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Biomedical subjects

K Fukuda

Publications and source records attributed to K Fukuda.

At least 469 records · Page 26Linked to original sources

Ossification of the posterior longitudinal ligament. Autosomal recessive trait.

STUDY DESIGN: This study analyzed the mode of inheritance of ossification of the posterior longitudinal ligament (OPLL) from the pedigree of a family. OBJECTIVES: The results were correlated to provide a new mode of inheritance of OPLL. SUMMARY OF BACKGROUND DATA: Although a nation-wide multicenter survey of OPLL has been carried out in 347 subjects and 1030 relatives in Japan since 1981, no parental consanguinity has been reported. METHODS: In the family, three siblings and one other member of a family underwent operations for OPLL in our department, and the clinical information regarding other family members was obtained from interviews with these four patients or relatives. RESULTS: The parents of three affected siblings and another unaffected sister were first cousins, and the father was suspected to be affected as well. CONCLUSION: Transmission of OPLL as an autosomal recessive trait in this family, which has not been reported, is suspected; although, the possibility that it is a dominant trait can not be excluded.

Aged↗

Risk factors for hepatocellular carcinoma among Japanese women.

To elucidate the risk factors for hepatocellular carcinoma (HCC) among women, we made a combined analysis of the data from three case-control studies conducted in high-risk areas of Japan. A total of 120 cases and 257 controls were included in the analysis. After adjustment for the study category, age, and other potential confounders, significantly increased risks were associated with chronic hepatitis-B virus infection (odds ratio [OR] = 42.4, 95 percent confidence interval [CI] = 11.2-160.2), a past history of blood transfusion (OR = 3.7, CI = 1.8-7.5), and a history of smoking (OR = 2.2, CI = 12-4.1). In addition, women with a history of heavy drinking experienced an elevated risk of borderline significance (OR = 4.2, CI = 0.9-20.4, P = 0.07). When these ORs were compared with the corresponding estimates among males from the same case-control studies, no significant differences were observed between the two genders. Among the factors examined in this analysis, drinking and smoking habits--which are more common among Japanese men than women--may partly account for a large male-predominance in the incidence of HCC. Further studies are needed to clarify the roles that sex-hormones and hepatitis-C virus infection might play in the large gender difference of HCC occurrence.

Adult↗

Migration of the yeast linear DNA plasmid from the cytoplasm into the nucleus in Saccharomyces cerevisiae.

The Kluyveromyces linear plasmids, pGKL1 and pGKL2, carrying terminal protein (TP), are located in the cytoplasm and have a unique gene expression system with the plasmid-specific promoter element termed UCS, which functions only in the cytoplasm. In this study we have developed an in vivo assay system in Saccharomyces cerevisiae which enables the detection of a rare migration of the yeast cytoplasmic plasmid to the nucleus, using a pGKL1-derived cytoplasmic linear plasmid pCLU1. pCLU1 had both the UCS-fused LEU2 gene (a cytoplasmic marker) and the native URA3 gene (a nuclear marker) and therefore its cytoplasmic-nucleo localized could be determined by the phenotypic analysis of the marker. The nuclearly migrated plasmids were often detected as linear plasmids having the telomere sequence of the host yeast at both ends, although circular plasmids were also found. The circular form was produced by the the terminal fusion of pCLU1. Insertion of a Ty element into a nuclearly migrated plasmid was observed, allowing the ROAM-regulated expression of the adjacent nuclearly silent UCS-fused LEU2 gene. The nuclearly located plasmids, whether linear or circular, were less sensitive to UV-mediated curing than pGKL and pCLU1.

Base Sequence↗

Inositol 1,4,5-trisphosphate formation and ryanodine-sensitive oscillations of cytosolic free Ca2+ concentrations in neuroblastoma x fibroblast hybrid NL308 cells expressing m2 and m4 muscarinic acetylcholine receptor subtypes.

Intracellular free Ca2+ concentrations ([Ca2+]i) were measured in subclones of NL308 neuroblastoma x fibroblast hybrid cells expressing each of the individual muscarinic acetylcholine receptor (mAChR) subtypes m1, m2, m3 and m4. Application of 100 microM acetylcholine (ACh) increased [Ca2+]i in all four subclones. The increased [Ca2+]i levels were significantly higher in m1- and m3-transformed cells than those in m2- and m4-transformed cells. In more than 95% of m2- and m4-transformed cells, [Ca2+]i showed sinusoidal oscillations. ACh-induced increases in [Ca2+]i were not observed in cells treated with an intracellular Ca2+ chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). Removal of extracellular Ca2+ with ethylene-glycol-bis-(beta- aminoethyl)-N,N,N',N'-tetraacetate (EGTA) did not affect the initial [Ca2+]i increases, but reduced the late phases of delta [Ca2+]i in ml- and m3-transformed cells by 20-30%. Oscillations in m2- and m4-transformed cells persisted in EGTA solution (though sometimes slowed in frequency), suggesting that they were of intracellular origin. ACh-induced delta [Ca2+]i and inositol 1,4,5-trisphosphate formation was completely suppressed by pre-treatment with 50-100 ng ml-1 Pertussis toxin (PTX) for 12 h in m2- and m4-transformed cells, but not in m1- and m3-transformed cells. In all cells, extracellular application of caffeine and ryanodine, or intracellular application of cyclic adenosine diphosphate ribose (cAD-PR) produced a rise in [Ca2+]i. ACh-induced [Ca2+]i oscillations were not observed in ryanodine-treated m2-transformed cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗

Zonal differences in nitric oxide synthesis by bovine chondrocytes exposed to interleukin-1.

To determine the role of nitric oxide (NO) in the inhibition of aggrecan synthesis, we measured levels of NO produced by bovine chondrocytes from different layers of articular cartilage in the presence of interleukin-1 (IL-1). Chondrocytes from the superficial layer showed a large increase in NO synthesis in response to IL-1. Although chondrocytes from the deep layer also produced NO in response to IL-1, the amount was less than that from the superficial layer. Enhanced NO production evoked by IL-1 was accompanied by a significant inhibition of aggrecan synthesis. These data suggest that chondrocytes in both superficial and deep layer of articular cartilage inhibit aggrecan synthesis with IL-1 via NO production. In addition, superficial layer cells respond to lower amounts of IL-1 with respect to NO-production and inhibition of proteoglycan synthesis.

Animals↗

Interleukin-1 inhibits keratan sulfate production by rabbit chondrocytes: possible role of prostaglandin E2.

We investigated the importance of prostaglandin E2 (PGE2) release in interleukin-1 (IL-1)-induced inhibition of aggrecan synthesis by chondrocytes. Keratan sulfate (KS) production was measured in parallel with PGE2 release in chondrocytes. IL-1 inhibited KS production and stimulated PGE2 release. In the presence of PGE2, there was a dose-dependent decrease in baseline KS production. Indomethacin and dexamethasone partially blocked the IL-1-induced PGE2 release while KS production recovered. Our results suggest that IL-1 inhibits KS production, in part, by stimulating the release of PGE2.

Animals↗

Opioid mu- and kappa-receptor mediate phospholipase C activation through Gi1 in Xenopus oocytes.

In the Xenopus oocytes expressing mu- or kappa-opioid receptors, agonist-induced currents were observed only when the oocyte was coinjected with Gi1 alpha RNA and pretreated with K-252a, a potent inhibitor of protein kinases. The evoked currents were abolished by intracellular injection of EGTA or inositol 1,4,5-trisphosphate and the current-voltage relationship revealed that they are mediated through typical calcium-dependent chloride channels. These findings suggest that the mu- and kappa-receptors mediate phospholipase C activation through Gi1 alpha, and that these receptor mechanisms including downstream signalings might be inhibited by phosphorylation in vivo in the Xenopus oocyte.

Animals↗

Superficial repositioning of the artery for chronic hemodialysis: indications and prognosis.

Superficial repositioning of the artery (SRA) is a modality of the blood access operation for chronic hemodialysis that has been previously used in cases of cardiac failure. We performed 42 SRAs from 1986 to 1993; thereafter, we retrospectively investigated the operative indications, postoperative complications, and long-term results. Superficial repositioning of the artery was indicated for the lack of an appropriate vein (17 cases; 40%), frequent and early access failure due to arteriovenous fistula or polytetrafluoroethylene grafts (six cases; 14%), venous hypertension (five cases; 12%), and cardiac failure (two cases; 5%). The patency rates of the SRAs were 87% at 3 years and 58% at 4.5 years. There was some difficulty in finding the returning veins in five of 28 functioning SRAs (18%). The SRA is thus considered to be a secondary-selected blood access operation; however, it also may be used as an efficient blood access for an extended period of time without any serious complications.

Adult↗

Effects of oxygen radicals on ciliary motility in cultured human respiratory epithelial cells.

There are few reports about direct effects of specific oxygen products on ciliary function because of their instability and reactivity. We investigated the direct effects of superoxide anion (O2-) and of hydrogen peroxide (H2O2) on the ciliary function of human respiratory epithelial cells, using monolayer cell cultures, high speed video analysis of frequency (CBF), amplitude (CBA), and coordination of ciliary beats and evaluating the surface structural changes of ciliated cells at the same time. 10(-2) M H2O2 decreased ciliary beat activity. The CBF was 36.5 +/- 4.4% and the CBA was 51.0 +/- 3.8% of the baseline (time = 0) after 5 min (all p < 0.001). Catalase (2 micrograms/ml) abolished the ciliotoxic effect of H2O2. The O2- produced by reaction of xanthine (0.06 mM)-xanthine oxidase (0.04 U/ml) caused a temporary rapid increase of 26.8 +/- 1.7% in CBF and an increase of 42.5 +/- 4.1% in CBA after 15 sec (all p < 0.001). Superoxide dismutase significantly reduced these increases. Results indicated that O2- activated ciliary function with a temporary increase in O2(-)-production. This suggests that the removal of H2O2 from the O2- reaction is important in improving mucociliary clearance in excessive oxygen metabolites.

Cells, Cultured↗

Coexpression of cdk2/cdc2 and retinoblastoma gene products in colorectal cancer.

The retinoblastoma gene (Rb gene) is a tumour-suppressor gene and its product (pRB) is known to act as a negative regulator of the cell cycle. Although lack of pRB expression resulting from gene alterations is considered to be responsible for the genesis of several human malignancies, increased expression of pRB has been demonstrated in a majority of colorectal cancer cases. In the present study, we investigated the expression of pRB as well as that of its related kinases, cdk2 and cdc2, in colorectal cancer, since these kinases have been reported to phosphorylate and inactivate pRB. Western blot analysis revealed that colorectal cancer expressed higher levels of cdk2 and cdc2 than did normal mucosa and that the ratio of the hyperphosphorylated form of pRB was higher in colorectal cancer. Furthermore, immunohistochemical studies showed that cdk2/cdc2 was expressed exclusively in the cancer cells positive for pRB. These results suggest that an increase in the expression of cdk2/cdc2 in colorectal cancer may have prevented pRB from braking the cell cycle through phosphorylation.

Blotting, Western↗

Coupling of the cloned mu-opioid receptor with the omega-conotoxin-sensitive Ca2+ current in NG108-15 cells.

Voltage-dependent Ca2+ currents were measured in NG108-15 neuroblastoma x glioma hybrid cells transformed to express the rat mu-opioid receptor by the whole-cell configuration of the patch-clamp technique with Ba2+ as charge carrier. A mu-opioid receptor-selective agonist, [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin caused significant inhibition of voltage-dependent Ca2+ currents in mu-receptor-transformed NG108-15 cells but not in nontransfected or vector-transformed control cells. On the other hand, a delta-opioid receptor-selective agonist, [D-penicillamine2,D-penicillamine5]enkephalin, induced inhibition of voltage-dependent Ca2+ currents in both control and mu-receptor-transformed cells, which is mediated by the delta-opioid receptor expressed endogenously in NG108-15 cells. The inhibition of voltage-dependent Ca2+ currents induced by [D-Ala2,N-Me-Phe4,Gly5-ol]enkephalin [D-penicillamine2,D-penicillamine5]enkephalin was reduced by pretreatment of the cells with pertussis toxin or omega-contoxin GVIA. These results indicate that the mu-opioid receptor expressed from cDNA functionally couples with omega-contoxin-sensitive N-type Ca2+ channels through the action of pertussis toxin-sensitive G proteins in NG108-15 cells.

Animals↗

Immunolocalization of endothelin in the traumatized spinal cord: relationship to blood-spinal cord barrier breakdown.

The purpose of this study was to determine the relationship between endothelin-1 (ET-1), a prominent vasoactive agent, and the breakdown of the blood-spinal cord barrier along the axis of the cord after a moderate spinal cord injury. In the first study rats (n = 10) were euthanized 24 h after spinal cord injury and compared to sham (n = 5) and unoperated (n = 10) controls. Endothelin and immunoglobulins (IgG) were immunolocalized in adjacent sections of spinal cord using semiquantitative immunocytochemical techniques. In the second study animals were pretreated with the endothelin antagonist Bosentan (n = 6) or vehicle (n = 6) prior to spinal cord injury. Animals were euthanized at 24 h postinjury. Ten minutes prior to euthanasia animals were given horseradish peroxidase (HRP) intravenously. After perfusion fixation sections of cord were prepared for quantitative HRP histochemistry. After spinal cord injury there was enhanced staining for endothelin along the axis of the cord that correlated with the anatomical pattern of barrier breakdown to IgG. In those animals that were pretreated with Bosentan, there was a significant reduction in barrier breakdown along the axis of the injured cord as compared to those animals that received vehicle only. Taken together, this data implicate involvement of endothelin in the axial pattern of barrier breakdown after spinal cord injury.

Animals↗

The blood-brain barrier disruption to circulating proteins in the early period after fluid percussion brain injury in rats.

Breakdown of the blood-brain barrier (BBB) immediately after traumatic brain injury is not clearly understood. In the present study we focused on the integrity of the BBB to circulating proteins within the first hour after injury. For this purpose, vascular permeability to endogenous albumin and to the exogenous protein tracer horseradish peroxidase (HRP) was examined after a lateral fluid percussion brain injury in rats. Albumin was immunolocalized in brain sections at 3 and 60 min after impact. This distribution was compared with the histochemical localization of HRP given before impact at the same time points. In a separate experiment HRP was given prior to sacrifice to determine the time course for the barrier disruption. Permeability to this protein was assessed at 13, 30, and 60 min after impact. Prominent extravasation of albumin occurred within 3 min of injury and was present in multiple foci within the injured hemisphere. At 60 min the extravasated albumin was present in the same sites, where it was widely distributed. Throughout the related brain parenchyma, little difference was found between the extravascular distribution of albumin and HRP. In the delayed administration paradigm breakdown of the BBB was noted in the impact site, hemorrhagic site in the deep cortical layer, hippocampus, thalamus, and midbrain at 13 min after injury. This injured barrier was restored in most regions by 30 min. However, the impact site and hemorrhagic site remained permeable up to 60 min postinjury. In addition, newly developed barrier disruption to HRP occurred in the parasagittal cortex at 30 and 60 min. In conclusion, widespread breakdown of the BBB to circulating proteins occurred within a few minutes after traumatic brain injury. The time course for this barrier disruption is characterized by three different patterns: (1) transient, (2) prolonged, and (3) delayed opening. This variation in the development of barrier disruption may be related to the secondary barrier failure as well as the primary opening after injury.

Animals↗

Altered immunoexpression of microglia and macrophages after mild head injury.

In this study we examined the temporal response of microglia and macrophages to mild head injury in the rat. Microglia and macrophages were identified by their distinct morphology and by immunophenotype. With regard to the latter, antibodies to OX42 and ED1 were used to define microglia and macrophages, respectively. Although there was no change in the morphology of brain macrophages after mild head injury, the morphology of microglia was dramatically altered. Microglial cell bodies appeared larger with a more elaborate arborization of cellular processes. After head injury certain populations of macrophages and microglia were more intensely immunostained. By 3 days postinjury these intensely stained cells exhibited a characteristic distribution in the brain. Prominently stained microglia were detected in the thalamus, hippocampus, lateral and medial geniculate body, and the substantia nigra. Intensely stained macrophages were located primarily in the cortex and subarachnoid space adjacent to the site of impact. By 7 days postinjury intensely immunostained macrophages and microglia were widespread throughout the injured cortex. These results demonstrate that microglia and macrophages are sensitive to mild head injury. Early changes in the macrophage population are more directly correlated with the most damaged tissue and may reflect migration of these cells from either the subarachnoid space or across the damaged blood-brain barrier. The early widespread microglial response in regions exhibiting no overt neuronal cell damage suggests that these cells are responding to more subtle factor(s) that are expressed in the mildly traumatized brain.

Animals↗

Evidence for sexual and mother-to-child transmission of human T lymphotropic virus type II among Guaymi Indians, Panama.

Guaymi Indians, a non-intravenous drug-using population in which human T cell lymphotropic virus type II (HTLV-II) is endemic, were studied in Changuinola, Panama, to identify the prevalence and modes of transmission of HTLV-II. A population-based survey showed that 352 (9.5%) of the 3686 participants were seropositive for HTLV-II. Infection rates were the same for male and female subjects and increased significantly with age, beginning in young adulthood. HTLV-II infection status was highly concordant among spouses (P < .001) and between mother and child; of children aged 1-10 years, 36 of 219 born to seropositive mothers were seropositive compared with 3 of 997 born to seronegative mothers (P < .001). The strong associations of HTLV-II infection with age and with an infected spouse in adults and of infection in children with infection in their mothers strongly suggest sexual and mother-to-child transmission of HTLV-II in this population.

Adolescent↗

Healthy and atretic follicles: vaginosonographic detection and follicular fluid hormone profiles.

The health status of human preovulatory follicles was prospectively studied by non-invasive transvaginal ultrasound. Daily ultrasound scans were performed from the time when the follicle measured 15 mm in diameter until formation of corpus luteum or oocyte retrieval. Based on the ultrasound scans two types of follicles were defined: type A follicles showed a cloud visualized as a cone projecting into the follicular fluid with the base of the cone positioned on the follicle wall. A light spot seen at the tip of the cloud in the majority of scans was presumed to be the oocyte-cumulus complex. Type B follicles showed an echo free space without a cloud. Two groups of infertility patients were studied: Group I received intrauterine insemination; 106 patients undergoing a total of 263 cycles (188 spontaneous cycles and 75 clomiphene citrate cycles). Group II received in-vitro fertilization (IVF); 22 patients undergoing a total of 52 cycles (31 spontaneous cycles and 21 clomiphene cycles). In the first group, the ovulatory potential of the follicle is associated with the ultrasound characteristics. From the IVF patients the follicular fluid was harvested and assessed for the free concentration of the following steroids: oestradiol, progesterone, testosterone and androstenedione. The endocrine health status of the follicle was associated with the ultrasound characteristics of the follicle. In patients of group I, 288 type A follicles ovulated out of a total of 298 follicles (97%). None of the type B follicles ovulated (0/14). In patients of group II, oocytes were obtained in 79% of the type A follicles (50/63), whereas 7% of the type B follicles yielded an oocyte (1/14).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗