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Biomedical subjects

K Yamanouchi

Publications and source records attributed to K Yamanouchi.

At least 73 records · Page 4Linked to original sources

[Pulmonary aspergilloma clearly demonstrated by digital subtraction bronchography].

A 69-year-old woman was admitted because of persistent productive cough, low-grade fever and abnormal pulmonary infiltrates. Chest roentgenograms revealed right pulmonary cavitary lesions and infiltrates in both upper pulmonary fields. Digital subtraction bronchography (DSBG) was performed to detect for the presence of pulmonary cavitary lesions. Double-contrast DSBG images clearly revealed marked bronchiectasis in the right pulmonary cavity with aspergilloma in the cavitary lesions. Aspergilloma was not obvious in routine chest roentgenograms and computed tomograms. Asperillus fumigatus was cultured from sputum materials, and aspergilloma of the pulmonary cavity was clinically diagnosed. The infiltrative lesions were thought to be associated with invasive aspergillosis because the patient exhibited persistent low-grade fever and positive inflammatory reactions. Anti-fungal chemotherapy was highly effective in treating these lesions. DSBG is a new, less-invasive bronchographic technique for the evaluation of bronchial trees, and in this case proved useful for the detection of small aspergilloma in pulmonary cavities.

Aged↗

Equine inhibin/activin beta A-subunit mRNA is expressed in the endometrial gland, but not in the trophoblast, during pregnancy.

The expression of both inhibin alpha- and inhibin/activin beta A-subunit mRNA was examined in equine uteroplacental tissues collected during pregnancy (days 90 to 300). Northern blot analysis revealed that 5 transcripts (7.0, 4.1, 3.4, 2.6, 1.5 kb) of beta A-subunit were present, and the most abundantly expressed transcript was the 1.5 kb one. Relatively high levels of the 1.5 kb transcript were seen in the second trimester of pregnancy compared to what was found in the third trimester. To identify the tissue localization of beta A-subunit mRNA, in situ hybridization was performed, and the positive signal was observed exclusively in the endometrial glands, but not in the fetal placental tissue (trophoblast) at days 150, 210, and 300 of pregnancy. On the other hand, inhibin alpha-subunit transcript could not be detected at any stage of pregnancy examined either by Northern blot analysis or in situ hybridization. Although the factor(s) regulating the gene expression of beta A-subunit in this equine tissue is currently unknown, these results suggest that activin, but not inhibin, is predominantly produced in the endometrial glands of the pregnant mare, and thus produced activin may play a paracrine or endocrine role during pregnancy in this species.

Animals↗

Lordosis in male rats: effect of dorsal raphe nucleus cuts.

The efferents and/or afferents of the dorsal raphe nucleus (DRN) were transected by several types of cut in castrated male rats, and lordosis behavior was observed after implantation with Silastic tubes containing estradiol. Throughout the behavioral tests, low incidences of lordosis were observed in control male rats without brain surgery or with a sham operation. In contrast, all male rats with a horizontal circle cut at the ventral area of the DRN displayed lordosis, and the mean lordosis quotient (LQ) was higher than that in control rats, while rats with a horizontal cut at the dorsal area of the DRN did not. Furthermore, mean LQs in male rats with an anterior half-circle horizontal cut at the ventral area of the DRN were higher than those in control groups. A posterior half-circle cut at the ventral area had no effect. In addition, male rats with a half-dome cut located anterior to the DRN showed a high LQ score, but rats with a posterior half-dome cut did not. These results suggest that anterior and anteroventral neural fibers of the DRN are involved in the lordosis inhibiting mechanism in male rats.

Afferent Pathways↗

Histopathological features of canine distemper recently observed in Japan.

Eleven dogs with canine distemper (CD) from the Chubu region of Japan and the Tokyo area were examined. Clinically, respiratory and neurological signs were present in all animals. Histopathologically, all showed characteristic CD lesions of bronchopneumonia and demyelinating encephalitis. However, some differences in gastrointestinal abnormalities were observed. Three out of four dogs from the Chubu region had severe diarrhoea and gastroenteritis, associated with numerous eosinophilic inclusion bodies in the mucosal epithelia. The remaining dog from this area showed vomiting, but not diarrhoea, and also had a number of intraepithelial inclusion bodies in the gastric and intestinal mucosa. In contrast, the seven dogs from the Tokyo area showed neither gastrointestinal symptoms nor intraepithelial inclusions in the stomach or intestine. Immunohistochemical examination for CD virus antigen, however, revealed that these seven dogs had immunoreactive products in the mucosal epithelia, suggesting that the epithelial cells had either a low level of infection with CD virus or were infected with a less cytopathogenic virus. These findings suggest that the dogs in this study were probably affected by two distinct types of CD, in terms of epitheliotropism and cytopathogenic effects on the gastrointestinal tissues.

Animals↗

Effects of p-chlorophenylalanine on reflexive and noncontact penile erections in male rats.

To clarify the role of serotonin in penile erection, testosterone-primed castrated male rats were treated with the serotonin-synthesis inhibitor, p-chlorophenylalanine (pCPA), and reflexive erection (RE; male supine, penile sheath retracted) and noncontact erection (NCE; penile erection evoked by remote sexual stimuli) tests were performed. Half the males were injected with 100 mg/kg pCPA 4 times before each test; control males were treated with saline instead of pCPA. In the RE test, compared to the control group, pCPA-treated males had a shorter erection latency, but they also displayed fewer erections. NCE tests were conducted as a 2 x 2 factorial experiment: pCPA or saline, and estrous female present or absent. Only the pCPA-female Group had a high proportion of responders (68%), compared to 14-27% in the other Groups (p < 0.02). These results suggest that the serotonergic system exerts facilitative and inhibitory influences on different systems in regulating reflexive erection. On the other hand, serotonin appears to play an inhibitory role in the induction of noncontact erection, because pCPA did not directly induce erection, but rather facilitated the response to females.

Animals↗

Efficient selection of preimplantation transgenic embryos by an improved procedure using Dpn I-Bal 31 digestion and the polymerase chain reaction.

Efficient selection of preimplantation transgenic embryos by an improved method after pronuclear injection of exogenous DNA is described. The method is based on subjecting DNA extracted from the embryos to restriction enzymes as well as the polymerase chain reaction (PCR). The incorporated procedure included recovery of the digested DNA with glassmilk before PCR, which markedly enhanced the rate of accurate detection of transgenic embryos. When exogenous DNA sequences in the mouse embryos were not integrated into the genome they were digested with both Dpn I and Bal 31, and subsequent PCR analysis generated DNA fragments of the injected DNA sequence in only 1.5% of cases examined. However, DNA extracted from mouse embryos containing the transgene sequences integrated into the genome evaded digestion by both enzymes and yielded transgene-specific PCR products in 68.6% of the embryos tested. When bovine embryos were used, sequences of the endogenous haemoglobin gene used as a control genomic DNA sequence were protected from enzyme digestion (PCR products in 70.5% of the embryos examined); by contrast, the non-integrated injected sequences were almost completely eliminated by the same treatment (PCR products in 1.4% of the embryos examined). It is suggested that this method might be useful for the selection of transgenic embryos before embryo transfer, thereby reducing the number of recipient females required.

Animals↗

Different female reproductive phenotypes determined by human growth hormone (hGH) levels in hGH-transgenic rats.

The effect of continuous human GH (hGH) secretion on female reproduction was studied in adult female transgenic rats expressing the hGH gene with a mouse whey acidic protein (mWAP) promoter. Two lines of transgenic female rats carrying the mWAP/hGH gene were established and used in the study. One was characterized by relatively high levels of serum hGH (high line), and the other had relatively low levels (low line). High-line female rats had recurring, pseudopregnancy-like estrous cycles accompanied by increased serum progesterone levels for 2 wk after ovulation, and they were fertile. In these rats, luteinization occurred spontaneously without cervical stimulation, probably due to high levels of serum hGH, which has prolactin (PRL)-like activity in the rat. Although low-line female rats had recurring, regular 4-day estrous cycles, they were sterile. In these rats, pseudopregnancy could not be induced by mating or by mechanical cervical stimulation. PRL surges following cervical stimulation were not detected, and PRL secretion was not induced by a dopamine antagonist, metoclopramide. The ovarian tissue in this line had a much higher number of corpora lutea and grew much heavier than in normal littermates, suggesting impairment of PRL-induced structural luteolysis. Suppression of PRL secretion in low-line rats was, at least in part, due to a marked decrease in the number of lactotrophs in the pituitary. The present study shows that the serum hGH level plays a crucial role in regulating luteal function in female transgenic rats expressing the hGH gene.

Animals↗

The association of platelet-derived growth factor (PDGF) receptor tyrosine phosphorylation to mitogenic response of human osteoblastic cells in vitro.

OBJECTIVES: The purpose of this study was to make clear the relationship of human osteoblastic cell growth, induced by platelet-derived growth factor (PDGF), to PDGF receptor tyrosine phosphorylation. MATERIALS AND METHODS: Osteoblastic cells derived from human maxilla were cultured with human PDGF. The cell growth was evaluated by cell number and DNA synthesis. PDGF receptor tyrosine phosphorylation was detected by immunoblot analysis using anti-PDGF receptor alpha, beta subunits and anti-phosphotyrosine antibodies. Genistein, a tyrosine kinase inhibitor, was added to the culture to investigate the effect on osteoblastic cell growth and PDGF receptor tyrosine phosphorylation induced by PDGF. RESULTS AND CONCLUSIONS: PDGF stimulated the proliferation of human osteoblastic cells and this effect was synergetic with serum stimulation. DNA synthesis of osteoblastic cells was elevated by PDGF in a dose dependent manner at the minimum concentration of 1 ng ml-1. PDGF also induced PDGF receptor tyrosine phosphorylation within 1 min on osteoblastic cells, and tyrosine phosphorylation occurred on PDGF receptor subunits alpha and beta. Genistein inhibited cell growth and receptor tyrosine phosphorylation, which was induced by PDGF on these cells. In conclusion, human osteoblastic cell growth induced by PDGF is shown to relate to tyrosine kinase of PDGF receptors.

Becaplermin↗

Expression and cellular localization of inhibin alpha-subunit mRNA in equine fetal gonads.

The expression of inhibin alpha-subunit mRNA in equine fetal gonads during pregnancy (Days 90 to 300) was examined by means of Northern blot analysis. In all samples examined, a single species of transcript was detected at the size of 1.5 kb. A digoxigenin-labeled antisense cRNA probe specific to equine inhibin alpha-subunit was synthesized and in situ hybridization analysis to locate the inhibin alpha-subunit mRNA positive cells was performed using frozen tissue sections of equine fetal ovary (day 150 of pregnancy) and equine fetal testis (day 180 of pregnancy). In the fetal ovary, positive cells were seen throughout the interstitial area but did not show any particular localization. In the fetal testis, on the other hand, the antisense cRNA hybridized almost exclusively to the interstitial cells surrounding developing seminiferous cords and Sertoli cells within the cords. Positive signals were also detected in a limited number of the interstitial cells located away from the cords. These results suggest that in equine fetal gonads, inhibin and/or inhibin alpha-subunit related molecules such as the monomeric form are produced and these molecules may have a paracrine/autocrine role within the gonads.

Animals↗

Effects of p-chlorophenylalanine on male sexual behavior in female rats with mesencephalic raphe nuclei lesions.

The role of serotonergic neurons in the mesencephalic raphe nuclei in regulating male sexual behavior in female rats was examined. The median or dorsal raphe nucleus lesions (MRL or DRL) were made in ovariectomized rats and behavioral tests were performed after implantation of Silastic tubes containing testosterone and treatment with serotonin synthesis inhibitor p-chlorophenylalanine (pCPA). Half of the animals in each group received 4 100 mg/kg pCPA injections before the behavioral test. As a result, the incidences and frequencies of mounts and intromissive patterns in the MRL and DRL groups were comparable to those in control females without brain surgery. Mount latency in the MRL females was shorter than that in the control females. When pCPA was given, most females with or without brain surgery showed mounts and intromissive patterns, and frequencies were higher than those in females without pCPA. These results suggest that the median raphe nucleus plays an inhibitory role in the onset mechanism for mounting. On the other hand, a stronger inhibitory influence in regulating male sexual behavior exists in other serotonergic neurons than those in the median and dorsal raphe nuclei in female rats.

Animals↗

Facilitatory effect of ventral cut of dorsal raphe nucleus on lordosis in female rats.

The role of the ventral projections of the dorsal raphe nucleus (DRN) in regulating lordosis was examined in female rats. The ventral neural fibers of the DRN were transected by the horizontal circle-cut (VC), anterior (AVC) or posterior half-circle cut (PVC) at the same time as ovariectomy. Three weeks after the surgery, all the rats were implanted with a Silastic tube containing estradiol (E2). The behavioral test was started one day after E2-implantation, and 5 tests in total were carried out every other day. The results showed that the VC females had a higher lordosis quotient (LQ) than control rats. Furthermore, as well as females with the VC, a high score LQ was observed in the AVC rats but not in the PVC females, compared to those in the control animals. These results suggest that the anteroventral neural fibers of the DRN are involved in the lordosis inhibiting mechanisms in female rats.

Animals↗

Potentiation of ejaculatory activity by median raphe nucleus lesions in male rats: effect of p-chlorophenylalanine.

The effect of radiofrequency lesions in the median or dorsal raphe nucleus (MRL or DRL) on copulatory behavior was examined in sexually inexperienced male rats. Three weeks after castration and the brain surgery, all males were subcutaneously implanted with Silastic capsules containing testosterone. In the first behavioral test, the frequency of ejaculation in the MRL group was significantly higher than that in sham and DRL males, but mount and intromission were not. Seven days after the first test, the second test was carried out after treatments with 100 mg/kg p-chlorophenylalanine (PCPA), a serotonin synthesis inhibitor, or saline daily for 4 days in MRL and DRL males. The frequencies of male sexual behavior in PCPA treated DRL males were higher than those in saline treated DRL males. In contrast, even after treatments with PCPA, male sexual activity in MRL males was comparable to those in saline treated MRL males. These results suggest that serotonergic neurons in the median raphe nucleus play an inhibitory role in the regulation of male sexual activity, especially ejaculation. Furthermore, it can be thought that PCPA acts on the median raphe neurons and facilitates ejaculatory behavior.

Analysis of Variance↗

Cell-cell contact down-regulates expression of membrane type metalloproteinase-1 (MT1-MMP) in a mouse mammary gland epithelial cell line.

Membrane type matrix metalloproteinase (MT-MMP), which possesses a C-terminal transmembrane domain, is expressed on the cell membrane (Sato et al., 1994, Nature 370: 61-65). It was suspected, therefore, that the expression of MT-MMPs might be regulated by cell-cell interactions. We examined the patterns of MT1-MMP expression in a mouse mammary gland epithelial cell line, HC11, which is capable of responding to prolactin in vitro. HC11 cells form well-differentiated monolayer of cuboidal epithelium at confluence. During the log growth phase, cells which are well dispersed and seemingly migrating actively, or located at the periphery of small colonies, reacted strongly with an anti-MT1-MMP antibody, whereas no MT1-MMP immunoreactivity was detected in the cells which established cell-cell contact with adjacent cells. At confluence, the HC11 cells lost MT1-MMP immunoreactivity completely. Northern blot analysis revealed that MT1-MMP mRNA is present at a high level in HC11 cells during the log phase of growth. Although MT1-MMP immunoreactivity disappeared by the 1st day confluence was reached, the decline of MT1-MMP mRNA levels started only a few days later. The discrepancy in the timing of decrease of MT1-MMP protein and that of the transcripts suggests the presence of translational control mechanisms for MT1-MMP expression during cell-cell interaction.

Animals↗

[Slow virus infection].

Subacute sclerosing panencephalitis (SSPE) and progressive multifocal leukoencephalopathy (PML) are the major slow virus infections of humans. SSPE is caused by mutant measles virus after long persistence in the brain. Its pathogenesis is discussed, based on accumulating data on molecular properties on measles virus prevalent in the field as well as recent identification of measles virus receptor. Current situation on the therapy of SSPE is also described. PML is caused by the infection of oligodendroglia with persisting JC virus. Its pathogenesis is discussed based on the molecular properties of the archetype and PML type of JC virus, and on the recent findings on the involvement of JC virus T antigen in demyelination. Increase in the incidence of PML in patients with AIDS is discussed in view of the interactions between HIV and JC virus.

Humans↗

Secretory production of recombinant urokinase-type plasminogen activator-annexin V chimeras in Pichia pastoris.

To produce a thrombi-targeting plasminogen activator, we expressed a fused gene that contains a modified pre-sequence of Mucor pussilus rennin (MPR) followed by a chimeric gene of single-chain urokinase-type plasminogen activator (scu-PA)::annexin V (AV). The fused gene was ligated into an integrative vector, under the control of the alcohol oxidase 1 (AOX1) promoter (p), and transformed into Pichia pastoris. Transformants were monitored for the secretion of fibrinolytic activity. The highest expressing clone, HB225, secreted as much as 600 international units (IU) of fibrinolytic activity per ml of culture medium under optimal conditions. It contained three tandem copies of the full-size vector disruptively integrated into the AOX1 sequence. Western blot analysis revealed that the secreted chimera was highly susceptible to proteolysis. Addition of excess amino acids (aa) to the culture medium minimized the degree of proteolysis. Two major species of chimera, 85 and 65 kDa, were then isolated from the culture medium. The former was the intact form consisting of a single-chain and showing full enzyme activity after activation by plasmin. The latter was an enzymatically processed form consisting of two chains held by a disulfide bond, having full enzyme activity without activation. Both chimeras exhibited calcium-dependent phospholipid (PL)-binding affinities similar to the parent AV.

Amino Acid Sequence↗

Nucleotide sequences of the M gene of prevailing wild measles viruses and a comparison with subacute sclerosing panencephalitis virus.

We determined the nucleotide sequences of the coding region for the M gene in seven strains of measles virus (MV) that were isolated in Japan between 1984 and 1993. The mutation found among the seven differed from those of laboratory strains. Many of these mutations were the same as those that are characteristic of SSPE viruses. Thus, we suggest that the mutations that have been considered specific to SSPE virus are in fact consensus among prevailing MV.

Base Sequence↗

Lordosis-inhibiting effect of progesterone in female rats with lesions in septum, preoptic area, or dorsal raphe nucleus.

Radiofrequency lesions in the septum (SL), the preoptic area (POAL), or the dorsal raphe nucleus (DRL) were made in ovariectomized rats. In a control group of 16 females, ovariectomy, but no brain surgery, was performed. All animals except half of the control rats received injections of 5 mg progesterone (P) 1 h prior to the injection of 5 micrograms/kg b.w. of estradiol benzoate (EB). Instead of 5 mg P, oil was administered to half of the controls. Forty-four hours after EB, all females received 0.5 mg P. A sexual behavior test was performed 4 h after the last injection of P. The result was that oil-treated control rats showed high lordosis quotient (LQ) and soliciting behavior. In contrast, low scores of LQ and no soliciting behavior were observed in all of the 5 mg P-treated rats, even if the SL, POAL, or DRL was made. These results suggest that the septum, the preoptic area, and the dorsal raphe nucleus are not essential for the female sexual behavior-inhibiting mechanisms of progesterone.

Animals↗