Biomedical subjects

K Ochi

Publications and source records attributed to K Ochi.

At least 19 recordsLinked to original sources

PubMed2004

Listeria monocytogenes isolated from cold-smoked fish products in Osaka City, Japan.

Listeria monocytogenes contamination of ready-to-eat seafood products commercially available in Osaka was examined between 1999 and 2000. L. monocytogenes was isolated from 12 (13%) of the 95 products tested. All positive samples were from cold-smoked fish with 9 being obtained during the summer. Thirteen isolates of L. monocytogenes were typed by pulsed-field gel electrophoresis (PFGE) and polymerase chain reaction (PCR)-based typing methods. Isolates of the same serotype originating from the same manufacturer gave similar DNA profiles, irrespective of the type of sample or date of isolation. The finding suggest that persistent strains in each manufacturing facility proliferate during the summer and contaminate products during manufacturing processes.

Animals↗

PubMed2004

The novel mutation K87E in ribosomal protein S12 enhances protein synthesis activity during the late growth phase in Escherichia coli.

Resistance to streptomycin in bacterial cells often results from a mutation in the rpsL gene that encodes the ribosomal protein S12. We found that a particular rpsL mutation (K87E), newly identified in Escherichia coli, causes aberrant protein synthesis activity late in the growth phase. While protein synthesis decreased with age in cells in the wild-type strain, it was sustained at a high level in the mutant, as determined using living cells. This was confirmed using an in vitro protein synthesis system with poly(U) and natural mRNAs (GFP mRNA and CAT mRNA). Other classical rpsL mutations (K42N and K42T) tested did not show such an effect, indicating that this novel characteristic is typical of ribosomes bearing the K87E mutant form of S12, although the K87E mutation conferred the streptomycin resistance and error-restrictive phenotypes also seen with the K42N and K42T mutations. The K87E (but not K42N or K42T) mutant ribosomes exhibited increased stability of the 70S complex in the presence of low concentrations of magnesium. We propose that the aberrant activation of protein synthesis at the late growth phase is caused by the increased stability of the ribosome.

Chloramphenicol O-Acetyltransferase↗

PubMed2002

A rifampicin resistance mutation in the rpoB gene confers ppGpp-independent antibiotic production in Streptomyces coelicolor A3(2).

In Streptomyces coelicolor A3(2), deletion of relA or a specific mutation in rplK ( relC) results in an inability to synthesize ppGpp (guanosine 5'-diphosphate 3'-diphosphate) and impairs production of actinorhodin. We have found that certain rifampicin-resistant ( rif) mutants isolated from either relA or relC strains regain the ability to produce actinorhodin at the same level as the wild-type strain, although their capacity to synthesize ppGpp is unchanged. These rif mutants were found to have a missense mutation in the rpoB gene that encodes the RNA polymerase beta-subunit. This rpoB mutation was shown to be responsible for the observed changes in phenotype, as demonstrated by gene replacement experiments. Gene expression analysis revealed that the restoration of actinorhodin production in both relA and relC strains is accompanied by increased expression of the pathway-specific regulator gene actII-ORF4, which is normally decreased in the rel mutants. In addition to the restoration of antibiotic production, the rif mutants also exhibited a lower rate of RNA synthesis compared to the parental strain when grown in a rich medium, suggesting that these mutant RNA polymerases behave like "stringent" RNA polymerases. These results indicate that rif mutations can alter gene expression patterns independently of ppGpp. We propose that RNA polymerases carrying particular rif mutations in the beta-subunit can functionally mimic the modification induced by binding of ppGpp.

Amino Acid Sequence↗

PubMed2001

The inhibitory effect of the autoantigen La on in vitro 3' processing of mammalian precursor tRNAs.

Mammalian tRNA 3' processing endoribonuclease (3' tRNase) can remove a 3' trailer from various precursor (pre)-tRNAs. We investigated what effect the autoantigen La has on 3' processing, since the La protein is known to bind to a 3'-terminal uridine tract of pre-tRNAs. We tested sixteen different pre-tRNA(Arg) substrates containing various 3' trailers with or without a 5' leader sequence for in vitro processing by pig 3' tRNase, and for gel-retardation in the presence or absence of human La protein. The R-TUUU series consists of four pre-tRNAs containing 6, 8, 11 and 15 nt 3' trailers ending with UUU and no 5' leader, while the R-TAGC series consists of the same four pre-tRNAs as R-TUUU except that the terminal sequence is AGC. The R-6LTUUU and R-6LTAGC series are derived from R-TUUU and R-TAGC, respectively, by adding a 6 nt 5' leader. La differentially inhibited their processing and bound to the pre-tRNAs; the 50 % inhibitory concentrations for the R-TUUU, R-TAGC, R-6LTUUU, and R-6LTAGC series were 82 to >850, >850, 2 to 292 and 573 to 785 nM, respectively, and the dissociation constants were 10 to 840, >850, 3 to 203 and 155 to 520 nM, respectively. These results indicate that both the terminal sequence UUU and the 5' leader contribute to more severe inhibition of 3' processing via tighter interaction with La. With respect to the R-TUUU and R-6LTUUU series, on the whole, the La inhibition was enhanced as the 3' trailer lengths decreased. Taken together, our results suggest that the La protein sterically hinders 3' tRNase from binding a pre-tRNA molecule probably near the cleavage site.

Animals↗

PubMed2001

Electrocochleogram after transection of vestibulo-cochlear nerve in a patient with a large acoustic neurinoma.

This study reports pre- and post-operative compound action potentials (CAPs) that were recorded from a 27-year-old woman with an acoustic neurinoma. During surgery it was necessary to totally sever her vestibulo-cochlear nerve to excise a large tumor. A pure tone audiogram changed to the scale-out pattern immediately after operation. However, CAP, the waveform of which was broadened, has been recorded 3 years post-operatively with a threshold elevation of 10 dB over the pre-operative threshold. This phenomenon suggests that CAP may originate from the extreme periphery of the auditory nerve within the cochlea. The broadening of the CAP was assumed to result from enhancement of the negative summating potential included in the CAP. We studied the effect of preceding stimulus on CAP using paired click stimuli pre- and post-operatively. A reduction of CAP amplitude in response to the second click of paired clicks was markedly suppressed in the inter-click interval between 3 and 80 ms post-operatively. We speculate that depletion of adaptation induced the abnormal CAP recovery described above and that the lateral efferent nerve system was involved in abnormal CAP adaptation with transection of the vestibulo-cochlear nerve in this case.

Action Potentials↗

PubMed2001

Condylar bony change and self-reported parafunctional habits in prospective orthognathic surgery patients with temporomandibular disorders.

OBJECTIVE: The purpose of this study was to investigate the relationship between self-reported parafunctional habits and condylar bony change and disk displacement in orthognathic surgery patients with signs and symptoms of temporomandibular joint disorders. STUDY DESIGN: This is a cross-sectional retrospective study of pretreatment helical computed tomography scans and questionnaires of 94 female orthognathic surgery patients. RESULTS: Condylar bony change, unilaterally or bilaterally, was found in 56.4% of the subjects, or 43.6% of the joints. Disk displacement, unilaterally or bilaterally, was seen in 59.6% of the subjects, or 45.7% of the joints. Bruxism and clenching was significantly associated with condylar bony change and disk displacement. Subjects with 3 or more parafunctional habits showed a significantly higher rate of bilateral condylar bony change. CONCLUSION: Our results suggest that bruxism and clenching might be related to deterioration of the temporomandibular joint and that the greater the number of parafunctional habits a subject has, the higher the risk of condylar bony change.

Bruxism↗

PubMed2001

Variance of vestibular-evoked myogenic potentials.

OBJECTIVES/HYPOTHESIS: Vestibular-evoked myogenic potential (VEMP) has been thought to originate from sacculus. The variance of this potential and the effectiveness of the adjustments of pInII amplitudes using average muscle tonus of ipsilateral sternocleidomastoid muscle were evaluated. In addition, clinical application of VEMP was examined in patients with acoustic tumors (ATs) and vestibular neurolabyrinthitis (VNL). STUDY DESIGN: Prospective evaluation of the VEMP in 18 normal volunteers and 6 patients. METHODS: Variance and left-right difference of each parameter, including pI latency, nII latency, pInII amplitude, and threshold, was analyzed. Input-output function of pInII amplitude was evaluated. Average muscle tonus was calculated in 20 ears and applied for adjustment of pInII amplitude. Sensitivity of each parameter of VEMP was examined in 3 patients with ATs and 3 patients with VNL. RESULTS: VEMP was present in all 36 ears of 18 control subjects. Thresholds of VEMP for normal subjects were 80 to 95 dB normal hearing level (nHL). The muscle tonus affected pInII amplitude significantly; however, no statistically significant improvement was observed in test-retest investigation after adjustment using muscle tonus. The threshold of the affected side was elevated compared with the non-affected side in all patients with ATs, whereas 2 of 3 patients showed normal pInII-ratio. One patient with VNL presented normal VEMP, whereas 2 patients presented no VEMP to the highest stimulus intensity. CONCLUSIONS: Interaural difference of thresholds might be the most useful parameters. Adjustment using average muscle tonus is not necessary when the subject is able to get sufficient muscle tonus.

Acoustic Stimulation↗

PubMed2001

Sound-evoked myogenic potentials and responses with 3-ms latency in auditory brainstem response.

OBJECTIVES: The vestibular-evoked myogenic potential (VEMP) has shed new light on vestibular testing. A large negative deflection with a 3-ms latency within the auditory brainstem response (ABR) has been reported in some patients with deafness. This negative deflection has been termed the N3 potential and it is assumed to be a vestibular-evoked potential. This study investigated the relationship between the VEMP and the N3 potential. STUDY DESIGN: Prospective evaluation of the VEMP and the N3 potential in 21 patients. METHODS: The oto-neurological tests, including caloric test, hearing sensitivity test, VEMP, and ABR, were performed and data were analyzed. RESULTS: The average hearing threshold ranged from 65 to above 110 dB, which includes 9 (37.5%) totally deaf ears. The N3 potentials were recorded in 10 (41.7%) ears. A normal VEMP was detected in 16 (66.7%) ears. Canal paresis was observed in 11 (45.8%) ears. CONCLUSIONS: Both the VEMP and the N3 potential appear to originate from the sacculus, but because the characteristics of these two responses are not identical, additional factors might be involved in the generation of the N3 potential.

Acoustic Stimulation↗

PubMed2001

Study on free radicals and pancreatic fibrosis--pancreatic fibrosis induced by repeated injections of superoxide dismutase inhibitor.

The exact mechanisms of the development of pancreatic fibrosis are still unknown. To clarify the relationship between pancreatic fibrosis and free radicals, the effect of the administration of a superoxide dismutase (SOD) inhibitor, diethyldithiocarbamate (DDC), on pancreatic fibrosis in rats was studied. A single intraperitoneal injection of 500 mg/kg of DDC significantly reduced SOD activity and significantly increased lipid peroxidation products in the pancreas, showing no histologic changes of inflammation or necrosis. Repeated administration of 500 mg/kg DDC, twice a week, caused inter- and intralobular fibrosis with atrophy of acinar cells in the pancreas for at least 2 weeks without fibrosis of the liver and kidney. Administration of allopurinol showed preventive effects against DDC-induced pancreatic fibrosis. In conclusion, repeated administration of DDC, which caused pancreatic fibrosis, is a new experimental model of pancreatic fibrosis from the viewpoint of oxidative stress.

Allopurinol↗

PubMed2001

Novel approach for improving the productivity of antibiotic-producing strains by inducing combined resistant mutations.

We developed a novel approach for improving the production of antibiotic from Streptomyces coelicolor A3(2) by inducing combined drug-resistant mutations. Mutants with enhanced (1.6- to 3-fold-higher) actinorhodin production were detected at a high frequency (5 to 10%) among isolates resistant to streptomycin (Str(r)), gentamicin (Gen(r)), or rifampin (Rif(r)), which developed spontaneously on agar plates which contained one of the three drugs. Construction of double mutants (str gen and str rif) by introducing gentamicin or rifampin resistance into an str mutant resulted in further increased (1.7- to 2.5-fold-higher) actinorhodin productivity. Likewise, triple mutants (str gen rif) thus constructed were found to have an even greater ability for producing the antibiotic, eventually generating a mutant able to produce 48 times more actinorhodin than the wild-type strain. Analysis of str mutants revealed that a point mutation occurred within the rpsL gene, which encodes the ribosomal protein S12. rif mutants were found to have a point mutation in the rpoB gene, which encodes the beta-subunit of RNA polymerase. Mutation points in gen mutants still remain unknown. These single, double, and triple mutants displayed in hierarchical order a remarkable increase in the production of ActII-ORF4, a pathway-specific regulatory protein, as determined by Western blotting analysis. This reflects the same hierarchical order observed for the increase in actinorhodin production. The superior ability of the triple mutants was demonstrated by physiological analyses under various cultural conditions. We conclude that by inducing combined drug-resistant mutations we can continuously increase the production of antibiotic in a stepwise manner. This new breeding approach could be especially effective for initially improving the production of antibiotics from wild-type strains.

Anthraquinones↗

PubMed2001

Construction of an in vivo nonsense readthrough assay system and functional analysis of ribosomal proteins S12, S4, and S5 in Bacillus subtilis.

To investigate the function of ribosomal proteins and translational factors in Bacillus subtilis, we developed an in vivo assay system to measure the level of nonsense readthrough by utilizing the LacZ-LacI system. Using the in vivo nonsense readthrough assay system which we developed, together with an in vitro poly(U)-directed cell-free translation assay system, we compared the processibility and translational accuracy of mutant ribosomes with those of the wild-type ribosome. Like Escherichia coli mutants, most S12 mutants exhibited lower frequencies of both UGA readthrough and missense error; the only exception was a mutant (in which Lys-56 was changed to Arg) which exhibited a threefold-higher frequency of readthrough than the wild-type strain. We also isolated several ribosomal ambiguity (ram) mutants from an S12 mutant. These ram mutants and the S12 mutant mentioned above (in which Lys-56 was changed to Arg) exhibited higher UGA readthrough levels. Thus, the mutation which altered Lys-56 to Arg resulted in a ram phenotype in B. subtilis. The efficacy of our in vivo nonsense readthrough assay system was demonstrated in our investigation of the function of ribosomal proteins and translational factors.

Amino Acid Substitution↗

PubMed2001

Molecular and functional analyses of the gene (eshA) encoding the 52-kilodalton protein of Streptomyces coelicolor A3(2) required for antibiotic production.

Analysis of proteins recovered in the S100 precipitate fraction of Streptomyces griseus after ultracentrifugation led to the identification of a 52-kDa protein which is produced during the late growth phase. The gene (eshA) which codes for this protein was cloned from S. griseus, and then its homologue was cloned from Streptomyces coelicolor A3(2). The protein was deduced to be 471 amino acids in length. The protein EshA is characterized by a central region that shows homology to the eukaryotic-type cyclic nucleotide-binding domains. Significant homology was also found to MMPI in Mycobacterium leprae, a major antigenic protein to humans. The eshA gene mapped near the chromosome end and was not essential for viability, as demonstrated by gene disruption experiments, but its disruption resulted in the abolishment of an antibiotic (actinorhodin but not undecylprodigiosin) production. Aerial mycelium was produced as abundantly as by the parent strain. Expression analysis of the EshA protein by Western blotting revealed that EshA is present only in late-growth-phase cells. The eshA gene was transcribed just preceding intracellular accumulation of the EshA protein, as determined by S1 nuclease protection, indicating that EshA expression is regulated at the transcription level. The expression of EshA was unaffected by introduction of the relA mutation, which blocks ppGpp synthesis.

Amino Acid Sequence↗

PubMed2001

Complete nucleotide sequence of the cytochrome b gene of channel catfish Ictalurus punctatus and comparison of sequence homology among channel catfish and other fishes.

To clarify the phylogenic relationship of channel catfish with other fishes, the cytochrome b (Cyt b) gene of the catfish was cloned and sequenced. Channel catfish (Ictalurus punctatus) belonging to the family Ictaluridae in the order Siluriformes showed a 78.4-87.4% similarity to all but one fish of the family Cyprinidae and river loach Crossostoma lacustre of the family Balitoridae in the order Cypriniformes in which genes had already been sequenced, and a 97.2% similarity to the goldfish (Carassius auratus) belonging to the family Cyprinidae. Within the family Cyprinidae, a 78.8-89.2% similarity to one another was recorded. In addition, the similarity rate between the family Cyprinidae and the family Balitoridae reached a value of 77.8-79.9% in the order Cypriniformes. Furthermore, in an unrooted phylogenetic tree consisting of four branches among eight fishes, channel catfish and goldfish appeared in the same branch. These results suggested that the Cyt b gene of the channel catfish in the order Siluriformes was closely related to that of a goldfish in the order Cypriniformes. The results were not agreement with the morphological classification. Genetic reclassification of the fishes may be necessary to identify the ancestor. This is the first report on the cloning and complete sequencing the Cyt b gene of the channel catfish which may contribute to the genetic reclassification of catfishes belonging to the order Siluriformes.

Amino Acid Sequence↗

PubMed2001

Giant negative T waves during interferon therapy in a patient with chronic hepatitis C.

Interferon-alpha (IFN-alpha) has been widely used for treatment of chronic hepatitis C in Japan. In general, cardiovascular adverse reactions are rare in association with IFN-alpha therapy. Here, a 64-year-old man with chronic active hepatitis C complained of fatigue, palpitation and depression, and developed atrial fibrillation with prominent negative T waves during IFN-alpha therapy. Echocardiogram showed septal and apical hypertrophy. Three days after discontinuation of IFN-alpha, subjective symptoms and atrial fibrillation subsided. It is unclear whether or not IFN-alpha induced the giant negative T waves with apical hypertrophy. We might observe the developing course of hepatitis C virus (HCV)-related myocardial hypertrophy by chance. Cardiovascular toxicity should be carefully monitored during IFN-alpha therapy even in patients with minor cardiac disease, such as premature ventricular contracture (PVC) and mild hypertension.

Antihypertensive Agents↗

PubMed2001

[The clinical application and modification of the Quad Helix appliance].

OBJECTIVE: The aims of this study are to investigate the working mechanism, the characteristics, the clinical application and the suggested modification of the Quad Helix appliance. METHODS: A 7-year-old, female patient with Pierre-Rokin syndrome, who was preformed with palatorrhaphy at 21-month-old treated by using a Quad Helix appliance for one year. RESULTS: After one-year treatment, the wide between the maxillary first molars increased 9.65 mm, and the wide between maxillary canines increased 5.20 mm. The wide between the mandibular first molars also increased 3.60 mm, however the wide between mandibular canines decreased 5.20 mm. CONCLUSION: The Quad-helix appears to be a successful Orthodontic appliance to expand the narrow maxillary or mandibular arches.

Child↗

PubMed2000

Multinucleated giant cells in submucosal layer of human urinary bladder: an immunohistochemical and electron microscopic study.

Multinucleated giant cells (MGC) detected in the submucosal layer of human urinary bladder mainly associated with transitional cell carcinoma were examined immunohistochemically and ultrastructurally. The cases examined totaled 29, namely 14 cases with transitional cell carcinoma and another 15 cases mostly with malignancy in other organs. Histologically, MGC were smooth, irregular or dendritic in shape, and tended to increase in number in the vicinity of cancer or marked inflammation. They were consistently positive for not only vimentin, but also MB-2, and CD34, and were mostly positive for proliferating cell nuclear antigen (PCNA), but not MIB-1 (Ki-67) and HLA-DRalpha antigens. On occasion, antibodies to alpha-smooth muscle actin (alpha-SMA), muscle actin (M-actin), CD68 (KP-1) and alpha subunit of S-100 protein also yielded positive reactions. Interestingly, aggregated short bulbous processes were ultrastructurally observed on their surface in parts. These findings suggested that MGC in the submucosal layer of human urinary bladder were MB-2 and CD34-positive multipotential mesenchymal cells with no mitotic activity expressing fibroblastic (vimentin), myofibroblastic (alpha-SMA), or histiocytic (CD68) markers mostly in the vicinity of malignancy, and that these MGC were formed by fusion of mononuclear cells expressing identical markers with those of MGC. Further investigations are needed to clarify the exact function of MGC in human urinary bladder.

Adult↗

PubMed2000

Functional coupling of Ca(2+) channels to ryanodine receptors at presynaptic terminals. Amplification of exocytosis and plasticity.

Ca(2+)-induced Ca(2+) release (CICR) enhances a variety of cellular Ca(2+) signaling and functions. How CICR affects impulse-evoked transmitter release is unknown. At frog motor nerve terminals, repetitive Ca(2+) entries slowly prime and subsequently activate the mechanism of CICR via ryanodine receptors and asynchronous exocytosis of transmitters. Further Ca(2+) entry inactivates the CICR mechanism and the absence of Ca(2+) entry for >1 min results in its slow depriming. We now report here that the activation of this unique CICR markedly enhances impulse-evoked exocytosis of transmitter. The conditioning nerve stimulation (10-20 Hz, 2-10 min) that primes the CICR mechanism produced the marked enhancement of the amplitude and quantal content of end-plate potentials (EPPs) that decayed double exponentially with time constants of 1.85 and 10 min. The enhancement was blocked by inhibitors of ryanodine receptors and was accompanied by a slight prolongation of the peak times of EPP and the end-plate currents estimated from deconvolution of EPP. The conditioning nerve stimulation also enhanced single impulse- and tetanus-induced rises in intracellular Ca(2+) in the terminals with little change in time course. There was no change in the rate of growth of the amplitudes of EPPs in a short train after the conditioning stimulation. On the other hand, the augmentation and potentiation of EPP were enhanced, and then decreased in parallel with changes in intraterminal Ca(2+) during repetition of tetani. The results suggest that ryanodine receptors exist close to voltage-gated Ca(2+) channels in the presynaptic terminals and amplify the impulse-evoked exocytosis and its plasticity via CICR after Ca(2+)-dependent priming.

Animals↗

PubMed2000

Polyketide synthase gene coupled to the peptide synthetase module involved in the biosynthesis of the cyclic heptapeptide microcystin.

The peptide synthetase gene operon, which consists of mcyA, mcyB, and mcyC, for the activation and incorporation of the five amino acid constituents of microcystin has been identified [T. Nishizawa et al. (1999) J. Biochem. 126, 520-529]. By sequencing an additional 34 kb of DNA from microcystin-producing Microcystis aeruginosa K-139, we identified the residual microcystin synthetase gene operon, which consists of mcyD, mcyE, mcyF, and mcyG, in the opposite orientation to the mcyABC operon. McyD consisted of two polyketide synthase modules, and McyE contained a polyketide synthase module at the N-terminus and a peptide synthetase module at the C-terminus. McyF was found to exhibit similarity to amino acid racemase. McyG consisted of a peptide synthetase module at the N-terminus and a polyketide synthase at the C-terminus. The microcystin synthetase gene cluster was conserved in another microcystin-producing strain, Microcystis sp. S-70, which produces Microcystin-LR, -RR, and -YR. Insertional mutagenesis of mcyA, mcyD, or mcyE in Microcystis sp. S-70 abolished microcystin production. In conclusion, the mcyDEFG operon is presumed to be responsible for 3-amino-9-methoxy-2,6, 8-trimethyl-10-phenyldeca-4,6-dienoic acid (Adda) biosynthesis, and the incorporation of Adda and glutamic acid into the microcystin molecule.

Amino Acid Sequence↗