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Biomedical subjects

K Yamane

Publications and source records attributed to K Yamane.

At least 145 records · Page 8Linked to original sources

The 25 degrees-36 degrees region of the Bacillus subtilis chromosome: determination of the sequence of a 146 kb segment and identification of 113 genes.

We determined a 146 kb contiguous sequence at the 25 degrees-36 degrees region of the Bacillus subtilis chromosome containing the amyE-srfA segment. Among the 113 ORFs identified, 33 are already known. functions were assigned to 38 ORFs by a search of non-redundant protein sequence data banks and those of 16 ORFs were suggested through significant similarity with reported sequences. The amino acid sequences of 13 of the ORfs were similar to proteins of unknown function of Escherichia coli, Haemophilus influenzae and other species. We did not find similarities for 29 ORFs to any known proteins. The 146 kb region is rich in enzymes (35 ORFs) related to the metabolism of low molecular mass compounds and five genes for surfactin production occupy about 26 kb of the region.

Bacillus subtilis↗

A Bacillus subtilis secreted phosphodiesterase/alkaline phosphatase is the product of a Pho regulon gene, phoD.

A secreted phosphodiesterase/alkaline phosphatase, APaseD, was purified from a culture of Bacillus subtilis JH646MS. Its phosphodiesterase activity was reminiscent of an APase isolated and characterized previously. Immunoassay and N-terminal sequencing showed the two proteins to be identical. Using the first 20 amino acids of the mature protein, a BLAST search of GenBank was used to find an homologous sequence. An exact match was found but in a putative non-coding region. It was hypothesized that there was a base pair deletion in the phoD gene. A DNA fragment internal to the coding region was generated by PCR using template DNA from a strain which produced APaseD. The PCR fragment was cloned and used to interrupt the gene. Western blot analysis of the parent and the mutated strains showed that APaseD was missing in the mutant. Resequencing of the gene revealed a larger ORF encoding a protein similar in size to the 49 kDa APaseD estimated by SDS-PAGE. The promoter was then cloned, sequenced and used in phoD-lacZ promoter fusions which showed that the gene was phosphate-starvation-induced and dependent on PhoP and PhoR for expression.

Alkaline Phosphatase↗

Oxygen-controlled regulation of the flavohemoglobin gene in Bacillus subtilis.

A gene, hmp, which encodes a ubiquitous protein homologous to hemoglobin was isolated among genes from Bacillus subtilis that are induced under anaerobic conditions. The hmp protein belongs to the family of two-domain flavohemoproteins, homologs of which have been isolated from various organisms such as Escherichia coli, Alcaligenes eutrophus, and Saccharomyces cerevisiae. These proteins consist of an amino-terminal hemoglobin domain and a carboxy-terminal redox active site domain with potential binding sites for NAD(P)H and flavin adenine dinucleotide. The expression of hmp is strongly induced upon oxygen limitation, and the induction is dependent on a two-component regulatory pair, ResD and ResE, an anaerobic regulator, FNR, and respiratory nitrate reductase, NarGHJI. The requirement of FNR and NarGHJI for hmp expression is completely bypassed by the addition of nitrite in the culture medium, indicating that fnr is required for transcriptional activation of narGHJI, which produces nitrite, leading to induction of hmp expression. In contrast, induction of hmp was still dependent on resDE in the presence of nitrite. A defect in hmp in B. subtilis has no significant effect on anaerobic growth.

Anaerobiosis↗

Predicting infection in hospitalized patients with systemic lupus erythematosus.

To identify the risk factors for infection and to develop a model for the prediction of infection in hospitalized patients with active systemic lupus erythematosus (SLE), we carried out a retrospective cohort study with clinical data collected from 121 consecutive patients with active SLE. Seventeen patients had infection within 6 months of beginning steroid therapy for active SLE. Independent multivariate predictors of infection were a decrease in the serum albumin value, an increase in the serum creatinine value, and prednisolone use in a dose of > or = 60 mg/day without methylprednisolone pulse therapy. The error rate of the model by 10-fold cross-validation method was 12%, sensitivity was 65%, specificity was 91%, and positive predictive value was 55%. Four nonsurvivors were correctly discriminated. Use of this model could contribute to earlier diagnosis of infection and may assist decisions regarding empiric antimicrobial administration in patients with SLE.

Adolescent↗

[A dual occurrence of endometrial and gastric adenocarcinomas in systemic lupus erythematosus].

The association of systemic lupus erythematosus (SLE) and solid tumors is infrequent. Furthermore, there has been no previous description of multiple occurrence of carcinoma in a single patient with SLE. We describe clinical picture of the patient who had dual occurrence of endometrial and gastric adenocarcinomas in addition to clinically active SLE and organ system involvement.

Adenocarcinoma↗

[Clinical study of pneumoconiosis featuring small nodules and without large opacities on high-resolution CT].

High-resolution CT (HRCT) was performed in 25 patients with pneumoconiosis diagnosed as category 1 or 2 according to the ILO radiographic classification. The small opacities shown on HRCT images were classified as small branching structures (SBS) or small rounded opacities (SRO). Patients in these two groups were compared with respect to the distribution of small nodules, pulmonary function, arterial blood gas analysis, degree of exertional dyspnea, and alteration ratio of the lung field CT numbers between maximum inspiration and maximum expiration (%(I-E)). The small nodules in all patients of the SBS group were classified as type p. SBS were distributed evenly throughout the lungs, while SRO were mainly localized in the upper middle lobe. Pulmonary emphysema was observed in patients with SBS and was particularly common among patients in this group with an irregular septum (IS). No emphysematous lesions were detected in the SRO group. RV/ TLC was higher and exertional dyspnea was more severe in the SBS group than in the SRO group. It was considered that an increase of RV/TLC resulted in worsening of exertional dyspnea in the patients with SBS. The % (I-E) value was significantly lower in the SBS group than in the SRO group.

Aged↗

[Usefulness of reaction times in high-cortical recognition tasks].

We compared reaction times for high-cortical recognition tasks with kana-searching test in a group of 46 normal individuals aged 21 to 83 who gain a full score in Mini-Mental State. The following tasks of reaction time were used; simple reaction time (SRT), choice reaction time (CRT), memory scanning time (MST), and visual searching time (VST). The results were consistent and demonstrated that mean reaction time of 46 individuals increased in difficult tasks such as MST or VST. All reaction times increased depending on aging, which became longer in difficult tasks. Score of kana-searching test was correlated with MST and VST, but did not correlate with SRT and CRT. These findings suggest that reaction times may be useful for mass-screening of high-cortical functions in different ages as well as the kana-searching test.

Adult↗

Binding affinity and particle size of LDL in subjects with moderate hypercholesterolemia: relationship with in vivo LDL metabolism.

The aim of this study was to examine relationships between low density lipoprotein (LDL) metabolism, in vitro binding, and particle size. Twenty four study subjects, 17 men and 7 women, had elevated plasma total cholesterol (TC), ranging from 174 to 232 mg/dl, and LDL cholesterol (LDLC) ranging from 113 to 195 mg/dl after 12 weeks on a Step I diet. The fractional clearance rate (FCR) for LDL ranged from 0.233 to 0.619 pools/day (0.366 +/- 0.021) and was significantly correlated with plasma triglycerides (TG) (P < 0.05). Although there was no relation between FCR and binding in the study group as a whole, those subjects with an FCR within the normal range (< 0.45, n = 20), showed a significant negative correlation between FCR and the KD for LDL binding, (r = 0.52). A subset of four subjects with an elevated FCR (> 0.45) had higher production rate (PR) (P < 0.005) and a significant positive correlation between the KD for LDL binding and FCR (P < 0.05). LDL size varied from 240.0 to 265.8 A and was significantly inversely correlated with plasma TG (P < 0.001) but there was no relation between LDL size and metabolism or binding affinity. Thus, there appears to be a correlation between binding affinity and clearance for subjects who had normal LDL production and clearance rates. On the other hand, mechanisms other than binding affinity appear to influence clearance in subjects with elevated rates of production and clearance.

Adult↗

Depletion of small cytoplasmic RNA confers fusidic-acid resistance on Bacillus subtilis.

Bacillus subtilis small cytoplasmic RNA (scRNA) is a member of a signal recognition particle (SRP)-like RNA family. To analyze the function of scRNA in protein synthesis, a B. subtilis strain SC201NA was constructed in which the expression of intact scRNA is regulated by an IPTG-inducible promoter. In this strain, depletion of scRNA leads to deficient translation and sporulation as well as morphological changes. In addition, the growth of SC201NA in the absence of IPTG became fusidic-acid resistant. The acquisition of fusidic-acid resistant phenotype by depletion of scRNA suggested that scRNA is associated with elongation factor G (EF-G) in the translation process.

Bacillus subtilis↗

In vitro regulation of low-density lipoprotein receptor interaction by fatty acids.

Low-density lipoprotein (LDL) receptor binding is the initial step in receptor-mediated clearance. Dietary fat composition is known to affect LDL clearance, but the mechanism of the effect is unknown. We have examined the effects of altered membrane fatty acid composition, as might occur when specific dietary fats are consumed, on LDL binding using a Chinese hamster ovary (CHO) line that constitutively expresses the human LDL receptor. Binding of pooled human LDL to its receptor was compared in cells enriched with various fatty acids. Binding affinity was greater (lower Kd) for cells grown in 16:0-, 18:0-, or 18:1-enriched media than for those grown in 18:2 (P < .0001). The apparent receptor number (Bmax) was lower for cells enriched in saturated fatty acids and 18:1. Fluidity was assessed by measuring diphenylhexatriene (DPH) fluorescence anisotropy (rs). Cells enriched in 18:1 or 18:2 were the most fluid (P < .003). The correlation between binding and fluidity (r = .24, P = .27) was weak and did not appear to explain the effects of fatty acid modification on LDL receptor binding. Thus, it appears that cellular enrichment in 16:0, 18:0, and 18:1 increases binding affinity by affecting properties other than membrane fluidity. Changes in Bmax may also contribute to the observed differences in LDL binding.

Animals↗

Clinical findings in patients with recurrent intracerebral hemorrhage.

BACKGROUND: The use of computerized tomography has led to the detection of second intracerebral hemorrhage (ICH) in some patients. There have, however, only been a few clinical studies of second ICH. SUBJECTS AND METHODS: Thirty patients with a second ICH were analyzed according to clinical criteria. These patients comprised 5.9% of all patients admitted to Chugoku Rosai Hospital for ICH between 1984 and 1992. RESULTS: The mean interval between the first and second ICH was 27.7 months (range 1-144). The incidence of second hemorrhage was especially high within the first year after the initial ICH. Twelve patients bled bilaterally into the basal ganglia or thalamus during either the first or second attack. Most of these patients had poor outcomes and prognoses. The nine patients with good prognoses included patients with high activity of daily living (ADL) prior to the second attack, and those with neurologic grade 1 following the second attack. All of these patients were managed with conservative therapy. The 10 patients who underwent surgery had poor prognoses. CONCLUSIONS: The incidence of second ICHs is greater than that of initial ICHs. Many patients who experience a second ICH will have a poor prognosis, possibly worse than expected. Surgical therapy should not be recommended in such patients, since our data suggest that they will not do well even after surgery.

Activities of Daily Living↗

Computer simulation of tablet motion in coating drum.

PURPOSE: The purpose is to develop a new method of finding optimal conditions in tablet coating operations with the use of computer simulation. METHODS: DEM(Discrete Element Method), which calculates trajectories of individual tablets by numerically integrating Newton's equations of motion, was used in this simulation. RESULTS: The effects of tablet size, tablet loading, rotation speed on mean surface time, circulation time, and surface time deviation agree qualitatively with experimental results by other workers. CONCLUSIONS: It is found that DEM is a useful method of predicting tablet motion in a coating drum.

Computer Simulation↗

srb: a Bacillus subtilis gene encoding a homologue of the alpha-subunit of the mammalian signal recognition particle receptor.

We cloned a Bacillus subtilis gene (srb) encoding a homologue of the mammalian signal recognition particle receptor alpha-subunit (SR alpha). The gene is 987 bp in length and encodes a 329-amino acid protein. The deduced amino acid sequence of the protein shared 26.6, 36.2 and 49.7% identity with those of mammalian SR alpha, archaebacterial DP alpha and Escherichia coli FtsY, respectively. The protein contains three conserved GTP-binding elements like the other three SRP receptor proteins, though the N-terminal portion of the putative B. subtilis protein was shorter than the others. Secondary structure prediction showed than an amphipathic alpha-helix is positioned in the N-terminal region. A defect in srb inhibited cell growth and protein translocation.

Amino Acid Sequence↗

Acquisition of azide-resistance by elevated SecA ATPase activity confers azide-resistance upon cell growth and protein translocation in Bacillus subtilis.

We isolated four azide-resistant secA mutants of Bacillus subtilis and found that all of them were the result of a single amino acid replacement of threonine 128 of SecA by alanine or isoleucine. In the presence of 1.5 mM sodium azide, cell growth and protein translocation of the wild-type strain were completely inhibited, but those of the azide-resistant mutant strains were not. Wild-type and two mutant SecA proteins were purified. Both the basal level and the elevated ATPase activity of the mutant SecA proteins were threefold higher than those of the wild-type SecA. The elevated ATPase activity of the SecA mutants was reduced upon the addition of 1.5 mM sodium azide by only 5-10% as compared with 40% for that of the wild-type. These results indicate that the elevated ATPase activity of the SecA mutants is resistant to sodium azide and that is also required for the protein translocation process of B. subtilis.

Adenosine Triphosphatases↗

Small cytoplasmic RNA (scRNA) gene from Clostridium perfringens can replace the gene for the Bacillus subtilis scRNA in both growth and sporulation.

Small cytoplasmic RNA (scRNA) is a member of an evolutionarily conserved signal-recognition-particle-like RNA family. Using a DNA fragment of Bacillus subtilis scRNA gene as a probe, we cloned and characterized a Clostridium perfringens gene encoding the scRNA. Mapping the 5' and 3' ends of scRNA revealed that C. perfringens scRNA consists of 269 nucleotides: the sequence has about 70% primary sequence homology with B. subtilis scRNA. The predicted secondary structure appeared to be similar to that of B. subtilis scRNA, indicating that there are domains I and II in C. perfringens scRNA, in addition to domain IV. Functional analysis showed that C. perfringens scRNA could compensate for vegetative growth and allow the formation of heat-resistant spores in an scRNA-depleted B. subtilis strain, whereas Escherichia coli 4.5S RNA could not maintain sporulation. Since both E. coli 4.5S RNA and C. perfringens scRNA have the same binding specificity to B. subtilis Ffh protein, the difference in complementation activity reflects the function of domains I and II.

Amino Acid Sequence↗

Determination of a 17,484 bp nucleotide sequence around the 39 degrees region of the Bacillus subtilis chromosome and similarity analysis of the products of putative ORFs.

We have determined a 17,484 bp nucleotide sequence around the 39 degrees region, located about 480 kb downstream from the zero position of the Bacillus subtilis chromosome physical map. Among the 17 putative ORFs identified, orf1 and orf2 seem to correspond to mtlA and mtlB, encoding mannitol-specific phosphotransferase enzyme II and mannitol-1-phosphate dehydrogenase, respectively. orf4 seems to be another signal peptidase I gene (sipS) of B. subtilis. The putative products of six ORFs were similar to known proteins in data banks, namely a hypothetical 29.7 kDa protein of Escherichia coli (Orf7), a lactam utilization protein (Orf8), the urea amidolyase of yeast (Orf12), the IcIR regulatory protein for aceAB of Salmonella typhimurium (Orf13), penicillin-binding protein 2 (Orf16) and aryl-alcohol dehydrogenase (Orf17). The amino acid sequence of Orf3 showed 34% identity with that of LeuC of B. subtilis, though they seem to be functionally different. The remaining seven ORFs did not show similarity to any known proteins.

Amino Acid Sequence↗