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Biomedical subjects

H Hagiwara

Publications and source records attributed to H Hagiwara.

At least 91 records · Page 5Linked to original sources

Ultrastructural observation on 'transitional tubules' in human oviductal ciliogenic cells.

In the human oviduct epithelium during ciliogenesis, short tubular structures were found in the transitional zone between the basal body and cilium. The tubules, termed transitional tubules from their location, were 34-36 nm in diameter and 0.13 +/- 0.06 micron in length; the number around a basal body was variable, but usually 4-6. The cytoplasmic leaflets of the tubule membranes were coated by electron-dense material and appeared to be connected to alar sheets. The transitional tubules existed transiently during ciliogenesis. The exact role of transitional tubules is unknown, but considering their location, they may fix the basal body in the apical cytoplasm during ciliary elongation and/or may be related to formation of alar sheets.

Adult↗

Hepatitis C virus replicative levels and efficiency of genotyping by specific PCR and antibody assay.

We studied factors which influence the detection of hepatitis C virus genotypes by the group-specific PCR of the sequence within the core region gene and by the newly developed genotype-specific NS4 antibody assay. Genotyping was performed on 75 hepatitis C virus carriers in Japan, where patients with hepatitis C viremia are exclusively infected with genotype 1b, 2a, and 2b. PCR failed to identify genotypes in 8 (11%) patients, whereas 12 (16%) patients, including the 8 patients mentioned above, could not be genotyped by the serological assay. Serological genotypes showed almost complete agreement with those found by the PCR except that double infection was revealed in only two of the eight patients serologically judged to be coinfected with genotypes 1 and 2. In each assay, disease activity and levels of viremia assessed by a competitive reverse transcription PCR assay were significantly lower in patients infected with untypeable isolates than in those infected with typeable ones (P < 0.01). The PCR could identify the genotypes of isolates from all 64 patients with levels of viremia of > or = 10(6) copies/ml, and the genotype-specific antibody responses were found in 60 (94%) patients. In contrast, isolates from only 3 (27%) of 11 patients with low levels of viremia (< 10(6) copies/ml) could be genotyped by the PCR (P < 0.00001), and these patients showed the genotype-specific antibody responses (P < 0.00001). These findings suggest that low levels of hepatitis C virus replication may reduce the efficiency of genotyping by serological assay as well as by PCR.

Adult↗

Audiologic findings in patients with a point mutation at nucleotide 3,243 of mitochondrial DNA.

A mitochondrial tRNALeu(UUR) mutation at nucleotide 3,243 is known to be found in most patients with MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis, and strokelike episodes) and has also been identified in several families with maternally inherited diabetes mellitus and hearing loss. We report here audiologic features in patients with hearing loss associated with the mutation. Four patients without and five with MELAS were studied. Most of the patients had bilateral progressive sensorineural hearing loss. The most common shape of the audiogram was sloping, while cases in the advanced stages had flat audiograms. Speech discrimination scores were generally poor and did not parallel the degree of hearing loss. The present study suggests that the lesion for hearing loss could include both cochlear and retrocochlear involvement, but does not demonstrate a significant difference in the audiologic findings between patients with and without MELAS.

Adolescent↗

Gene expression of endothelial type isoform of nitric oxide synthase in various tissues of stroke-prone spontaneously hypertensive rats.

Nitric oxide (NO) has been suggested to play important roles in the pathophysiology of various cardiovascular diseases. This study tested the hypothesis that an attenuated biological action of NO in hypertension is attributed to a change in the gene expression of NO synthase (NOS), a key enzyme involved in NO formation. The expression level of mRNA of endothelial type NOS (NOS-III) was determined in stroke-prone spontaneously hypertensive rats (SHR-SP/Izm) and Wistar Kyoto rats (WKY/Izm) by ribonuclease protection assay using a partial clone as probe. NOS-III mRNA was expressed ubiquitously in various tissues of WKY/Izm and SHR-SP/Izm either at 5 wk or 13 wk of age. There was no significant difference in the tissue expression of NOS-III mRNA between the two strains at either age. The intensity and localization of the hybridization signal for NOS-III mRNA in the heart of SHR-SP/Izm did not differ from those in the heart of WKY/Izm. These results suggest that the attenuated biological action of NO implied in genetically hypertensive rats is not attributed to an abnormality at the level of NOS-III mRNA expression in the tissues, although lack of an increase in NOS-III gene expression, despite the hypertensive hemodynamic stress, may modify the blood pressure in hypertension.

Animals↗

High levels of serum interleukin-10 are associated with a poor response to interferon treatment in patients with chronic hepatitis C.

BACKGROUND: It still remains unclear whether some immunologic factors affect the response to interferon treatment. We therefore examined whether the pretreatment levels of serum interleukin-10 and soluble intercellular adhesion molecule-1 can be associated with the response to interferon treatment in patients with chronic hepatitis C. METHODS: One hundred and two patients with chronic hepatitis C treated with interferon alpha-2b were divided into three groups on the basis of patterns of biochemical interferon response. Pretreatment levels of serum interleukin-10 and soluble intercellular adhesion molecule-1 were determined using enzyme-linked immunosorbent assay. Hepatitis C virus (HCV) typing was performed with a serologic enzyme-linked immunosorbent assay. RESULTS: For patients with serotype I (n = 76) the numbers of sustained, transient, and non-responders were 12 (16%), 43 (56%), and 21 (28%), respectively. In serotype-I patients the pretreatment levels of serum interleukin-10 in non-responders were significantly higher than those in sustained or transient responders, although no significant differences were observed in HCV RNA quantity between them. There were no significant differences in the pretreatment levels of serum soluble intercellular adhesion molecule-1 among the three groups. CONCLUSION: These findings suggest that high serum interleukin-10 levels may be related to a poor response to interferon treatment in serotype-I patients with chronic hepatitis C.

Adult↗

[A study on the quantitative evaluation method of human alertness and its application].

This article presents an alertness evaluation method being analyzed by the authors, and discusses its application. With the aim of quantifying the physiological activity of the living body, the alpha-attenuation test (AAT) for evaluation of alertness was assessed. AAT is a method of quantitatively determining alertness level from alpha-wave power spectra during eye closure and opening. 1. The potential for quantification of diurnal change of alertness by the use of the AAT is suggested. Also rated, are a performance evaluation method based on reaction time and target tracking error and a subjective feeling evaluation method using an analog scale on a computer display. Using these methods of evaluation, measurements were made in 9 healthy men and women at 2-hour intervals over the period from 09: 30 to 19: 30. The findings are summarized as follows: 1) Alertness lowered from 13: 30 to 15: 30, as rated by diurnal change determined by the AAT 2) The performance test and subjective feeling scoring revealed decreased target tracing ability and extended reaction time from 13: 30 to 15: 30, accompanied by reduced performance, increased sleepiness, and failure to achieve accurate, quick action. 2. The relation between the quality of rest and the alpha-attenuation coefficient (AAC), the quantitative index of the AAT, was rated. There were some statistical significances between the quality of rest and the AAC. These results suggest that the AAC is found applicable to the quantitative assessment of diurnal changes of physiological activity, and that the AAC is reflected in the quality of rest.

Adult↗

Circadian rhythm estimation by core body temperature filtered with simultaneously recorded physiological data.

In field measurements, monitoring of core body temperature is influenced by physical activities; therefore, the estimation of circadian rhythm from the data may not be exact. The purpose of this study is to design a core body temperature filter in order to reduce artifacts induced by physical activities using simultaneously recorded physiological data such as heart rate data. The effects of physical activities on core body temperature and heart rate are assessed through three experiments. Based on the above knowledge, a core body temperature filter was designed. The filter removes part of rectal temperature data as artifact when heart rate rises above a predetermined threshold. As a result, most of the spike-like noise was removed and the filtered temperature data showed sinusoidal variation more than the unfiltered data. The mesor of the estimated rhythm significantly decreased. This filtering method can provide more precise information about circadian rhythm, especially in field measurements.

Adult↗

Stimulation by retinoids of the natriuretic peptide system of osteoblastic MC3T3-E1 cells.

The effects were examined of treatment with retinoids of osteoblastic MC3T3-E1 cells on the natriuretic peptide system that promotes the differentiation of osteoblastic cells. Northern blot analysis revealed high levels of mRNA for the retinoid X receptor beta (RXR beta) and moderate levels of mRNAs for retinoic acid receptors alpha (RAR alpha) and gamma (RAR gamma). Exposure of MC3T3-E1 cells to 1 microM retinoid caused increases in the levels of C-type natriuretic peptide (CNP) and natriuretic peptide receptor-C (NPR-C). The activity of natriuretic peptide receptor-B (NPR-B) was unchanged after the addition of retinoid to the culture system. These results suggest that retinoids might influence the metabolism of osteoblastic cells through regulation of the natriuretic peptide system.

Animals↗

A Case of Ductal Carcinoma In Situ (DCIS:noncomedo type)Detected by Ultrasonography: Demonstration of Occult Multiple Foci.

We report a case of ductal carcinoma in situ (DCIS) of the breast detected by ultrasonographic mass screening in a 51-year-old woman. In a mass screening program for breast cancer, physical examination with inspection and palpation, and ultrasonography (US) were performed. A hypoechoic mass with a slightly irregular margin was detected by US in the lateral upper quadrant of the right breast, at a distance 2 cm from the edge of the nipple. The mass was not detected by physical examinations or by mammography (MMG). The mass, which measured 0.8 x 0.5 cm and was examined by fine needle aspiration biopsy (FNAB) under US guidance, was cytologically diagnosed as class X. Modified radical mastectomy (Auchincloss method) was performed with the patient's consent. Pathological examination of the resected specimen revealed DCIS (noncomedo type) and occult multiple foci of malignancy which was considered tracking centripetally underneath the nipple. This case suggests that US and FNAB performed under US guidance are useful in the detection and diagnosis, respectively, of a breast mass. We should take multifocality into consideration, particularly with tendency tracking to the nipple, in the treatment of small breast cancers such as DCIS.

Journal Article↗

Inhibition of proliferation of chondrocytes by specific receptors in response to retinoids.

All-trans retinoic acid inhibited the proliferation of chondrocytes derived from rat xiphoid cartilage when added to the culture medium at 10(-10)-10(-8) M. Proliferation of mouse clonal osteoblastic cells was also inhibited to a significant extent by all-trans retinoic acid. However, no such inhibitory effects on rat smooth muscle cells and human fibroblasts were observed. Flow cytometric analyses of chondrocytes labeled with propidium iodide revealed that all-trans retinoic acid arrested chondrocytes at the G1 phase of the cell cycle. Since 9-cis retinoic acid, which is synthesized enzymatically from all-trans retinoic acid, also inhibited the proliferation of chondrocytes, we investigated the subtypes of retinoic acid receptors in chondrocytes. Northern blot analysis revealed high levels of mRNA for retinoid X receptor alpha (RXR alpha), moderate levels of mRNA for retinoic acid receptor gamma (RAR gamma), and low levels of mRNA for RAR alpha and RXR beta. The mRNA for RAR beta and RXR gamma were not detected. These results suggest that retinoids are associated with the inhibition of proliferation of chondrocytes via both families of nuclear receptors, namely, RARs and RXRs.

Animals↗

Stimulation by C-type natriuretic peptide of the differentiation of clonal osteoblastic MC3T3-E1 cells.

We examined the effects of C-type natriuretic peptide (CNP) and B-type natriuretic peptide receptor (NPR-B) system, which stimulates the intracellular production of cGMP, on osteoblastic differentiation using clonal murine calvarial MC3T3-E1 cells. CNP-like immunoreactivity was detected in the conditioned medium and in lysates of MC3T3-E1 cells. Exposure of cells to CNP caused an increase in the intracellular production of cGMP and the increase was dose-dependent, while ANP had no effect. These results imply that CNP regulates osteoblastic metabolism via NPR-B in an autocrine manner. Northern blot analysis revealed that treatment of MC3T3-E1 cells with CNP increased the steady-state levels of mRNAs for type-I collagen, cellular alkaline phosphatase (ALPase), and osteocalcin, which are well known as markers of osteoblastic differentiation. Our observations suggest the possibility that CNP functions as a local regulator of osteoblastic differentiation, acting via a cGMP-mediated pathway.

3T3 Cells↗

In situ identification of messenger RNA of endothelial type nitric oxide synthase in rat cardiac myocytes.

The endothelial type nitric oxide synthase (NOS) termed as NOS-III has been demonstrated in various nonendothelial cells. Although NOS-III was recently found in the cardiac tissues, identification of the cells expressing NOS-III gene in the heart remains unknown. In this study, we compared the expression of NOS-III mRNA in various tissues of rats by Northern blot analysis using a homologous cDNA probe and identified the cell type by in situ hybridization histochemistry with a cRNA probe. NOS-III mRNA was found more abundantly in atria and ventricle than in other various tissues examined. NOS-III mRNA was intensely and specifically visualized in the cardiac myocytes as well as in the endothelial cells of the heart. No staining was seen with a sense cRNA probe. These results provide the first in vivo evidence for NOS-III mRNA in the cardiac myocytes and suggest its role in the regulation of cardiac functions.

Animals↗

Angiotensin converting enzyme inhibitor normalizes vascular natriuretic peptide type A receptor gene expression via bradykinin-dependent mechanism in hypertensive rats.

We previously demonstrated that angiotensin converting enzyme (ACE) inhibitor normalizes the up-regulated gene expression of vascular natriuretic peptide type A (NP-A) receptor in hypertensive rats. To elucidate the mechanism, we examined the effect of angiotensin II receptor (AT1) antagonist (TCV-116) and bradykinin receptor (B2) antagonist (Hoe 140) on the NP-A receptor mRNA level in the aorta of genetically hypertensive rats (SHR-SP/Izm) using ribonuclease protection assay. The effect of ACE inhibitor on the NP-A receptor mRNA level was completely abolished by a concomitant administration of Hoe 140, while TCV-116 did not show any significant effect on the NP-A receptor mRNA level. These results suggest that bradykinin plays an important role in the regulation of the vascular NP-A receptor gene expression.

Angiotensin-Converting Enzyme Inhibitors↗

Sequencing of a 23 kb fragment from Saccharomyces cerevisiae chromosome VI.

Plasmid clone gapB and lambda phage clone 4682, which contain fragments of Saccharomyces cerevisiae chromosome VI, were analysed. A 23 kb sequence was determined and ten open reading frames (ORFs) were revealed. Among them, five ORFs were identical to five yeast genes (SEC4, MSH4, SPB4, DEG1 and NIC96), two were identical to transposable elements (TYA and TYB), one (gapBorfF003) was highly homologous to a yeast expressed sequence tag, and another (4682orfF002) was predicted to be a nuclear protein. Sequence data have been submitted to DDBJ/EMBL/GenBank data library under Accession Number D44604 (clone gapB) and D44600 (clone 4682), respectively.

Amino Acid Sequence↗

Analysis of a 36.2 kb DNA sequence including the right telomere of chromosome VI from Saccharomyces cerevisiae.

The nucleotide sequence of a 36.2-kb distal region containing the right telomere of chromosome VI was determined. Both strands of DNA cloned into cosmid clone 9965 and plasmid clone pEL174P2 were sequenced with an average redundancy of 7.9 per base pair, by both dye primer and dye terminator cycle sequencing methods. The G+C content of the sequence was found to be 37.9%. Eighteen open reading frames (ORFs) longer than 100 amino acids were detected. Four of these ORFs (9965orfR017, 9965orfF016, 9965orfR009 and 9965orfF003) were found to encode previously identified genes (YMR31, PRE4, NIN1 and HXK1, respectively). Six ORFs (9965orfR013, 9965orfF018, 9965orfF006, 9965orfR014, 9965orfF013 and 9965orfR020) were found to be homologous to hypothetical 121.4-kDa protein in the BCK 5' region, Bacillus subtilis DnaJ protein, hypothetical Trp-Asp repeats containing protein in DBP3-MRPL27, putative mitochondrial carrier YBR291C protein, Salmonella typhimurium nicotinate-nucleotide pyrophosphorylase, and Escherichia coli cystathionine beta-lyase, respectively. The putative proteins encoded by 9965orfF018, 9965orfR014 and 9965orfR020 were found to be, respectively, a new member of the family of DnaJ-like proteins, the mitochondrial carrier protein and cystathionine lyase.

Amino Acid Sequence↗

Fifteen open reading frames in a 30.8 kb region of the right arm of chromosome VI from Saccharomyces cerevisiae.

The nucleotide sequence of cosmid clone 9765, which contains 30.8 kb of the right arm of chromosome VI, was determined. Both strands were sequenced, with an average redundancy of 8.17 per base pair by both dye primer and dye terminator cycle sequencing methods. The G+C content of the sequence was found to be 40.3%. Fifteen open reading frames (ORFs) greater than 100 amino acids and one tRNA-Tyr gene (SUP6) were detected. Seven of the ORFs were found to encode previously identified genes (HIS2, CDC14, MET10, SMC2, QCR6, PH04 and CDC26). One ORF, 9765orfF010, was found to encode a new member of the Snf2/Rad54 helicase family. Three ORFs (9765orfR002, 9765orfR011 and 9765orfR013) were found to be homologous with Schizosaccharomyces pombe polyadenylate binding protein, Escherichia coli hypothetical 38.1-kDa protein in the BCR 5' region, and transcription regulatory protein Swi3, respectively.

Adenosine Triphosphatases↗

Long-term biochemical and virological response to natural interferon-alpha in patients with chronic hepatitis C.

To determine the long-term response to interferon-alpha, 134 patients with chronic hepatitis C were followed for more than a year after therapy. Follow-up was stopped for 14 patients and 43 patients received retreatment. The remaining 77 patients were followed for 26-46 months, and 39 of them achieved long-term sustained alanine aminotransferase (ALT) normality. This normality was achieved in 35/38 short-term sustained responders, which was significantly higher (P < 0.001) than in the short-term relapsers (2/38) and the short-term nonresponders (2/44). Hepatitis C virus (HCV) RNA remained negative in 27 (73%) of 37 patients in the long-term sustained ALT normality group. However, only one (1.6%) of 63 long-term responders was negative for HCV RNA. The results of HCV RNA testing for the long-term period agreed with those at three to six months after therapy. Serum ALT levels during the first six months after therapy and HCV RNA testing at three to six months after therapy are important for predicting long-term sustained ALT normality and HCV RNA negativity.

Adult↗