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Biomedical subjects

Y Miyagawa

Publications and source records attributed to Y Miyagawa.

At least 19 recordsLinked to original sources

Differential expression of succinyl CoA transferase (SCOT) genes in somatic and germline cells of the mouse testis.

Succinyl CoA:3-oxo acid CoA transferase (SCOT/OXCT; EC 2.8.3.5) is a key mitochondrial enzyme in the metabolism of ketone bodies in various organs (but not in the liver). We identified a cDNA clone of the testicular germ cell-specific succinyl CoA transferase isozyme (SCOT-t). We then isolated a mouse orthologue of the SCOT/OXCT cDNA (SCOT-s) and determined the expression of the two types of SCOT in the testis. The mRNAs of scot-s and scot-t were expressed exclusively in testicular somatic cells (i.e. Leydig and Sertoli cells) and germ cells, respectively. SCOT enzymatic activities were assayed in Leydig cell (SCOT-s) and sperm (SCOT-t) fractions. The SCOT activity in sperm was 2.5-fold higher than that in Leydig cells. We conclude that germ cells and somatic cells differentially express the SCOT enzymes and that the SCOT activity of sperm caused exclusively by SCOT-t should play an important role in sperm activity.

Animals↗

Conventional multiple or microdissection testicular sperm extraction: a comparative study.

BACKGROUND: Testicular sperm extraction (TESE) with ICSI is becoming the first-line treatment for non-obstructive azoospermia (NOA). Recently, the sperm retrieval rate (SRR) by microdissection TESE was reported to be higher than by conventional TESE. However, a comprehensive comparison between multiple and microdissection TESE patients including histological findings has not been reported. METHODS: Patients with NOA who underwent microdissection TESE (n = 56) or multiple TESE (n = 37) were compared. Pre-operative characteristics were similar between groups. In addition, microscopic findings during microdissection TESE also were investigated. RESULTS: Operative time was significantly longer for microdissection TESE than for multiple TESE. Histological examination suggested that spermatogenesis was relatively more impaired in the microdissection TESE group than in the multiple TESE group. Despite this, SRR by microdissection TESE (42.9%) appeared higher than by conventional TESE (35.1%) although this observation failed to reach statistical significance. Seventeen of 26 patients (65.4%) with heterogeneous tubule were successful for sperm retrieval. No severe operative complications occurred in any patient in either group, and no patient required post-operative hormone replacement to treat hypogonadism. CONCLUSIONS: Microsurgical technique is safe and may improve SRR for TESE in a variety of patients with NOA, especially patients with heterogeneous testicular tubules.

Adult↗

Cloning and rational mutagenesis of kexstatin I, a potent proteinaceous inhibitor of Kex2 proteinase.

Kexstatin I is a potent proteinaceous inhibitor of Kex2 proteinase (EC 3.4.21.61). In the present study we show the molecular cloning, primary structure determination and expression of the gene encoding kexstatin I. We also demonstrate its enhanced activity and specificity for Kex2 proteinase inhibition by rational mutagenesis. The cloned kexstatin I gene encoded a protein of 145 amino acid residues, including the 35-residue signal sequence for secretion. The amino acid sequence showed 52% identity with those of the Streptomyces subtilisin inhibitors (SSIs). Thus kexstatin I is the first SSI-family member that can inhibit Kex2 proteinase. The reactive site of the inhibitor was determined to be -Thr(69)-Lys(70) downward arrowGlu(71)-, where downward arrow indicates the reactive site. Because Kex2 proteinase generally shows the highest affinity for substrates with basic amino acid residues at the P(1) and P(2) sites, conversion of the Thr(69)-Lys(70) segment of the inhibitor into dibasic motifs was expected to result in enhanced inhibitory activities. Thus we constructed kexstatin I mutants, in which the Thr(69)-Lys(70) sequence was replaced by the Thr(69)-Arg(70), Lys(69)-Lys(70) and Lys(69)-Arg(70) sequences using PCR-based mutagenesis, and analysed them kinetically. Among these mutants, the Lys(69)-Arg(70) mutant was the most potent inhibitor. The K(i) for Kex2 proteinase was 3.2x10(-10) M, which was 140-fold lower than that of the inhibitor with the Thr(69)-Lys(70) sequence. Although kexstatin I could also inhibit subtilisin, the enhancement of inhibitory activity upon such mutations was specific for Kex2 proteinase inhibition.

Amino Acid Sequence↗

Overexpression of a cyanobacterial fructose-1,6-/sedoheptulose-1,7-bisphosphatase in tobacco enhances photosynthesis and growth.

Transgenic tobacco plants expressing a cyanobacterial fructose-1,6/sedoheptulose-1,7-bisphosphatase targeted to chloroplasts show enhanced photosynthetic efficiency and growth characteristics under atmospheric conditions (360 p.p.m. CO2). Compared with wild-type tobacco, final dry matter and photosynthetic CO2 fixation of the transgenic plants were 1.5-fold and 1.24-fold higher, respectively. Transgenic tobacco also showed a 1.2-fold increase in initial activity of ribulose 1,5 bisphosphate carboxylase/oxygenase (Rubisco) compared with wild-type plants. Levels of intermediates in the Calvin cycle and the accumulation of carbohydrates were also higher than those in wild-type plants. This is the first report in which expression of a single plastid-targeted enzyme has been shown to improve carbon fixation and growth in transgenic plants.

Carbon↗

Bactericidal/permeability-increasing protein promotes complement activation for neutrophil-mediated phagocytosis on bacterial surface.

The neutrophil bactericidal/permeability-increasing protein (BPI) has both bactericidal and lipopolysaccharide-neutralizing activities. The present study suggests that BPI also plays an important role in phagocytosis of Escherichia coli by neutrophils through promotion of complement activation on the bacterial surface. Flow cytometric analysis indicated that fluorescein-labelled E. coli treated with BPI were phagocytosed in the presence of serum at two- to five-fold higher levels than phagocytosis of the bacteria without the treatment. In contrast, phagocytosis of the fluoresceined bacteria with or without treatment by BPI did not occur at all in the absence of serum. The phagocytosis stimulated by BPI and serum was dose-dependent. The effect of BPI on phagocytosis in the presence of serum was not observed on Gram-positive bacteria (Staphylococcus aureus). Interestingly, the complement C3b/iC3b fragments were deposited onto the bacterial surface also as a function of the BPI concentration under conditions similar to those for phagocytosis. Furthermore, the BPI-promoted phagocytosis was blocked completely by anti-C3 F(ab')(2) and partially by anti-complement receptor (CR) type 1 and/or anti-CR type 3. These findings suggest that BPI accelerates complement activation to opsonize bacteria with complement-derived fragments, leading to stimulation of phagocytosis by neutrophils via CR(s).

Antimicrobial Cationic Peptides↗

Renal artery aneurysm causing hydronephrosis.

A 42-year-old man presented with left hydronephrosis incidentally discovered on abdominal echogram during a routine health examination. Color Doppler ultrasonography, intravenous pyelography and angiography revealed a non-calcified renal artery aneurysm of 30 mm in size compressing the pyeroureteral junction and causing hydronephrosis. Three-dimensional computed tomography (3-D CT) using spiral CT clearly displayed the aneurysm located at the first bifurcation of the left renal artery and involving the anterior segmental artery. Decompression was successfully obtained via in situ revascularization of the renal artery after aneurysmectomy. A literature search revealed 12 cases of renal artery aneurysm causing hydronephrosis reported in Japan, although only three cases were documented in other countries, and these reports are reviewed. Use of 3-D CT for evaluation of renal artery aneurysm is advocated.

Adult↗

Testosterone suppresses spermatogenesis in juvenile spermatogonial depletion (jsd ) mice.

Male juvenile spermatogonial depletion (jsd/jsd) mice are sterile because of a failure of spermatogonial differentiation. We have previously reported the recovery of spermatogonial differentiation by suppressing the levels of gonadotropins and testosterone with Nal-Glu, a GnRH antagonist. To determine whether suppression of testosterone or the gonadotropins was responsible for spermatogenic recovery, we examined the effect of supplementation of LH or FSH along with Nal-Glu treatment. Systemic administration of flutamide, an androgen receptor antagonist, was also examined. LH supplementation elevated both serum and intratesticular testosterone levels and suppressed the recovery of spermatogonial differentiation in a dose-dependent manner. Supplementation with FSH did not affect either testosterone levels or spermatogonial differentiation. Furthermore, the mice treated with flutamide showed some recovery of spermatogonial differentiation. The overall findings revealed that testosterone action mediated by androgen receptors suppressed the spermatogonial differentiation in jsd/jsd mice and suggested that spermatogonial differentiation in the jsd mutant is highly sensitive to testosterone suppression.

Androgen Antagonists↗

Development of Ag-Pd-Au-Cu alloys for multiple dental applications. Part 2. Mechanical properties of experimental Ag-Pd-Au-Cu alloys containing Sn or Ga for ceramic-metal restorations.

Eighteen Ag-Pd-Au-Cu alloys, consisting of nine Ag-Pd-Au-Cu mother compositions (Pd: 20, 30 or 40%, Au: 20%, Cu: 10, 15 or 20%, Ag: balance) containing either 5% Sn or 5% Ga as an additive metal, were experimentally prepared. Tensile strength, proof stress, elongation, elastic modulus, and Vickers hardness of these alloys were evaluated to clarify the potential of these alloys for use as ceramic-metal restorations as well as the effects of the Pd and Cu contents on their mechanical properties. The tensile strength, proof stress, elongation, elastic modulus and Vickers hardness of the 18 experimental alloys were in the range of 410.0-984.0 MPa, 289.7-774.3 MPa, 2.2-23.7%, 81.3-123.0 GPa and 135.7-332.3 HV1, respectively. Ten of the 18 experimental alloys can be used for ultra-low fusing ceramics based on their proof stress, elastic modulus, elongation and hardness. Between the Ga- and Sn-added alloys, differences in tensile strength, proof stress, elongation and hardness were found at several Ag-Pd-Au-Cu compositions.

Analysis of Variance↗

Identification of hepatic venous territories in liver resection by using color Doppler ultrasonography: report of two cases.

OBJECTIVE: Resection of the hepatic vein because of the proximity of tumors may result in increased congestion in the noncancerous parenchyma, which in turn may lead to functional hepatic volume loss and postoperative hepatic failure, especially in a case of low hepatic reserve. However, to our knowledge, no technique for estimating the extent of dependent hepatic venous territories before hepatic resection has been established. We examined the possibility of using color Doppler ultrasonography for this purpose. METHODS: A color Doppler system and a linear array transducer equipped with multiple Doppler frequencies ranging from 7 to 13 MHz were used intraoperatively. Two patients with hepatocellular carcinomas were examined. By tracking entire branches of the targeted vessel, from the trunk to the terminal branches extending to the liver surface, it was possible for the boundaries of the dependent areas to be projected and marked on the liver surface with either ink or electrocautery. RESULTS: In both cases, this method was effective for either minimizing the congestive area or preserving the hepatic mass that was being drained via aberrant routes. CONCLUSIONS: Identification of hepatic venous territories by means of color Doppler ultrasonography may provide new information about intrahepatic blood circulation and may increase the safety and curability of hepatic resection.

Aged↗

Evaluation of the defense system in chloroplasts to photooxidative stress caused by paraquat using transgenic tobacco plants expressing catalase from Escherichia coli.

We evaluated the defense system in chloroplasts to photooxidative stress imposed by paraquat treatment under illumination in transgenic tobacco plants with increased tolerance to drought stress at a high light intensity produced by catalase from Escherichia coli targeted to chloroplasts [Shikanai et al. (1998) FEBS Lett. 428: 47]. At 24 h after the paraquat application, Chl was destroyed in the wild-type plants, but not in transgenic plants. Photosynthetic activities monitored by CO2 fixation and Chl fluorescence were much less affected by the paraquat treatment in transgenic lines. The activities of chloroplastic ascorbate peroxidase (APX) isoenzymes decreased in parallel with the depletion of ascorbate (AsA) in leaves in both lines. Paraquat treatment had no effect on the transcript level of chloroplastic APX isoenzymes, while it significantly lowered the level of their proteins. These data suggest that the depletion of AsA in chloroplasts under severe stress conditions inactivates and degrades chloroplastic APX isoenzymes.

Ascorbate Peroxidases↗

Long-term outcome of chronic hepatitis B in adolescents or young adults in follow-up from childhood.

BACKGROUND: It has not yet been defined whether children with chronic hepatitis B are likely to develop severe liver disease in the future. The purpose of this study was to evaluate the evolution of chronic hepatitis B acquired in childhood. METHOD: Fifty-two children in the age range of 0 to 15 years who were positive for hepatitis B surface antigen and hepatitis B e antigen in serum for at least 6 months were enrolled in this study. In the majority of the 52 children, hepatitis B virus infection was acquired by perinatal transmission. All 52 showed abnormal liver function test findings for more than 6 months before enrollment, and the subjects were followed up longitudinally for 3 to 22 years (mean, 11 years). They are now more than 15 years of age (15-27 years old). RESULTS: During the follow-up period, 26 (50%) children had spontaneous seroconversion to anti-hepatitis B e. Serum levels of alanine aminotransferase normalized in these 26 children. In one child of these children, hepatocellular carcinoma developed at the age of 21 years, 16 years after seroconversion, although his liver function profiles remained normal. The other 26 children remained hepatitis B e antigen positive, most with unchanged biochemical features. Sixteen (62%) children among these 26 children were treated with interferon-alpha. Eleven (69%) children had seroconversion to anti-hepatitis B e within the first year after the cessation of therapy. Hepatocellular carcinoma developed in 1 of these 11 children at the age of 16 years, 6 years after interferon therapy. Thus, hepatocellular carcinoma developed in two children in an anti-hepatitis B e positive phase. CONCLUSION: All children carrying hepatitis B surface antigen should be observed carefully to monitor the possible development of hepatocellular carcinoma, especially in the antihepatitis B e-positive phase after spontaneous seroconversion or even after interferon treatment.

Adolescent↗

New initiation system for resin polymerization using metal particles and 4-META.

Fifteen kinds of metal particles were examined to establish whether they could induce the setting of UDMA-based monomer containing BPO without amine under the presence of 4-META at room temperature. FT-IR spectra of the resultant set samples and the monomer were analyzed to see if the setting was caused by the polymerization. The effects of 4-META and BPO concentrations on the setting time were also studied using the metal particles that induced the setting very effectively. As-received Cu, Zn, Mo, Sn, Co, and In particles could initiate the polymerization of the monomer in combination with BPO and 4-META when they were moistened with water. All the three kinds of silver alloy particles examined also could initiate the polymerization, although pure silver metal particles could not. The presence of 4-META drastically shortened the setting time of the mixture of Cu particles and the monomer containing BPO, while higher concentration of BPO in the monomer significantly shortened the setting time.

Acrylic Resins↗

Polymerization of UDMA using zinc particles and 4-META with and without BPO.

The polymerization phenomena of zinc particles moistened with a small amount of water, 4-META, and UDMA without amine in the presence and absence of BPO were investigated. The effects of 4-META and BPO on the setting time and the degree of conversion (DC) were studied. Moreover, the effect of zinc ion amount on the setting time was investigated. As-received zinc particles could induce the polymerization either with or without BPO. A higher concentration of 4-META shortened the setting time and increased DC when BPO was absent. However, the presence of BPO generally retarded the setting time and decreased DC, although its effect was dependent on the 4-META concentration. A higher amount of zinc ion retarded the setting reaction in the presence of 4-META. The zinc particles mixed with 10% zinc sulfate and acetic acid solutions could induce the polymerization of UDMA containing BPO when the amine and 4-META were absent.

Analysis of Variance↗

Development of Ag-Pd-Au-Cu alloy for multiple dental applications. Part 1. Effects of Pd and Cu contents, and addition of Ga or Sn on physical properties and bond with ultra-low fusing ceramic.

Ag-Pd-Au-Cu quaternary alloys consisting of 30-50% Ag, 20-40% Pd, 10-20% Cu and 20% Au (mother alloys) were prepared. Then 5% Sn or 5% Ga was added to the mother alloy compositions, and another two alloy systems (Sn-added alloys and Ga-added alloys) were also prepared. The bond between the prepared alloys and an ultra-low fusing ceramic as well as their physical properties such as the solidus point, liquidus point and the coefficient of thermal expansion were evaluated. The solidus point and liquidus point of the prepared alloys ranged from 802 degrees C to 1142 degrees C and from 931 degrees C to 1223 degrees C, respectively. The coefficient of thermal expansion ranged from 14.6 to 17.1 x 10(-6)/degrees C for the Sn- and Ga-added alloys. In most cases, the Pd and Cu contents significantly influenced the solidus point, liquidus point and coefficient of thermal expansion. All Sn- and Ga-added alloys showed high area fractions of retained ceramic (92.1-100%), while the mother alloy showed relatively low area fractions (82.3%) with a high standard deviation (20.5%). Based on the evaluated properties, six Sn-added alloys and four Ga-added alloys among the prepared alloys were suitable for the application of the tested ultra-low fusing ceramic.

Analysis of Variance↗

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (1998). I. Susceptibility distribution].

The frequencies of bacterial isolation and susceptibilities to antimicrobial agents were investigated on 538 bacterial strains isolated from patients with urinary tract infections (UTIs) in 9 hospitals during the period of June 1998 to May 1999. Of the above bacterial isolates, Gram-positive bacteria accounted for 30.3% and Gram-negative bacteria accounted for 69.7%. Susceptibilities of several isolated bacteria to antimicrobial agents were as follows; against Enterococcus faecalis isolated from patients with UTIs, vancomycin (VCM), ampicillin (ABPC) and imipenem (IPM) had strong activities. Among E. faecalis strains, those with low susceptibilities to all drugs have increased in 1998, compared with those in 1997. VCM showed the highest activity against MRSA isolated from patients with UTIs. The MICs of VCM for all 34 strains were equal to or lower than 2 micrograms/ml. Arbekacin (ABK) was also active against MRSA with the MIC90s of 2 micrograms/ml. Against Escherichia coli and Klebsiella pneumoniae, all drugs except penicillins were active. Particularly, meropenem (MEPM) showed the highest activity with the MICs of 0.125 micrograms/ml or below. Almost all the drugs except minocycline (MINO) showed high activities against Proteus mirabilis. Against Pseudomonas aeruginosa, all drugs were not so active, with the MIC90s of 16 micrograms/ml or above. MEPM, IPM and gentamicin (GM) showed high activities against Serratia marcescens. Generally, it seemed that resistant strains of S. marcescens had decreased since 1996.

Anti-Bacterial Agents↗

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (1998). II. Background of patients].

Clinical background was investigated on 449 patients with urinary tract infections (UTIs) from whom 591 bacterial strains were isolated in 9 hospitals during the period from June, 1998 through May, 1999. About distribution of age and sex of patients and type of infections, among males, patients less than 50 years old were few, and uncomplicated UTIs without indwelling catheters was most frequent. Among females, patients less than 20 years old were few, and uncomplicated was most frequent. Escherichia coli was the most frequently isolated in uncomplicated UTIs, and the higher the ages of patients, the higher were became the isolation frequencies of Enterococcus faecalis, Proteus spp. and Klebsiella spp. In complicated UTIs with indwelling catheters and without indwelling catheters, the types of pathogens had no relation with ages. The complication of infections had decreased E. coli but that had increased Proteus spp., Pseudomonas aeruginosa and Staphylococcus aureus. Until last year, use of antibiotics had decreased pathogens isolated from patients with uncomplicated UTIs drastically in our study. But, pathogens isolated after antibiotics had increased in 1998. As for surgical procedures and types of causative organisms in UTIs, E. faecalis were more isolated when surgical procedures were experienced, and E. coli were more isolated when they were not in uncomplicated and complicated UTIs without indwelling catheters. In complicated UTIs with indwelling catheters, types of causative organisms had no relationship with surgical procedures.

Adult↗

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (1998). III. Secular changes in susceptibility].

Susceptibilities to various antimicrobial agents were examined for Enterococcus faecalis, Staphylococcus aureus, Echerichia coli, Klebsiella spp. and Pseudomonas aeruginosa that were isolated from patients with urinary tract infections (UTIs) in 9 hospitals during June 1998 to May 1999, and the results were compared with those obtained during the same period from 1990 to 1997 in uncomplicated UTIs and complicated UTIs. Among E. faecalis strains, those with low susceptibilities to almost drugs have increased in the latest period. All 5 S. aureus strains isolated from uncomplicated UTIs were the most susceptible to gentamicin (GM). Over 50% of S. aureus strains isolated from complicated UTIs were susceptible to GM, and on the contrary the resistant strains have increased with the MIC90 of 256 micrograms/ml or above. Among S. aureus strains isolated from complicated UTIs, those with low susceptibilities to arbekacin (ABK) have increased in the latest period compared to those during period of 1996-1997, and the MIC90s of them have changed into the lower state from 1 microgram/ml in 1996-1997 to 4 micrograms/ml in 1998. S. aureus strains have continued high susceptibilities to vancomycin (VCM). The susceptibilities to minocycline (MINO) of E. coli showed MIC90: 4 micrograms/ml in 1997, but those have returned in the latest period in uncomplicated UTIs. The MIC90s of ofloxacin (OFLX) to E. coli isolated from uncomplicated and complicated UTIs have been lower 2-3 classes in the latest period than those in 1997. Among Klebsiella spp. strains isolated from uncomplicated UTIs, those with low susceptibilities to almost cephems had increased in 1997, but few of them were detected in the latest study. The sensitive strains of P. aeruginosa to almost drugs have increased during the latest period. The MIC50s of cefozopran (CZOP) and OFLX against P. aeruginosa were the best in our history. The sensitive strains of P. aeruginosa to ceftazidime (CAZ) have increased and its percentage was 30%. Piperacilline (PIPC), cefoperazone (CPZ), GM and OFLX resistant P. aeruginosa strains have increased in the latest period.

Anti-Bacterial Agents↗

[Renocolic fistula: a case report].

A 78-year-old woman had disregarded pneumaturia since April 1998. In March 1999, computed tomography and barium enema were done to examine anemia and positive fecal occult blood, revealing a left renocolic fistula and bilateral renal stones. Intravenous pyelography revealed a left staghorn calculus, non-functioning kidney, and right partial staghorn calculus. Urinalysis showed pyuria and the culture grew Proteus vulgaris and Klebsiella oxytoca. Smear and culture of the urine were negative for acid-fast bacilli. In consideration of the patient's age and conservation of right renal function, right pyelolithotomy was performed first. Three weeks later, left nephrectomy and partial colectomy were done. The postoperative course was uneventful. A renocolic fistula is relatively rare and to our knowledge there have been 37 cases reported in Japan, including our case. Surgery is the main therapy and was performed in 31 patients. Among them, surgery was not curative in 1 and 5 died of postoperative complications. Thus, surgery is not safe in all cases. However, despite her age and bilateral renal dysfunction, our patient was successfully operated on.

Aged↗