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Biomedical subjects

Y Fukui

Publications and source records attributed to Y Fukui.

At least 73 records · Page 4Linked to original sources

Prenatal exposure to ethanol induces leptomeningeal heterotopia in the cerebral cortex of the rat fetus.

Pregnant rats were fed an ethanol-containing liquid diet between gestational day (GD) 10 and GD 21. Leptomeningeal heterotopias were observed in the cerebral cortex of ethanol-exposed fetuses. They appeared on the brain surface of the lateral cortical region near the rhinal fissure, and were found more numerously in the rostral than the caudal region. These abnormalities contained certain neuronal perikarya, microtubule-associated protein (MAP) 1b-positive neuronal processes, and Rat-401-positive radial glial fibers. Immunostaining for Rat-401 revealed that the heterotopias protruded through breaches in the glia limitans. In adult rats exposed to ethanol prenatally, the heterotopias persisted in the lateral cortical region. We conclude that prenatal exposure to ethanol might induce defects in the glia limitans, resulting in the genesis of leptomeningeal heterotopias. These abnormalities may be related to mental retardation or the cognitive deficits associated with human fetal alcohol syndrome (FAS).

Alcohol-Induced Disorders, Nervous System↗

Abnormalities of cerebellar foliation in rats prenatally exposed to ethanol.

Pregnant rats were given a liquid diet containing 5% (w/v) ethanol between gestational days 10 and 21. Cerebella of their offspring were examined at 7 weeks of age. Rats exposed prenatally to ethanol showed a fusion of folia V and VI in the cerebellar vermis. Around the fusion, the cortical laminar structure was disrupted: Purkinje cell dendrites derived from each adjacent folium were tangled, and solitary or clustered ectopic granule cells were in the molecular layer. Some ectopic granule cells surrounded basophilic rosette-like structures. Glial fibrillary acidic protein immunostaining revealed defects in the glia limitans, which is formed by Bergmann glial endfeet on the cerebellar surface. Absence of the glia limitans was observed corresponding to the fusion area. These findings suggested that prenatal exposure to ethanol might impair the formation of the glia limitans in the cerebellum, resulting in the fusion of folia and the disruption of the cortical structure. These malformations may be involved in the delayed motor development and ataxia seen in human fetal alcohol syndrome.

Abnormalities, Drug-Induced↗

An increased expression of Ca(2+) channel alpha(1A) subunit immunoreactivity in deep cerebellar neurons of rolling mouse Nagoya.

Rolling mouse Nagoya (RMN) is an ataxic mutant and carries a mutation in the gene coding for the alpha(1A) subunit of the P/Q-type Ca(2+) channel. We examined the immunohistochemical expression of the alpha(1A) subunit in deep cerebellar nuclei of RMN. The antibody used recognized residues 865-883 of the mouse alpha(1A) subunit not overlapping the altered sequences in RMN. In RMN, many neurons exhibited definite alpha(1A) subunit-staining in the medial nucleus, interposed nucleus, and lateral nucleus of deep cerebellar nuclei. The number of positive neurons in these nuclei was significantly higher in RMN than in controls. Increased expression of the alpha(1A) subunit in deep cerebellar neurons might compensate for the altered function of the P/Q-type Ca(2+) channel of RMN.

Animals↗

Topological relationship between corticotropin-releasing factor-immunoreactive cerebellar afferents and tyrosine hydroxylase-immunoreactive Purkinje cells in a hereditary ataxic mutant, rolling mouse Nagoya.

Using immunohistochemistry we examined the distribution of corticotropin-releasing factor-positive cerebellar afferents and the topological relationship between their projections and the distribution of tyrosine hydroxylase-positive Purkinje cells in an ataxic mutant, rolling mouse Nagoya. In the mutants, some climbing fibers were more intensely stained for corticotropin-releasing factor, but their zonal distribution remained the same as in non-ataxic littermates (control mice). These climbing fibers arose from the dorsal accessory nucleus, the ventral lamella of principal nucleus, the dorsomedial cell group, the subnucleus A, the beta subnucleus and the ventrolateral protrusion of the inferior olive, since perikarya in these olivary subdivisions were more intensely stained for corticotropin-releasing factor than in controls. Some mossy fiber rosettes in the vermal lobules, the simple lobule, the crus I of ansiform lobule, the copula pyramidis and the flocculus also exhibited corticotropin-releasing factor immunoreactivity and were more densely stained in the mutants than in controls. Double immunostaining for corticotropin-releasing factor and tyrosine hydroxylase in the mutant cerebellum revealed that the distribution of tyrosine hydroxylase-positive Purkinje cells corresponded to terminal fields of corticotropin-releasing factor-positive climbing fibers but not corticotropin-releasing factor-positive mossy fibers. This study indicated an increased corticotropin-releasing factor immunoreactivity in some climbing or mossy fibers in the cerebellum of rolling mouse Nagoya. We also found that the distribution of tyrosine hydroxylase-positive Purkinje cells corresponded to terminal fields of corticotropin-releasing factor-positive climbing fibers in the mutant cerebellum. As the transcription of the tyrosine hydroxylase gene is facilitated by Ca2+, abnormal tyrosine hydroxylase expression in the mutant Purkinje cells may indicate functional abnormality by alterations in intracellular Ca2+ concentrations. Therefore, we suggest that an increased level of corticotropin-releasing factor in a specific population of climbing fibers may alter the function of their target Purkinje cells.

Animals↗

An attempt at intracytoplasmic sperm injection of frozen-thawed minke whale (Balaenoptera bonaerensis) oocytes.

Little is known about the characteristics of fertilisation events in minke whales. Cryopreserved minke whale oocytes and spermatozoa do not fertilise in a standard IVF. This study was conducted to investigate the pronucleus formation ability of cryopreserved minke whale oocytes and their subsequent development following intracytoplasmic sperm injection (ICSI). In experiment 1, frozen-thawed minke whale immature oocytes were cultured for in vitro maturation (IVM) in a maturation medium (TCM199) supplemented with either porcine follicle stimulating hormone (pFSH)/estradiol-17beta (E2) or pregnant mare's serum gonadotropin (PMSG)/human chorionic gonadotropin (hCG). After 120 h of IVM, oocyte survival was examined before ICSI, and showed no significant difference in morphological normality (24-36%) between the two IVM media. Two-cell embryos (two oocytes from 21 sperm-injected oocytes) were obtained when the maturation medium was supplemented with pFSH/E2 or PMSG/hCG. In experiment 2, cryopreserved maturing oocytes were investigated for the effects of repeat-culture (2 h or 24 h) on survival before ICSI. Pronuclear formation and development were examined for the effects of sperm pretreatment with dithiothreitol (DTT) and oocyte activation with ethanol at ICSI. A frequency of 49-69% of frozen-thawed maturing oocytes was used for ICSI. Although oocyte activation did not produce a significant difference in survival, pronucleus formation and embryonic development, 2- and 4-cell cleaved oocytes were observed after injection of sperm pretreated with DTT.

Animals↗

Renal arteriovenous fistula developing after tumor enucleation using a microwave tissue coagulator.

With the increase in detection of incidental renal cell carcinoma, nephron-sparing surgery for small renal cell carcinomas is now recognized as one of the surgical options. We report a case of renal arteriovenous fistula developing after non-ischemic tumor enucleation of a small renal cell carcinoma using a microwave tissue coagulator. A 50-year-old Japanese man presented with right flank pain and gross hematuria. The patient had undergone non-ischemic tumor enucleation for right renal cell carcinoma, 2 cm in diameter, 1 month previously. Doppler ultrasound revealed the formation of an arteriovenous fistula at the enucleated portion. Transcatheter super-selective occlusion of the feeding artery was successfully performed with two metallic coils. The patient has been followed up with no sign of recanalization of the fistula. In this case, the tumor was located close to the renal hilus with thick arterial branches around the tumor. Additional microwave coagulations against arterial bleeding from the cutting surface might have been the cause of the fistula formation of this case. Non-ischemic tumor enucleation using a microwave tissue coagulator is a relatively easy and secure nephron-sparing surgical procedure. Excessive coagulation, however, should be avoided, since it might be the cause of unexpected postoperative vascular complications.

Arteriovenous Fistula↗

Estimation of left ventricular recovery level based on the motor current waveform analysis on circulatory support with centrifugal blood pump.

In a mock circulatory loop simulating the left heart bypass using a centrifugal blood pump, analysis of the motor current waveform of the centrifugal pump was performed to derive a useful parameter to evaluate the status of ventricular function. The relationship between the peak, amplitude, and the peak of the fundamental frequency of the power spectral density of the periodic motor current waveform (MCpsdP) that reflected the pulsatile ventricular pressure, and the peak of the left ventricular pressure (LVP) was examined. Although both peak and amplitude of the motor current waveform showed an excellent correlation with the peak LVP, they failed to predict the opening of the aortic valve. The MCpsdP that corresponds to the frequency of the heart rate showed an excellent correlation with the peak LVP throughout the LVP levels, but the slope between them changed with the opening of the aortic valve. Thus, it is possible to follow the change in the LVP and detect even the opening of the aortic valve, and, hence, the recovery of the left ventricle. However, the slope of the linear regression equation varied, depending on the pump speed. This result implies that the MCpsdP can be possibly used to follow the change of ventricular function during circulatory assistance with a centrifugal blood pump as well as to control the pump speed in response to varying ventricular function.

Centrifugation↗

Ethanol and oxidative stress.

This article represents the proceedings of a workshop at the 2000 ISBRA Meeting in Yokohama, Japan. The chair was Albert Y. Sun. The presentations were (1) Ethanol-inducible cytochrome P-4502E1 in alcoholic liver disease, by Magnus Ingelman-Sundberg and Etienne Neve; (2) Regulation of NF-kappaB by ethanol, by H. Matsumoto, Y. Nishitani, Y. Minowa, and Y. Fukui; (3) Chronic ethanol consumption increases concentration of oxidized proteins in rat liver, by Shannon M. Bailey, Vinood B. Patel, and Carol C. Cunningham; (4) Antiphospholipids antibodies and oxidized modified low-density lipoprotein in chronic alcoholic patients, by Tomas Zima, Lenka Fialova, Ludmila Mikulikova, Ptr Popov, Ivan Malbohan, Marta Janebova, and Karel Nespor; and (5) Amelioration of ethanol-induced damage by polyphenols, by Albert Y. Sun and Grace Y. Sun.

Alcoholism↗

Traumatic subluxation of the axis after hyperflexion injury of the cervical spine in children.

Six cases of children (four boys and two girls, mean age 11 years) who had traumatic subluxation of the axis (C2) were reviewed retrospectively. Initial radiographs demonstrated no detectable vertebral fracture in any of the children. However, a slight anterior subluxation of C2 was observed in three of the patients. Radiographs, taken at 1 month after injury in all but one patient, revealed a progression of the subluxation and a local kyphosis in all of the patients. Four of the children were treated conservatively with a cervical brace, and an improvement of both the kyphosis and the anterior slippage of C2 was obtained accompanied by an anteroposterior growth of the C3 vertebral body. The kyphosis of two of the patients became severe and, ultimately, these patients underwent fusion surgery. At the follow-up, none of the patients presented with any significant symptom. For the correct diagnosis of traumatic subluxation of C2, sequential radiographs to confirm the progression of subluxation and local kyphosis are mandatory. Conservative treatment rather than early surgical treatment may be chosen for this injury, because mild and moderate kyphosis can be corrected spontaneously by remodeling of the cervical spine.

Adolescent↗

Organ-specific autoimmunity in mice whose T cell repertoire is shaped by a single antigenic peptide.

Organ-specific autoimmune diseases have been postulated to be the result of T cell response against organ-specific self-peptides bound to MHC molecules. Contrary to this paradigm, we report here that transgenic mice lacking MHC class I expression and expressing an MHC class II I-A(b) molecule that presents only a single peptide (E alpha 52-68) spontaneously develops peripheral nervous system-specific autoimmune disease with many of the histopathological features found in experimental allergic neuritis. Reciprocal bone marrow chimeras produced using susceptible and resistant lines revealed that bone marrow-derived cells determined disease susceptibility. While the expression of the I-A(b)-E alpha 52-68 complex in the periphery was readily detectable in both lines, its expression on thymic dendritic cells responsible for tolerance induction was markedly lower in the susceptible line than in the resistant line. Consistent with this, CD4(+) T cells that can be activated by the I-A(b)-E alpha 52-68 complex were found in the susceptible line, but not in the resistant line. Such CD4(+) T cells conferred the disease to the resistant line by adoptive transfer, and administration of Ab specific for the I-A(b)-E alpha 52-68 complex inhibited disease manifestation in the susceptible line. These results indicate that disease development involves systemic T cell reactivity to I-A(b)-E alpha 52-68 complex, probably caused by incomplete negative thymocyte selection.

Animals↗

First cutaneous branch of the internal pudendal artery: an anatomical basis for the so-called gluteal fold flap.

We investigated the cutaneous blood supply in the gluteal and perineal regions of 35 donated cadavers to provide an anatomical basis for reliable vulvo-vaginal reconstruction using a skin flap such as the so-called gluteal fold flap. The cutaneous areas along the gluteal cleft and sulcus were likely to be supplied by 3 routes: 1) the internal pudendal artery (IPA), especially its first cutaneous branch; 2) perforators running through the gluteus maximus muscle and arising from the inferior gluteal artery (IGA); and 3) a non-perforator running around and inferior to the ischial tuberosity and originating from the IGA. Route 1 supplied the skin along the gluteal cleft, route 2 the gluteal fold (i.e., a bulky skin fold along the upper edge of the gluteal sulcus), and route 3, just along the gluteal sulcus. In those 3 routes, we noted the consistent morphology of the thick and long, first cutaneous branch of the IPA. The first arterial branch, 1.5 mm in diameter at its origin on average (ranging from 0.7-2.6 mm), usually originated from the IPA under the cover of or at the inferomedial or distal side of the sacrotuberous ligament (almost always less than 20 mm from the inferomedial margin of the ligament). The branch ran superomedially toward the coccyx or ran medially in the ischiorectal fat. It accompanied the vein and nerve at its distal (peripheral) course although the nerve often ran independently at its proxomal course near the ligament. Therefore, the first branch of the IPA seems to provide a reliable pedicle using the skin along the gluteal cleft whether the incision for approach is conducted along the gluteal sulcus or not. However, if the gluteus maximus muscle extended much inferomedially, the pedicle would be very short. In this case, preparation of the pedicle seems to be necessary along the arterial course under the cover of the muscle.

Aged↗

Push-out strength of hydroxyapatite coated by sputtering technique in bone.

Hydroxyapatite was coated with 1 microm thickness on titanium columns of a length of 10 mm, an outer diameter of 4.0 mm by radio frequency magnetron sputtering. The hydroxyapatite coating titanium columns were implanted in the diaphysis of femora of 3 adult dogs, and push-out test was carried out after 2, 4 and 12 weeks of implantation using a testing machine. The interface of bone/column was observed histologically after the test. At 12 weeks the push-out strengths of coating and non-coating columns were 3.5 and 1 MPa, respectively. Histological observation indicated a formation of thin connective tissue with 5-30 microm thickness at the interface of the bone/column. No inflammation was observed during the implantation periods.

Animals↗

Expression of tyrosine hydroxylase in cerebellar Purkinje cells of ataxic mutant mice: its relation to the onset and/or development of ataxia.

This report describes recent studies on tyrosine hydroxylase (TH) expression in Purkinje cells of the cerebellum of ataxic mutant mice. An increased expression of TH in some Purkinje cells has been observed in two allelic groups of mutant mice, tottering and dilute. TH-positive Purkinje cells appeared preceding the onset of ataxia. Northern blot analysis revealed 2.1 kb of TH mRNA in the mutant cerebella, and the size was identical to that of TH transcripts in other brain regions. However, TH in Purkinje cells did not seem to participate in catecholamine biosynthesis. In vitro studies showed that cultured non-catecholaminergic neurons expressed the TH transcripts following Ca2+ influx. Therefore, abnormal TH expression in the mutant Purkinje cells may indicate neuronal dysfunction caused by misregulation of intracellular Ca2+ concentrations.

Animals↗

Lipid and fatty acid analysis of fresh and frozen-thawed immature and in vitro matured bovine oocytes.

The lipid content and fatty acid composition of fresh immature and in vitro matured bovine oocytes cultured in media with or without serum, and also those of frozen-thawed immature oocytes were analysed. All oocytes were ranked (A or B) on the basis of their cytoplasmic quality. Fatty acid composition (mol %; w/w) in the total lipid fraction was analysed by gas chromatography. Triglyceride, total cholesterol, phospholipid (phosphocholine-containing phospholipid) and non-esterified fatty acid contents of immature and in vitro matured oocytes were determined using lipid analysis kits. Phosphocholine-containing phospholipid and non-esterified fatty acid contents were determined in frozen-thawed immature bovine oocytes. Palmitic acid was the most abundant fatty acid in immature oocytes (A: 35%, B: 36%), and in in vitro matured oocytes cultured in the medium containing serum (A: 36%, B: 35%) or polyvinyl alcohol (A: 33%, B: 36%). Oleic acid was the second most abundant fatty acid in all A ranked oocytes, whereas stearic acid was the second most abundant fatty acid in all B ranked oocytes. There were significant differences (P < 0.05) in linoleic and arachidonic acid fractions between A and B ranked immature oocytes. In vitro matured oocytes had significantly (P < 0.05) lower proportions of linoleic and arachidonic acids, and significantly (P < 0.01) lower contents of triglyceride and total cholesterol compared with those of immature oocytes. The fatty acid composition of in vitro matured oocytes cultured in medium containing fetal calf serum or polyvinyl alcohol was similar, but significant differences (P < 0.01) in triglyceride and the total cholesterol content were observed. There was a significant decrease (P < 0.05) in the arachidonic acid proportion in frozen-thawed immature oocytes compared with that in fresh immature oocytes. In addition, significant (P < 0.05) decreases in both phospholipid (15.8--10.6 pmol) and non-esterified fatty acid (11.0--4.1 pmol) were found in frozen--thawed immature oocytes. The results indicate that lipids are available for use as an energy source for maturation and that serum lipids are incorporated into the oocyte cytoplasm during in vitro maturation. The changes in the lipid content (mainly phospholipid) and fatty acid composition were also observed in frozen--thawed immature oocytes. The study indicates that the alteration of fatty acid composition in bovine oocytes might improve maturation and cryopreservation.

Animals↗

[Screening methods in the detection of bladder cancer: comparison of nuclear matrix protein-22, bladder tumor antigen and cytological examinations].

We evaluated the utility of urinary parameters (Nuclear Matrix Protein-22: NMP-22, Bladder Tumor Antigen: BTA, and cytological examinations) for the diagnosis or post-therapeutic monitoring of bladder cancer. Thirty one tumor-bearing cases including 19 fresh cases and 40 tumor-free cases, were subjects of this study. Using identical voided urine samples, NMP-22, BTA and urinary cytology were examined. The mean values of NMP-22 (cut-off value is 12 U/ml) was 100.5 +/- 26.5 U/ml in the tumor-bearing group and 21.9 +/- 7.8 U/ml in the tumor-free group (p < 0.05): Sensitivity was 74.2%, and specificity was 67.5%. Sensitivity of BTA was 58.1%, and specificity was 97.5%. Only five cases were judged positive by urinary cytology: 16.1% in sensitivity and 100% in specificity. Thus, NMP-22 and BTA were more sensitive than urinary cytology. In conclusion, the new urinary parameters, NMP-22 and BTA, would be less invasive and useful as tumor markers of bladder cancer. NMP-22 seems suitable for screening before the diagnosis and BTA for the post-therapeutic follow-up study.

Adult↗

Novel role of phosphatidylinositol 3-kinase in CD28-mediated costimulation.

Ligation of the CD28 surface receptor provides a major costimulatory signal for full scale T cell activation. Despite extensive studies, the intracellular signaling pathways delivered by CD28 ligation are not fully understood. A particularly controversial matter is the role of phosphatidylinositol 3-kinase (PI3K) in CD28-mediated costimulation. It is known that the binding site for PI3K and Grb-2 lies nested within the YMNM motif of the CD28 cytoplasmic domain. To elucidate the role of PI3K during CD28-mediated interleukin-2 (IL-2) production, CD28 YMNM point and deletion mutants were expressed in Jurkat cells. We then measured IL-2 promoter activation after CD28 ligation. Our results showed that the Y189F mutant, which disrupts binding by PI3K, and the YMNM deletion mutant both demonstrated reduced but significant activity for IL-2 promoter activation. In contrast, the N191A mutant, which retains PI3K binding ability, resulted in a complete abrogation of activity, suggesting that PI3K mediates a negative effect upon transcriptional activation of the IL-2 gene. Consistent with this idea, we found that the addition of a PI3K pharmacological inhibitor augmented IL-2 promoter activity, whereas coexpression of a constitutively active form of PI3K reduced this activity. Taken together, these data indicate that PI3K, when associated with the YMNM motif, may act as a negative mediator in CD28-mediated IL-2 gene transcription.

Amino Acid Motifs↗

Diversity of T cell repertoire shaped by a single peptide ligand is critically affected by its amino acid residue at a T cell receptor contact.

T cell differentiation in the thymus is driven by positive selection through the interaction of alphabeta T cell receptors (TCRs) with self-peptides bound to self-major histocompatibility complex molecules, yet the influence of the peptide sequence on this process remains unknown. To address this issue, we have compared CD4(+) T cell differentiation between two sets of mouse lines in which MHC class II I-A(b) molecules are occupied with either Ealpha chain-derived peptide ((p)Ealpha) or its variant, (p)60K, with one amino acid substitution from leucine to lysine at P5 residue of TCR contacts. Here, we show that despite the comparable expression of I-A(b)-peptide complex in the thymus, this substitution from leucine to lysine affects efficiency of positive selection, resulting in extremely small numbers of CD4(+) T cells to be selected to mature on I-A(b)-(p)60K complex. Furthermore, we show that, although I-A(b)-(p)Ealpha complex selects diverse T cells, T cell repertoire shaped by I-A(b)-(p)60K complex is markedly constrained. Our findings thus suggest that positive selection is both specific and degenerate, depending on the amino acid residues at TCR contacts of the selecting self-peptides.

Amino Acid Sequence↗