Construction of Escherichia coli K-12 Strain Deficient in relA and spoT Using the λ Red Site-Specific Recombinase System

Previous studies have shown that an exposure to sub-inhibitory levels of kanamycin induces capsule synthesis which confers antibiotic resistance. To examine the role of spoT and relA genes in capsule synthesis and acquired physiological antibiotic resistance during sub-inhibitory kanamycin treatment, multiple studies were performed but showed inconclusive results. A potential explanation for the lack of success in the past was the use of genetic mutants containing a number of uncharacterized mutations, which resulted in limited comparability. In this study, the λ Red recombinase system has been utilized to construct an E. coli K-12 ΔrelAΔspoT mutant by disrupting the spoT gene in E. coli K-12 ΔrelA strain, JW2755-3. A PCR fragment was generated using primers with homologous sequences found upstream and downstream of the spoT gene, and a template plasmid, pKD3, carrying chloramphenicol resistance gene that is flanked by FLP recognition target sites. This product was electroporated into the strain JW2755-3 expressing the λ Red recombinase system. Five chloramphenicolresistant transformants were selected and PCR-tested for the presence of new junctions and locus-specific fragments. Construct SL11W447-4 had the correct structure as evidenced by four PCR tests performed using various combinations of locus-and cat-specific primers. This work provides a basis for future studies

Hyperspectral Analysis of Pine Wilt Disease to Determine an Optimal Detection Index

Bursaphelenchus xylophilus, the pine wood nematode (PWN) which causes pine wilt disease, is currently a serious problem in East Asia, including in Japan, Korea, and China. This paper investigates the hyperspectral analysis of pine wilt disease to determine the optimal detection indices for measuring changes in the spectral reflectance characteristics and leaf reflectance in the Pinus thunbergii (black pine) forest on Geoje Island, South Korea. In the present study, we collected the leaf reflectance spectra of pine trees infected with pine wilt disease using a hyperspectrometer. We used 10 existing vegetation indices (based on hyperspectral data) and introduced the green-red spectral area index (GRSAI). We made comparisons between non-infected and infected trees over time. A t-test was then performed to find the most appropriate index for detecting pine wilt disease-infected pine trees. Our main result is that, in most of the infected trees, the reflectance changed in the red and mid-infrared wavelengths within two months after pine wilt infection. The vegetation atmospherically resistant index (VARI), vegetation index green (VIgreen), normalized wilt index (NWI), and GRSAI indices detected pine wilt disease infection faster than the other indices used. Importantly, the GRSAI results showed less variability than the results of the other indices. This optimal index for detecting pine wilt disease is generated by combining red and green wavelength bands. These results are expected to be useful in the early detection of pine wilt disease-infected trees

Photometric Properties of the Near-contact Binary GW Geminorum

New multiband CCD photometry is presented for the eclipsing binary GW Gem; the $RI$ light curves are the first ever compiled. Four new minimum timings have been determined. Our analysis of eclipse timings observed during the past 79 years indicates a continuous period increase at a fractional rate of +(1.2$\pm$0.1)$\times10^{-10}$, in excellent agreement with the value $+1.1\times10^{-10}$ calculated from the Wilson-Devinney binary code. The new light curves display an inverse O'Connell effect increasing toward longer wavelengths. Hot and cool spot models are developed to describe these variations but we prefer a cool spot on the secondary star. Our light-curve synthesis reveals that GW Gem is in a semi-detached, but near-contact, configuration. It appears to consist of a near-main-sequence primary star with a spectral type of about A7 and an evolved early K-type secondary star that completely fills its inner Roche lobe. Mass transfer from the secondary to the primary component is responsible for the observed secular period change.Comment: 20 pages, including 5 figures and 6 tables, accepted for publication in PAS

Application of the Savitzky-Golay Filter to Land Cover Classification Using Temporal MODIS Vegetation Indices

In this study, the Savitzky-Golay filter was applied to smooth observed unnatural variations in the temporal profiles of the Normalized Difference Vegetation Index (NDVI} and the Enhanced Vegetation Index {EVI} time series from the MODerate Resolution Imaging Spectroradiometer (MODIS}. We computed two sets of land cover classifications based 011 the NDVI and EVI time series before and after applying the Savitzky-Golay filter. The resulting classification from the filtered versions of the vegetation indices showed a substantial improvement in accuracy when compared to the classifications from the unfiltered versions. The classification by the EVIsg had the highest K (0.72} for all classes compared to those of the EVI (0.67}, NDVI (0.63}, and NDV/sg (0.62). Therefore, we conclude that the EVIsg is best suited for land cover classification compared to the other data sets in this study

Calcium Uptake and Release through Sarcoplasmic Reticulum in the Inferior Oblique Muscles of Patients with Inferior Oblique Overaction

We characterized and compared the characteristics of Ca2+ movements through the sarcoplasmic reticulum of inferior oblique muscles in the various conditions including primary inferior oblique overaction (IOOA), secondary IOOA, and controls, so as to further understand the pathogenesis of primary IOOA. Of 15 specimens obtained through inferior oblique myectomy, six were from primary IOOA, 6 from secondary IOOA, and the remaining 3 were controls from enucleated eyes. Ryanodine binding assays were performed, and Ca2+ uptake rates, calsequestrins and SERCA levels were determined. Ryanodine bindings and sarcoplasmic reticulum Ca2+ uptake rates were significantly decreased in primary IOOA (p<0.05). Western blot analysis conducted to quantify calsequestrins and SERCA, found no significant difference between primary IOOA, secondary IOOA, and the controls. Increased intracellular Ca2+ concentration due to reduced sarcoplasmic reticulum Ca2+ uptake may play a role in primary IOOA

Phosphorylation of Histone H2A.X in Peripheral Blood Mononuclear Cells May Be a Useful Marker for Monitoring Cardiometabolic Risk in Nondiabetic Individuals

Phosphorylation of H2A.X (serine 139) in the histone H2A family located in the downstream of the DNA damage kinase signaling cascade is an important indicator of DNA damage. Recently, phosphorylation of H2A.X was proposed as a sensitive biomarker of aging. This study investigated if phosphorylation of H2A.X in peripheral blood mononuclear cells (PBMCs) is associated with cardiometabolic risk in nondiabetic individuals. Basic parameters and oxidative stress/inflammatory markers were measured in nondiabetic healthy Koreans (n = 119). Phosphorylation of H2A.X was measured randomly among the study subjects using a flow cytometer. According to the number of metabolic syndrome risk factor (MetS-RF), the study subjects were subdivided into &quot;super healthy&quot; (MetS − RF = 0, n = 71) and &quot;MetS-risk&quot; (MetS − RF ≥ 1, n = 48) groups. Phosphorylation of H2A.X in PBMCs (percentages and mean fluorescence intensity) was significantly higher in the MetS-risk group than in the super healthy group after adjusting for age, sex, cigarette smoking, and alcohol consumption. Phosphorylated H2A.X was positively correlated with the number of MetS-RF as well as waist circumference, blood pressures, triglyceride, Hb A1C , oxidized LDL, high sensitivity C-reactive protein, tumor necrosis factor-alpha, and alanine aminotransferase after the adjustment. The present study suggested that phosphorylated H2A.X in circulating PBMCs measured by flow cytometer may be a useful marker for monitoring cardiometabolic risk in nondiabetic individuals

Clinical and molecular epidemiology of human bocavirus in respiratory and fecal samples from children in Hong Kong

Background. Human bocavirus (HBoV) is a recently discovered parvovirus associated with respiratory tract infections in children. We conducted the first systematic prospective clinical and molecular study using nasopharyngeal aspirates (NPAs) and fecal samples. Methods. NPAs negative for influenza virus, parainfluenza virus, respiratory syncytial virus, adenovirus, and coronavirus and fecal samples from patients with acute gastroenteritis were included. On the basis of results from a pilot study using 400 NPAs from all age groups, a prospective 12-month study was conducted to detect HBoV in 1200 NPAs and 1435 fecal samples from patients <18 years old by polymerase chain reaction. The complete genome sequences of HBoVs from 12 NPAs and 12 fecal samples were determined. Results. Of the 400 NPAs collected in the pilot study, 20 (5.0%) were found to contain HBoV, all from children <5 years old. In the subsequent prospective study of pediatric patients, HBoV was detected in 83 (6.9%) of 1200 NPAs. Upper and lower respiratory tract infections were equally common. HBoV was detected in 30 (2.1%) of 1435 fecal samples. Fever and watery diarrhea were the most common symptoms. The seasonality of HBoV in NPAs and fecal samples was similar. Codetection with other pathogens occurred in 33% and 56% of NPAs and fecal samples, respectively, from patients with HBoV infection. Genomes of HBoVs from NPAs and fecal samples displayed minimal sequence variations. Conclusions. HBoV was detected in fecal specimens in children with acute gastroenteritis. A single lineage of HBoV was associated with both respiratory tract and enteric infections. © 2007 by the Infectious Diseases Society of America. All rights reserved.published_or_final_versio

E-Cadherin/β-Catenin Complex and the Epithelial Barrier

E-Cadherin/β-catenin complex plays an important role in maintaining epithelial integrity and disrupting this complex affect not only the adhesive repertoire of a cell, but also the Wnt-signaling pathway. Aberrant expression of the complex is associated with a wide variety of human malignancies and disorders of fibrosis resulting from epithelial-mesenchymal transition. These associations provide insights into the complexity that is likely responsible for the fibrosis/tumor suppressive action of E-cadherin/β-catenin

Successful Treatment of Tracheal Stenosis with Slide Tracheoplasty after the Failure of Resection with End-to-End Anastomosis

The combined effects of inhaled irritant gases and heat in burn patients can result in the development of laryngotracheal strictures. Several factors could adversely affect the development of tracheal stenosis and cause the growth of granulation tissue. Yet the current treatment options for this condition are limited because of the paucity of case reports. We report here on a case of a patient who experienced recurrent upper tracheal stenosis after an inhalation injury. She displayed repetitive symptoms of stenosis even after several laryngomicrosurgeries and resection with end-to-end anastomosis. Finally, 5 yr after the burn injury, slide tracheoplasty was successfully performed and the postoperative check-up findings and the increased airway volume seen on imaging were all satisfactory

Effect of ankle taping on the ankle muscle strength in young healthy women

Of the various approaches to manage or prevent injuries on the ankle joint, Kinesio taping improves static posture stability, stimulating the neuromuscular control system. This study aims to investigate of ankle taping on the peak torque and average power of ankle muscle in young heathy women. Taping was applied to the extensor digitorum, tibialis anterior, gastrocnemius, soleus of the ankle in 16 healthy women. Isokinetic measurement of the dorsiflexion and plantarflexion patterns were recorded before and after taping. As a result of this study, the pre-post isokinetic parameters improved significantly for plantarflexion (p<0.05) while those for dorsiflexion did not. Through this study, it was found out that ankle taping improved the muscle power of the plantarflexor of young healthy women, and it is expected that applying ankle taping would help young healthy women