Biomedical subjects

H Sakai

Publications and source records attributed to H Sakai.

At least 595 records · Page 33Linked to original sources

PubMed1995

Characterization of rat N-acetylglucosaminyltransferase I expressed in Escherichia coli.

The catalytic domain (sGnT-I) of rat liver N-acetylglucosaminyl-transferase I (GnT-I) was expressed in Escherichia coli. Lysates from pETsGnT-I transformants contained a prominent protein species of 46 kDa with which a significant GnT-I activity was associated. To purify the relevant enzyme, we constructed cDNAs encoding sGnT-ICH and sGnT-INH, which had six additional histidine residues as an affinity tag at the C-terminal and the N-terminal of sGnT-I, respectively, and introduced them into E. coli cells for expression. sGnT-INH was purified and its enzymatic properties were examined.

Animals↗

PubMed1995

Reddish Escherichia coli cells caused by overproduction of Bacillus stearothermophilus uroporphyrinogen III methylase: cloning, sequencing, and expression of the gene.

During shotgun cloning of an amylase gene, we found a transformant of Escherichia coli with a reddish color. The transformant produced highly water-soluble red pigments the molecular masses of which were less than 3000. The plasmid harbored by the transformant contained a DNA fragment derived from a strain of Bacillus stearothermophilus. Truncation of the insert DNA showed that an 1.1-kbp Sau 3A-SalI fragment was responsible for the reddish colony. An open reading frame was found in the nucleotide sequence of the 1.1-kbp DNA fragment. The production of the red pigment was accompanied by a colorless 28-kDa protein. The sequence of the 28-kDa protein was highly homologous to bacterial uroporphyrinogen III methylases participating in corrinoid biosynthesis. The 28-kDa protein was found to be a thermostable uroporphyrinogen III methylase.

Amino Acid Sequence↗

PubMed1995

Tissue- and development-specific expression of goat insulin-like growth factor-I (IGF-I) mRNAs.

We cloned four kinds of goat insulin-like growth factor-I (IGF-I) cDNAs. Each of them encodes the same mature IGF-I and different signal peptides. In this study, we analyzed expression of the four kinds of mRNAs in tissues in various developmental stages by a reverse transcriptase-polymerase chain reaction assay. The results suggest that their expression may be controlled by different promoters, the activities of which are tissue- and development-specific.

Animals↗

PubMed1995

Dynamic aspects in the expression of the goat insulin-like growth factor-I (IGF-I) gene: diversity in transcription and post-transcription.

Goat IGF-I (gIGF-I) cDNA was cloned using the reverse transcriptase-polymerase chain reaction (RT-PCR). The cDNA, which was homologous to rat class 1 IGF-I cDNA, was 969 bp long. From the open reading frame found in it, we predicted a 154 amino acid protein consisting of a 49 amino acid signal peptide, a 70 amino acid mature IGF-I peptide, and a 35 amino acid E domain (the COOH-terminal peptide). From the gIGF-I gene, we isolated and sequenced some segments containing four exons that encompassed the entire gIGF-I cDNA sequence. Goat liver RNA was analyzed by RT-PCR, and the nucleotides of the RT-PCR products were sequenced and checked with the nucleotide sequences of the segments from the gIGF-I gene. The gene had three leader exons (1W, 1, and 2, from upstream to downstream) and was transcribed into three kinds of mRNAs (classes 1W, 1, and 2). Another RNA species was detected by RT-PCR analysis of exon 1W. We sequenced it and found that in this transcript, the 3'-portion of exon 1 was inserted between exons 1W and 3, resulting in class 1W-1 del. mRNA. That is to say, the gIGF-I gene had three leader exons and four kinds of mature mRNA.

Amino Acid Sequence↗

PubMed1995

Functional distinction among structural subsections in the specific priming signal for DNA replication of the broad host-range plasmid RSF1010.

To analyze the functional contribution to the ssiA function of subsections of the ssiA-determinant sequence based on their dimensions, we constructed ssiA mutants carrying insertions and deletions. Results of the examination of the ssiA mutants told us that, in addition to the base sequence, the dimensions were crucial factors for the functional contribution of the subsections of ssiA.

Bacterial Proteins↗

PubMed1995

Immunolocalization of CD44 and heparan sulfate chains on the stratum intermedium and papillary layer in the rat enamel organ.

We studied the immunohistochemical localization of CD44 and heparan sulfate (HS) chains in rat enamel organ by confocal laser scanning microscopy and transmission electron microscopy. We also investigated the binding sites of basic fibroblast growth factor (bFGF), one of the heparin-binding growth factors (HBGF), on Microslicer-sections to clarify its role in the cell-cell interaction of HS. At the differentiation stage of ameloblast, weak immunoreactivity for CD44 was detected on the plasma membrane of the inner enamel epithelium, external enamel epithelium and the cells adjacent to the inner enamel epithelium. In accordance with the differentiation of preameloblasts into secretory ameloblasts, this immunoreactivity increased in stratum intermedium cells. At the secretory stage, stratum intermedium cells showed the most intense immunoreactivity in the enamel organ. At the maturation stage, strong immunoreactivity was seen on papillary layer cells. On the other hand, the lateral plasma membrane of ruffle-ended (RA) and smooth-ended ameloblast (SA) showed weak reactivity. No immunoreactivity was detected on the ruffled border of RA and the distal plasma membrane of SA. Immunolocalization of HS chains was similar to that of CD44. The binding activity of bFGF was also intense on stratum intermedium cells and papillary layer cells. These findings suggest that: 1) stratum intermedium cells and papillary layer cells express CD44 and HS chains in accordance with their differentiation; 2) HS chains on the plasma membrane of these cells may regulate calcium transport by their negative charge; and 3) HS chains on the stratum intermedium and papillary layer may play an important role in the differentiation and activity of ameloblasts by preserving HBGF.

Animals↗

PubMed1995

Intravenous single and repeated dose toxicity studies of cimadronate (YM175), a novel bisphosphonate, in rats.

Cimadronate (YM175) is a novel bisphosphonate with potent inhibitory activity on bone resorption under development for the treatment of tumor-induced hypercalcemia, metastatic bone disease and osteoporosis. We conducted toxicity studies of cimadronate intravenously, single dose, 30-day repeated dose and 26-week weekly dose in F344 rats. In the single dose study, cimadronate was administered to rats and the animals were observed for 14 days. Major toxic symptoms were decreased motility and piloerection and LD50 values were 23 mg/kg for males and 21 mg/kg for females. In the 30-day study, the animals received cimadronate at doses of 0, 0.16, 0.31, 0.62 or 1.25 mg/kg/day. At 0.16 mg/kg/day or more, an increased amount of primary spongiosa was observed in the femur. At 0.62 mg/kg/day or more, renal and testicular/epididymal toxicity were observed. After a 30-day recovery period, the finding in the kidney disappeared, but the findings remained in the bone, testis and epididymis. In the 26-week weekly dose study, animals received cimadronate at doses of 0, 0.31, 0.62 and 1.25 mg/kg/week. At 0.31 mg/kg/week or more, an increased amount of primary spongiosa was seen in the femur. Renal and testicular/epididymal toxicity, however, were not observed.

Animals↗

PubMed1995

Urinary albumin fragments as a new clinical parameter for the early detection of diabetic nephropathy.

Urinary albumin fragments (uAF) from patients with NIDDM were analyzed as a possible factor in the early discovery of diabetic nephropathy before emergence of microalbuminuria. SDS-PAGE and immunoblot assay were employed for detection of uAF. Samples from 252 patients with NIDDM, 158 patients with non-diabetic diseases, and 48 healthy volunteers were examined; uAF were detected in 139 (55.2%), 94 (59.5%), only one (2.1%), respectively. In diabetic patients with normoalbuminuria, uAF were detected in 48 out of 159 cases (30.2%). Two years after the initial study, 3 of the 17 diabetic patients (17.6%) with normoalbuminuria and uAF showed micro- or macroalbuminuria. It was concluded that detection of uAF might be useful for the early detection of diabetic nephropathy in NIDDM.

Adolescent↗

PubMed1995

GTP-binding protein-mediated production of superoxide anion in rabbit gastric parietal cells.

We found that guanosine-5'-O-(3-thiotriphosphate) (GTP gamma S) increased superoxide production in the suspension rich in rabbit gastric parietal cells that were permeabilized by streptolysin O. In this experiment, oxidation of non-fluorescent dihydrorhodamine 123 into the fluorescent rhodamine 123 by oxygen radicals was measured. Results support that GTP gamma S closes a housekeeping C1- channel via intracellular production of superoxide anion in the parietal cell.

Animals↗

PubMed1995

[Case report: rheumatoid arthritis complicated by chronic myelogenous leukemia treated with interferon].

A fifty-six-year old man complained of arthralgia and swelling of both feet, morning stiffness in both hands and finger joints in March 1987, and was treated with non-steroidal anti-inflammatory agents at another hospital. He has been treated for chronic myelogenous leukemia (CML) since May 1990. He was admitted to our hospital in March 1991 because of worsening of his multiple arthralgias, and a diagnosis of rheumatoid arthritis (RA) (Stage I, Class 2) was made on the basis of gait disturbance, arthralgia persisting for more than 6 weeks, the presence of subcutaneous nodules and X-ray findings. CML was confirmed by peripheral blood and bone marrow findings and the presence of the Philadelphia chromosome and bcr gene rearrangement. High fever and dyspnea developed suddenly 3 days after administration of interferon in May 1991. In addition to pneumonia, a leucostasis was suspected and he was treated with high dose steroids and antibiotics. After improvement, the steroids were tapered and he was discharged from hospital in July 1991.

Antineoplastic Agents↗

PubMed1995

In situ hybridization of interleukin 6 in diabetic nephropathy.

Increased mesangial expansion is one of the most characteristic histological changes in diabetic nephropathy (DN). Although the pathogenesis of DN remains unclear, recent studies associate interleukin (IL) 6 with mesangial proliferative glomerulonephritis. To elucidate the expression and localization of IL-6 mRNA in renal tissues of patients with DN, a high-resolution in situ hybridization using digoxigenin-labeled oligonucleotide was performed. Patients were divided into three groups based on light microscopy findings: mild (group 1), moderate (group 2), and severe (group 3) mesangial expansion. The relationship between the expression of IL-6 mRNA and the degree of glomerular mesangial expansion in DN was examined. Individual cells positive for IL-6 mRNA were observed in glomeruli. These cells were mesangial cells, glomerular epithelial cells, and Bowman's capsule. The signal intensity was strongest in tissues from group 2 but was weak in those from groups 1 and 3. Most cells in the area of mesangial proliferation were strongly stained for IL-6 mRNA, and few positive cells were found in the Kimmelstiel-Wilson nodular lesion. In the interstitium, some tubules, particularly atrophic tubules, and some infiltrating cells were positively stained for IL-6 mRNA. The interstitial expression of IL-6 mRNA correlated significantly with the degree of interstitial injury and was remarkable in tissues from groups 2 and 3. We conclude that IL-6 mRNA is expressed by glomerular resident cells and interstitial cells in the renal tissue of patients with DN and that its expression may be associated with mesangial proliferation and may be involved in the tissue injury of DN.

Adolescent↗

PubMed1995

Effect of an alpha 2-adrenoceptor antagonist, midaglizole, on bronchial responsiveness to histamine in patients with mild asthma.

The purpose of this study was to investigate the effects of midaglizole (MID) and interactions of MID with beta 2-agonists on bronchial hyperresponsiveness (BHR). Thirteen patients with mild asthma were randomized with respect to whether they were taking oral beta 2-agonists, and the investigators were blinded to patients' regular drug regimens. Histamine challenge test was performed before and after oral administration of MID for 1 week (300 mg/day). There was a significant increase in PC20 values in asthmatic subjects on regular therapy with beta 2-agonists (p < 0.05), whereas asthmatics not taking beta 2-agonists showed no significant change in PC20. These results suggest that MID may participate in improved BHR in the presence of beta 2-agonists and play some role in enhancing the effects of beta 2-agonists.

Administration, Oral↗

PubMed1995

Protective effect of an alpha 2-adrenoceptor antagonist, midaglizole, against allergen-provoked late asthmatic responses.

We investigated the effects of midaglizole (MID) on the allergen-provoked early and late asthmatic response (EAR, LAR) in a randomized, single-blind protocol. Four subjects with allergic asthma inhaled the following combinations: (1) allergen alone (control phase); (2) (a) pre- and posttreatment with MID (oral administration of 200 mg), (b) pretreatment with MID (200 mg), or (c) posttreatment with MID (200 mg). MID significantly inhibited the LAR in the three treatment phases (p < 0.05). However, the EAR was not inhibited significantly. These results suggest that MID may participate in the inhibition of the allergen-provoked LAR and may play an important clinical role as an anti-LAR agent.

Adrenergic alpha-Antagonists↗

PubMed1995

[[The study of socio-demographic behavior relevant to "Hinoe-uma" in 1966]].

Aspects of the fall in the number of births occurring in Japan in 1966 are explored. "The studies of 'Hinoe-uma' seem to have lacked...three perspectives; (1) the scope of demographic influence of 'Hinoe-uma', (2) demographic characteristics from the viewpoint of children who were born in the year 'Hinoe-uma', (3) socio-demographic influence of 'Hinoe-uma'. So we explored the various data regarding the three points in 1966 'Hinoe-uma'.... We confirmed by macro data that both Koreans and Chinese in Japan, Japanese in foreign countries and parents who had [an] illegitimate child in 'Hinoe-uma' were also influenced by superstitions [about] 'Hinoe-uma'." (SUMMARY IN ENG)

Asia↗

PubMed1995

[Frontal acute extradural hematoma due to contrecoup injury: a case report].

Acute extradural hematoma, a collection of blood between the inner table of the skull and the dura, generally occurs in coup craniocerebral trauma with skull fracture. We report here a case of a 52-year-old female who had had a traffic accident. She suffered a serious wound consisting of a subcutaneous hematoma with skull fracture and so called Battle's sign on the left occipital region but not wounded in any other region. CT scan obtained shortly after the injury demonstrated a thin acute extradural hematoma with intracranial air under the subcutaneous hematoma in the left occipital region, and another hematoma with subarachnoid hemorrhage due to contrecoup head trauma in the right frontal region. We treated her conservatively with a common drip. She sometimes vomited during the several hours after admission. CT scan six hours later showed that the contrecoup extradural hematoma had enlarged. Immediately, we carried out the evacuation of the hematoma with decompressive craniectomy. Her scalp, cranium and frontal skull base over the extradural hematoma were without injury, and multiple small haemorrhages were found to have occurred on the surface of the dura that had been separated from the inner table of the skull. After the operation, the patient recovered consciousness. Contrecoup acute extradural hematoma is very rare. It seems that the appearance of hematoma in our case resulted from the frontal dural separation due to distortion of the cranium brought on by the force of the impact and the subsequent gradual growth of the hematoma under the stimulation of several bouts of vomiting.

Acute Disease↗

PubMed1995

[Development of quality of life (QOL) questionnaire for use of lung cancer patients in palliative therapy--study of validity and reliability. No.1].

We investigated the validity and reliability of QOL questionnaire for lung cancer patients in palliative therapy. The questionnaire covered twelve items: appetite, feelings, sleep, mental and physical fatigue, pain, anxiety, daily activity, abdominal and respiratory conditions, linear and face scales. The data were collected from 65 patients and analyzed with principal component analysis and correlation analysis. 1) The percentage of complete answers was 81.5%. 2) Appetite, feelings, sleep, mental fatigue, anxiety and mental scale, pain, respiratory condition, abdominal condition, physical fatigue and physical scale, and satisfied internal consistency. 3) The test-retest reliability was satisfied 4) The inquiry items were grouped into physical, mental and activity scales, and these scales belonged to different dimension. 5) There were correlations between a linear scale, face scale, total score and items. 6) In concurrent validity, there were correlations between performance status and the activity scale, SDS, STAI and the mental scale. 7) In sensitivity, the total score and face score were worst within one week after chemotherapy, and then recovered. This questionnaire was indeed valid and reliable for use as a QOL questionnaire for lung cancer patients in palliative therapy.

Adult↗

PubMed1995

[Development of quality of life (QOL) questionnaire for use of lung cancer patients in palliative therapy--study of validity and reliability no. 2, the effects of chemotherapeutics in QOL].

The effects of cisplatin and carboplatin in QOL were studied using a QOL questionnaire seeking to conform their validity and reliability in lung cancer patients. The questionnaire was composed of eleven items; appetite, feeling, sleep, mental fatigue, pain, anxiety, daily activity, abdominal and respiratory conditions, linear analog and face scale as global scale. The data were collected from 21 patients treated with cisplatin (Cis group) and 9 patients administered carboplatin (Carbo group). Chronological changes of QOL were measured by AUC (area under the curve) method. 1) The total score of 9 items, linear analog and face scales rose immediately to the highest levels (worse) after treatment and maintained this level for 1 week in the Cis group. The Carbo group levels rose from 3 days and returned to the control level at 9 days after treatment. 2) AUC of the total score, linear analog and face scales in the Cis group increased significantly when compared with those of the Carbo group. 3) When compared with the Cis and Carbo groups the physiological and active scales were not different, but the psychological scale showed a significant difference between the two groups. 4) The total score of the psychological scale correlated the abdominal score in the Cis group, but not in the Carbo group. 5) Sleep and mental fatigue were related to the aggravation of QOL at 5-6 days after Carboplatin treatment. These results suggested that this QOL questionnaire had sufficient sensitivity to reflect any chemotherapeutic side-effects. 6) AUC is useful method in chronological evaluation of QOL.

Adult↗

PubMed1995

A clinical study of mortality due to asthma.

BACKGROUND: Despite identification of the pathophysiologic mechanisms of asthma and improvement in therapy, asthma mortality has not decreased in recent years. OBJECTIVE: The pathophysiology and asthma-related death preventive measures were investigated with a physician-based questionnaire survey. METHODS: Questionnaires were sent to physicians primarily involved in treating asthma in Nagasaki Prefecture, Japan. The clinical characteristics of 32 patients who died of asthma (fatal cases) from 1984 to 1992 were compared with those of 17 patients with severe asthma who survived as a result of treatment by mechanical ventilation (nearly fatal cases). RESULTS: The number of deaths due to asthma increased in the last 2 years. Fatal cases and nearly fatal cases included patients with severe asthma as well as patients with mild asthma. Analysis of the clinical histories of patients judged to have died suddenly revealed the presence of persistent wheezing in these patients for a few days prior to the fatal episode. Airway obstruction was more marked and bronchial hyperresponsiveness was greater in fatal cases compared with those of a group of 70 patients without a history of nearly fatal acute asthma. CONCLUSIONS: The following measures may help prevent asthma deaths. Both patients and physicians should realize that even mild episodes can lead to severe, even fatal acute asthma. The severity of asthma should be evaluated not only by symptoms and peak expiratory flow rates but also bronchial hyperresponsiveness. Treatment should include reduction of bronchial hyperresponsiveness using oral or inhaled corticosteroids.

Adult↗