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J Sasaki

Publications and source records attributed to J Sasaki.

At least 163 records · Page 9Linked to original sources

Differences in mechanisms for cell-free NADPH oxidase activation between arachidonate and sodium dodecyl sulfate.

Mechanisms for the cell-free activation of NADPH oxidase by sodium dodecyl sulfate (SDS) and arachidonate were compared in relation to their responsiveness to short chain diacylglycerols. The plasma membrane and cytosol prepared from guinea pig neutrophils were used for the cell-free system. The activation of NADPH oxidase by SDS was enhanced about 5- to 10-fold by 1,2-dioctanoylglycerol (diC8), but not by either 1,2-dihexanoylglycerol (diC6) or 1,2-didecanoylglycerol (diC10). However, none of these diacylglycerols potentiated the NADPH oxidase activation by arachidonate. The maximal extent of activation by the combination of SDS and diC8 was similar to that by arachidonate alone. In the presence of sufficient amounts of diC8 and SDS, GTP gamma S potentiated the activation of NADPH oxidase. The potentiating activity of diC8 was preserved in the membrane fraction, not in the cytosol fraction. These results suggest that arachidonate may possess the functions of both SDS and diC8 in the activation. In addition, diC8 and GTP gamma S seem to independently enhance the NADPH oxidase activation.

Animals↗

Identification of two apolipoprotein variants, A-I Kaho (Asp 51-->Val) and A-I Lys 107 deletion.

We have identified two apolipoprotein (apo) A-I variants using isoelectric focusing gel electrophoresis: apo A-I Kaho which has a relative charge of +1 compared to normal apo A-I4, and apo A-I Nanakuma2 which has a relative charge of -1. Sequence analysis of PCR-amplified DNA from the proband of apo A-I Kaho revealed a single substitution of aspartic acid (GAC) for valine (GTC) at residue 51. Sequence analysis of PCR-amplified DNA from the proband of apo A-I Nanakuma2 revealed a three-base (AAG or AGA) deletion between bases 186 and 193 from the 5' end of exon 4 that leads to deletion of Lys 106 or 107. This mutation may be the same as that of apo A-I Marburg or A-I Munster-2 reported by Rall et al. (Rall SC Jr, Weisgraber KH, Mahley RW, Ogawa Y, Fielding CJ, Utermann G, Haas J, Steinmetz A, Menzel HJ, and Assmann G, J Biol Chem, 259: 10063-10070, 1984). Because of its unique sequence between 185 and 193 from the 5' end of exon 4 of apo A-I gene, we could not define whether AAG or AGA is deleted by DNA sequencing.

Adult↗

Microbial glycosylation of macrolide antibiotics by Streptomyces hygroscopicus ATCC 31080 and distribution of a macrolide glycosyl transferase in several Streptomyces strains.

In the course of our microbial transformation study on erythromycin derivatives, Streptomyces hygroscopicus ATCC 31080, which produces a polyether antibiotic carriomycin, was found to transform erythromycin derivatives to their inactivated derivatives. The structures of inactivated derivatives prepared by enzyme reaction using the cell extract, UDP-glucose (or UDP-galactose) and Mg2+ (or Mn2+) were elucidated on the basis of analysis of thei spectral data to be the compounds glycosylated at C-2' of a desosamine moiety, indicating that the enzyme is a macrolide glycosyl transferase (MGT). The MGT activity of cell extract from S. antibioticus ATCC 11891, a producing organism of oleandomycin, could be distinguished from that of ATCC 31080, based on the ability to glycosylate tylosin. We examined 32 actinomycete strains producing such polyketides as macrolide and polyether antibiotics, and found that 15 strains of Streptomyces have macrolide glycosyl transferase activity. It suggests that the MGTs have been distributed among at least polyketide producing Streptomyces strains.

Anti-Bacterial Agents↗

[Quality control of surgery in multicenter study--interinstitutional and individual differences].

UNLABELLED: In a prospective randomized controlled trial (PRCT) of adjuvant chemotherapy for gastric cancer patients participated by 6 cancer center hospitals, inter-institutional and inter-individual differences about several factors related to the surgery and remote survivals were analyzed. From 1988 to 1992, 1,049 evaluable patients were enrolled and stratified to 6 groups according to the grade of the serosal invasion and curability, and analyzed statistically. RESULTS: 1) There were no inter-institutional differences about postoperative survival rate, but there were slight differences about patient's background factors and entry policy of each institution. 2) There were no inter-institutional differences about several factors related to the surgery such as operative time, bleeding volume during operation, and the length of the administration periods. 3) There were no inter-individual differences about the length of the administration periods, but there were a little differences about operative time and the volume of hemorrhage. When the PRCT of adjuvant chemotherapy of which the end point is survival would be designed, no inter-institutional differences about remote survival and surgical operation in each institution should be confirmed before starting the trial.

Chemotherapy, Adjuvant↗

Relation of C4b-binding protein to athero-sclerosis of the descending thoracic aorta.

C4b binding protein (C4bp) is a regulator of the classical pathway of the complementing system. It forms a complex with protein S which serves as a cofactor of coagulation inhibitor, protein C. We have reported that C4bp is an acute phase reactant and associated with total cholesterol and triglyceride concentrations (Biochim. Biophys. Acta 963 (1988) 98-108). This suggests a possible association of C4bp with athero-sclerosis. We examined the relation of C4bp levels and the severity of atherosclerosis of the descending thoracic aorta in 98 Japanese men. The severity of aortic atherosclerosis was assessed by average sclerotic length (ASL) and average sclerotic area (ASA), using transesophageal echocardiography. After adjustment for age, C4bp levels increased significantly with increasing ASL and ASA. The association remained significant even after adjusting for total cholesterol, hypertension, smoking, drinking, body mass index, fasting blood sugar, and uric acid. Immunohistochemical analysis of specimens of the descending thoracic aorta from autopsies, demonstrated the presence of C4bp in the foamy macrophages of fatty streaks and the necrotic core of atheromatous plaque. These findings indicate that the serum level of C4bp can serve as an independent indicator of aortic athero-sclerosis.

Adult↗

[Thoracoscopic closure of patent ductus arteriosus for children: two case report].

We reported thoracoscopic closure of patent ductus arteriosus (TCPDA) performed on two children. All of two patients had no residual shunt postoperatively. Postoperative pain was less than left thoracotomy procedure. In addition TCPDA had merits cosmetically. We concluded that TCPDA was less invasive and safety procedure compared with conventional procedure.

Child, Preschool↗

[A case of pulmonary sarcoma with a differential diagnosis from preoperative pulmonary thromboembolism].

Primary sarcoma of pulmonary artery is a rare tumor. A 46-year-old man was hospitalized because of dyspnea and hemoptysis. Pulmonary thromboembolism was suspected by chest X-ray, scintigraphy, and CT. But the mass had a to-and-fro movement above the pulmonary valve on pulmonary arterial cineangiography. We strongly suspected that it was primary neoplasm of pulmonary artery. The pulmonary artery was opened under cardiopulmonary bypass. The tumor arised pulmonary trunk and extended peripherally. The tumor was surgically removed as complete as possible, though a proxymal balloon embolectomy was unsuccessful. The histological finding was malignant fibrous histiocytoma of the pulmonary artery.

Cardiopulmonary Bypass↗

[A case of poorly differentiated ovarian serous adenocarcinoma responding favorably to etoposide and cisplatin].

A case of primary ovarian cancer responding favorably to EP therapy is reported. The patient is a 41-year-old woman with stage IV poorly differentiated serous papillary adenocarcinoma of the ovary. At 21 days after the optimal initial surgical exploration, multiple tumor nodules recurred in peritoneal cavity, and one of them had a diameter of 5 cm. Since we had found previously that poorly differentiated adenocarcinoma of the ovary responded to etoposide by a disease-oriented screening system, the patient was treated by combination chemotherapy (etoposide 100 mg/body/day x 5 days + cisplatin 20 mg/body/day x 5 days) as first line chemotherapy. After 2 cycles of chemotherapy, the tumor disappeared on CT, and the serum CA 125 level decreased to 14 U/ml from 2,050 U/ml. She was evaluated as a complete response after 5 cycles of combination chemotherapy, and only mild side-effects were recognized. These results demonstrate that the combination chemotherapy of etoposide and cisplatin might be recommended as first-line chemotherapy for poorly differentiated serous adenocarcinoma of the ovary.

Adult↗

Endoscopic screening for esophageal cancer in 788 patients with head and neck cancers.

Esophageal cancer has a poor prognosis because it is difficult to detect in its early stages and, even if an operation is possible, the postoperative quality of life is much impaired. An early diagnosis can lead to a good prognosis and enables treatment by endoscopic mucosal resection (EEMR), contributing to a postoperative good quality of life. As head and neck cancers are known to have a high risk of concomitant esophageal cancer, endoscopic screening with iodine staining was performed on 788 patients with head and neck cancers. Among them, 93 cases of esophageal cancers (11.8%) and 23 cases of gastric cancers (2.9%) were detected. Seventy-two cases (77.4%) of the 93 esophageal cancers were superficial cancers limited to the submucosal layer. Twenty cases, treated by EEMR, had a good postoperative course without local recurrence. We suggest that endoscopic screening for esophageal cancer should be performed on all patients with head and neck cancers, because it allows early detection and a good prognosis, and the treatment can be completed by endoscopic maneuver.

Aged↗

Structural changes in the lumirhodopsin-to-metarhodopsin I conversion of air-dried bovine rhodopsin.

Structural changes during the photochemical reactions of unhydrated air-dried films of bovine rhodopsin in rod outer segments were examined by visible and Fourier transform infrared (FTIR) spectroscopy at 200, 240, and 280 K. These films exhibited conversion from a lumirhodopsin state to a metarhodopsin I state with a time constant of 13.5 min at 280 K, but did not form metarhodopsin II at all, as observed earlier for digitonin-extracted rhodopsin in dry gelatin films [Wald, Durell, and St. George (1950) Science 111, 179-181]. Lumirhodopsin which was stable in the dry film was very similar to normal lumirhodopsin. The metarhodopsin I-like state retained properties characteristic of lumirhodopsin in regard to a twisted structure between the C14-H and the Schiff base of the chromophore, and perturbation around Glu122, although the C-C stretch frequencies of the chromophore were identical with those of metarhodopsin I. Thus, under dry conditions some of the structural changes that lead to metarhodopsin I are partially inhibited. These defects could result in stable lumirhodopsin and the failure to form metarhodopsin II, which is in equilibrium with metarhodopsin I.

Animals↗

Glutamic acid 204 is the terminal proton release group at the extracellular surface of bacteriorhodopsin.

We have measured proton release into the medium after proton transfer from the retinal Schiff base to Asp85 in the photocycle and the C = O stretch bands of carboxylic acids in wild type bacteriorhodopsin and the E204Q and E204D mutants. In E204Q, but not in E204D, the normal proton release is absent. Consistent with this, a negative band in the Fourier transform infrared difference spectra at 1700 cm-1 in the wild type, which we now attribute to depletion of the protonated E204, is also absent in E204Q. In E204D, this band is shifted to 1714 cm-1, as expected from the higher frequency for a protonated aspartic than for a glutamic acid. Consistent with their origin from protonated carboxyls, the depletion bands in the wild type and E204D shift in D2O to 1690 and 1703 cm-1, respectively. In the protein structure, Glu204 seems to be connected to the Schiff base region by a chain of hydrogen-bonded water. As with other residues closer to the Schiff base, replacement of Glu204 with glutamine changes the O-H stretch frequency of the bound water molecule near Asp85 that undergoes hydrogen-bonding change in the photocycle. The results therefore identify Glu204 as XH, the earlier postulated residue that is the source of the released proton during the transport, and suggest that its deprotonation is triggered by the protonation of Asp85 through a network that contains water dipoles.

Aspartic Acid↗

Light-driven chloride ion transport by halorhodopsin from Natronobacterium pharaonis. 1. The photochemical cycle.

The photochemical cycle of the light-driven chloride pump, halorhodopsin from N. pharaonis, is described by transient optical multichannel and single-wavelength spectroscopy in the visible, and in the infrared. Titration of a blue-shift of the absorption maximum upon addition of chloride describes a binding site with a KD of 1 mM. The reaction sequence after the all-trans to 13-cis photoisomerization of the retinal in this chloride binding form is itself dependent on chloride. At 2 M chloride it is described by the scheme: HR-->K<==>L<==>N-->HR that relaxes in a few milliseconds, and is very similar to the photocycle of bacteriorhodopsin under conditions where the retinal Schiff base cannot deprotonate. At lower chloride concentrations, e.g., 0.1 M, however, a red-shifted state termed O appears between N and HR, in equilibrium with N. The absorption spectra of K, L, N, and O are very similar to their counterparts in the bacteriorhodopsin photocycle. As in their equivalents in bacteriorhodopsin, in the N state the retinal is still 13-cis, but it is reisomerized in the O state to all-trans.

Archaea↗

The complex extracellular domain regulates the deprotonation and reprotonation of the retinal Schiff base during the bacteriorhodopsin photocycle.

During the L-->M reaction of the bacteriorhodopsin photocycle the proton of the retinal Schiff base is transferred to the anionic D85. This step, together with the subsequent reprotonation of the Schiff base from D96 in the M-->N reaction, results in the translocation of a proton across the membrane. The first of these critical proton transfers occurs in an extended hydrogen-bonded complex containing two negatively charged residues (D85 and D212), two positively charged groups (the Schiff base and R82), and coordinated water. We simplified this region by replacing D212 and R82 with neutral residues, leaving only the proton donor and acceptor as charged groups. The D212N/R82Q mutant shows essentially normal proton transport, but in the photocycle neither of this protein nor of the D212N/R82Q/D96N triple mutant does a deprotonated Schiff base (the M intermediate) accumulate. Instead, the photocycle contains only the K, L, and N intermediates. Infrared difference spectra of D212N/R82Q and D212N/R82Q/D96N demonstrate that although D96 becomes deprotonated in N, D85 remains unprotonated. On the other hand, M is produced at pH > 8, where according to independent evidence the L<==>M equilibrium should shift toward M. Likewise, M is restored in the photocycle when the retinal is replaced with the 14-fluoro analogue that lowers the pKa of the protonated Schiff base, and now D85 becomes protonated as in the wild type.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteriorhodopsins↗

Conversion of bacteriorhodopsin into a chloride ion pump.

In the light-driven proton pump bacteriorhodopsin, proton transfer from the retinal Schiff base to aspartate-85 is the crucial reaction of the transport cycle. In halorhodopsin, a light-driven chloride ion pump, the equivalent of residue 85 is threonine. When aspartate-85 was replaced with threonine, the mutated bacteriorhodopsin became a chloride ion pump when expressed in Halobacterium salinarium and, like halorhodopsin, actively transported chloride ions in the direction opposite from the proton pump. Chloride was bound to it, as revealed by large shifts of the absorption maximum of the chromophore, and its photointermediates included a red-shifted state in the millisecond time domain, with its amplitude and decay rate dependent on chloride concentration. Bacteriorhodopsin and halorhodopsin thus share a common transport mechanism, and the interaction of residue 85 with the retinal Schiff base determines the ionic specificity.

Aspartic Acid↗

Water structural changes at the proton uptake site (the Thr46-Asp96 domain) in the L intermediate of bacteriorhodopsin.

Fourier transform infrared spectra of the L intermediate of light-adapted bacteriorhodopsin were examined for recombinant proteins with amino acid substitutions at Thr46 and Asp96. Two O-H stretching vibrational bands of water, at 3607 and 3577 cm-1, change into stronger H-bonding states in L of the wild type. Thr46-->Val substitution abolished these bands in spite of the fact that [3-18O]threonine-labeling did not shift them, indicating that they correspond to coordination of the water with Thr46. The two water bands were restored, although with changed frequencies, by an additional Asp96-->Asn substitution in Thr46-->Val/Asp96-->Asn. A single Asp96-->Asn substitution abolished the 3607 cm-1 band. Thus, Asp96 also takes part in structural changes in water. The perturbations of these water molecules in the L intermediate displayed a weak correlation with the ratio of intensity change in the two vibrational bands of the Schiff base mode at 1312 and 1301 cm-1 and the rate for the deprotonation of the Schiff base at the L-to-M reaction of the photocycle. We find, therefore, that the water molecules in the cytoplasmic Asp96-Thr46 domain, which comprises the site of proton uptake after formation of the M intermediate, undergo structural changes in the L intermediate already. These changes are transmitted to the extracellular domain and affect interaction of the Schiff base with Asp85, that is far removed from this region.

Aspartic Acid↗

Effects of surface modification of materials on human neutrophil activation.

The properties of a polymer surface affect the cellular functions and morphology of cells in contact with the polymer. In this paper, we will demonstrate the effects of surface modification of materials on various neutrophil markers of activation. The sulfonation of a polystyrene surface caused increases in its negative charge and hydrophilicity. The sulfonation did not affect the number of adhered neutrophils, but the shape of the neutrophils adhered on the material was different; a round shape on highly sulfonated polystyrene and a spread shape on weakly sulfonated or non-sulfonated polystyrene. Expression of the adhesion molecule, CD11b, on neutrophils was also affected by the properties of the polymer surface. CD11b was expressed in neutrophils adhered on polystyrene and the expression decreased with increasing sulfonation of the surface. The expression of CD11b on the neutrophils on highly sulfonated polystyrene was the same as that on non-adhered neutrophils. In contrast, the expression of CD11a was not affected by the properties of the material surface. The F-actin content of activated neutrophils and the production of active oxygen groups detected by means of luminol-dependent chemiluminescence were also dependent on the sulfo-group content of the material surface. Finally, the translocation of protein kinase C (PKC) was determined in neutrophils adhered to these materials. Compared to non-adhered cells, the ratio of membrane bound to cytosolic PKC increased in adhered cells, but the increase was suppressed by sulfonation of the material surface. These data suggest that activation of neutrophils on polystyrene is suppressed by surface modification with increasing negative charge and/or hydrophilicity.

Actins↗

Interaction of tryptophan-182 with the retinal 9-methyl group in the L intermediate of bacteriorhodopsin.

An intense indole N-H stretching vibrational band at 3486 cm-1 in the difference Fourier transform infrared spectrum is one of the characteristic features of the L intermediate of bacteriorhodopsin [Maeda, Sasaki, Ohkita, Simpson, & Herzfeld (1992) Biochemistry 31, 12543]. This band is now assigned to tryptophan-182. The Trp182-->Phe (W182F) protein shows specific features in the difference spectrum in the visible region upon L formation, and exhibits great delay in the L-M conversion. Fourier transform infrared difference spectra further indicate that while the intensity of the C-methyl in-plane bending vibration at 1009 cm-1 is lost in the L intermediate of the wild type, its intensity remains high in the W182F protein. The intensity of the N-H stretching vibration upon L formation is diminished considerably in an artificial bacteriorhodopsin containing 9-desmethylretinal. It also exhibits delayed M formation. These results suggest that Trp182 interacts with the retinal side chain through the 9-methyl group, and thereby affects the L-to-M conversion.

Bacteriorhodopsins↗