Biomedical subjects

Y Sakai

Publications and source records attributed to Y Sakai.

At least 55 records · Page 3Linked to original sources

PubMed2002

Growth-dependent and species-specific accumulation of polychlorinated biphenyls (PCBs) in tidal flat organisms collected from the Ariake Sea, Japan.

The growth-related as well as species-specific accumulation of polychlorinated biphenyls (PCBs) was studied for tidal flat organisms collected from the Ariake Sea, western Japan. Elevated concentrations of PCBs were found in omnivore fishes, followed in decreasing order by crabs, herbivore fishes, and mussels. This revealed that trophic levels play an important role in PCB accumulation in these organisms. Age- and body length-dependent accumulations of PCBs were observed in herbivorous mudskippers, although a large range of concentrations was found in similar growth stage of fishes. High correlations have been found between concentrations and body length rather than age, which may indicate that the growth rate, which is strongly influenced by the feeding rate of diets, seems to be the predominant factor in determining PCB accumulation. Besides, based on PCB levels in eggs and the whole body in herbivore fishes, the transfer rate of PCBs was estimated to be approximately 10% of female body burdens. Comparison of PCB compositions between eggs and whole body suggests the selective transfer of lower-chlorinated congeners to eggs, which may be due to their instantaneous periods to achieve steady state between egg and whole body lipids. The relationship between BSAFs (biota-sediment accumulation factors) in organisms and log Kow revealed that omnivore mudskipper significantly accumulated PCBs in their body, which might be due to their greater feeding rate and/or higher trophic status in the tidal flat environment.

Animals↗

PubMed2002

CD34+ stromal cells and hyalinized vascular changes in the anal fibroepithelial polyps.

AIMS: To elucidate the pathogenesis of the anal fibroepithelial polyp, we examined surgically resected lesions histopathologically. METHODS AND RESULTS: Twenty-seven surgically resected anal fibroepithelial polyps were investigated histologically with an additional immunohistochemical examination using anti-CD34. For a control study, the surgical specimens of the anal canal showing non-polypoid lesions, obtained from haemorrhoidectomy (18 specimens) and rectectomy (five specimens) due to rectal cancer without anal canal involvement, were also analysed. We demonstrated characteristic spindle or stellate cells immunohistochemically positive for CD34 in the anal fibroepithelial polyps (24/27, 89%). The number of CD34+ cells was statistically related to the size of anal fibroepithelial polyps, although CD34+ stromal cells were recognized in the non-polypoid anal submucosa and haemorrhoids. We also found hyalinized vascular changes in the base of six anal fibroepithelial polyps examined. These features were not detected in the non-polypoid anal canal. CONCLUSIONS: An increase in CD34+ stromal cells may play a role in the enlargement of anal fibroepithelial polyps. CD34+ stromal cells are suggested to be distinctive mesenchymal cells with a capability for tissue repair and overgrowth. The vascular impairment could be secondary change associated with localized tissue damage by abnormal traction.

Adult↗

PubMed2002

Changes in sodium transport in epithelial cells in a rat ileal-augmented bladder model.

OBJECTIVE: To investigate changes in the morphology and sodium transport ability of intestinal epithelium diverted to the urinary tract, using an in vitro sodium-binding benzofuran isophthalate (SBFI) technique, as the effects of long-term urine exposure on the transport of electrolytes through intestine are incompletely understood. MATERIALS AND METHODS: Ileal augmentation cystoplasty was conducted in female Sprague-Dawley rats; at 3 and 12 months after surgery the serum concentration of sodium, chloride and potassium were measured. Sodium transport in the ileal epithelial cells diverted to the urinary tract was evaluated using SBFI, as the value of the 340/380 nm excitation ratio measured with fluorescence spectrophotometry. The villous height and the number of villi per ileal length were obtained from haematoxylin and eosin-stained sections. RESULTS: After 3 months the mean (sd) serum sodium concentrations in normal and augmented rats were 140.4 (2.5) and 140.7 (3.5) mmol/L, respectively; the chloride concentration in normal rats was 97.0 (2.9), and in augmented rats at 3 and 12 months it was 102.4 (2.9) and 99.0 (3.7) mmol/L, respectively. At 3 months, chloride concentrations were significantly higher in augmented than in normal rats (P < 0.05). The mean (sd) 340/380 nm ratio increased by 0.85 (0.09) in the normal ileum, and by 0.73 (0.15) and 0.49 (0.23) in the ileum of augmented rats at 3 and 12 months, respectively; the difference between normal and augmented ileum at 12 months was significant (P < 0.05). At 12 months the villous height in the augmented ileum, at 227.6 (16.0) micro m, was significantly less than in the normal ileum, at 803.4 (66.2) micro m (P < 0.05). However, the number of villi/mm ileum in normal and augmented rats at 12 months was 13.7 (1.5) and 15.0 (0.8), respectively, and not significantly different. CONCLUSION: Sodium transport decreased significantly after long-term exposure to urine; the improvement in metabolic change was probably attributable to alterations of electrolyte transport and atrophic changes of the villus.

Animals↗

PubMed2002

Successful repair of a damaged duodenal stent by cutting stent wires and placement of a second stent.

Duodenal stenting has been gradually established in recent years because it is less invasive than standard surgical procedures and produces a rapid therapeutic response. For palliation of both duodenal and biliary stenoses, double stenting may be performed. Duodenal stents offer a great advantage in allowing endoscopic retrograde cholangiopancreatography (ERCP) without the need for balloon dilation. When biliary stent dysfunction occurs, the patient undergoes diagnostic and/or therapeutic ERCP across the duodenal stent. We encountered a duodenal stent fracture in a patient who required repeated ERCPs for stent dysfunction. Duodenal stent fractures have not previously been reported. The damaged stent was successfully repaired by using a cutting wire filament and placing another duodenal stent coaxially with the first. Clinicians should be aware of the possibility of stent fracture following endoscopic procedures, such as an ERCP, that require passage through the stent. The procedure described in this report would be of significant benefit if a gastrointestinal stent is fractured and occluded by a broken part.

Aged↗

PubMed2002

Technical modifications and strategies for stenting gastric outlet strictures using esophageal endoprostheses.

BACKGROUND AND STUDY AIMS: The outcome of stenting gastric outlet stricture is favorable compared with a bypass operation which has significant morbidity and mortality. In Japan, this procedure is particularly complicated by a lack of enteral stents. We report some technical stratagems for stent placement for gastric outlet strictures. PATIENTS AND METHODS: Between February 1993 and July 2001, 23 patients with gastric outlet strictures (14 men, nine women; mean age 72 years) underwent stent placement using an esophageal stent system. The Ultraflex or Z-stents were used in 18 or five patients, respectively. With the Ultraflex, we increased the length of the delivery system. Some patients underwent stent placement with the help of endoscopic assistance with a grasping forceps or a home-made sheath. RESULTS: The metal stent was successfully inserted in all patients. There were no complications during the procedure. Migration occurred in two out of five patients treated with the Z-stent, whereas there was no migration in patients treated with the Ultraflex stent. In two patients, curable pancreatitis was caused by pressure on the duodenal papilla. One of these patients also experienced bile stasis which required biliary decompression. There were three cases of obstruction, caused by tumor ingrowth (1), hyperplasia (1) and stent fracture (1); recanalization by an additional stent placement and/or cutting stent filaments was successful. All the patients died, with a median survival period of 52 days. There was no procedure-related mortality. CONCLUSIONS: With some technical modification, stent placement for gastric outlet stricture, even using an esophageal stent, is feasible. This procedure offers good palliation with no major complications.

Adult↗

PubMed2002

A new cannula with a flexible tip (Swing Tip) may improve the success rate of endoscopic retrograde cholangiopancreatography.

BACKGROUND AND STUDY AIMS: Endoscopic retrograde cholangiopancreatography (ERCP) is sometimes unsuccessful because the anatomy of the papilla of Vater precludes cannulation by routine means. The efficacy of a new flexible tip cannula (the Swing Tip) was studied in patients in whom routine ERCP was unsuccessful. PATIENTS AND METHODS: The Swing Tip cholangiographic catheter has an articulation at its tip which allows the tip to be flexed from 90 degrees to 30 degrees. Routine cholangiography was successful in 175 of 195 patients in whom it was attempted between September 2000 and November 2001. ERCP with the Swing Tip catheter was attempted in the 20 patients in whom ERCP had failed. RESULTS: In 17 of 20 patients, we attempted to insert the Swing Tip catheter into the common bile duct to perform cholangiography. Insertion was successful in 11 patients and unsuccessful in six. There were no complications related to the procedure. CONCLUSION: The Swing Tip catheter is a useful adjunct to standard ERCP catheters for patient in whom standard techniques are unsuccessful.

Bile Duct Diseases↗

PubMed2002

Helicobacter pylori infection in rheumatoid arthritis: effect of drugs on prevalence and correlation with gastroduodenal lesions.

OBJECTIVE: The aim of this study was to investigate the impact of Helicobacter pylori infection on clinical features in patients with rheumatoid arthritis (RA) under medication with non-steroidal anti-inflammatory drugs. METHODS: One hundred and eighty-four patients with RA were tested for the presence of H. pylori infection. Clinical features and gastroduodenal lesions were compared between H. pylori-positive and -negative patients. RESULTS: One hundred and thirteen patients were positive and 71 patients were negative for H. pylori. The age, severity of RA, prevalence of gastrointestinal symptoms and gastroduodenal lesions and the class of gastroprotective drugs were not different between the two groups. Reflux oesophagitis was less frequent and sulphasalazine was less frequently administered in the H. pylori-positive group. CONCLUSIONS: The severity of RA, prevalence of gastroduodenal lesions other than reflux oesophagitis and the application of gastroprotective drugs do not seem to depend upon H. pylori infection in RA patients. Sulphasalazine may be protective against H. pylori infection.

Adult↗

PubMed2002

Catalytic ozonation of an organophosphorus pesticide using microporous silicate and its effect on total toxicity reduction.

Catalytic ozonation is promising as one of the advanced oxidation processes because of its effective use of ozone and its improved treatability of organic compounds through radical reactions. In this article, we investigated the feasibility of microporous silicates as a potential catalyst for a catalytic ozonation process. Organophosphorus insecticide, dichlorvos (DDVP), was employed as a model chemical for assessing conventional ozonation and catalytic ozonation, because its oxidative intermediate is toxic and is not degraded by direct ozonation. It was found that ozone was well adsorbed and simultaneously decomposed in microporous silicates, resulting in the production of possible radical species. In the presence of microporous silicates, radical reactions by decomposed ozone were evidenced by phosphate ion release that shows a degradation of a toxic intermediate of DDVP. Accordingly, cytotoxicity was successfully decreased. In a continuous treatment process combining a conventional ozonation vessel and a microporous silicate column for an effective use of residual ozone, enhanced degradation of DDVP was demonstrated by a decrease of DOC, an increase of the PO4(3-) concentration and reduction in the cytotoxicity. This new treatment mechanism is likely to be promising as an advanced water treatment process particularly when we think about better toxicity reduction of wastewaters.

Adsorption↗

PubMed2002

Enhanced in vitro maturation of fetal mouse liver cells with oncostatin M, nicotinamide, and dimethyl sulfoxide.

Although cells isolated from fetal liver are one of the major sources for liver tissue engineering, it is still very difficult to induce them to fully differentiate in vitro into mature hepatocytes. We therefore investigated the effects of nicotinamide (NA), dimethyl sulfoxide (DMSO), and oncostatin M (OSM) on differentiation in terms of the expression of various liver-specific functions, because these factors have been reported to induce the emergence of possible hepatocyte progenitor cells (small hepatocytes) in adult rat hepatocyte culture or maturation of fetal mouse liver cells in culture. Fetal liver cells isolated from mouse embryos were cultured for 5 weeks in collagen-precoated plates. NA (10 mM) and DMSO (1%) remarkably enhanced the emergence of small hepatocytes, and OSM also synergistically enhanced the selective growth of small hepatocytes and inhibited the growth of blood cell populations. In the presence of these three factors, such small hepatocytes became dominant in culture, so that they covered almost 60-70% of confluence after week 2. In addition, some of them piled up over the small hepatocyte monolayer and displayed distinctively differentiated morphology, such as the emergence of binucleated cells, formation of tight gap junctions, and possible bile duct structures. Although OSM alone had very weak effects on hepatocyte functions, albumin secretion and cytochrome P450IA1/2 capacity were greatly enhanced when combined with NA or DMSO. This functional observation closely agreed with the emergence of small hepatocytes. In contrast, ammonium removal was strongly dependent on DMSO alone. DNA amount basis functions of fetal cells with three factors at week 5 were 1/7 for albumin secretion, 3 times higher for ammonium removal, and 1/10 for P450 capacity, compared with those of cultured adult mouse hepatocytes. These results show that inclusion of NA, DMSO, and OSM in the culture medium significantly enhances in vitro maturation of fetal liver cells when compared with conventional culture conditions.

Albumins↗

PubMed2002

Long-term experimental study of the aquatic plant system for polluted river water.

Water purification using artificial wetlands and aquatic macrophyte is attracting attention as a purification technology that can create rich ecosystems while imposing a minimal load on the environment. Because an aquatic plant system requires a large surface area, design specifications and maintenance methods that can obtain the optimum purification effect per unit surface area must be established. Large experimental facilities have been constructed beside a polluted river flowing into Lake Kasumigaura and have been used for a three-year experiment using several kinds of aquatic plants. This report summarizes the characteristics and the design load of the aquatic plant system based on this study and results from other aquatic plant facilities.

Ecosystem↗

PubMed2002

Preservation of human cell bioassay device for the on-site evaluation of environmental waters.

We have already developed a novel disposable bioassay device based on the low-density lipoprotein (LDL) uptaking activity of human hepatoblastoma Hep G2 cells in our previous work. However, this device is not readily applicable to evaluate river water toxicity on-site because it cannot be preserved for more than one week. In this work, we developed the method for preservation of the device to enable it to be preserved for at least one month. The device can be supplied to individual environmental sites without any facilities for cell culture. We can evaluate river water toxicity by 2 hours of exposure after thawing. Therefore, this kind of device could be a promising tool for daily water quality management.

Biological Assay↗

PubMed2001

The membrane-type collectin CL-P1 is a scavenger receptor on vascular endothelial cells.

Collectins are a family of C-type lectins that have collagen-like sequences and carbohydrate recognition domains (CRD). They are involved in host defense through their ability to bind to carbohydrate antigens of microorganisms. The scavenger receptors type A and MARCO are classical type scavenger receptors that have internal collagen-like domains. Here we describe a new scavenger receptor that is a membrane-type collectin from placenta (collectin placenta 1 (CL-P1)), which has a typical collectin collagen-like domain and a CRD. The cDNA has an insert of about 2.2 kilobases coding for a protein containing 742 amino acid residues. The deduced amino acid sequence shows that CL-P1 is a type II membrane protein, has a coiled-coil region, a collagen-like domain, and a CRD. It resembles type A scavenger receptors because the scavenger receptor cysteine-rich domain is replaced by a CRD. Northern analyses, reverse transcription-polymerase chain reaction, and immunohistochemistry show that CL-P1 is expressed in vascular endothelial cells but not in macrophages. By immunoblotting and flow cytometry CL-P1 appears to be a membrane glycoprotein of about 140 kDa in human umbilical vein or arterial endothelial cells, placental membrane extracts, and CL-P1 transfected Chinese hamster ovary cells. We found that CL-P1 can bind and phagocytose not only bacteria (Escherichia coli and Staphylococcus aureus) but also yeast (Saccharomyces cerevisiae). Furthermore, it reacts with oxidized low density lipoprotein (OxLDL) but not with acetylated LDL (AcLDL). These binding activities are inhibited by polyanionic ligands (polyinosinic acid, polyguanylic acid, dextran sulfate) and OxLDL but not by polycationic ligands (polyadenylic acid or polycytidylic acid), LDL, or AcLDL. These results indicate that CL-P1 might play important roles in host defenses that are different from those of soluble collectins in innate immunity.

Amino Acid Sequence↗

PubMed2001

Molecular cloning and functional expression analysis of a cDNA for human hepassocin, a liver-specific protein with hepatocyte mitogenic activity.

By means of differential cDNA expression cloning, we earlier isolated a novel rat cDNA and its protein, named hepassocin, which is upregulated during liver regeneration. Using the rat cDNA as a probe, we have now isolated human hepassocin cDNA encoding a protein of 312 amino acids, which has 81.4% and 83.8% identity, respectively, to rat hepassocin before and after elimination of its signal peptide. Dot blot analysis revealed that hepassocin mRNA was strongly expressed in adult liver, fairly strongly in fetal liver, and weakly in pancreas, but not in other tissues. Recombinant human hepassocin produced in Chinese hamster ovary (CHO) cells by the dihydrofolate reductase-methotrexate (DHFR--MTX) gene amplification method is a homodimer (68 kDa) and has mitogenic activities in hepatocytes of various animal species including rat, mouse, rabbit and dog, and the activity was lost with 2-mercaptoethanol treatment. These results suggest that hepassocin is a potent regulator in liver cell growth not only in rats but also in humans. Computer searches revealed that human hepassocin as well as rat hepassocin has a characteristic disulfide structure close to that of fibrinogen-gamma. We assume that this newly identified growth factor exerts functions in association with an extracellular matrix such as fibrinogen.

Amino Acid Sequence↗

PubMed2001

Involvement of p38 MAP kinase pathway in the synergistic activation of PACAP mRNA expression by NGF and PACAP in PC12h cells.

We have recently shown that in PC12 cells, PACAP and NGF synergistically increase PACAP gene transcription and mRNA level, and that the MAPK/ERK kinase inhibitor PD98059 blocks the PACAP mRNA expression induced by either PACAP or NGF, but not that induced by the combination, suggesting involvement of multiple signaling pathways. Here we show that the p38 MAPK inhibitor SB203580 almost completely inhibits the PACAP mRNA expression induced by PACAP alone or in combination with NGF. PACAP induces neurite outgrowth and potentiates NGF-induced neurite outgrowth in PC12h cells. Unlike the case for the PACAP mRNA expression, SB203580 did not affect, but PD98059 reduced, PACAP and NGF-induced neurite outgrowth. These results indicate that PACAP receptors are coupled to the p38 signaling pathway, and that p38 plays a key role in the regulation of PACAP gene expression, while ERK, but not p38, MAPK is involved in PACAP and NGF-induced neurite outgrowth.

Animals↗

PubMed2001

mRNA sequence of the Xenopus laevis paxillin gene and its expression.

Paxillin is a cytoskeletal protein found in structures of focal adhesions where cells adhere to the extracellular matrix. We isolated paxillin cDNA from the Xenopus laevis ovary. The cDNA sequence encodes a protein of 539 amino acids with four LIM and five LD motifs. 80% of the amino acids of frog paxillin are shared by human and chicken paxillins. Northern analysis showed that the frog gene is expressed in the spleen, kidney, testis and ovary. Immunocytochemistry showed that paxillin protein is accumulated in the nucleus as well as in the periphery of the cytoplasm of the A6 cell. This intriguing result shows that paxillin, which has been characterized as a cytoskeletal protein, is capable of translocating to the nucleus.

Amino Acid Sequence↗

PubMed2001

Human APE2 protein is mostly localized in the nuclei and to some extent in the mitochondria, while nuclear APE2 is partly associated with proliferating cell nuclear antigen.

In human cells APE1 is the major AP endonuclease and it has been reported to have no functional mitochondrial targeting sequence (MTS). We found that APE2 protein possesses a putative MTS. When its N-terminal 15 amino acid residues were fused to the N-terminus of green fluorescent protein and transiently expressed in HeLa cells the fusion protein was localized in the mitochondria. By electron microscopic immunocytochemistry we detected authentic APE2 protein in mitochondria from HeLa cells. Western blotting of the subcellular fraction of HeLa cells revealed most of the APE2 protein to be localized in the nuclei. We found a putative proliferating cell nuclear antigen (PCNA)-binding motif in the C-terminal region of APE2 and showed this motif to be functional by immunoprecipitation and in vitro pull-down binding assays. Laser scanning immunofluorescence microscopy of HeLa cells demonstrated both APE2 and PCNA to form foci in the nucleus and also to be co-localized in some of the foci. The incubation of HeLa cells in HAT medium containing deoxyuridine significantly increased the number of foci in which both molecules were co-localized. Our results suggest that APE2 participates in both nuclear and mitochondrial BER and also that nuclear APE2 functions in the PCNA-dependent BER pathway.

Amino Acid Sequence↗

PubMed2001

The transcription factor FoxH1 (FAST) mediates Nodal signaling during anterior-posterior patterning and node formation in the mouse.

FoxH1 (FAST) is a transcription factor that mediates signaling by transforming growth factor-beta, Activin, and Nodal. The role of FoxH1 in development has now been investigated by the generation and analysis of FoxH1-deficient (FoxH1(-/-)) mice. The FoxH1(-/-) embryos showed various patterning defects that recapitulate most of the defects induced by the loss of Nodal signaling. A substantial proportion of FoxH1(-/-) embryos failed to orient the anterior-posterior (A-P) axis correctly, as do mice lacking Cripto, a coreceptor for Nodal. In less severely affected FoxH1(-/-) embryos, A-P polarity was established, but the primitive streak failed to elongate, resulting in the lack of a definitive node and its derivatives. Heterozygosity for nodal renders the FoxH1(-/-) phenotype more severe, indicative of a genetic interaction between FoxH1 and nodal. The expression of FoxH1 in the primitive endoderm rescued the A-P patterning defects, but not the midline defects, of FoxH1(-/-) mice. These results indicate that a Nodal-FoxH1 signaling pathway plays a central role in A-P patterning and node formation in the mouse.

Animals↗

PubMed2001

GATA DNA-binding protein expressed in mouse I-10 Leydig testicular tumor cells.

A nuclear extract of the mouse I-10 Leydig tumor cell line was analyzed by gel mobility shift assay with a combination of antibodies for various mammalian GATA proteins. Antibodies for GATA-4 caused a super-shift of the DNA-protein complex, which is formed through GATA-4 binding to an oligonucleotide with a typical GATA motif, while ones for GATA-1, GATA-2, GATA-3, and GATA-6 did not. These results indicated that I-10 cells express GATA-4 protein. Western blotting analysis of cellular proteins also demonstrated the presence of GATA-4 protein, the size of which corresponds to that of the rat orthologous protein transiently expressed in Cos-1 cells. A significant level of GATA-4 expression in I-10 cells would be advantageous for studying the roles of this protein, especially in view of gonadal function. We further examined the binding site preference of GATA-4 expressed in I-10 cells. GATA-4 showed broad sequence specificity similar to GATA-6, the order of binding core site preference being GATA > GATT > GATC, and adenine was favored on both sides of the core for strong binding. Thus the conserved zinc finger domain of GATA proteins is suggested to contribute to the binding sequence preference. GATA-4 expressed in I-10 cells was not susceptible to proteolysis coupled with cAMP signaling.

Animals↗