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Biomedical subjects

Y Fukunaga

Publications and source records attributed to Y Fukunaga.

At least 91 records · Page 5Linked to original sources
PubMed1996

Histopathological and immunofluorescent studies on transplacental infection in experimentally induced abortion by equine arteritis virus.

Five pregnant mares, at between 6 and 8 months gestation, were experimentally infected with the Bucyrus strain of equine arteritis virus (EAV). Of the five mares, four aborted and one died. The pathogenesis of the abortions was studied, using histopathologic techniques, tissue immunofluorescence and virus isolation. Common microscopic lesions in the maternal reproductive organs indicated myometritis with a degeneration of the myocytes and an infiltration of the mononuclear cells. Epithelial cells of the endometrial gland showed sporadic degeneration. Lesions in the fetal tissue included an atrophy of the lymphoid follicles in the spleen and lymph nodes with degenerated lymphocytes. The placentae were oedematous and degenerated fibroblasts were observed in the subvillous layers. Immunofluorescence detected EAV antigen in the myometrium and the endometrial gland in the dams, in the subvillous layer of the placentae, and in the aborted fetuses. EAV was recovered from the maternal uteri, placentae and fetuses. The placentae yielded the greatest amounts of the virus. Transplacental infection of the fetus was clearly demonstrated in the EAV infection.

Abortion, Veterinary↗
PubMed1996

Improvement of bone disease with increased dose of glucocerebrosidase in a Gaucher disease patient who had a bone lesion presenting during low-dose enzyme replacement therapy.

In recent years, enzyme replacement therapy has been shown to be useful for the treatment of Gaucher disease. A 10 year old Japanese boy with Gaucher disease underwent splenectomy at the age of 5 years and received enzyme replacement therapy from the age of 6 years. He had avascular necrosis of the bilateral femoral heads, which was not seen at the beginning of the therapy, without deterioration of hematological variables during maintenance therapy. The enzyme dosage was increased from 20 to 120 IU/kg per month resulting in an improvement of the clinical symptoms and bone lesion. In enzyme replacement therapy, dose increase is considered to be essential for improvement in bone disease; however, it is important to watch for the development of bone lesion.

Child↗
PubMed1996

Possible involvement of the total amount of morphine infused in the development of acute morphine dependence in rats.

The severity of naloxone-precipitated withdrawal in rats infused intravenously with morphine at the rates of 2.5, 5 and 10 mg/kg/hr over various time periods was investigated. Plasma morphine concentration reached a constant and rate-dependent level at 1 hr after the start of morphine infusion, and this level was maintained until the termination of infusion. Naloxone (2.0 mg/kg, s.c.) was challenged 18 hr after infusion was stopped, and the withdrawal was evaluated by plasma corticosterone (PCS) increase, diarrhea and body weight loss. The incidence of naloxone-precipitated withdrawal signs was related to both the infusion rate and duration of morphine infusion. The duration of morphine infusion (ET50) needed to elicit naloxone-precipitated PCS increase and diarrhea in 50% of the rats was inversely related to the morphine infusion rates, but the total amount of infused morphine (EA50) that elicited naloxone-precipitated withdrawals in 50% of rats was the same at all infusion rates. These results suggest that the total amount of morphine infused may play an important role in the development of acute physical dependence on morphine rendered by continuous intravenous morphine infusion for 1-8 hr.

Acute Disease↗
PubMed1996

[Flow cytometric measurement of DCFH-oxidation of neutrophils in peripheral whole blood, isolated leukocytes and cerebrospinal fluid cells].

Using 2',7'-dichlorofluorescein diacetate (DCFH-DA) as an indicator for the intracellular formation of H2O2 and free radicals, we measured DCFH-oxidation (DCF fluorescence) of neutrophils by flow cytometry. The DCF fluorescence intensity of unstimulated neutrophils in whole blood was the same as the autofluorescence of neutrophils. However, that of unstimulated isolated neutrophils was higher than autofluorescence. The DCF fluorescence of isolated neutrophils was higher than that of neutrophils in whole blood with or without stimuli, and was decreased to the intensity of neutrophils in whole blood by the addition of plasma or erythrocytes. The DCF fluorescence of neutrophils in whole blood indicates the oxidative state of neutrophils in vivo. The determination of DCFH-oxidation of neutrophils in isolated leukocytes is also necessary to evaluate precisely the oxidative state of neutrophils. In cerebrospinal fluid (CSF) cells from patients with meningitis, the DCF fluorescence intensity of unstimulated neutrophils was higher than the autofluorescence of CSF neutrophils. The DCF fluorescence of CSF neutrophils stimulated with PMA showed increased intensity over that of unstimulated CSF neutrophils. The DCF fluorescence of CSF neutrophils, both unstimulated and stimulated with PMA, were decreased with the addition of plasma but not with the addition of CSF.

Cerebrospinal Fluid↗
PubMed1996

Infantile acute monocytic leukemia with tumor formation in the skin expressing adhesion molecules as seen by electronmicroscopy.

We report a case of infantile acute monocytic leukemia associated with solid tumors in the skin. Light microscopy showed that the tumor cells were mainly spindle-shaped and arranged in tandem. Immunohistochemical staining showed that fibronectin was detected in the extracellular matrix (and partially on the surface of tumor cells), integrin alpha 5 beta 1 on the cell surface, and vinculin and actin were detected in the cytoplasm of the tumor cells. Electron microscopy showed that the tumor cells had perpendicular transmembranous fibrils and closely situated collections of cytoplasmic microfilaments beneath the cell membrane suggesting fibronexus-like structures. We conclude that the expression of fibronectin, integrin alpha 5 beta 1, vinculin, cytoplasmic actin and fibronexus-like structure in leukemic cells indicate these cells possess an adhesive function. The mechanism of formation of the extramedullary solid tumors in this patients involved these adhesion molecules of the tumor cells.

Bone Marrow↗
PubMed1996

[Changes of neopterin in cerebrospinal fluid and serum in children with meningitis].

Neopterin is synthesized mainly by monocytes/macrophages and is considered to be a marker for activation of the cellular immune system. In patients with bacterial or aseptic meningitis, elevated neopterin levels in cerebrospinal fluid (CSF) have been demonstrated. We studied the time courses of CSF and serum neopterin in children with meningitis. The CSF neopterin levels on admission were significantly higher in patients with bacterial meningitis (82.4 +/- 37.0 nmol/L) than in those with aseptic meningitis (32.3 +/- 22.1 nmol/L) or in those with non-pleocytotic CSF (6.9 +/- 4.4 nmol/L). The CSF neopterin levels in the patients with bacterial meningitis were remarkably increased (234.5 +/- 100.2 nmol/L) one day after admission, but the serum neopterin levels were not increased. There was no correlation between CSF neopterin levels and CSF cell count or CSF protein, nor between serum neopterin levels and serum C-reactive protein or peripheral leukocyte count. But the CSF neopterin levels one day after admission were related to the period of positive serum C-reactive protein. CSF neopterin levels in patients with bacterial meningitis were increased one day after admission. The levels in two patients with high levels of CSF IFN-gamma and TNF-alpha were remarkably increased. All patients with bacterial meningitis had received treatment with antibiotics and dexamethasone. It has been reported that TNF-alpha enhances the effect of IFN-gamma for neopterin release by macrophages in vitro and that dexamethasone has the same effect on IFN-gamma as TNF-alpha. The present study suggests that elevation of CSF neopterin in bacterial meningitis results from monocytes/macrophages costimulated with IFN-gamma, TNF-alpha and dexamethasone used in treatment.

Adolescent↗
PubMed1995

Detection of equine arteritis virus (EAV) by polymerase chain reaction (PCR) and differentiation of EAV strains by restriction enzyme analysis of PCR products.

A polymerase chain reaction (PCR) based assay capable of detecting and differentiating seven strains of equine arteritis virus (EAV) from around the world was developed. The primers for the PCR were chosen from the ORF6 gene encoding the unglycosylated membrane protein (M). Viral RNA from cell culture fluids infected with each of the seven EAV strains and RNA from the live vaccine, Arvac, was detected by PCR using four sets of primers. The sensitivity of detection was increased from 100 to 1,000 times by performing nested PCR enabling the detection of RNA at a level of 0.5-5 PFU. Differentiation among the virus strains and the live vaccine was achieved by cutting the PCR-amplified products from three sets of primers with six restriction endonucleases. Using this procedure it was possible to distinguish among the seven EAV strains used.

Base Sequence↗
PubMed1995

Ultrastructure and immuno-cytochemistry of BHK-21 cells infected with a modified Bucyrus strain of equine arteritis virus.

Morphogenesis of a modified Bucyrus strain of equine arteritis virus (EAV) in BHK-21 cells was studied. Bacillary tubules were first detected in the cytoplasm 8 h after infection, and mature virions 79 to 122 nm in diameter, 101 nm on average, were mostly observed in the cisternae of the rough endoplasmic reticulum (RER) at 12 h or later. They had isometrical cores and morphological subunits in the outer layer. Budding occurred from the RER and the outer nuclear membrane, but not from the cell surface. Structural linkage was detected between the tubule and the virus core. Aberrant strands were occasionally demonstrated within the nucleus 12 h after infection, and immunofluorescence and immunogold labeling revealed viral antigen also in the nucleus.

Animals↗
PubMed1995

Seven acylated anthocyanins in the blue flowers of Hyacinthus orientalis.

Two novel anthocyanins, delphinidin 3-O-(6-O-cis-p-coumaroyl-beta-D-glucoside)-5-O-(6-O-malonyl-beta-D -glucoside) and petunidin 3-O-(6-O-trans-p-coumaroyl-beta-D-glucoside)-5-O-(6-O-malonyl-beta -D-glucoside) , were isolated from the blue flowers of Hyacinthus orientalis cv. Delft Blue. Five known acylated anthocyanins, namely, delphinidin 3-O-(6-O-trans-p-coumaroyl-beta-D-glucoside)-5-O-(6-O-beta-D-gluco side), delphinidin 3-O-(6-O-trans-caffeoyl-beta-D-glucoside)-5-O-(6-O-malonyl-beta-D- glucoside), delphinidin 3-O-(6-O-trans-p-coumaroyl-beta-D-glucoside)-5-O-(6-O-malonyl-beta -D-glucoside) , cyanidin 3-O-(6-O-trans-p-coumaroyl-beta-D-glucoside)-5-O-(6-O-malonyl-beta -D-glucoside) and pelargonidin 3-O-(6-O-trans-p-coumaroyl-beta-D-glucoside)-5-O-(6-O-malonyl-beta -D-glucoside) , were also isolated from these flowers. The complete structure of each compound was unambiguously determined by one- and two-dimensional NMR techniques.

Acylation↗
PubMed1995

Ureteral obstruction caused by endometriosis.

The diagnosis of ureteral obstruction caused by endometriosis can be difficult. We present a 51-year-old multiparous Japanese women who had undergone right salpingo-oophorectomy at the age of 19 years. She underwent total hysterectomy, left salpingo-oophorectomy, and adhesiotomy of the right ureter after the diagnosis of myoma uteri and endometriosis was made. Microscopic examination of the surrounding resected tissue confirmed the typical endometriotic origin. In addition to intravenous and retrograde pyelography, computerized tomography was useful to establish the diagnosis of ureteral obstruction caused by an extrinsic mass of endometriotic tissue.

Endometriosis↗
PubMed1995

Crosslines, fluorophores in the AGE-related cross-linked proteins.

We can summarize our results as follows: (1) A pair of fluorescent crosslines were isolated from the Maillard reaction mixture; (2) AGE-proteins contained crossline-like structures, and (3) crossline-like immunoreactivities were accumulated in renal tissues of diabetic rats. From these results we concluded that fluorophores in AGE proteins have crossline-like structures and we had the first indication that XLs could be markers for renal disorders.

Animals↗
PubMed1995

No relation of plasma morphine level to the severity of naloxone-induced withdrawal in acute morphine-dependent rats.

Plasma morphine concentration and naloxone-precipitated withdrawal body weight loss and plasma corticosterone (PCS) increase were determined at 12, 18 and 24 hr after i.v. infusion of morphine at a constant rate of 10 mg/kg/hr for 4 hr in Sprague-Dawley rats. Plasma morphine concentration declined 98.0% within 12 hr and further declined 58.8% during 12-24 hr after morphine infusion. There was a significant difference between plasma morphine concentrations at 12 and 24 hr after the morphine infusion. Naloxone (0.5 and 2.0 mg/kg)-precipitated withdrawal, but not spontaneous withdrawal, was elicited at 12-24 hr after the morphine infusion, and the severity of withdrawal precipitated by 2.0 mg/kg naloxone was the same at 12-24 hr after the morphine infusion. Furthermore, there was no significant correlation between plasma morphine concentration and body weight loss or PCS increase. The results suggest that a constant degree of morphine dependence is sustained during 12-24 hr after the morphine infusion and the severity of naloxone-precipitated withdrawal is not related to the plasma morphine concentration at the time of naloxone injection, that is, the rate of morphine removal from its receptor sites.

Animals↗
PubMed1995

Effects of H-89, an inhibitor of protein kinase A, on the acetylcholine release from myenteric plexus of guinea pig ileum.

In order to clarify the involvement of cyclic AMP-dependent protein kinase (protein kinase A) in acetylcholine (ACh) release from myenteric plexus of guinea pig ileum, the effect of H-89, a specific inhibitor of protein kinase A, on the ACh release was investigated. H-89 (0.1-10 microM) inhibited the spontaneous and nicotine-induced release of ACh in a concentration dependent manner. It at 1 microM decreased both kinds of release of ACh to almost half of the control, but it did not affect the ACh release evoked by electrical field stimulation and by 5-hydroxytryptamine. H-89 had no significant effect on the indomethacin (IND), an inhibitor of PG synthesis, -insensitive component of the spontaneous and nicotine-induced release of, ACh. OP-41483, an analog of PGI2 and forskolin, an activator nicotine-induced release of, ACh. OP-41483, an analog of PGI2 and forskolin, an activator of adenylate cyclase, reversed the inhibitory effect of IND on the ACh release. H-89 at 1 microM completely inhibited the reverse effects of OP-41483 and forskolin. These results suggest that activation of protein kinase A is essential for modulation of the nicotine-induced and spontaneous ACh release from myenteric plexus of guinea pig ileum and the activity of protein kinase A is regulated by endogenous PGs via intracellular cyclic AMP level.

Acetylcholine↗
PubMed1995

Stress-induced cell surface expression and antigenic alteration of the Ro/SSA autoantigen.

Recent studies have shown that Ro/SSA autoantigen is heterogeneous. There are two isoform families; the 60 kD forms (Ro60) and the 52 kD forms (Ro52). Recently we have found that autoantibodies to the Ro/SSA proteins are conformation dependent. Anti-Ro60 antibodies are mainly directed to the native protein and conversely anti-Ro52 antibodies are directed only to the denatured protein. It has been known that UV irradiation to cultured keratinocytes induces cell surface expression of Ro/SSA and this phenomenon has been thought to be related with photosensitivity in patients with anti-Ro/SSA antibodies. We studied the quantitative and qualitative changes of the Ro/SSA protein induced by stress, such as with heat shock and UV irradiation, and found that only Ro52 could be expressed on the cell surface of human peripheral lymphocytes by either heat shock or UV irradiation. Moreover, flow cytometric analysis revealed that HS-treated and UV-treated lymphocytes could be stained with patient sera, and by using a technique which combined immunoprecipitation and Western immunoblotting, it has been confirmed that Ro52 expressed on the cell surface can be recognized by anti-Ro/SSA antibodies in native form while cytoplasmic Ro52 cannot be recognized. These data suggest that Ro52 can be antigenic in vivo when expressed on the cell surface and may explain the mechanism of direct tissue damage by anti-Ro/SSA antibodies.

Autoantibodies↗
PubMed1995

[Sclerotherapy with 3% polidocanol for hydrocele testis].

BACKGROUND: We studied the clinical efficacy of sclerotherapy with injection of 3% polidocanol for hydrocele testis. METHODS: From July, 1992 to March, 1995 sclerotherapy with single injection of polidocanol was performed for 11 patients with 12 hydrocele testis on an outpatient basis. We instilled 3 or 5ml of 3% polidocanol after complete removal of fluid in the hydrocele testis. RESULTS: Complete disappearance on ultrasonography was observed in 75% of the hydrocele testis 6 months after this sclerotherapy. There was neither pain during instillation of 3% polidocanol nor any other complication. Two patients with fluid reaccumulation underwent hydrocelectomy 16 and 6 months after sclerotherapy, respectively. CONCLUSION: This procedure seems to be a safe and useful technique as primary treatment for hydrocele testis.

Adult↗
PubMed1994

Use of the serum neutralisation test for equine viral arteritis with different virus strains.

Serum cross neutralisation tests were conducted with a recent American isolate (84KY-A1) and a European isolate, (Wroclaw-2) and compared with the prototype and modified viruses of the Bucyrus strain of equine arteritis virus by using virus specific immune horse sera. The modified Bucyrus strain was neutralised and showed high neutralisation titres with all the immune sera. The prototype Bucyrus strain was also substantially neutralised, followed by the 84KY-A1 strain. As a result of the tests with the modified Bucyrus strain as the antigen, 20 seropositive horses were discovered among home-bred horses which had no previous record of clinical equine viral arteritis. Heat inactivation of the sera at 62 degrees C for 30 minutes caused the disappearance of all but the most positive reactions. When the prototype Bucyrus strain was used instead of the modified virus, no positive reactions were detectable. A serological survey, using the Bucyrus strain in a microtitre neutralisation test, was conducted with 1656 horse sera collected between 1988 and 1990 in Japan. The test disclosed only eight foreign-bred horses positive to the virus; they had been imported as competition horses. No circumstantial evidence of an effect of such horses as a source of infection for horse populations free of the virus was obtained.

Animals↗