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Biomedical subjects

T Okuno

Publications and source records attributed to T Okuno.

508 records · Page 29Linked to original sources

Identification and characterization of human herpesvirus 8 open reading frame K9 viral interferon regulatory factor by a monoclonal antibody.

OBJECTIVE: To identify and characterize an open reading frame (ORF) in association with its mRNA and a coding protein recognized by anti-human herpesvirus 8 (HHV-8) monoclonal antibody B291. STUDY DESIGN/METHODS: The antigen detected by B291 was characterized by immunofluorescence method, Western blot analysis, laser confocal microscopy, and immunohistology cDNA of the protein was identified by immunoscreening the HHV-8 cDNA library with B291 and sequenced. Structure and kinetics of the mRNA expression was investigated by 5',3' rapid amplification of cDNA ends (RACE) and Northern blot analysis. RESULTS: Viral interferon regulatory factor (vIRF; approximately 50 kd) was a protein recognized by B291. It was detected in 12-O-tetradecanoyl phorbol-13 acetate (TPA)-induced, but not uninduced, HHV-8-infected cell lines (BCBL-1 and BCP-1) and the tumor cells obtained from a SCID mouse injected with uninduced BCBL-1. The expression of vIRF was predominantly in the nucleus and was partially diminished by phosphonoformic acid (PFA) treatment, whereas the mRNA (approximately 2.4 kb) was accumulated. CONCLUSIONS: vIRF is an early protein expressed in the nucleus and may be involved in Kaposi's sarcoma tumor formation.

Amino Acid Sequence↗

Measurement of protein C inhibitor in seminal plasma is useful for detecting agenesis of seminal vesicles or the vas deferens.

Protein C inhibitor (PCI), a plasma serine protease inhibitor of activated protein C, is present at high concentrations in the seminal plasma of normal subjects and is decreased in some infertile patients. We measured the concentrations of PCI, prostate-specific antigen, and fructose in the seminal plasma of infertile patients (n = 125) and of normal subjects (n = 13). We also measured time-dependent changes in the concentrations of PCI and fructose in seminal plasma after ejaculation. A weak correlation was found between the levels of PCI and fructose (r = 0.268, P = 0.016). The PCI level in seminal plasma of patients with seminal vesicle and/or vasal agenesis was significantly lower (P < .01) than in normal subjects. The level of fructose in seminal plasma decreased in vitro in a time-dependent manner after ejaculation, whereas the concentration of PCI was stable at 48 hours after ejaculation. These data suggest that PCI in seminal plasma, as well as fructose, may become one of the markers for agenesis of seminal vesicles and/or the vas deferens.

Enzyme-Linked Immunosorbent Assay↗

Functional analysis of Kaposi's sarcoma-associated herpesvirus RTA in an RTA-depressed cell line.

OBJECTIVE: The viral transcriptional activator encoded by Kaposi's sarcoma-associated herpesvirus (KSHV) open reading frame 50 (RTA) is expressed in the immediate-early phase of reactivation. We isolated an RTA-depressed cell line, BLS50-4, by subcloning from the KSHV-infected cell line, BCBL-1. RESULTS: In addition to RTA, induction of some lytic gene expressions was also remarkably reduced in BLS50-4 cells, but that of the K8.1 and ORF65 gene expressions was not. Both the replication of the KSHV genome and the release of KSHV DNA into the medium were greatly reduced. Transfection of RTA into BLS50-4 cells restored the expression of K9 (vIRF) and ORF59, but not K8.1. CONCLUSIONS: Thus, we showed that expression of late genes was not directly controlled by RTA, unlike the other groups concluded. We also showed that, by isolating the RTA-depressed cell line, the RTA protein had a critical role in viral DNA replication and the expression of several lytic genes.

Cell Line↗

Endolymphatic hydrops in perilymphatic fistula.

Cochlear hydrops was produced either by injecting artificial perilymph into the subarachnoid space or by sucking perilymph through the round window membrane. The animals were either vitally fixed immediately or kept alive for 1 to 3 months before fixation. Conventional celloidin embedding method was used for serial sectioning. Animals with cochlear hydrops were selected from among 56 guinea pigs; 13 animals were used for the present observation. The cochleas of these animals showed slight to moderate hydrops, while the vestibule and semicircular canals demonstrated either a normal or collapsed endolymphatic space. The pars superior does not develop hydrops. Changes in the saccular wall were not pronounced. One animal developed marked cochlear hydrops with atrophy of both Corti's organ and stria vascularis. The vestibule and semicircular canals showed marked collapse. The ductus reuniens and the saccule were completely obstructed. Marked cochlear hydrops was produced by blocking the longitudinal flow of endolymph. It is assumed that there may be cochlear hydrops in clinical cases of perilymphatic fistula, with various pathologies of the endolymphatic space of the vestibule and semicircular canals.

Animals↗