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Biomedical subjects

F Sakai

Publications and source records attributed to F Sakai.

At least 55 records · Page 3Linked to original sources

The effects of hypothermia on a cloned human brain glutamate transporter (hGLT-1) expressed in Chinese hamster ovary cells: -[3H]L-glutamate uptake study.

UNLABELLED: Hypothermia provides neuroprotection that inhibits increases in extracellular glutamate concentration during ischemia; however, the effect of hypothermia on the glutamate transporter is uncertain. A human glial glutamate transporter (hGLT-1) cDNA, isolated by screening a cDNA, library was cloned and stably transfected into Chinese hamster ovary cells. We assessed the effects of temperature on transporter activity in [3H]L-glutamate flux experiments at 23, 32, and 37 degrees C. Hypothermia of 23 degrees C and 32 degrees C decreased [3H]L-glutamate uptake at 60 min, to 76.7%+/-7.3% (P < 0.05, n = 5) and 70.7%+/-7.5% (P < 0.05, n = 5) of uptake at 37 degrees C, respectively. Reversed uptake of preloaded [3H]L-glutamate via hGLT-1 was not observed at any temperature. The specific uptakes (Q10 values) for 37 degrees C to 32 degrees C and 32 degrees C to 23 degrees C at 30 min were 3.48 and 2.37, whereas they were 2.17 and 0.91, respectively, for 60 min. These changes suggest that hypothermia attenuates uptake of extracellular glutamate via hGLT-1 in a temperature- and time-dependent manner. IMPLICATIONS: Under certain pathologic conditions, including cerebral ischemia and traumatic brain injury, glutamate neurotoxicity may initially be propagated by hypothermia due to relative failure of glutamate uptake via Human Glial Glutamate Transporter before a subsequent recovery of uptake.

ATP-Binding Cassette Transporters↗

Occurrence of cello-oligosaccharides in the apoplast of auxin-treated pea stems

Treatment of pea (Pisum sativum L.) hypocotyl segments with indole-3-butyric acid, which promotes segment elongation, increased the solubilization of both xyloglucan and cello-oligosaccharides in the apoplast of auxin-treated pea stems. The cello-oligosaccharides were isolated from the apoplastic solution with a charcoal/Celite column and were identified as cellobiose, cellotriose, and cellotetraose after subsequent thin-layer chromatography and paper electrophoresis. Cello-oligosaccharides in the apoplastic fraction were monitored using cellobiose dehydrogenase. Both xyloglucan and cello-oligosaccharides appeared to be formed concurrently within 30 min after treatment with the auxin, and the cello-oligosaccharides increased with stem elongation even after 2 h. The total activity of cellulase did not increase for up to 4 h.

Journal Article↗

[Plain X-ray diagnosis of noncardiogenic edema].

Noncardiogenic edema is pulmonary edema without increased precapillary blood pressure of the lung. The causes of noncardiogenic edema include decreased blood osmotic pressure, increased permeability of basement membrane, and disturbed lymphatic drainage. We describe the plain X-ray findings of noncardiogenic pulmonary edema as they correlate with the pathophysiology of pulmonary water influx from blood vessels into the extravascular space. We discuss certain selected causes of noncardiogenic edema such as ARDS and high altitude, focusing on the differential diagnosis between cardiogenic edema and noncardiogenic edema and the causes of noncardiogenic edema.

Cell Membrane Permeability↗

Control of expression by the cellulose synthase (bcsA) promoter region from Acetobacter xylinum BPR 2001.

The 5' upstream region (about 3.1kb) of the cellulose synthase operon (bcs operon) has been isolated by cloning from Acetobacter xylinum strain BPR 2001. The expression level of the upstream region was determined using sucrose synthase cDNA as a reporter gene in the shuttle vector pSA19. The expression occurred with the 1.1-kb upstream sequence from the ATG start codon of the bcs operon but not with the 241-bp upstream sequence in A. xylinum, although neither the 1.1-kb nor the 241-bp upstream sequence caused any expression as a promoter in Escherichia coli. The level of expression with the 1. 1-kb upstream sequence in A. aceti was 75% of that in A. xylinum. These results suggest that the upstream region functions as a specific promoter for the Acetobacter genus. The expression was reduced by the introduction of the 241-bp upstream region between the lac promoter and the reporter gene in E. coli and was not detected in A. xylinum. This suggests that the short upstream region composed of 241bp contains the site(s) which causes a negative regulation on the transcription for bcs operon. The production of recombinant protein with the ribosome-binding site (RBS) of A. xylinum obtained from the bcs operon, was reduced to about half in E. coli, and that with the site of the lac promoter was also reduced to about half in A. xylinum. This shows that a species-specific predominance occurs during interaction between mRNA and 16S rRNA in the RBS between A. xylinum and E. coli.

Amino Acid Sequence↗

Intrapulmonary metastasis of lung cancer: soft x-ray investigation of inflated and fixed lung.

BACKGROUND: Intrapulmonary metastasis (IPM) of lung cancer is thought to be an important factor influencing patient prognosis. It is not easy to detect a small IPM by preoperative examination and sometimes even by postoperative pathologic investigation. We applied soft x-ray investigation to inflated and fixed lungs for the detection of IPM. STUDY DESIGN: From 1990 to 1992, 75 patients with lung cancer who had no metastatic lesions on preoperative whole CT, MRI, and technetium-99m bone scintigram examinations underwent lung resection. The resected lungs were fixed in an inflated condition, sliced at the corresponding CT levels into 10-mm-thick sections, and submitted for soft x-ray examination. When an accessory nodular shadow(s) was detected on the soft x-ray images, the size of the nodule and its distance from the primary tumor were measured. RESULTS: In 23 of the 75 patients, accessory nodular shadows were detected on the soft x-ray images. Six nodules in 6 patients proved to be IPM, 2 of which were also detected by postoperative macroscopic examination. Another 2 microscopic IPM were found only by postoperative pathologic examination. The total detection rate of IPM was 10.7% (8 of 75 patients) in this series. The detection rate of IPM at our institute was 5.4% before this study (1979 to 1989). The mean diameter of the IPM detected by the soft x-ray method was 2.8 +/- 1.5 mm, and this was significantly smaller than that of the macroscopically detected nodules (7.2 +/- 3.2 mm). CONCLUSIONS: Our data show that soft x-ray investigation is an effective procedure to detect relatively small intrapulmonary metastatic nodules and will contribute to precise postoperative staging of patients with lung cancer.

Adenocarcinoma↗

Measurement of muscle hardness: a methodological study.

The aim of the present study was to determine the intra- and interobserver variation during measurement of muscle hardness with a new instrument, a so-called hardness meter. In addition, we investigated the factors which may influence the recording of muscle hardness and whether the hardness differs within the same muscle. Hardness was measured at a standardized point on the trapezius muscle in 20 volunteers by 2 observers in random order and again by 1 of the observers after 30 min. Muscle hardness was then measured at 3 different locations on the trapezius muscle. In addition, the muscle hardness at the standard point was measured in 10 volunteers by both observers at 3 different speeds of the applied pressure. The intraobserver variation was 10% and the interobserver variation was 12% for recording of muscle hardness. There was no significant difference in hardness values from time to time within the same observer (p=0.12), while there was a significant difference in hardness measurements between observers (p=0.007). Muscle hardness differed significantly among the 3 anatomical locations (p=0.03). For both observers, there was a significant difference among hardness recordings obtained at 3 different speeds (p=0.01 and p=0.0001). In conclusion, we have shown that the hardness meter can measure muscle hardness reliably if the same observer is used throughout a study. The speed factor was a major source of variability In addition, we have demonstrated that muscle hardness differs within the same muscle. We suggest that the muscle hardness meter will be an important tool in future research of the mechanisms leading to myofascial pain.

Adult↗

An increase in apparent affinity for sucrose of mung bean sucrose synthase is caused by in vitro phosphorylation or directed mutagenesis of Ser11.

A mutational analysis of mung bean (Vigna radiata Wilczek) sucrose synthase was performed by site-directed mutagenesis of the recombinant protein expressed in Escherichia coli, in which two different acidic amino acid residues (Asp or Glu) were introduced at Ser11 (S11D, S11E). Only the wild-type enzyme (Ser11) was phosphorylated in vitro by a Ca(2+)-dependent protein kinase from soybean root nodules, suggesting that this is the specific target residue in mung bean sucrose synthase. The apparent affinity for sucrose was increased in this phosphorylated enzyme and also in the S11D and S11E mutant enzymes, although the affinities for UDP-glucose and fructose were similar in the wild-type, phosphorylated wild-type, and mutant enzymes. These results suggest that a monoanionic (1-) side chain at position 11 mimics the Ser11-P2- residue to bind and cleave sucrose for the synthesis of UDP-glucose. Since the S11E mutant enzyme showed the lowest K(m) (sucrose) and the highest catalytic efficiency of the recombinant proteins, the enzymic properties of this S11E mutant were further characterized. The results showed that replacement of Ser11 with Glu11 modestly protected the sucrose synthesis activity against phenolic glycosides and altered the enzyme nucleotide specificity. We postulate that the introduction of negative charge at Ser11 is possibly involved in the enzymatic perturbation of sucrose synthase.

Amino Acid Sequence↗

[A trial to reduce cardiac motion artifact on HR-CT images of the lung with the use of subsecond scan and special cine reconstruction algorithm].

We describe our trial to reduce cardiac motion artifacts on HR-CT images caused by cardiac pulsation by combining use of subsecond CT (scan time 0.8 s) and a special cine reconstruction algorithm (cine reconstruction algorithm with 180-degree helical interpolation). Eleven to 51 HR-CT images were reconstructed with the special cine reconstruction algorithm at the pitch of 0.1 (0.08 s) from the data obtained by two to six contagious rotation scans at the same level. Images with the fewest cardiac motion artifacts were selected for evaluation. These images were compared with those reconstructed with a conventional cine reconstruction algorithm and step-by-step scan. In spite of its increased radiation exposure, technical complexity and slight degradation of spatial resolution, our method was useful in reducing cardiac motion artifacts on HR-CT images in regions adjacent to the heart.

Algorithms↗

[A trial to reduce cardiac motion artifacts on HR-CT images of lung with combined use of subsecond scan and a high temporal resolution reconstruction algorithm].

PURPOSE: To reduce cardiac motion artifacts with combined use of subsecond CT scanning and a high temporal resolution algorithm. SUBJECTS AND METHODS: Scan data were obtained with step-by-step scanning (scan time, 0.8 seconds). Two sets of images were reconstructed, with and without high temporal resolution reconstruction algorithm (HTRR), from the same set of scan data. HTRR, derived from modification of 180-degree linear helical interpolation, reduces full width at half maximum (FWHM) to half of the scan time. Motion artifacts of two sets of images were compared. RESULTS: The use of HTRR seemed to be useful in reducing cardiac motion artifacts owing to the improvement of effective time resolution. However, there were some images without significant reduction of cardiac motion artifacts. CONCLUSION: Our technique is useful in reducing cardiac motion artifacts without an increase in radiation exposure.

Algorithms↗

FR167653, a dual inhibitor of interleukin-1 and tumor necrosis factor-alpha, ameliorates endotoxin-induced shock.

Increased production of interleukin-1 and tumor necrosis factor-alpha (TNF-alpha) have been implicated in the pathophysiology of a variety of diseases including circulatory shock. The present study evaluated the efficacy of FR167653 (1-[7-(4-fluorophenyl)-1,2,3,4-tetrahydro-8-pyridylpyrazolo[5,1-c] [1,2,4]triazin-2-yl]-2-phenylethanedione sulfate monohydrate), a dual inhibitor of interleukin-1 and TNF-alpha production, to protect rabbits from the shock and lethality induced by lipopolysaccharide. In this sepsis model, FR167653 at a dose of 0.32 mg/kg per h ameliorated the 7-day mortality from 93% in the placebo group to 47% in the FR167653-treated group and, at doses of 0.10-0.32 mg/kg per h, attenuated the hypotensive response to lipopolysaccharide challenge and returned mean arterial blood pressure to almost normal levels. The increases in plasma interleukin-1 and TNF-alpha levels evoked by lipopolysaccharide administration were also inhibited by treatment with FR167653, which was efficacious at doses of 0.1-0.32 mg/kg per h. In addition, FR167653 treatment attenuated the increases in plasma creatinine concentrations consistent with renal damage in a dose-dependent manner. These findings suggested that FR 167653 has a beneficial potential as a drug for septic shock or multiple organ dysfunction syndrome.

Animals↗

Prevalence of migraine in Japan: a nationwide survey.

This study presents the first nationwide survey of migraine in Japan. A representative sample of 4029 subjects aged 15 years or older was selected from the Japanese population according to the quota method. A combination of telephone interview and mailed questionnaire methods was used. Diagnosis of migraine was based on the International Headache Society (IHS) Classification. The overall prevalence of migraine in the past year was 8.4%; 5.8% was migraine without aura and 2.6% was migraine with aura. Significant correlation was found between the prevalence of migraine and such variables as gender, age and district of residence. Doctor attendance rate was very low and 69.4% with migraine had never consulted a physician for headache. Yet, 74.2% complained that migraine headache impaired their daily activity significantly. Only 11.6% were aware that their headache was migraine and 56.9% were using only the over-the-counter drugs. The study revealed a comparably high prevalence of migraine in the general population of Japan compared with other countries. A genetic factor was speculated as the cause of regional difference in migraine prevalence.

Adolescent↗

Changes in levels of mRNAs for cell wall-related enzymes in growing cotton fiber cells.

mRNAs for cell wall-related enzymes in developing cotton fiber cells were measured by reverse transcription-PCR analysis. Both endo-1,4-beta-glucanase and expansin mRNA levels were high during cell elongation but decreased when cell elongation ceased, and xyloglucan decreased. The endo-1,3-beta-glucanase mRNA level was very low in the elongating cells but increased gradually at the onset of secondary wall synthesis, accompanying the massive deposition of cellulose. Endoxyloglucan transferase and sucrose synthase mRNA levels were constant during all stages of growth.

Cell Wall↗

Factors affecting the radiologic appearance of peripheral bronchogenic carcinomas.

The use of high-resolution computed tomography and magnetic resonance imaging have allowed the detailed description of morphologic findings associated with lung cancer. In particular, the desmoplastic response of lung tissue to tumor growth has not been adequately described. This article reviews roentgenologic and pathologic correlations of primary lung carcinomas. An irregular or indistinct tumor margin may be caused by tumor infiltration, an irregular desmoplastic response to the tumor growth, or irregular contraction in the central portion of the tumor. Solid tumor growth, on the other hand, may be associated with a well-defined tumor margin, with or without displacement of adjacent anatomical structures.

Adenocarcinoma↗

Laser isolation of plant sex chromosomes: studies on the DNA composition of the X and Y sex chromosomes of Silene latifolia.

X and Y sex chromosomes from the dioecious plant Silene latifolia (white campion) were isolated from mitotic metaphase chromosome preparations on polyester membranes. Autosomes were ablated using an argon ion laser microbeam and isolated sex chromosomes were then recovered on excised fragments of polyester membrane. Sex chromosome associated DNA sequences were amplified using the degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR) and pools of DOP-PCR products were used to investigate the genomic organization of the S. latifolia sex chromosomes. The chromosomal locations of cloned sex chromosome repeat sequences were analysed by fluorescence in situ hybridization and data complementary to laser ablation studies were obtained by genomic in situ hybridization. In combination, these studies demonstrate that the X and Y sex chromosomes of S. latifolia are of very similar DNA composition and also that they share a significant repetitive DNA content with the autosomes. The evolution of sex chromosomes in Silene is discussed and compared with that in another dioecious species, Rumex acetosa.

DNA, Plant↗

The relations between expiratory chest CT using helical CT and pulmonary function tests in emphysema.

Chest computed tomography (CT) and pulmonary function tests are usually used for the evaluation of emphysema. We performed the present study to evaluate more accurately the relationship between the degree of morphological change and the functional abnormality in emphysema. We used a helical CT apparatus to obtain the CT images both at full inspiration and full expiration. We classed the degree of emphysema into 5 grades (0-4) by visual observation on CT image obtained at the two levels, upper and lower fields, on each side. We assigned each of these grades a visual score. We also calculated the average ratio of CT numbers of each of the lung fields at full expiration to that at full inspiration for each level. We designated this the E/I ratio. We then examined the relationship between the findings acquired by each of these two methods and pulmonary function test results. The visual score showed correlations with FEV1(% predicted) (-0.592, p < 0.0001), RV/TLC(%)(0.425, p < 0.025), and DL(CO)/VA(% predicted)(-0.538, p < 0.0001). The E/I ratio showed correlations with FEV1(% predicted) (-0.620, p < 0.0001), RV/TLC(%)(0.661, p < 0.0001), and DL(CO)/VA(% predicted) (-0.299, p < 0.05). Compared with the E/I ratio, the visual score showed much higher correlation with DL(CO)/VA(% predicted), but the E/I ratio showed stronger correlation with RV/TLC(%) than did the visual score. The visual score may reflect the degree of emphysema, whereas the E/I ratio reflects the air trapping. These findings suggest that the helical CT is useful in the evaluation of emphysematous changes and that the E/I ratio reflects the air trapping.

Aged↗