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Biomedical subjects

H Yamamoto

Publications and source records attributed to H Yamamoto.

At least 109 records · Page 6Linked to original sources

Identification of differentially expressed genes involved in colorectal carcinogenesis using a cDNA microarray.

To identify candidate genes involved in human colorectal carcinogenesis, we constructed the gene expression profiles of 50 colorectal cancers (CRCs) and 12 normal colorectal epithelia using a cDNA microarray specially constructed for CRC. Hierarchical clustering analysis and principal component analysis could clearly distinguish the gene profiles of cancer tissues from those of normal tissues. Our results confirm there are indeed differences in gene expression between cancer and normal mucosa. Our cDNA microarray identified 22 up-regulated genes and 32 down-regulated genes in CRC. Many of these genes have been previously identified in relation to human carcinogenesis, 68% and 78%, respectively. Subsequent validation of selected genes by serial analysis of gene expression and reverse transcription polymerase chain reaction, demonstrated expression patterns that were almost identical to our microarray analysis. Using a four-fold larger sample relative to that used in our previous study, candidate genes involved in human colorectal carcinogenesis were reproducibly identified. Further studies of comprehensive gene expression using our technique may elucidate the mechanism of CRC tumorigenesis.

Cell Line, Tumor↗

Determination of indices of the corpus callosum associated with normal aging in Japanese individuals.

Indices of the corpus callosum with normal aging and their sex differences were elucidated using quantitative MRI. We studied 94 Japanese men (mean+/-SD 57.3+/-20.8 years, range 6-90 years) and 111 Japanese women (mean+/-SD 61.2+/-17.6 years, range 9-86 years) who had no intracranial lesions on MRI and no history of neurological illness. The widths of the rostrum, body and splenium, the anterior to posterior length, and the maximum height in the midsagittal image were selected for measurement. The Evans index, which is the relative ratio of lateral ventricle expansion, and the maximum width of the third ventricle in the axial image were also estimated for comparison. The widths of rostrum, body and splenium of the corpus callosum became thinner with age. Conversely, the anterior to posterior length and the maximum height of the corpus callosum increased with age. The ratio of the width of the body to the length of the corpus callosum and the ratio of the width of the body to the height of the corpus callosum are best correlated with age. No sex differences in regional size of corpus callosum, including these two ratios, were observed in any raw measures, although ventricular indices were larger in men than women. Evaluation of the ratio of the width of the body to its length and the ratio of the width of the body to its height may enable accurate estimation of normal or pathological changes of the corpus callosum. Aging and pathological atrophy of corpus callosum can be evaluated without any adjustment for gender.

Adolescent↗

Overexpression of leucocyte common antigen (LAR) P-subunit in thyroid carcinomas.

Protein tyrosine phosphatase (PTPase) dephosphorylation and protein tyrosine kinase (PTKs) phosphorylation of key signal transduction proteins may be regulated by extracellular signals, making PTPases important in the regulation of cell proliferation. Leucocyte common antigen (LAR), a receptor-like PTPase, consists of E-subunit, containing the cell adhesion molecule-like receptor region, and P-subunit specific for a short segment of the extracellular region, the transmembrane peptide, and two cytoplasmic PTPase domains. We produced a monoclonal antibody against the LAR P-subunit for immunohistochemical screening of LAR expression in normal and tumourous tissues. Gliomas and gastric, colorectal, lung, breast and prostate cancers showed weak and relatively infrequent expression. Intense and diffuse expression, however, was detected in 95% (227 out of 239) of thyroid carcinomas, but only 12% (22 out of 128) of adenomas and no cases of benign thyroid disease were immunopositive. In contrast to broad staining in carcinomas, LAR expression in thyroid adenomas was often found in small focal or locally invasive areas. Western blot analysis similarly detected LAR P-subunit protein in thyroid carcinomas, but not in normal tissues. We believe this to be the first demonstration of LAR overexpression in thyroid carcinoma and may help to elucidate the role of PTPases in the development of malignancy.

Antigens, CD↗

Essential Bacillus subtilis genes.

To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. Among approximately 4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. Another 79 genes were predicted to be essential. The vast majority of essential genes were categorized in relatively few domains of cell metabolism, with about half involved in information processing, one-fifth involved in the synthesis of cell envelope and the determination of cell shape and division, and one-tenth related to cell energetics. Only 4% of essential genes encode unknown functions. Most essential genes are present throughout a wide range of Bacteria, and almost 70% can also be found in Archaea and Eucarya. However, essential genes related to cell envelope, shape, division, and respiration tend to be lost from bacteria with small genomes. Unexpectedly, most genes involved in the Embden-Meyerhof-Parnas pathway are essential. Identification of unknown and unexpected essential genes opens research avenues to better understanding of processes that sustain bacterial life.

Bacillus subtilis↗

Synthesis of ethyl ( S)-4-chloro-3-hydroxybutanoate using fabG-homologues.

This paper is a report on the successful application of bioinformatics to enzyme screening. The synthesis of ethyl ( S)-4-chloro-3-hydroxybutanoate (ECHB) by asymmetric reduction of ethyl 4-chloroacetoacetate (ECAA) using fabG-homologues was studied. beta-Ketoacyl-acyl carrier protein reductases from both Escherichia coli and Bacillus subtilis, which are components of type II fatty acid synthase, could reduce ECAA to ( S)-ECHB with 94-98% ee. Furthermore, acetoacetyl-CoA reductases (ARs) from both Ralstonia eutropha and Zoogloea ramigera, whose genes are significantly similar to fabG genes and play a physiological role in the biosynthesis of poly-beta-3-hydroxybutyrate, could also catalyze the asymmetric reduction of ECAA to ( S)-ECHB with >99% ee. ( S)-ECHB was synthesized to 48.7 g/l with an optical purity of 99.8% ee, using recombinant E. coli cells coexpressing AR from R. eutropha and glucose dehydrogenase from B. subtilis for the regeneration of NADPH.

3-Oxoacyl-(Acyl-Carrier-Protein) Reductase↗

Mucoadhesive properties of carbopol or chitosan-coated liposomes and their effectiveness in the oral administration of calcitonin to rats.

Mucoadhesive liposomes were prepared by coating multilamellar liposomes with Carbopol (CP) in a similar manner to that used in the preparation of chitosan-coated liposomes (CS-Lip) previously reported [Takeuchi et al., Chem. Pharm. Bull. 42 (1994) 1954-1956; Pharm. Res. 13 (1996) 896-901]. The positively charged liposomes containing dipalmitoylphosphatidylcholine (DPPC) and stearylamine at an optimum formulation ratio (40:1) could be coated with CP without aggregation of the liposomal particles during the coating process. The mucoadhesive properties of both the resultant polymer-coated liposomes (CP-Lip and CS-Lip) and the positively or negatively charged noncoated liposomes (Non-Lip) were evaluated using the rat intestine. The order was CS-Lip> or = CP-Lip>positively charged Non-Lip>negatively charged Non-Lip. The adhesive property of CP-Lip decreased on increasing pH of the dispersing medium from pH 5.0 to 7.4, probably owing to the electric repulsion between CP-Lip and the mucus layer. The effectiveness of the liposomal formulations in oral administration of calcitonin was evaluated by measuring the change in the blood calcium concentration of rats. Administration of CP-Lip and CS-Lip containing calcitonin showed an enhanced and prolonged reduction in blood calcium concentration. The overall pharmacological effect of CP- and CS-Lips evaluated by means of the area under the plasma calcium concentration curve was 2.4 and 2.8 times higher than that of negatively and positively charged Non-Lips, respectively.

Acrylic Resins↗

Nucleotide sequences of the coat protein gene of potyviruses infecting Ornithogalum thyrsoides.

Species of three viral genera infecting Ornithogalum thyrsoides plants showing mosaic symptoms were identified using RT-PCR and degenerate universal primers for each viral genus. The DNA fragments obtained encoded the coat protein (CP) gene and were sequenced. The plants were found to be infected with one or other of three potyvirus species, one of them was Ornithogalum mosaic virus (OrMV). The other two viruses were previously unrecorded and were named Ornithogalum virus 2 (OV-2) and 3 (OV-3). Direct comparison and phylogenetic analysis with published OrMV isolates revealed that the CP of the three OrMV-like clones were more similar to Pterostylis virus Y (PtVY) than to OrMV. No carlavirus or potexvirus was isolated.

Base Sequence↗

Characterization of a new potyvirus isolated from Chinese artichoke in Japan.

Afilamentous virus was isolated in Japan from virus-infected Chinese artichoke showing mosaic symptoms. This virus was assigned to the genus Potyvirus, based on particle morphology and serological properties. The virus could be transmitted to several test plants but not to Perilla frutesence, the host plant of Perilla mottle virus. cDNA corresponding to the 3'-terminal 1675 nucleotides of the viral RNA, excluding the poly (A) tail, was cloned and sequenced. The amino acid sequence of the coat protein was different from those of 74 distinct potyviruses. Therefore, we propose that the new potyvirus should be designated as Chinese artichoke mosaic virus (ChAMV).

Capsid Proteins↗

Risk factors for asymptomatic atherosclerosis in Japanese type 2 diabetic patients without diabetic microvascular complications.

Atherosclerotic vascular diseases are frequently associated with diabetes mellitus. There has been increasing evidence showing that the atherosclerotic diseases in diabetic patients are distinct from diabetic microvascular complications as to their pathophysiology and epidemiology. However, we have no information on the prevalence of asymptomatic atherosclerosis in diabetic patients before the onset of microvascular diseases. In the present investigation, we aimed to evaluate risk factors for the atherosclerosis in type 2 diabetic patients without the microvascular diseases. For this purpose, we evaluated atherosclerotic change of carotid arteries in 125 Japanese type 2 diabetic patients who had neither atherosclerotic vascular diseases nor diabetic microvascular complications. When atherosclerotic change was defined as the mean intima-media thickness (IMT) of >/= 1.1 mm and/or the presence of plaque lesion, 50% of patients had atherosclerosis of the carotid arteries. Risk factors for the carotid atherosclerosis were age, low-density lipoprotein (LDL)-cholesterol, hypertension, and diabetes treatment. Age and LDL-cholesterol were associated with mean IMT. Age, diabetes treatment, LDL-cholesterol, and hypertension were positively associated with plaque lesion, while high-density lipoprotein (HDL)-cholesterol was negatively associated with it. Fasting plasma glucose, glycosylated hemoglobin (HbA(1c)), and known diabetes duration remained unassociated with any parameters of asymptomatic atherosclerosis of the carotid arteries. These results indicate that glycemic control is unrelated with asymptomatic atherosclerosis in type 2 diabetic patients without diabetic microvascular complications. Conventional risk factors and diabetes treatment are independently associated with atherosclerosis of the carotid arteries in these patients.

Aged↗

Delayed neuronal damage related to microglia proliferation after mild spinal cord compression injury.

In order to investigate the mechanism of delayed progressive or secondary neuronal damage after the spinal cord injury, we developed a mild-compression injury model in the rat thoracic spinal cord. Our compression device consists of a soft silicone point of contact to the dura, in order to prevent violent injury that may cause axonal tears or hemorrhages in the spinal cord. Since rats often assume a 'standing' posture, i.e. raising head with lifting their fore-limbs, damage to the thoracic spinal cord was evaluated by measuring the frequency of 'standing', which effectively indicates hind limb function. Twenty-four hours after compression by a 20 g weight for 10 or 20 min, the standing frequency of the injured rat was almost the same as that of sham animals that underwent laminectomy without compression. However, the standing frequency decreased with time; the frequency of standing at 72 h was approximately 30-50% that of sham animals. In the compressed spinal cord tissue, microglial cells, detected by lectin staining, proliferated with time. An enormous amount of microglia was observed at 48 and 72 h after compression, although only a small amount of cells were positive to lectin staining at 24 h after the compression. These results suggest that our mild-compression spinal cord injury model showed late-onset or delayed neuronal damage that may be related to pathological microglia proliferation.

Animals↗

Simultaneous stimulation of EP2 and EP4 is essential to the effect of prostaglandin E2 in chondrocyte differentiation.

OBJECTIVE: Prostaglandin E(2)(PGE(2)) has been reported to stimulate chondrocyte differentiation. However, the precise actions and signal transduction pathways of PGE(2)in cartilage are largely unknown. Our purpose is to identify which of the four PGE(2)receptor subtype(s), EP1-4, mediates the action of PGE(2)on chondrocyte differentiation. DESIGN: We used primary chondrocytes derived from the resting zone of rat rib cartilage. The effects on chondrocyte differentiation were assessed by measuring the Alcian blue-stainable proteoglycan content and the expression levels of type II collagen mRNA by Northern blot analysis. The expression of the four PGE(2)receptor subtypes in rat primary chondrocytes was examined by reverse transcription-polymerase chain reaction. RESULTS: PGE(2)stimulated the accumulation of proteoglycan and up-regulated the expression of type II collagen mRNA in primary chondrocytes. Dibutyryl cAMP, a cell-permeable analog of cAMP, an important intracellular mediator of PGE(2)signaling, also enhanced the expression of type II collagen mRNA and proteoglycan accumulation in chondrocytes. No EP agonist alone induced the expression of type II collagen mRNA. However, simultaneous administration of EP2 and EP4 agonists at high concentrations cooperatively induced the expression of type II collagen mRNA, mimicking the PGE(2)effect. The simultaneous stimulation of EP2 and EP4 also cooperatively enhanced proteoglycan accumulation and intracellular cAMP production. Moreover, an EP4 antagonist partially blocked the stimulatory actions of PGE(2)on the expression of type II collagen mRNA. CONCLUSION: These results suggest that simultaneous stimulation of EP2 and EP4 is necessary and sufficient to elicit the effect of PGE(2)on rat primary chondrocyte differentiation.

Animals↗

Amended biochemical characteristics and phylogenetic position of Treponema medium.

Umemoto et al. (1997, Int J Syst Bacteriol 47, pp. 67-72) proposed spirochete strain G7201, isolated from the periodontal pocket of an adult patient, as a new species, Treponema medium. They deposited this strain in the American Type Culture Collection (ATCC) as type strain ATCC 700293T. Recently, ATCC suggested that there is a discrepancy between the previous report and the results obtained by ATCC in biochemical tests on T. medium ATCC 700293T. In this study, we re-examined and verified the biochemical characteristics of T. medium. The fermentation pattern of carbohydrates of T. medium resembled that of Treponema vincentii and Treponema denticola, but T. medium was clearly differentiated from T. vincentii in the production of indole, and from T. denticola in the hydrolysis of esculin. Also, sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) protein profile analysis and phylogenetic comparison of 16S rDNA sequences revealed that T. medium is clearly differentiated from any established treponemal species, which supports the validity of the proposal of Treponema medium as a new species.

Bacterial Proteins↗

IGF-I gene transfer by electroporation promotes regeneration in a muscle injury model.

The goal of this study was to determine whether insulin-like growth factor-I (IGF-I) gene delivery by electroporation promotes repair after muscle injury. An injury-repair model was created using mice in which a hamstring muscle was cut and sutured. A total of 50 microg of IGF-I DNA or green fluorescent protein (GFP) DNA (both in pCAGGS) was injected into the lesion and introduced into muscle cells by electrostimulation using an electric pulse generator. The number of regenerating muscle fibers in the IGF-I DNA group was significantly more than that in the GFP DNA group at 2 weeks after injection. The diameter of regenerating muscle fibers from the IGF-I DNA group was larger than that of the GFP DNA group at 4 weeks after injection. There was no significant difference in the serum IGF-I concentration between the IGF-I DNA group and the GFP DNA group at 1, 2, and 4 weeks after injection. However, muscle IGF-I concentration in the IGF-I DNA injection group was significantly greater than that in the GFP DNA injection group at 2 weeks after injection. These results demonstrated that the effects of enhanced IGF-I production were local and limited to the injected area. The ratio (injected/uninjected; intact) of the amplitude of compound muscle action potentials (CMAP) in the IGF-I DNA injection group was greater than that in the GFP DNA injection group at 4 weeks after injection and of the control group. In conclusion, IGF-I gene transfer by electroporation proved to be a simple, safe, inexpensive, and effective method to promote the regeneration of injured muscles in our injury model.

Animals↗

Paradoxical enhancement of spinal-cord-evoked potentials rostral and caudal to the site of progressive cord compression in the cat.

STUDY DESIGN: Analysis of the sequential waveform changes of the spinal-cord-evoked potentials (SCEPs) associated with progressive cord compression in the cat. OBJECTIVES: To document the phenomenon of paradoxical enhancement of SCEPs despite conduction abnormalities and to evaluate its possible significance. SETTING: Kochi Medical School, Kochi, Japan. METHODS: SCEPs were recorded simultaneously at four serial intervertebral levels, from T6-7 to T9-10 caudal to, and at three serial levels from T2-3 to T4-5 rostral to the compression site at T5-6 following epidural stimulation at L6 in 14 cats. RESULTS: Caudal to the compression site, the area of negative peak significantly increased toward maximal values of 277+/-36 (mean+/-SE), 151+/-9 and 110+/-4% as compared to the baseline precompression values (100%) at T6-7, T7-8, and T8-9, respectively. Rostral to the compression site, the area of negative peak significantly increased before subsequent deterioration and reached 105+/-2, 106+/-2, and 104+/-2% at T4-5, T3-4, and T2-3, respectively. The onset of negative peak enhancement, recorded either caudal or rostral to the compression site, showed a close temporal correlation (r>0.8, P&<0.001) with that of the prolongation in latency of SCEPs at T2-3. CONCLUSIONS: A progressive focal conduction block induced by compression of the spinal cord can paradoxically enhance the ascending SCEPs both caudally and, though less consistently, rostrally, representing a warning of the impending risk of paraplegia.

Animals↗

p53 Mutation and MDM2 amplification in inflammatory myofibroblastic tumours.

AIMS: The pathogenic mechanism and predictive indicators of biological behaviour of inflammatory myofibroblastic tumour are poorly understood. We investigated molecular abnormalities of p53 and MDM2 in order to assess whether these play an important role in pathogenesis, and whether they also contribute to clinicopathological aggressive phenotype in inflammatory myofibroblastic tumour. METHODS AND RESULTS: We compared the immunohistochemical expression of calponin, h-caldesmon, ALK, and p53 gene mutation and MDM2 gene amplification with clinicopathological findings in 15 cases of inflammatory myofibroblastic tumour. Histologically, cellular atypia was observed in five (33.3%) out of 15 cases. Local recurrences were observed in two (14.3%) of 14 informative cases, but no distant metastasis was observed. The expression of calponin (9/14; 64%) but not h-caldesmon (0/14; 0%) was seen, which suggested myofibroblastic differentiation. ALK expression was seen in eight (53.3%) out of 15 cases, particularly in patients under 40 years old. Nuclear expression of p53 protein was recognized in only one (6.7%) of 15 cases, and polymerase chain reaction single-strand conformation polymorphism followed by direct sequencing revealed p53 gene missense mutations in two (13.3%) of 15 cases. Nuclear expression of MDM2 was seen in four (26.7%) of 15 cases, and the MDM2 gene amplification was observed in two of the four cases. CONCLUSION: Inflammatory myofibroblastic tumour shows a wide spectrum of cellular atypia and biological behaviour with p53 and MDM2 expression. However, the alterations in the p53 pathway seem not to play a major role in the pathogenesis of inflammatory myofibroblastic tumour.

Adult↗

Actions of domperidone on the opioid system in isolated guinea-pig ileum: differences between dopamine antagonists on opioid system.

1. To validate the relationship between the dopaminergic, opioidergic and cholinergic nervous systems, we evaluated the effect of domperidone, a dopamine (D2) antagonist, on the opioid system in myenteric plexus-longitudinal muscle preparation isolated from the guinea-pig ileum. 2. One micromolar of domperidone did not affect the 0.1 Hz-evoked (duration 0.5 ms, maximum intensity) twitch response, but concentration dependently inhibited the twitch response between concentrations of 2 and 20 microM, and the inhibition was maximum after 20-30 min at the highest concentration used (20 microM). 3. Acetylcholine-evoked contraction on basal tension was also not inhibited by 1 microM domperidone, but the contraction was concentration dependently inhibited at concentrations of 10-100 microM in a non-competitive manner. 4. One micromolar of domperidone, however, increased post-tetanic twitch inhibition, an indicator of the release of endogenous opioids. This increase was completely antagonized by 1 microM naloxone. Twitch inhibition induced by dynorphin 1-13 (0.1-10 nM) was not affected by 1 microM domperidone, but increased the maximum twitch inhibition caused by morphine (0.1-1 microM). 6. These results might reflect the existence of an interaction between the dopaminergic and opioidergic system without the inhibition of the cholinergic system. Dopamine antagonists increased opioid action, an action that may depend more on the increased release of endogenous opioids than on supersensitivity of the opioid receptor.

Animals↗

Metastatic hepatocellular carcinoma of the maxillary sinus: a rare autopsy case without lung metastasis and a review.

A rare case of metastatic hepatocellular carcinoma (HCC) of the maxillary sinus in a 67-year-old man is reported along with the findings at autopsy. A fine needle aspiration biopsy specimen revealed a characteristic tumour structure resembling primary HCC. At autopsy, metastatic lesions were recognized in the bilateral adrenals, spleen, sternum, vertebrae and lymph node at the lesser curvature of the stomach, but not in the lung, suggesting that the HCC had metastasized to the maxillary sinus via the plexus venous vertebralis and/or the azygos vein, or lymph duct. In our reviewed 29 cases of metastatic HCC in the oro-maxillofacial region, most patients were men in the 50- to 70-year age range. At least 11 cases did not have lung metastasis, and in 18, metastasis to the oro-maxillofacial region was the first sign of HCC. One should be aware of the possibility to encounter the oral lesion as first sign of metastatic HCC.

Adrenal Gland Neoplasms↗