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Biomedical subjects

K Fukada

Publications and source records attributed to K Fukada.

At least 37 records · Page 2Linked to original sources
PubMed1997

Immunohistochemistry of collagen types II and X, and enzyme-histochemistry of alkaline phosphatase in the developing condylar cartilage of the fetal mouse mandible.

We investigated the immunohistochemical localisation of types II and X collagen as well as the cytochemical localisation of alkaline phosphatase in the developing condylar cartilage of the fetal mouse mandible on d 14-16 of pregnancy. On d 14 of pregnancy, although no immunostaining for types II and X collagen was observed, alkaline phosphatase activity was detected in all cells in the anlage of the future condylar process. On d 15 of pregnancy, immunostaining for both collagen types was simultaneously detected in the primarily formed condylar cartilage. Alkaline phosphatase activity was also detected in chondrocytes at this stage. By d 16 of pregnancy, the hypertrophic cell zone rapidly increased in size. These findings strongly support a periosteal origin for the condylar cartilage of the fetal mouse mandible, and show that progenitor cells for condylar cartilage rapidly or directly differentiate into hypertrophic chondrocytes.

Alkaline Phosphatase↗
PubMed1997

Tissue-specific and ontogenetic regulation of LIF protein levels determined by quantitative enzyme immunoassay.

To define the physiological role of leukemia inhibitory factor (LIF), it is essential to localize sites of LIF synthesis in vivo. We generated polyclonal antibodies specific for native rat LIF, and developed a two-site immunoassay to detect 10 pg LIF/ml. Using this immunoassay, we determined LIF content of 18 organs, CNS regions, and ganglia throughout postnatal development of rats. High levels of LIF protein (1.0-11.0 ng/g tissue) are present in relatively few tissues: the uterus at late proestrus to estrus and on day 5 of pregnancy, ovary at estrus to early metestrus-1, footpads during early postnatal development and thymus throughout. Intermediate levels (0.5-1.0 ng) are detected in the gut, skin, skeletal muscle, pancreas and lung at one or more postnatal ages. Low levels (0.1-0.5 ng) are observed in most other non-nervous and nervous tissues. LIF protein levels do not completely correspond to reported LIF mRNA levels.

Age Factors↗
PubMed1994

Cholinergic neuronal differentiation factor (CDF)/leukemia inhibitory factor (LIF) binds to specific regions of the developing nervous system in vivo.

The cholinergic neuronal differentiation factor (CDF)/leukemia inhibitory factor (LIF) promotes survival and/or differentiation of specific neuronal populations in vitro. To investigate the spatial and temporal distribution of CDF/LIF binding sites in the developing rat nervous system, we localized binding of iodinated native rat CDF/LIF by radioautography. We find that specific CDF/LIF binding sites are present in distinct regions of the nervous system at defined developmental stages. CDF/LIF binding is detectable in all sensory ganglia examined, both neural crest- and placode-derived, and in all sympathetic and parasympathetic ganglia examined. In the central nervous system, in contrast, binding is restricted to specific regions in the motor, sensory, and limbic systems. These in vivo results not only support culture studies which suggest CDF/LIF involvement in development of specific populations of neurons, but also suggest new roles played by CDF/LIF.

Animals↗
PubMed1994

Mutations in the transcriptional regulatory region of the precore and core/pregenome of a hepatitis B virus with defective HBeAg production.

Termination mutations in the precore open reading frame of hepatitis B virus (HBV) variants with defective hepatitis B e antigen (HBeAg) production have been demonstrated in both infected patients who have seroconverted to anti-HBe and those with fulminant hepatitis B. A donated plasma sample was found to be positive for the hepatitis B surface antigen, but negative for both HBeAg and anti-HBe. The HBV DNA titer in the plasma was estimated to be 32 pg/ml, and circulating virus-like particles were observed by electron microscopy. The entire nucleotide sequence of the virus was determined and at least 7 nucleotides were found to be unique when compared with previously reported sequences. These nucleotides created no termination codon in the precore/core, pol, preS/S and HBx open reading frames. The deduced amino acid substitutions were 28 Arg--Gln, 94 His--Tyr, 131 Val--Ile and 132 Phe--Tyr of HBx and 715 Met--Val and 789 Asp--Asn of pol. Furthermore, the precore and core/pregenome promoter contained altered 1764 A, 1766 T and 1768 A. Therefore, mutations in regions other than the precore open reading frame can cause defective HBeAg production.

Adult↗
PubMed1993

Mother-to-child transmission of human T-lymphotropic virus type I (HTLV-I): an extended follow-up study on children between 18 and 22-24 years old in Okinawa, Japan.

The significant difference observed between the seroprevalence of HTLV-I in adults and in children is as yet unexplained. To evaluate a hypothetical explanation of the existence of seroconversion cases of "seronegative carriers" for this phenomenon, 21 of 55 children who had been born to seropositive mothers and who remained seronegative until the age of 18 years were further followed up at the ages of 22 and/or 24 years. None of the 21 seronegative children born to seropositive mothers seroconverted, either at 22 years or at 24 years. In addition, the polymerase-chain-reaction (PCR) technique could not prove the existence of the HTLV-I provirus genome in peripheral mononuclear cells (PBMC) of 10 of these children. Our results fail to prove the possibility of viral latency of HTLV-I in mother-to-child transmission. Therefore, the hypothetical seroconversion of "seronegative carriers" after adulthood cannot be an explanation.

Adolescent↗
PubMed1993

Detection of antibodies to hepatitis C virus (HCV) structural proteins in anti-HCV-positive sera by an enzyme-linked immunosorbent assay using synthetic peptides as antigens.

We have defined 10 linear immunogenic regions encoded by the putative hepatitis C virus (HCV) structural proteins (core and envelope) by employing an enzyme-linked immunosorbent assay (ELISA) and by using 17 sequential synthetic peptides covering the N-terminal 330 amino acids of the structural polyproteins as antigens. These peptides correspond to amino acids 1 to 24, 21 to 44, 42 to 68, 64 to 91, and 100 to 120 of the putative core protein and amino acids 192 to 212, 223 to 238, 236 to 258, 250 to 266, and 307 to 330 of the putative envelope protein. In particular, the peptide covering amino acids 21 to 44 of the core protein was reactive with all but one (40 of 41) of the serum samples giving a positive signal in the passive hemagglutination assay (PHA) using the core and nonstructural proteins (NS 3/4) of the virus as antigens. We detected the HCV genome in 25 (61%) of 41 PHA-positive serum samples by the polymerase chain reaction (PCR) test. Of 25 PCR-positive serum samples, 17 serum samples had reactivity to the peptides derived from the envelope protein. On the other hand, only 1 of the 16 PCR-negative serum samples had reactivity to the peptides derived from the envelope protein. Interestingly, we often observed high serum alanine aminotransferase levels in PCR-positive individuals bearing antibodies to the envelope protein.

Alanine Transaminase↗
PubMed1992

Development of a supersensitive polymerase chain reaction method for human T lymphotropic virus type II (HTLV-II) and detection of HTLV-II proviral DNA from blood donors in Japan.

A supersensitive polymerase chain reaction procedure was developed to detect human T-lymphotropic virus type II (HTLV-II) proviral genome. Six primer pairs covering the various regions of HTLV-II were compared and selected on the basis of specificity and sensitivity. Among them, one primer pair of the pol region of HTLV-II (II pol) was able to amplify and detect even 0.1 fg of the cloned plasmid HTLV-II DNA (seven copies) by regular ethidium bromide staining on polyacrylamide gel. By using this procedure, we screened 189 HTLV-I seropositive blood donors from Yamaguchi and Fukuoka Red Cross Blood Centers, Japan. There were four positive samples detectable with the HTLV-II-specific pol primer pair, as well as with the HTLV-I tax primer pair. The amplified DNAs of two specimens were cloned and sequenced. The sequences of the HTLV-I tax region from both specimens were identical to that of HTLV-I. On the other hand, those of the HTLV-II pol region were identical to that of HTLV-II, except for one base substitution in a clone from one subject. These results indicate that dual infection of HTLV-I and HTLV-II in the same persons occurs among Japanese blood donors.

Base Sequence↗
PubMed1992

Some chemical properties of the HCl-methanol extract from the puparial cuticle of Drosophila melanogaster.

1. The HCl-methanol (HCl-MeOH) soluble fraction from the puparial cuticle of yellow, black and ebony of D. melanogaster was hydrolyzed in hydrochloric acid and examined for beta-alanine, ketocatechol, and acetic acid. 2. Between beta-alanine and ketocatechol and between beta-alanine and acetic acid, a quantitatively inverse relationship was found, respectively. The former relationship was further confirmed by the feeding experiment of beta-alanine to black. 3. Of total beta-alanine in the HCl-MeOH extract, the proportion of those having free amino group was 74.8 per cent. 4. All these results indicate that the HCl-MeOH soluble fraction of the puparial cuticle may be useful for investigating the cross-link structure of the cuticle.

Acetates↗
PubMed1992

Function-blocking antibodies against cholinergic neuronal differentiation factor.

Cholinergic neuronal differentiation factor, CDF, causes a transition from noradrenergic to cholinergic phenotype in cultured sympathetic neurons. Moreover, its identification with leukemia inhibitory factor has shown that CDF is a multifunctional cytokine. To examine the physiological role of CDF and to further elucidate the as yet unknown effects of CDF on the nervous system, two kinds of function-blocking antibodies were generated. One type, raised against whole native CDF, completely blocks CDF activity, whereas the other type, raised against a synthetic peptide corresponding to the N-terminal amino acid region of CDF, blocks activity partially. All three anti-CDF and two antipeptide polyclonal antibodies tested in this study significantly inhibit CDF function.

Animals↗
PubMed1991

Immunoaffinity purification and dose-response of cholinergic neuronal differentiation factor.

A glycoprotein from heart cell-conditioned medium, cholinergic neuronal differentiation factor (CDF), causes a transition from noradrenergic to cholinergic phenotype in cultured rat sympathetic neurons. Although the transition has been known to occur in a dose-dependent manner and CDF has been purified, the examination of a complete dose-response of neurons to CDF has not been possible because sufficient quantities of pure CDF have not been available. A complete dose-response curve is essential for evaluating the biological response of the neurons, for assessing the physiological role of CDF and for understanding the mechanism of action of CDF. We report here an immunoaffinity-purification procedure for CDF with a 73.1% recovery using antibodies raised against a synthetic peptide homologous with the N-terminal region of CDF. This method produced pure CDF in quantities sufficient for examination of the full dose-response range of the neurons. Our main findings are the following. The dose-responses of acetylcholine and catecholamine metabolisms to CDF are different, although the same molecule affects both transmitters. While the half-maximal concentrations for acetylcholine induction (0.20 nM) and for catecholamine suppression (0.28 nM) are similar, the response of catecholamine metabolism begins slowly and saturates at a CDF concentration (5-20 nM) considerably higher than that of acetylcholine (0.6 nM). This may indicate that CDF affects multiple processes in catecholamine metabolism.

Amino Acid Sequence↗
PubMed1991

Follow-up of asymptomatic HTLV-I carriers among blood donors in Kyushu, Japan.

We examined mortality from adult T-cell leukemia/lymphoma (ATL/ATLL) and other diseases alleged to be associated with human T-lymphotropic virus type I (HTLV-I) among anti-HTLV-I antibody-positive blood donors in Kyushu, Japan. During 1984-87, a total of 3,991 blood donors aged 40 years or over were followed from the date of donation to the date of death or the end of the study. Crude mortality rates from ATL (with 95 percent confidence intervals) were 68 per 100,000 (13-202) for males and 36 per 100,000 (3-132) for females. The rates were underestimated by approximately 50 percent because of self-selection and short observation periods. Neither death rates from other cancers nor death rates from all cancers were elevated.

Adult↗
PubMed1989

The cholinergic neuronal differentiation factor from heart cells is identical to leukemia inhibitory factor.

A protein secreted by cultured rat heart cells can direct the choice of neurotransmitter phenotype made by cultured rat sympathetic neurons. Structural analysis and biological assays demonstrated that this protein is identical to a protein that regulates the growth and differentiation of embryonic stem cells and myeloid cells, and that stimulates bone remodeling and acute-phase protein synthesis in hepatocytes. This protein has been termed D factor, DIA, DIF, DRF, HSFIII, and LIF. Thus, this cytokine, like IL-6 and TGF beta, regulates growth and differentiation in the embryo and in the adult in many tissues, now including the nervous system.

Amino Acid Sequence↗
PubMed1989

Prevention of transmission of human T-lymphotropic virus type 1 (HTLV-1) through transfusion, by donor screening with antibody to the virus. One-year experience.

To prevent the transmission of human T-lymphotropic virus, type 1 (HTLV-1) during blood transfusion, a program was implemented to screen donors for antibodies to the virus, using a newly developed, passive agglutination (PA) method. During the period April 1986 to March 1987, 675 recipients of donor blood in whom the antibody to HTLV-1 was not present before transfusion were followed for at least 50 days after transfusion. One of these 675 seroconverted despite the transfusion of screened blood, but this seroconversion rate (0.15%) represents a marked decrease from the rate of 8.3 percent prevalent before donor screening began. The rate in the Fukuoka area of donors seropositive for anti-HTLV-1 is 5.34 percent, as detected by the PA method and 1.80 percent, as assessed by the indirect immunofluorescence (IF) technique, with PA-positive but IF-negative blood units thus accounting for 3.5 percent (5.34-1.80) of the total blood donated. The seroconversion rate among recipients transfused with blood screened by IF (at Kyushu University Hospital only) from 1981 to 1985 was 0.41 percent, which was not significantly different from the rate of 0.15 percent observed after PA screening. The discrepancy between PA and IF in the rate of seropositivity was due, in part, to the higher sensitivity of PA in detecting anti-HTLV-1. It is proposed that all donor blood in areas where HTLV-1 is endemic be screened by PA before transfusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Agglutination Tests↗
PubMed1989

The detection of an antibody against IgA allotype A2M 2 and a study of the Am genetic marker among the Han Chinese population.

The serum of a woman was found by the Ouchterlony double-diffusion and the hemagglutination inhibition (HAI) methods to have immunoglobulin A (IgA) deficiency. Further investigation using the hemagglutination (HA) test with red cells coated with IgA myeloma proteins of different specificities showed that the serum agglutinated only IgA2-, A2M-1, and A2M 2-coated cells. The patterns of the HAI test with a reference panel confirmed the presence of two specificities. One was anti-IgA2 and the other was a rare antibody against the allotype A2M 2. The anti-A2M 2 was used for population studies. Testing of the Han Chinese population, including family studies, confirms that A2M.1 and A2M.2 have an autosomal dominant mode of inheritance and are controlled by a codominant allele. The distribution of the two Am genetic markers among the Han Chinese population demonstrated A2M.1 with a gene frequency of 0.553 and A2M.2 with a gene frequency of 0.447 (chi 2 = 0.145, 0.80 greater than p greater than 0.70).

Alleles↗
PubMed1988

The dorsal occult ganglion of the wrist and ultrasonography.

In five cases of suspected occult ganglion on the back of the wrist, an ultrasonographic examination was performed. This revealed a small hypoechogenic area on the dorsal aspect between the scaphoid and the lunate or just dorsal to the lunate. Exploration of this area in three cases confirmed the presence of an occult ganglion. In the other two cases, further aspiration revealed traces of mucinous jelly at the tip of the needle.

Adult↗
PubMed1985

Purification and partial characterization of a cholinergic neuronal differentiation factor.

The choice of transmitter made by postmitotic rat sympathetic neurons in cell culture can be controlled by the environment in which they develop. One of the differentiation signals is a protein secreted by heart cells that can induce previously noradrenergic neurons to synthesize acetylcholine and form cholinergic synapses. This change in phenotype occurs without alteration in neuronal survival or growth. The differentiation factor has now been purified at least 100,000-fold, and it is homogeneous by several criteria. (i) The cholinergic activity comigrates with a single 125I-labeled protein band of 45 kDa in one-dimensional NaDodSO4/PAGE. (ii) The biological activity comigrates precisely with a series of five 125I-labeled protein spots of 45 kDa in two-dimensional gel electrophoresis. (iii) Treatment of the 45-kDa band with endo-beta-N-acetylglucosaminidase F reduces the apparent molecular size of both the labeled protein and the biological activity to a band of 22 kDa. The data suggest that the differentiation factor is a slightly basic glycoprotein with at least six glycosylation sites.

Amino Acid Sequence↗
PubMed1984

Selective IgA deficiency: analysis of Ig production in vitro.

The cellular basis of the pathogenesis of selective IgA deficiency (SIgAD) was investigated by examining surface immunoglobulin (SmIg) and in vitro pokeweed mitogen (PWM)-stimulated immunoglobulin (Ig) synthesis and by assaying in combination the counterpart lymphocytes from individuals with SIgAD and healthy donors. Peripheral blood lymphocytes (PBL) from 14 individuals with SIgAD synthesized normal amounts of IgG and IgM but did not synthesize normal amounts of IgA. Functional defects of lymphocytes for IgA synthesis were classified into four types: (i) B-lymphocyte dysfunction, (ii) increased function of suppressor T lymphocytes (Ts), (iii) decreased function of helper T lymphocytes (Th), and (iv) B-lymphocyte dysfunction and increased Ts function. The cells bearing SmIgG, SmIgM, and SmIgD were demonstrated at normal percentage ratios in all cases by immunofluorescent staining. The cells bearing SmIgA were at normal percentage ratios in the cases of T-lymphocyte dysfunction, while in the cases of B-lymphocyte defect SmIgA-bearing cells were reduced.

Adult↗