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Biomedical subjects

K S Bae

Publications and source records attributed to K S Bae.

35 records · Page 2Linked to original sources

Kitasatospora cheerisanensis sp. nov., a new species of the genus Kitasatospora that produces an antifungal agent.

An actinomycete, strain YC75T, which produced bafilomycin-like antifungal compounds, was identified as a member of the genus Kitasatospora on the basis of morphological and chemotaxonomic characteristics. The strain produced the aerial and fragmenting vegetative mycelia consisting of straight chains of 20 or more smooth-surfaced spores. Submerged spores were formed in tryptic soy broth. No soluble pigments were formed. Whole-cell hydrolysates contained glucose and mannose, but not galactose. The 16S rDNA sequence of YC75T was compared with those of the other representative kitasatosporae and streptomycetes. Strain YC75T formed a significant monophyletic clade with Kitasatospora phosalacinea. The levels of DNA relatedness between strain YC75T and representatives of the genus Kitasatospora ranged from 16 to 59% including K. phosalacinea (28 and 40%). It is clear from polyphasic evidence that the isolate should be classified as Kitasatospora cheerisanensis sp. nov., whose type strain is YC75T (= KCTC 2395T). The presence of galactose in whole-cell hydrolysates may not be a stable chemical marker for the genus Kitasatospora.

Antifungal Agents↗

Amycolatopsis thermoflava sp. nov., a novel soil actinomycete from Hainan Island, China.

A soil isolate, which had been assigned to the genus Nocardia, was shown to have properties consistent with its classification in the genus Amycolatopsis. An almost complete nucleotide sequence of the 16S rDNA of the strain was determined following cloning and sequencing of the amplified gene. The sequence was aligned with those available for members of the family Pseudonocardiaceae and related taxa and phylogenetic trees were inferred using three tree-making algorithms. The organism consistently formed a distinct monophyletic clade with the type strain of Amycolatopsis methanolica, but DNA-DNA relatedness data showed that the two strains belonged to distinct genomic species. The organism was also distinguished from the type strains of all validly described species of Amycolatopsis using a battery of phenotypic properties. The genotypic and phenotypic data show that the strain merits recognition as a new species of the genus Amycolatopsis. The name proposed for the new species is Amycolatopsis thermoflava sp. nov. The type strain is IFO 14333T.

Actinomycetales↗

Cloning and phylogenetic analysis of chitin synthase genes from the insect pathogenic fungus, Metarhizium anisopliae var. anisopliae.

Degenerated PCR primers were used to amplify chitin synthase genes from genomic DNA of Metarhizium anisopliae var. anisopliae. Through cloning and sequencing of approximately 600-bp fragments amplified by PCR, we found three genes encoding different types of chitin synthases, designated MaCHS1, MaCHS2, and MaCHS3. Southern blot analysis performed on genomic DNA showed that each of the chitin synthases MaCHS1, MaCHS2, and MaCHS3 is encoded by a single copy gene. Alignment of their deduced amino acid sequences with those of other euascomycetes separated the sequences into three distinct classes. MaCHS1 was identified as a gene for class I chitin synthase, MaCHS2 for class II, and MaCHS3 for class III. The UPGMA dendrogram and phylogenetic tree of the deduced amino acid sequences revealed the taxonomic and evolutionary position of Metarhizium anisopliae var. anisopliae.

Amino Acid Sequence↗

Development of an anaerobic bacterial leakage model.

The majority of bacteria associated with infections of endodontic origin are strict anaerobes. The purpose of this study was to develop an endodontic microleakage model using strict anaerobic bacteria in a two-chamber system. Nine species of anaerobic bacteria were tested for viability and detection by either turbidity or color change of the broth. A survey of pH chromogenic substrates revealed that bromcresol purple (pH 5.2 = yellow, pH 6.8 = purple) could be used as a chromogenic indicator to detect the growth of anaerobic bacteria. Peptone-yeast extract-glucose broth (PYG) and brain heart infusion broth (BHI) were each used alone and with bromcresol purple (bpPYG, bpBHI) in this study. Fusobacterium nucleatum and F. necrophorum were viable in all four media for > 2 wk and produced both turbidity and a color change after only 1 day of incubation. Veillonella parvula in either bpBHI or BHI and Peptostreptococcus anaerobius in either bpPYG or BHI were viable for > 2 wk and showed a color change or turbidity after 1 or 2 days. The results indicate that leakage of strict anaerobes may be evaluated in a two-chamber system.

Actinomyces↗

Black-pigmented bacteria in coronal and apical segments of infected root canals.

Black-pigmented bacteria (BPB) have been associated with infections of endodontic origin. The purpose of this study was to culture and identify BPB from the apical and coronal segments of infected root canals to understand better their ecological relationships. Teeth with a periapical radiolucency were extracted and immediately placed in reduced transport fluid for transport to an anaerobic chamber. Of 18 sampled roots, 12 were positive for the growth of BPB. Eight of the 12 roots with BPB had a carious exposure of the pulp chamber. Seven roots had Prevotella nigrescens in both the apical and the coronal segments. Six of these seven teeth had carious exposures of the pulp chamber. Of the 12 roots infected with BPB, six roots had two different species of BPB, with P. nigrescens always being one of the species. P. nigrescens was the most often isolated BPB from both the coronal and apical segments of infected root canals.

Colony Count, Microbial↗

Perforation repair comparing mineral trioxide aggregate and amalgam using an anaerobic bacterial leakage model.

The purpose of this study was to evaluate the ability of mineral trioxide aggregate (MTA) and amalgam to seal furcal perforations in extracted human molars using an anaerobic bacterial leakage model. Furcal perforations were made in 39 maxillary and mandibular human molars with a high-speed bur. These were randomly divided into two experimental groups of 18, with the remaining three teeth used as positive controls. Experimental group 1 was repaired with MTA and group 2 with amalgam. Three additional teeth without perforations served as negative controls. A dual chamber anaerobic bacterial leakage model was assembled. Brain heart infusion broth with yeast extract, hemin, menadione, and the chromogenic indicator bromcresol purple was used as the culture broth for Fusobacterium nucleatum. Eight of 18 amalgam samples leaked, whereas none of the 18 MTA samples leaked. MTA was significantly better than amalgam in preventing leakage of F. nucleatum past furcal perforation repairs.

Aluminum Compounds↗

Molecular cloning of chitinase cDNAs from the silkworm, Bombyx mori and the fall webworm, Hyphantria cunea.

cDNAs encoding chitinases were cloned and characterized from Bombyx mori and Hyphantria cunea, and their gene expression during the metamorphosis was also studied. The chitinase cDNA from B. mori encodes a protein of 565 amino acids with a calculated molecular mass of 63.4 kDa and the H. cunea chitinase cDNA encodes a protein of 553 amino acids with a calculated molecular mass of 62.0 kDa. Amino acid alignment of the two chitinases revealed 75% homology and 77-80% with M. sexta chitinase. The putative cleavage site of the signal peptide was between amino acid residues 20 and 21 for both chitinases. There were three potential N-glycosylation sites in the chitinase of B. mori at the amino acid residues 86-89, NFTS 304-307, NATG, 398-401, NYTV, whereas two potential N-glycosylation sites were present at the amino acid residues 86-89, NFTA and 304-307, NATG, in that of H. cunea. Southern blot analysis of total genomic DNA suggested that the B. mori genome has only one chitinase gene detectable by the cDNA probe and the H. cunea genome has one or two chitinase gene copies. Northern analysis indicated that gene expression was up-regulated during the molting process, larval-pupal transformation and pupal-adult transformation, when enzymatic degradation of cuticle was occurring.

Amino Acid Sequence↗

Nocardia flavorosea sp. nov.

An actinomycete strain, 'Nocardai flavorosea' JCM 3332, was found to have properties consistent with its classification in the genus Nocardia. An almost complete gene sequence of the 16S rDNA of the strain was determined following cloning and sequencing of the amplified gene. The sequence was aligned with those available for nocardiae and phylogenetic trees were inferred using four tree-making algorithms. The organisms consistently formed a distinct clade with the type strain of Nocardia carnea. However, DNA relatedness experiments showed that the strain and N. carnea DSM 43397T belonged to two distinct genomic species. The organism was also distinguished from representative of all of the validly described species of Nocardia using a combination of phenotypic properties. These genotypic and phenotypic data show that the strain merits recognition as a new species of the genus Nocardia. The name proposed for the new species of is Nocardia flavorosea sp. nov. The type strain is JCM 3332T.

Base Sequence↗

Occurrence of Prevotella nigrescens and Prevotella intermedia in infections of endodontic origin.

The occurrence of Prevotella intermedia and Prevotella nigrescens in endodontic infections was studied using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of whole cell protein to distinguish between the species. Previous studies have shown an association between black-pigmented bacteria (BPB) and endodontic infections and that Prevotella intermedia (previously known as Bacteroides intermedius) was the most commonly isolated BPB. Recently, however, strains identified as P. intermedia were shown to in fact be composed of two separate species, P. intermedia and P. nigrescens. Fifty-six strains of BPB isolated from endodontic infections and previously identified as P. intermedia were used in this study. Following SDS-PAGE, P. nigrescens showed a unique 18.6 kDa band that was used to differentiate P. nigrescens from P. intermedia. Of the 56 strains of BPB, 41 (73.2%) were identified as P. nigrescens and 15 (26.8%) as P. intermedia. This study confirms that P. nigrescens, and not P. intermedia, is the BPB most often isolated from infections of endodontic origin.

Amino Acid Sequence↗

MR566A and MR566B, new melanin synthesis inhibitors produced by Trichoderma harzianum. I. Taxonomy, fermentation, isolation and biological activities.

New melanin synthesis inhibitors (MR566A and B) and six related known isocyanocyclopentenes were isolated from the fermentation broth of Trichoderma harzianum. The IC50 values of MR566A and B against mushroom tyrosinase were 1.72 and 47 microM, respectively. They inhibited melanin biosynthesis in B16 melanoma cells with MIC values of 0.1 and 2.2 microM, respectively. Also isolated from the same culture extract of T. harzianum was a new oxazole (MR93B), which showed no inhibitory activity against mushroom tyrosinase at a concentration of 1,000 microg/ml.

Animals↗

Cloning and characterization of chitin synthase gene fragments from Penicillium chrysogenum.

DNA fragments homologous to chitin synthase were amplified from the genomic DNA of Penicillium chrysogenum by PCR. Cloning and sequencing of the PCR-amplified fragments led to the identification of four different genes, designated PcCHS1, PcCHS2, PcCHS3, and PcCHS4. By comparison of the deduced amino acid sequences, PcCHS1 was identified as a gene for class I chitin synthase, PcCHS2 and PcCHS3 were for class II, and PcCHS4 was for class III. Among these only PcCHS4 includes an intervening sequence of 56 bp. The analysis of the deduced amino acid sequences revealed a close evolutionary relationship between Penicillium and ascomycetous fungi.

Amino Acid Sequence↗

How important is the role of the internal anal sphincter in fecal continence? An experimental study in dogs.

It is a generalized concept that the internal anal sphincter (IAS) plays a significant role in fecal continence by generating high pressure in the anal canal at rest and relaxation during rectal distention. Agreement also exists on the importance of internal sphincter-saving anoplasty on anorectal malformations in establishing anal continence. Twelve dogs were divided into four groups. Group 1, a control group, was subjected to a perirectal dissection only. Group 2 underwent the same perirectal dissection plus a 2-cm resection of the anal canal. Group 3 underwent the perirectal dissection plus a 4-cm resection of the anal canal, and group 4 underwent perirectal dissection and transposition of the anus to the posterolateral portion of the voluntary muscle mass. Clinical continence was evaluated, and manometric results were compared with preoperative measurements. All dogs in groups 1, 2, and 3 were clinically continent without soiling except one in group 3, and also manometric results showed minimal change between preoperative and postoperative anal pressure profiles. The transposed anus of group 4 showed continuous fecal soiling. The anal resting pressure (ARP) was also decreased but still existed in this group. This experimental study showed that the IAS contributes to the anal resting tone. However, resection of the IAS did not completely interfere with fecal continence. The smooth muscle of pulled-through rectum seemed to partly take over the function of the IAS.

Anal Canal↗

Detection of laryngeal function using speech and electroglottographic data.

The purpose of this research was to develop quantitative measures for the assessment of laryngeal function using speech and electroglottographic (EGG) data. We developed two procedures for the detection of laryngeal pathology: 1) a spectral distortion measure using pitch synchronous and asynchronous methods with linear predictive coding (LPC) vectors and vector quantization (VQ) and 2) analysis of the EGG signal using time interval and amplitude difference measures. The VQ procedure was conjectured to offer the possibility of circumventing the need to estimate the glottal volume velocity wave-form by inverse filtering techniques. The EGG procedure was to evaluate data that was "nearly" a direct measure of vocal fold vibratory motion and thus was conjectured to offer the potential for providing an excellent assessment of laryngeal function. A threshold based procedure gave 75.9 and 69.0% probability of pathological detection using procedures 1) and 2), respectively, for 29 patients with pathological voices and 52 normal subjects. The false alarm probability was 9.6% for the normal subjects.

Adult↗

Cloning of the human erythropoietin receptor gene.

We have isolated and characterized a genomic clone of the human erythropoietin (Epo) receptor from a placental genomic library using a cDNA probe for the murine Epo receptor. The coding region spans about 6.5 kb with seven intervening sequences ranging in size from 81 bp to 2.1 kb. A stretch of 123 purines is found in the 5' region from -456 to -578 upstream from the first codon and flanking the Alu repetitive sequences located further upstream. The human Epo receptor contains a palindromic sequence 5' of the translated region that consists of an almost perfect inverted repeat of 12 nucleotides (CAGCTGC(G/C)TCCG) centered about G at -92 from the first codon. An inverted SP1 binding site (CCGCCC) and an inverted GATA-1 binding site (TTATCT) are located at positions -151 and -179, respectively, and CACCC sequences are located at -585 and further upstream. No TATA or CAAT sequences are in this 5' flanking region. However, this region as far as -275 has a 72% GC content compared with an overall GC content of 56%. A 1-kb BamHI fragment of the human Epo receptor containing 700 bp of sequences 5' of the coding region was transcribed in an in vitro transcription assay; initiation of transcription appeared to be around 132 +/- 5 just downstream from the inverted SP1 site at -151. T1 analysis of human Epo receptor messenger RNA also maps the site of transcription initiation to this region. Within 180 nucleotides 5' to the first exon are three regions with 70% or greater homology with the murine Epo receptor. The study of this gene, including its similarities with the murine Epo receptor, should help elucidate aspects of the transcriptional and possible translational control of the Epo receptor in human erythroid cells and thus its role in signal transduction and erythroid differentiation.

Amino Acid Sequence↗

Alkaline Phosphatase and Other Hydrolyases Produced by Cenococcum graniforme, an Ectomycorrhizal Fungus.

Cell extracts of Cenococcum graniforme have been found to contain the following hydrolytic enzymes: protease, esterase, alpha-d-galactopyranosidase, beta-d-galactopyranosidase, alpha-d-mannopyranosidase, beta-d-xylopyranosidase, alpha-d-glucopyranosidase, beta-d-glucopyranosidase, and alkaline phosphatase. Sulfatase, inorganic pyrophosphatase, and beta-d-mannopyranosidase were not detected in the extracts. beta-d-Xylopyranosidase and alpha-d-mannopyranosidase were most active in the neutral pH range, protease and phosphatase were most active in the alkaline pH range, and other enzymes were most active in the acidic pH range. These enzymes showed a high association with cell wall material, and the release of enzymes from the cells into the culture fluid appeared to occur only when the cells were undergoing autolysis. Alkaline phosphatase in C. graniforme is a constitutive enzyme, and examination of the alkaline phosphatase following a purification of 265-fold produced the following characteristics: pH optimum of 9.5, M(r) of 60,000, K(m) of 2.1 x 10 M for p-nitrophenylphosphate, and activation energy for hydrolysis of the substrate at 9.9 kcal (1 cal = 4.184 J)/mol.

Journal Article↗