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Biomedical subjects

M Matsuhashi

Publications and source records attributed to M Matsuhashi.

At least 55 records · Page 3Linked to original sources

New mutations fts-36, lts-33, and ftsW clustered in the mra region of the Escherichia coli chromosome induce thermosensitive cell growth and division.

Three new mutants of Escherichia coli showing thermosensitive cell growth and division were isolated, and the mutations were mapped to the mra region at 2 min on the E. coli chromosome map distal to leuA. Two mutations were mapped closely upstream of ftsI (also called pbpB), in a region of 600 bases; the fts-36 mutant showed thermosensitive growth and formed filamentous cells at 42 degrees C, whereas the lts-33 mutant lysed at 42 degrees C without forming filamentous cells. The mutation in the third new thermosensitive, filament-forming mutant, named ftsW, was mapped between murF and murG. By isolation of these three mutants, about 90% of the 17-kilobase region from fts-36-lts-33 to envA could be filled with genes for cell division and growth, and the genes could be aligned.

Base Sequence↗

Structural similarity among Escherichia coli FtsW and RodA proteins and Bacillus subtilis SpoVE protein, which function in cell division, cell elongation, and spore formation, respectively.

The Escherichia coli cell division gene ftsW (2 min) was cloned and sequenced. It encodes a hydrophobic protein(s) with 414 and/or 384 amino acid residues. The deduced amino acid sequence and the hydropathy profile of the protein showed high homology with those of the E. coli RodA protein functioning in determination of the cell shape and the Bacillus subtilis SpoVE protein functioning in spore formation. Probably similar functional membrane proteins are involved in these three cell cycle process.

Amino Acid Sequence↗

Inhibition of growth of ftsQ, ftsA, and ftsZ mutant cells of Escherichia coli by amplification of a chromosomal region encompassing closely aligned cell division and cell growth genes.

Amplification of a 2.6-kilobase chromosomal fragment of the mra region of Escherichia coli encompassing the ftsI(pbpB) gene and an open reading frame upstream with lethal to E. coli strains with mutations of the flanking cell division genes ftsQ, ftsA, and ftsZ. A shortened fragment in which the major portion of ftsI was deleted also had lethal effects on ftsQ and ftsZ mutants.

Bacterial Proteins↗

New mre genes mreC and mreD, responsible for formation of the rod shape of Escherichia coli cells.

New shape-determining genes in the mre cluster at 71 min on the Escherichia coli chromosome map, named mreC and mreD, were identified by complementation experiments using delta mre-678 mutant cells, which have a 5-kilobase-pair deletion encompassing the mre region, and by DNA sequencing. The delta mre-678 mutant cells required three genes, the previously reported mreB gene and the two new genes, to restore the normal rod shape of the cells and normal sensitivity of growth to mecillinam. The mreC gene is preceded by the mreB gene and by a 65-base-pair spacing sequence containing a palindrome sequence and a possible Shine-Dalgarno sequence. The deduced amino acid sequence of the MreC protein consists of 367 amino acid residues with a molecular weight of 39,530. The initiation codon of the mreD gene overlaps the termination codon of the mreC gene by one nucleotide residue. The deduced amino acid sequence of the MreD protein consists of 162 amino acid residues with a molecular weight of 18,755. In vitro, the coding frames of mreC and mreD produced proteins with Mrs of 40,000 and 15,000, respectively, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Amdinocillin↗

Expression and inducibility in Staphylococcus aureus of the mecA gene, which encodes a methicillin-resistant S. aureus-specific penicillin-binding protein.

A beta-lactam-sensitive strain of Staphylococcus aureus could be converted to methicillin resistance by the introduction of a plasmid carrying the 4.3-kilobase HindIII chromosomal DNA fragment which encoded the mecA gene from a methicillin-resistant S. aureus. Transformant cells produced methicillin-resistant S. aureus-specific penicillin-binding protein constitutively, and additional insertion of an inducible penicillinase plasmid caused production of the pencillin-binding protein to become inducible.

Bacterial Proteins↗

Negative control of cell division by mreB, a gene that functions in determining the rod shape of Escherichia coli cells.

Exponentially growing Escherichia coli cells containing additional copies of the shape-determining gene mreB were found to be elongated, whereas mreB mutant cells were spherical and overproduced penicillin-binding protein 3, a septum peptidoglycan synthetase. The effect of the mreB gene on expression of ftsI, the structural gene for penicillin-binding protein 3, was examined by using an ftsI-lacZ fusion gene on a plasmid. Formation of beta-galactosidase from the fusion gene was significantly increased in mreB129 mutant cells, and its overproduction was suppressed to a normal level by the presence of a plasmid containing the mreB gene. These results indicate a negative mechanism of control of cell division by this morphology gene and suggest that the gene functions in determining whether division or elongation of the cells occurs.

Cell Division↗

[A case of retroperitoneal abscess accompanying pregnancy].

A 40-year-old pregnant woman, who suffered a continuous episode of fever and anemia since the 26th week of pregnancy, was referred to our clinic for evaluation of her left flank pain and tumor. As retroperitoneal tumor was suspected by retrograde pyelography, sonography and CT, translumbar tumorectomy was performed under general anesthesia. The histological diagnosis was retroperitoneal abscess.

Abscess↗

[Clinical results of endourologic technique for upper urinary calculi].

Between May, 1985 and September, 1987, percutaneous nephrolithotripsy (PNL) and transurethral ureterolithotripsy (TUL) have been performed at our hospital and five affiliated hospitals. We report on the therapeutic results of 216 cases treated during this period. PNL and TUL were performed 168 times on 120 patients (119 males and 41 females) and 57 times on 56 patients (35 males and 21 females), respectively. Rate of success with PNL was 80.7% (103/130) for renal calculus, 85.7% (18/21) for upper ureteral calculus, and 47.1% (8/17) for middle-lower ureteral calculus. Rate of success with TUL was 57.9% (33/57). Of these, cases of ureteral lower edge calculus showed the lowest rate of success, being 33.3% (2/6). Complications of the cases undergoing PNL and TUL were: ureteral damage in 13; hemorrhage in 6; renal pelvic damage in 4; ureteral lower edge stenosis in 3; pyrexia in 3; cardiac insufficiency in 2 and retention of perfusate in posterior peritoneal cavity, pneumonia and renal insufficiency in 1 case each.

Adolescent↗

Primary carcinoma of seminal vesicle. Diagnosis assisted by sonography.

A case of primary carcinoma of the seminal vesicle in a nineteen-year-old man is described. This patient represents the twelfth case reported in the literature in Japan. The experience gained in this case indicated the potential usefulness of ultrasonography in the diagnosis of primary carcinoma of the seminal vesicle.

Adenocarcinoma, Papillary↗

Demonstration of varicella-zoster virus antigens in the vitreous aspirates of patients with acute retinal necrosis syndrome.

Four cases of acute retinal necrosis (ARN) syndrome were studied virologically. Varicella-zoster virus (VZV) antigen was demonstrated by immunofluorescence in cells from vitreous aspirates of two cases. No herpes simplex virus (HSV) or cytomegalovirus (CMV) antigens were detected by the same technique. Antibody to VZV in vitreous fluid was present in two cases; however, it was not detected in sera. Although virus isolation was unsuccessful, these findings strongly suggest that VZV may play an important role in the etiology of ARN syndrome.

Adult↗

Identification of the penicillin-binding active site of penicillin-binding protein 2 of Escherichia coli.

We determined the active site of penicillin-binding protein (PBP) 2 of Escherichia coli. A water-soluble form of PBP 2, which was constructed by site-directed mutagenesis, was purified by affinity chromatography, labeled with dansyl-penicillin, and then digested with a combination of proteases. The amino acid composition of the labeled chymotryptic peptide purified by HPLC was identical with that of the amino acid sequence, Ala-Thr-Gln-Gly-Val-Tyr-Pro-Pro-Ala-Ser330-Thr-Val-Lys-Pro (residues 321-334) of PBP 2, which was deduced from the nucleotide sequence of the pbpA gene encoding PBP 2. This amino acid sequence was verified by sequencing the labeled tryptic peptide containing the labeled chymotryptic peptide region. A mutant PBP 2 (thiol-PBP 2), constructed by site-directed mutagenesis to replace Ser330 with Cys, lacked the penicillin-binding activity. These findings provided evidence that Ser330 near the middle of the primary structure of PBP 2 is the penicillin-binding active-site residue, as predicted previously on the basis of the sequence homology. Around this active site, the sequence Ser-Xaa-Xaa-Lys was observed, which is conserved in the active-site regions of all E. coli PBPs so far studied, class A and class C beta-lactamases, and D-Ala carboxypeptidases. The COOH-terminal amino acid of PBP 2 was identified as His633.

Acyltransferases↗

Simultaneous recording of visual evoked potentials and electroretinograms in patients with unilateral opaque media.

We studied the correlation between preoperative visual evoked potentials (VEPs) and postoperative visual acuities in patients with unilateral opaque media. The fellow eye was used as the control. The filtering effect due to opaque media could be assessed on the basis of the difference of the electroretinogram (ERG) threshold in the two eyes. 'Modified VEPs' were then calculated by eliminating the assessed filtering effects, which improved our ability to detect the potential for visual recovery. In the normal eye, the stimulus threshold for VEP is lower than that for ERG by 2 or 3 log units. Therefore should a higher stimulus threshold be found for VEP than ERG, severe optic nerve damage is highly probable in eyes with media opacties.

Electroretinography↗

Determinations of the DNA sequence of the mreB gene and of the gene products of the mre region that function in formation of the rod shape of Escherichia coli cells.

The 6.5-kilobase mre region at 71 min in the Escherichia coli chromosome map, where genes involved in formation of a rod-shaped cell form a gene cluster, was analyzed by in vivo protein synthesis in a maxicell system and by base sequencing of DNA. An open reading frame that may code for a protein with an Mr of about 37,000 on sodium dodecyl sulfate-polyacrylamide gels was found and was correlated with the mreB gene. N-terminal amino acid sequencing of the hybrid mreB-lacZ protein confirmed the production by mreB of a protein of 347 amino acid residues with a molecular weight of 36,958. The amino acid sequence of this protein deduced from the DNA sequence showed close similarity with that of a protein of the ftsA gene which is involved in cell division of E. coli. Three other contiguous genes that formed three proteins with Mrs of about 40,000, 22,000, and 51,000, respectively, were detected downstream of the mreB gene by in vivo protein synthesis. The mreB protein and some of these three proteins may function together in determination of cell shape.

Amino Acid Sequence↗

Thermosensitive omsA mutation of Escherichia coli that causes thermoregulated release of periplasmic proteins.

A mutant of Escherichia coli with a thermosensitive defect, possibly in the outer membrane (omsA mutant), was isolated from E. coli K-12 by mutagenization and selection for thermosensitivity and beta-lactam supersensitivity of growth. The mutant also showed very high sensitivity to other antibiotics, such as macarbomycin, midecamycin, rifampin, and bacitracin. The mutation was recessive to the wild type and was mapped at about 4 min on the E. coli chromosome between fhuA and metD. The mutation caused rapid release into the medium of periplasmic enzymes such as RTEM penicillinase but practically no cytoplasmic enzyme when cells grown at 30 degrees C were transferred to 37 or 42 degrees C. Electron microscopic observations showed many large double-layered vesicles attached to the surface of cells incubated at 42 degrees C. We conclude that the mutant had a mutation that caused a temperature-dependent defect in the outer membrane structure or its assembly (named an oms mutation). The omsA mutant may be useful for production of periplasmic proteins, which it releases into the culture medium on shift up of temperature.

Bacterial Outer Membrane Proteins↗

Evolution of an inducible penicillin-target protein in methicillin-resistant Staphylococcus aureus by gene fusion.

A new beta-lactam-inducible penicillin-binding protein (PBP) that has extremely low affinity to penicillin and most other beta-lactam antibiotics has been widely found in highly beta-lactam(methicillin)-resistant Staphylococcus aureus (MRSA). The gene for this protein was sequenced and the nucleotide sequence in its promoter and close upstream area was found to show close similarity with that of staphylococcal penicillinase, while the amino acid sequence over a wide range of the molecule was found to be similar to those of two PBPs of Escherichia coli, the shape-determining protein (PBP 2) and septum-forming one (PBP 3). Probably the MRSA PBP (Mr 76462) evolved by recombination of two genes: an inducible type I penicillinase gene and a PBP gene of a bacterium, causing the formation of a beta-lactam-inducible MRSA PBP.

Anti-Bacterial Agents↗

Goldmann-Favre syndrome in a four-year-old-girl.

We report a suspected early stage of Goldmann-Favre syndrome in a four-year-old girl complaining of poor visual acuity in both eyes. Ophthalmological examinations were performed including fundus examination, fluorescein angiography, and electrophysiological studies. However, dark adaptation and visual field testing were not performed because of her age. Ophthalmoscopic examinations revealed peripheral retinoschisis in the superior temporal retina, diffusely abnormal retinal pigment epithelium, and lack of macular and foveal reflexes in both eyes. Foveal retinoschisis was not observed and the vitreous did not show severe degeneration. Fluorescein angiography showed many leaking points posterior to the retinoschisis. Scotopic electroretinogram (ERG) was almost non-recordable. Photopic electroretinogram showed a marked reduction of the b-wave with a negative shape and delayed peak time. Flash visual evoked potentials were within normal range. It is very difficult to distinguish this case from X-linked juvenile retinoschisis by ophthalmological findings. However, Goldmann-Favre syndrome is most suspected from the fact that the patient is female and her parents and brothers are normal.

Child, Preschool↗

Familial retinoschisis in female patients.

We report two female patients, a mother and daughter, with bilateral foveal changes that resembled those of X-linked recessive juvenile retinoschisis. The 23-year-old daughter had flat retinoschisis at the temporal periphery with multiple small inner-layer breaks in both eyes. There was foveal retinoschisis with fine radial folds. The optic disc was dragged to the nasal side. The 49-year-old mother also had foveal retinoschisis in each eye but there was no peripheral retinoschisis. In the left eye several retinal breaks with minimal retinal detachment were found. Electrophysiological findings in both cases were similar. Single-flash electroretinogram (ERG) showed normal a-wave and decreased b-wave, presenting a negative shape. Averaged scotopic and photopic ERGs showed slightly reduced b-waves, but they were within normal ranges. Visually evoked potentials were subnormal. Ophthalmoscopic and electrophysiologic findings were compatible with X-linked recessive juvenile retinoschisis, but an autosomal dominant inheritance was most likely. Our cases do not follow previously reported characteristics and may represent a new clinical entity.

Adult↗