PubMed Health⌕ Search

Biomedical subjects

H Fukuhara

Publications and source records attributed to H Fukuhara.

At least 55 records · Page 3Linked to original sources
PubMed1995

Linear mitochondrial DNAs from yeasts: telomeres with large tandem repetitions.

The terminal structure of the linear mitochondrial DNA (mtDNA) from the yeast Candida parapsilosis was investigated. This mtDNA, 30 kb long, has symmetrical ends forming inverted terminal repeats. These repeats are made up of a variable number of tandemly repeating units of 738 bp each; the terminal nucleotide corresponds to a precise position within the last repeat unit sequence. The ends had an open structure accessible to enzymes, with a 5' single-stranded extension of about 110 nucleotides. No circular forms were detected in the DNA preparations. Two other unrelated species, Pichia philodendra and Candida salmanticensis also appear to have a linear mtDNA of similar organization. These linear DNAs (which we name Type 2 linear mtDNAs) are distinct from the previously described linear mtDNAs of yeasts whose termini are formed by a closed hairpin loop (Type 1 linear mtDNA). The terminal structure of C. parapsilosis mtDNA is reminiscent of the linear mitochondrial genomes of the ciliate Tetrahymena although, in the latter, the telomeric tandem repeat unit is considerably shorter.

Base Sequence↗
PubMed1995

The respiratory system of Kluyveromyces lactis escapes from HAP2 control.

A functional homolog of the Saccharomyces cerevisiae HAP2 gene, coding for one element of a transcriptional activator complex, was cloned from the yeast Kluyveromyces lactis and its nucleotide sequence was determined. Inactivation of the gene had no significant effect on respiration-dependent growth, suggesting that the HAP2/3/4 complex has no major control over the formation of the mitochondrial respiratory system in K. lactis.

Amino Acid Sequence↗
PubMed1995

[Identification of Legionella pneumophila strains by polymerase chain reaction method which isolated from cooling tower water].

We studied the polymerase chain reaction (PCR) method with 5 strains of Legionella pneumophila serogroup 4, 7 strains of L. pneumophila serogroup 5, 7 strains of L. pneumophila UT and 15 strains of Legionella like organisms (LLO). We used mip-primers reported by Mahbubani et al and LEG-primers reported by Starnbach et al. Positive PCR results were obtained by all strains of L. pneumophila serogroup 4, L. pneamophila serogroup 5, L. pneumophila UT and 11 strains of LLO using mip-primers. Using LEG-primers, positive results were obtained by 2, 6, 5 and 8 strains respectively. All strains that showed positive results using LEG-primer PCR, were contained by the strains that showed positive results using mip-primer PCR. Conversely, there were some strains that were positive in mip-primer PCR and negative in LEG-primer simultaneously. Therefore, mip-primers is recommended to detect L. pneumophila in clinical specimens by PCR method.

Base Sequence↗
PubMed1995

[Liver herniation simulating a benign lung tumor].

A 49-year-old woman complained of coughing and sputum production a mass shadow was seen in the right lower lung field on a chest radiography. The shadow was first suspected to represent a benign lung tumor, because of its clear margin and narrow pedicle. After detailed examination liver herniation was suspected, since the CT number of the tumor was similar to that of the liver. Liver herniation was finally confirmed by chest MRI, liver scintigraphy and angiography of the abdomen. Liver herniation should be differentiated from an intrathoracic mass in the right lower lung field.

Diagnosis, Differential↗
PubMed1994

Creation of a functional promoter by rearrangement in a Kluyveromyces lactis linear plasmid.

The expression of genes from cytoplasmic killer plasmids in the yeast Kluyveromyces lactis depends on their own specific transcription system. Therefore, the kanamycin/G418-resistance-encoding gene, KmR, under its natural promoter cannot be expressed when integrated into the pGKL1 plasmid. However, one G418R transformant clone was isolated. The resistance was due to the presence of two modified plasmids, k1-kan2a (10.4 kb) and k1-kan2b (5.2 kb) which were derivatives of pGKL1 containing the KmR gene. In these mutant plasmids, a large part of pGKL1 has been replaced by the KmR gene harboring a rearranged 5'-flanking region extending over 600 bp. This new DNA sequence has been cloned and sequenced. The rearranged sequence allowed the KmR gene to be expressed at high level, enabling the transformant cells to grow on a medium containing G418 at 2 mg/ml. This high level of resistance was found to be due to increased transcription of the KmR gene. Primer extension experiments suggested that the rearranged upstream region of KmR contained transcription promoting sites recognized by the killer-plasmid-specific transcription system.

Amino Acid Sequence↗
PubMed1994

Complete DNA sequence of yeast chromosome XI.

The complete DNA sequence of the yeast Saccharomyces cerevisiae chromosome XI has been determined. In addition to a compact arrangement of potential protein coding sequences, the 666,448-base-pair sequence has revealed general chromosome patterns; in particular, alternating regional variations in average base composition correlate with variations in local gene density along the chromosome. Significant discrepancies with the previously published genetic map demonstrate the need for using independent physical mapping criteria.

Base Sequence↗
PubMed1994

Mitochondrial transfer RNA genes of the yeast Candida parapsilosis.

Fifteen tRNA-encoding genes were mapped on the linear mitochondrial (mt) DNA of the yeast, Candida parapsilosis, and their sequences determined. The gene order and gene sequences indicate that the mt genome of this yeast belongs to a group clearly different from the already known group of linear mt DNAs of yeast.

Base Sequence↗
PubMed1994

Genes of the linear mitochondrial DNA of Williopsis mrakii: coding sequences for a maturase-like protein, a ribosomal protein VAR1 homologue, cytochrome oxidase subunit 2 and methionyl tRNA.

The mitochondrial DNA (mtDNA) in some yeasts has a linear structure with inverted terminal repeats closed by a single-stranded loop. These mtDNAs have generally a constant gene order, beginning with a small ribosomal RNA gene at the right end and terminating with a cytochrome oxidase subunit 2 gene (COX2) at the left end, independently of the wide variation in genome size. In the mtDNAs from several species of the genus Williopsis, we found an additional open reading frame, ORF1, which was homologous to the Saccharomyces cerevisiae RF1 gene encoding a group I intron maturase-like protein. ORF1 genes from W. mrakii and W. suaveolens were mapped and sequenced. Next to ORF1, COX2 and methionyl tRNA genes were present on the opposite strand. The same relative positions of genes in the mtDNAs so far examined suggests that the constancy of gene order is generally conserved also at the level of individual tRNA genes. We identified another open reading frame, ORF2, in W. mrakii mtDNA. It was mapped next to the cytochrome oxidase subunit 3 gene. Rich in adenine-thymine bases, ORF2 appears to be a homologue of the VAR1 gene which codes for a small ribosomal subunit protein in S. cerevisiae mitochondria. Nucleotide sequences data have been deposited in the EmBL data library under the following Accession Numbers: X66594 (Apocytochrome b and ORF2 genes of W. mrakii), X66595 (ORF1, tRNA-Met and COX2 genes of W. mrakii), X73415 (tRNA-Met and COX2 genes of W. suaveolens), X73416 (ORF1 gene of W. suaveolens) and X73414 (tRNA-Met and COX2 genes of P. jadinii).

Amino Acid Sequence↗
PubMed1994

Linear DNA plasmids from Pichia etchellsii, Debaryomyces hansenii and Wingea robertsiae.

Linear DNA plasmids were found in the following yeasts: four strains of Kluyveromyces lactis, one of Debaryomyces hansenii, one of Wingea robertsiae and four of Pichia etchellsii. In each case, the plasmids were present as a pair of DNA molecules of different sizes. The plasmids of K. lactis strains were associated with a killer activity and their structure was similar to the known killer plasmids pGKL1 and 2. The plasmids from the other three species were different from pGKL plasmids and showed no killer activity against the yeast species tested so far. In all cases, the linear molecules possessed terminal (probably inverted) repeats and their 5' ends had a protected structure insensitive to lambda exonuclease, while the 3' ends were accessible to exonuclease III. All these strains could be efficiently cured of the plasmids by ultraviolet irradiation. The plasmids from D. hansenii (pDH1A and B) and from W. robertsiae (pWR1A and B) shared related sequences with some of the K. lactis killer plasmid genes (encoding the supposed DNA polymerases, RNA polymerase and the chitinase), suggesting related genome organization of these plasmids. The pair of plasmids from P. etchellsii (pPE1A and B) appear to be a distantly related member of the group. This pair showed no sequence homology with other plasmids, except weak homology with the putative RNA polymerase gene of pGKL2. None of the plasmids contained the sequences homologous to ORF3 and ORF4 of pGKL1 encoding the toxin resistance determinant and the toxin gamma subunit, respectively.

DNA, Fungal↗
PubMed1994

[Serodiagnosis of Legionella pneumonia--data of our laboratory in the recent 3 years].

We had examined antibody titers against Legionella spp. of patients' sera which were mainly sent from other hospitals, performed with the indirect fluorescent antibody (IFA) method. The clinical status of the cases diagnosed as Legionella pneumonia serologically, were also studied. Out of 105 cases with clinically suspected Legionella pneumonia, 15 cases (14.3%) were seropositive. In 9 out of the 15 cases (60.0%) were caused by Legionella pneumophila serogroup 1. Clinical outline of these 15 cases did not contradict those reported in the literature, and erythromycin was effective in many cases. Significant rises (more than four times) of the titer were observed 3 to 4 weeks after onset in most of cases. We would like to emphasize that this should be performed serodiagnosis of Legionella pneumonia.

Adult↗
PubMed1994

[Usefulness of polymerase chain reaction method in the early diagnosis of Pneumocystis carinii pneumonia].

We evaluated the sensitivity and specificity of polymerase chain reaction (PCR) method in the detection of Pneumocystis carinii, and compared its clinical usefulness with that of a previous method, toluidine blue-O (TBO) staining in diagnosis of P. carinii pneumonia (PCP) using clinical samples such as sputum and bronchoalveolar lavage fluid (BALF). Using P. carinii obtained from BALF of the infected rats, we showed that PCR showed 100 fold more sensitivity than TBO staining as indicated by the comparison of their detection limits (78 cysts/ml vs 7800 cysts/ml). In addition, the detection using PCR in this study was specific for P. carinii because all other pathogens examined were not positive. In the clinical study we analyzed 44 patients including 10 with definite PCP, 8 with portable PCP and 26 with other diagnosis. In the sputum, the sensitivity and specificity were 100% and 95%, respectively, in PCR, while 13% and 100% in TBO staining. These results revealed that PCR was much superior to TBO staining in sensitivity especially when the sputum was used as samples, and suggested that examination of the sputum by PCR was clinically very useful in early diagnosis of P. carinii pneumonia.

Adult↗
PubMed1994

[A case report of miliary tuberculosis with acute respiratory failure diagnosed by polymerase chain reaction method].

A 51-year-old male was admitted because of cough and high fever. His chest X-ray and CT revealed miliary shadows, he was administered some antibiotics. But on the 8th day, his chest X-ray revealed ground glass shadows and he was under severe hypoxemia. We suspected miliary tuberculosis with acute respiratory failure due to his clinical course. The intensive therapy with antituberculosis drugs, hydrocortisone (1 g/day) was started and intubation was performed. The smear of the clinical samples did not show acid-fast bacilli on admission. However, by the polymerase chain reaction amplification method specific DNA fragments of Mycobacterium tuberculosis complex were detected from BALF and bone marrow. He was extubated after the 13th day and he recovered. It is difficult to separate and identify tubercle bacilli rapidly, but PCR is available for early diagnosis.

Acute Disease↗
PubMed1994

NADH dehydrogenase subunit genes in the mitochondrial DNA of yeasts.

The genes encoding the NADH dehydrogenase subunits of respiratory complex I have not been identified so far in the mitochondrial DNA (mtDNA) of yeasts. In the linear mtDNA of Candida parapsilosis, we found six new open reading frames whose sequences were unambiguously homologous to those of the genes known to code for NADH dehydrogenase subunit proteins of different organisms, i.e., ND1, ND2, ND3, ND4L, ND5, and ND6. The gene for ND4 also appears to be present, as judged from hybridization experiments with a Podospora gene probe. Specific transcripts from these open reading frames (ND genes) could be detected in the mitochondria. Hybridization experiments using C. parapsilosis genes as probes suggested that ND genes are present in the mtDNAs of a wide range of yeast species including Candida catenulata, Pichia guilliermondii, Clavispora lusitaniae, Debaryomyces hansenii, Hansenula polymorpha, and others.

Amino Acid Sequence↗
PubMed1994

[Pulmonary nocardiosis concurrent with Pneumocystis carinii pneumonia in a case of smoldering adult T-cell leukemia].

A 76-year-old man, with concomitant pulmonary nocardiosis and Pneumocystis carinii (PC) pneumonia, is described. He suffered from smoldering type adult T-cell leukemia and strongyloidiasis. Chest X-rays showed bilateral infiltrative shadows. Nocardia asteroides was isolated from sputum and bronchofiberscopic aspirates. PC was not detected, however, despite the chest X-ray raising the suspicion of PC pneumonia. Since PC antigen was detected by immunoelectrophoresis in the absence of other clinical or histologic data, we diagnosed a latent PC infection. He improved with the use of cefuzonam and ST compounds, but experienced recurrence five months after discharge. N. asteroides and PC were detected from sputum on the second admission. As multiple concurrent infections with opportunistic organisms are not uncommon in immunocompromised hosts, we must maintain a high index of suspicion for opportunistic infection in cases such as this one.

Aged↗
PubMed1993

DNA sequence analysis of a 17 kb fragment of yeast chromosome XI physically localizes the MRB1 gene and reveals eight new open reading frames, including a homologue of the KIN1/KIN2 and SNF1 protein kinases.

We report in this paper the sequence of a part of chromosome XI of Saccharomyces cerevisiae. This 17 kbp nucleotide sequence represents the right half of cosmid pUKG151 and contains nine open reading frames, YKL453, 450, 449, 448, 445, 443, 442, 441 and the 5' part of YKL440. YKL440 was previously identified as the MBR1 gene and plays a role in mitochondrial biogenesis. YKL443 is a homologue of the yeast serine-rich protein (SRP1), while YKL453 presents strong homologies with the KIN1/KIN2/SNF1 kinase family. It must be pointed out that the size of this gene is well above average for yeast.

Amino Acid Sequence↗
PubMed1993

The DNA sequence analysis of the HAP4-LAP4 region on chromosome XI of Saccharomyces cerevisiae suggests the presence of a second aspartate aminotransferase gene in yeast.

The nucleotide sequence of a 19,000 base pair region from the left arm of chromosome XI of Saccharomyces cerevisiae has been determined and analysed. It covers the HAP4-GFA1-LAP4 loci already described. As expected HAP4, GFA1 and LAP4 genes have been found and six new open reading frames (ORFs) with a coding capacity of more than 100 amino acid residues have been identified. One of them (YKL461) shows a high degree of identity with an aspartate aminotransferase gene. This raises the question of a second aspartate aminotransferase gene in yeast. A second ORF (YKL462) shows features compatible with a membranous localization. The other ORFs do not show a similarity with any known gene. A member of the highly repetitive 'CAT' DNA sequence is present.

Aspartate Aminotransferases↗