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Biomedical subjects

T Fujiwara

Publications and source records attributed to T Fujiwara.

At least 37 records · Page 2Linked to original sources
PubMed2003

A novel method for gene delivery and expression in esophageal epithelium with fibrin glues containing replication-deficient adenovirus vector.

BACKGROUND: Gene transfer to the esophageal epithelium holds the potential for the therapy of malignant as well as premalignant lesions in the upper gastrointestinal tract. Replication-deficient recombinant adenoviruses represent an efficient means of introducing genes in vivo into cells in a variety of organs. The majority of in vivo studies utilize direct submucosal injection for delivery of the viral vectors into the locoregional area of the gut; transferring genes into epithelial cells, however, is difficult because viruses are retained in the subepithelial space. To establish the efficient method for gene transfer into the epithelial cells, we have developed a multiluminal spray catheter that can be passed through the accessory channel of an endoscope, and we have evaluated the feasibility of fibrin glues as a vehicle of recombinant adenoviruses in a porcine model. METHODS: The fibrinogen solution and the thrombion solution containing an E1/E3 deleted recombinant adenovirus expressing the bacterial lacZ gene (Ad-lacZ) were endoscopically sprayed on the porcine esophagus through the catheter attached to the dual-barrel syringe. Twenty-four hours after gene delivery, beta-galactosidase activity of the esophagus was determined under the microscope following X-gal staining. RESULTS: The fibrin glue could be locally sprayed on the porcine esophagus by using the multichannel catheter through the endoscope. Attachment of the fibrin glue comtining Ad-lacZ caused strong beta-galactosidase staining on epithelial cells in the mucosal surface, but not in the basal cell layer. CONCLUSION: Endoscopic local delivery of recombinant adenoviruses in aerosolized fibrin glues through a multiluminal catheter could provide an optimal technique for gene transfer into epithelial cells in the mucosal surfece, which may have important implications for the treatment of human esophageal premalignant diseases.

Adenoviruses, Human↗
PubMed2003

Nitric oxide synthase inhibition by N(G)-nitro-L-arginine methyl ester retards vascular sprouting in angiogenesis.

The present study was designed to evaluate the role of nitric oxide (NO) for angiogenesis. Angiogenesis was elicited upon mouse cornea by chemical cautery with silver nitrate. Angiogenic activity was evaluated by measuring the length of vascular sprout with or without administration of NO synthase (NOS) inhibitor, N((G))-nitro-L-arginine-methyl ester (L-NAME). In the pericorneal plexus, a circulatory loop situated in the same topological situation for all individuals was selected to observe vascular sprouting. At 72 h after cauterization, the length of the longest vascular sprout was measured using the perfused whole-mount cornea. The length of nontreated mice (83 +/- 83 microm) was significantly longer than that of L-NAME treated mice (33 +/- 24.6 microm). To address the possible contribution of production of vascular endothelial growth factor (VEGF) and NO, we measured mRNAs of VEGF and inducible NOS. The mRNA level of VEGF increased to 170% of the nontreated level at 12 h after cauterization and returned to the nontreated level by 24 h after cauterization. mRNA of inducible NOS remained elevated 24 h after cauterization. These results suggest that the response of preexisting vessels to angiogenic stimulus via NO is of importance in the process of angiogenesis, i.e., vascular sprouting is promoted by NO production. This might be attributable to enhancement of an increase in vascular permeability and /or vasodilation via NO.

Animals↗
PubMed2003

Simple and fast lateral flow test for classification of leprosy patients and identification of contacts with high risk of developing leprosy.

The interruption of leprosy transmission is one of the main challenges for leprosy control programs since no consistent evidence exists that transmission has been reduced after the introduction of multidrug therapy. Sources of infection are primarily people with high loads of bacteria with or without clinical signs of leprosy. The availability of a simple test system for the detection of antibodies to phenolic glycolipid-I (PGL-I) of Mycobacterium leprae to identify these individuals may be important in the prevention of transmission. We have developed a lateral flow assay, the ML Flow test, for the detection of antibodies to PGL-I which takes only 10 min to perform. An agreement of 91% was observed between enzyme-linked immunosorbent assay and our test; the agreement beyond chance (kappa value) was 0.77. We evaluated the use of whole blood by comparing 539 blood and serum samples from an area of high endemicity. The observed agreement was 85.9% (kappa = 0.70). Storage of the lateral flow test and the running buffer at 28 degrees C for up to 1 year did not influence the results of the assay. The sensitivity of the ML Flow test in correctly classifying MB patients was 97.4%. The specificity of the ML Flow test, based on the results of the control group, was 90.2%. The ML Flow test is a fast and easy-to-perform method for the detection of immunoglobulin M antibodies to PGL-I of M. leprae. It does not require any special equipment, and the highly stable reagents make the test robust and suitable for use in tropical countries.

Antibodies, Bacterial↗
PubMed2003

Expression of the exocytotic protein syntaxin in mouse oocytes.

Syntaxin is an integral membrane protein that is involved in membrane fusion. The exocytosis of the contents of cortical granules, secretory vesicles located in the cortex of an egg, modify the extracellular environment to block additional spermatozoa from penetrating the newly fertilized egg. The aim of this study was to characterize syntaxin expression in mouse oocytes, and to determine the specific isoform that is expressed. Syntaxin was demonstrated in the mouse ovary and in mouse oocytes by both western blot and reverse transcription-polymerase chain reaction analyses. Syntaxin 4 was specifically expressed in metaphase II oocytes. Syntaxin was also immunolocalized within metaphase II oocytes and one-cell embryos with pronuclei using laser scanning confocal microscopy. In metaphase II oocytes, syntaxin was located on the plasma membrane and in the cortex, where cortical granules are present, but was not seen at sites free of cortical granules. In one-cell embryos, no cytoplasmic region was free of syntaxin immunoreactivity. Immunoelectron microscopy detected syntaxin on both the plasma membrane and the vesicle membranes in mouse metaphase II oocytes. In conclusion the results indicate that syntaxin 4 co-localizes with cortical granules and participates in membrane fusion and exocytosis during the cortical reaction.

Animals↗
PubMed2003

Characteristics of F-wave in different stretched positions of the affected arm in patients with cerebrovascular diseases.

To investigate the effect of continued stretching of affected arms in patients with cerebrovascular disease (CVD), F-wave was tested from the abductor pollicis brevis (APB) during relaxation (trial 1) and 3 different continued stretches: stretched position with shoulder abduction (trial 2), stretched position with shoulder abduction and elbow extension (trial 3), and stretched position with shoulder abduction, elbow extension and wrist extension (trial 4) of the affected arm for 1 min. in 20 hemiparesis patients with moderate hypertonus (modified Ashworth scale of 2 or 3) caused by CVD (mean age of 49.5 years). Persistence and amplitude ratio of F/M in trial 2 were the same as those in trial 1. Persistence and amplitude ratio of F/M in trials 3 and 4 were significantly lower than those in trial 1. It was suggested that excitability of spinal neural function of the distal part, the APB, of the affected arm was reduced during continued stretching of the proximal part, shoulder and elbow joints, or all of the shoulder, elbow and wrist joints in hemiparesis patients with moderate hypertonus caused by CVD.

Adult↗
PubMed2002

Frequent mutations of SCN1A in severe myoclonic epilepsy in infancy.

Mutations in the neuronal voltage-gated sodium channel alpha-subunit type I gene (SCN1A) were found responsible for severe myoclonic epilepsy in infancy (SMEI). The authors describe novel mutations of SCN1A in Japanese patients with SMEI. They screened 12 unrelated patients and a pair of monozygotic twins and detected 10 mutations that lead to truncation of the protein.

Diseases in Twins↗
PubMed2002

Ruptured abdominal aortic aneurysm associated with a psoas abscess.

A case of abdominal aortic aneurysm infected by Salmonella dublin is presented. Computed tomography (CT) revealed an abdominal aortic rupture associated with a psoas abscess. An axillo-femoral bypass was performed. The aneurysm and the psoas abscess were resected.

Aneurysm, Infected↗
PubMed2002

The relationship between EMG median frequency and low frequency band amplitude changes at different levels of muscle capacity.

OBJECTIVE: To test the validity of high and low frequency band amplitudes of the surface electromyography (EMG) profile as representation of muscle fatigue. DESIGN: A within subjects (n=10) repeated measures design was used to collect surface EMG signals from the biceps during an isometric contraction under two levels of fatigue status. BACKGROUND: The use of the shift in the median frequency of the surface EMG power spectrum is a well known method of assessing muscle fatigue. Fatigue also results in amplitude changes of the specific frequency bands. The use of frequency band analysis may be an alternative option for the assessment of muscle fatigue in specific experimental settings. METHODS: Surface EMG profiles of the biceps were recorded at 1024 Hz during a sustained isometric hold at 60% of the individuals fresh and fatigued maximal voluntary isometric torque. The median frequency of the power spectrum was compared with changes in the low frequency (15-45 Hz) and high frequency (>95 Hz) bands.Results. There was a close association between median frequency shift and the amplitude of the 15-45 Hz bandwidth and the high-low frequency amplitude ratio. The association was similar for performance under different muscle capacity states. CONCLUSIONS: Frequency band amplitude analysis provides similar information to median frequency shift under isometric conditions and may be suited to specific experimental protocols in workplace fatigue studies. RELEVANCE: The use of amplitude band analysis that closely approximates the standard median frequency changes allows greater possibility of assessing muscle fatigue in different experimental settings and the use of lower sampling rates.

Adult↗
PubMed2002

Attenuation of glucan-binding protein C reduces the cariogenicity of Streptococcus mutans: analysis of strains isolated from human blood.

A blood isolate of Streptococcus mutans strain TW871 shows relatively low homology with MT8148, a reference oral isolate strain, and lacks the serotype-specific polysaccharide antigen, suggesting that other cell-surface structures correlate with cariogenicity. We compared cariogenicity of TW871 with MT8148 (serotype c) and blood isolate TW964 (serotype f) in rats. Strain TW871 showed significantly lower cariogenicity than MT8148 or TW964 and expressed significantly lower sucrose-independent cellular adhesion to saliva-coated hydroxyapatite and dextran-binding activity than strain MT8148. Strains TW871 and TW964 showed a defect in the gbpA gene by Southern hybridization analysis, while sequencing analysis revealed gbpC variation in TW871. These results suggest that variation in GbpC may alter cellular adherence properties and can be correlated with the cariogenicity of S. mutans in this strain.

Amino Acid Sequence↗
PubMed2002

Differential and quantitative analyses of mRNA expression of glucosyltransferases from Streptococcus mutans MT8148.

Streptococcus mutans produces three glucosyltransferases, coded by gtfB, gtfC, and gtfD, whose cooperative action is essential for sucrose-dependent cellular adhesion. This cellular adhesion plays an important role in the formation of dental plaque and the initiation of dental caries. Since they bear genetic similarities and are large in size, differentiation of their gene expression has been difficult, and little is known about the dynamic process of gtf expression. Using a real-time reverse-transcription/polymerase chain-reaction, we determined the expression of each gtf. Under various conditions, the relative levels of transcription were gtfB > gtfD > gtfC. Sucrose enhanced gtfD expression, whereas it reduced that of gtfB and gtfC, suggesting the presence of independent promoters. Quantitative analyses demonstrated coincidence between the ratio of expression of each gtf and the ratio previously identified as optimal for sucrose-dependent adhesion in vitro, suggesting that S. mutans produces GTF at an optimal ratio to adhere to the tooth surface.

Analysis of Variance↗
PubMed2002

Characteristic appearances of the H-reflex and F-wave with increased stimulus intensity in patients with cerebrovascular disease.

We experienced H-reflex may be evoked with supramaximal stimulation in patients with cerebrovascular disease (CVD). In this study, we investigated the relationship between the characteristic appearances of H-reflex and F-wave with increased stimulus intensity and neurological signs. We examined the H-reflex and F-wave of the affected arm with increased stimulus intensity during muscle relaxation in 31 patients (17 males and 14 females) with hemiparesis caused by CVD. Mean patient age was 56.0 (range 30-82) years. 30 healthy subjects, mean age of 56.2 (range 28-80) were investigated using the same method as controlled group. H-reflex and F-wave with increased stimulus intensity after the median nerve stimulation at the wrist were recorded from the opponence pollicis muscle on the affected side in patients with CVD or right arm in the healthy subjects. Appearance patterns of the H-reflex and F-wave with increased stimulus intensity was separated into four types. Type 1: F-wave appeared with increased stimulus intensity, but there was no H-reflex. Type 2: H-reflex and F-wave both appeared with increased stimulus intensity, but the F-wave followed disappearance of the H-reflex with increased stimulus intensity. Type 3: H-reflex and F-wave both appeared with increased stimulus intensity, but the F-wave appeared during the H-reflex with increased stimulus intensity. Type 4: Only the H-reflex appeared with increased stimulus intensity, but there was no F-wave. Neurological findings including muscle tone and tendon reflex were also evaluated. Findings on muscle tone and tendon reflex were classified into increased (markedly, moderately and slightly), normal and decreased. Results were analyzed as follows; 1) The characteristic appearances of H-reflex and F-wave in the healthy subjects and 2) The relationship between characteristic appearances of waves with increased stimulus intensity and neurological signs in patients with CVD. 1) Pattern of the H-reflex and F-wave with increased stimulus intensity in all healthy subjects were type 1. 2) Patterns of the H-reflex and F-wave with increased stimulus intensity in patients with markedly increased muscle tone and tendon reflex were almost always type 4. Those with moderately increased signs in this parameter demonstrated type 2 or 3. Those with slightly increased signs in this parameter demonstrated type 1 or 2. Those with normal or decreased signs were type 1. These findings suggest that the characteristic appearances of the H-reflex and F-wave was influenced by grade of neurological signs.

Adult↗
PubMed2002

Sequential variation of groundwater quality in an agricultural area with greenhouses near the coast.

The quality of coastal groundwater utilized for greenhouse-farming is liable to be affected by not only fertilization practices but also seawater intrusion into the aquifer. In this study, the sequential variations of groundwater quality and the mechanisms of its pollution processes were analyzed through successive field measurements from June 2000 to May 2001 in such a coastal agricultural area located in Kochi prefecture, Japan. The analysis revealed that the ions for NO3-, SO4(2-), Mg2+ and Ca2+ in the groundwater originated primarily from the ground surface due to the seasonal flooding activities. It was also estimated that denitrification occurred at three well points, and the rate constants were 0.010, 0.019 and 0.089 d-1, respectively. Na+, Mg2+ and Ca2+ concentrations in the groundwater governed by seawater intrusion phenomena showed strong correlations with the concentration of Cl- by excluding the influence of fertilizer. Cation exchange phenomena between Ca2+ and Na+ and as well as the mixing with seawater had a considerable effect on the groundwater quality.

Agriculture↗
PubMed2002

[Reoperative off-pump coronary artery bypass surgery for restenosis after percutaneous coronary intervention as a part of hybrid revascularization chronic renal failure on hemodialysis; report of a case].

60-year-old man on chronic hemodialysis had hybrid revascularization. Percutaneous coronary intervention (PCI) for right coronary artery (RCA) combined with left internal mammary artery-left anterior descending artery anastomosis through left anterior small thoracotomy were performed. But, frequent re-stenosis after PCI necessitated 4 repeated PCI. Finally, the patient agreed to re-do, and underwent reoperative off-pump coronary artery bypass through midsternotomy. Right internal mammary artery-saphenous vein lengthened composite graft was successfully anastomosed to heavily calcified RCA without heart-lung machine. Strategy of revascularization for hemodialysis patient was discussed.

Angina, Unstable↗