Pseudomonas insulae sp. nov., isolated from island soil.

A novel Gram-stain-negative, rod-shaped, aerobic and motile bacterium designated strain UL073T was isolated from a forest soil of an island, and subjected to taxonomic characterization. Strain UL073T grew at 10-37 °C (optimum, 30 °C), at pH 5.0-10.0 (optimum, pH 7.0) and in the presence of 0-3 % NaCl (optimum, 0 %), respectively. Strain UL073T showed the highest sequence similarity to Pseudomonas lalkuanensis PE08T based on 16S rRNA gene analysis with a sequence similarity of 98.08 %, which was well below the suggested cutoff for species distinction. The 16S rRNA gene tree as well as the multilocus sequence analysis and genome-based trees indicated the independent taxonomic position of strain UL073T, and the orthologous average nucleotide identity and in silico DNA-DNA hybridization values between strain UL073T and related species were no higher than 84.7 and 28.3% respectively, thus confirming the distinctive taxonomic position of the strain. The chemotaxonomic properties were consistent with those of the genus, as the major fatty acids of the strain were a summed feature consisting of C18 : 1 ω7c/C18 : 1 ω6c (31.4 %), another summed feature consisting of C16 : 1 ω7c/C16 : 1 ω6c (23.1 %), and C16 : 0 (22.0 %), the major respiratory quinone was ubiquinone 9, and the major polar lipids were phosphatidylethanolamine and diphosphatidylglycerol. The genome size and DNA G+C content of strain UL073T were 4.87 Mbp and 65.9 mol%. On the basis of phenotypic and phylogenetic evidence, strain UL073T should be classified as representing a novel species of Pseudomonas, for which the name Pseudomonas insulae sp. nov. (type strain=UL073T=KCTC 82407T=JCM 34511T) is proposed.

Pseudomonas guryensis sp. nov. and Pseudomonas ullengensis sp. nov., isolated from soil.

Two Gram-staining-negative, aerobic, rod-shaped bacteria designated strains SR9T and UL070T, were isolated from soil and subjected to taxonomic characterization. Strain SR9T grew at 10-37 °C (optimum 30 °C), at pH 4.0-10.0 (optimum pH 8.0) and in the presence of 0-1 % NaCl (optimum 0 %), and UL070T at 4-33 °C (optimum 30 °C), at pH 4.0-10.0 (optimum pH 7.0) and in the presence of 0-2 % NaCl (optimum 0 %), respectively. Strain UL070T was motile with flagella. Analysis of 16S rRNA gene sequences indicated that the two strains fell into phylogenetic clusters belonging to the genus Pseudomonas. Both strains SR9T and UL070T were mostly related to Pseudomonas campi S1-A32-2T with 99.70 and 99.01% sequence similarities, and the similarity between the two isolates was 98.90 %. The genome-based in silico analyses indicated that each of the strains SR9T and UL070T was clearly separated from other species of Pseudomonas, as the orthologous average nucleotide identity (OrthoANI) and the digital DNA-DNA hybridization (dDDH) values were no higher than 93.09 and 50.03% respectively with any related species, which were clearly below the cutoff for species distinction. The fatty acid profiles of the two strains mainly consisting of unsaturated components, the presence of ubiquinone 9 (Q-9) as the major respiratory quinone, and phosphatidylethanolamine (PE) and diphosphatidylglycerol (DPG) as the diagnostic polar lipids were consistent with their classification into Pseudomonas. The DNA G+C contents of strains SR9T and UL070T were 63.2 mol% and 63.6 mol% respectively. On the basis of both phenotypic and phylogenetic evidences, each of the isolated strains should be classified as a novel species, for which the names Pseudomonas guryensis sp. nov. (type strain=SR9T=KCTC 82228T=JCM 34509T) and Pseudomonas ullengensis sp. nov. (type strain=UL070T=KCTC 82229T=JCM 34510T) are proposed.

The protease inhibitor, elafin, induces p53-dependent apoptosis in human melanoma cells.

Expression of the protease inhibitor elafin is deregulated in several human cancers. However, functions of the protein in cancer are yet to be established. Here, we show that elafin elicits pro-apoptotic effects in melanoma cells but not in normal melanocytes. Elafin triggered the intrinsic apoptotic pathway as evidenced by the increased caspase 9 activity and unaltered caspase 8 activity. Caspase 9-specific siRNA, but not caspase 8-specific siRNA, dramatically abrogated elafin-induced apoptosis. Elevated level of p53 was observed, resulting in increased transcriptional activation and consequent expression of downstream effector molecules (Bax, Puma, Noxa, p21). Moreover, the apoptotic effect of elafin was inhibited by p53-specific siRNA and the p53 inhibitor pifithrin-alpha. Elafin treatment of xenograft mice of melanoma cells led to significantly smaller tumor sizes compared with those of untreated control mice. Immunohistochemical analysis revealed decreased elafin expression in melanoma tissue specimens. Western blot and reverse transcription analyses indicated transcriptional repression of the elafin gene in melanoma cells. Our results collectively indicate that elafin induces apoptosis in melanoma cells through a p53-dependent intrinsic apoptotic pathway, and that repression of elafin expression in melanoma may contribute to disease progression.

Colonoscopic lesions in patients with spinal cord injury.

BACKGROUND/OBJECTIVE: In spinal cord injury (SCI), loss of central or peripheral neural control causes neurogenic bowel. Patients may not exhibit the typical signs and symptoms of gastrointestinal disease. Few studies have looked at the risk of gastrointestinal disease in this group and the indications for preventive screening. The objective of this study was to study colonoscopic lesions in patients with SCI and determine whether there are any differences in the prevalence of lesions between SCI and control patients. DESIGN: Case control study. METHODS: Twenty-five patients with SCI were compared with 41 control patients who received colonoscopy at the same time. Mann-Whitney test for continuous variable, and Fisher exact test for frequency variables were used. OUTCOME MEASURES: Demographic information, duration of SCI, and colonoscopy findings were gathered. RESULTS: Colonic lesions were observed in 52% of patients with SCI and in 41.5% of control patients. Most frequent lesions in SCI group were inflammatory bowel disease (16%) and polyp (16%), followed by proctitis (12%) and hemorrhoid (12%). In the control group, hemorrhoid (17.1%) was most common, followed by polyp (12.2%) and melanosis coli (9.8%). No significant differences were found between the 2 groups. In the SCI group, no significant differences in lesions were found among the patients with cervical, thoracic, and lumbar SCI in the SCI group. Duration of SCI did not affect the pattern of colonoscopic lesions. CONCLUSION: Patients with SCI had the same incidence of colonscopic lesions as control patients. Inflammatory bowel disease, which is a risk factor for cancer, was the most common findings in the SCI group, although there was no significant difference from the control group. In patients with SCI, colonoscopy screening is warranted at the same frequency as for the general population.

Circadian rhythm of surfactant protein A, B and C mRNA in rats.

BACKGROUND: All organisms have developed an internal timing system capable of reacting to and anticipating environmental stimuli with a program of appropriately timed metabolic, physiologic and behavioral events. The alveolar epithelial type II cell of the mammalian lung synthesizes, stores, and secretes a lipoprotein pulmonary surfactant, which functions to stabilize alveoli at low lung volumes. METHODS: The authors investigated the diurnal variation of surfactant protein A, B and C mRNA accumulation. The diunal variation on gene expression of surfactant protein A, B and C was analysed using filter hybridization at 9 a.m., 4 p.m. and 11 p.m. Lung SP-A protein content was determined by double sandwich ELISA assay using a polyclonal antiserum raised in rabbits against purified rat SP-A. RESULTS: 1. The accumulation of SP-A mRNA at 4 p.m. was significantly decreased by 23.5% compared to the value at 9 a.m. (p < 0.05). 2. The accumulation of SP-B mRNA at 4 p.m. and 11 p.m. was decreased by 15.1% and 5.7%, respectively, compared to the value at 9 a.m. (p = 0.07, p = 0.69). 3. The accumulation of SP-C mRNA at 4 p.m. and 11 p.m. was decreased by 6.8% and 7.7%, respectively, compared to the value at 9 a.m. (p = 0.38, p = 0.57). 4. Total lung SP-A content at 4 p.m. and 11 p.m. was increased by 5.3% and 15.9%, respectively, compared to the value at 9 a.m. (p = 0.64, p = 0.47). CONCLUSION: These findings represent the diurnal variation of surfactant proteins mRNA expression in vivo. These results indicated that the diurnal variation of significant gene expression is observed in hydrophilic surfactant protein rather than in hydrophobic surfactant proteins.