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Biomedical subjects

S Wada

Publications and source records attributed to S Wada.

At least 199 records · Page 11Linked to original sources

Limitations of intraductal ultrasonography in differentiating between bile duct cancer in stage T1 and stage T2: in-vitro and in-vivo studies.

BACKGROUND AND STUDY AIMS: We investigated whether intraductal ultrasonography (IDUS) could distinguish between stage T1 and T2 bile duct cancer. MATERIALS AND METHODS: In-vitro study. Resected bile duct specimens (n = 8) were immersed in a water tank and were pierced with straight pins to clarify the normal layer structure. Ultrasonosgraphic images (20MHz) of the positions of pin echoes were compared to the positions of pin holes as seen on histologic analysis of the specimens. In-vivo study. A thin-caliber high-frequency (6 Fr, 20 MHz) ultrasonic probe was inserted into the bile duct via a transhepatic route or a transpapillary route in 26 patients with bile duct cancer who underwent surgical resection. RESULTS: In-vitro study. The inner hypoechoic layer on the IDUS image corresponded not only to the fibromuscular layer but also to a part of fibrous layer of the perimuscular loose connective tissue on histologic analysis, especially in the cases with moderate to severe bile duct wall fibrosis. The outer hyperechoic layer corresponded to the subserosal fat tissue. In-vivo study. In four of six patients with tumor limited to the inside hypoechoic layer on IDUS images, the histologic findings showed tumor invasion to the fibrous layer of the perimuscular loose connective tissue. Due to this limitation, accuracy of IDUS in T-staging was only 20/26 (77 %). CONCLUSIONS: IDUS cannot reliably distinguish bile duct cancer in stage T1 from that in stage T2.

Adult↗

Macrophage migration inhibitory factor in the human ovary: presence in the follicular fluids and production by granulosa cells.

Cytokines play an important role in ovarian function. We unexpectedly found high expression of macrophage migration inhibitory factor (MIF) mRNA in human ovarian tissues. Hence, we examined the presence of MIF in the follicular fluid because the follicular microenvironment is important for oocyte fecundity. The follicular fluids were collected from ovaries of patients undergoing in vitro fertilization and embryo transfer. A higher amount of MIF was identified in the follicular fluid, 80.3 +/- 4.6 ng/ml (mean +/- SE), in which the concentration was significantly decreased as the size of the follicles increased. To detect MIF mRNA expression in the granulosa cells, reverse transcription-polymerase chain reaction was carried out and this showed an amplified transcript specific for MIF. Furthermore, the presence of MIF protein in the granulosa cells was confirmed by Western blot analysis. These results suggest the possibility that MIF mediates various immunological events in the process of oocyte development.

Cells, Cultured↗

Plant 21D7 protein, a nuclear antigen associated with cell division, is a component of the 26S proteasome.

Previously, we cloned a carrot (Daucus carota L.) cDNA encoding a 45-kD protein, 21D7, located in the nuclei of proliferating cells. The 21D7 protein is similar to the partial sequence of a regulatory subunit of the bovine 26S proteasome, p58 (G. DeMartino, C.R. Moomaw, O.P. Zagnitko, R.J. Proske, M. Chu-Ping, S.J. Afendis, J.C. Swaffield, C.A. Slaughter [1994] J Biol Chem 269: 20878-20884) and to the deduced sequence encoded by the Saccharomyces cerevisiae gene SUN2 (M. Kawamura, K. Kominami, J. Takeuchi, A. Toh-e [1996] Mol Gen Genet 251: [146-152]). In our work, the expression of plant 21D7 cDNA rescued the yeast sun2 mutant. Fractionation of carrot and spinach (Spinacia oleracea L.) crude extracts showed that the 21D7 protein sedimented with the active 26S proteasomes. The cessation of cell proliferation in carrot suspensions at the stationary phase caused 26S proteasome dissociation and, correspondingly, the 21D7 protein sedimented together with the free regulatory complexes of the 26s proteasomes. Large-scale purification of carrot 26s proteasomes resulted in co-isolation of the 21D7 protein. Polyacrylamide gel electrophoresis under nondenaturing conditions showed that the 21D7 protein had the same mobility as the 26S proteasome and that proteasome dissociation changed the mobility of the 21D7 protein accordingly. We conclude that the 21D7 protein is a subunit of the plant 26S proteasome and that it probably belongs to the proteasome regulatory complex.

Animals↗

Regulation by calcitonin and glucocorticoids of calcitonin receptor gene expression in mouse osteoclasts.

We previously studied regulation of the calcitonin (CT) receptor (CTR) by glucocorticoid (GC) and CT in cultures of mature mouse osteoclast-like cells (OCLs). The present studies were designed to examine the interaction of CT and GC in regulation of the CTR in osteoclasts and the molecular mechanisms involved. Treatment of OCLs with 10(-7) M dexamethasone (Dex) increased the CTR number in a time-dependent manner, whereas treatment with 10(-9) M salmon CT (sCT) reduced CTR number; neither treatment changed receptor affinity. Dex pretreatment somewhat antagonized the CT-induced reduction in [125I]sCT specific binding. Dex increased, and sCT pretreatment decreased, the sCT-responsive adenylate cyclase activity in parallel with the change in receptor binding. Dex treatment resulted in an increase in CTR messenger RNA (mRNA) levels, as assessed by reverse transcription-PCR, indicating that the increased CTR number was mediated by de novo CTR synthesis. This effect was specific to GCs and was not reproduced by mineralocorticoids or sex steroids. Treatment with sCT resulted in a rapid and profound reduction in CTR mRNA expression, and this reductions was somewhat delayed by Dex pretreatment. OCLs were treated with 5,6-dichloro-1 beta-D-ribofuranosyl benzimidazole to enable estimation of the mRNA decay rates in the absence of ongoing transcription. The stability of CTR mRNA was similar to the control value in Dex-treated OCLs, suggesting that the effect of Dex may be due to changes in transcriptional activity. Interestingly, transcriptional inhibition by 5,6-dichloro-1 beta-D-ribofuranosyl benzimidazole abolished the ability of CT to reduce CTR mRNA levels, suggesting that CT may act by increasing the rate of CTR mRNA decay, and that this effect requires ongoing transcription. The 3'-untranslated region of the mouse CTR mRNA contains four copies of the AUUUA motif, as well as other A/U-rich sequences, which have been shown to determine the stability of other mRNA transcripts. The stability results were consistent with the results of the nuclear transcript run-on assay, which indicated that treatment with Dex enhanced the rate of transcription, whereas CT had no effect. These results show that GC and CT influence CTR expression by distinct mechanisms and provide the basis for identification of the cellular factors involved.

Animals↗

Color Doppler ultrasonography in the diagnosis of portal vein invasion in patients with pancreatic cancer.

We retrospectively evaluated the diagnostic usefulness of color Doppler ultrasonography for detecting portal vein invasion in 21 patients with pancreatic cancer who underwent surgical exploration (14 resection, seven inspection). Real-time gray scale ultrasonography, color Doppler ultrasonography, computed tomography, and angiography were performed in all patients to evaluate portal vein invasion, and the images were compared with the histopathologic or surgical inspection findings. On comparison between gray scale ultrasonographic and color Doppler ultrasonographic images, the tumor-vessel relations were visualized more clearly on color Doppler ultrasonography than on gray scale ultrasonography in 14 (22.2%) of 63 vessels. The sensitivity of color Doppler ultrasonography, computed tomography, and angiography for diagnosing portal invasion was 73.7%, 73.7%, and 73.6%, and the specificity was 95.1%, 95.5%, and 90.9%, respectively; the overall accuracy was 84.1%, 88.9% and 85.7%, respectively. A mosaic signal pattern was observed in 12 vessels and showed an accuracy of 86.4% for diagnosing portal vein invasion. In conclusion, compared with gray scale ultrasonography, color Doppler ultrasonography provided improved images of the tumor and portal vein. Furthermore, the accuracy of color Doppler evaluation of portal vein invasion appears to be equal to that of computed tomography and angiography. Therefore, color Doppler ultrasonography may play an important role as an initial examination for evaluating portal vein invasion.

Adult↗

Changes in lymphocyte subsets following administration of interleukin 2 and cyclophosphamide in mice with transitional cell carcinoma.

We investigated the effects of combination therapy with interleukin 2(IL-2) and cyclophosphamide (CPM) in C3H/HeN mice implanted with mouse bladder tumor cells (MBT2). Tumor growth was suppressed in the mice treated with IL-2 alone and mice treated with CPM alone, although not completely. However, tumor growth was completely suppressed in the mice treated with both IL-2 and CPM. As for lymphocyte subset analysis of the spleen using flow cytometry, the frequency of Lyt2+ cells and asialo GM1+ cells was significantly higher in the mice treated with IL-2 and CPM compared to the other groups. In the lymphocyte subset analysis of the thymus, the frequency of L3T4-Lyt2+ cells was significantly lower in the mice treated with CPM alone and mice treated with IL-2 and CPM compared to the other groups.

Animals↗

Leuconostoc bacteremia after liver transplantation: another cause of vancomycin resistant gram-positive infection.

Leuconostoc sp. are gram-positive bacteria intrinsically resistant to vancomycin, which can be confused with streptococci based on routine microbiological-characteristics. Infections secondary to Leuconostoc are uncommon, and usually affect patients with underlying diseases, prior use of vancomycin and those with central lines. The most common clinical presentation is fever secondary to a central line infection. We report the first case of Leuconostoc infection in a solid organ transplant recipient. The patient developed Leuconostoc bacteremia secondary to peritonitis, 60 d after undergoing liver transplantation. He was treated with clindamycin, gentamicin and underwent surgical debridement, but succumbed to other complications.

Adult↗

A case of membranous glomerulonephritis associated with gastric cancer.

We describe a patient with gastric cancer and membranous glomerulonephritis (MGN). The patient, a 61-year-old male, was admitted to our Hospital in May, 1996, because of proteinuria and hyperlipidemia persisting for a year. Laboratory examination filled the criteria of nephrotic syndrome and renal biopsy revealed MGN of stage II. Prednisolone therapy (40 mg/day p.o.) was started, followed by a gradual decrease in proteinuria from 4.5 g/day to 0.1 g/day. Endoscopic examination was performed because of stomach-ache revealed advanced gastric cancer of Borrmann 4. Desiring for a conservative therapy, he was discharged and moved to a hospice. In literature review, MGN is the most frequent lesion among various glomerular diseases associated with malignancy, such as the lung, stomach, and colon, particularly at an elderly ages, and sometimes antedates the detection of malignancy, as in the present case. In several cases with MGN, immune-complexes composed of tumor antigens, such as carcino-embryonic antigen, and antibodies have been reported to deposit in basement membrane of glomeruli, causing MGN. In the renal and gastric cancer tissues of the present case, the presence of three novel tumor-associated antigens, Span-1, Thomsen-Friedenreich antigen (T antigen) and F1 alpha antigen, was examined, using a immuno-peroxidase method. Although none of these three antigens were immuno-stained in the renal tissue, clinical course and literature review suggest that MGN in this patient seems to be associated with gastric cancer, which may have produced MGN-causing tumor antigens other than the three antigens. It should be emphasized that malignancy should be carefully and routinely examined in patients with MGN, particularly at elderly ages.

Glomerulonephritis, Membranous↗

Beneficial effects of perfluorotributylamine/pluronic F-68 stem-emulsion (FC43se) on discordant lung xenografts.

The aim of this study was to assess the effectiveness of Perfluorotributylamine/Pluronic F-68 Stem-Emulsion (FC43se) against hyperacute rejection in rabbit-pig xenodiscordant transplantation in an ex-vivo lung model. Rabbits were divided into two groups. In Group 1 (control), after extraction, the lungs were connected directly to the experimental circuit as soon as possible. The lungs were then perfused with 100 ml of pig blood only. In Group 2 (FC(+) Group), after extraction, the lungs were connected to the same circuit as soon as possible. The lungs were then perfused with 10 ml of FC43se plus 90 ml of pig blood. The duration of perfusion was defined from when the perfusion started until the PAP reached 50 mmHg. Significant differences in survival were seen between Group 1 and Group 2: in Group 1, the survival time was 51.9 +/- 16.8 min, whereas in Group 2 the survival time was 76.9 +/- 12.4 min (p < 0.01). The wet-to-dry weight ratio after 30 min of perfusion in Group 2 was significantly lower than that of Group 1 and the mean pulmonary artery pressure of Group 2 at 35, 40 and 45 min after perfusion was significantly lower than that of Group 1. Histological examination revealed that FC43se suppressed the adhesion of leukocytes to the surfaces of endothelial cells, and also attenuated the intimal edematous changes accompanying leukocyte infiltration. Therefore, FC43se has a beneficial effect on suppressing hyperacute rejection in xenogeneic discordant lung transplantation.

Animals↗

Carnosine sustains the retention of cell morphology in continuous fibroblast culture subjected to nutritional insult.

L- Carnosine (beta-alanyl L-histidine), occurring abundantly in skeletal muscles, has been suggested to possess antioxidant and anti-aging properties. Using three different experimental approaches (microscopic, flow cytometric and ELISA for one of the markers of DNA oxidative damage) this study on rat embryonic fibroblasts demonstrates that L-carnosine at 30 mM concentration sustains the retention of cell morphology even during a nutritional insult for five weeks. Also, L-carnosine significantly reduces the formation of 8-hydroxy deoxyguanosine (8-OH dG) in the cells after four weeks of continuous culture. Thus it could be inferred that the anti-senescent effect of L-carnosine is probably linked to its inhibition of formation of intracellular 8-OH dG during oxidative stress.

8-Hydroxy-2'-Deoxyguanosine↗

Introduction of basic fibroblast growth factor gene into mouse renal cell carcinoma cell line enhances its metastatic potential.

To clarify the role of basic fibroblast growth factor (FGF-2) in the malignant progression of renal cell carcinoma, we transfected the FGF-2 gene, which lacks the typical signal sequence, into RenCa, a mouse renal cell carcinoma cell line that does not express FGF-2 mRNA. In an in vitro tumor cell invasion assay, the FGF-2-transfected cell lines (RenCa/F) exhibited 3- to 4-fold higher invasive potential than either the parental RenCa (RenCa/P) or the vector-only transfected cell line (RenCa/C). Zymography showed a marked increase in matrix metalloproteinase 2 (MMP-2) production in the culture supernatants of RenCa/F. Furthermore, when injected i.v. or into the renal subcapsule in syngeneic mice, RenCa/F formed more than 10 times as many metastatic nodules in the lung as did RenCa/P and RenCa/C. Metastases to the liver and mesenteric lymph nodes were observed only after the injection of RenCa/F into the renal subcapsule. In contrast, there was no significant difference in either cell proliferation in vitro or tumor growth in vivo among RenCa sublines. These results suggest that if it is overexpressed, endogenous native FGF-2 plays an important role in the invasion and metastasis of renal cell carcinoma, probably through the production of MMP-2.

Animals↗

Fungal metabolites. XXI. Characteristics of low energy collision induced dissociation of [M + 2H]2+, [M + H + Na]2+ and [M + 2Na]2+ of peptaibols using electrospray ionization mass spectrometry.

The mass spectral characteristics of selected peptaibols were investigated using electrospray ionization in conjunction with low-energy collision-induced dissociation (CID). Protonated and sodiated molecular species were selected as precursor ions and the resulting CID spectra are discussed with respect to fragmentation characteristics related to the structures of peptaibols. The CID spectra of peptaibols with acetylated N-terminus and with the sub-sequences of Aib-Pro were similar. The CID spectra of [M + 2Na]2+ provided structural information more than those of [M + 2H]2+ and [M + H + Na]2+. The CID of [M + 2Na]2+ can be used to deduce complete sequence information for peptaibols.

Amino Acid Sequence↗

Pancreatic arteriovenous malformation with duodenal ulcer. Demonstration by color Doppler ultrasonography.

We report the color Doppler ultrasonography features of arteriovenous malformation (AVM) of the pancreas, a very rare disease. The patient was a 52-year-old man with congenital AVM of the pancreas and a duodenal ulcer that had been resistant to medication. Endoscopic color Doppler ultrasonography (color Doppler EUS) revealed many abnormal color signals showing pulsatile wave form at the portion of the duodenal wall involving the duodenal ulcer. Extracorporeal color Doppler ultrasonography revealed a mosaic-like color signal, caused by turbulent flow, in the portal trunk. Angiography demonstrated a vascular network with extensive proliferation at the pancreatic head and early portal filling. It is possible that the pancreatic AVM had caused the duodenal ulcer. Color Doppler EUS can be a useful modality for detection of vessel abnormalities of the gastrointestinal tract.

Arteriovenous Malformations↗

Hereditary ceruloplasmin deficiency with hemosiderosis.

Hereditary ceruloplasmin deficiency with hemosiderosis (aceruloplasminemia) is a new disease characterized by systemic hemosiderosis, diabetes mellitus, neurological abnormalities and pigment degeneration of the retina. Loss of the ferroxidase activity of ceruloplasmin results in systemic iron deposition and tissue damage. Neuroimaging studies reveal iron deposition in basal ganglia and in the red and dentate nuclei. Cerebellar ataxia, extrapyramidal signs and dementia develop after middle age. We report a patient with undetectable serum ceruloplasmin levels and the above clinical manifestations. Sequence analysis of the cDNA of ceruloplasmin from this patient revealed an insertion of adenine in exon 3; this produced a premature stop codon.

Amino Acid Sequence↗

Urinary type IV collagen as a marker for early diabetic nephropathy.

We investigated the urinary secretion of type IV collagen in 115 subjects with non-insulin-dependent diabetes mellitus (NIDDM) without macroproteinuria, 34 normal healthy subjects and 19 subjects with chronic glomerulonephritis (CGN). We examined the relation between the urinary level of type IV collagen and various clinical parameters. The urinary level of type IV collagen was significantly elevated in NIDDM subjects compared with normal subjects (4.88 +/- 3.12 vs. 1.7 +/- 1.25 micrograms/gCr, P < 0.001). The urinary level of type IV collagen was increased even in NIDDM subjects with normoalbuminuria. The ratio of urinary type IV collagen was significantly lower in subjects with chronic glomerulonephritis (CGN) than those in NIDDM subjects (P < 0.001), although there was no significant difference in the urinary level of type IV collagen between NIDDM and CGN subjects. The ratio of urinary type IV collagen to albumin was under 10.0 x 10(-6) in all subjects with CGN. Our results suggest that measurement of the urinary level of type IV collagen is useful for detection of early diabetic nephropathy and for the differential diagnosis of diabetic nephropathy and chronic glomerulonephritis.

Adult↗

c-fos antisense DNA inhibits proliferation of osteoclast progenitors in osteoclast development but not macrophage differentiation in vitro.

We previously reported that osteoclast formation in vitro, by coculture of mouse bone marrow and primary osteoblastic cells, occurs in two phases: proliferation of osteoclast progenitors followed by terminal differentiation into mature osteoclasts. Using this coculture system, we examined the effects of c-fos antisense and sense phosphorothioate oligonucleotides on osteoclast development and macrophage differentiation. Treatment with c-fos antisense for the first 4 days of coculture inhibited osteoclast formation in a dose-dependent fashion. However, when c-fos antisense was added during the second phase of coculture (4-6 days), osteoclast formation was unaffected. In contrast, c-fos antisense treatment had no effect on the appearance of F4/80 antigen-positive cells of the macrophage lineage in these cultures or on the induction by colony stimulating factor-1 of macrophage colony formation in cultures of mouse bone marrow cells in agar. Neither osteoclast differentiation nor macrophage appearance was inhibited by adding control c-fos sense in the cocultures. When c-fos antisense was added into an assay of bone resorption by mature osteoclasts, pit formation on dentine slices was unaffected. These results indicate that c-fos plays an important role in the proliferative phase of osteoclast progenitors in osteoclast development, but not in the terminal differentiation phase or in the bone resorbing activity of mature osteoclasts. c-fos antisense specifically inhibited osteoclast formation but had no effect on macrophage development.

Analysis of Variance↗