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Biomedical subjects

S Noji

Publications and source records attributed to S Noji.

At least 145 records · Page 8Linked to original sources

The expression of actin mRNA in megakaryocytes demonstrated in paraffin sections, and epoxy resin semi-thin and thin sections by in situ hybridization.

Megakaryocytes of dog bone marrow were utilized as target cells for identifying actin mRNA expressing cells on semi-thin and thin sections. After in situ hybridization with radioisotope-labeled probes was performed on paraffin sections, gelatin capsules containing freshly prepared epoxy resin were placed on the sections. The resin was solidified and detached from the slide glass, and semi-thin and thin sections were obtained. The signals showing actin mRNA expression were detected on megakaryocytes in these sections by light and electron microscopy.

Actins↗

[Surgical repair of Scimitar syndrome with azygos connection by extracardiac conduit between right pulmonary vein and left atrium].

We present a surgical case of 41-year-old woman with Scimitar syndrome. Preoperative catheterization showed azygos connection and L-R shunt ratio of 45% without intracardiac malformations. To our knowledge, this combination has not been previously reported. At operation the right single pulmonary vein was found and drained into the inferior vena cava below the diaphragm. Because of counter clockwise rotation of the heart the distance of the scimitar vein and the left atrium was too long for direct anastomosis, a polytetrafluoroethylene tube (10 mm in diameter) was utilized for an extracardiac conduit using cardiopulmonary bypass. Postoperative course was uneventful. We conclude that this technique is effective for this syndrome with a large amount of L-R shunt and a sufficient patency is expected.

Adult↗

[Clinical experience of autotransfusion of shed mediastinal blood using a new chest drainage unit].

The autotransfusion system (ATS) of shed mediastinal blood is expected as one of the techniques for non-blood open heart surgery. The ATS which has been used previously for this purpose required such items as cardiotomy reservoir, vacuum bottle, infusion pump and was complicated to manipulate. This time we have used a new chest drainage unit, Sentinel Seal Autotransfusion System (Sherwood Medical Co.), and report our clinical experience. This unit consists of two component parts. One is a blood collection bag which is later used for reinfusion of shed blood. The other part is a regular chest drainage unit with the water seal which protects pleural cavity from atmosphere. We used this system for 24 cases of open heart surgery and studied about the hematological and biochemical changes after reinfusing the shed blood postoperatively. There were no complications and side effects related to the reinfusion. The unit we have used is simple to handle and is useful when returning shed blood. We believe that this unit is quite safe for clinical use and that it will become a new strong support for non-blood open heart surgery.

Blood Chemical Analysis↗

Expression of hepatocyte growth factor gene in endothelial and Kupffer cells of damaged rat livers, as revealed by in situ hybridization.

Hepatocyte growth factor (HGF) has been demonstrated to be synthesized and secreted by non-parenchymal liver cells for liver regeneration after hepatic injury. We performed in situ hybridization to identify HGF-producing cell types in rat liver hepatitis induced by administrating carbon tetrachloride as a hepatotoxin. We found that transcripts of the HGF gene are localized in the Kupffer and endothelial cells in normal livers and increased remarkably in the Kupffer cells of the damaged livers. Thus, HGF is concluded to be synthesized in the Kupffer and endothelial cells to repair the liver tissue in paracrine fashion. No significant increase in the transcripts of the HGF gene was observed in livers after partial hepatectomy, indicating that a mechanism on liver regeneration after the hepatectomy differs from that on liver repairs. Since the HGF gene expression was also found in lung and kidney, HGF may be a ubiquitous factor for tissue repairs.

Animals↗

Expression of retinoic acid receptor genes in neural crest-derived cells during mouse facial development.

Retinoic acid (RA) is known as a teratogen that induces abnormalities in facial structures which are made up mainly of neural crest-derived mesenchyme. We investigated expression patterns of RA receptor (RAR) genes (subtypes alpha, beta, gamma) during mouse facial development. The expression of the RAR beta gene is specific for the mesenchyme around developing eyes and nose, whereas the RAR gamma gene is expressed in the mesenchyme differentiating to facial cartilages and bones. In contrast, the RAR alpha gene is expressed weakly and uniformly over the facial region. These results suggest that crucial roles of endogenous RA in facial development depend on differential functions of the RAR subtypes.

Animals↗

Expression pattern of acidic and basic fibroblast growth factor genes in adult rat eyes.

Although the retinal angiogenic and mitogenic factors have been identified to be acidic and basic fibroblast growth factors (aFGF and bFGF), little information has so far been available about the cells producing them and their function in retinal tissues. We found, by in situ hybridization, that the expression pattern of the aFGF gene differed remarkably from that of the bFGF gene in adult rat eyes. Our results demonstrated that the aFGF gene was produced by photoreceptor visual cells, neuronal cells in the inner nuclear layer and ganglion cells of the retina, in addition to pigment epithelial cells of the choroid, iris and ciliary body, and epithelial cells of the cornea, conjunctiva and lens, while bFGF was synthesized solely by the photoreceptor visual cells.

Animals↗

Functional modes of retinoic acid in mouse osteoblastic clone MC3T3-E1, proved as a target cell for retinoic acid.

Mouse osteoblastic clone MC3T3-E1 was proved as a target cell for retinoic acid (RA) in bone tissues through the demonstration of RA-receptor gene expression by the northern blot analysis. The effect of RA on the cell growth of MC3T3-E1 was repressive for both subconfluent and confluent growth, whereas RA enhancement of alkaline phosphatase expression was observed at the confluent stage. This implies that RA is a regulatory factor leading osteogenesis of the cells after the confluent stage. RA exhibited simultaneously the stage-dependent effects on EGF-dependent mitogenesis: promotive at the subconfluent, but repressive at the confluent stage.

Alkaline Phosphatase↗

No major defect detected in the gene of familial hypo-retinol-binding proteinemia.

Retinol-binding protein (RBP), a plasma protein with a molecular weight of 21,000 daltons, binds to retinol with a 1:1 molar ratio and transports it to the peripheral tissues. Familial hypo-retinol-binding proteinemia (hypo-RBPnemia) was detected in a 2-year-old girl who developed keratomalacia, and in her mother and sister. They persistently showed half the normal levels of RBP and retinol. Restriction fragments of leukocyte DNA, created by digestion with BamHI, EcoRI and PstI when human RBP complementary DNA was used as a probe, showed the same patterns in both the affected and unaffected family members. These results indicate that familial hypo-RBPnemia could be attributed to such minor changes as point mutations, rather than large deletion or insertion in the RBP gene.

Blotting, Southern↗

[A case report of surgical treatment of prosthetic valve endocarditis in tricuspid position].

A case of right sided prosthetic valve endocarditis in tricuspid position was presented. This 38-year-old male underwent tricuspid valve replacement (TVR) with SJM (31 mm) prosthetic valve because of tricuspid valve endocarditis caused by staphylococcus epidermidis five years ago. At the beginning of this year, 1989, he was admitted with complain of high fever and bloody sputa. Doppler color imaging showed prosthetic valve was stenotic whose pressure gradient was 16 mmHg in peak pressure gradient, but vegetation was not identified. Following with gastrectomy perhaps due to AGML, re-TVR was done with Carpentier-Edwards vale (31 mm). He was discharged without any problems. It seems important to determine the timing of re-operation before leading any complications.

Adult↗

Expression of retinoic acid receptor genes in keratinizing front of skin.

We found, by an in situ hybridization method with riboprobes synthesized from human cDNA of the retinoic acid receptor (RAR), that the RAR genes (predominantly gamma-subtype) are intensively expressed in the epidermis of normal and psoriasic human skins, and also in keratinizing fronts of 4-day-old mouse skins, nail matrices and hair follicles. Thus, target cells of retinoic acid in the skins are concluded to be keratinocytes, which is quite consistent with the fact that retinoic acid regulates keratinization of epidermis in vivo and also modulates expression of the keratin gene in vitro.

Animals↗

Spatial and temporal expression pattern of retinoic acid receptor genes during mouse bone development.

Spatial and temporal expression pattern of retinoic acid receptor (RAR) genes was investigated in mouse finger bones during development by an in situ hybridization method with riboprobes synthesized from a human cDNA of the RAR-alpha. We found that the RAR genes are expressed intensively and specifically in calcifying fronts of the mouse finger bones, whereas the expression pattern is rather uniform in the limb buds and cartilage matrices of the embryonic fingers. Our findings are consistent with the fact that vitamin A is essential for normal mammalian bone development.

Animals↗

The narK gene product participates in nitrate transport induced in Escherichia coli nitrate-respiring cells.

The nucleotide sequence of the Escherichia coli narK gene, which is located in the upstream region of the narCHJI operon, was determined. The narK gene encodes a very hydrophobic protein with 463 amino acid residues (Mr 49,693). A narK deletion mutant, under conditions for the induction of nitrate respiration, was unable to perform nitrate transport. Loss of transport activity was recovered by transforming the mutant with a narK+ plasmid. Thus, we conclude that the narK gene encodes a transmembrane protein participating in nitrate transport. In the narK promoter region, we defined a unique sequence that we designate as a 'nitrate box', functioning as a putative NarL-binding site, in addition to the consensus sequence of the 'anaero-box'.

Amino Acid Sequence↗

The narX and narL genes encoding the nitrate-sensing regulators of Escherichia coli are homologous to a family of prokaryotic two-component regulatory genes.

The nucleotide sequence of a 4.4-kilobase SacII-SspI fragment encoding the narXL operon and a part of the narK gene of Escherichia coli has been determined. The narX and narL genes encode proteins of molecular weight 67,275 and 23,927, respectively, and are transcribed from a common promoter, narXp, locating within 429 bases upstream of narX. Transcription from narXp is not significantly induced by nitrate under anaerobiosis, whereas transcription from narK promoter, which overlaps narXp region and is transcribed divergently, is fully induced by nitrate. The N-terminal two-thirds of the NarL protein has extensive homology with those of a diverse set of prokaryotic regulatory proteins, including OmpR, PhoB, SfrA, UhpA, CheY, CheB, NtrC, DctD, FixJ, VirG, SpoOF, and SpoOA. A segment locating in the C-terminal half of the NarL protein seems to have potential most likely to form the helix-turn-helix structure characteristic of a class of DNA-binding protein. The protein is considered to play a role as a transcriptional activator of the nitrate reductase operon, narCHJI, and the narK gene. The C-terminal region of the NarX protein also has homology with other regulatory proteins known as counterparts of two-component regulatory systems, such as EnvZ, PhoR, PhoM, CpxA, NtrB, DctB, FixL, and VirA. Presence of two copies of hydrophobic segments in the N-terminal half of the NarX protein suggests the role as a transmembrane receptor sensing nitrate.

Amino Acid Sequence↗

A monoclonal antibody against COOH-terminal peptide of human liver manganese superoxide dismutase.

Three stable hybridoma cell lines producing monoclonal antibodies specific for human liver manganese superoxide dismutase were established, and one monoclonal antibody, PG 11, was chosen for immunochemical studies. Immunoblotting demonstrated that the monoclonal antibody binds exclusively to the manganese superoxide dismutase. Immunohistochemical studies indicated that the enzyme is localized in the matrix of human liver mitochondria. To localize antibody-binding epitope, synthetic peptides of the NH2-terminal (residues 1-16) and COOH-terminal (residues 182-189, 190-196, and 182-196) parts of the enzyme were synthesized, and then their effects on the binding were studied using an enzyme-linked immunosorbent assay method. All of the above COOH-terminal peptides inhibited the binding whereas the NH2-terminal ones did not, indicating that PG 11 recognizes several peptides of COOH termini of manganese superoxide dismutase. This is the first report of monoclonal antibodies against human manganese superoxide dismutase with a distinct epitope and of the immunocytochemical demonstration of manganese superoxide dismutase.

Animals↗

Elevation in metallothionein messenger RNA in rat tissues after exposure to X-irradiation.

Metallothionein (MT) mRNA levels in tissues were measured in rats following whole-body X-irradiation (2 and 20 Gy). When compared with control rats, the elevation in MT mRNA levels of liver, kidney, and thymus was observed in irradiated rats at 9 or 72 hr after irradiation. However, the elevation in MT mRNA levels was not observed in brain, spleen, lung, heart, and testis. When compared with other tissues, testicular MT mRNA levels in control rats were extremely high, and treatment with X-irradiation produced a slight decrease of testicular MT mRNA levels. Time-course experiments indicated that hepatic and renal MT mRNA reached a maximum at 6 hr after irradiation. In low-dose (2 Gy) irradiated rat, these values were returned to control values by 4 days after irradiation. However, in high-dose (20 Gy) irradiated rat, the values were not decreased to control values. These data indicate that treatment with X-irradiation produces an elevation in MT mRNA in rat tissues.

Animals↗

Dual mechanism for stimulation of glutamate transport by potassium ions in Streptococcus mutans.

An ATP-driven primary transport system operative for L-glutamate or L-aspartate in Streptococcus mutans is, through the entire pH range from 5.5 to 8.5, specifically stimulated by extracellular potassium ions. The stimulation by potassium ions observed in the low pH range between 5.5 and 7 has been interpreted to be due to potassium ion-dependent regulation of the intracellular pH (the first mechanism). In the high pH range from 7 to 8.5, on the other hand, the present study demonstrates that potassium stimulation is essentially not associated with such intracellular pH regulation. This conclusion is based on our observation that potassium stimulation in the high pH range is insensitive to a proton conductor, carbonyl cyanide-p-trifluoromethoxy-phenyl-hydrazone. Since none of the other monovalent cations, including sodium, rubidium, ammonium, and Tris ions, could replace potassium ions in significantly stimulating glutamate transport, it is most likely that the influx of potassium ions specifically cancels the membrane potential derived by movement of glutamate with the net negative charges across a membrane and thus facilitates transport (the second mechanism). The second mechanism appears to be operative even in a low pH range, in addition to the first mechanism.

Adenosine Triphosphate↗

[Localization of epidermal growth factor and nerve growth factor mRNAs in mouse submandibular gland by a simplified in situ hybridization with riboprobes].

We simplified procedures of in situ hybridization with riboprobes on serial sections of paraffin-embedded tissues. We found that when tissues were fixed with 4% paraformaldehyde, embedded in paraffin, and cut serial sections with 5 microns thick, excellent results were obtained without proteolytic, heat, or HCl treatment. With our simplified method, both EGF and NGF mRNAs were detected intensively and specifically in granular convoluted tubule (GCT) cells of a male mouse submandibular gland, but not detected in either the acini or ducts other than the GCT. Accumulation of the silver grains was localized in the basal side of the GCT cells. No significant hybridization was observed in those cells with the sense probe and in the parotid and sublingual gland even with the anti-sense probe. Since our results were not only very consistent with other data, but also much more excellent than other results obtained by in situ hybridization on the similar tissues, our simplified method should be very useful to study gene expression on tissue sections.

Animals↗

Effect of intracellular pH and potassium ions on a primary transport system for glutamate/aspartate in Streptococcus mutans.

We have studied the mechanism of L-glutamate/L-aspartate transport in a fermentative oral bacterium of Streptococcus mutans (strain Ingbritt). The transport rate stays virtually constant throughout the pH range 5.5-8.5 and followed Michaelis-Menten type kinetics. At high pH values from 7 to 8.5, transport was essentially insensitive to N,N'-dicyclohexyl-carbodiimide (DCCD), an inhibitor of ATPase, and to carbonyl cyanide-p-trifluoromethoxyphenyl-hydrazone (FCCP), an ionophore dissipating proton motive force indicating that S. mutans transports glutamate by a primary transport system at the expense of ATP or an alternative energized metabolite. At lower external pH (7-5.5), DCCD (100 microM) or FCCP (10 microM) significantly inhibited L-glutamate transport while the intracellular ATP level was hardly affected, indicating that the activity of the primary transport system was decreased at lower intracellular pH. The glutamate transport was stimulated in the presence of potassium ion at an external pH of 6. The stimulation can be explained partly by the regulation of intracellular pH with concomitant potassium ion movement.

Aspartic Acid↗