IGSF4 is a novel immunoglobulin (Ig)-like intercellular adhesion molecule. Three Ig-like domains are included in the extracellular domain of IGSF4 and mediate homophilic or heterophilic interactions independently of Ca2+. The cytoplasmic domain of IGSF4 contains the binding motifs that connect to actin fibers. Since IGSF4 has been characterized by several independent research groups, this molecule is called by three names, TSLC1, SgIGSF and SynCAM. IGSF4 was first characterized as a tumor suppressor of non-small cell lung cancer and termed TSLC1, although how IGSF4 suppresses tumor growth remains unknown. Silencing of the IGSF4 gene was primarily achieved by allelic loss and promoter methylation in this type of cancers. Soon after this discovery, IGSF4 was found to have roles in adhesion of spermatogenic cells to Sertoli cells and mast cells to fibroblasts and termed SgIGSF. Other researchers revealed that IGSF4 drives synaptic formation of neural cells and termed it SynCAM.

We have demonstrated that the developed process has a breakdown voltage of higher than 600 V with use of thick buried-oxide and thin SOI. From both experiments and simulations, the cylindrical structure in the LIGBTs shows the best performance; it improves the latch-up tolerance without the increase of on-state voltage. Moreover, the process we have developed is completely compatible with an existing 5 V, 0.8 /spl mu/m CMOS process.

To test the stability of the electromotive force (EMF) of the solid electrolyte CO/sub 2/ sensor with Na/sub 2/CO/sub 3///NASICON structure, the potential variations at the cathode (the NASICON/gas interface) and anode (the gas/Na/sub 2/CO/sub 3/ interface) of the cell were separately measured by setting a reference electrode. In air, the EMF of the sensor showed a significant deviation, which was caused by the instability of the cathode electrode. Electrode probe micro analysis (EPMA) detected the formation of Na/sub 2/SO/sub 4/ at the cathode. A novel sensor with a structure in which the NASICON was covered with yttria-stabilized zirconia (YSZ) at the cathode was proposed to prevent the formation of Na/sub 2/SO/sub 4/. This sensor exhibited excellent long-term stability and works well, according to the Nernst equation, in air. >

A novel lateral power device termed the p-channel dual action device (p-ch DAD) is proposed and demonstrated in action experimentally and numerically. It consists of a p-channel HVMOS, n-channel HVMOS and resistance. This hybrid device has increased on-state current without lowering the device breakdown voltage. The conductivity modulation of the new device is discussed.

A novel lateral power device termed p-channel Dual Action Device (p-ch DAD) is proposed and demonstrated in action experimentally. The new device has successfully increased on-state current without lowering the device breakdown voltage. 600 V level-shifting action of the p-ch DAD has been confirmed by a circuit experiment.

A new p-channel slit type p-drift lateral IGBT (p-ch SD-LIGBT) is described. This device is fabricated with thick buried-oxide SOI, using 0.8 /spl mu/m CMOS process. The improvement of slit type p-drift structure makes it possible to realize excellent characteristics in the trade-off between breakdown voltage and on-state current relationship compared with p-ch conventional SOI-LIGBT. Furthermore, measured characteristics are reported for p-ch SOI-LIGBT with a breakdown voltage of more than 500 V for the first time.

A newly designed diode is proposed and demonstrated in action experimentally. The new device has successfully increased forward current without additional mask and increase of the area. Remarkable improvement in the reverse recovery characteristic of the new diode can be realized by electron irradiation. The fabricated ICs with the new diode operated at 3 kHz PWM carrier frequency.

This paper reports on the recent results of two types of millimeter-wave plasma diagnostics, reflectometry and imaging, applied to a tandem mirror plasma. The ultrafast-sweep broadband and ultrashort-pulse reflectometry provide reliable density profiles under the existence of density fluctuations. The combination of high performance system and advanced data analysis methods are necessary for automatic evaluation of density profiles. A millimeter-wave imaging array in the frequency range of 60-90 GHz has been successfully applied for measurements of line-density and electron cyclotron emission (ECE) profiles. It is being installed in Large Helical Device (LHD). In order to cover the frequency range of the second harmonic ECE on LHD (120-180 GHz), a new detector using MMIC technology has been designed and fabricated.

Cancer develops and progresses as genetic alterations occur subsequently. Onset process of cancer has become well understood in some types of cancer, such as colorectal cancers. In this process, responsible alterations were identified in numbers of oncogenes such as k-ras, and tumor suppressor genes such as p53, as Vogelstein proposed earlier in the multistage carcinogenesis theory. In contrast, our understanding remains short to draw such an adequate diagram for the process during which cancer becomes more malignant, i.e., metastatic. To examine the molecular basis for this progression step, mouse metastasis models have been established where tumor cell lines are inoculated into mice and metastasize to specific organs. The model using B16 melanoma cells is one of the most developed. BL6 subline, one of the most metastatic, was obtained from F10 subline simply through six rounds of in vitro selection. Nonetheless, BL6 cells metastasize lungs much more heavily than F10 cells when injected subcutaneously. The difference in gene expression between the two sublines is considered rather small but relevant for spontaneous metastasis. We began our research by elaborating a method for the construction of subtracted cDNA libraries, and made it applicable to BL6 and F10 cells. As a result, we were able to isolate a couple of genes that were expressed differently between the two sublines. As might be expected, each of the genes appeared to play a role more or less in distinct aspects of spontaneous metastasis of B16 melanoma cells. Moreover, similar roles were expected for the genes in the process by which human melanoma cells metastasize.

The sporulation program in Bacillus subtilis ends in the formation of a highly resistant endospore that can withstand extremes of heat, mechanical disruption, ultraviolet irradiation, lytic enzymes and chemical attack. These properties are attributed mainly to the unique structure of spore coat and cortex, as well as to the physical state of the spore cytoplasm. The outermost layer of the spore, called the coat, has two morphologically distinct sublayers: an electron-dense outer coat and an electron-translucent inner coat. The coat is composed of more than 2 dozen proteins of varying size. Many coat genes and coat proteins have been isolated and characterized in detail, and studies of these have identified proteins with important roles in coat assembly, resistance and spore germination. We describe here characteristics of the coat proteins and propose a model for coat assembly based on recent work.

F10 and BL6 cells of B16 mouse melanoma cells are metastatic after intravenous injection, but only BL6 cells can metastasize to lungs after subcutaneous injection. Differences in gene expression between the two cell lines were examined, and a greater expression of the Sik-similar protein (Sik-SP) gene was found in BL6 cells. Structurally, Sik-SP belongs to the nucleolar Nop5/Sik family whose members play central roles in ribosome biogenesis; however, the function of Sik-SP has not been examined. Cytology with green fluorescent protein-fused proteins showed that Sik-SP was localized to the nucleolus. To examine whether Sik-SP is involved in ribosome biogenesis, two parameters were measured: magnitude of ribosomal RNA synthesis per nucleus and magnitude of protein production from the same amount of mRNA of an exogenous luciferase gene. Both values and, in addition, nucleolar size were larger in COS-7 monkey kidney cells overexpressing Sik-SP and BL6 cells than in mock-transfected COS-7 and F10 cells, respectively. Sik-SP seemed to promote ribosome biogenesis in the nucleolus. Furthermore, the expression of Sik-SP seemed to confer a greater cell growth response to serum, because such a response was greater in BL6 cells and F10 cells overexpressing Sik-SP than in untreated and mock-transfected F10 cells. Sik-SP may render melanoma cells more competent to survive through augmenting the activity of nucleolus.

Ciliary neurotrophic factor (CNTF) can prevent injury-induced motor neuron death. However, it is also evident that expression of CNTF in Schwann cells is suppressed during nerve regeneration. In this report, we have addressed the mechanism underlying the down-regulation of CNTF expression in injured nerves using a mouse Schwann cell line IMS32 and mouse sciatic nerve. In IMS32 cells, activation of the Ras extracellular-signal-regulated kinase (ERK) pathway by adenoviral vector-mediated expression of dominant active MEK1 did not alter a basal level of CNTF expression, whereas inhibition of the Ras-ERK pathway by using adenoviral vectors resulted in a marked increase in CNTF expression. This inverse relation between before and after axotomy was also observed in mouse sciatic nerve. In the axotomized sciatic nerve, the phosphorylated ERK was markedly increased; in contrast, the expression of CNTF was markedly decreased. These findings suggest that an inactive state of ERK is crucial for the CNTF expression in Schwann cells, and that activation of ERK following nerve injury critically influences the expression of CNTF. This might well explain why CNTF is highly expressed in quiescent Schwann cells in the peripheral nervous system, and also why CNTF is not abundant in axotomized nerves or cultured Schwann cells in which the proliferation signal is obviously active.

Much of the lethality of malignant neoplasms is attributable directly to their ability to develop secondary growths in organs at a distance from the primary tumor mass, whereas few patients die from their primary neoplasm. Little is known about the molecular mechanism of tumor metastasis, however, which is controlled by a variety of positive and negative factors. In the search for metastasis suppressor genes, we have used the microcell-mediated chromosome transfer method and a rat prostate tumor model in SCID mice. When human chromosome 2 was introduced into the highly metastatic rat prostatic tumor cell, AT6.1, the metastatic ability of this cell was significantly (99%) decreased in animals. An STS-based PCR analysis for 8 hybrid clones indicates that the suppressor activity is located in the p25-22 region of the chromosome. Furthermore, the AT6.1 cell with human chromosome 2 showed a reduced ability to invade Matrigel, suggesting that the suppressor activity is involved in the step of tumor invasion during the progression of prostate cancer. We have also examined the status of the suppressor region on chromosome 2 in human prostate cancer specimens and found that this region was often lost in high-grade tumors. These results suggest that the putative suppressor gene on chromosome 2 is functionally involved in the progression of human prostate cancer. Genes Chromosomes Cancer 28:285-293, 2000.

Genomic aberrations at the chromosome 16q arm are one of the most consistent abnormalities observed by loss of heterozygosity and comparative genomic hybridization analyses in human prostate cancer, suggesting that there are tumor suppressor or metastasis suppressor genes encoded by this chromosomal region. To functionally identify such suppressor genes, we have conducted microcell-mediated chromosome transfer to introduce human chromosome 16 into the highly metastatic Dunning rat prostatic cancer cell line, AT6.1. The metastatic ability of the resultant microcell hybrid clones was then tested in a standard spontaneous metastasis assay using SCID mice. When the microcell-mediated chromosome transfer hybrid cells containing whole human chromosome 16 were injected, the number of metastatic lesions in the lung was significantly reduced as much as 99% on average. Therefore, chromosome 16 has a strong activity to suppress the metastatic ability of AT6.1 cells while it did not affect the tumorigenesis and tumor growth rate. A PCR analysis of various microcell hybrid clones with sequence-tagged site markers indicates that the metastasis suppressor activity is located in the q24.2 region of chromosome 16. Our results are consistent with the previous finding that the region of human chromosome 16q has frequent loss of heterozygosity in prostate cancer patients and suggest that there is a metastasis suppressor gene in this region that may play an important role in the progression of prostate cancer.

In Hokkaido, Japan, in order to investigate the etiological relation between gastric cancer and lifestyle, a case-control study was conducted, employing 242 cases with this cancer and 484 controls matched for sex, age, and place of residence. Eating meals within a short time, eating until full, irregularity in the time for supper, having dentures, and lack of teeth showed high odds ratios. Putting an excess digestive burden on the stomach by insufficient chewing promotes the development of this cancer. Stress, long working time, irregularity in sleeping time, and gray hair in males showed high odds ratios. A stressful life may be promotive for this cancer through depressed immunoresistance. Logistic regression analysis showed that those who ate meals rapidly, had strong stress and had lost many teeth were at high risk. The linear trend of the dose-response relationship between the number of teeth lost and the odds ratio was highly significant. To avoid stress, to chew foods sufficiently, and to maintain the health of the oral cavity can help to prevent gastric cancer.

There is a critical need for markers that can be used to predict accurately the malignant potential of histological prostate cancers (J. T. Isaacs. Am. J. Pathol., 150: 1511-1521, 1997). Metastasis-suppressor genes are attractive candidates for marker development because, by definition, their loss should be associated with the acquisition of metastatic ability. In an effort to identify such genes, a single copy of human chromosome 12, tagged with the neomycin resistance gene, was introduced into highly metastatic Dunning AT6.1 prostate cancer cells by microcell-mediated chromosomal transfer. Thirty-two AT6.1-12 clonal cell lines were established and the region(s) of chromosome 12 retained was determined by sequence tagged site-based PCR analysis. Representative AT6.1-12 clones containing overlapping regions of chromosome 12 were characterized cytogenetically and were shown to have a normal complement of parental AT6.1 rat chromosomes. Fluorescence in situ hybridization, performed on representative AT6.1-12 hybrids, demonstrated a single human chromosome 12-specific signal. The metastatic ability of six representative clones was tested in immunodeficient mice. All of the AT6.1-12 clones showed the same in vivo growth rates as the control AT6.1-neo cells. Clonal cell lines that contained a conserved approximately 70-cM portion of chromosome 12 (e.g., AT6.1-12-8, -8-1, and -8-3), showed a30-fold suppression in the number of macroscopic surface lung metastases. Mice that received injections of these cells developed a mean number 4 lung metastases whereas mice that received injections of other AT6.1-12 hybrids (lacking the approximately 70-cM region) or AT6.1-neo control cells, developed a mean number of 140 metastases. Interestingly, histological examination of the lungs of the mice that received injections of AT6.1-12-8 cells showed essentially no microscopic metastases. These findings suggest that a gene(s) encoded by the approximately 70-cM portion of human chromosome 12 suppresses an early step in the metastatic cascade.

Prostate cancers account for 43% of all cancers diagnosed in American men. It is estimated that in 1996, 317,000 new cases of prostate cancer were diagnosed and 41,000 men died of the disease. The challenge of treating prostate cancer lies in accurately distinguishing those histologically-localized cancers which will complete metastatic progression from those that will remain indolent. At this time, we lack appropriate histological markers to make such distinctions, therefore, it is often difficult to accurately predict the clinical course of an individual patient's disease. There is growing evidence that a critical event in the progression of a tumor cell from a non-metastatic to metastatic phenotype is the loss of function of metastasis-suppressor genes. These genes specifically suppress the ability of a cell to metastasize. Work from several groups has demonstrated that human chromosomes 8, 10, 11 and 17 encode prostate cancer metastasis suppressor activities. As a result of these efforts the first prostate cancer metastasis-suppressor gene, KAI1, was identified and mapped to the p11-2 region of chromosome 11. In subsequent studies, an additional gene encoded by the same region, CD44 was also determined to have metastasis-suppressor activity. Recent studies have shown a correlation between decreased expression of KAI1 and CD44 and an increased malignant potential of prostate cancers. It is anticipated that the identification of other metastasis suppressor genes may allow for the development of diagnostic markers useful in the clinical substaging of individual tumors. This manuscript is intended to present our perspective on the importance of these genes in the understanding of prostate cancer progression. More importantly, we present new findings from our laboratory's effort to identify the metastasis-suppressor genes encoded by human chromosome 17. Specifically we report the strategy currently being used to evaluate a series of candidate genes and the approach being utilized to pinpoint the metastasis-suppressor region on human chromosome 17.

In order to estimate the distribution of Salmonella including Salmonella enteritidis (SE) and SE-antibodies in commercial layer hen flocks in Hokkaido, the northern prefecture of Japan, a survey of spent layer hens was performed, from August 1996 to January 1997. From the three spent hen processing plants, samples of intestines and sera were collected from 740 birds presented for slaughter from 37 flocks of 22 layer hen farms. Intestines from each birds were cultured for Salmonella including Salmonella enteritidis. Serum from each bird was examined for SE-antibody with the enzyme-linked immunosorbent assay (ELISA). Salmonella (any serotype) and Salmonella enteritidis were isolated from 50 (6.8%) and three (0.4%) of 740 birds, respectively, and SE-antibody positive values were recorded from seven birds (0.9%). SE-antibody positive birds did not always indicate isolation of Salmonella enteritidis, however SE-antibody positive hens were demonstrated only from Salmonella enteritidis positive flocks. Salmonellae were isolated from the birds of ten layer hen farms, all of these hens were raised in houses without windows and with automatic feeders. No isolations of salmonella were made from birds raised in houses with windows. From the windowless houses, Salmonellae were isolated from 46 (21.8%) of 260 birds in houses with four to six cages piled up vertically, and from six (2.5%) of 240 samples from the houses with four to five cages piled in a slanting manner.

n-alkylpolyoxyethylene surfactants (CnH2n+1O(CH2CH2O)mH; CnEm) showed a strong enhancing effect on the inactivation of lipopolysaccharide (LPS) by heat treatment over a wide range of temperatures. The effect of CnE8 (n = 10-16) was observed above the critical micelle concentration (CMC) and above the cloud point, and was influenced by the length of the alkyl chains. The efficacy of the surfactants was in the order C10E8C12E8, C16E8C14E8. However, the hydrophilic moiety seemed to have no influence. An 80-95% solution of n-butanol showed a similar effect, indicating that LPS was more effectively inactivated in the oily phase of the surfactants than in water. The effect of surfactant on the hydrodynamic diameter of LPS was the same before and after steam-heat treatment for 20 minutes at 121 degrees C. Each surfactant disaggregated LPS without alteration of the activity of LPS before the heat treatment. We consider that the surfactants interact with LPS in the region of lipid A in a manner that favors loss of the activities of LPS during heating.

1. The growth inhibitory factor (GIF) is a 68-amino acid protein which is capable of inhibiting the growth of neuronal cells in vitro. 2. We have cloned and sequenced the 5'-flanking region of the mouse GIF gene, which spans from the transcriptional initiation site to the -1854 nucleotide. 3. This region contains sequences homologous to hgcs, SPE, and the JCV silencer domain that functions in a glial cell specific manner. This region also contains two metal responding elements and putative binding sites for AP-1, AP-2, Sp-1, SP-2, NF-1, and CREB. 4. An analysis of the reporter plasmids containing the various regions of the 5'-flanking sequence revealed that the region indeed functioned in a tissue-specific manner in glial cells and that the region between -328 and 175 is responsible for suppression, while the region between -175 and -49 is involved in the activation of gene expression.

We have chemically synthesized tracheal antimicrobial peptide (TAP) with 38 amino acids and examined efficacy of the peptide on various organisms. The synthetic peptide showed potent bactericidal effect on both gram positive and negative bacteria. The action of bactericidal effect was relatively quick and 99.9% of E. coli cells were killed within 90 minutes at a concentration of 2.5 micrograms/ml of TAP. The peptide also showed antifungal activity against both mycelia (Aspergillus fumigatus) and yeast (Candida albicans) forms of fungi. Our domain analysis with a series of synthetic peptides of various lengths indicates that 17 amino acid residues of the C-terminal end is the minimum functional domain of the bactericidal activity.

Polyoxyethylene (20) sorbitan mono-fatty acid esters strongly enhanced the inactivation of lipopolysaccharide (LPS) by steam-heat treatment at 121 degrees C, as assayed by using the Limulus amebocyte lysate (LAL) and the pyrogen test. In an aqueous solution containing 0.1% surfactant, the decrease of LPS (1 microgram/ml) from E. coli 055:B5 at 121 degrees C followed first-order kinetics. Based on the LAL assay, 0.1% surfactant was essential to achieve 3-log cycle reduction of LPS with concomitant loss of pyrogenicity by steam-heat treatment for 20 min at 121 degrees C. Steam-heat treatment for 20 min at 121 degrees C in the absence of surfactant was insufficient to achieve depyrogenation. Polyoxyethylene (9) lauryl ether and decaglycerin mono-laurate similarly enhanced depyrogenation by steam-heat treatment. The effects of all the surfactants were concentration-dependent for all of the six kinds of LPS examined.

The inactivation of endotoxin from six species of smooth gram-negative bacteria (S-form) by steam-heat treatment was investigated using the Limulus amebocyte lysate (LAL) assay. Biphasic decreases of endotoxins from four species of bacteria were observed upon steam-heat treatment of 1 microgram/ml endotoxin solution at 121 degrees C in a steam sterilizer. A lag time, however, was observed in the inactivation profiles of V. cholerae and P. aeruginosa. Distinct differences in heat resistance were observed among the bacterial species. The decrease rate was found to be concentration-dependent, and endotoxins at low concentrations (less than 10 ng/ml) were inactivated by the treatment to below the detection limit of the LAL assay. The time-course of the decrease of endotoxin from rough strains (R-form) resembled that of the respective S-form. The inactivation of R-form, especially Rc mutant, endotoxin was markedly affected by divalent cations such as Mg2+ and Ca2+, which appear to promote reaggregation of the endotoxin.

Metachromatic leukodystrophy (MLD) is an inherited metabolic disease which is characterized by deficient activity of arylsulfatase A (ASA). This deficiency causes progressive accumulation of cerebroside sulfate (CS) in oligodendrocytes (OLs) in the brain resulting in dysmyelination. We are developing approaches for treating MLD based on direct delivery of ASA genes to brain. In this paper, we report a recombinant adenovirus (Adex1SRLacZL) that efficiently transduces OLs. Moreover, transduction and expression of the human transgene was efficient in primary fibroblasts from MLD patients exposed to similar recombinant adenovirus carrying the ASA gene (Adex1SRASA) In addition, the expressed ASA protein was correctly targeted to lysosomes and was immunoreactive to anti-ASA antibody. The enzymatic activity of ASA in MLD patient cells is corrected from the deficient state to 2.3 to 5.0 times the activity of normal control cells. These results demonstrate the correction of ASA deficiency by a recombinant adenovirus that is potentially useful for transferring the gene to brain.

The Hepatocyte growth factor is the most potent mitogen for hepatocytes in primary culture and is involved in liver regeneration. The expression of the gene appears to be tightly controlled by various humoral factors. To understand the molecular mechanism of the gene expression, we cloned and determined the nucleotide sequence of the 5′-flanking region of the gene. In this region, there are sequences homologous to responding elements of P53, Rb, IL-1, IL-6, glucocorticoids, TPA and TGF-β. We also identified three major transcriptional initiation sites by primer extension analysis of this region. Functional analyses of this region by constructing CAT reporter plasmids indicate that the sequence functions in a tissue specific manner and there is a negative regulatory region which suppresses the gene expression in rat transformed kidney cells.

Twenty-four germinomas from which tumor sites were 11 pineal, 7 suprasellar, and 6 basal ganglial and thalamic (BG-T) were subjected. Humoral tumor markers of HCG, CEA and AFP were measured and compared with immunohistochemistries for these tumor markers along with PLAP, HPL and PKK1. Humoral tumor markers and immunohistochemistries did not always comprehend to each other. Difference of the survival rates among tumor sites was thought to be attributed to the variabilities in their histological, immunochemical and flow cytometric results.