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Biomedical subjects

H Yoshida

Publications and source records attributed to H Yoshida.

At least 883 records · Page 49Linked to original sources

Endoscopic variceal ligation using a clipping apparatus in children with portal hypertension.

BACKGROUND AND STUDY AIMS: Endoscopic variceal sclerotherapy (EVS) is a procedure frequently used to control bleeding from esophageal varices in adults. The use of EVS has been restricted in children due to the associated risks. The present paper describes the safety and efficacy of a new apparatus for endoscopic variceal ligation with clipping (EVLC). PATIENTS AND METHODS: Endoscopic variceal ligation using a clipping apparatus (HX-3 L, Olympus, Tokyo, Japan), was carried out in 12 children with portal hypertension (four with extrahepatic portal vein obstruction and eight with congenital biliary atresia; four boys and eight girls, age range 2-14 years). A total of 417 variceal clipping procedures were carried out in 56 separate EVLC sessions; 118 esophageal varices and 65 gastric fundic varices were ligated using 417 clips. Initially, the EVLC sessions were repeated at approximately monthly intervals until the varices were eradicated, but the interval was gradually extended to twice a year or once a year. RESULTS: Varices were successfully ligated at the first attempt in 414 (99.3%) of the 417 clipping procedures. The esophageal varices were initially eradicated in all patients. The follow-up period ranged from nine to 48 months, and complete variceal eradication or reduction of the grade of the varices was obtained in 11 of the 12 patients after initial eradication during this period. CONCLUSIONS: These data indicate that EVLC offers advantages in the treatment of patients with esophageal varices combined with gastric fundic varices.

Adolescent↗

Effect of electric pulses and antiproliferative drugs on cultured bovine retinal pigment epithelial cells.

PURPOSE: The effect of antiproliferative agents with high voltage electric pulses (EP) on the proliferation of retinal pigment epithelial (RPE) cells was investigated. METHODS: Bovine RPE cells were cultured and exposed briefly to an antiproliferative agent: 0.005 to 500 microM of bleomycin (BLM), 0.005 to 500 microM of mitomycin C (MMC) or 0.05 to 5000 microM of 5-fluorouracil (5-FU) with or without high voltage electric pulses (EP: 2,000V/cm, 98 microseconds, 8 pulses). Streptomycin (SM) was used as a negative control. Cell proliferation with or without antiproliferative agents was assessed on day 3. DNA fragmentation was assessed by flow cytometric analysis. RESULTS: Treatment with BLM, MMC or 5-FU inhibited cell proliferation in a dose-dependent manner, with and without EP (p < 0.05). EP enhanced the inhibitory effect of BLM as much as 3,000 times (ID50: BLM without EP; 163.7 microM, BLM with EP; 0.0574 microM, MMC without EP; 132.4 microM, MMC with EP; 26.2 microM, 5-FU without EP; 616.4 microM, 5-FU with EP; 873.8 microM). EP treatment showed an inhibitory effect of BLM on cell proliferation more prominently than BLM alone. The cell numbers of 0.5 microM BLM treatment without EP were 87.4 +/- 11.7% (mean +/- SD), whereas the cell numbers of 0.5 microM BLM with EP were 21.1 +/- 2.16% (p < 0.005). BLM treatment with EP increased the apoptosis like DNA fragmentation in the flow cytometric DNA histogram, showing dominant accumulation in the A0 region, which is the population of DNA fragmentation. (The cell population of the A0 region: control, 3.4%; EP alone, 18.9%; BLM treatment without EP, 32.0%; with EP, 93.8%.) CONCLUSIONS: EP treatment enhances the inhibitory effect of BLM on RPE cell proliferation. The combination of electric pulse and BLM treatment can localize the drug effect and reduce the necessary dose of the antiproliferative agent in comparison to BLM treatment alone.

Animals↗

Olive oil increases the magnitude of postprandial chylomicron remnants compared to milk fat and safflower oil.

OBJECTIVE: The acute effects of olive oil, milk fat and safflower oil on postprandial lipemia and remnant lipoprotein metabolism were investigated. METHODS: Eight Healthy male volunteers randomly underwent three types of oral fat-vitamin A loading tests. The test drink was a mixture of retinyl palmitate (RP)(50,000 IU of aqueous vitamin A/m2 body surface area) and one of the three types of oils (40 g of fat/m2 body surface area): olive oil (70.7% oleic acid of total fatty acids); milk fat (69.3% saturated fatty acid); safflower oil (74.2% linoleic acid). RESULTS: Olive oil significantly increased plasma triacylglycerol and RP concentrations 4 hours after fat loading, as compared to other fats. Increases of remnant like particle concentrations were higher after olive oil than after the other two fats. CONCLUSION: These results show that olive oil increases the magnitude of postprandial chylomicrons and chylomicron remnants compared to milk fat and safflower oil.

Adult↗

NMR characterization of physicochemical properties of rat D-dopachrome tautomerase.

D-dopachrome tautomerase (DOPD) catalyzes the conversion of D-dopachrome to 5,6-dihydroxyindole. DOPD was found to have amino acid sequence homology with macrophage migration inhibitor factor (MIF), suggesting that DOPD functions as a proinflammatory cytokine. We here demonstrate the physicochemical properties of DOPD by nuclear magnetic resonance (NMR). The native molecular weight of rat DOPD was about 37 kDa as calculated from Sephadex G-100 column chromatography. Since the deduced molecular weight of its cDNA (117 amino acid residues) is 12.5 kDa, it is assumed that DOPD exists as a homotrimer in native form. Since several methyl proton resonances were observed in the high magnetic field region of the one-dimensional 1H-NMR spectrum, DOPD appeared to have a hydrophobic core in which methyl groups and aromatic groups are located close to each other. From a simple integration of one-dimensional 1H-NMR spectra, the contents of the alpha-helix, beta-strand, and random coil were calculated to be 16%, 68%, and 16%, respectively. Since the denaturation temperature of DOPD is exceedingly high at the range between 90-100 degrees C, it is considered that the high beta-strand content may contribute to its heat stability.

Animals↗

Two major strains of type 1 wild poliovirus circulating in Indochina.

Two hundred ninety-four isolates from 329 patients with acute flaccid paralysis in Cambodia and Vietnam during 1992-1995 were identified as type 1 wild polioviruses. Among these isolates, 85 were selected as geographic representatives and were examined by determining the nucleotide sequences of their genome in the VP1 region. The phylogenic analysis revealed that all of the isolates examined were classified into groups A and B. Isolates belonging to group A had been found only in northern Vietnam until 1993 but not in 1994 and 1995. Group B isolates were located in both northern and southern Vietnam and Cambodia. In 1994 and 1995, however, only group B isolates were found in the Mekong Delta area in southern Vietnam and Cambodia. Isolates of groups A and B were genetically different from strains previously isolated in other Asian countries. One of the two indigenous wild polioviruses still remains to be eliminated in this area.

Antigens, Viral↗

Effect of tea flavonoid supplementation on the susceptibility of low-density lipoprotein to oxidative modification.

Dietary flavonoid intake has been reported to be inversely associated with the incidence of coronary artery disease. To clarify the possible role of tea flavonoids in the prevention of atherosclerosis, we investigated the effects of tea flavonoids on the susceptibility of low-density lipoprotein (LDL) to oxidative modification. In an in vitro study, catechins or theaflavins (25-400 mumol/L) were added to plasma and incubated for 3 h at 37 degrees C. Then, the LDL fraction was separated by ultracentrifugation. The oxidizability of LDL was estimated by measuring conjugated diene, thiobarbituric acid-reactive substances (TBARS), and lipid peroxides after cupric sulfate was added. TBARS and lipid peroxides in the supernates were also measured after incubation with macrophages. Catechins significantly (P < 0.01 by ANOVA) and dose-dependently prolonged the lag time before initiation of oxidation. Among the catechins, epigallocatechin gallate exerted the most marked effect, prolonging the oxidation lag time more than vitamin E at the same molar concentration. Theaflavins exerted stronger inhibitory effects than catechins. Macrophage-mediated LDL oxidation was also inhibited by adding these tea flavonoids to the plasma samples. In an in vivo study, 14 healthy volunteers consumed 750 mL black tea/d for 4 wk. After the subjects had consumed tea for 4 wk, the lag time before LDL oxidation was significantly (P < 0.01) prolonged from 54 to 62 min. This minor prolongation occurred despite much lower plasma flavonoids than were used in vitro. No significant change was observed in eight control volunteers. LDL exposed to tea flavonoids in vitro or in vivo reduced oxidizability. We speculate that tea flavonoids may have a role in ameliorating atherosclerosis.

Adult↗

Three distinctive steps in Peyer's patch formation of murine embryo.

Investigation of the process of Peyer's patch (PP) formation has been hampered by difficulties in identifying its initial step in the embryo. In this study, we overcame this problem by means of whole-mount immunohistochemistry using mAb against the molecules which were expressed in the cells accumulated at the site of PP development. This method was sensitive enough to distinguish a minute cell cluster in the developing gastrointestinal tract. We analyzed the time course of the expression of various surface markers and found that PP formation proceeds through three successive steps. The first is the appearance of a VCAM-1+ cell cluster at 15.5 days postcoitus (d.p.c.). Histological examination of the VCAM-1+ clusters suggested that VCAM-1+ cells represent a stromal component. The second step is characterized by the accumulation of round cells expressing la, IL-7R or CD4 at 17.5 d.p.c. Lymphocytes expressing CD3 or B220 were detected only in the final step which started at 18.5 d.p.c. Using any of these markers, the aggregation was initially detected on the upper jejunum and it extended to the colon as the number of clusters increased. At the neonatal stage, the number reached up to eight or nine, irrespective of the antibodies used for the detection. In the aly/aly mutant mouse, where no lymph nodes or PP are found in the adult, none of these three steps was detected. On the other hand, in the SCID mouse that is defective in the formation of mature lymphocytes, the first and second step proceeded, whereas the third step was undetectable. These findings suggest that the progression of each step is indeed regulated by different mechanisms.

Animals↗

Studies on the SEM examination of un-coated non-conductive specimens prepared by plasma-ion shower method.

The plasma-ion shower method is an effective technique for the SEM examination of un-coated electrically non-conductive specimens. However, milling damage to specimens cannot be prevented by the irradiation method using accelerated Ar ion beam. We, therefore, applied positive ions obtained by plasma discharge to the specimen like a shower by reflecting from the target electrode surface. He, N2 or Ar was used as the environmental gas, and the discharge voltage was varied from 200 V to 1000 V. At 5 kV to 25 kV of SEM accelerating voltage, it was possible to visualize un-coated hard and soft biological and inorganic specimens under 5 mA or 10 mA of discharge current and 5 min or 10 min of processing time.

Adult↗

Hepatocyte growth factor stimulated proliferation, migration, and lumen formation of human endometrial epithelial cells in vitro.

Human endometrial regeneration has been considered to be modulated by several growth factors. However, little is known about the detailed mechanisms involved in the repair of the endometrium during the menstrual period. Hepatocyte growth factor (HGF) is a pleiotropic growth factor that reportedly acts on various epithelial cells. In the present study, we observed HGF and mRNA expression in human endometrium and mRNA expression in cultured endometrial epithelial cells of the c-met gene by reverse transcription-polymerase chain reaction and Southern blot hybridization. We also examined the biological role of HGF on the regeneration of the endometrium by using cultured endometrial epithelial cells in their proliferative phase. With the proliferation assay, the addition of 10-50 ng/ml of HGF showed that HGF was mitogenic in a dose-dependent manner. With Boyden's chamber technique, 50 ng/ml of HGF significantly stimulated cell migration. In a three-dimensional cell-culture system, the endometrial epithelial cells formed cell clusters and gradually formed epithelial lumens, both of which were stimulated by 50 ng/ml of HGF. Results suggest the HGF stimulates the proliferation and migration of, and morphogenic changes in, endometrial epithelial cells. HGF may modulate the regeneration of the endometrium during menstruation.

Adult↗

Chromosomal translocations in human soft tissue sarcomas by interphase fluorescence in situ hybridization.

In soft tissue sarcomas, clonal rearrangement of chromosomes has been shown by cytogenetic analysis to be unique and specific for tumor types. The development of fluorescence in situ hybridization (FISH) has allowed detection of chromosomal rearrangements in the interphase nuclei isolated from paraffin-embedded tissues. Three kinds of translocations in the interphase nuclei that were isolated from 47 cases of soft tissue sarcomas were examined by FISH with chromosome-specific DNA probes of centromeric and total probes. Of 47 soft tissue sarcomas 42 (89.4%) revealed tumor-specific translocations by retrospective cytogenetic analysis. Translocation t(X;18) was detected in 25/28 synovial sarcomas; translocation t(11;22) in 5/6 Ewing's sarcomas and primitive neuroectodermal tumors (PNET); and translocation t(12;16) was found in 12/13 liposarcomas, including 10 myxoid and two round cell types as clonal chromosomal aberrations specific for both subtypes. Based on the cytogenetic analysis, Ewing's sarcoma is related closely with PNET as shown by MIC2-protein reactivity. Other cytogenetic findings of translocation t(12;16) indicate that round cell liposarcomas share chromosomal changes with myxoid liposarcomas, and further suggest that both tumor subtypes of liposarcoma may possess common precursor cells. FISH is a useful aid in determining the tumor type of soft tissue sarcomas with regard to histogenetic origin.

Adolescent↗

Immunohistochemical distribution of lymph capillaries and blood capillaries in the synovial membrane in cases of internal derangement of the temporomandibular joint.

The distribution of lymph capillaries and blood capillaries in the synovial membrane was examined immunohistologically with anti-human collagen IV antibody and anti-human von Willebrand factor in 26 human temporomandibular joint (TMJ) samples comprising discs with adjoining synovial membrane from 10 control TMJs and from 16 TMJs with internal derangement. Three different distribution types were observed in the synovial membrane. In the control samples, the occurrence of blood capillaries and lymph capillaries was rare. In mildly hyperplastic synovitis, lymph capillaries were observed just beneath the surface of the synovial membrane, whereas blood capillaries occurred in a little deeper layer of the synovial membrane. In a severely hyperplastic synovitis, both lymph and blood capillaries were observed frequently. The present results suggest that the different distribution patterns of lymph capillaries and blood capillaries reflect the degree of synovitis but can not be attributed to specific clinical symptoms.

Adult↗

UVB DNA dosimeters analyzed by polymerase chain reactions.

Purified bacteriophage lambda DNA was dried on a UV-transparent polymer film and served as a UVB dosimeter for personal and ecological applications. Bacteriophage lambda DNA was chosen because it is commercially available and inexpensive, and its entire sequence is known. Each dosimeter contained two sets of DNA sandwiched between UV-transparent polymer films, one exposed to solar radiation (experimental) and another protected from UV radiation by black paper (control). The DNA dosimeter was then analyzed by a polymerase chain reaction (PCR) that amplifies a 500 base pair specific region of lambda DNA. Photoinduced damage in DNA blocks polymerase from synthesizing a new strand; therefore, the amount of amplified product in UV-exposed DNA was reduced from that found in control DNA. The average lesion frequency per 500 base pair per strand at 16 PCR cycles was approximately 1.22, 1.00, 0.70 and 0.50 for 30 ng, 50 ng, 100 ng and 150 ng of dried DNA, respectively, after a total dose of 60 kJ m(-2) delivered with a solar UVB simulator. Although the average lesion frequency increases linearly with increasing doses for four different amounts of template DNA, the lesion frequency seems to be averaged by the amplified products from the protected lambda DNA molecules below the top few layers. The average daily dose, equivalent to the UV dose applied with the solar UVB simulator was 10.2 +/- 0.4 kJ m(-2) with the 50 ng containing DNA dosimeter in September 1995 in Melbourne, FL. Both 50 ng and 150 ng containing DNA dosimeters produced the same average daily dose within experimental error in January 1996, which was 5.2 +/- 0.3 kJ m(-2) at the same location. The dried lambda DNA dosimeter is compact, robust, safe and transportable, stable over long storage times and provides the total UVB dose integrated over the exposure time.

Australia↗

Solar UVB dosimetry by amplification of short and long segments in phage lambda DNA.

DNA UVB dosimeters, consisting of minidots of dried bacteriophage DNA placed on a UV-transparent polymer film, were analyzed by polymerase chain reactions (PCR). Ultraviolet-B dosimetry obtained with amplification of phage lambda DNA of segments of 1.08 kilobase pairs (kbp) and 2.24 kbp was compared with that obtained with amplification of a 0.5 kbp segment (H. Yoshida and J. D. Regan, 66, 82-88, Photochem. Photobiol. 1997). The number of lesions in each segment induced by UV radiation is proportional to the size of the amplified segments; thus, the average lesion frequency per unit dose was greatest in the 2.24 kbp and least in the 0.5 kbp segment. The average lesion frequency per unit dose was approximately 3.5 x 10(-2) and approximately 11.9 x 10(-2) m2 kJ-1 for 1.08 kbp and 2.24 kbp, respectively, compared to that for 0.5 kbp of approximately 1.7 x 10(-2) m2 kJ-1 at 50 ng DNA. Dependability of DNA dosimeters, containing 50 ng and 100 ng, was tested by placing the DNA dosimeters for a time period of either 1 or 2 days outdoors on 8-12 January at Melbourne, FL. The daily dose was obtained directly with amplification of the 2.24 kbp segment and the 2 day continuous dose was obtained with amplification of the 1.08 kbp segment. Although the average lesion frequencies were different, both 50 ng and 100 ng DNA dosimeters provided about the same UVB dose, equivalent to the dose applied with a solar UVB simulator. The total UVB dose for 4 days obtained by amplification of the 1.08 and 2.24 kbp segments was 19.4-20.8 kJ m-2, which is within experimental error with the 4 day continuous dose obtained with 0.5 kbp segments. The average daily dose obtained by 0.5 kbp and 1.08 kbp agreed with the average daily dose directly obtained with 2.24 kbp.

Bacteriophage lambda↗

Osmotic flow transients during acetylcholine stimulation in the perfused rat submandibular gland.

Osmotic stress was applied to the perfused rat submandibular gland during steady-state fluid secretion. Alterations of perfusate osmolarity, by addition or withdrawal of sucrose or NaCl, caused transient changes in secretory rate during continuous stimulation with 1 microM acetylcholine (ACh). Hyposmotic perfusates transiently increased, and hyperosmotic perfusates transiently reduced, the secretory rate. The transients were attributed to changes in osmotic flow resulting from changes in the instantaneous transepithelial osmotic gradient. The time course of the change in interstitial osmolarity was determined by using a Cl- electrode to record the changes in interstitial Cl- concentration following a step change in perfusate Cl- concentration. From the calculated changes in interstitial osmolarity and the resulting changes in secretory rate, the osmotic water permeability of the secretory pathway was estimated to be greater than 15.0 +/- 1.2 microliter (mosmol 1-1)-1 min-1 (g wet weight)-1 (9.8 x 10(-6) +/- 0.8 x 10(-6) l atm-1s-1g-1). The transepithelial gradient required to sustain steady state, ACh-evoked secretion would therefore be less than 16 mosmol l-1 NaCl, which is consistent with previous micropuncture data indicating that the luminal fluid is approximately isosmotic.

Acetylcholine↗

[A case of cat scratch disease identified by an elevated Bartonella henselae antibody level using enzyme immunoassay].

A 68-year-old male was admitted to our hospital because of fever and a 2-week history of inguinal adenomegaly. Since he owned a cat, cat scratch disease was suspected. But it was necessary to distinguish cat scratch disease from lymphoma type adult T-cell leukemia because he showed a high level of antibody against HTLV-1. An excisional biopsy of the inguinal node was performed. Histopathologic examination revealed abscess-forming granulomatous lymphadenitis compatible with cat scratch disease. A Warthin-Starry silver stain showed pleomorphic bacilli in the lymph node. So we confirmed a serological response to Bartonella henselae, the causative agent of cat scratch disease, using enzyme immunoassay (EIA). The IgG antibody level to B. henselae was positive at 42 EIA Unit before treatment. After treatment with intravenous cefepime and oral tosufloxacin, his physical symptoms improved and the antibody level decreased to less than 12 EIA Unit. EIA was very useful for diagnosis of this case. Serology to B. henselae may replace traditional diagnostic criteria for cat scratch disease.

Aged↗

Cloning and sequencing of a novel gene (recG) that affects the quinolone susceptibility of Staphylococcus aureus.

In a study of the quinolone resistance genes in Staphylococcus aureus, a recG homolog was cloned as a gene affecting quinolone susceptibility. Sequencing analysis revealed that the gene consists of 2,061 nucleotides and encodes a 686-amino-acid polypeptide, which shows 38, 39, and 50% amino acid identity with the RecGs of Escherichia coli, Haemophilus influenzae, and Streptococcus pneumoniae, respectively. Seven helicase motifs are well conserved in the gene product. A plasmid carrying the gene complemented a recG-deficient mutant of E. coli with respect to mitomycin hypersusceptibility, demonstrating that the gene product is functionally equivalent to E. coli RecG. These results indicate that the gene is the recG gene of S. aureus. S. aureus RCM101 (recG::Tn551), designated S. aureus 3f33, is four to eight times more susceptible to quinolones than the parent strain, RCM101. The transformation of strain 3f33 with a plasmid carrying the S. aureus recG gene made it as quinolone resistant as strain RCM101. These results suggest that the recG gene is involved in the repair of DNA damage resulting from quinolone treatment in S. aureus.

4-Quinolones↗

High prevalence of cytotoxin positive Helicobacter pylori in patients unrelated to the presence of peptic ulcers in Japan.

BACKGROUND: It has been reported that infection with vacuolating cytotoxin positive Helicobacter pylori strains is associated with gastroduodenal disease in Western countries. AIMS: To evaluate the prevalence of cytotoxin producing strains among patients with H pylori infection in relation to gastrointestinal diseases in Japan. PATIENTS: Ninety seven patients undergoing endoscopy. METHODS: A Western blot assay was conducted to detect serum antibodies against the cytotoxin using recombinant cytotoxin (VacA protein) as an antigen. To obtain a purified recombinant cytotoxin, the vacA gene (2233 nucleotides) was cloned into an expression vector to produce the protein (744 amino acids), which was expressed in Escherichia coli. RESULTS: Serum IgG antibodies to the cytotoxin were present in 85%, 95%, 95%, and 100% of infected patients with gastric ulcer (n = 26), duodenal ulcer (n = 21), chronic gastritis (n = 19), and endoscopically normal mucosa (n = 14), respectively. CONCLUSION: The western blot method using recombinant VacA protein is simple and useful for detecting antibody to vacuolating cytotoxin. This method showed antibodies against cytotoxin were highly prevalent, even in subjects with endoscopically normal mucosa in Japan, indicating that the cytotoxin may not be an independent cause of gastrointestinal diseases induced by H pylori infection.

Amino Acid Sequence↗