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Biomedical subjects

K Fujikawa

Publications and source records attributed to K Fujikawa.

At least 163 records · Page 9Linked to original sources
PubMed1994

Reconstruction of the extensor apparatus of the knee with the Leeds-Keio ligament.

We describe a new method for the reconstruction of ruptures of the patellar ligament or the quadriceps tendon, using the flexible open-weave polyester Leeds-Keio prosthetic ligament. Of 25 operations performed since 1985, we were able to review 18 patients (19 knees) with a mean follow-up of 3.5 years (3 to 6.5). No patient had required immobilisation of the knee after operation and the average period to return to normal activities without a walking aid was 10.5 weeks. The average range of motion was 146 degrees, and four patients could squat fully in the Japanese style. There was an extension lag (< 10 degrees) in only four patients; eight patients had some patellar crepitus which was mildly painful in three. There were no cases of infection, persistent joint effusion or rerupture of the extensor apparatus.

Adult↗
PubMed1993

Transcription factors NF-IL6 and NF-kappa B synergistically activate transcription of the inflammatory cytokines, interleukin 6 and interleukin 8.

Single binding sites for transcription factors NF-IL6 and NF-kappa B are present in the promoter of the interleukin (IL) 6 gene. Previous studies of internally deleted promoter mutants demonstrated that these two sites are important for the transcriptional regulation of this gene. In this report, we describe the synergistic activation of the IL-6 promoter by transcription factors NF-IL6 and NF-kappa B. Cotransfection of NF-IL6 with the NF-kappa B p65 subunit resulted in strong synergistic activation of an IL-6 promoter-reporter construct. Both the NF-IL6 and NF-kappa B binding sites in the IL-6 promoter were required for synergistic activation. Similar synergistic activation was observed in the IL-8 promoter, which also contains both NF-IL6 and NF-kappa B binding sites. Furthermore, we demonstrated that NF-IL6 and the NF-kappa B p65 subunit directly associated via the basic leucine-zipper domain of NF-IL6 and the Rel homology domain of p65. Since the promoters of many other genes involved in the inflammatory and acute-phase responses also contain binding sites for NF-IL6 and NF-kappa B, the cooperation between these two factors may have an important role in these responses. We also discuss the possible interplay between various viral gene products and these two factors in the process of viral infection and constitutive cytokine production.

Base Sequence↗
PubMed1993

Isolation and characterization of the human diacylglycerol kinase gene.

The 80 kDa diacylglycerol kinase (DGK) is abundantly expressed in oligodendrocytes and lymphocytes but not to a detectable extent in other cells such as neurons and hepatocytes. As an initial attempt to delineate the mechanism of the transcriptional control of the DGK gene, we have cloned from a human genomic library a 22 kb genomic fragment. The genomic clone consists of the 5'-flanking region and 17 exons coding for approx. 53% of the total exons of human DGK, including those encoding EF-hand and zinc-finger regions. The translation initiation site is located in the second exon. S1 nuclease mapping and primer extension analysis of the human DGK mRNA identified a major transcription initiation site (position +1) at 264 bp upstream from the initiator ATG. In the 5'-flanking sequence we detected a single GC box at -35 but no canonical TATA and CAAT sequences. However, the sequence starting from the cap site (AGTTCCTGCCA) is very similar to the initiator element that specifies the transcription initiation site of some housekeeping genes. In addition, the 5'-upstream region contains several putative cis-elements. Jurkat and HepG2 cells were transfected with various 5'-deletion mutants of the upstream region fused to the structural gene of chloramphenicol acetyltransferase (CAT). The CAT assay revealed that among constructs containing up to 3.4 kb of the 5'-flanking region, a fragment of 263 bp from the transcription initiation site contains a basic promoter that is active in both types of cells. Moreover, the region between -263 and -850 contains a negative element that is active in HepG2 but not in Jurkat cells. This negative element may, at least in part, be responsible for the cell type-specific expression of the DGK gene.

Animals↗
PubMed1993

Regulation of iron metabolism in HepG2 cells: a possible role for cytokines in the hepatic deposition of iron.

In chronic inflammation it is reported that serum iron is depleted and hepatic iron is increased because of reticuloendothelial system iron blockade. However, recent studies indicate that hepatic parenchymal cells increase the uptake of transferrin-bound iron after in vivo stimulation with bacterial lipopolysaccharide, suggesting that endotoxemia itself or lipopolysaccharide-induced production of inflammation-related cytokines may also be responsible for this phenomenon. In this study the actions of inflammation-related cytokines on the synthesis of iron-binding proteins (transferrin and ferritin) and transferrin receptor and the uptake of transferrin-bound iron were investigated in a human hepatoblastoma cell line, HepG2, which is the most commonly used cell line for examining the regulation of hepatic protein synthesis by cytokines. The cells were exposed to interleukin-1 beta, interleukin-6 or tumor necrosis factor-alpha separately for 24 hr. In each cytokine treatment group, the level of transferrin, which is secreted into the conditioned medium, was found to be decreased compared with that of untreated cells. On the other hand, the biosynthesis of ferritin was markedly elevated after the same treatment. This increase in ferritin by cytokine treatment was diminished when deferoxamine was used concomitantly to deplete intracellular chelatable iron. After stimulation with interleukin-1 beta, interleukin-6 or tumor necrosis factor-alpha, 59Fe-labeled transferrin uptake into the cells was increased by 36%, 48%, or 18%, respectively, and this uptake was inhibited by the addition of excess unlabeled transferrin. A binding study with 125I-labeled diferric transferrin revealed that the three cytokines increased the number of transferrin receptors on the cell surface by 1.15-fold to 1.35-fold.(ABSTRACT TRUNCATED AT 250 WORDS)

Carcinoma, Hepatocellular↗
PubMed1993

Genotoxic potency in Drosophila melanogaster of selected aromatic amines and polycyclic aromatic hydrocarbons as assayed in the DNA repair test.

Drosophila melanogaster stock consisting of meiotic recombination deficient (Rec-) double mutant mei-9a mei-41D5 males and Rec+ females was exposed at the larval stage to an aromatic amine or a polycyclic aromatic hydrocarbon. After emergence as adult flies, the males and the females were scored separately. When the treatment caused a dose-dependent reduction in the male to female ratio from the control level; the experiment was repeated with a larval stock consisting of Rec+ males and Rec+ females under comparable conditions. A preferential killing effect upon Rec- larvae was taken as evidence of DNA damaging effect of the test compound. Among 16 compounds tested, 1-AP, B(a)P, 2-AF, DAF, 4-AAF, 2-AAF, 1-AA, 2-AA, DMA, B(a)A and DMBA were registered as positive; Py and 3-MC were weakly positive; and B(e)P, Fluo and Ant were negative. The selective killing effects of the compounds in each of the pyrene, fluorene and anthracene series varied drastically as a function of structure in a way similar to that reported for the genotoxicity in Drosophila and the carcinogenicity in rodents. The Drosophila DNA repair assay will serve as a simple adjunct to the already available means for studying the genotoxic potency of aromatic amines and polycyclic aromatic hydrocarbons.

2-Acetylaminofluorene↗
PubMed1993

Abnormal hepatic iron accumulation in LEC rats.

The LEC (Long-Evans cinnamon) rat is a mutant strain displaying hereditary hepatitis and spontaneous hepatocellular carcinoma, and shows abnormal hepatic copper accumulation similar to that occurring in Wilson's disease. We evaluated the iron metabolism of LEC rats compared to LEA (Long-Evans agouti) rats. Hepatic iron and ferritin concentrations were remarkably increased depending on age in LEC rats but not in LEA rats. Increased hepatic iron is normally associated with decreased serum transferrin and total iron binding capacity in hepatic iron overload. In LEC rats, however, both serum transferrin and total iron binding capacity increased with increasing hepatic iron. This increase of serum transferrin and hepatic iron may be an additional important factor contributing to liver injury in LEC rats.

Age Factors↗
PubMed1993

Gingival epithelial cells secrete a substance which increases the collagenolytic enzyme activity of periodontal ligament cells.

To examine whether the gingival epithelium provokes a loss of connective tissue attachment during periodontitis, periodontal ligament (PDL) cells and gingival fibroblasts (GF) were cultured with conditioned medium of gingival epithelial cells, and the collagenolytic activity of PDL cells and GF were examined, respectively. The epithelial cells were cultured in the presence or absence of lipopolysaccharide (LPS), and the respective conditioned media were added to the cultures of PDL cells and GF. The collagenolytic response of PDL cells to both of the conditioned media was 3- to 10-fold higher than that of each control, whereas the response of GF was only 1.4-fold higher. The LPS-stimulated epithelial conditioned medium showed a stimulation rate for collagenolytic activity similar to that of the LPS-free conditioned medium. These results suggest that human gingival epithelial cells, without LPS stimulation, secrete a substance which accelerates the collagenolytic enzyme activity of PDL cells for degradation of PDL fibers, in the absence of GF enzyme activity.

Cell Division↗
PubMed1993

[Cytotoxic effects of diclofenac sodium].

The authors studied the cytotoxic effect of diclofenac sodium, a prostaglandin synthetic inhibitor, on Chang's cultured human conjunctival cells. Diclofenac sodium inhibited cell growth dose-dependently. Although cell growth was interrupted 12 hrs later by one minute of exposure to a 0.1% solution of diclofenac sodium, the cells began to grow again 24 hrs later. Twenty-four hours later, a one-minute exposure to a 0.1% solution of diclofenac sodium revealed no cytotoxic effects electron microscopically. The effect on the cell cycle of exposure to 0.1% diclofenac sodium was studied using a flow cytometer. Twelve hours after exposure to diclofenac sodium, DNA histograms showed a broader G1 peak, and increase in mitotic phase cells and dead cells with a low DNA content on the left of the G1 peak. 24 hrs later, the number of dead cells and DNA synthetic phase cells increased and mitotic cells gradually decreased, almost disappearing within 48 hrs.

Cells, Cultured↗
PubMed1992

[Dynamic motion study of the cervical spine using ultra-fast gradient echo with RF-spoiled GRASS--evaluation of cervical instability].

In order to assess the instability of the cervical spine, ten patients with cervical spondylosis were studied by dynamic MRI using Ultra-Fast RF-Spoiled GRASS which is capable of subsecond imaging. Dynamic MRI has proved to be useful in the evaluation of cervical instability, especially in detecting the transient abnormal mobility of cervical spines which cannot be identified on conventional radiography.

Humans↗
PubMed1992

Arteriovenous malformation of the pancreas associated with hepatocellular carcinoma. A case report and review of the literature.

A case of pancreatic arteriovenous malformation (AVM) with hepatocellular carcinoma is reported. The patient, a 56-year-old Japanese man, was asymptomatic. The pancreatic lesion was found incidentally during an evaluation for hepatocellular carcinoma. Celiac arteriogram demonstrated tortuous feeding arteries, a racemose intrapancreatic stain, which disappeared before the venous phase, and early portal filling.

Arteriovenous Malformations↗
PubMed1992

[Determination of human erythrocyte membrane Na+/K(+)-ATPase activity in small volume of blood sample].

Na+/K(+)-ATPase of the cell membrane is considered to be closely related to the pathology of various diseases including hypertension and heart failure. The activity of this enzyme in the erythrocyte membrane has been determined in earlier reports by the assay of inorganic phosphate generated from the substrate ATP or radioimmunoassay after binding 3H ouabain to the erythrocyte membrane, using a large volume of blood samples. However, as neither method was appropriate for wide routine use, we developed a method to assay this enzyme in a small volume (10 ml) of fresh human blood samples with re-evaluation of conditions for the inorganic phosphate assay. In this method, the coefficient value (CV) of membrane protein amount and the NA+/K(+)-ATPase activity were 2.2% and 2.5% respectively, indicating sufficient precision of the assay. Moreover, in 97 subjects without abnormalities in blood biochemical tests (77 males and 20 females) aged 35-59 years, the enzyme activity showed no differences according to age or sex, ranging from 0.217 to 0.071 mumols Pi/mg/hr with a mean of 0.130.

Adult↗
PubMed1992

Circulating transferrin receptor in acute leukemias.

Serum transferrin receptor (s-TR) levels in acute leukemia patients were measured by a recently developed sandwich radioimmunoassay. The mean s-TR level for normal subjects (n = 205) was 246 +/- 79 (mean +/- 1 SD) ng/ml. The values for patients with untreated acute myelocytic leukemia (AML, n = 18) and untreated acute lymphocytic leukemia (ALL, n = 14) were 398 +/- 175 ng/ml and 479 +/- 176 ng/ml, respectively, both of which were significantly higher than those for the normal subjects (AML, p < 0.02; ALL, p < 0.05). When complete remission was achieved with initial remission induction therapy, s-TR decreased to 262 +/- 47 ng/ml (n = 22, 12 AML and 10 ALL), returning to normal levels. There was a good correlation between s-TR levels and the number of leukemic cells in peripheral blood (r = 0.743, n = 32, p < 0.01). In two patients with AML, serial changes of s-TR values and numbers of blast cells in the peripheral blood and bone marrow occurred in a parallel manner. If, therefore, this assay for s-TR can be made more sensitive, it may become useful for assessing acute leukemia activity and for monitoring the effects of therapy.

Acute Disease↗
PubMed1992

[Synergism between transcription factors NF-IL6 and NF-kappa B in IL-6 gene regulation].

NF-IL6 and NF-kappa B are nuclear proteins supposed to play an important role in the regulation of acute-phase protein synthesis and inflammatory response against infection and tissue injury as a host defence mechanism. In addition the promoter region of the interleukin-6 (IL-6) gene has a NF-kappa B binding motif as well as a NF-IL6 binding site. Considering of these facts, we come to investigate that there may be a synergistic effect between NF-IL6 and NF-kappa B in the regulation of IL-6 gene expression. In order to study it, some combinations of expression vectors NF-IL6 cDNA, NF-kappa B (p50/p65) cDNA and reporter plasmid K18-CAT which contains human IL-6 promoter linked to the chloramphenicol acetyltransferase (CAT) gene, were transfected into Jurkat cells and the CAT activities were examined. Co-transfection of NF-IL6 and NF-kappa B (p50/p65) cDNA revealed a dramatic increase of acetylated [14C] chloramphenicol, and its CAT activity reached to 40%. Then, co-transfection of NF-IL6 and NF-kappa B subunit p65 alone showed a high level of CAT activity, too. When 5' deletion mutant reporter plasmid K9-CAT lacking the NF-IL6 binding site was used, co-transfection of NF-IL6 and NF-kappa B (p50/p65) showed low level of CAT activity. These results indicate that there is a synergistic effect between NF-IL6 and NF-kappa B (p50/p65) in IL-6 gene regulation. Among two subunits of NF-kappa B (p50/p65), p65 seems to play an important role rather than p50 does in synergism between NF-IL6 and NF-kappa B. Besides, this synergistic function comes to work only when NF-IL6 binds to its binding site of IL-6 promoter region.

Binding Sites↗
PubMed1992

[An experience with augmentation sigmoid cystoplasty for urinary incontinence caused by sacral agenesis: a case report].

Sacral agenesis is an uncommon disease. About 50 cases have been reported in Japan since 1929. Neurogenic bladder is often accompanied with the disease. The patient was a 26-year-old man who had suffered from persistent urinary incontinence since his childhood. Kidney-ureter-bladder (KUB) revealed Type IV sacral agenesis according to the classification by Renshaw. The upper urinary tract remained normal. Urodynamics study showed a low compliance bladder with low urethral pressure. Pharmacotherapy failed to improve his continence. Augmentation sigmoid-cystoplasty was undertaken to enlarge vesical capacity and it has successfully overcome his urinary incontinence. Clinical aspects of sacral agenesis are discussed focusing on urological problems.

Adult↗
PubMed1992

[A flow cytometric study of the effects of benzalkonium chloride on the cell cycle].

The effects of benzalkonium chloride (BAK) on the cell cycle of Chang's cultured human conjunctival cells were investigated by a flow cytometer (FCM). The cells were exposed to BAK solutions for 60 sec and after 12-48 hrs were fixed in 20% ethanol, treated by 0.25% RNase and stained by 0.005% propidium iodide. DNA histograms were analyzed by the FCM. As a result, although many cells were damaged by exposure to solutions of 0.0025% or 0.005% BAK, they began to grow again 48 hrs later. BAK decreased red fluorescence intensity in DNA histograms. The histogram shifted to the left 12 hrs after the cells were exposed to 0.0025% or 0.005% BAK solutions and recovered 48 hrs later. The DNA synthetic phase of the cell cycle was inhibited by exposure to solutions of 0.0025% BAK and then recovered 48 hrs later.

Benzalkonium Compounds↗