Functional characterization of O-methyltransferases used to catalyse site-specific methylation in the post-tailoring steps of pradimicin biosynthesis.

AIMS: To identify the roles of the two O-methyltransferase homologous genes pdmF and pdmT in the pradimicin biosynthetic gene cluster of Actinomadura hibisca P157-2. METHODS AND RESULTS: Pradimicins are pentangular polyphenol antibiotics synthesized by bacterial type II polyketide synthases (PKSs) and tailoring enzymes. Pradimicins are naturally derivatized by combinatorial O-methylation at two positions (i.e., 7-OH and 11-OH) of the benzo[α]naphthacenequinone structure. PdmF and PdmT null mutants (PFKO and PTKO) were generated. PFKO produced the 11-O-demethyl shunt metabolites 11-O-demethylpradimicinone II (1), 11-O-demethyl-7-methoxypradimicinone II (2), 11-O-demethylpradimicinone I (3) and 11-O-demethylpradimicin A (4), while PTKO generated the 7-O-demethyl derivatives pradimicinone II (5) and 7-hydroxypradimicin A (6). Pradimicinones 1, 2, 3, and 5 were fed to a heterologous host Escherichia coli harbouring expression plasmid pET-22b::pdmF or pET-28a::pdmT. PdmF catalysed 11-O-methylation of pradimicinones 1, 2, and 3 regardless of O-methylation at the C-7 position, while PdmT was unable to catalyse 7-O-methylation when the C-11 hydroxyl group was methylated (5). CONCLUSIONS: PdmF and PdmT were involved in 11-O- and 7-O-methylations of the benzo[α]naphthacenequinone moiety of pradimicin, respectively. Methylation of the C-7 hydroxyl group precedes methylation of the C-11 hydroxyl group in pradimicin biosynthesis. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first reported demonstration of the functions of PdmF and PdmT for regiospecific O-methylation, which contributes to better understanding of the post-PKS modifications in pradimicin biosynthesis as well as to rational engineering of the pradimicin biosynthetic machinery.

[The timing of renal replacement therapy on mortality in patients with acute renal injury: a reanalysis of two recently published randomized control trials].

To investigate the sources of inconsistent findings between two randomized control trials["initiation strategies for renal-replacement therapy in the intensive care unit"(AKIKI trial) vs"effect of early vs delayed initiation of renal replacement therapy on mortality in critically ill patients with acute kidney injury"(ELAIN trial) ], regarding"timing of renal replacement therapy (RRT) on mortality in patients with acute kidney injury (AKI). By reanalysis of the published data, it was found that demographics, severity of primary disease and stage of AKI before initiation of RRT were quite different between AKIKI and ELAIN trials. Interestingly, similar mortalities were demonstrated in late group of ELAIN trial, both of early and late groups of AKIKI trial [all patients were classified at Kidney Disease: Improving Global Outcomes (KDIGO) classification stage 3 of AKI, P>0.05] although a significant reduction of mortality was determined in early group of ELAIN trial (KDIGO stage 2 of AKI).Therefore, it was concluded that inconsistent results were largely attributable to the heterogeneity of enrolled patients between ELAIN vs AKIKI trials, including demographics and severity of AKI(AKI stage) before initiation of RRT.

Application of emitter-sample hybrid terahertz time-domain spectroscopy to investigate temperature-dependent optical constants of doped InAs.

We investigate temperature-dependent carrier dynamics of InAs crystal by using reflection-type terahertz time-domain spectroscopy, particularly with a recently developed emitter-sample hybrid structure. We successfully obtain the optical conductivity in a terahertz frequency of bulk InAs whose dc conductivity is in the range of 100-150 Ω-1 cm-1. We find that both real and imaginary parts of the optical conductivity can be fit well with the simple Drude model, and the free-carrier density and the scattering rate obtained from the fit are in good agreement with corresponding values obtained by using other techniques, such as the Hall measurement and the dc-resistivity measurement. These results clearly demonstrate that the proposed technique of adopting the emitter-sample hybrid structure can be exploited to determine temperature-dependent optical constants in a reflection geometry and hence to investigate electrodynamics of bulk metallic systems.

Comparison of reversal with neostigmine of low-dose rocuronium vs. reversal with sugammadex of high-dose rocuronium for a short procedure.

Some short procedures require deep neuromuscular blockade, which needs to be reversed at the end of the procedure. Forty-four patients undergoing elective laryngeal micro-surgery were randomly allocated into two groups: rocuronium 0.45 mg.kg-1 with neostigmine (50 µg.kg-1 with glycopyrrolate 10 µg.kg-1 ) reversal (moderate block group) vs. rocuronium 0.90 mg.kg-1 with sugammadex (4 mg.kg-1 ) reversal (deep block group). The primary outcome was the intubating conditions during laryngoscopy secondary outcomes included recovery of neuromuscular block; conditions for tracheal intubation; satisfaction score as determined by the surgeon; onset of neuromuscular block; and postoperative sore throat. The onset of neuromuscular block was more rapid, and intubation conditions and ease of intra-operative laryngoscopy were more favourable, and the satisfaction score was lower in the moderate block group compared with the deep block group. No difference was found in the incidence of postoperative sore throat. In laryngeal micro-surgery, the use of rocuronium 0.9 mg.kg-1 with sugammadex for reversal was associated with better surgical conditions and a shorter recovery time than rocuronium 0.45 mg.kg-1 with neostigmine.

Specific PCR detection of Fusarium oxysporum f. sp. raphani: a causal agent of Fusarium wilt on radish plants.

Fusarium oxysporum, a causal agent of Fusarium wilt, is one of the most important fungal pathogens worldwide, and detection of F. oxysporum DNA at the forma specialis level is crucial for disease diagnosis and control. In this study, two novel F. oxysporum f. sp. raphani (For)-specific primer sets were designed, FOR1-F/FOR1-R and FOR2-F/FOR2-R, to target FOQG_17868 and FOQG_17869 ORFs, respectively, which were selected based on the genome comparison of other formae speciales of F. oxysporum including conglutinans, cubense, lycopersici, melonis, and pisi. The primer sets FOR1-F/FOR1-R and FOR2-F/FOR2-R that amplified a 610- and 425-bp DNA fragment, respectively, were specific to For isolates which was confirmed using a total of 40 F. oxysporum isolates. From infected plants, the FOR2-F/FOR2-R primer set directly detected the DNA fragment of For isolates even when the radish plants were collected in their early stage of disease development. Although the loci targeted by the For-specific primer sets were not likely involved in the pathogenesis, the primer set FOR2-F/FOR2-R is available for the determination of pathogenicity of radish-infecting F. oxysporum isolates. This study is the first report providing novel primer sets to detect F. oxysporum f. sp. raphani. SIGNIFICANCE AND IMPACT OF THE STUDY: Because plant pathogenic Fusarium oxysporum has been classified into special forms based on its host specificity, identification of F. oxysporum usually requires a pathogenicity assay as well as knowledge of the morphological characteristics. For rapid and reliable diagnosis, this study provides PCR primer sets that specifically detect Fusarium oxysporum f. sp. raphani (For) which is a devastating pathogen of radish plants. Because one of the primer sets directly detected the DNA fragment of For isolates from infected plants, the specific PCR method demonstrated in this study will provide a foundation for integrated disease management practices in commodity crops.

Antifungal activity of rimocidin and a new rimocidin derivative BU16 produced by Streptomyces mauvecolor BU16 and their effects on pepper anthracnose.

AIMS: The objective of this study was to explore antifungal metabolites targeting fungal cell envelope and to evaluate the control efficacy against anthracnose development in pepper plants. METHODS AND RESULTS: A natural product library comprising 3000 microbial culture extracts was screened via an adenylate kinase (AK)-based cell lysis assay to detect antifungal metabolites targeting the cell envelope of plant-pathogenic fungi. The culture extract of Streptomyces mauvecolor strain BU16 displayed potent AK-releasing activity. Rimocidin and a new rimocidin derivative, BU16, were identified from the extract as active constituents. BU16 is a tetraene macrolide containing a six-membered hemiketal ring with an ethyl group side chain instead of the propyl group in rimocidin. Rimocidin and BU16 showed broad-spectrum antifungal activity against various plant-pathogenic fungi and demonstrated potent control efficacy against anthracnose development in pepper plants. CONCLUSIONS: Antifungal metabolites produced by S. mauvecolor strain BU16 were identified to be rimocidin and BU16. The compounds displayed potent control efficacy against pepper anthracnose. SIGNIFICANCE AND IMPACT OF THE STUDY: Rimocidin and BU16 would be active ingredients of disease control agents disrupting cell envelope of plant-pathogenic fungi. The structure and antifungal activity of rimocidin derivative BU16 is first described in this study.

Single nucleotide polymorphisms of TNFAIP3 are associated with systemic lupus erythematosus in Han Chinese population.

The polymorphisms of tumour necrosis factor alpha-induced protein 3 (TNFAIP3) have been found to associate with several autoimmune diseases. This study aimed to explore the association of single nucleotide polymorphisms (SNPs) of TNFAIP3 gene with systemic lupus erythematosus (SLE) in Han Chinese. Thirty-two SNPs were genotyped in 284 patients with SLE and 630 controls using the ligation detection reaction (LDR) method. The quality control steps and statistical analyses were performed using the PLINK 1.07 package and HAPLOVIEW software. We found that 13 SNPs in TNFAIP3 showed significant association with SLE (P < 1.85 × 10(-3)), and all of them were in high linkage disequilibrium (LD). After conditioning on the SNP rs2230926, other 12 SNPs did not show association (P > 0.27). All 13 SNPs showed most significant association in the dominant model. In haplotype analysis, a long risk SNP haplotype (GCCCGTGTCATGG) showed most significant association (P = 1.00 × 10(-4)). In conclusion, our data suggest that TNFAIP3 is a susceptible gene for SLE in the Han Chinese population.

Two familial cases of Olmsted-like syndrome with a G573V mutation of the TRPV3 gene.

Olmsted syndrome (OS) is a rare disease, characterized by symmetrical, sharply defined, hyperkeratotic, mutilating plaques on the palms and soles, which are associated with periorificial keratotic plaques. Other clinical manifestations of OS include diffuse alopecia, leucokeratosis of the oral mucosa, onychodystrophy, hyperkeratotic linear streaks, follicular hyperkeratosis and constriction of the digits. A recent study identified de novo mutations in the gene for transient receptor potential vanilloid 3 (TRPV3), causing constitutive activation of the TRPV3 channel, as a cause of OS. We report familial inheritance of OS in a family from Mongolia, which was caused by a previously undescribed G573V point mutation in TRPV3. To date, mutations in the G573 residue of TRPV3 have been reported in seven cases of OS: G573S in five cases, and G573C and G573A mutations in one case each. We present a Mongolian familial case of G573V point mutation in TRPV3.

Self-expandable metal stents for malignant esophageal obstruction: a comparative study between extrinsic and intrinsic compression.

Self-expandable metal stents (SEMSs) are effective for malignant esophageal obstruction, but usefulness of SEMSs in extrinsic lesions is yet to be elucidated. This study is aimed at evaluating the clinical usefulness of SEMSs in the extrinsic compression compared with intrinsic. A retrospective review was conducted for 105 patients (intrinsic, 85; extrinsic, 20) with malignant esophageal obstruction who underwent endoscopic SEMSs placement. Technical and clinical success rates were evaluated and clinical outcomes were compared between extrinsic and intrinsic group. Extrinsic group was mostly pulmonary origin. Overall technical and clinical success rate was 100% and 91%, respectively, without immediate complications. Extrinsic and intrinsic group did not differ significantly in clinical success rate. The median stent patency time was 131.3 ± 85.8 days in intrinsic group while that of extrinsic was 54.6 ± 45.1 due to shorter survival after stent insertion. The 4-, 8-, and 12-week patency rates were 90.5%, 78.8%, and 64.9% respectively in intrinsic group, while stents of extrinsic group remained patent until death. Uncovered, fully covered, and double-layered stent were used evenly and the types did not influence patency in both groups. In conclusion, esophageal SEMSs can safely and effectively be used for malignant extrinsic compression as well as intrinsic.

Serum 25-hydroxyvitamin D level and the risk of mild cognitive impairment and dementia: the Korean Longitudinal Study on Health and Aging (KLoSHA).

OBJECTIVE: The association of low vitamin D status with mild cognitive impairment (MCI), a preclinical condition that can lead to dementia, has not yet been fully explored. Our aim was to investigate the association between vitamin D status and the future risk of MCI and dementia in older adults. DESIGN, SETTING AND PARTICIPANTS: We conducted a population-based prospective study as a part of the Korean Longitudinal Study on Health and Aging. Four hundred and twelve elderly participants who completed evaluations of cognitive function and metabolic parameters in 2005-2006 and 2010-2011 were analysed. MAJOR OUTCOME MEASURE: The rate of development of MCI or dementia during the study period was compared according to baseline vitamin D status. Binary logistic regression analysis was performed to investigate any independent association between vitamin D status and the risks of MCI or dementia. RESULTS: Among 405 subjects that remained after excluding seven demented subjects at baseline, 338 subjects remained unchanged or improved in their diagnosis for cognitive function during the study period, whereas 67 subjects showed progression to MCI or dementia. When analyzing 236 subjects whose baseline mini-mental state examination (MMSE) scores were <27, severe vitamin D deficiency at baseline, defined as <25 nmol/l, was independently associated with the progression of cognitive impairment. Among 297 subjects who were normal at baseline, 50 acquired MCI and 247 remained normal. Severe vitamin D deficiency was also independently associated with the development of MCI when analyzing 145 subjects whose baseline MMSE scores were <27. CONCLUSION: Severe vitamin D deficiency was independently associated with the future risk of MCI as well as dementia, especially in older adults whose baseline MMSE scores had decreased only modestly.

Extraction of optical constants using multiple reflections in the terahertz emitter-sample hybrid structure.

To extract optical constants of nontransmitted samples in the terahertz (THz) spectral region, we employ a THz emitter-sample hybrid structure where THz pulses are generated at the emitter surface and multiply reflected at the interface between the THz emitter and the sample. Since each THz electric field profile appears well separated in a time domain, we could obtain the amplitude and phase spectra for each pulse from the Fourier transform, and determine the optical constants of the sample numerically based on the Fresnel equations. We applied this technique for doped semiconductors, and found that obtained optical constants are in good agreement with the values determined by using other conventional THz techniques.

Computational fluid dynamics simulation of airflow in the trachea and main bronchi for the subjects with left pulmonary artery sling.

BACKGROUND: Left pulmonary artery sling (LPAS) is a rare but severe congenital anomaly, in which the stenoses are formed in the trachea and/or main bronchi. Multi-detector computed tomography (MDCT) provides useful anatomical images, but does not offer functional information. The objective of the present study is to quantitatively analyze the airflow in the trachea and main bronchi of LPAS subjects through computational fluid dynamics (CFD) simulation. METHODS: Five subjects (four LPAS patients, one normal control) aging 6-19 months are analyzed. The geometric model of the trachea and the two main bronchi is extracted from the MDCT images. The inlet velocity is determined based on the body weight and the inlet area. Both the geometric model and personalized inflow conditions are imported into CFD software, ANSYS. The pressure drop, mass flow ratio through two bronchi, wall pressure, flow velocity and wall shear stress (WSS) are obtained, and compared to the normal control. RESULTS: Due to the tracheal and/or bronchial stenosis, the pressure drop for the LPAS patients ranges 78.9-914.5 Pa, much higher than for the normal control (0.7 Pa). The mass flow ratio through the two bronchi does not correlate with the sectional area ratio if the anomalous left pulmonary artery compresses the trachea or bronchi. It is suggested that the C-shaped trachea plays an important role on facilitating the air flow into the left bronchus with the inertia force. For LPAS subjects, the distributions of velocities, wall pressure and WSS are less regular than for the normal control. At the stenotic site, high velocity, low wall pressure and high WSS are observed. CONCLUSIONS: Using geometric models extracted from CT images and the patient-specified inlet boundary conditions, CFD simulation can provide vital quantitative flow information for LPAS. Due to the stenosis, high pressure drops, inconsistent distributions of velocities, wall pressure and WSS are observed. The C-shaped trachea may facilitate a larger flow of air into the left bronchus under the inertial force, and decrease the ventilation of the right lung. Quantitative and personalized information may help understand the mechanism of LPAS and the correlations between stenosis and dyspnea, and facilitate the structural and functional assessment of LPAS.

Automatic pulmonary fissure detection and lobe segmentation in CT chest images.

BACKGROUND: Multi-detector Computed Tomography has become an invaluable tool for the diagnosis of chronic respiratory diseases. Based on CT images, the automatic algorithm to detect the fissures and divide the lung into five lobes will help regionally quantify, amongst others, the lung density, texture, airway and, blood vessel structures, ventilation and perfusion. METHODS: Sagittal adaptive fissure scanning based on the sparseness of the vessels and bronchi is employed to localize the potential fissure region. Following a Hessian matrix based line enhancement filter in the coronal slice, the shortest path is determined by means of Uniform Cost Search. Implicit surface fitting based on Radial Basis Functions is used to extract the fissure surface for lobe segmentation. By three implicit fissure surface functions, the lung is divided into five lobes. The proposed algorithm is tested by 14 datasets. The accuracy is evaluated by the mean (±S.D.), root mean square, and the maximum of the shortest Euclidian distance from the manually-defined fissure surface to that extracted by the algorithm. RESULTS: Averaged over all datasets, the mean (±S.D.), root mean square, and the maximum of the shortest Euclidian distance are 2.05 ± 1.80, 2.46 and 7.34 mm for the right oblique fissure. The measures are 2.77 ± 2.12, 3.13 and 7.75 mm for the right horizontal fissure, 2.31 ± 1.76, 3.25 and 6.83 mm for the left oblique fissure. The fissure detection works for the data with a small lung nodule nearby the fissure and a small lung subpleural nodule. The volume and emphysema index of each lobe can be calculated. The algorithm is very fast, e.g., to finish the fissure detection and fissure extension for the dataset with 320 slices only takes around 50 seconds. CONCLUSIONS: The sagittal adaptive fissure scanning can localize the potential fissure regions quickly. After the potential region is enhanced by a Hessian based line enhancement filter, Uniform Cost Search can extract the fissures successfully in 2D. Surface fitting is able to obtain three implicit surface functions for each dataset. The current algorithm shows good accuracy, robustness and speed, may help locate the lesions into each lobe and analyze them regionally.

Development and characterization of new microsatellites for walnut (Juglans regia).

The expressed sequence tag (EST) database represents a potentially valuable resource for the development of simple sequence repeat (SSR) markers for use in evolutionary studies. EST-SSRs reveal polymorphisms not only within the source taxon, but in related taxa as well. In this paper, we describe a case study in which the publicly available walnut (Juglans regia) EST database was used to develop SSR markers for use in the genetic analysis of the widespread Juglans nigra and Carya cathayensis and an endangered species Annamocarya sinensis. A total of 7262 unigenes, including 1911 contigs and 5351 singletons, were obtained from 13,559 ESTs retrieved from the NCBI database. The 7262 unigenes were further reduced to 706 EST-SSR sequences containing 805 SSR loci. Then, 309 EST-SSR primers were randomly designed, and 77 were identified with five high across-species transferability cross-species: namely, J. regia, J. nigra, C. cathayensis, Carya dabieshanensis, and A. sinensis. Thirteen highly polymorphic EST-SSRs were further used for genetic analyses in these above five species.

Conservation genetics of Annamocarya sinensis (Dode) Leroy, an endangered endemic species.

The endangered perennial plant Annamocarya sinensis (Dode) Leroy is a tertiary relict tree restricted to southeastern China and northern Vietnam. To explore endangerment mechanisms, develop protection strategies, and guide reintroduction efforts for this species, we investigated genetic diversity and population structure by surveying 70 individuals from three distinct populations using 12 polymorphic microsatellite markers. We found high genetic diversity for A. sinensis as indicated by high allelic diversity (allelic number = 4.667 ± 0.436, effective number of alleles = 2.913 ± 0.249), excess heterozygosity (observed heterozygosity = 0.586 ± 0.039, expected heterozygosity = 0.582 ± 0.029), and low fixation index (-0.028 ± 0.057). Our research revealed low genetic differentiation (FST = 0.066 ± 0.011) and no correlation between genetic distance and geographic distance. Analysis of molecular variance attributed 87% of the variance to differences within the population, whereas 13% was distributed among populations. The protection strategy should aim to protect as many populations as possible. Promoting sexual reproduction among various genotypes and establishing an outcrossing program are advisable for A. sinensis.

Study on transient photocurrent induced by energy level defect of schottky diode irradiated by high power pulsed laser.

The transient photocurrent is one of the key parameters of the spatial radiation effect of photoelectric devices, and the energy level defect affects the transient photocurrent. In this paper, by studying the deep level transient spectrum of a self-designed Schottky diode, the defect properties of the interface region of the anode metal AlCu and Si caused by high-temperature annealing at 150 â„ƒ, 200 â„ƒ and 300 â„ƒ for 1200 h have been quantitatively analyzed. The study shows that the defect is located at the position of + 0.41 eV on the valence band, the concentration is 2.8 [Formula: see text] 1013/cm2, and the capture cross section is [Formula: see text] = 8.5 [Formula: see text] 1017. The impurity energy level mainly comes from the diffusion of Al atom in anode metal. We found that the defect did not cause the electrical performance degradation and obvious morphology change of the device, but the transient photocurrent increased significantly. The reason is that the high temperature treatment results in a growth in the density of states at the interface between AlCu-Si. The more mismatched dislocations and recombination center increased the reverse current of the heterojunction. The above view is proved by the TCAD simulation test.

[Cystic echinococcosis of the waist and hip: a case report].

A patient with cystic echinococcosis was presented with primary lesions in the waist and hip. The case was misdiagnosed as subcutaneous abscess at initial diagnosis, and then definitively diagnosed as echinococcosis by means of imaging examinations and anti-Echinococcus antibody test. This case was reported with aims to improve the awareness of cystic echinococcosis among clinical physicians to avoid and reduce the misdiagnosis and missing diagnosis.

Metabolic syndrome-related SNPs in HLA and TNF7L2 may be risk factors for generalized pustular psoriasis in Chinese Han population.

Background: Generalized pustular psoriasis (GPP) is a rare and severe type of psoriasis. Previous studies have reported that metabolic syndrome and its components have been associated with psoriasis. Objective: To investigate the association of metabolic syndrome-related single-nucleotide polymorphisms (SNPs) and GPP in Chinese Han population. Materials and Methods: One hundred and thirty-six (136) GPP patients and 965 healthy controls were recruited in the study. Approximately, 4 ml peripheral venous blood was collected from each participant. After collection, second-generation sequencing was used to detect genetic polymorphism of 15 SNPs. The plink 1.07 software package was used for statistical analysis. Results: Rs805303 (p = 0.01, OR = 0.70) and rs3177928 (p = 3.18E-07, OR = 2.66) in HLA were significantly different between the two groups. Moreover, rs4506565 (p = 1.41E-03, OR = 2.72) and rs7901695 (p = 9.39E-04, OR = 2.82) in TCF7L2 were significantly associated with GPP in patients without a previous history of PsV. Genotype analysis of rs4506565 and rs7901695 showed that under the recessive model, genotype frequencies of rs4506565 (p = 0.00, OR = 18.52) and rs7901695 (p = 0.00, OR = 18.44) were significantly different between GPP patients and healthy controls. Conclusion: Rs805303 and rs3177928 in HLA may increase the risk of GPP in the Chinese Han population. TCF7L2 may be a risk factor for GPP in patients without a previous history of PsV.