Gene list on a plant tumor-inducing plasmid, pti-SAKURA in Agrobacterium tumefaciens MAFF301001.
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Biomedical subjects
Publications and source records attributed to K Iwata.
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We have previously demonstrated that chemically oversulfated fucoidan (OSF) but not native fucoidan (NF) effectively suppresses the tube structure formation by human umbilical vein endothelial cells (HUVEC) on the basement membrane preparation, Matrigel. In this study, using more defined systems where basic fibroblast growth factor (bFGF) induces the tube formation by HUVEC on collagen gel, we investigated the mechanism responsible for the inhibition of angiogenesis by OSF in vitro. Unlike NF and desulfated fucoidan (desF), OSF potently inhibited the bFGF-induced HUVEC migration and tube formation. ELISA for tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) in the culture media indicated that OSF increased the bFGF-induced release of PAI-1 antigen, but not of t-PA antigen. Analyses of the binding of bFGF to HUVEC surfaces and the following protein tyrosine phosphorylation revealed that OSF could promote the cell binding and autophosphorylation of 140 and 160 kDa receptors. In heparitinase-treated HUVEC, contrarily, the bFGF binding and PAI-1 release were decreased by OSF. These results suggest that OSF is a highly sulfated unique polysaccharide that can promote the binding of bFGF to the heparan sulfate molecules required for binding to the high affinity receptors with tyrosine kinase activity. The resultant increase in PAI-1 release may play a key role for the prevention of cell migration accompanied by matrix proteolysis.
To investigate the effect of metformin on intestinal glucose absorption, a perfusion study of the intestine was performed in the rat. Male Wistar albino rats (8 weeks old) were used in the present study. The glucose absorption by the perfused intestine (788.1+/-81.9 micromol/30 min) was not changed significantly by the direct addition of metformin (90 microg/mL) to the perfusing medium (737.0+/-118.2 micromol/30 min) or by intraduodenal metformin (250 mg/kg in saline solution) infusion (772.8+/-106.3 micromol/30 min). In rats orally administered metformin (250 mg/kg) for 5 days, glucose absorption by the perfused intestine (375.0+/-164.3 micromol/30 min) was significantly (P<0.001) lower than that in control rats (811.0+/-83.1 micromol/30 min). These results indicate that metformin had a significant effect on the digestive tract, and that metformin treatment exerted an inhibitory effect on intestinal glucose absorption in the rat.
Metastasis is the spread of tumors culminating in the establishment of one or more secondary tumors at remote sites. In deciding the best treatment for cancer therapy, estimations of the colony size of metastatic tumors and predictions of the future spread of colonies are needed. A dynamical model for the colony size distribution of multiple metastatic tumors is presented here. The dynamics is described by equations that incorporate both the colonization by metastasis and the growth of each colony. When the colony growth is subject to the Gompertz function, the explicit solution obtained tends to an asymptotic stable distribution that shows a monotonically decreasing or U-shaped pattern according to the values of clinically significant parameters, such as the colonization coefficient and the fractal dimension of blood vessels. This predicted colony size distribution agrees well with successive data of a clinically observed size distribution of multiple metastatic tumors of liver cancer. The combined analysis of the theoretical colony size distribution and clinical data will give useful information on the diagnosis and the therapy for cancer patients.
Crown gall tumor disease in dicot plants is caused by Agrobacterium tumefaciens harboring a giant tumor-inducing (Ti) plasmid. Here, for the first time among agrobacterial plasmids, the nucleotide sequence of a typical nopaline-type Ti plasmid (pTi-SAKURA) was determined completely. In total, 195 open reading frames (ORFs) were estimated in the 206479 bp long sequence. 20 genes for conjugation, three for replication, 22 for pathogenesis and 37 for genetic colonization of host plants were found within two-thirds of the plasmid. These genes formed seven functional gene clusters with narrow inter-cluster spaces. In the remaining one-third of the plasmid, novel genes including homologs of mutT, Rhizobium nodQ and Sphingomonas ligE genes were found, which are likely to be responsible for the broad host range. Restriction fragment length variation indicates extreme plasticity of the part required for conjugational gene transfer and the above-mentioned one-third of the plasmid, even among closely related Ti plasmids.
Chiral discrimination of racemic carbazole carbonyl (CC)-amino acids with linear alkyl sidechain (C(1)-C(4)) by bovine serum albumin (BSA) was investigated by competitive replacement experiments using dansyl-L-proline and dansyl-D-norvaline as fluorescent probes. It was found that the CC derivatives of the D-forms of alanine (C(1)), amino butyric acid (C(2)), norvaline (C(3)), and norleucine (C(4)) bound to the dansyl-L-proline site much more strongly than their L-forms, whereas the interactions between both enantiomers of these amino acids with dansyl-D-norvaline site were slight.
The self-assembly of the soluble peptide Abeta into Alzheimer's disease amyloid is believed to involve a conformational change. Hence the solution conformation of Abeta is of significant interest. In contrast to studies in other solvents, in water Abeta is collapsed into a compact series of loops, strands, and turns and has no alpha-helical or beta-sheet structure. Conformational stabilization is primarily attributed to van der Waals and electrostatic forces. A large conspicuous uninterrupted hydrophobic patch covers approximately 25% of the surface. The compact coil structure appears meta-stable, and because fibrillization leads to formation of intermolecular beta-sheet secondary structure, a global conformational rearrangement is highly likely. A molecular hypothesis for amyloidosis includes at least two primary driving forces, changes in solvation thermodynamics during formation of amyloid deposits and relief of internal conformational stress within the soluble precursor during formation of lower-energy amyloid fibrils.
Collagen synthesis in vascular smooth muscle cells (SMCs) after exposure to tumor necrosis factor alpha (TNF-alpha) was investigated using a culture system. The synthesis of collagenase-digestible proteins (CDP) and noncollagenous proteins (NCP) was evaluated by the [3H]proline incorporation. It was shown that TNF-alpha markedly suppresses the incorporation of [3H]proline into both CDP and NCP in confluent cultures of SMCs but not in sparse cultures of the cells. Such a marked suppression by TNF-alpha was not observed in confluent bovine aortic endothelial cells and human fibroblastic IMR-90 cells. In confluent SMCs, the synthesis of CDP was more strongly inhibited by TNF-alpha than that of NCP. When the CDP synthesis was stimulated by transforming growth factor beta, TNF-alpha suppressed the stimulation in both confluent and sparse SMCs. Human SMCs synthesized types I, III, IV and V collagens; TNF-alpha markedly decreased the relative proportion of types IV and V. It was therefore suggested that TNF-alpha modulates the collagen synthesis by SMCs depending on their cell density and modifies the formation of atherosclerotic lesions.
A total of 68 neurons were recorded from the ventro-postero-lateral nucleus of thalamus (VPL) in rats with a unilateral chronic constriction injury (CCI) of the sciatic nerve (n=20), sham operation (n=24) and naive rats (n=24), and effects of the lesion of dorsal column (DC) pathway [DC lesion or DC+gracile nucleus lesions] on VPL nucleus neuronal activities were studied. In the VPL nucleus contralateral to the CCI (receiving input from the injured nerve), response latencies of low threshold mechanoreceptive (LTM) and wide dynamic range (WDR) neurons to electrical stimulation of the sciatic nerve were significantly longer than that in the contralateral VPL nucleus receiving input from the sham-operated side (P<0.05). In contrast, response latencies of LTM and WDR neurons to DC stimulation were not different between the sham operated and CCI sides (0.05). Background activity of WDR neurons was significantly higher in the VPL nucleus contralateral to the CCI side when compared to neurons in the VPL nucleus contralateral to the sham operated side and in naive animals. Responses of LTM and WDR neurons to innocuous mechanical stimulation of the receptive fields were significantly decreased after DC and DC+gracile nucleus lesions in all animals. However, the responses of WDR neurons to noxious stimuli were selectively reduced only in rats with CCI by DC and DC+gracile nucleus lesions (P<0.05). The decrease in noxious stimulus-evoked responses of WDR neurons in the VPL nucleus contralateral to the CCI side after DC and DC+gracile nucleus lesions was greater than that in the VPL nucleus contralateral to the sham operated side and naive animals. These results indicated that DC and DC+gracile nucleus lesions produced selective and stronger effect on noxious responses of VPL nucleus WDR neurons receiving input from the site of nerve injury. The findings suggest that the gracile nucleus-thalamic pathway conveys, or modulates, nociceptive information to the VPL nucleus following peripheral nerve injury, resulting in an increase in VPL nucleus response to noxious stimuli that contributes to the development of mechanical hyperalgesia.
A one-step sandwich enzyme immunoassay (EIA) system for human matrix metalloproteinase 20 (MMP-20, enamelysin) was established by use of a solid-phase monoclonal antibody and a separate peroxidase-labeled monoclonal antibody. The EIA system was shown to be sensitive and quantitative for the detection of MMP-20. As little as 1.0 ng/ml (50 pg/assay) of MMP-20 protein could be reliably detected. The EIA system was linear over a range of 2.5-160 ng/ml (125-8,000 pg/assay), and the EIA system was versatile in that it was capable of detecting with equal sensitivity proMMP-20, active MMP-20, and MMP-20 with COOH-terminal deletions. The EIA system was validated by the successful detection of MMP-20 in the culture medium of Chinese hamster ovary cells (CHO-K1) that were transfected with an MMP-20 expression vector. No MMP-20 was detected in normal human serum, normal saliva, or in selected tumors. However, when recombinant human MMP-20 was added to human saliva, the EIA system did detect quantifiable amounts of the MMP-20, indicating that the system will work within the framework of complex in biological fluids.
OBJECTIVE: To examine the effectiveness of Ninjin Yoei To (NYT; Ren-Shen-Yang-Rong-Tang in Chinese medicine; Kotaro Pharmaceutical Co., Ltd., Osaka, Japan), one of the traditional herbal medicines, against lung carcinoma. SETTING: The Nursing Center Himawari DESIGN, PATIENT, AND PREPARATION: The regular dosage of NYT (15 g/d) was prescribed for 7 weeks to one elderly patient with lung carcinoma. The daily standard dose of NYT is prepared from dried extract obtained from 12 crude natural substances, ginseng, cinnamon bark, Japanese angelica root, astragalus root, peony root, citrus unshiu peel, rehmannia root, polygala root, atractylodes rhizome, schisanda fruit, poria sclerotium, and glycyrrhiza. NYT is certified by the Japanese Ministry of Health and Welfare. RESULTS: The tumor marker levels (CEA and CA19-9) decreased and the scores of yin-yang and xu-shi inverted from negative and positive during 7 weeks. The patient's cough disappeared and her appetite recovered. CONCLUSION: NYT has a positive effect on life expectancy for patients with malignancy. The diagnostic scoring system in yin-yang and xu-shi and prescription of Chinese herb may be available to gain control over a patient's health.
The organization and response properties of nociceptive neurons in area 1 of the primary somatosensory cortex (SI) of anesthetized monkeys were examined. The receptive fields of nociceptive neurons were classified as either wide-dynamic-range (WDR) neurons that were preferentially responsive to noxious mechanical stimulation, or nociceptive specific (NS) that were responsive to only noxious stimuli. The cortical locations and the responses of the two classes of neurons were compared. An examination of the neuronal stimulus-response functions obtained during noxious thermal stimulation of the glabrous skin of the foot or the hand indicated that WDR neurons exhibited significantly greater sensitivity to noxious thermal stimuli than did NS neurons. The receptive fields of WDR neurons were significantly larger than the receptive fields of NS neurons. Nociceptive SI neurons were somatotopically organized. Nociceptive neurons with receptive fields on the foot were located more medial in area 1 of SI than those with receptive fields on the hand. In the foot representation, the recording sites of nociceptive neurons were near the boundary between areas 3b and 1, whereas in the hand area, there was a tendency for them to be located more caudal in area 1. The majority of nociceptive neurons were located in the middle layers (III and IV) of area 1. The fact that nociceptive neurons were not evenly distributed across the layers of area 1 suggested that columns of nociceptive neurons probably do not exist in the somatosensory cortex. In electrode tracks where nociceptive neurons were found, approximately half of all subsequently isolated neurons were also classified as nociceptive. Low-threshold mechanoreceptive (LTM) neurons were intermingled with nociceptive neurons. Both WDR and NS neurons were found in close proximity to one another. In instances where the receptive field shifted, subsequently isolated cells were also classified as nociceptive. These data suggest that nociceptive neurons in area 1 of SI are organized in vertically orientated aggregations or clusters in layers III and IV.
OBJECTIVES: The flap valve principle is not easily applicable to an ileal pouch since a submucosal tunnel is difficult to create. We attempted to construct an ileal pouch with an umbilical stoma applying the serous-lined principle for both ureteral implantation and construction of a continent valve. METHODS: In 9 patients, a continent pouch with an umbilical stoma was created entirely from an ileal segment. Adopting the serous-lined principle, a continent valve was created by appendix in 2 cases, tapered ileum in 3 cases, and reconfigured ileum in 4 cases. These were implanted into the anterior suture line of the pouch and embedded into the serous-lined tunnel formed by the pouch wall. RESULTS: Patients can catheterize the pouch easily with a 14-Fr catheter. Postoperatively, pouch capacity is over 400 ml with complete continence. CONCLUSION: This technique can provide a simple and effective continent ileal pouch facilitating umbilical anastomosis. As a continent valve, the reconfigured ileal segment seems most useful for application of the serous-lined principle.
To examine the transition to ureogenesis, the gobiid fish Mugilogobius abei was immersed in 2 mmol l(-)(1) NH(4)HCO(3) or a (15)N-labelled ammonia solution [1 mmol l(-)(1) ((15)NH(4))(2)SO(4), pH 8.0] for 4-8 days. When exposed to 2 mmol l(-)(1) NH(4)HCO(3) or (15)N-labelled ammonia solution for 4 days, the rate of urea excretion increased to seven times that of the control (in 20 % synthetic sea water) and remained at this level for 4 days. The proportion of nitrogen excreted as urea reached 62 % of total nitrogen excretion (ammonia-N + urea-N). (15)N-enrichment of the amide-N in glutamine in the tissues of fish exposed to (15)N-labelled ammonia was virtually the same as that of ammonia-N: i.e. approximately twice that of urea-N in the excreta and the tissues. Glutamine contents and glutamine synthetase activities in the liver and muscle increased greatly following exposure to ammonia. Urea and citrulline contents in the muscle and whole body of the exposed fish increased significantly, whereas uric acid contents remained unchanged. Carbamoyl phosphate synthetase III (CPSase III) mRNA expression and CPSase III activity were detected in the muscle, skin and gill, but levels were negligible in the liver. Furthermore, all other ornithine-urea cycle (O-UC) enzymes were also detected in muscle, skin and gill. Thus, M. abei clearly shows the transition from ammoniotely to ureotely under ammonia-loading condition and is able to produce urea mainly via the O-UC operating in multiple non-hepatic tissues as a means for ammonia detoxification.
The aim of this study was to examine whether a specific PPARgamma ligand can inhibit the growth of human pancreatic cancer cells through induction of terminal differentiation. PPARgamma was expressed in five human pancreatic cancer cell lines: Capan-1, AsPC-1, BxPC-3, PANC-1, and MIA PaCa-2. Treatment of these cells with a specific PPARgamma ligand, thiazolidinedione (TZD), resulted in inhibition of both cellular and clonogenic growth, and G1 cell cycle arrest. Finally, thiazolidinedione treatment resulted in induction of p21WAF-1 and increased expression of differentiation markers. These results suggest that thiazolidinedione treatment inhibits growth and induces cellular differentiation in pancreatic cancer cells and thereby reduces their development in favor of differentiated and stable cell phenotype.
Hepatitis C virus (HCV) infection induces a variety of extrahepatic manifestations such as oral lichen planus (OLP). To clarify the role of HCV in the development of OLP, we investigated the occurrence of OLP in patients with chronic hepatitis C treated with interferon (IFN). Of 275 patients with chronic hepatitis C, 6 developed OLP during the IFN treatment. However, OLP developed in none of 230 patients with chronic hepatitis C who did not undergo the IFN therapy. The IFN treatment in chronic hepatitis C patients developed OLP significantly, as compared with the non-treated group (p < 0.05). 4 of 6 patients who developed OLP during the IFN treatment had a complete response with normalization of ALT levels and undetectable HCV RNA after the treatment. There were no significant correlations between the effect of the IFN treatment and outcome of OLP. Furthermore, 3 of the 6 patients developed OLP, when serum HCV RNA became negative. These results suggest that direct viral factors may not be important in the pathogenesis of OLP in patients with chronic hepatitis C. Immunological changes caused by IFN may play a role in the development of OLP associated with HCV infection.
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We attempt to differentiate the physiological state during sensory deprivation (SD) from normal sleep and wakefulness in terms of electroencephalogram (EEG) and heart rate variability (HRV) dynamics. Kullback-Leibler (K-L) divergence is employed to quantify differences between their state-dependent dynamics. As a result, the dynamics of EEG and HRV during SD are found to be far distant from any representative dynamics of natural states of sleep and wakefulness. However, relatively speaking, the findings in SD can be categorized into two patterns. (a) The dynamics of HRV during SD is similar to that of rapid eye movement (REM) sleep, and the dynamics of EEG during SD is similar to that of wakefulness. (b) The dynamics of both HRV and EEG during SD are similar to that of REM. Such dissociation between states classified by EEG and HRV dynamics frequently takes place during SD. These findings suggest the peculiarity of the physiological state during the SD distinct from sleep and wakefulness.