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Biomedical subjects

Y Ohta

Publications and source records attributed to Y Ohta.

At least 19 recordsLinked to original sources

Long-term treatment with TGFbeta1 impairs mechanotransduction in bovine aortic endothelial cells.

BACKGROUND AND PURPOSE: Vascular endothelial cells play a role in the physiological response to mechanical stress. Transforming growth factor beta1 (TGFbeta1) induces morphological changes in endothelial cells, and this may alter their mechanosensitive responses. The aim of this study was to examine the effects of TGFbeta1 on hypotonic stress (HTS)-induced responses in bovine aortic endothelial cells (BAECs). EXPERIMENTAL APPROACH: Cultured BAECs were treated with 3 ng ml(-1) TGFbeta1 for 24 h (24h-TGFbeta1) or 7 days (7d-TGFbeta1). Cytosolic actin fibres were stained with rhodamine-phalloidin. Intracellular Ca2+ concentration was measured using fura2. Tyrosine phosphorylation and RhoA expression were assessed by Western blotting. Expression of RhoA mRNA was assessed by real-time PCR. KEY RESULTS: BAECs developed pseudopod-like processes within 24 h and showed a fibroblast-like appearance after 7 days. HTS induced Ca2+ transients via endogenous ATP release in both control and 24h-TGFbeta1 BAECs but not in 7d-TGFbeta1 BAECs. We have previously shown that HTS-induced ATP release is mediated by sequential activation of RhoA and tyrosine kinases. The basal amount of membrane-bound RhoA was significantly lower in 7d-TGFbeta1 than in 24h-TGFbeta1 or control BAECs. HTS increased the membrane-bound RhoA to the same fractional level in 24h-TGFbeta1 and control BAECs, but its net maximal amount was significantly lower in 7d-TGFbeta1. HTS-induced downstream signals of RhoA activation, i.e. the tyrosine phosphorylation of FAK and paxillin, were markedly suppressed in 7d-TGFbeta1 BAECs. CONCLUSIONS AND IMPLICATIONS: These results indicate that long-term treatment with TGFbeta1 does not impair mechanoreception in BAECs but impairs mechanotransduction by affecting RhoA membrane translocation.

Actins↗

Sustained contraction and loss of NO production in TGFbeta1-treated endothelial cells.

BACKGROUND AND PURPOSE: Transforming growth factor beta1 (TGFbeta1) is generated in atherosclerotic and injured vessel walls. We examined whether the endothelial-to-mesenchymal transdifferentiation induced by TGFbeta1 affects endothelial functions. EXPERIMENTAL APPROACH: Bovine aortic endothelial cells (BAECs) were treated with 3 ng ml(-1) TGFbeta1 for 7 days. Contraction of TGFbeta1-treated BAECs was assessed by collagen gel contraction assay. Protein expression and phosphorylation were assessed by Western blotting. Intracellular Ca2+ concentration and NO production were measured using fura2 and DAF-2, respectively. KEY RESULTS: TGFbeta1-treated BAECs showed dense actin fibers and expressed smooth muscle marker proteins; they also changed into smooth muscle-like, spindle-shaped cells in collagen gel cultures. ATP (10 microM) induced a gradual contraction of collagen gels containing TGFbeta1-treated BAECs but not of gels containing control BAECs. ATP-induced contraction of TGFbeta1-treated BAECs was not reversed by the removal of ATP but was partially suppressed by a high concentration of sodium nitroprusside (1 microM). TGFbeta1-treated BAECs showed sustained phosphorylation of myosin light chain in response to ATP and low levels of basal MYPT1 expression. ATP-induced Ca2+ transients as well as eNOS protein expression were not affected by TGFbeta1 in BAECs. However, ATP-induced NO production was significantly reduced in TGFbeta1-treated BAECs. Anti-TGFbeta1 antibody abolished all of these TGFbeta1-induced changes in BAECs. CONCLUSIONS AND IMPLICATIONS: Mesenchymal transdifferentiation induced by TGFbeta1 leads to sustained contraction and reduced NO production in endothelial cells. Such effects, therefore, would not be beneficial for vascular integrity.

Animals↗

Isolation and characterization of a novel Burkholderia cepacia with strong antifungal activity against Rhizoctonia solani.

Strain CF-66 with strong antifungal activity against Rhizoctonia solani was isolated from compost samples. It is clearly demonstrated that strain CF-66 is belonging to Burkholderia cepacia complex by the morphological and biochemical tests and 16S rDNA sequence. The B. cepacia complex consists of a group of bacteria currently organized into nine genomovars, among them genomovar II and genomovar III, contain the highly epidemic strains. However, it was known that strain CF-66 is not a member of genomovar II or III of the B. cepacia complex by species-specific polymerase chain reaction assay. In this study, the antifungal compound CF66I produced by strain CF-66 was purified and characterized. Based on the nuclear magnetic resonance, GC-MS spectral and infrared spectral data, CF66I was confirmed to have amide bonds, alpha-methyl fatty acid, bromine, and some structural units such as CH(2)CH(2)O. CF66I is stable to high temperature, proteolytic enzymes, and organic solvents. CF66I inhibit the growth of a variety of plant pathogenic fungi and pathogenic yeast, whereas bacterial cells are unaffected. CF66I mainly reduced hyphal extension rates in a dose-dependent manner and induced severe change in cell morphology that resulted in swelled and formed very short hyphae with multiple branches.

Amides↗

Thoracoscopic robot-assisted bronchoplasty.

BACKGROUND: In robotic surgery, the ideal position of the system, as well as the optimal working angles and the proper positioning of the thoraco ports position is very important. No robot-assisted bronchoplasty has been reported. Our study describes use of the da VinciTM surgical system (Intuitive Surgical, Inc.) for robotic sleeve upper lobectomy in a human fresh cadaver. METHODS: A male cadaver was placed in the left lateral decubitus position. After thoracoscopic upper lobectomy was performed through the working port and the two ports, the robotic system was then set up behind the cadaver. The working port allowed introduction of the optical scope and the robotic surgical arms were inserted into the thoraco ports. The right bronchus was dissected and wedge was cut out with the robotic scissors. After standard lymph node dissection, end-to-end bronchial anastomosis was performed with robotic instruments. Once the anastomosis was complete, air leakage was checked with saline solution placed in the pleural cavity. RESULTS: Thoracoscopic robot-assisted bronchoplasty was performed successfully. CONCLUSIONS: In evaluating various positions of the system we demonstrated that our technique is sufficient approaches to robotic bronchoplasty. This procedure offers specific advantages over conventional bronchoplasty with accuracy and safety.

Anastomosis, Surgical↗

Ascorbic acid deficiency aggravates stress-induced gastric mucosal lesions in genetically scorbutic ODS rats.

We examined whether ascorbic acid (AA) deficiency aggravates water immersion restraint stress (WIRS)-induced gastric mucosal lesions in genetically scorbutic ODS rats. ODS rats received scorbutic diet with either distilled water containing AA (1 g/l) or distilled water for 2 weeks. AA-deficient rats had 12% of gastric mucosal AA content in AA-sufficient rats. AA-deficient rats showed more severe gastric mucosal lesions than AA-sufficient rats at 1, 3 or 6 h after the onset of WIRS, although AA-deficient rats had a slight decrease in gastric mucosal AA content, while AA-sufficient rats had a large decrease in that content. AA-deficient rats had more decreased gastric mucosal nonprotein SH and vitamin E contents and increased gastric mucosal lipid peroxide content than AA-sufficient rats at 1, 3 or 6 h of WIRS. These results indicate that AA deficiency aggravates WIRS-induced gastric mucosal lesions in ODS rats by enhancing oxidative damage in the gastric mucosa.

Animals↗

Management of malignant pleural effusion by multimodality treatment including the use of paclitaxel administered by 24-hour intrathoracic infusion for patients with carcinomatous pleuritis.

For successful intrapleural chemotherapy, intrapleural drug activity should be maintained for as long as possible. This interim report presents the results of treatment with paclitaxel administered by 24-hour intrathoracic infusion as an adjunct to selective surgical management and/or systemic chemotherapy for controlling malignant pleural effusion. Thirteen patients with carcinomatous pleuritis were enrolled in the study between October 2001 and September 2004. The sites of primary disease were the lung in 12 patients and the breast in one patient. Paclitaxel (120 mg/m2) was administered by 24-hour intrathoracic infusion. Seven patients underwent elective surgical treatment and 11 patients received adjuvant systemic chemotherapy. Mild toxicity occurred in 7 cases, and chest pain and neutropenia were dominant. During a median follow-up period of 9 months (range, 2-33 months), malignant effusion was controlled successfully in 11 patients (84.6%). The multimodality treatment, including the use of paclitaxel, in this manner merits further investigation for possible intervention for malignant pleural effusion originating in lung and breast neoplasms.

Adult↗

Evaluation of carcinomatous pleuritis on initial relapse in patients with lung cancer who underwent complete resection.

The purpose of this study was to evaluate the clinical characteristics and results of lung cancer patients with carcinomatous pleuritis on initial relapse. Over the past decade, seven patients out of 595 surgically treated lung cancer patients developed carcinomatous pleuritis as the initial sign of relapse. All patients had undergone complete resection of the primary peripheral type adenocarcinoma. Only 3 cases showed obvious visceral pleural invasion, while venous or lymphatic invasion were frequently found in the histological sections. The mean disease-free interval after surgery was 30.9 months. The median survival after the development of carcinomatous pleuritis was 14 months. The results showed the possible association of blood/lymphatic routes with the development of this type of recurrence. In some lung cancer patients with carcinomatous pleuritis on initial relapse, long-term survival can be expected by aggressive interventions consisting of intrathoracic and/or systemic chemotherapies.

Adenocarcinoma↗

Reduction of a vascular endothelial growth factor receptor, fetal liver kinase-1, by antisense oligonucleotides induces motor neuron death in rat spinal cord exposed to hypoxia.

Vascular endothelial growth factor (VEGF) is reported to play a neuroprotective role through a VEGF receptor, fetal liver kinase-1 (Flk-1) in vitro. We investigated whether reduction of Flk-1 could induce motor neuron loss in rat spinal cord by inhibiting the expression of Flk-1 in rat spinal cord using antisense oligodeoxynucleotides (ODNs) against the Flk-1 receptor. Rat spinal cord was repetitively exposed to 12% hypoxia, and the change of the phosphatidylinositol 3-kinase (PI3-K)/Akt pathway and the mitogen-activated protein kinase kinase (MEK)/extracellular-signal-regulated kinase (ERK) pathway was examined. Intrathecal infusion of Flk-1 antisense ODNs for 7 days suppressed almost completely Flk-1 expression in the lumbar segment of the spinal cord and was followed by a hypoxic challenge with 12% oxygen for 1 h that was repeated for 7 more days. In the lumbar segment, we observed that reduced Flk-1 expression and hypoxic challenge for 7 days resulted in approximately 50% loss of motor neurons, in which the activation of Akt and ERK, that is, increased levels of phosphorylated-Akt and of phosphorylated-ERK by hypoxia, was markedly inhibited. In contrast, the reduction of Flk-1 expression alone did not induce motor neuron loss. These results suggest that VEGF exerts its protective effect on motor neurons against hypoxia-induced toxicity by the Flk-1 receptor through the PI3-K/Akt and the MEK/ERK signaling pathways.

Animals↗

[Surgical results of non-small cell lung cancer invading parietal pleura and chest wall].

This retrospective analysis was undertaken to review our results of treatment of lung cancers with invasion of non-apical and non-vertebral chest wall structures. In summary of our experience, although relatively good prognosis can be expected in N0M0 patients with the histological type of adenocarcinoma by initial operation, distant relapse remains a major problem of the disease. Furthermore, our results are in agreement with the idea that postoperative adjuvant therapy is of little value in patients with complete resection. To ameliorate surgical outcomes, induction treatment should be considered and preoperative staging assessment needs to be strictly done for proper selection of patients with this locally advanced disease. The indication of initial operation needs to be cautiously determined for patients with this disease.

Carcinoma, Non-Small-Cell Lung↗

Hyper-production of an isomalto-dextranase of an Arthrobacter sp. by a proteases-deficient Bacillus subtilis: sequencing, properties, and crystallization of the recombinant enzyme.

Arthrobacter globiformis T6 is unique in that it produces an enzyme yielding only isomaltose from dextran. In the present study, the organism was re-identified and its classification as a new species of the genus Arthrobacter, A. dextranlyticum, was proposed. The high G+C gene (66.8 mol%) for the isomalto-dextranase was sequenced. The deduced amino acid sequence, with a calculated molecular mass of 65,993 Da (603 amino acids), was confirmed by nanoscale capillary liquid chromatography coupled to tandem mass spectrometry, which covered 71.1% of the amino acid residues of the entire sequence. The enzyme was grouped into glycoside hydrolase family 27, and the C-terminal domain has homology to carbohydrate-binding module family 6. Hyper-exoproduction of the recombinant enzyme was achieved at a level corresponding to approximately 4.6 g l(-1) of culture broth when proteases-deficient Bacillus subtilis cells were used as the host. The purified enzyme (65.5 kDa) had an optimal pH and temperature for activity of 3.5 and 60 degrees C, respectively. It was crystallized using the sitting-drop vapor-diffusion method at 293 K.

Amino Acid Sequence↗

Enzymatic properties and nucleotide and amino acid sequences of a thermostable beta-agarase from a novel species of deep-sea Microbulbifer.

An agar-degrading bacterium, strain JAMB-A7, was isolated from the sediment in Sagami Bay, Japan, at a depth of 1,174 m and identified as a novel species of the genus Microbulbifer. The gene for a novel beta-agarase from the isolate was cloned and sequenced. It encodes a protein of 441 amino acids with a calculated molecular mass of 48,989 Da. The deduced amino acid sequence showed similarity to those of known beta-agarases in glycoside hydrolase family 16, with only 34-55% identity. A sequence similar to a carbohydrate-binding module was found in the C-terminal region of the enzyme. The recombinant agarase was hyper-produced extracellularly using Bacillus subtilis as the host, and the enzyme purified to homogeneity had a specific activity of 398 U (mg protein)(-1) at pH 7.0 and 50 degrees C. It was thermostable, with a half-life of 502 min at 50 degrees C. The optimal pH and temperature for activity were around 7 and 50 degrees C, respectively. The pattern of agarose hydrolysis showed that the enzyme was an endo-type beta-agarase, and the final main product was neoagarotetraose. The activity was not inhibited by NaCl, EDTA, and various surfactants at high concentrations. In particular, sodium dodecyl sulfate had no inhibitory effect up to 2%.

Alteromonadaceae↗

The results of therapy for bilateral multiple primary lung cancers: 30 years experience in a single centre.

AIMS: This study reviews our 30 years experience in the clinical assessment and surgical management of bilateral multiple primary lung cancer (BMPLC). METHODS: Between January 1973 and December 2001, 1906 patients with primary lung cancer underwent surgical resection in Kanazawa University Hospital. Thirty-seven patients (1.9%) who had developed a BMPLC using the criteria of Martini and Antakli. RESULTS: Eighteen patients had synchronous lesions, and 18 patients had metachronous lesions. One patient had synchronous and metachronous lesions. Overall 10-year survival was 56%. The actuarial 5-year survival for bilateral synchronous cancers was 69%, median survival (MST) 90 months (range 8-153 months), and 10-year survival was 47%. The actuarial 5-year survival for second metachronous cancers was 51%, with an MST of 114 months (range 6-192 months). CONCLUSION: Aggressive surgical therapy is effective in patients with a bilateral MPLC if they satisfy the usual criteria of operability. The surgical methods that preserve healthy lung tissue such as sleeve resection and limited resection must be selected in compliance with cancer characters.

Adult↗

Improvement of blood pressure control in a hypertension clinic: a 10-year follow-up study.

The objective of the study was to assess whether the publication of new guidelines, such as JNC VI 1997 and WHO/ISH 1999, and the development of new antihypertensive drugs have improved blood pressure (BP) control. A total of 150 patients (age 29-88, mean 66+/-11 years in 2001) who were followed at our hypertension clinic during 1991-2001 were retrospectively investigated. We compared the clinical characteristics of the patients in 2001 to those in 1991 and 1996, using the averaged BP determined at two occasions each year for our analysis. The average BP decreased during the 10 years between 1991 and 2001. When good BP control was defined as <140/90 mmHg, the rate of patients with good BP control increased from 31% in 1991 to 43% in 1996, and to 57% in 2001 (P<0.001 vs 1991). Both younger (< or =64 years) and older (> or =65 years) patients showed similar improvement during these 10 years. In 2001, satisfactory BP control (<130/85 mmHg) was achieved in 24% of younger patients, which was significantly higher than the achievement in 1991 (10%, P=0.02). This improvement occurred at the same time as an increase in the prescription of Ca antagonists and angiotensin II antagonist. The patients with improved BP control during these 10 years (n=50) showed lower body mass index (BMI) and serum total cholesterol levels in 2001 compared to persistently uncontrolled patients (n=54). Furthermore, the change in BMI during these 10 years was significantly less in the patients with improved BP control than in the persistently uncontrolled patients. In conclusion, BP control improved in the 10 years studied, and it seems to be attributable to the more frequent use of the newer drugs such as angiotensin II antagonists and Ca antagonists, to lifestyle modification and also to the growth in awareness of the importance of strict BP control.

Adult↗

Comparison between broilers and layers for growth and protein use by embryos.

Three experiments were conducted to compare the growth and protein utilization of embryo between broilers and layers. In experiments 1 and 2, the average weight of eggs was the same for broilers and layers. Nothing or an amino acid (AA) solution was injected into the eggs of broilers at d 7 of incubation, and the plasma AA concentration of newly hatched chicks was determined in broilers in experiment 1. In experiment 2, the same treatments as experiment 1 were used on layer breeder eggs. Plasma Tau, Thr, and Lys concentrations of hatched chicks increased when AA solution was injected in broilers breeder eggs (P < 0.05) but not in layers (P > 0.05). The AA ratio to Lys was reduced by AA injection in broilers but not in layers. In experiment 3, weights of embryos and egg were recorded, and CP contents were analyzed over time during incubation (d 0, 7, 14, and 19 of incubation) in broilers and layers using eggs of the same weight. There were no differences in the weights and CP contents of embryos and eggs from broilers and layers. On d 14 and 19 of incubation, weights and CP contents of embryo were higher in broilers than layers (P < 0.05). These results suggested that the egg protein content might be adequate for hatching but insufficient for maximum growth of embryos from broilers.

Amino Acids↗

Role of foxj1 and estrogen receptor alpha in ciliated epithelial cell differentiation of the neonatal oviduct.

Estrogen regulates proliferation and differentiation of epithelial cells in the mammalian oviduct, but pathways for cell-specific differentiation are not well understood. In the epithelial cells of the developing rat oviduct, we found estrogen receptor (ER) alpha is expressed at birth and persists in all cells through neonatal day (ND) 7 when ciliated cells appear. To determine a specific function of ER and foxj1, a transcription factor known to have fundamental roles in ciliogenesis in the lung, in differentiation of the ciliated epithelial cells, we treated newborn rats from ND 0 to 5 with estradiol-17beta (E2) with and without a selective ER antagonist. E2 enhanced the number of proliferating cells and accelerated the process of epithelial cell differentiation resulting in ciliogenesis by ND 5, and co-treatment with an ER antagonist inhibited these changes. Foxj1 was expressed only in the infundibulum and ampulla (INF/AMP). That expression preceded the appearance of cilia and was induced by E2. Cilia were absent in oviducts of foxj1-deficient mice, indicating that foxj1 plays a critical role in oviductal ciliogenesis. However, we found the presence of cilia in the ERalpha-deficient mouse oviduct. The widespread expression of ERalpha in oviductal epithelium, but restriction of cilia to the INF/AMP regions, and importantly, the presence of cilia in the ERalpha-deficient mice, suggested ER signaling is not essential for ciliated epithelial cell differentiation. These observations demonstrate that, although E2 stimulates the differentiation process of ciliated epithelial cells, foxj1 is directly required for epithelial cell ciliogenesis of the neonatal oviduct.

Animals↗

[Perioperative targeting brachytherapy for lung cancer invading the chest wall].

We evaluated the efficacy of perioperative targeting brachytherapy for lung cancer invading the chest wall. Between 1998 and 2003, 7 patients underwent perioperative targeting brachytherapy for lung cancer invading the chest wall. There were 5 male and 2 female patients. The mean age was 63.3 years, with a range of 45 to 77 years. All patients underwent complete resection including the chest wall combined resection. During the operation, plastic afterloading catheters fixed on the Vicryl mesh at interval of 1 cm were placed on the site of chest wall resection. From the third to sixth day after the operation, 15 to 32 Gy of radiation was delivered over 3 or 4 days using a high dose rate remote afterloading system. The area targeted for brachytherapy was determined by a computed tomography (CT) scanner translator with a computer program for radiation planning. The median postoperative hospital stay was 35 days. Local recurrences were observed in 2 patients, but there was no evidence of recurrence in the margin of the resected chest wall. We believe that this short period of treatment and the low side effects enhances the quality of the patients. Prevention of local recurrence was achieved in short term follow-up.

Aged↗

Hand-assisted laparoscopic resection of serous cystadenoma of the pancreas.

It is difficult to exclude the possibility of malignancy of pancreatic cystic tumors because a biopsy of the pancreas is hard to obtain. The indication of open surgery for those cystic tumors without evidence of malignancy is controversial. Therefore, laparoscopic or laparoscopically assisted procedure would be an adequate choice of treatment for cystic tumors of the pancreas. Hand-assisted laparoscopic distal pancreatectomy with preservation of the spleen and the splenic artery and vein was performed for two cases of pancreatic cystic tumors. Three ports and one hand port were used. After careful dissection and accurate hemostasis between the pancreas and splenic vessels, laparoscopic distal pancreatectomy was carried out using an endoscopic linear stapler. There were no perioperative complications. The pathological diagnoses were oligocystic serous cystadenoma and solitary cystic serous cystadenoma, respectively. Hand-assisted, spleen-preserving laparoscopic distal pancreatectomy with preservation of the splenic artery and vein is a feasible procedure for the treatment of benign or borderline-malignant cystic lesions of the distal pancreas.

Adult↗