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Biomedical subjects

K Yamanouchi

Publications and source records attributed to K Yamanouchi.

At least 55 records · Page 3Linked to original sources

Expression of milk protein genes is induced directly by exogenous STAT5 without prolactin-mediated signal transduction in transgenic mice.

STAT5 is a member of the family of STAT (signal transducer for activating transcription), which was isolated from nuclear extracts of the lactating gland of sheep. It was reported that, in vitro, STAT5 could be activated directly by prolactin (PRL) without PRL-mediated signal transduction. The present study was conducted to investigate above the possibility in vivo, using the transgenic mice overexpressing ovine STAT5 (oSTAT5) under the control of MMTV promoter. Three transgenic mouse lines that expressed the exogenously introduced oSTAT5 in their mammary glands were established. The expression levels of exogenous oSTAT5 were higher than those of endogenous STAT in the mammary glands of all of the three transgenic lines. Although the expression levels of endogenous milk protein genes, WAP, and beta-casein genes, were not correlated with oSTAT5 expression, the expression levels of WAP and beta-casein were induced by exogenous oSTAT5 in the transgenic mice. The present study demonstrated, at very least, that STAT5 expression can directly activate the expression of milk protein genes, and particularly the WAP gene without PRL-mediated signal transduction.

Animals↗

Rat C peptide I and II stimulate glucose utilization in STZ-induced diabetic rats.

AIMS: To study the effects of physiological concentrations of rat proinsulin C peptide I and II, respectively, on whole body glucose utilization in streptozotocin diabetic and healthy rats. METHODS: A sequential insulin clamp procedure was used (insulin infusion rates 3.0 and 30.0 mU.kg-1.min-1) in awake animals. C-peptide infusion rates were 0.05 and 0.5 nmol.kg-1.min-1. Blood glucose was clamped at 7.7 +/- 0.3 mmol/l in the diabetic rats and at 3.9 +/- 0.1 mmol/l in the healthy rats. RESULTS: In diabetic rats infused at lower rates of C peptide and insulin, glucose utilization increased by 79-90% (p < 0.001) compared with diabetic animals infused with saline and insulin. Increasing the rate of C-peptide infusion tenfold did not elicit a statistically significant further increase in glucose utilization. C peptide I and II exerted similar effects. The metabolic clearance rate for glucose in the diabetic animals infused with C peptide was not different from that of the healthy rats. During high-dose insulin infusion (30.0 mU.kg-1.min-1) glucose utilization increased considerably and no statistically significant C-peptide effects were observed. About 85% of the increase in glucose utilization induced by C peptide could be blocked by treatment with N-monomethyl-L-arginine. CONCLUSIONS/INTERPRETATION: Physiological concentrations of homologous C peptide stimulate whole body glucose utilization in diabetic but not in healthy rats. C peptide I and II elicit similar effects. The influence of C peptide on glucose utilization may be mediated by nitric oxide.

Animals↗

Lung cancer in a child with a substantial family history of cancer.

The occurrence of primary lung cancer is rare in childhood. The case of an 11-year-old boy with primary lung cancer is presented in this report. He had a substantial family history of cancer. His chief complaint was coughing with right chest pain. A chest radiograph showed a coin lesion in the right lower lung. A right lower lobectomy revealed a squamous cell carcinoma (stage IIIA at Japanese TNM classification). Systemic chemotherapy using cisplatin, vindesine, THP-adriamycin and cyclophosphamide was performed. Six months after surgery, a recurrent tumor occurred. An analysis of the familial cancer related genes (p53 gene and mismatch repair gene) showed no abnormality.

Age of Onset↗

Cell-mediated immune responses in cattle vaccinated with a vaccinia virus recombinant expressing the nucleocapsid protein of rinderpest virus.

Rinderpest virus (RPV) is a member of the genus Morbillivirus in the family Paramyxoviridae which causes an acute and often fatal disease in large ruminants. To examine the immune response to the virus nucleocapsid (N) protein, a recombinant vaccinia virus expressing RPV nucleocapsid protein (rVV-RPV-N) was used to vaccinate cattle. The recombinant vaccine induced low levels of non-neutralizing anti-N antibodies. RPV-specific cell-mediated immunity induced by the recombinant was assessed by measuring both the lymphocyte proliferation and cytotoxic T-lymphocyte responses. The protective immune response was examined by challenging the vaccinated cattle with either a highly virulent (Saudi 1/81) or a mild (Kenya/eland/96) strain of the virus. The vaccinated cattle were not protected against challenge with the virulent RPV strain, except they showed a slight delay in the onset of disease when compared with the unvaccinated controls. In cattle challenged with the mild strain, apart from a transient fever, no clinical signs of rinderpest infection were seen in the vaccinated cattle. One out of two control cattle showed a similar response but the other died from classic rinderpest disease. Virus-neutralizing antibodies were induced more quickly following challenge with the mild strain in vaccinated cattle compared to the control animals. These data suggested that the cell-mediated immunity induced by rVV-RPV-N could stimulate the rapid production of neutralizing antibodies following RPV challenge but this response was not sufficient to protect against challenge with a virulent strain of the virus. Protection was seen in one of three animals challenged with a mild strain of the virus; however, a greater number of animals would need to be tested to estimate the significance of the protection afforded by the N protein.

Animals↗

Effect of direct application of estrogen aimed at lateral septum or dorsal raphe nucleus on lordosis behavior: regional and sexual differences in rats.

The role of estrogen in lordosis-inhibiting systems in the lateral septum or the dorsal raphe nucleus was investigated in female and male rats. Ovariectomized rats received implantation of 22-gauge guide cannulae to the bilateral or right side of the lateral septum (LS and rLS, respectively), the dorsal raphe nucleus (DRN) or bilateral cortex (CX). In castrated male rats, bilateral implantations of the cannulae to the LS were carried out (mLS). Three behavioral tests in total were performed at 2-week intervals. In the first test, all animals were subcutaneously injected with 1.5 microg/kg estradiol benzoate (EB). Forty-four hours after EB, 0.5 mg progesterone (P) was injected and a behavioral test was started 4 h after P. These hormonal regimes were used in all tests. In the second test, 2 h before EB injection, 27-gauge cannulae filled with estradiol (E2) were inserted into the DRN, LS or CX through the guide cannulae and were kept there for 4 h. In the third test, cholesterol was implanted instead of E2 into these areas. In the first test, most females showed low levels of lordosis quotient (LQ) and most males showed no lordosis. In the second test, mean LQs in the LS or rLS groups of females increased but not in the DRN and CX groups. In the mLS group no increase of LQ was observed. When cholesterol was implanted in the third test, mean LQs in all groups were as low as in the first test. These results suggest the possibility that estrogen releases the inhibition when it acts on the LS, but not on the DRN female rats. On the other hand, inhibition in the male LS may not be released by the direct action of estrogen.

Animals↗

Molecular cloning of cDNA for equine follistatin and its gene expression in the reproductive tissues of the mare.

A cDNA clone encoding equine follistatin was isolated from an equine ovarian cDNA library. Out of 1.2 x 10(5) independent clones screened, one positive clone was isolated and its cDNA sequence determined. The isolated clone, named EQ-FS-1, contained a complete open reading frame encoding 344 amino acid residues. The similarity of its deduced amino acid sequence to these of other mammalian species was greater than 95%. Although its expression level varied among the tissues examined, follistatin mRNA was detected in the equine uteroplacental tissues, follicles and corpora lutea by Northern blot analysis. In situ hybridization revealed that the expression of follistatin mRNA in the equine follicle was restricted exclusively to granulosa cells. When the expression pattern of follistatin mRNA in the equine uteroplacental tissues from mid- to late-pregnancy was examined, it was shown that its expression level tended to decrease after mid-pregnancy. These results suggest that follistatin acts in the reproductive tissues of the mare in maintaining pregnancy.

Amino Acid Sequence↗

Relationship between insulin and blood pressure in Japanese obese subjects.

OBJECT: The association of obesity and hypertension is well recognized. However, the nature of the relationship between increased body weight and blood pressure (BP) elevation has remained obscure. PATIENTS AND METHODS: We evaluated BP, insulin sensitivity, insulin clearance and fasting plasma insulin concentration in 19 younger (over 40 years) and in 15 older (more than 40 years) obese subjects to determine the relationships between BP and other factors. Insulin sensitivity and clearance were determined with the euglycemic clamp technique. RESULTS: BP was not associated with insulin sensitivity although most of the subjects showed insulin resistance. In the younger obese group, a positive correlation between diastolic BP and body mass index (kg/m2) was found (r=0.740; p=0.043). In the older obese group, systolic and diastolic BP were correlated with fasting plasma insulin levels (r=0.705; p=0.003; r=0.574; p=0.025, respectively), and systolic BP was inversely correlated with insulin clearance (r=-0.715, p=0.003). CONCLUSION: These results suggest that insulin is an important factor in BP elevation in older obese subjects, but not in younger obese subjects.

Adolescent↗

[Pulmonary metastatic malignant phyllodes tumor showing multiple thin walled cavities].

A 52-year-old woman who had undergone a partial mastectomy 1 year earlier because of benign phyllodes tumor was admitted because of dry cough and abnormal chest radiograph findings. Chest computed tomograms demonstrated multiple thin-walled cavities and nodules. Clinical examinations and transbronchial biopsy specimens failed to provide a conclusive diagnosis. However, the pulmonary thin-walled cavities enlarged, and a nodular shadow revealed cavitary formation. An open lung biopsy was performed to diagnose the pulmonary lesions. Although biopsy specimens disclosed the infiltration of poorly differentiated adenocarcinoma cells in pleura and pulmonary parenchyma, no primary site was detected. The patient did not respond to systemic chemotherapy (CDDP and VP-16), and died of respiratory failure due to advanced pulmonary metastasis. Autopsy demonstrated marked tumor invasion of the lungs, myocardium, and bone. We analyzed malignant cells in lung tissues at autopsy by immunohistochemistry, and found identical malignant cells in surgical samples obtained during the patients earlier mastectomy. A diagnosis of pulmonary metastasis from malignant phyllodes tumor of the breast was made. Thin walled cavitary lesions from malignant phyllodes tumor are rare; however, pulmonary metastasis of malignant phyllodes tumor should be considered one disease that exhibits thin-walled cavities as a radiographic manifestation.

Adenocarcinoma↗

Inhibitory effect of neural transections of dorsal raphe nucleus on induction of nocturnal prolactin surge by vaginal stimulation in ovariectomized rats.

The effect of complete (CC), anterior (AC) or posterior (PC) cut of the dorsal raphe nucleus (DRn) on induction of the nocturnal prolactin (PRL) surge by electrical vaginal stimulation (VS) was investigated in ovariectomized rats. Plasma level of PRL was measured by radioimmunoassay before and after VS. The data revealed that PRL levels increased in early morning on the day following VS in the rats without brain surgery or with sham-operation. In contrast, the nocturnal PRL surge did not occur in the CC, AC, or PC rats. These results suggest that both the anterior and the posterior fibers of the DRn plays an important role in induction of nocturnal PRL surge by VS in ovariectomized rats.

Analysis of Variance↗

Immunizing effect of vaccinia virus expressing the nucleoprotein of rinderpest virus on systemic rinderpest virus infection in rabbits.

A recombinant vaccinia virus (RVV) expressing the nucleoprotein (NP) of rinderpest virus (RPV) was examined in rabbits for the involvement of the NP protein in protection from the RPV infection. Despite their production of anti-NP antibody, the RVV-immunized rabbits succumbed to the RPV challenge, although there was a slight delay in the onset of disease after the low-dose challenge. On the other hand, the animals immunized with RVV expressing the hemagglutinin (H) protein of the RPV were completely protected. These results indicate that the NP protein might be not so effective as the H protein for the protection against viremic and systemic infection with RPV.

Animals↗

The effect of low intensity bicycle exercise on the insulin-induced glucose uptake in obese patients with type 2 diabetes.

OBJECTIVE: The present study was undertaken to reveal the effect of low intensity bicycle exercise on the insulin-induced glucose uptake in obese patients. SUBJECTS AND METHODS: Seven obese men with Type 2 diabetes (OBDM) and seven healthy young men (HY) participated in this study. The glucose infusion rate (GIR) was determined by glucose clamp procedure at an insulin infusion rate of 40 mU m-2 min-1 (plasma insulin concentrations: 700-800 pmol l-1). Confirming stabilized GIR, a 30-min bicycle exercise was performed during the glucose clamp which was continued for 120 min after exercise. RESULTS: Average GIR in OBDM for last 30 min prior to exercise were significantly lower than HY (28.3 +/- 1.7, 47.4 +/- 1.8 mumol kg-1 min-1 respectively, P < 0.05). GIR abruptly increased during exercise and gradually decreased after exercise to the nadir almost at the time from 30 to 60 min in recovery period in both groups. GIR in OBDM, however, gradually increased significantly over pre-exercise levels (P < 0.05), following exercise and reached the same levels compared to HY after 80 min of recovery period. CONCLUSION: These results indicated that in obese Type 2 diabetes, 30 min of low intensity bicycle exercise significantly enhances the lower level of insulin-induced glucose uptake shortly after exercise and might be useful for the treatment of post-prandial hyperglycemia.

Adult↗

Rapid analysis of allelic variants of the sheep PrP gene by oligonucleotide probes.

A rapid method to determine the allelic variants of the sheep PrP gene was developed. DNA samples from 128 Suffolk sheep (39 rams and 89 ewes) were screened by using polymerase chain reactions and dot-blot hybridization with 32P-labeled nine allele-specific oligonucleotide probes corresponding to the polymorphic PrP codons 112, 136, 154 and 171. Three allelic variants of the PrP gene, PrP(MARQ), PrP(TARQ) and PrP(MARR), were found in the flocks. Among those variants, nearly half of the ewes had alleles of the 171-Arg variant that is closely associated with resistance to natural scrapie. Assessments of allelic mutations of the PrP gene may help to select the scrapie-resistant progenitors in the flocks.

Alleles↗

Expression of inhibin alpha-subunit in horse testis.

Inhibin is believed to play roles in the pituitary secretion of FSH and in the paracrine regulation of testicular function. Although it has been generally accepted that inhibin is produced in Sertoli cells, there was a recent evidence for the localization of inhibin in Leydig cells of primates, rat and sheep. However, there is no report on the expression of inhibin in the adult horse testis. Therefore, using immunohistochemistry, western blotting and in situ hybridization techniques, the present study examined inhibin alpha-subunit (Ih-alpha) expression in the adult horse testis. For the detection of Ih-alpha protein, we used anti-porcine Ih-alpha antibody in immunohistochemistry and western blotting. Furthermore, digoxigenin-labeled complementary RNA probes were prepared to detect intracellular messenger RNA (mRNA) of Ih-alpha. Immunostainings for Ih-alpha were found not only in Leydig cells but also in Sertoli cells. The intensity in Leydig cells was stronger than in Sertoli cells. Immunoreactivities for Ih-alpha were found at approximately 46 kDa, 56 kDa and 90 kDa in the homogenates from testicular interstitial tissues. The bands at 56 kDa and 90 kDa agree with previous report, but not at 46 kDa. Signals for mRNA of Ih-alpha by in situ hybridization were detected in Leydig cells and in the basal region of seminiferous epithelium including Sertoli cells. These results suggest that Ih-alpha is expressed in Leydig cells and Sertoli cells of horse testis, and the expression level should be higher in Leydig cells than Sertoli cells.

Animals↗

Inhibitory effect of progesterone on androgen-induced lordosis in ovariectomized rats.

Androgen has a potency to induce female sexual behavior in ovariectomized rats. This study investigated the inhibitory effect of progesterone (P) on lordosis behavior facilitated by androgen. Ovariectomized female rats were given 100 to 800 micrograms/kg testosterone propionate (TP) or 5 micrograms/kg estradiol benzoate (EB). Forty-four hours after administration of the TP or the EB, all females received 0.5 mg P, and the first behavioral test was carried out four hours later. TP-treated animals showed lordosis responses in a dose dependent manner. Two weeks after the first test, the animals were retested by using the same hormonal regime as in the first test, except that an additional 5 mg P was administered 1 h prior to the TP or the EB. As a result, significant reductions in sexual receptivity were observed in the TP-treated as well as in the EB-treated females. These results indicate that progesterone can inhibit androgen-induced lordosis behavior in female rats.

Androgens↗

New approaches to the development of virus vaccines for veterinary use.

The marked progress in recombinant deoxyribonucleic acid (DNA) technology during the past decade has led to the development of a variety of safe new vaccine vectors which are capable of efficiently expressing foreign immunogens. These have been based on a variety of virus types--poxviruses, herpesviruses and adenoviruses--and have led to the production of many new potential recombinant vaccines. Of these recombinant vaccines, the rabies vaccine, in which the rabies G protein is expressed in a vaccinia vector, has been widely used in the field to prevent the spread of rabies both in Europe and in the United States of America. A recombinant Newcastle disease virus vaccine, using fowlpox virus as the vector to express immunogenic proteins from the Newcastle disease virus, has been licensed as the first commercial recombinant vectored vaccine. Many other recombinant virus vaccines are still at the stage of laboratory or field testing. The most recent breakthrough in vaccinology has been the success with the use of naked DNA as a means of vaccination. This approach has shown great promise in mouse model systems and has now become the most active field in new vaccine development. Molecular redesigning of conventional ribonucleic acid (RNA) viruses to obtain more stable attenuated vaccines was previously possible only for positive-strand RNA viruses, such as poliovirus. However, recent advances in molecular biological techniques have enabled the rescuing of negative-strand viruses from DNA copies of their genomes. This has made it possible to engineer specific changes in the genomes of Rhabdoviridae and Paramyxoviridae, both of which include several viruses of veterinary importance. The authors describe the current progress in the development of vector vaccines, DNA vaccines and vaccines based on engineered positive- and negative-strand RNA virus genomes, with special emphasis on their application to diseases of veterinary importance.

Adenoviridae↗

Wilms' tumor suppressor gene (WT1) as a target gene of SRY function in a mouse ES cell line transfected with SRY.

With the aim of identifying the gene(s) located downstream from SRY, we transfected an ES cell line with XX karyotype, TMA-18, with a Sry DNA construct and established cell lines, TS18-1 and TS18-2, where the transfected Sry was expressed in the functional linear mRNA form. Among the five potential SRY-target genes examined, i.e., MIS, SF1, P450arom, Sox9 and WT1, only the expression of WT1 was induced de novo by the unscheduled expression of Sry in the transfected cell lines. No clear indication of Sry-induced enhancement of Sox9 expression was obtained in the present series of experiments. Function of a yet unidentified gene(s) located on the Y chromosome might be needed for the up-regulation of Sox 9 expression which takes place during the development of male gonads. Quantitative RT-PCR analysis of the patterns of WT1 expression in developing fetal gonads revealed that although both male and female fetal gonads express WT1, male gonads invariably expressed WT1 mRNA at higher levels than female ones after the Sry expression. Immunohistochemical analysis of the male fetal gonads between 10.5 and 13.5 dpc demonstrated the presence of strong WT1 immunoreactivity in Sertoli cells of the primordial testes. Suggestions were made in the past indicating that both SF1 and WT1 proteins might be active in a common pathway upstream from Sry. Our results showed that WT1 is located downstream, rather than upstream from Sry and behaves independently from SF1. Analysis using an appropriate in vitro system will be essential to understand the molecular mechanisms of SRY action within cells.

Animals↗