Programmable site-selective labeling of oligonucleotides based on carbene catalysis

Site-selective modification of oligonucleotides serves as an indispensable tool in many fields of research including research of fundamental biological processes, biotechnology, and nanotechnology. Here we report chemo- and regioselective modification of oligonucleotides based on rhodium(I)-carbene catalysis in a programmable fashion. Extensive screening identifies a rhodium(I)-catalyst that displays robust chemoselectivity toward base-unpaired guanosines in single and double-strand oligonucleotides with structurally complex secondary structures. Moreover, high regioselectivity among multiple guanosines in a substrate is achieved by introducing guanosine-bulge loops in a duplex. This approach allows the introduction of multiple unique functional handles in an iterative fashion, the utility of which is exemplified in DNA-protein cross-linking in cell lysates. The methods for investigation of DNA-binding proteins require site-selective chemical modifications to be introduced into oligonucleotides. Here, the authors report a chemo- and regioselective method for the modification of unpaired guanosines in single- and double-stranded oligonucleotides, based on Rh(I)-carbene catalysis

Trends in Cancer Screening Rates among Korean Men and Women: Results from the Korean National Cancer Screening Survey (KNCSS), 2004-2010

PURPOSE: The Korean National Cancer Screening Survey (KNCSS) is a continuous nationwide survey implemented by the National Cancer Center in Korea since 2004. The purpose of the present study was to report trends in cancer screening rates for the five major cancers (stomach, liver, colorectal, breast, and cervix uteri) in Korean men and women. MATERIALS AND METHODS: The study used KNCSS data collected between 2004 and 2010. The survey was conducted on Korean men aged 40-74 years and Korean women aged 30-74 years with no history of cancer diagnosis. The annual percentage change and corresponding 95% confidence intervals were used to examine changes in annual screening rates. RESULTS: Screening rates with recommendation increased by 4.4% annually for stomach cancer, 1.5% for liver cancer, 2.8% per year for colorectal cancer, 4.5% for breast cancer, and 1.2% for cervix uteri cancer. The increasing trend in cancer screening rates, with the exception of liver cancer, was significant. CONCLUSION: Cancer screening rates have increased consistently from 2004 to 2010 among Korean men and women. Stomach and breast cancer screening rates in particular have increased markedly.ope

Evaluation of therapeutic effects of natural killer (NK) cell-based immunotherapy in mice using in vivo apoptosis bioimaging with a caspase-3 sensor

Natural killer (NK) cellâ based immunotherapy is a promising strategy for cancer treatment, and caspaseâ 3 is an important effector molecule in NK cellâ mediated apoptosis in cancers. Here, we evaluated the antitumor effects of NK cellâ based immunotherapy by serial noninvasive imaging of apoptosis using a caspaseâ 3 sensor in mice with human glioma xenografts. Human glioma cells expressing both a caspaseâ 3 sensor as a surrogate marker for caspaseâ 3 activation and Renilla luciferase (Rluc) as a surrogate marker for cell viability were established and referred to as D54â CR cells. Human NK92 cells were used as effector cells. Treatment with NK92 cells resulted in a timeâ and effector numberâ dependent increase in bioluminescence imaging (BLI) activity of the caspaseâ 3 sensor in D54â CR cells in vitro. Caspaseâ 3 activation by NK92 treatment was blocked by Zâ VAD treatment in D54â CR cells. Transfusion of NK92 cells induced an increase of the BLI signal by caspaseâ 3 activation in a doseâ and timeâ dependent manner in D54â CR tumorâ bearing mice but not in PBSâ treated mice. Accordingly, sequential BLI with the Rluc reporter gene revealed marked retardation of tumor growth in the NK92â treatment group but not in the PBSâ treatment group. These data suggest that noninvasive imaging of apoptosis with a caspaseâ 3 sensor can be used as an effective tool for evaluation of therapeutic efficacy as well as for optimization of NK cellâ based immunotherapy.â Lee, H. W., Singh, T. D., Lee, S.â W., Ha, J.â H., Rehemtulla, A., Ahn, B.â C., Jeon, Y.â H., Lee, J. Evaluation of therapeutic effects of natural killer (NK) cellâ based immunotherapy in mice using in vivo apoptosis bioimaging with a caspaseâ 3 sensor. FASEB J. 28, 2932â 2941 (2014). www.fasebj.orgPeer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/154541/1/fsb2fj13243014.pd

Molecular cloning of the Ecotin gene in Escherichia coli

AbstractThe nucleotide sequence of a 876 bp region in E. coli chromosome that encodes Ecotin was determined. The proposed coding sequence for Ecotin is 486 nucleotides long, which would encode a protein consisting of 162 amino acids with a calculated molecular weight of 18 192 Da. The deduced primary sequence of Ecotin includes a 20-residue signal sequence, cleavage of which would give rise to a mature protein with a molecular weight of 16 099 Da. Ecotin does not contain any consensus reactive site sequences of known serine protease inhibitor families, suggesting that Ecotin is a novel inhibitor

Cytoprotective Activity of Glycyrrhizae radix Extract Against Arsenite-induced Cytotoxicity

Licorice, Glycyrrhizae radix, is one of the herbal medicines in East Asia that has been commonly used for treating various diseases, including stomach disorders. This study investigated the effect of licorice on arsenite (As)-induced cytotoxicity in H4IIE cells, a rat hepatocyte-derived cell line. Cell viability was significantly diminished in As-treated H4IIE cells in a time and concentration-dependent manner. Furthermore, results from flow cytometric assay and DNA laddering in H4IIE cells showed that As treatment induced apoptotic cell death by activating caspase-3. Licorice (0.1 and 1.0 mg ml−1) treatment significantly inhibited cell death and the activity of caspase-3 in response to As exposure. These results demonstrate that licorice induced a cytoprotective effect against As-induced cell death by inhibition of caspase-3

Raman Scattered HeII 6545 in the Young and Compact Planetary Nebula NGC 6790

We present the high resolution spectra of the youn and compact planerary nebula NGC 6790 obtained with the echelle spectrograph at Bohyunsan Optical Astronomy Observatory and report the discovery of Raman scattered He II 6545 in this object. This line feature is formed in a thick neutral region surrounding the hot central star, where He II 1025 line photons are scattered inelastically by hydrogen atoms. A Monte Carlo technique is adopted to compute the line profile with a simple geometric model, in which the neutral region is in the form of a cylindrical shell that is expanding from the central star. From our line profile analysis, the expansion velocity of the HI region lies in the range V_exp = 15- 19 Km/s. Less stringent constraints are put on the HI column density N_HI and covering factor C, where the total flux of Raman He II 6545 is consistent with the product of N_HI and C being CN_HI ~ 0.5 X 10^20 cm^-2. The Monte Carlo profiles from stationary emission models exhibit deficit in the wing parts. A much better fit is obtained when the He II emission region is assumed to take the form of a ring that slowly rotates with a rotation speed ~ 18Km/s. Brief discussions are presented regarding the mass loss processes and future observations

Nest entry shape change may cause nest abandonment in urban cavity-nesting species: a case study of the Tree Sparrow Passer montanus

The threat of predation is the main cause of bird nest abandonment, with such behaviour imposing considerable energetic costs on breeding birds. However, for several species, nest abandonment can be a less costly alternative to complete brood failure. In this study, we examined nest abandonment among Eurasian Tree Sparrows (Passer montanus) by surveying 71 Tree Sparrow nests with various types of entry holes and conducted artificially manipulating some of the entrance shapes. We found that nest abandonment was caused by changes to the nest entry shape in seven cases and by human interference in two cases. Nest abandonments occurred throughout the breeding season, and breeding pairs attempted to breed again immediately after nest abandonment. The results of the artificial nest entry shape manipulation experiment showed that nine of twelve nests (75.0%) were abandoned where the nest entrance holes were widened, and six of eleven nests (54.5%) were abandoned where the nest entrance holes were narrowed. However, none of the nests were abandoned where the entry shape was unchanged. Thus, nest abandonment by Tree Sparrows is correlated with nest entry shape manipulation and is more likely to occur when the energy cost of breeding again is less than that of abandoning the nest