Therapeutic Effects of Inhibitor of ompA Expression against Carbapenem-Resistant Acinetobacter baumannii Strains.

The widespread of carbapenem-resistant Acinetobacter baumannii (CRAB) is of great concern in clinical settings worldwide. It is urgent to develop new therapeutic agents against this pathogen. This study aimed to evaluate the therapeutic potentials of compound 62520, which has been previously identified as an inhibitor of the ompA promoter activity of A. baumannii, against CRAB isolates, both in vitro and in vivo. Compound 62520 was found to inhibit the ompA expression and biofilm formation in A. baumannii ATCC 17978 at sub-inhibitory concentrations in a dose-dependent manner. These inhibitory properties were also observed in clinical CRAB isolates belonging to sequence type (ST) 191. Additionally, compound 62520 exhibited a bacteriostatic activity against clinical clonal complex (CC) 208 CRAB isolates, including ST191, and ESKAPE pathogens. This bacteriostatic activity was not different between STs of CRAB isolates. Bacterial clearance was observed in mice infected with bioimaging A. baumannii strain 24 h after treatment with compound 62520. Compound 62520 was shown to significantly increase the survival rates of both immunocompetent and neutropenic mice infected with A. baumannii ATCC 17978. This compound also increased the survival rates of mice infected with clinical CRAB isolate. These results suggest that compound 62520 is a promising scaffold to develop a novel therapeutic agent against CRAB infections.

Neuroprotective properties of ethanolic extract of Citrus unshiu Markovich peel through NADPH oxidase 2 inhibition in chemotherapy-induced neuropathic pain animal model.

The present study aimed to determine the antioxidant effect of Citrus unshiu Markovich (CUM) extract in neuronal cell lines under oxidative stress and to investigate the effect of chemotherapy-induced peripheral neuropathy (CIPN) on the nociceptive response in a preclinical mice model. We tested the inhibition of H2 O2 in Neuro2A cells treated with CUM. Experimental animals were treated with oxaliplatin to induce CINP, and then administered oral CUM for 4 weeks in order to observe the effect of CUM. Animals were evaluated weekly for thermal hyperalgesia and digital motor nerve conduction velocity (NCV). Lumbar dorsal root ganglia (DRG) isolated from each animal were evaluated through immunochemical and western blot analysis for nerve damage, inflammatory response, and expression of redox signaling factors. The main mechanisms were determined to be decreased inducible nitric oxide synthase (iNOS) production due to the inhibition of NADPH oxidase 2 (NOX2). To determine the functional role of NOX2 in CINP, we administrated CUM into NOX2-deficient mice with neuropathic pain. Therefore, we suggest that CUM controls the expression levels of inflammatory factors in CINP via NOX2 inactivation. This study demonstrated that a complementary medicine such as CUM might be a potential novel therapeutic agent for the treatment of CINP.

PGC-1α, a potential therapeutic target against kidney aging.

Aging is defined as changes in an organism over time. The proportion of the aged population is markedly increasing worldwide. The kidney, as an essential organ with a high energy requirement, is one of the most susceptible organs to aging. It is involved in glucose metabolism via gluconeogenesis, glucose filtration and reabsorption, and glucose utilization. Proximal tubular epithelial cells (PTECs) depend on lipid metabolism to meet the high demand for ATP. Recent studies have shown that aging-related kidney dysfunction is highly associated with metabolic changes in the kidney. Peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α), a transcriptional coactivator, plays a major role in the regulation of mitochondrial biogenesis, peroxisomal biogenesis, and glucose and lipid metabolism. PGC-1α is abundant in tissues, including kidney PTECs, which demand high energy. Many in vitro and in vivo studies have demonstrated that the activation of PGC-1α by genetic or pharmacological intervention prevents telomere shortening and aging-related changes in the skeletal muscle, heart, and brain. The activation of PGC-1α can also prevent kidney dysfunction in various kidney diseases. Therefore, a better understanding of the effect of PGC-1α activation in various organs on aging and kidney diseases may unveil a potential therapeutic strategy against kidney aging.

Comparison of DNA extraction methods for drug susceptibility testing by allele-specific primer extension on a microsphere-based platform: Chelex-100 (in-house and commercialized) and MagPurix TB DNA Extraction Kit.

Tuberculosis (TB), caused by infections of the Mycobacterium tuberculosis (MTB) complex, is the ninth leading cause of death worldwide, and several molecular approaches for MTB species identification and the detection of mutations associated with drug resistance have been developed to date. We previously developed a diagnostic assay for drug susceptibility testing that can detect mutations conferring resistance to anti-TB drugs using allele-specific primer extension on a microsphere-based platform for multiplex polymerase chain reaction. The aim of the present study was to optimize this diagnostic assay based on the evaluation of three methods for extracting mycobacterial DNA from clinical samples. Mycobacterial DNA of 81 samples was digested and decontaminated by N-acetyl-l-cysteine-2% NaOH and then extracted using three methods: "in-house" 5% Chelex-100 chelating resin, InstaGene Matrix, and MagPurix TB DNA Extraction Kit. The former two methods are manual extraction methods, whereas the MagPurix TB DNA Extraction Kit is an automated extraction method used with the MagPurix 12 s automated nucleic acid purification system. The extracted DNA was then subjected to our diagnostic assay, and the results were compared among methods. The magnetic bead method exhibited a higher extraction efficiency and resulted in greater diagnostic efficacy than the two resin-based methods with respect to both target gene detection and acid-fast bacilli smear grades. Therefore, the MagPurix TB DNA Extraction Kit is the optimal MTB DNA extraction method for our diagnostic assay of TB drug susceptibility testing.

Aerosol-assisted plasma deposition of hydrophobic polycations makes surfaces highly antimicrobial.

The currently used multistep chemical synthesis for making surfaces antimicrobial by attaching to them hydrophobic polycations is replaced herein by an aerosol-assisted plasma deposition procedure. To this end, N,N-hexyl,methyl-PEI (HMPEI) is directly plasma-coated onto a glass surface. The resultant immobilized HMPEI coating has been thoroughly characterized and shown to be robust, bactericidal against Escherichia coli, and virucidal against human influenza virus.

Pedobacter namyangjuensis sp. nov. isolated from soil and reclassification of Nubsella zeaxanthinifaciens Asker et al. 2008 as Pedobacter zeaxanthinifaciens comb. nov.

A Gram-stain-negative, non-motile, strictly aerobic, yellow-pigmented bacterium, designated strain 5G38(T), was isolated from a field cultivated with Chinese cabbage in Korea. The strain grew at 5-40°C and at pH 6.0-8.0. 16S rRNA gene sequence analysis revealed that strain 5G38(T) represented a distinct lineage within the family Sphingobacteriaceae and showed the highest 16S rRNA gene sequence similarity of 95.2% with Pedobacter koreensis WPCB189(T), followed by Pedobacter agri PB92(T) (94.6%), Pedobacter suwonensis 15-52(T) (94.4%), Pedobacter rhizosphaerae 01-96(T) (94.4%), Pedobacter sandarakinus DS-27(T) (94.4%), and Nubsella zeaxanthinifaciens TDMA-5(T) (94.3%). Strain 5G38(T) formed monophyletic clade with Nubsella zeaxanthinifaciens in the cluster comprised of species of the genus Pedobacter. Chemotaxonomic characteristics of the novel strains, including DNA G+C content of genomic DNA (37.0 mol%), the predominant respiratory quinine (MK-7), and the major fatty acids which were iso-C15:0, summed feature 3 (comprising C16:1 ω7c and/or iso-C15:0 2-OH) and iso-C17:0 3-OH, are similar to those of the genus Pedobacter. However, the novel strains can be distinguished from the other species of Pedobacter by physiological properties. The name Pedobacter namyangjuensis sp. nov. is therefore proposed for strain 5G38(T) (KACC 13938(T) =NBRC 107692(T)) as the type strain. Furthermore, the reclassification of Nubsella zeaxanthinifaciens as Pedobacter zeaxanthinifaciens comb. nov. is proposed.

Surgical treatment of dermatomal capillary malformations in the adult face.

BACKGROUND: Facial capillary malformations (CMs) rarely recede; they often become darker and raised in proportion to their growth. These malformations may hypertrophy in adulthood, resulting in increased disfigurement and dysfunction. Laser treatment is considered a first-line therapy for focal CMs, but thick wide lesions, which are accompanied by hypertrophy and have a well-circumscribed nodularity, may be treated with surgical excision and reconstruction. METHODS: We retrospectively reviewed the records of 25 consecutive patients who had undergone complete or partial excisions of facial capillary malformations in our unit. After the excisions, the defects that encompassed their facial aesthetic units were subsequently covered by various methods, including primary closures, local flaps, expanded flaps, split-thickness skin grafts, and full thickness skin grafts. RESULTS: The data demonstrated satisfactory results and reliability. Our patients were treated without significant complications, and all of the patients were moderately or fully satisfied with the outcome of their surgeries. CONCLUSIONS: Among the many reconstructive options for adult patients with facial capillary malformations, thick split-thickness skin grafts can be a good choice for the coverage of widely excised wounds.

Combined AlloDerm® and thin skin grafting for the treatment of postburn dyspigmented scar contracture of the upper extremity.

Postburn dyspigmented scar contractures of the upper extremity often require aesthetic improvement. The ideal reconstruction of this deformity remains a challenge because the various available skin grafts and flaps result in skin colour mismatches, prominent marginal scars and donor morbidity. Postburn scar contractures and dyspigmented areas of the upper extremity can be improved by a combination of dermabrasion and Alloderm(®) graft over scar-releasing defect. Their raw surfaces are subsequently re-surfaced with thin split-thickness skin graft (0.005-0007 inches thick). Twenty-seven patients with wide dyspigmented scar contractures of the upper extremity underwent the combined techniques described by us. The median patient age at burn incidents was 3 years and at operation was 24 years. Median thin skin graft area was 180cm(2), and the median AlloDerm(®) graft area was 40cm(2). Thin skin and AlloDerm(®) grafts took root completely in all patients without re-grafting. Follow-up periods ranged from 30 to 67 months (average 47.6 months). Re-pigmentation was achieved in all cases and all scar contractures were adequately released and treated with an AlloDerm(®) graft. Paired differences between preoperative and postoperative parameters as determined by the Vancouver Scar Scale (VSS) were significant. Focal hypertrophic scar and reddish-coloured graft sites gradually improved over 3-4 years postoperatively. Graft margin and donor scars were inconspicuous. Our described combined technique was found to treat these deformities effectively. We suggest that the use of Alloderm(®) and thin skin grafting be considered in patients concerned about this type of cosmetic disfigurement.

Rudaea cellulosilytica gen. nov., sp. nov., isolated from soil.

A yellow-pigmented, Gram-negative, aerobic, rod-shaped bacterium, strain KIS3-4T, was isolated from soil collected on Daechung Island in the West Sea of Korea. Phylogenetic analysis based on the 16S rRNA gene sequence placed strain KIS3-4T in a distinct lineage in the family Xanthomonadaceae. Strain KIS3-4T shared 87.3-93.7% sequence similarity with members of the family Xanthomonadaceae, and was related most closely to the genera Dyella and Dokdonella. In its biochemical characteristics, strain KIS3-4T was clearly separable from other genera within the family Xanthomonadaceae on the basis of the hydrolysis of cellulose and urea, high G+C content (64 mol%) and fatty acid profile. Major fatty acids (>10% of the total fatty acids) were iso-C17:1omega9c (32.8%), iso-C17:0 (18.0%) and iso-C16:0 (12.7%). Q-8 was the predominant respiratory quinone. Phosphatidylethanolamine and several unidentified aminophospholipids and phospholipids were present. Based on its unique phenotypic, genotypic and phylogenetic features, strain KIS3-4T represents a novel genus and species, for which the name Rudaea cellulosilytica gen. nov., sp. nov. is proposed. The type strain of Rudaea cellulosilytica is KIS3-4T (=KACC 12734T=JCM 15422T).

Quantitative improvement of 16S rDNA DGGE analysis for soil bacterial community using real-time PCR.

Denaturing gradient gel electrophoresis (DGGE) of 16S rDNA fragments has been frequently used to profile a structure of the bacterial community in a given soil. However, this procedure has various types of intrinsic error and bias, thus often misleads the relative abundance of bacterial populations. In order to establish a reliability for the current 16S rDNA DGGE method, we investigated various parameters and potential sources of errors in the DGGE procedures, such as primer mismatch, dNTP concentration, DNA polymerase, PCR cycles, uneven amplification of templates, secondary structure of PCR product, melting domain profiles, and acrylamide/bis concentration. Our result showed that the relative band intensities of the corresponding 16S rDNA templates were closely correlated with the differences of the melting temperature between the higher and lower melting domains of the PCR products. In addition, application of i) real-time PCR, ii) combination of PCR primers and iii) optimization of both dNTP and acrylamide/bis concentrations significantly improved the quantitative representation of bacterial 16S rDNA levels in the mixed samples. Especially, identification of the inflection points of DNA samples through the real-time PCR was crucial for the accurate representation of soil bacterial populations. Beyond these points DNA templates can be over-amplified to a saturated level independently of their initial amounts. Therefore for the accurate analysis of soil bacterial community, a quantitative 16S rDNA DGGE analysis needs to be performed in combination with a real-time PCR.

Chitinophaga niabensis sp. nov. and Chitinophaga niastensis sp. nov., isolated from soil.

Two yellow-coloured bacterial strains, designated JS13-10(T) and JS16-4(T), were isolated from soil from Jeju Island, Republic of Korea. On the basis of 16S rRNA gene sequence analysis, the strains were found to be affiliated with members of the genus Chitinophaga. Phenotypically, the novel strains were identified as being different from each other and from recognized species of the genus Chitinophaga. DNA-DNA hybridization tests between the two novel strains and closely related Chitinophaga reference strains produced DNA relatedness values that were significantly lower (<36 %) than those generally accepted as the highest threshold for the phylogenetic definition of a species. On the basis of their distinct taxonomic characteristics, these strains represent two novel species of the genus Chitinophaga, for which the names Chitinophaga niabensis sp. nov. (type strain JS13-10(T)=KACC 12952(T)=JCM 15440(T)) and Chitinophaga niastensis sp. nov. (type strain JS16-4(T)=KACC 12954(T)=JCM 15441(T)) are proposed.

Niabella ginsengisoli sp. nov., isolated from soil cultivated with Korean ginseng.

The taxonomic status of a yellow- to light orange-coloured strain isolated from soil of a Korean ginseng field was established based on a polyphasic investigation. The novel isolate, strain GR10-1(T), was an obligately aerobic, Gram-staining-negative, non-motile, flexirubin-pigment-producing, short rod-shaped bacterium. The strain grew optimally at 28-30 degrees C, at pH 7.0 and in the presence of 0-1 % NaCl. Phylogenetic analyses based on 16S rRNA gene sequences demonstrated that the new isolate showed the highest sequence similarities with Niabella aurantiaca R2A15-11(T) (95.1 %) and Niabella soli JS13-8(T) (94.6 %). The DNA G+C content of strain GR10-1(T) was 43 mol%. It contained iso-C(15 : 1) G (36.4 %) and iso-C(15 : 0) (32.8 %) as the major fatty acids (>10 %) and MK-7 as the major isoprenoid quinone. On the basis of evidence from our polyphasic taxonomic study, it was concluded that strain GR10-1(T) should be classified within a novel species of the genus Niabella, for which the name Niabella ginsengisoli sp. nov. is proposed. The type strain is GR10-1(T) (=KACC 13021(T) =JCM 15444(T)).

Aquitalea denitrificans sp. nov., isolated from a Korean wetland.

A novel bacterium, designated strain 5YN1-3(T), was isolated from wetland peat collected from Yongneup, Korea. The bacterium was facultatively anaerobic, Gram-negative, yellow-coloured, rod-shaped, mesophilic and motile with one polar flagellum. The strain grew optimally at 30 degrees C, at pH 6.0-9.0 and with 0-1 % NaCl (w/v). 16S rRNA gene sequence analysis showed the highest similarity to the sequence from Aquitalea magnusonii TRO-001DR8(T), with 98.7 % sequence similarity. However, strain 5YN1-3(T) showed DNA-DNA relatedness of 43 % (40 % in a reciprocal experiment) with A. magnusonii LMG 23054(T). The strain contained summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c) and C(16 : 0) as major cellular fatty acids. On the basis of DNA-DNA relatedness and physiological and biochemical characterization, strain 5YN1-3(T) should be assigned to a novel species of the genus Aquitalea, for which the name Aquitalea denitrificans sp. nov. is proposed. The type strain is 5YN1-3(T) (=KACC 12729(T) =DSM 21300(T)).

Anti-obesity effect of Pinellia ternata extract in Zucker rats.

Pinellia ternata is known as the herb effective in removing dampness-phlegm, one of the causes of obesity in traditional Korean medicine. Pinellia ternata water extract (PE) was fed to rats after mixing with diet once a day (400 mg x kg(-1)) for 6 weeks. We investigated its effect on the thermogenesis and fatty acids oxidation with obese Zucker rats. We also determined the gene expression of uncoupling protein 1 (UCP1), peroxisome proliferators-activated receptor alpha (PPARalpha), and PPARgamma coactivator 1alpha (PGC1alpha). The PE treatment lowered the levels of triglyceride and free fatty acids (p<0.05) in blood of the obese rats and the body weight was also reduced slightly. It was also observed that PE significantly increased the expression of both UCP1 mRNA in brown adipose tissue (BAT) (p<0.001) and PPARalpha and PGC1alpha mRNA in white visceral adipose tissue (WAT) (p<0.05 and p<0.001, respectively), which may cause a reduction of obesity. These results suggested that PE would be able to affect anti-obesity through thermogenesis and fatty acid oxidation.

Homogeneous nucleation of n-propanol, n-butanol, and n-pentanol in a supersonic nozzle.

We have measured the nucleation conditions of n-propanol, n-butanol, and n-pentanol in a supersonic Laval nozzle, and estimated that the maximum nucleation rate J is 5 x 10(16) cm(-3) s(-1) with an uncertainty factor of 2. Plotting the vapor pressures p(J(max) ) and temperatures T(J(max) ) corresponding to the maximum nucleation rate as ln(p) versus 1T, produces a series of well separated straight lines. When these values are scaled by their respective critical parameters, p(c) and T(c), the data lie close to a single straight line. Comparing the experimental data to the predictions of classical nucleation theory reveals much higher experimental rates, and the deviation increases with increasing alcohol chain length and decreasing temperature. A scaling analysis in terms of Hale's scaled nucleation model [Phys. Rev. A 33, 4156 (1986); Metall. Trans. A 23, 1863 (1992)], clearly shows that our data are consistent with experimental nucleation rates measured using other devices that have characteristic rates many orders of magnitude lower.

Scale-up of Artemisia annua L. hairy root cultures produces complex patterns of terpenoid gene expression.

Hairy roots grow quickly, reach high densities, and can produce significant amounts of secondary metabolites, yet their scale-up to bioreactors remains challenging. Artemisia annua produces a rich array of terpenoids, including the sesquiterpene, artemisinin, and transformed roots of this species provide a good model for studying terpenoid production. These cultures were examined in shake flasks and compared with cultures grown in two types of bioreactors, a mist reactor and a bubble column reactor, which provide very different environments for the growing roots. Mist reactors have been shown previously to result in cultures that produce significantly more artemisinin per gram fresh weight of culture, while bubble column reactors have produced greater biomass. We have compared expression levels of four key terpenoid biosynthetic genes: 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), 1-deoxy-D-xylulose-5-phosphate synthase (DXS), 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), and farnesyl diphosphate synthase (FPS) in the three culture conditions. In shake flasks we found that although all four genes showed temporal regulation, only FPS expression correlated with artemisinin production. Light also affected the transcription of all four genes. Although expression in reactors was equivalent to or greater than that of roots grown in shake flasks, no correlation was found between expression level within six different zones of each reactor and their respective oxygen levels, light, and root-packing density. Surprisingly, transcriptional regulation of HMGR, DXS, DXR, and FPS was greatly affected by the position of the roots in each reactor. Thus, relying on a single reactor sample to characterize the gene activity in a whole reactor can be misleading, especially if the goal is to examine the difference between reactor types or operating parameters, steps essential in scaling up cultures for production.