The distributions of five NMDA receptor channel subunit mRNAs in the mouse forebrain at postnatal... more The distributions of five NMDA receptor channel subunit mRNAs in the mouse forebrain at postnatal day 21 were semiquantitatively examined by in situ hybridization with subunitspecific oligonucleotide probes. In contrast to ubiquitous distribution of the (1 subunit mRNA throughout the forebrain, distributions of four E subunit mRNAs were highly variable from nucleus to nucleus. The telencephalon (except for the septum) expressed the €1 and €2 subunit mRNAs. Various combinations of the €1, €2, €3, and €4 subunit mRNAs were present in different nuclei of the septum, the olfactory bulb, and the thalamus. In the hypothalamus, the suprachiasmatic nucleus expressed distinct signals for the €3 subunit mRNA alone, whereas other nuclei showed faint signals for the el, €2, and €4 subunit mRNAs. Moreover, different signal levels of the E subunit mRNAs were found in various regions. The hippocampal CA1 region expressed higher signals for the €1 and €2 subunit mRNAs than the CA3 region and the dentate gyrus. In the cerebral cortex, signal levels of the €1 subunit mRNA were higher in the laminae 111111, IV, and VI than the lamina V, whereas those of the €2 subunit mRNA were highest in laminae IIiIII and lowest in laminae IV and V. Different signal levels between the €1 and € 2 subunit mRNAs were also discerned in the amygdala, the caudate-putamen, and the thalamus. The distinct anatomical distributions and differential signal levels of the E subunit mRNAs strongly suggest different subunit organizations of the NMDA receptor channel in different forebrain neurons, which may result in functional diversity of the channel in vivo.
ABSTRACT Hierarchical State Transition Matrix (HSTM) is a table-based modeling language for devel... more ABSTRACT Hierarchical State Transition Matrix (HSTM) is a table-based modeling language for developing designs of software systems. Although widely used and adopted by (particularly Japanese) software industry, there is still lack of mechanized formal verification supports for conducting rigorous and automatic analysis to improve reliability of HSTM designs. In this paper, we first present a formalization of HSTM designs as state transition systems. Consequentially, based on this formalization, we propose a symbolic encoding approach, through which correctness of a HSTM design with respect to LTL properties could be represented as Bounded Model Checking (BMC) problems that could be determined by Satisfiability Modulo Theories (SMT) solving. We have implemented our encoding approach in a tool called Garakabu2 with the state-of-the-art SMT solver CVC3 as its back-ended solver. Furthermore, in our preliminary experiments, a conceptually simple but steadily effective way of accelerating SMT solving for HSTM designs is investigated and reported.
Gan to Kagaku Ryoho Cancer Chemotherapy, Nov 1, 2012
Patient 1 was a 63-year-old woman whose chief complaint was a mass in the left breast. Physical e... more Patient 1 was a 63-year-old woman whose chief complaint was a mass in the left breast. Physical examination revealed an inverted left nipple, a very large mass on the anterior aspect of the sternum, and erythema. Because the tumor had directly invaded the sternum, T4cN3M0, stage IIIC breast cancer was diagnosed. The patient preoperatively received chemotherapy with 6 courses of FEC100 (5-fluorouracil, epirubicin, and cyclophosphamide) and 5 courses of nanoparticle albumin -bound paclitaxel (260 mg/m2), which enabled a partial response. Patient 2 was an 83-year-old woman whose chief complaint was a mass in the upper internal and external quadrants of the right breast measuring 20×15 cm and erythema. The mass was accompanied by enlarged right axillary lymph nodes(T4bN1M0, stage IIIB breast cancer). Both patients underwent core needle biopsy of the skin and breast masses. They were both diagnosed with invasive, lobular, triple-negative breast cancer (estrogen receptor negative, progesterone receptor negative, human epidermal growth factor receptor 2 negative). The surgical resection line was drawn to include the extensive skin invasion, and mastectomy and axillary dissection were performed. Skin grafting was scheduled but the retromammary space on the healthy side was dissected to the anterior border of the latissimus dorsi muscle, and the skin of the healthy side was used to cover the defect on the affected side. Consequently, the pendulous breast on the healthy side was elevated. This surgical technique provided an excellent aesthetic outcome without any skin problems, because autologous skin was used to fill the defect. Radiotherapy could subsequently be administered as scheduled. This procedure may be useful for elderly patients.
The small GTPase-effector proteins CDC42EP1-5/BORG1-5 interact reciprocally with CDC42 or the sep... more The small GTPase-effector proteins CDC42EP1-5/BORG1-5 interact reciprocally with CDC42 or the septin cytoskeleton. Here we show that, in the cerebellum, CDC42EP4 is exclusively expressed in Bergmann glia and localizes beneath specific membrane domains enwrapping dendritic spines of Purkinje cells. CDC42EP4 forms complexes with septin hetero-oligomers, which interact with a subset of glutamate transporter GLAST/EAAT1. In Cdc42ep4(-/-) mice, GLAST is dissociated from septins and is delocalized away from the parallel fibre-Purkinje cell synapses. The excitatory postsynaptic current exhibits a protracted decay time constant, reduced sensitivity to a competitive inhibitor of the AMPA-type glutamate receptors (γDGG) and excessive baseline inward current in response to a subthreshold dose of a nonselective inhibitor of the glutamate transporters/EAAT1-5 (DL-TBOA). Insufficient glutamate-buffering/clearance capacity in these mice manifests as motor coordination/learning defects, which are aggravated with subthreshold DL-TBOA. We propose that the CDC42EP4/septin-based glial scaffold facilitates perisynaptic localization of GLAST and optimizes the efficiency of glutamate-buffering and clearance.
The distribution of five NMDA receptor channel subunit mRNAs was examined in the mouse cerebellum... more The distribution of five NMDA receptor channel subunit mRNAs was examined in the mouse cerebellum from embryonic day 13 through postnatal day 56, by in situ hybridization with subunit-specific oligonucleotide probes. At postnatal days 21 and 56, each cerebellar neuron displayed differential expressions of the E subunit mRNAs. The granule cells showed hybridizing signals for the €1 and €3 subunit mRNAs, the molecular layer neurons for the €4 subunit mRNA, and the cerebellar nucleus neurons for the €1 and €4 subunit mRNAs, whereas the Purkinje cells did not express any E subunit mRNAs. At early postmitotic stages of development, the €2 subunit mRNA appeared in each cerebellar neuron, including the Purkinje cells, and the €4 subunit mRNA appeared in neurons of the molecular layer and the cerebellar nuclei. The expression patterns in the cerebellum altered drastically during the first 2 postnatal weeks: the €1 and €3 subunit mRNAs appeared in the granule cells and the cerebellar nucleus neurons, whereas the €2 subunit mRNA disappeared from each neuron and the signal levels of the €4 subunit mRNA decreased remarkably. In contrast to the differential expressions of the four E subunit mRNAs, intense signals for the 51 subunit mRNA were observed in each cerebellar neuron from early postmitotic stages through the mature stage. These findings suggest that anatomical organization of the E subunits is heterogeneous in the cerebellum both spatially and temporally, which would give rise to functional diversity of the NMDA receptor channel. D 1994 Wiley-Liss, Inc.
Background: One of the target enzymes of 5-fluorouracil (5-FU) is thymidylate synthetase (TS). Th... more Background: One of the target enzymes of 5-fluorouracil (5-FU) is thymidylate synthetase (TS). The DNA sequence of the TS gene includes either double or triple tandem 28-bp repeats within the promoter region, such that TS genotypes can be classified as homozygous 3R/3R, heterozygous 2R/3R or homozygous 2R/2R. Several recent studies have shown that TS genotype affects mRNA expression, with 3R/3R homozygotes showing higher TS mRNA expression compared to the other genotypes. Purpose: We analyzed the TS genotype and TS and dihydropyrimidine dehydrogenase (DPD) mRNA expression levels in 22 advanced gastric carcinoma patients, and analyzed results with respect to patient 5-FU chemosensitivity, as detected by the tetrazolium-based colorimetric (MTT) assay and survival outcome. Patients and Methods: Between September 2001 and April 2002, 22 Japanese patients with advanced gastric carcinoma were evaluated. Informed written consent was obtained from all patients and the study was approved by the ethical committee at our University Hospital. Fresh surgical specimens from carcinoma lesions were enzymatically dissociated and incubated with 5-FU at a concentration of 50 Ìg/ml for 48 hours to determine the inhibition rate as detected by MTT assay. Normal and tumor tissue and peripheral blood samples were collected and stored at -80AEC until assay for TS genotype and TS and DPD mRNA level. The TS genotype was assessed by PCR assay using peripheral monocytes, since monocyte genotypes represent the genotype of normal and tumor tissues. Quantification of TS and DPD mRNA levels was performed using real-time PCR. Survival outcome was assessed according to the disease-free survival period in cases with similar clinical backgrounds. Results: TS genotyping revealed 19 3R/3R homozygotes and 3 2R/3R heterozygotes. After analysis of normal and tumor tissues, samples from homozygote 3R/3R cases showed higher TS mRNA expression than heterozygote 2R/3R cases, which was statistically significant at p<0.05. We also observed a statistically significant correlation in TS mRNA levels between normal and tumor tissues, while no significant correlation was observed for DPD mRNA levels between normal and tumor tissues. While no relationship between 5-FU chemosensitivity and TS genotype or mRNA expression was observed, cases with high DPD mRNA expression were resistant to 5-FU and exhibited poor survival outcomes. Conclusion: While TS genotype affected TS mRNA expression in both normal and tumor tissues in advanced gastric cancer, there is no relationship between TS genotype or mRNA expression level and 5-FU chemosensitivity. Conclusion: Our finding, that DPD mRNA expression appears to be a factor in determining 5-FU chemosensitivity and the survival outcome of advanced gastric cancer patients, is comparable with previous reports.
The spontaneous recessive mutant mouse stargazer (stg) begins to show ataxia around postnatal day... more The spontaneous recessive mutant mouse stargazer (stg) begins to show ataxia around postnatal day 14 and display a severe impairment in the acquisition of classical eyeblink conditioning in adulthood. These abnormalities have been attributed to the specific reduction in brain-derived neurotrophic factor (BDNF) and the subsequent defect in TrkB receptor signaling in cerebellar granule cells (GCs). In the stg mutant cerebellum, we found that EPSCs at mossy fiber (MF) to GC synapses are devoid of the fast component mediated by AMPA-type glutamate receptors despite the normal slow component mediated by NMDA receptors. The sensitivity of stg mutant GCs to exogenously applied AMPA was greatly reduced, whereas that to NMDA was unchanged. Glutamate release from MF terminals during synaptic transmission to GCs appeared normal. By contrast, AMPA receptor-mediated EP-SCs were normal in CA1 pyramidal cells of the stg mutant hippocampus. Thus, postsynaptic AMPA receptor function was selectively impaired in stg mutant GCs, although the transcription of four AMPA receptor subunit genes in the stg GC was comparable to the wild-type GC. We also examined the cerebellum of BDNF knockout mice and found that their MF-GC synapses had a normal AMPA receptor-mediated EPSC component. Thus, the impaired AMPA receptor function in the stg mutant GC is not likely to result from the reduced BDNF-TrkB signaling. These results suggest that the defect in MF to GC synaptic transmission is a major factor that causes the cerebellar dysfunction in the stg mutant mouse.
Effects of atrial natriuretic peptides (ANP) on Type II cells of the rat lung were examined, usin... more Effects of atrial natriuretic peptides (ANP) on Type II cells of the rat lung were examined, using autoradiographic and morphometric techniques. The injection of an excess of ANP, together with [125I]ANP, significantly inhibited the uptake of radioactive ANP in the lung tissue. Following autoradiography, silver grains of [125I]ANP labelled Type II cells, endothelial cells and smooth muscle cells of vessels, bronchi and bronchioles. As for morphometric changes in subcellular structures of Type II cells after the injection of ANP, the volume and surface densities of the rough endoplasmic reticulum increased at 15 minutes, while those of the Golgi complex increased from 5 minutes, peaking at 30 minutes. At 15 minutes the volume and surface densities of mitochondria significantly increased. The volume and surface densities of multivesicular bodies with an electron-dense matrix also increased from 15 minutes after the injection. Lamellar bodies showed decreased volume and surface densities at 15 minutes whereas the densities showed an increase at 30 minutes and were higher at 60 minutes than those in the control. These results suggest that Type II cells provide a binding site for ANP which facilitates the production of lamellar bodies in addition to their secretion.
Gan to Kagaku Ryoho Cancer Chemotherapy, Nov 1, 2010
A 51-year-old postmenopausal woman was diagnosed as having adenocarcinoma (gastric cancer type 4)... more A 51-year-old postmenopausal woman was diagnosed as having adenocarcinoma (gastric cancer type 4) from gastric biopsy by upper endoscopy. Her chief complaint was abdominal dilatation. Meanwhile, a breast CT suggested tumor in her left breast and was diagnosed as an invasive lobular carcinoma based on a core needle biopsy. After gastric biopsy, tissues are stained by ER and PgR in immunohistochemistry. The diagnosis was modified from gastric cancer to T2N1M1, stage IV left breast cancer, accompanied by a treatment. Chemotherapy with EC 6 course consisted of a weekly PTX 4 course (epirubicin, cyclophosphamide-weekly paclitaxel) was performed. After the chemotherapy, breast mass, ascites and tumor marker were dramatically improved. Then hormonal therapy was administered. She passed away 2 and 1/2 years after her first visit to the hospital. Metastatic gastric tumors simulating type 4 advanced gastric cancer (MGTS type 4) and invasive lobular carcinoma are known to have an unfavorable prognosis. There is no doubt, however, that the multidisciplinary treatments have brought a satisfaction to her and family. We should keep in mind a possibility of gastric metastasis of breast cancer, when consulting a female patient with gastric cancer type 4.
The glutamate receptor ␦2 (GluR␦2) subunit is selectively expressed in cerebellar Purkinje cells ... more The glutamate receptor ␦2 (GluR␦2) subunit is selectively expressed in cerebellar Purkinje cells and plays an important role in cerebellar long-term depression, motor learning, motor coordination, and synapse development. We identified a novel GluR␦2-interacting protein, named Delphilin, that contains a single PDZ domain and formin homology (FH) domains FH1 and FH2 plus coiled-coil structure. As far as we know, this is the first reported protein that contains both PDZ and FH domains. Yeast two-hybrid and surface plasmon resonance (SPR) analyses indicated that Delphilin interacts with the GluR␦2 C terminus via its PDZ domain. This was also supported by coimmunoprecipitation experiments using a heterologous expression system in mammalian cells. Yeast cell and SPR analyses also demonstrated the possibility that the FH1 proline-rich region of Delphilin interacts with profilin, an actin-binding protein, and with the Src homology 3 domain of neuronal Src protein ty-rosine kinase. In situ hybridization demonstrated the highest expression of Delphilin mRNA in Purkinje cells. Delphilin polypeptide was highly enriched in the synaptosomal membrane fraction of the cerebellum and coimmunoprecipitated with the GluR␦2 subunit. The post-embedding immunogold technique demonstrated that Delphilin is selectively localized at the postsynaptic junction site of the parallel fiber-Purkinje cell synapse and colocalized with GluR␦2. Thus, Delphilin is a postsynaptic scaffolding protein at the parallel fiber-Purkinje cell synapse, where it may serve to link GluR␦2 with actin cytoskeleton and various signaling molecules.
A comprehensive search for DNA methylated genes identified candidate tumor suppressor genes that ... more A comprehensive search for DNA methylated genes identified candidate tumor suppressor genes that have been proven to be involved in the apoptotic process of the p53 pathway. In this study, we investigated p53 mutation in relation to such epigenetic alteration in primary gastric cancer. The methylation profiles of the 3 genes: PGP9.5, NMDAR2B, and CCNA1, which are involved in the p53 tumor suppressor pathway in combination with p53 mutation were examined in 163 primary gastric cancers. The effect of epigenetic reversion in combination with chemotherapeutic drugs on apoptosis was also assessed according to the tumor p53 mutation status. p53 gene mutations were found in 44 primary gastric tumors (27%), and super-high methylation of any of the 3 genes was only found in cases with wild type p53. Higher p53 pathway aberration was found in cases with male gender (p = 0.003), intestinal type (p = 0.005), and non-infiltrating type (p = 0.001). The p53 pathway aberration group exhibited less ...
To develop a cell type-specific and temporal regulation system of gene targeting in the cerebellu... more To develop a cell type-specific and temporal regulation system of gene targeting in the cerebellum, we used the NMDA-type glutamate receptor GluRe3 subunit gene and Cre recombinase- progesterone receptor fusion (CrePR) gene in combination. Injection of the CrePR gene placed under the control of the 10 kb 59 region of the GluRe3 gene into C57BL/6 eggs yielded the ECP25 line
The Journal of Neuroscience : The Official Journal of the Society for Neuroscience
Climbing fiber (CF) synapse formation onto cerebellar Purkinje cells (PCs) is critically dependen... more Climbing fiber (CF) synapse formation onto cerebellar Purkinje cells (PCs) is critically dependent on the synaptogenesis from parallel fibers (PFs), the other input to PCs. Previous studies revealed that deletion of the glutamate receptor ␦2 subunit (GluR␦2) gene results in persistent multiple CF innervation of PCs with impaired PF synaptogenesis, whereas mutation of the metabotropic glutamate receptor subtype 1 (mGluR1) gene causes multiple CF innervation with normal PF synaptogenesis. We demonstrate that atypical CF-mediated EPSCs (CF-EPSCs) with slow rise times and small amplitudes coexisted with typical CF-EPSCs with fast rise times and large amplitudes in PCs from GluR␦2 mutant cerebellar slices. CF-EPSCs in mGluR1 mutant and wild-type PCs had fast rise times. Atypical slow CF responses of GluR␦2 mutant PCs were associated with voltagedependent Ca 2ϩ signals that were confined to PC distal dendrites. In the wild-type and mGluR1 mutant PCs, CF-induced Ca 2ϩ signals involved both proximal and distal dendrites. Morphologically, CFs of GluR␦2 mutant mice extended to the superficial regions of the molecular layer, whereas those of wildtype and mGluR1 mutant mice did not innervate the superficial one-fifth of the molecular layer. It is therefore likely that surplus CFs of GluR␦2 mutant mice form ectopic synapses onto distal dendrites, whereas those of wild-type and mGluR1 mutant mice innervate proximal dendrites. These findings suggest that GluR␦2 is required for consolidating PF synapses and restricting CF synapses to the proximal dendrites, whereas the mGluR1-signaling pathway does not affect PF synaptogenesis but is involved in eliminating surplus CF synapses at the proximal dendrites.
Backgrounds The objectives of this study were to delineate differences in the morphologic charact... more Backgrounds The objectives of this study were to delineate differences in the morphologic characteristics of reflux esophagitis between a rat gastroesophageal reflux (GER) model and a duodenoesophageal reflux (DER) model and to evaluate the effects of H2-receptor antagonists on morphologic characteristics of reflux esophagitis in DER model. Methods Wistar rats were divided into 3 groups: GER model group (Group G), DER model group (Group D), and control group (Group C). Rats in each group were sacrificed 1 or 12 weeks after surgery. Intraesophageal pH was measured, and the excised esophagus was examined macroscopically and histologically. Subgroups of rats in Group D were given famotidine (10 mg/kg) or lafutidine (30 mg/kg) orally once daily for 1 week after surgery. The rats were then sacrificed, and histological findings were compared. Results Intraesophageal pH was significantly lower in Group G than in Group C. At 12 weeks, the epithelium of the lower esophagus in Groups G and D was significantly thicker than that in Group C and showed remarkable hyperplastic changes in Group D. The thickness of the epithelium in Group D ? famotidine did not differ significantly from that in Group D. In contrast, the epithelium was significantly thinner in Group D ? lafutidine than in Group D.
The Journal of Neuroscience : The Official Journal of the Society for Neuroscience
Phospholipase C (PLC) beta4, one of the four isoforms of PLCbetas, is the sole isoform expressed ... more Phospholipase C (PLC) beta4, one of the four isoforms of PLCbetas, is the sole isoform expressed in the mouse ventral posterolateral thalamic nucleus (VPL), a key station in pain processing. The mouse thalamus also has been shown to express a high level of metabotropic glutamate receptor type 1 (mGluR1), which stimulates PLCbetas through activation of Galphaq/11 protein. It is therefore expected that the thalamic mGluR1-PLCbeta4 cascade may play a functional role in nociceptive transmission. To test this hypothesis, we first studied behavioral responses to various nociceptive stimuli in PLCbeta4 knock-out mice. We performed the formalin test and found no difference in the pain behavior in the first phase of the formalin test, which is attributed to acute nociception, between PLCbeta4 knock-out and wild-type mice. Consistent with this result, acute pain responses in the hot plate and tail flick tests were also unaffected in the PLCbeta4 knock-out mice. However, the nociceptive behavi...
In the row NAC, the values in column 2yOS (%) for the classification DCF should be 100 instead of... more In the row NAC, the values in column 2yOS (%) for the classification DCF should be 100 instead of 84 and the values of 1yOS (%), 2yOS (%) for the classification FP should be 97 and 87 instead of 92 and 78, respectively.
Background Our aim in this study is to know whether clinical outcomes are improved by neoadjuvant... more Background Our aim in this study is to know whether clinical outcomes are improved by neoadjuvant chemotherapy (NAC) using Docetaxel/CDDP/5-FU (DCF) as compared to NAC using 5-FU/CDDP (FP). Methods Thirty-eight patients who underwent DCF NAC in cStage II/III esophageal squamous cell carcinoma (ESCC) were compared with the 41 counterparts treated by FP NAC. Docetaxel and CDDP were both given to 70-75 mg/m 2 with concurrent 5-FU at 750 mg/m 2 in 3 cycles. Median follow-up term of DCF NAC reached 27 months. Results In DCF NAC, grade 3 adverse effects were recognized in 97 %, and completion rate of the DCF NAC was 86 %. In terms of PR ? CR rate, DCF NAC was better (87 %) than FP NAC (59 %) (p = 0.005). Five year progression-free survival (PFS) and overall survival (OS) of FP NAC were 32 and 69 %, respectively, and OS of FP NAC was excellent putatively due to adoption of definitive chemoradiation therapy for recurrent diseases. Furthermore, survival was in favor of DCF NAC as compared to FP NAC for OS (p = 0.02) and PFS (p = 0.10), while R0 resection rate was similar. The 1st multivariate prognostic analysis among all cases with NAC revealed that significant factors were resectability and NAC modality for OS and PFS. We then performed the 2nd stage multivariate prognostic analysis limited to R0 cases including pathologic factors, which again identified DCF NAC modality as an independent prognostic factor. Conclusion DCF NAC for ESCC demonstrated high response rates, and may improve patient survival with acceptable feasibility.
The distributions of five NMDA receptor channel subunit mRNAs in the mouse forebrain at postnatal... more The distributions of five NMDA receptor channel subunit mRNAs in the mouse forebrain at postnatal day 21 were semiquantitatively examined by in situ hybridization with subunitspecific oligonucleotide probes. In contrast to ubiquitous distribution of the (1 subunit mRNA throughout the forebrain, distributions of four E subunit mRNAs were highly variable from nucleus to nucleus. The telencephalon (except for the septum) expressed the €1 and €2 subunit mRNAs. Various combinations of the €1, €2, €3, and €4 subunit mRNAs were present in different nuclei of the septum, the olfactory bulb, and the thalamus. In the hypothalamus, the suprachiasmatic nucleus expressed distinct signals for the €3 subunit mRNA alone, whereas other nuclei showed faint signals for the el, €2, and €4 subunit mRNAs. Moreover, different signal levels of the E subunit mRNAs were found in various regions. The hippocampal CA1 region expressed higher signals for the €1 and €2 subunit mRNAs than the CA3 region and the dentate gyrus. In the cerebral cortex, signal levels of the €1 subunit mRNA were higher in the laminae 111111, IV, and VI than the lamina V, whereas those of the €2 subunit mRNA were highest in laminae IIiIII and lowest in laminae IV and V. Different signal levels between the €1 and € 2 subunit mRNAs were also discerned in the amygdala, the caudate-putamen, and the thalamus. The distinct anatomical distributions and differential signal levels of the E subunit mRNAs strongly suggest different subunit organizations of the NMDA receptor channel in different forebrain neurons, which may result in functional diversity of the channel in vivo.
ABSTRACT Hierarchical State Transition Matrix (HSTM) is a table-based modeling language for devel... more ABSTRACT Hierarchical State Transition Matrix (HSTM) is a table-based modeling language for developing designs of software systems. Although widely used and adopted by (particularly Japanese) software industry, there is still lack of mechanized formal verification supports for conducting rigorous and automatic analysis to improve reliability of HSTM designs. In this paper, we first present a formalization of HSTM designs as state transition systems. Consequentially, based on this formalization, we propose a symbolic encoding approach, through which correctness of a HSTM design with respect to LTL properties could be represented as Bounded Model Checking (BMC) problems that could be determined by Satisfiability Modulo Theories (SMT) solving. We have implemented our encoding approach in a tool called Garakabu2 with the state-of-the-art SMT solver CVC3 as its back-ended solver. Furthermore, in our preliminary experiments, a conceptually simple but steadily effective way of accelerating SMT solving for HSTM designs is investigated and reported.
Gan to Kagaku Ryoho Cancer Chemotherapy, Nov 1, 2012
Patient 1 was a 63-year-old woman whose chief complaint was a mass in the left breast. Physical e... more Patient 1 was a 63-year-old woman whose chief complaint was a mass in the left breast. Physical examination revealed an inverted left nipple, a very large mass on the anterior aspect of the sternum, and erythema. Because the tumor had directly invaded the sternum, T4cN3M0, stage IIIC breast cancer was diagnosed. The patient preoperatively received chemotherapy with 6 courses of FEC100 (5-fluorouracil, epirubicin, and cyclophosphamide) and 5 courses of nanoparticle albumin -bound paclitaxel (260 mg/m2), which enabled a partial response. Patient 2 was an 83-year-old woman whose chief complaint was a mass in the upper internal and external quadrants of the right breast measuring 20×15 cm and erythema. The mass was accompanied by enlarged right axillary lymph nodes(T4bN1M0, stage IIIB breast cancer). Both patients underwent core needle biopsy of the skin and breast masses. They were both diagnosed with invasive, lobular, triple-negative breast cancer (estrogen receptor negative, progesterone receptor negative, human epidermal growth factor receptor 2 negative). The surgical resection line was drawn to include the extensive skin invasion, and mastectomy and axillary dissection were performed. Skin grafting was scheduled but the retromammary space on the healthy side was dissected to the anterior border of the latissimus dorsi muscle, and the skin of the healthy side was used to cover the defect on the affected side. Consequently, the pendulous breast on the healthy side was elevated. This surgical technique provided an excellent aesthetic outcome without any skin problems, because autologous skin was used to fill the defect. Radiotherapy could subsequently be administered as scheduled. This procedure may be useful for elderly patients.
The small GTPase-effector proteins CDC42EP1-5/BORG1-5 interact reciprocally with CDC42 or the sep... more The small GTPase-effector proteins CDC42EP1-5/BORG1-5 interact reciprocally with CDC42 or the septin cytoskeleton. Here we show that, in the cerebellum, CDC42EP4 is exclusively expressed in Bergmann glia and localizes beneath specific membrane domains enwrapping dendritic spines of Purkinje cells. CDC42EP4 forms complexes with septin hetero-oligomers, which interact with a subset of glutamate transporter GLAST/EAAT1. In Cdc42ep4(-/-) mice, GLAST is dissociated from septins and is delocalized away from the parallel fibre-Purkinje cell synapses. The excitatory postsynaptic current exhibits a protracted decay time constant, reduced sensitivity to a competitive inhibitor of the AMPA-type glutamate receptors (γDGG) and excessive baseline inward current in response to a subthreshold dose of a nonselective inhibitor of the glutamate transporters/EAAT1-5 (DL-TBOA). Insufficient glutamate-buffering/clearance capacity in these mice manifests as motor coordination/learning defects, which are aggravated with subthreshold DL-TBOA. We propose that the CDC42EP4/septin-based glial scaffold facilitates perisynaptic localization of GLAST and optimizes the efficiency of glutamate-buffering and clearance.
The distribution of five NMDA receptor channel subunit mRNAs was examined in the mouse cerebellum... more The distribution of five NMDA receptor channel subunit mRNAs was examined in the mouse cerebellum from embryonic day 13 through postnatal day 56, by in situ hybridization with subunit-specific oligonucleotide probes. At postnatal days 21 and 56, each cerebellar neuron displayed differential expressions of the E subunit mRNAs. The granule cells showed hybridizing signals for the €1 and €3 subunit mRNAs, the molecular layer neurons for the €4 subunit mRNA, and the cerebellar nucleus neurons for the €1 and €4 subunit mRNAs, whereas the Purkinje cells did not express any E subunit mRNAs. At early postmitotic stages of development, the €2 subunit mRNA appeared in each cerebellar neuron, including the Purkinje cells, and the €4 subunit mRNA appeared in neurons of the molecular layer and the cerebellar nuclei. The expression patterns in the cerebellum altered drastically during the first 2 postnatal weeks: the €1 and €3 subunit mRNAs appeared in the granule cells and the cerebellar nucleus neurons, whereas the €2 subunit mRNA disappeared from each neuron and the signal levels of the €4 subunit mRNA decreased remarkably. In contrast to the differential expressions of the four E subunit mRNAs, intense signals for the 51 subunit mRNA were observed in each cerebellar neuron from early postmitotic stages through the mature stage. These findings suggest that anatomical organization of the E subunits is heterogeneous in the cerebellum both spatially and temporally, which would give rise to functional diversity of the NMDA receptor channel. D 1994 Wiley-Liss, Inc.
Background: One of the target enzymes of 5-fluorouracil (5-FU) is thymidylate synthetase (TS). Th... more Background: One of the target enzymes of 5-fluorouracil (5-FU) is thymidylate synthetase (TS). The DNA sequence of the TS gene includes either double or triple tandem 28-bp repeats within the promoter region, such that TS genotypes can be classified as homozygous 3R/3R, heterozygous 2R/3R or homozygous 2R/2R. Several recent studies have shown that TS genotype affects mRNA expression, with 3R/3R homozygotes showing higher TS mRNA expression compared to the other genotypes. Purpose: We analyzed the TS genotype and TS and dihydropyrimidine dehydrogenase (DPD) mRNA expression levels in 22 advanced gastric carcinoma patients, and analyzed results with respect to patient 5-FU chemosensitivity, as detected by the tetrazolium-based colorimetric (MTT) assay and survival outcome. Patients and Methods: Between September 2001 and April 2002, 22 Japanese patients with advanced gastric carcinoma were evaluated. Informed written consent was obtained from all patients and the study was approved by the ethical committee at our University Hospital. Fresh surgical specimens from carcinoma lesions were enzymatically dissociated and incubated with 5-FU at a concentration of 50 Ìg/ml for 48 hours to determine the inhibition rate as detected by MTT assay. Normal and tumor tissue and peripheral blood samples were collected and stored at -80AEC until assay for TS genotype and TS and DPD mRNA level. The TS genotype was assessed by PCR assay using peripheral monocytes, since monocyte genotypes represent the genotype of normal and tumor tissues. Quantification of TS and DPD mRNA levels was performed using real-time PCR. Survival outcome was assessed according to the disease-free survival period in cases with similar clinical backgrounds. Results: TS genotyping revealed 19 3R/3R homozygotes and 3 2R/3R heterozygotes. After analysis of normal and tumor tissues, samples from homozygote 3R/3R cases showed higher TS mRNA expression than heterozygote 2R/3R cases, which was statistically significant at p<0.05. We also observed a statistically significant correlation in TS mRNA levels between normal and tumor tissues, while no significant correlation was observed for DPD mRNA levels between normal and tumor tissues. While no relationship between 5-FU chemosensitivity and TS genotype or mRNA expression was observed, cases with high DPD mRNA expression were resistant to 5-FU and exhibited poor survival outcomes. Conclusion: While TS genotype affected TS mRNA expression in both normal and tumor tissues in advanced gastric cancer, there is no relationship between TS genotype or mRNA expression level and 5-FU chemosensitivity. Conclusion: Our finding, that DPD mRNA expression appears to be a factor in determining 5-FU chemosensitivity and the survival outcome of advanced gastric cancer patients, is comparable with previous reports.
The spontaneous recessive mutant mouse stargazer (stg) begins to show ataxia around postnatal day... more The spontaneous recessive mutant mouse stargazer (stg) begins to show ataxia around postnatal day 14 and display a severe impairment in the acquisition of classical eyeblink conditioning in adulthood. These abnormalities have been attributed to the specific reduction in brain-derived neurotrophic factor (BDNF) and the subsequent defect in TrkB receptor signaling in cerebellar granule cells (GCs). In the stg mutant cerebellum, we found that EPSCs at mossy fiber (MF) to GC synapses are devoid of the fast component mediated by AMPA-type glutamate receptors despite the normal slow component mediated by NMDA receptors. The sensitivity of stg mutant GCs to exogenously applied AMPA was greatly reduced, whereas that to NMDA was unchanged. Glutamate release from MF terminals during synaptic transmission to GCs appeared normal. By contrast, AMPA receptor-mediated EP-SCs were normal in CA1 pyramidal cells of the stg mutant hippocampus. Thus, postsynaptic AMPA receptor function was selectively impaired in stg mutant GCs, although the transcription of four AMPA receptor subunit genes in the stg GC was comparable to the wild-type GC. We also examined the cerebellum of BDNF knockout mice and found that their MF-GC synapses had a normal AMPA receptor-mediated EPSC component. Thus, the impaired AMPA receptor function in the stg mutant GC is not likely to result from the reduced BDNF-TrkB signaling. These results suggest that the defect in MF to GC synaptic transmission is a major factor that causes the cerebellar dysfunction in the stg mutant mouse.
Effects of atrial natriuretic peptides (ANP) on Type II cells of the rat lung were examined, usin... more Effects of atrial natriuretic peptides (ANP) on Type II cells of the rat lung were examined, using autoradiographic and morphometric techniques. The injection of an excess of ANP, together with [125I]ANP, significantly inhibited the uptake of radioactive ANP in the lung tissue. Following autoradiography, silver grains of [125I]ANP labelled Type II cells, endothelial cells and smooth muscle cells of vessels, bronchi and bronchioles. As for morphometric changes in subcellular structures of Type II cells after the injection of ANP, the volume and surface densities of the rough endoplasmic reticulum increased at 15 minutes, while those of the Golgi complex increased from 5 minutes, peaking at 30 minutes. At 15 minutes the volume and surface densities of mitochondria significantly increased. The volume and surface densities of multivesicular bodies with an electron-dense matrix also increased from 15 minutes after the injection. Lamellar bodies showed decreased volume and surface densities at 15 minutes whereas the densities showed an increase at 30 minutes and were higher at 60 minutes than those in the control. These results suggest that Type II cells provide a binding site for ANP which facilitates the production of lamellar bodies in addition to their secretion.
Gan to Kagaku Ryoho Cancer Chemotherapy, Nov 1, 2010
A 51-year-old postmenopausal woman was diagnosed as having adenocarcinoma (gastric cancer type 4)... more A 51-year-old postmenopausal woman was diagnosed as having adenocarcinoma (gastric cancer type 4) from gastric biopsy by upper endoscopy. Her chief complaint was abdominal dilatation. Meanwhile, a breast CT suggested tumor in her left breast and was diagnosed as an invasive lobular carcinoma based on a core needle biopsy. After gastric biopsy, tissues are stained by ER and PgR in immunohistochemistry. The diagnosis was modified from gastric cancer to T2N1M1, stage IV left breast cancer, accompanied by a treatment. Chemotherapy with EC 6 course consisted of a weekly PTX 4 course (epirubicin, cyclophosphamide-weekly paclitaxel) was performed. After the chemotherapy, breast mass, ascites and tumor marker were dramatically improved. Then hormonal therapy was administered. She passed away 2 and 1/2 years after her first visit to the hospital. Metastatic gastric tumors simulating type 4 advanced gastric cancer (MGTS type 4) and invasive lobular carcinoma are known to have an unfavorable prognosis. There is no doubt, however, that the multidisciplinary treatments have brought a satisfaction to her and family. We should keep in mind a possibility of gastric metastasis of breast cancer, when consulting a female patient with gastric cancer type 4.
The glutamate receptor ␦2 (GluR␦2) subunit is selectively expressed in cerebellar Purkinje cells ... more The glutamate receptor ␦2 (GluR␦2) subunit is selectively expressed in cerebellar Purkinje cells and plays an important role in cerebellar long-term depression, motor learning, motor coordination, and synapse development. We identified a novel GluR␦2-interacting protein, named Delphilin, that contains a single PDZ domain and formin homology (FH) domains FH1 and FH2 plus coiled-coil structure. As far as we know, this is the first reported protein that contains both PDZ and FH domains. Yeast two-hybrid and surface plasmon resonance (SPR) analyses indicated that Delphilin interacts with the GluR␦2 C terminus via its PDZ domain. This was also supported by coimmunoprecipitation experiments using a heterologous expression system in mammalian cells. Yeast cell and SPR analyses also demonstrated the possibility that the FH1 proline-rich region of Delphilin interacts with profilin, an actin-binding protein, and with the Src homology 3 domain of neuronal Src protein ty-rosine kinase. In situ hybridization demonstrated the highest expression of Delphilin mRNA in Purkinje cells. Delphilin polypeptide was highly enriched in the synaptosomal membrane fraction of the cerebellum and coimmunoprecipitated with the GluR␦2 subunit. The post-embedding immunogold technique demonstrated that Delphilin is selectively localized at the postsynaptic junction site of the parallel fiber-Purkinje cell synapse and colocalized with GluR␦2. Thus, Delphilin is a postsynaptic scaffolding protein at the parallel fiber-Purkinje cell synapse, where it may serve to link GluR␦2 with actin cytoskeleton and various signaling molecules.
A comprehensive search for DNA methylated genes identified candidate tumor suppressor genes that ... more A comprehensive search for DNA methylated genes identified candidate tumor suppressor genes that have been proven to be involved in the apoptotic process of the p53 pathway. In this study, we investigated p53 mutation in relation to such epigenetic alteration in primary gastric cancer. The methylation profiles of the 3 genes: PGP9.5, NMDAR2B, and CCNA1, which are involved in the p53 tumor suppressor pathway in combination with p53 mutation were examined in 163 primary gastric cancers. The effect of epigenetic reversion in combination with chemotherapeutic drugs on apoptosis was also assessed according to the tumor p53 mutation status. p53 gene mutations were found in 44 primary gastric tumors (27%), and super-high methylation of any of the 3 genes was only found in cases with wild type p53. Higher p53 pathway aberration was found in cases with male gender (p = 0.003), intestinal type (p = 0.005), and non-infiltrating type (p = 0.001). The p53 pathway aberration group exhibited less ...
To develop a cell type-specific and temporal regulation system of gene targeting in the cerebellu... more To develop a cell type-specific and temporal regulation system of gene targeting in the cerebellum, we used the NMDA-type glutamate receptor GluRe3 subunit gene and Cre recombinase- progesterone receptor fusion (CrePR) gene in combination. Injection of the CrePR gene placed under the control of the 10 kb 59 region of the GluRe3 gene into C57BL/6 eggs yielded the ECP25 line
The Journal of Neuroscience : The Official Journal of the Society for Neuroscience
Climbing fiber (CF) synapse formation onto cerebellar Purkinje cells (PCs) is critically dependen... more Climbing fiber (CF) synapse formation onto cerebellar Purkinje cells (PCs) is critically dependent on the synaptogenesis from parallel fibers (PFs), the other input to PCs. Previous studies revealed that deletion of the glutamate receptor ␦2 subunit (GluR␦2) gene results in persistent multiple CF innervation of PCs with impaired PF synaptogenesis, whereas mutation of the metabotropic glutamate receptor subtype 1 (mGluR1) gene causes multiple CF innervation with normal PF synaptogenesis. We demonstrate that atypical CF-mediated EPSCs (CF-EPSCs) with slow rise times and small amplitudes coexisted with typical CF-EPSCs with fast rise times and large amplitudes in PCs from GluR␦2 mutant cerebellar slices. CF-EPSCs in mGluR1 mutant and wild-type PCs had fast rise times. Atypical slow CF responses of GluR␦2 mutant PCs were associated with voltagedependent Ca 2ϩ signals that were confined to PC distal dendrites. In the wild-type and mGluR1 mutant PCs, CF-induced Ca 2ϩ signals involved both proximal and distal dendrites. Morphologically, CFs of GluR␦2 mutant mice extended to the superficial regions of the molecular layer, whereas those of wildtype and mGluR1 mutant mice did not innervate the superficial one-fifth of the molecular layer. It is therefore likely that surplus CFs of GluR␦2 mutant mice form ectopic synapses onto distal dendrites, whereas those of wild-type and mGluR1 mutant mice innervate proximal dendrites. These findings suggest that GluR␦2 is required for consolidating PF synapses and restricting CF synapses to the proximal dendrites, whereas the mGluR1-signaling pathway does not affect PF synaptogenesis but is involved in eliminating surplus CF synapses at the proximal dendrites.
Backgrounds The objectives of this study were to delineate differences in the morphologic charact... more Backgrounds The objectives of this study were to delineate differences in the morphologic characteristics of reflux esophagitis between a rat gastroesophageal reflux (GER) model and a duodenoesophageal reflux (DER) model and to evaluate the effects of H2-receptor antagonists on morphologic characteristics of reflux esophagitis in DER model. Methods Wistar rats were divided into 3 groups: GER model group (Group G), DER model group (Group D), and control group (Group C). Rats in each group were sacrificed 1 or 12 weeks after surgery. Intraesophageal pH was measured, and the excised esophagus was examined macroscopically and histologically. Subgroups of rats in Group D were given famotidine (10 mg/kg) or lafutidine (30 mg/kg) orally once daily for 1 week after surgery. The rats were then sacrificed, and histological findings were compared. Results Intraesophageal pH was significantly lower in Group G than in Group C. At 12 weeks, the epithelium of the lower esophagus in Groups G and D was significantly thicker than that in Group C and showed remarkable hyperplastic changes in Group D. The thickness of the epithelium in Group D ? famotidine did not differ significantly from that in Group D. In contrast, the epithelium was significantly thinner in Group D ? lafutidine than in Group D.
The Journal of Neuroscience : The Official Journal of the Society for Neuroscience
Phospholipase C (PLC) beta4, one of the four isoforms of PLCbetas, is the sole isoform expressed ... more Phospholipase C (PLC) beta4, one of the four isoforms of PLCbetas, is the sole isoform expressed in the mouse ventral posterolateral thalamic nucleus (VPL), a key station in pain processing. The mouse thalamus also has been shown to express a high level of metabotropic glutamate receptor type 1 (mGluR1), which stimulates PLCbetas through activation of Galphaq/11 protein. It is therefore expected that the thalamic mGluR1-PLCbeta4 cascade may play a functional role in nociceptive transmission. To test this hypothesis, we first studied behavioral responses to various nociceptive stimuli in PLCbeta4 knock-out mice. We performed the formalin test and found no difference in the pain behavior in the first phase of the formalin test, which is attributed to acute nociception, between PLCbeta4 knock-out and wild-type mice. Consistent with this result, acute pain responses in the hot plate and tail flick tests were also unaffected in the PLCbeta4 knock-out mice. However, the nociceptive behavi...
In the row NAC, the values in column 2yOS (%) for the classification DCF should be 100 instead of... more In the row NAC, the values in column 2yOS (%) for the classification DCF should be 100 instead of 84 and the values of 1yOS (%), 2yOS (%) for the classification FP should be 97 and 87 instead of 92 and 78, respectively.
Background Our aim in this study is to know whether clinical outcomes are improved by neoadjuvant... more Background Our aim in this study is to know whether clinical outcomes are improved by neoadjuvant chemotherapy (NAC) using Docetaxel/CDDP/5-FU (DCF) as compared to NAC using 5-FU/CDDP (FP). Methods Thirty-eight patients who underwent DCF NAC in cStage II/III esophageal squamous cell carcinoma (ESCC) were compared with the 41 counterparts treated by FP NAC. Docetaxel and CDDP were both given to 70-75 mg/m 2 with concurrent 5-FU at 750 mg/m 2 in 3 cycles. Median follow-up term of DCF NAC reached 27 months. Results In DCF NAC, grade 3 adverse effects were recognized in 97 %, and completion rate of the DCF NAC was 86 %. In terms of PR ? CR rate, DCF NAC was better (87 %) than FP NAC (59 %) (p = 0.005). Five year progression-free survival (PFS) and overall survival (OS) of FP NAC were 32 and 69 %, respectively, and OS of FP NAC was excellent putatively due to adoption of definitive chemoradiation therapy for recurrent diseases. Furthermore, survival was in favor of DCF NAC as compared to FP NAC for OS (p = 0.02) and PFS (p = 0.10), while R0 resection rate was similar. The 1st multivariate prognostic analysis among all cases with NAC revealed that significant factors were resectability and NAC modality for OS and PFS. We then performed the 2nd stage multivariate prognostic analysis limited to R0 cases including pathologic factors, which again identified DCF NAC modality as an independent prognostic factor. Conclusion DCF NAC for ESCC demonstrated high response rates, and may improve patient survival with acceptable feasibility.
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Papers by Masahiko Watanabe