Hrant M. Tovmassian
Full Text Available Los grupos compactos de galaxias Shakhbazian son las configuraciones más densas que se conocen. Hace algunos años iniciamos un estudio fotométrico y espectroscópico de estos grupos. En este artículo presentamos los resultados de la investigación en los grupos ShCG 104, ShCG 120, ShCG 243 y ShCG 245. Presentamos los corrimientos al rojo de las galaxias miembro, los resultados de la fotometría R, las curvas de brillo superficial-radio efectivo, las masas estimadas, las luminosidades y las razones masa-luminosidad de los grupos, además de algunos parámetros dinámicos como la dispersión de velocidad radial y el tiempo de cruce. Los ShCGs estudiados consisten principalmente de galaxias elípticas y lenticulares. Se muestra que algunas galaxias en estos grupos están en proceso de interacción.
The availability of synthetic oligodeoxynucleotides provides the opportunity to study the effects of 125I in DNA with added precision and detail, compared to an earlier study (Martin and Haseltine, Science 214, 296, 1981). The authors have designed a template/primer system which enables incorporation of 125IdC into a defined location in a 31mer. The main advantage of this approach is that relatively large amounts (a few μCi) of DNA with both 32P and 125I can be produced quite easily. Consequently, when 125I-induced DNA damage is analyzed on DNA sequencing gels, the distribution of 32P-labeled cleavage products can be determined more accurately, following accumulation of decay events under various conditions (e.g. temperature, buffer components, free radical scavengers). The results of preliminary experiments suggest that the yield of DNA strand breaks per decay approaches one, at least for the 125I labeled strand. Extension of this experimental approach will also enable investigation of the nature of the terminii (e.g. phophoryl-versus phophorylglycollate) at either side of 125I decay-induced DNA strand breaks. The question of energy migration is also being investigated in the oligodeoxynucleotide system, by studying the effects of inclusion of BrUdR nucleotides at various distances from the 125IdC
Full Text Available Abstract Background Persistent infection with hepatitis C virus (HCV is a leading cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. Standard therapy consists of a combination of interferon-alpha and ribavirin, but many patients respond poorly, especially those infected with HCV genotypes 1 and 4. Furthermore, standard therapy is associated with severe side-effects. Thus, alternative therapeutic approaches against HCV are needed. Findings Here, we studied the effect of a new class of antiviral agents against HCV, short, partially double-stranded oligodeoxynucleotides (ODNs, on viral replication. We targeted the 5’ nontranslated region (5’ NTR of the HCV genome that has previously been shown as effective target for small interfering RNAs (siRNAs in vitro. One of the investigated ODNs, ODN 320, significantly and efficiently reduced replication of HCV replicons in a sequence-, time- and dose-dependent manner. ODN 320 targets a genomic region highly conserved among different HCV genotypes and might thus be able to inhibit a broad range of genotypes and subtypes. Conclusions ODNs provide an additional approach for inhibition of HCV, might be superior to siRNAs in terms of stability and cellular delivery, and suitable against HCV resistant to standard therapy. This study underlines the potential of partially double-stranded ODNs as antiviral agents.
Cole Laurence A
Abstract Background hCG is a term referring to 4 independent molecules, each produced by separate cells and each having completely separate functions. These are hCG produced by villous syncytiotrophoblast cells, hyperglycosylated hCG produced by cytotrophoblast cells, free beta-subunit made by multiple primary non-trophoblastic malignancies, and pituitary hCG made by the gonadotrope cells of the anterior pituitary. Results and discussion hCG has numerous functions. hCG promotes progesterone p...
Rosson, Reinhardt A.; Maurina-Brunker, Julie; Langley, Kim; Pynnonen, Christine M.
A rapid, sensitive test using chemiluminescent oligodeoxynucleotide probes has been developed for detecting, identifying, and enumerating intact cells. The test is intended especially for use in detecting and enumerating bacteria and yeasts in potable water. As in related tests that have been developed recently for similar purposes, the oligodeoxynucleotide probes used in this test are typically targeted at either singlecopy deoxyribonucleic acid (DNA) genes (such as virulence genes) or the multiple copies (10,000 to 50,000 copies per cell) of 16S ribosomal ribonucleic acids (rRNAs). Some of those tests involve radioisotope or fluorescent labeling of the probes for reporting hybridization of probes to target nucleic acids. Others of those tests involve labeling with enzymes plus the use of chemiluminescent or chromogenic substrates to report hybridization via color or the emission of light, respectively. The present test is of the last-mentioned type. The chemiluminescence in the present test can be detected easily with relatively simple instrumentation. In developing the present test, the hybridization approach was chosen because hybridization techniques are very specific. Hybridization detects stable, inheritable genetic targets within microorganisms. These targets are not dependent on products of gene expression that can vary with growth conditions or physiological states of organisms in test samples. Therefore, unique probes can be designed to detect and identify specific genera or species of bacteria or yeast (in terms of rRNA target sequences) or can be designed to detect and identify virulence genes (genomic target sequences). Because of the inherent specificity of this system, there are few problems of cross-reactivity. Hybridization tests are rapid, but hybridization tests now available commercially lack sensitivity; typically, between 10(exp 6) and 10(exp 7) cells of the target organism are needed to ensure a reliable test. Consequently, the numbers of
Vlachova, J.; Ruzkova, M.; Jakoubkova, J.; Skrivan, J.; Zavadil, M.
The difference between the clinical finding and hCG levels in the serum is shown in two choriocarcinoma patients with widely progressing metastatic processes in the lungs. In the patient showing low hCG levels and poor chemotherapeutic response the presence is presumed of a free hCG-alpha subunit. The opinion is backed that negativity or relapse of a trophoblastic disease in the sensitive and specific hCG*-anti hCG-beta system should be watched because intact hCG is a dominant form of the hormone secerning due to the disease. It is admitted, however, that possible determination of the alpha and beta subunits of the hormone would greatly contribute to assessing the progress of the disease.
Bezvoda, Radek; Pleskot, Roman; Žárský, Viktor; Potocký, Martin
Vol. 1080. New York: Humana Press, 2014 - (Žárský, V.; Cvrčková, F.), s. 231-236. (Methods in Molecular Biology). ISBN 978-1-62703-643-6 R&D Projects: GA ČR GA13-19073S Institutional support: RVO:61389030 Keywords : Antisense oligodeoxynucleotide * Pollen tube * AODN Subject RIV: EB - Genetics ; Molecular Biology
2004 turned over maintainership of Prolog+ CG to the present author. In this paper, Prolog+CG is described in terms of its history, evolution, and maintenance. A special web-enabled version of Prolog+CG is also described. Possible interoperability with CGIF and the CharGer tool are explored. Finally......Prolog+CG is an implementation of Prolog with Conceptual Graphs as first-class datastructures, on a par with terms. As such, it lends itself well to applications in which reasoning with Conceptual Graphs and/or ontologies plays a role. It as originally developed by Prof. Dr. Adil Kabbaj, who in...
Tovmassian, H. M.; H. Tiersch; S. G. Navarro; V. H. Chavushyan; Tovmassian, G. H.; A. S. Amirkhanian; Neizvestny, S.
Los grupos compactos de galaxias Shakhbazian son las configuraciones más densas que se conocen. Hace unos pocos años iniciamos un estudio fotométrico y espectroscópico de estos grupos. En este artículo presentamos los resultados de la investigación en los grupos ShCG 31, ShCG 38, ShCG 43 y ShCG 282. Presentamos los corrimientos al rojo de las galaxias miembro, los resultados de la fonometría BV R, las curvas de brillo superficial-radio efectivo, las masas estimadas, luminosidad...
Tovmassian, Hrant M.; H. Tiersch; Tovmassian, Gaghik H.; V. H. Chavushyan; Neizvestny, S.; Pramskij, A. G.; J. P. Torres-Papaqui; M Rozas
Los grupos compactos de galaxias Shakhbazian son las configuraciones más densas que se conocen. Hace algunos años iniciamos un estudio fotométrico y espectroscópico de estos grupos. En este artículo presentamos los resultados de la investigación en los grupos ShCG 104, ShCG 120, ShCG 243 y ShCG 245. Presentamos los corrimientos al rojo de las galaxias miembro, los resultados de la fotometría R, las curvas de brillo superficial-radio efectivo, las masas estimadas, las luminosida...
Basermann, A.; Reichel, B.; Schelthoff, C. [Central Institute for Applied Mathematics, Juelich (Germany)
Conjugate gradient (CG) methods to solve sparse systems of linear equations play an important role in numerical methods for solving discretized partial differential equations. The large size and the condition of many technical or physical applications in this area result in the need for efficient parallelization and preconditioning techniques of the CG method. In particular for very ill-conditioned matrices, sophisticated preconditioner are necessary to obtain both acceptable convergence and accuracy of CG. Here, we investigate variants of polynomial and incomplete Cholesky preconditioners that markedly reduce the iterations of the simply diagonally scaled CG and are shown to be well suited for massively parallel machines.
Meng, Jie; Zhang, Linlin [Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, Shandong (China); Li, Li, E-mail: email@example.com [Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, Shandong (China); Li, Chunyan; Wang, Ting [Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, Shandong (China); University of Chinese Academy of Sciences, Beijing 100039 (China); Zhang, Guofan, E-mail: firstname.lastname@example.org [Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, Shandong (China)
Highlights: • CgMTF-1 and CgZnT1 were first identified in oysters. • CgMTF-1 localized in cell nucleus under unstressed conditions. • CgMTF-1 proteins could bind with the typical MRE motif. • CgMTF-1 activated CgZnT1, CgMT1 and CgMT4 promoters and regulated their expressions under zinc exposure. - Abstract: Oysters accumulate zinc at high tissue concentrations, and the metal response element (MRE)-binding transcription factor (MTF) functions as the cellular zinc sensor that coordinates the expression of genes involved in zinc efflux and storage, as well as those that protect against metal toxicity. In this study, we cloned MTF-1 in oysters and examined its regulation mechanism for its classic target genes, including MTs and ZnT1 under zinc exposure conditions. We cloned CgMTF-1 and determined the subcellular locations of its protein product in HEK293 cells. CgMTF-1 has a 2826 bp open reading frame that encodes a predicted polypeptide with 707 amino acid residues, showing six well-conserved zinc finger domains that are required for metal binding. In HEK293 cell lines, CgMTF-1 primarily localizes in the cell nucleus under unstressed conditions and nuclear translocation was not critical for the activation of this gene. We searched for CgMTF-1-regulated genes in oysters using RNA interference. Decreased expression levels of CgMT1, CgMT4, and CgZnT1 were observed after CgMTF-1 interference (>70% inhibition) under zinc exposure, indicating the critical role of CgMTF-1 in the regulation of these genes. We searched for a direct regulation mechanism involving CgMTF-1 for CgMT1, CgMT4, and CgZnT1 in vitro. EMSA experiments indicated that CgMTF-1 can bind with the MREs found in the CgZnT1, CgMT1 and CgMT4 promoter regions. Additionally, luciferase reporter gene experiments indicated that CgMTF-1 could activate the CgMT1, CgMT4, and CgZnT1 promoters. Overall, our results suggest that CgMTF-1 directly coordinates the regulation of CgMTs and CgZnT1 expression and plays
Objective To study the biological effects of cathepsin B phosporothioated antisense oligodeoxynucleotide on human osteosarcoma cell line MG-63 after transfection.Methods A 18-mer phosphorothioate antisense oligodeoxynucleotide(ASODN)targeted against the cathepsin B mRNA was transfected into the human osteosarcoma cell line MG-63 by lipofectamine 2000.The sense and nonsense oligodeoxynucleotides to cathepsin B and blank vector were used as controls.The expression of cathepsin B mRNA was examined by RT-PCR an...
S Agrawal; Mayrand, S H; Zamecnik, P. C.; Pederson, T
Oligodeoxynucleotides containing phosphodiester or modified internucleoside linkages were investigated with respect to their ability to be acted on by ribonuclease H activities present in a HeLa cell nuclear extract after hybridization with complementary sequences in RNA. Oligodeoxynucleotides complementary to nucleotides 2-14 of human U1 small nuclear RNA were investigated. Extensive cleavage of U1 RNA was observed with the unmodified oligodeoxynucleotide and with the phosphorothioate analog...
alternative glycosylation pattern on the free large hCG alpha in seminal plasma might reflect a modified function of this subunit in the male reproductive tract.
Cole Laurence A
Full Text Available Abstract Background hCG is a term referring to 4 independent molecules, each produced by separate cells and each having completely separate functions. These are hCG produced by villous syncytiotrophoblast cells, hyperglycosylated hCG produced by cytotrophoblast cells, free beta-subunit made by multiple primary non-trophoblastic malignancies, and pituitary hCG made by the gonadotrope cells of the anterior pituitary. Results and discussion hCG has numerous functions. hCG promotes progesterone production by corpus luteal cells; promotes angiogenesis in uterine vasculature; promoted the fusion of cytotrophoblast cell and differentiation to make syncytiotrophoblast cells; causes the blockage of any immune or macrophage action by mother on foreign invading placental cells; causes uterine growth parallel to fetal growth; suppresses any myometrial contractions during the course of pregnancy; causes growth and differentiation of the umbilical cord; signals the endometrium about forthcoming implantation; acts on receptor in mother's brain causing hyperemesis gravidarum, and seemingly promotes growth of fetal organs during pregnancy. Hyperglycosylated hCG functions to promote growth of cytotrophoblast cells and invasion by these cells, as occurs in implantation of pregnancy, and growth and invasion by choriocarcinoma cells. hCG free beta-subunit is produced by numerous non-trophoblastic malignancies of different primaries. The detection of free beta-subunit in these malignancies is generally considered a sign of poor prognosis. The free beta-subunit blocks apoptosis in cancer cells and promotes the growth and malignancy of the cancer. Pituitary hCG is a sulfated variant of hCG produced at low levels during the menstrual cycle. Pituitary hCG seems to mimic luteinizing hormone actions during the menstrual cycle.
H. M. Tovmassian
Full Text Available Los grupos compactos de galaxias Shakhbazian son las configuraciones más densas que se conocen. Hace unos pocos años iniciamos un estudio fotométrico y espectroscópico de estos grupos. En este artículo presentamos los resultados de la investigación en los grupos ShCG 31, ShCG 38, ShCG 43 y ShCG 282. Presentamos los corrimientos al rojo de las galaxias miembro, los resultados de la fonometría BV R, las curvas de brillo superficial-radio efectivo, las masas estimadas, luminosidades y las razones masa-luminosidad de los grupos, además de algunos parámetros dinámicos como la dispersión de velocidades radiales y el tiempo de cruce. Los ShCGs estudiados consisten principalmente de galaxias elípticas y lenticulares. Se muestra que algunas galaxias en estos grupos están en proceso de interacción.
Objective To evaluate the effect of human telomerase reverse transcriptase(hTERT) gene antisense oligodeoxynucleotide (ASON) on telomerase activity in K562 cells.Methods Telomerase activity was detemined by polymerase chain reaction enzyme-linked immunoassay (PCR-ELISA) in K562 cells treated with ASODN and hTERTmRNA expression was detected by reverse transcriptase polymerase chain reaction (RT-PCR). Results The hTERTmRNA level was decreased,and telomerase activity was significantly inhibited when the K562 cells were treated with ASODN for 48 h. Conclusion It is suggested that hTETR ASODN might specifically inhibit telomrase activity of K562 cells at translation level,and it is further proved that hTERT gene has significant correlation with telopmerase activity.
Huh, N; Rege, A A; Yoo, B; Kogan, T P; Kohn, H
Mitomycin C (1) is the prototypical bioreductive alkylating agent. Studies have shown that mitomycin C and its derivatives selectively alkylate guanine residues within di- and trinucleotide DNA sequences. This investigation sought to improve the selective DNA bonding properties of the mitomycins by coupling them with antisense oligodeoxynucleotides. Two procedures were developed that allowed the attachment of a phosphorothioate oligodeoxynucleotide containing a hexylamino spacer at the 5' terminus with a C(10)-activated mitomycin. In the first procedure, decarbamoylation of 1 (NaOCH3/ benzene) afforded 10-decarbamoylmitomycin C (10), which was treated with either dimethyl sulfate or methylthiochloroformate and base to yield 10-decarbamoylporfiromycin (11) and N(1a)-[(methylthio)-carbonyl]-10-decarbamoylmitomycin C (12), respectively. Activation of the C(10) site in 11 and 12 with 1,1'-carbonyldiimidazole or with 1,1'-thiocarbonyldiimidazole provided the N(1a)-substituted mitomycin 10-decarbamoyl-10-O-carbonylimidazoles (5, 7) and 10-decarbamoyl-10-O-thiocarbonylimidazoles (6, 8), respectively. Compounds 5-8 were reacted with glycine methyl ester hydrochloride (17) and base in both methylene chloride and aqueous buffered solutions to determine the ease and efficiency in which these C(10)-activated mitomycin derivatives coupled to amines. It was found that 5-8 all reacted with 17 in methylene chloride to give the coupled products 18-21 but that improved amine coupling yields in water were observed for the 10-decarbamoyl-10-O-thiocarbonylimidazoles 6 and 8 as compared with the 10-decarbamoyl-10-O-carbonylimidazoles 5 and 7. This finding led to the coupling of the phosphorothioate oligodeoxynucleotide, H2N(CH2)6-P(S)(OH)-GGCCCCGTG-GTGGCTCCAT (22) to 8. Compound 22 complemented a 19-base sequence in the translation initiation region of the human A-raf-1 gene. Use of excess 8 (28 equiv) with 22 gave only a 36% yield of the coupled product 23, which proved difficult to
The equations, coding, and procedures that are required to include a torus in the Combinatorial Geometry subroutines of the MORSE-CG code are described. The derivation and solutions of the quartic equation that describes a torus along with additional subroutines and the modifications to existing subroutines required to carry out the transport of neutrons and gamma rays in toroidal geometry are presented. The input requirements and a sample problem are included
Full Text Available Abstract The Herpes simplex virus (HSV is known as an infectious agent and widespread in the human population. The symptoms of HSV infections can range from mild to life threatening, especially in immune-compromised individuals. HSV infections are commonly treated with the guanosine analogue Aciclovir, but reports of resistance are increasing. Efforts are made to establish single-stranded antisense oligodeoxynucleotides (as and small interfering ribonucleic acids (siRNAs for antiviral treatment. Recently, another class of short interfering nucleic acids, partially double-stranded hairpin loop-structured 54 mer oligodeoxynucleotides (ODNs, was shown to allow hydrolysis of HIV RNA by binding to the viral RNA. This leads to a substrate for the viral RNase H. To assess the potential of such ODNs for inhibition of HSV-1 replication, five partially double-stranded ODNs were designed based on the sequences of known siRNAs against HSV-1 with antiviral activity. Three of them are directed against early and two against leaky late genes. Primary human lung fibroblasts, MRC-5, and African green monkey kidney cells, Vero, were transfected with ODNs and subsequently infected. The effect on HSV-1 replication was determined by analyzing the virus titer in cell culture supernatants by quantitative PCR and plaque assays. An inhibitory effect was observed with all five selected ODNs, with two cases showing statistical significance in both cell types. The observed effect was sequence-specific and dose dependent. In one case the ODN was more efficient than a previously described siRNA directed against the same target site in the mRNA of UL5, a component of the helicase/primase complex. HSV-1 virions and ODNs can be applied simultaneously without transfection reagent, but at a 50-fold higher concentration to Vero cells with similar efficiencies. The results underline the potential of partially double-stranded hairpin loop-structured ODNs as antiviral agents.
Ahuja, Kapil; Chang, Eun R; Gugercin, Serkan
Science and engineering problems frequently require solving a sequence of dual linear systems. Two examples are the Iterative Rational Krylov Algorithm (IRKA) for model reduction and Quantum Monte Carlo (QMC) methods in electronic structure calculations. This paper introduces Recycling BiCG, a BiCG method that recycles two Krylov subspaces from one pair of linear systems to the next pair. We develop an augmented bi-Lanczos algorithm and a modified two-term recurrence to include recycling in the iteration. The recycle spaces are approximate left and right invariant subspaces corresponding to the eigenvalues close to the origin. These recycle spaces are found by solving a small generalized eigenvalue problem alongside the dual linear systems being solved in the sequence. We test our algorithm in two application areas. First, we solve a discretized partial differential equation of convection-diffusion type, because these are well-known model problems. Second, we use Recycling BiCG for the linear systems arising ...
Glass-Mattie, Dana F [ORNL; McClanahan, Mark [Centers for Disease Control and Prevention; Koller, Loren [Environmental Health and Toxicology, Corvallis, Oregon; Adeshina, Femi [U.S. Environmental Protection Agency; Ross, Robert Hord [ORNL
The PAL protocol was applied to estimate inhalation exposure limits for phosgene (CG). Data on humans are limited to occupational exposures or accounts from the use of phosgene as a chemical warfare agent in WWI. Animal studies with phosgene show a steep dose-response curve for pulmonary edema and mortality with little species variability in effects. Although immediately upon exposure lacrimation and upper respiratory irritation can occur, the main effect in the target organ, a progressive pulmonary edema, occurs after a latency period of 1-24 hours. PAL estimates were approved by the Expert Consultation Panel for Provisional Advisory Levels in May 2007. Exposure limits for oral exposure to CG are not developed due to insufficient data. PAL estimates for inhalation exposure to CG are presented: The 24-hour PAL values for severity levels 1, 2, and 3 are 0.0017, 0.0033 and 0.022 ppm, respectively. The 30-day PAL values are 0.0006 and 0.0012 ppm for the PAL 1 and 2 values, respectively. These 30-day inhalation values were also accepted as the 90-day and 2-year PAL 1 and 2 values. Data were not available for deriving 30-day, 90-day and 2-year PAL 3 values.
Jendis, J; Strack, B; Volkmann, S; Böni, J; Mölling, K.
A previously described self-complementary oligodeoxynucleotide termed triplex-forming oligodeoxynucleotide (TFO A), 54 bases in length, designed against the polypurine tract of HIV-1 RNA, inhibited viral replication at a 1 to 3 microM concentration in acutely infected cells, whereas antisense and scrambled sequence oligodeoxynucleotides were ineffective. Three HIV-1 viral isolates from patients of clinical categories A1, B, and C3 were transmitted to peripheral blood mononuclear cells and tes...
Full Text Available CG8478 Cell line,Embryo,Pupae + dm3 http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/...target/CG8478.1.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/target/CG8478.5.tsv http://dbarchive.bios...ciencedbc.jp/kyushu-u/dm3/target/CG8478.10.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/colo/CG8478.Cell_line.tsv,http://dba...rchive.biosciencedbc.jp/kyushu-u/dm3/colo/CG8478.Embryo.tsv,http://dba...rchive.biosciencedbc.jp/kyushu-u/dm3/colo/CG8478.Pupae.tsv http://dbarchive.biosciencedbc.jp/kyus
A radioligand receptor assay system for urinary hCG and serum hCG was developed, using 1100--2000 xg fractions of homogenates of pig testis. This method is specific for hCG and LH, and the limit of detection for hCG was 20 mIU, which could be significantly discriminated from zero at the 95 percent confidence level. A nonspecific inhibition reaction which exerts its influence on the assay system has been found in the urine and serum, and in performing assays for urinary and serum hCG, it was necessary to remove the inhibition reaction by diluting the urine more than eight fold or subject the serum to an extraction procedure. Since the hCG of normal pregnancy and trophoblastic neoplasia showed a dose response curve similar to that of the International Standard hCG, it became clear that these hCG could be assayed with this method. The potency ratio RI measured by RRA and RIA of urinary hCG and serum hCG was different from those of normal pregnancy and trophoblastic neoplasia. (U.S.)
The VAM3D-CG computer code has been licensed for use at Hanford, from HydroGeologic, Inc., of Herndon, VA. Version 2.4b has been installed on the 3200GWW workstations, and is currently under configuration management. The purpose of this report is to describe the installation and configuration management of VAM3D-CG on the Hanford Computer System. VAM3D-CG is written in standard FORTRAN F77
The authors determined the levels of the CG contained in serum by RIA in 83 normal pregnant women and in 53 pregnant patients with ICP. The result showed that CG levels of ICP patients increased significantly as compared with that of normal middle and late pregnancy (P < 0.01). The routine examination of the serum CG levels in pregnant women is of great value of the earlier diagnosis to ICP and treatment as well as health care of perinatal period
Using anti-β-subunit hCG sera, radioimmunoassay (RIA) for hCG in urine and serum was developed, and its problems were examined. In the two measuring systems, the cross reaction between hCG and LH, and the non-specific inhibition of urine and serum were observed. For actual measurement, therefore, when a standard curve is prepared using urine or serum, and the measuring sensitivity is adjusted to 16 mIU/ml for urine hCG and to 8 mIU/ml for serum hCG, the effect of LH can be eliminated relatively, and the RIA obtained can be quite specific for hCG. When this method was applied clinically, the results were in agreement with the results from cases of trophoblastic tumors under treatment, those in remission, and menopausal females with high LH. Hardly any LH was detected in the cases in remission, menopausal females, or normal females. This method enables us to judge whether or not hCG-like substances are excreted from malignant tumors. In addition, it gives clearer information on the behavior of low level hCG in patients with trophoblastic tumors than does the pre-existing RIA for hCG. This method can be applied extensively to clinical practice. (Chiba, N.)
Costa, Catarina; Ribeiro, Jonathan; Miranda, Isabel M.; Silva-Dias, Ana; Cavalheiro, Mafalda; Costa-de-Oliveira, Sofia; Rodrigues, Acácio G.; Teixeira, Miguel C.
For years, antifungal drug resistance in Candida species has been associated to the expression of ATP-Binding Cassette (ABC) multidrug transporters. More recently, a few drug eﬄux pumps from the Drug:H+ Antiporter (DHA) family have also been shown to play a role in this process, although to date only the Candida albicans Mdr1 transporter has been demonstrated to be relevant in the clinical acquisition of antifungal drug resistance. This work provides evidence to suggest the involvement of the C. glabrata DHA transporters CgAqr1, CgQdr2, CgTpo1_1, and CgTpo3 in the clinical acquisition of clotrimazole drug resistance. A screening for azole drug resistance in 138 C. glabrata clinical isolates, from patients attending two major Hospitals in Portugal, was performed. Based on this screening, 10 clotrimazole susceptible and 10 clotrimazole resistant isolates were selected for further analysis. The transcript levels of CgAQR1, CgQDR2, CgTPO1_1, and CgTPO3 were found to be significantly up-regulated in resistant isolates when compared to the susceptible ones, with a level of correlation that was found to be similar to that of CgCDR2, an ABC gene known to be involved in the clinical acquisition of resistance. As a proof-of-concept experiment, the CgTPO3 gene was deleted in an azole resistant C. glabrata isolate, exhibiting high levels of expression of this gene. The deletion of CgTPO3 in this isolate was found to lead to decreased resistance to clotrimazole and fluconazole, and increased accumulation of azole drugs, thus suggesting the involvement of this transporter in the manifestation of azole resistance. PMID:27148215
He Maolin; Xiao Zengming; Li Shide; Chen Anmin
Objective To study the biological effects of cathepsin B phosporotbioated antisense oligodeoxynucleotide on human osteosarcoma cell line MG-63 after transfection. Methods A 18-mer phosphorothioate antisense oligodeoxynucleotide (ASODN) targeted against the cathepsin B mRNA was transfected into the human osteosarcoma cell line MG-63 by lipofectamine 2000. The sense and nonsense oligodeoxynucleotides to cathepsin B and blank vector were used as controls. The expression of cathepsin B mRNA was examined by RT-PCR and the expression of cathepsin B was examined by Western blot. The invasive capability of MG-63 cells was evaluated by the boydern chamber assay. Results The expression of cathcpsin B was obviously inhibited in antlsense oligodeoxynucleotide treated cells compared with the control cells. The number of invading MG-63 cells was significantly lower in the ASODN-treated groups than that in the control groups. Conclusion The cathepsin B ASODN significantly inhibits the expression of cathepsin B and invasive ability of MG-63 cell in osteosarcoma.
Full Text Available BACKGROUND: The development of mucosal vaccines is crucial to efficiently control infectious agents for which mucosae are the primary site of entry. Major drawbacks of these protective strategies are the lack of effective mucosal adjuvant. Synthetic oligodeoxynucleotides that contain several unmethylated cytosine-guanine dinucleotide (CpG-ODN motifs are now recognized as promising adjuvants displaying mucosal adjuvant activity through direct activation of TLR9-expressing cells. However, little is known about the efficacy of these molecules in stimulating the intestinal immune system in neonates. METHODOLOGY/PRINCIPAL FINDINGS: First, newborn mice received CpG-ODN orally, and the intestinal cytokine and chemokine response was measured. We observed that oral administration of CpG-ODN induces CXC and CC chemokine responses and a cellular infiltration in the intestine of neonates as detected by immunohistochemistry. We next compared the efficiency of the oral route to intraperitoneal administration in stimulating the intestinal immune responses of both adults and neonates. Neonates were more responsive to TLR9-stimulation than adults whatever the CpG-ODN administration route. Their intestinal epithelial cells (IECs indirectly responded to TLR9 stimulation and contributed to the CXC chemokine response, whereas other TLR9-bearing cells of the lamina-propria produced CC chemokines and Th1-type cytokines. Moreover, we showed that the intestine of adult exhibited a significantly higher level of IL10 at homeostasis than neonates, which might be responsible for the unresponsiveness to TLR9-stimulation, as confirmed by our findings in IL10-deficient mice. CONCLUSIONS/SIGNIFICANCE: This is the first report that deciphers the role played by CpG-ODN in the intestine of neonates. This work clearly demonstrates that an intraperitoneal administration of CpG-ODN is more efficient in neonates than in adults to stimulate an intestinal chemokine response due to their
Hickel, Gabriel R.; Vilas-Boas, José W. S.
In this work, we combine observations of optical linear polarization (R band), IRAS far-infrared images and radio molecular lines to investigate the cometary globule CG30 (at IRAS Vela Shell). CG30 shows Herbig-Haro objects, molecular outflows, hosts a very young binary star and has a star formation efficiency of about 6% to 17%. Its magnetic field is important to support the CG structure and shows evidence of torsion and compression of the field lines. The quadrupolar outflow of the binary star affects the temperature of the molecular gas, and changes the degree of polarization of the dust grains in the environment of CG30. This work is based on observations collected at LNA/CNPq, Brazil and SEST/ESO, Chile.
Chen, Rong; Luo, Xuefang; Di, Xin; Li, Ying; Sun, Yuqing; Hu, Yuzhu
A replaceable capillary gel electrophoretic (replaceable CGE) method was developed for the separation of two sets of model compounds of single-stranded oligodeoxynucleotide mixtures (18-20 mers), phosphodiester oligodeoxynucleotides (PO-ODNs) and their phosphorothioate modifications (PS-ODNs), with equal sequences differing in a single base. Polyethylene glycol (PEG) 35000 was chosen as the sieving matrix. It was confirmed that PEG polymer solution less influenced resolutions of the PS-ODNs compared with those of the PO-ODNs, while acetonitrile used as an additive in the system improved the separation significantly. It was also noticed that the effect of temperature on separation was much larger than that of denaturant urea. PMID:16809077
Wang, Wenjun; Liao, Bin; Zeng, Ming; Zhu, Chen; Fan, Xianming
Previous study showed that aerosolized signal transducer and activator of transcription-1 (STAT1) antisense oligodeoxynucleotide (ASON) inhibited the expression of STAT1 and ICAM-1 mRNA and protein in alveolar macrophages (AMs) and decreased the concentrations of TGF-β, PDGF and TNF-α in bronchioalveolar lavage fluid (BALF) in bleomycin (BLM)-induced rat pulmonary fibrosis. Administration of STAT1 ASON ameliorated alveolitis in rat pulmonary fibrosis. However, further investigations are neede...
Tamaddon AM.; Hosseini-Shirazi F.; Moghimi HR
Cationic liposomes are used for cellular delivery of antisense oligodeoxynucleotide (AsODN), where release of encapsulated AsODN is mainly controlled by endocytosis and fusion mechanisms. In this investigation, it was tried to model such a release process that is difficult to evaluate in cell culture. For this purpose, an AsODN model (against protein kinase C-α) was encapsulated in a DODAP-containing cationic liposome and evaluated for size, zeta-potential, encapsulation and ODN stab...
Full Text Available Huijie Zhang,1,2 Song Chen,3 Chunyi Zhi,4 Tomohiko Yamazaki,1,2 Nobutaka Hanagata1,2,5 1Graduate School of Life Science, Hokkaido University, Sapporo, Japan; 2Biomaterials Unit, International Center for Materials Nanoarchitectonics, National Institute for Materials Science, Ibaraki, Japan; 3Japanese Society for the Promotion of Science, Tokyo, Japan; 4Department of Physics and Materials Science, City University of Hong Kong, Hong Kong, People’s Republic of China; 5Nanotechnology Innovation Station, Ibaraki, Japan Background: Cytosine-phosphate-guanine (CpG oligodeoxynucleotides activate Toll-like receptor 9, leading to induction of proinflammatory cytokines, which play an important role in induction and maintenance of innate and adaptive immune responses. Previously, we have used boron nitride nanospheres (BNNS as a carrier for delivery of unmodified CpG oligodeoxynucleotides to activate Toll-like receptor 9. However, because CpG oligodeoxynucleotides and BNNS are both negatively charged, electrostatic repulsion between them is likely to reduce the loading of CpG oligodeoxynucleotides onto BNNS. Therefore, the efficiency of uptake of CpG oligodeoxynucleotides is also limited and does not result in induction of a robust cytokine response. To ameliorate these problems, we developed a CpG oligodeoxynucleotide delivery system using chitosan-coated BNNS as a carrier. Methods: To facilitate attachment of CpG oligodeoxynucleotides onto the BNNS and improve their loading capacity, we prepared positively charged BNNS by coating them with chitosan preparations of three different molecular weights and used them as carriers for delivery of CpG oligodeoxynucleotides. Results: The zeta potentials of the BNNS-CS complexes were positive, and chitosan coating improved their dispersity and stability in aqueous solution compared with BNNS. The positive charge of the BNNS-CS complexes greatly improved the loading capacity and cellular uptake efficiency of Cp
... Coast Guard announces the availability of CG-ENG Policy Letter 04-12, ``Alternative Pressure Relief... promulgated regulations on allowable stress factors on May 3, 1979. CG-ENG Policy Letter 04-12 establishes... equivalent to the standards in 46 CFR 154.447 and 46 CFR 154.450. DATES: CG-ENG Policy Letter 04-12...
Strelchenko, Alexei [Fermilab; Stathopoulos, Andreas [William-Mary Coll.
While the incremental eigCG algorithm [ 1 ] is included in many LQCD software packages, its realization on GPU micro-architectures was still missing. In this session we report our experi- ence of the eigCG implementation in the QUDA library. In particular, we will focus on how to employ the mixed precision technique to accelerate solutions of large sparse linear systems with multiple right-hand sides on GPUs. Although application of mixed precision techniques is a well-known optimization approach for linear solvers, its utilization for the eigenvector com- puting within eigCG requires special consideration. We will discuss implementation aspects of the mixed precision deflation and illustrate its numerical behavior on the example of the Wilson twisted mass fermion matrix inversions
Potapova, M V; Nefedova, L N; Kim, A I
The flamenco gene controlling transpositions of the gypsy retrovirus is localized in the 20A1-3 region, in which eight open reading frames organized in a cluster were discovered: DIP1, three repeats of CG32500 and CG32819, and CG14476. Analysis of the genes composing the cluster indicates that their transcription in Drosophila melanogaster is a stage-specific process. Comparison of the expression of these genes in the strains OreR, SS, and MS having the flamenco phenotype and in the strain 413 having the flamenco+ phenotype revealed differences only for the DIP1 gene, transcription of this gene being altered only in the OreR strain. Thus, mutant flamenco alleles are differently expressed in different strains. The structural organization of the flamenco gene region was studied in different Drosophila species: D. sechellia, D. simulans, D. mauritiana, D. yakuba, D. erecta, D. virilis, D. ananassae, D. grimshawi, and D. pseudoobscura. The genes of the cluster were found to be highly conserved in genomes of different species, but in none of them, except D. sechellia, the structural organization of the region repeats the structure of the D. melanogaster cluster. PMID:19947543
Full Text Available Oligodeoxynucleotide (ODN synthesis, which avoids the formation of side products, is of great importance to biochemistry-based technology development. One side reaction of ODN synthesis is the cyanoethylation of the nucleobases. We suppressed this reaction by synthesizing ODNs using fully protected deoxynucleoside 3′-phosphoramidite building blocks, where the remaining reactive nucleobase residues were completely protected with acyl-, diacyl-, and acyl-oxyethylene-type groups. The detailed analysis of cyanoethylation at the nucleobase site showed that N3-protection of the thymine base efficiently suppressed the Michael addition of acrylonitrile. An ODN incorporating N3-cyanoethylthymine was synthesized using the phosphoramidite method, and primer extension reactions involving this ODN template were examined. As a result, the modified thymine produced has been proven to serve as a chain terminator.
Full Text Available Abstract Background The current measure of treatment efficacy of single-dose methotrexate for ectopic pregnancy, is a fall in serum hCG of ≥15% between days 4–7 of treatment, which has a positive predictive value of 93% for treatment success. Two small studies have proposed a fall in serum hCG between days 0–4 after treatment confers similar, earlier prognostic information, with positive predictive values of 100% and 88% for treatment success. We sought to validate this in a large, independent cohort because of the potentially significant clinical implications. Methods We conducted a retrospective study of women (n=206 treated with single-dose methotrexate for ectopic pregnancy (pre-treatment serum hCG levels ≤3000 IU/L at Scottish hospitals between 2006–2011. Women were divided into two cohorts based on whether their serum hCG levels rose or fell between days 0–4 after methotrexate. Treatment outcomes of women in each cohort were compared, and the test performance characteristics calculated. This methodology was repeated for the current measure (≥15% fall in serum hCG between days 4–7 of treatment and an alternate early measure ( Results In our cohort, the positive predictive value of the current clinical measure was 89% (95% CI 84-94% (121/136. A falling serum hCG between days 0–4 predicted treatment success in 85% (95% CI 79-92% of cases (94/110 and a Conclusions We have verified that a decline in serum hCG between days 0–4 after methotrexate treatment for ectopic pregnancies, with pre-treatment serum hCG levels ≤3000 IU/L, provides an early indication of likelihood of treatment success, and performs just as well as the existing measure, which only provides prognostic information on day 7.
Yang, Jiong; Mo, Jingjie; Adamson, Julie T; Håkansson, Kristina
Electron detachment dissociation (EDD), recently introduced by Zubarev and co-workers for the dissociation of multiply charged biomolecular anions via a radical ion intermediate, has been shown to be analogous to electron capture dissociation (ECD) in several respects, including more random peptide fragmentation and retention of labile posttranslational modifications. We have previously demonstrated unique fragmentation behavior in ECD compared to vibrational excitation for oligodeoxynucleotide cations. However, that approach is limited by the poor sensitivity for oligonucleotide ionization in positive ion mode. Here, we show implementation of EDD on a commercial Fourier transform ion cyclotron resonance mass spectrometer utilizing two different configurations: a heated filament electron source and an indirectly heated hollow dispenser cathode electron source. The dispenser cathode configuration provides higher EDD efficiency and additional fragmentation channels for hexamer oligodeoxynucleotides. As in ECD, even-electron d/w ion series dominate the spectra, but we also detect numerous a/z (both even-electron and radical species), (a/z - B), c/x, (c/x - B), and (d/w - B) ions with minimal nucleobase loss from the precursor ions. In contrast to previous high-energy collision-activated dissociation (CAD) and ion trap CAD of radical oligonucleotide anions, we only observe minimum sugar cross-ring cleavage, possibly due to the short time scale of EDD, which limits secondary fragmentation. Thus, EDD provides fragmentation similar to ECD for oligodeoxynucleotides but at enhanced sensitivity. Finally, we show that noncovalent bonding in a DNA duplex can be preserved following EDD, illustrating another analogy with ECD. We believe the latter finding implies EDD has promise for characterization of nucleic acid structure and folding. PMID:15762599
Full Text Available A new photoisomeric nucleoside dUAz bearing an azobenzene group at the C5-position of 2'-deoxyuridine was designed and synthesized. Photoisomerization of dUAz in oligodeoxynucleotides can be achieved rapidly and selectively with 365 nm (forward and 450 nm (backward irradiation. Thermal denaturation experiments revealed that dUAz stabilized the duplex in the cis-form and destabilized it in the trans-form with mismatch discrimination ability comparable to thymidine. These results indicate that dUAz could be a powerful material for reversibly manipulating nucleic acid hybridization with spatiotemporal control.
Lund, O S; Hansen, J E
Oligodeoxynucleotides (ODNs) containing a variable number of 3' and 5' terminal phosphorothioate linkages were applied in free solution to cells infected by HIV-1. ODNs of 28 nt length were applied at up to 5 microM concentration. The ODNs were found to inhibit HIV-1 infection in a dose dependent...... manner, which correlated with the number of modified linkages (4, 8 and 12, respectively). A target sequence in the HIV-1 rev mRNA, previously reported as sensitive to antisense inhibition by full length phosphorothioate ODNs, only revealed non-sequence dependent inhibition of HIV-1, when tested by these...
Ana Rita Rodrigues Barata
Full Text Available El Kathon CG constituye el nombre comercial de una mezcla de isotiazolinas: Metilcloroisotiazolinona y Metilisotiazolinona. Es un conservante muy utilizado, sobre todo en la industria cosmética, aunque en los últimos años también se ha impuesto su presencia en productos de limpieza de uso doméstico y actualmente a concentraciones más altas en preparados de uso industrial como aceites de corte, emulsiones de látex, pinturas al temple, aceites para motores Diesel, etc. Por su alto poder sensibilizante y amplia utilización, constituye actualmente una de las causas más frecuentes de alergia de contacto por preservativos, tanto en nuestra vida privada, como en el ámbito profesional. Objetivos: Estudiar la capacidad sensibilizante del Kathon CG y su relación con el desarrollo de eczema de contacto alérgico de origen profesional. Métodos: Estudio observacional descriptivo, a través de la revisión de las historias clínicas de los pacientes vistos en el Servicio de Dermatología Laboral del Instituto Nacional de Medicina y Seguridad en el Trabajo durante los años 2008-2012; 1520 pacientes fueron evaluados y estudiados mediante pruebas epicutáneas para descartar una posible dermatosis profesional. Se registraron los casos de sensibilización al Kathon CG y se analizaron las siguientes variables: genero, edad, grupo profesional, localización de las lesiones cutáneas, relevancia y relación profesional. El análisis estadístico se realizó con el programa SPSS 15.0. Resultados: Se observó sensibilización al Kathon CG en 88 pacientes (5,8%, correspondiendo un 42% a sensibilizaciones de origen profesional. Conclusiones: La alergia de contacto profesional por Kathon CG constituye actualmente es un problema de alta prevalencia. Ante un paciente con positividad a este alérgeno hay que interrogar siempre por su profesión.Kathon CG is the tradename for a mixture of isothiazolines: methylchloroisothiazolinone and methylisothiazolinone. It
Gilmartin, Daniel J; Soon, Allyson; Thrasivoulou, Christopher; Phillips, Anthony R J; Jayasinghe, Suwan N; Becker, David L
Antisense oligodeoxynucleotides targeting the mRNA of the gap junction protein Cx43 promote tissue repair in a variety of different wounds. Delivery of the antisense drug has most often been achieved by a thermoreversible hydrogel, Pluronic F-127, which is very effective in the short term but does not allow for sustained delivery over several days. For chronic wounds that take a long time to heal, repeated dosing with the drug may be desirable but is not always compatible with conventional treatments such as the weekly changing of compression bandages on venous leg ulcers. Here the coating of collagen scaffolds with antisense oligonucleotides is investigated and a way to provide protection of the oligodeoxynucleotide drug is found in conjunction with sustained release over a 7 d period. This approach significantly reduces the normal foreign body reaction to the scaffold, which induces an increase of Cx43 protein and an inhibition of healing. As a result of the antisense integration into the scaffold, inflammation is reduced with the rate of wound healing and contracture is significantly improved. This coated scaffold approach may be very useful for treating venous leg ulcers and also for providing a sustained release of any other types of oligonucleotide drugs that are being developed. PMID:27253638
Schultz, Kristin N.; Håkansson, Kristina
We have performed extended characterization of the fragmentation patterns of oligodeoxynucleotide dications following electron capture dissociation (ECD) utilizing improved instrumentation. Our current results from a 9.4 T Fourier transform ion cyclotron resonance mass spectrometer equipped with mass-selective external ion accumulation and an indirectly heated dispenser cathode electron source demonstrate much richer fragmentation than from a previous 7 T instrument with a directly heated filament electron source. We propose the previous absence of backbone product ions from purine-containing oligonucleotides is a result of intramolecular hydrogen bonding, preventing product ions from separating. Similar behavior is observed at non-optimized ECD conditions with the current instrumentation. However, infrared laser heating of the resulting charge-reduced radical species results in extensive backbone fragmentation (different from infrared multiphoton dissociation) of the oligonucleotide dA6, demonstrating potential for ECD to characterize nucleic acid secondary structure. Under more optimized conditions, rich fragmentation, mostly in terms of w, (a/z-B), and (c/x-B) products, is observed for dA6, dC6, dG5, and d(GCATGC) following ECD only, allowing complete sequencing in several cases. The current ECD spectra contain some doubly charged products, indicating that populations of gas-phase oligodeoxynucleotide dications are zwitterionic.
The geometry description of radiation shielding codes, such as QAD-CG and MORSE-CG, is often written by the combinatorial geometry (CG) method. However any EGS4/PRESTA user code with the CG method is not yet reported. Since the macro, called $CALL-HOWNEAR, is necessary to use the PRESTA algorithm on EGS4, it is difficult to write complicated geometries on EGS4/PRESTA. Then we developed an EGS4/PRESTA user code, which can be written by 3 dimensional CG method and it did the way that can evaluate influence of the step size for low energy electrons and influence of a calculative geometry. This paper described an user code using the CG method, named PRESTA-CG. Furthermore we reported comparison of the response of an ionization chamber with the UCSAMPCG which is employed the CG description for original EGS4. (author)
H.H. van Ojik; L. Bevaart; C.E. Dahle; A. Bakker (Annie); M.J.H. Jansen (Marco); M.J. van Vugt; J.G. van de Winkel; G.J. Weiner
textabstractImmunostimulatory CpG oligodeoxynucleotides (ODNs) can enhance the therapeutic effect of monoclonal antibodies (mAbs) by enhancing antibody-dependent cell-mediated cytotoxicity (ADCC). Distinct classes of CpG ODNs have been found recently to stimulate different effector
Reiniš, Milan; Šímová, Jana; Bubeník, Jan
Roč. 118, č. 7 (2006), s. 1836-1842. ISSN 0020-7136 R&D Projects: GA ČR(CZ) GA301/04/0492 Institutional research plan: CEZ:AV0Z50520514 Keywords : HPV16 * immunotherapy * CpG oligodeoxynucleotides Subject RIV: EC - Immunology Impact factor: 4.693, year: 2006
Xin, Lusheng; Zhang, Huan; Du, Xinyu; Li, Yiqun; Li, Meijia; Wang, Lingling; Wang, Hao; Qiu, Limei; Song, Linsheng
As a proinflammatory cytokine, vertebrate interleukin 17 (IL17) plays a vital role in the balance of inflammation and homeostasis, and is involved in a systemic regulation of glucose homeostasis. In the present study, a remarkable increase of glucose concentration was observed in oyster serum after 2 d air exposure, which was followed by a rapid up-regulation of CgIL17-1 and CgIL17-5. After oysters was received an injection of extra glucose, the mRNA expressions of CgIL17-1 and CgIL17-5 were also significantly up-regulated. The histopathological changes of hepatopancreas were observed after the oysters were treated by the recombinant proteins of CgIL17-1 and CgIL17-5 in vivo or subjected to air exposure. A significant decrease of GSK3β (Glycogen synthase kinase-3β) protein was also observed after the injection of CgIL17-1 and CgIL17-5 recombinant proteins in vivo. When the oysters with CgIL17-1 and CgIL17-5 genes knocked down were subjected to air exposure, the decline of GSK3β concentration was slowed down and it could still be obviously detected after 7 d compared with that in the control. Meanwhile, the expression of CgDefensin and CgDFFA was inhibited, while CgIAP was up-regulated when CgIL17-1 and CgIL17-5 genes were knocked down, and the oysters exhibited higher mortality (p < 0.05) at 3 d, whereas lower at the late stage of air exposure compared with that in the controls. The results collectively suggested that once oysters were exposed to air, the synthesis of proinflammatory cytokines CgIL17-1 and CgIL17-5 was induced by the up-regulated glucose concentration in oyster serum, which would be not only a negative feedback to the high glucose concentration through mediating the regulation of GSK3β, but also an inducer on tissue damage and immunocompetence as well as the adaptability to stresses. PMID:27268575
Full Text Available Dear Egon!br God bless you on your 95th Birthday! May you complete 100 years.br Being submitted in your honor is a brief article on my continuing work to make available a unique vaccine preventing pregnancy in women without blocking ovulation, her normal production of sex steroid hormones and retaining her regular menstrual cycles and bleeding profiles.br The vaccine is directed at the Human chorionic gonadotropin (hCG, which emerges following fertilization of the egg . Healthy, non-pregnant women do not make it, the basis on which its detection in serum or urine serves as a reliable test for diagnosis of pregnancy. It plays a critical role in implantation of the embryo & thereby to the onset of pregnancy. The purpose of the vaccine is to generate bioeffective antibodies neutralizing hCG & thereby prevent the onset of pregnancy.br As you can imagine, the making of a workable vaccine, competent to make antibodies against a tolerant molecule to the womans immune system (she literally bathes in hCG during pregnancy was not simple. What was also demanded was high immunogenicity of the vaccine to make fairly high titres of antibodies to counteract the large amount of hCG made in early pregnancy. At each stage, it required testing in humans and before that could be done, appropriate toxicology studies & approval of Regulatory and Ethics Committees was needed each taking its time. Eventually the vaccine has to be amenable to industrial production to reach the public, hence a recombinant vaccine had to be developed. Given below is a brief write-up on the evolution of the vaccine against the human chorionic gonadotropin.br Yours, Pran
Background and purpose: Synthetic oligodeoxynucleotides (ODNs) containing unmethylated cytosine-guanine (CpG) motifs bind to Toll-like receptor 9 (TLR9) and stimulate both innate and adaptive immune reactions and possess anti-tumor activity. We recently reported that CpG ODN 1826 strongly enhances radioresponse of both immunogenic [Milas L, Mason K, Ariga H, et al. CpG oligodeoxynucleotide enhances tumor response to radiation. Cancer Res 2004;64:5074-7] and non-immunogenic [Mason KA, Ariga H, Neal R, et al. Targeting toll-like receptor-9 with CpG oligodeoxynucleotides enhances tumor response to fractionated radiotherapy. Clin Cancer Res 2005;11:361-9] murine tumors. Using two immunogenic murine tumors, a fibrosarcoma (FSa) and a mammary carcinoma (MCa-K), the present study explored whether CpG ODN 1826 also improves the response of murine tumors to the chemotherapeutic agent docetaxel (DOC). Materials and methods: CpG ODN 1826 (100 μg) was given sc three times: when leg tumors were 6 mm, when they grew to 8 mm and again 1 week later. DOC (33 mg/kg iv) and local tumor radiation (10 Gy) were given when tumors were 8 mm. Effects of the treatments were assayed by tumor growth delay, defined as days for tumors to grow from 8 to 12 mm in diameter. Results: Treatment with CpG ODN 1826 resulted in strongly enhanced response of FSa tumors to radiation and MCa-K tumors to the chemotherapeutic agent DOC. Enhancement of tumor treatment response was demonstrated by a strong prolongation in the primary tumor treatment endpoint, tumor growth delay. Coincidentally, this treatment also resulted in a higher rate of tumor cure than that observed after tumor radiotherapy or chemotherapy alone. When all three agents were combined the effect was comparable to that of the combination of CpG ODN 1826 with radiation in the case of FSa or of the combination of CpG ODN 1826 with DOC in the case of MCa-K. Conclusion: Overall results show that CpG ODN 1826 can markedly improve tumor response
A double-blind comparative study of CG(L-cysteine) tablets and approved drugs (CG capsules) was made to assess clinical effects of CG tablets on leukopenia. The subjects were 75 patients with cancer of the uterine cervix in whom the number of WBC was 4,500-7,500/mm before irradiation. The ratios of patients who kept WBC as 3,500/mm or more were 64.7% in the group with CG tablets and 50% in the group with CG capsules with no statistical significance. The ratios of patients who kept WBC as 3,000/mm or more were 82.4% in the group with CG tablets and 79.4% in the group with CG capsules with no statistical significance. The ratios to prevent leukopenia were 70.6% in the group with CG tablets and 58.8% in the group with CG capsules with no significant difference. Other hematological findings, subjective symptoms and the performance of radiotherapeutic protocol were not different between the groups. No marked side effects were observed. These results suggest that CG tablets, as well as CG capsules, are effective for preventing leukopenia resulting from radiotherapy. (Namekawa, K.)
Nakashima, Yoshio; Miyagoshi, Takashi; Takamatsu, Mamoru; Sassa, Kazuhiro
In recent years, the color of advertising signboards or vending machines on the streets should be harmonized with the surrounding landscape. In this study, we investigated how the colors (red and white) of the vending machines virtually installed by CG would affect the traditional landscape. 20 subjects estimated landscape samples in Hida-Furukawa by the SD technique. The result of our experiment shows that the vending machines have great influence on the surrounding landscape. On the other hand, we have confirmed that they can harmonize with the circumference landscape by the color to use.
Vollmer, Jörg; Jepsen, Jan Stenvang; Uhlmann, Eugen; Schetter, Christian; Jurk, Marion; Wader, Tanja; Wüllner, Meike; Krieg, Arthur M; Stenvang, Jan
Locked nucleic acid (LNA) is an RNA derivative that when introduced into oligodeoxynucleotides (ODN), mediates high efficacy and stability. CpG ODNs are potent immune stimulators and are recognized by toll-like receptor-9 (TLR9). Some phosphorothioate antisense ODNs bearing CpG dinucleotides have...... been shown to possess immune modulatory capacities. We investigated the effects of LNA substitutions on immune stimulation mediated by antisense ODN G3139 or CpG ODN 2006. LNA ODNs were tested for their ability to stimulate cytokine secretion from human immune cells or TLR9-dependent signaling....... Phosphorothioate chimeric LNA/DNA antisense ODNs with phosphodiester-linked LNA nucleobases at both ends showed a marked decrease of immune modulation with an increasing number of 3' and 5' LNA bases. In addition, guanosine-LNA and cytosine-LNA or simply cytosine-LNA substitutions in the CpG dinucleotides of ODN...
Kamstrup, Søren; Verthelyi, D.; Klinman, D.M.
Exposure to bacterial DNA generates a "danger signal" that stimulates cellular elements of the mammalian immune system to proliferate and/or secrete cytokines. Stimulation is critically dependent on hexameric motifs that contain an unmethylated CpG dinucleotide: these are commonly found in...... bacterial but not vertebrate DNA. Different motifs are optimally stimulatory in different species. This work examines whether oligodeoxynucleotides (ODNs) containing CpG motifs stimulate peripheral blood mononuclear cells from pigs. Results show that pigs respond to CpG ODN by proliferating and secreting IL......-6, IL-12 and TNF-alpha. By screening a large panel (>100) of ODNs, the palindromic hexamer 'ATCGAT' was identified as being optimally active in all animals examined (N = 10). These findings are the first to establish the immunostimulatory activity of CpG ODN in pigs, and suggest that the therapeutic...
Xi-Shan Wang; Kuan Wang; Xue Li; Song-Bin Fu
AIM: To investigate the inhibitory effect of phosphorothioate anti-sense oligodeoxynucleotides (PASODN) on colorectal cancer LS-174T cells in vitro and the mechanism of inhibition of telomerase activity in these cells.METHODS: PASODN were used to infect LS-174T cells and block human telomerase RNA (hTR) through anti-sense technology. The inhibitory effect of PASODN was evaluated by colony-forming inhibition assay and growth curve. Changes of telomerase activity in LS-174T cells were detected by polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA), and the level of apoptosis was analyzed by flow cytometry (FCM) assay.RESULTS: PASODN showed a dose and time-dependent inhibition of cell proliferation. The optimal dosage of PASODN was 10 μmol/L. The colony-forming efficiency was 10.3% in PASODN group after 10 d, whereas that in phosphorothioate mis-sense oligodeoxynucleotides (PMSODN) group with the same concentration and in PBS group (blank control) was 49.1% and 50.7%, respectively. PCR-ELISA results indicated that telomerase activity in the PASODN group was obviously inhibited in comparison with in the control groups (P＜0.01,t = 3.317 and 3.241, t0.01 (20) = 2.845). Meanwhile, before the number of cells was decreased, the morphological changes were observed in the cells of PASODN group. The cells in PASODN group showed the apoptotic peak at 72 h after infection, whereas the control group did not show.CONCLUSION: Specific sequence oligonucleotides can inhibit telomerase activity and lead to cell apoptosis,suggesting a novel treatment strategy for malignant tumors induced by telomerase.
Lijun Chen; Qiuyue Jin; Hong Xie; Ruimin Wang; Li Yao
OBJECTIVE To investigate the effects of survivin antisense oligodeoxynucleotid (ASODN) on proliferation and apoptosis in the chronic myeloid leukemia cell line K562.METHODS Different concentrations of an antisense oligodeoxy-nucleotid and control sequence (scrambled ODN) targeting the survivin gene were transferred into K562 by a lipofectin reagent. The MTT assay was used to measure the growth inhibitory rate, IC50, and to observe the cytotoxicity of survivin ASODN in the K562 cells. The morphologic changes in the nucleus and the apoptotic rate were observed by Hoechst33342/PI staining.Caspase-3 activity was evaluated by a kinase activity assay. The changes of survivin protein expression after transfection were detected by Western blots.RESULTS Eight hours after transfection, fluorescence in the K562 cells was well distributed. Treatment of the cells for 44 h with different concentrations of survivin ASODN produced a IC50 of 800 nmol/L. The growth inhibitory rate with 200, 400, 600 and 1000 nmol/L of survivin ASODN was 15.8±1.6%, 23.8±5.9%, 37.1±5.6% and 77.3±2.5% respectively. After 36 h of of survivin ASODN treatment, distinct morphologic changes characteristic of cell apoptosis such as karyopyknosis and conglomeration were observed by Hoechst33342/PI staining. Caspase-3 activity increased significantly after treatment of the cells with different concentrations of survivin ASODN (P＜0.01)and following treatment with 800 nmol/L survivin ASODN, survivin expression decreased significantly.CONCLUSION Survivin ASODN exerts an anti-cancer effect by inducing apoptosis in K562 leukaemia cells. Up-regulated expression of caspase3 may play a role in this process.
Karla Dias Antunes-Melo
Full Text Available The aims of this study were to improve the cost-benefit ratio of the application of artificial insemination in fixed time (TAI by the transcervical route in sheep, to test the dosage reduction and the use of the vulvar submucosa (VSM route as an alternative for the application of equine chorionic gonadotropin (eCG on the efficiency of the synchronization protocol and fertility to artificial insemination (AI and to measure the level of cortisol in ewes as a result the application of this biotechnique. Blood samples were collected before AI, immediately after AI and seven days after AI. Six groups of twenty animals were used, and each group received doses of 200, 300 and 400 IU of eCG by the intramuscular route (IM and VSM. Estrus was detected, and 48 hours later, the inseminations were performed by the transcervical route. Among the 120 treated ewes, 87.5% came in estrus. The percentage of the intrauterine deposition of semen was 88.3%. The pregnancy rate ranged from 20 to 70% between treatments, with an average of 46.66%. The VSM route was viable for the application of eCG in doses of 300 and 400 IU. The reduction in the eCG dose to 200 IU by the IM route reduces costs while maintaining the efficiency of estrus synchronization protocols and TAI with frozen semen in Santa Ines ewes. The average levels of cortisol were significant, at 1.15, 2.86 and 0.52 ?g/dL according to the three collections, being higher after the animal was off the easel at the end of AI. The procedures for performing the transcervical AI technique indicate that stress in the animals produces satisfactory fertility results and a low cost in multiparous ewes of the Santa Ines breed.
Karla Dias Antunes-Melo; Veronaldo Souza de Oliveira; Anselmo Domingos Ferreira Santos; Claudio Alvarenga de Oliveira; Lícia Mendes Mendonça; Julianne Santiago Silva Goveia; Thiago Santos Almeida
The aims of this study were to improve the cost-benefit ratio of the application of artificial insemination in fixed time (TAI) by the transcervical route in sheep, to test the dosage reduction and the use of the vulvar submucosa (VSM) route as an alternative for the application of equine chorionic gonadotropin (eCG) on the efficiency of the synchronization protocol and fertility to artificial insemination (AI) and to measure the level of cortisol in ewes as a result the application of this b...
国华; 梁华平; 吕凤林; 徐祥; 安兵; 郝天智; 王付龙; 杨文军
Objective: To study the effect of decoy-oligodeoxynucleotides (decoy-ODNs) in dumbbell shape with the oligodeoxynucleotide sequence similar to nuclear factor kappa B (NF-κB) cis-elements on expression of inflammation mediators in pMΦ cells from rats.Methods: With carriers of cationic liposomes, decoy-ODNs were transfected into pMΦ cells of rats. Then the inhibiting effects of the decoy-ODNs on tumor necrosis factor α (TNFα), interleukin-6 (IL-6) and IL-10 were analyzed.Results: Decoy-ODNs could decrease the expression of TNFα and IL-6 in dose-dependent fashion but had weaker inhibiting effect on IL-10. Conclusions: Decoy-ODNs targeting NF-κB can decrease the expression of inflammatory mediators in pMΦ cells from rats.
Objective: To investigate whether bcl-2 antisense oligodeoxynucleotide (ASODN) could enhance 60Co γ-ray induced-apoptosis in malignant lymphoma cells. Methods: Cell apoptosis situation, cellular DNA contents and bcl-2 protein expression were determined by cell stain, TdT-mediated dUTP Nick-end Labeling (TUNEL) and flow cytometry. Results: One to 8 Gy γ-ray and 10-40 μmol/L bcl-2 ASODN could inhibit Raji cell growth and induce cell apoptosis. Assay of flow cytometry showed that the number of Sub-G1 cells was significantly increased and Bcl-2 protein expression in Raji cells was significantly downregulated, which showed time- and dose-dependent manners. The effects were more pronounced when γ-ray and bcl-2 ASODN were used together rather than either alone. Conclusion: Bcl-2 antisense oligodeoxynucleotide could enhance 60Co γ-ray induced-apoptosis in malignant lymphoma cells. (authors)
GONGHai-Xia; GUOXi-Rong; FEILi; GUOMei; LIUQian-Qi; CHENRong-Hua
AIM:To investigate the influence of central administration of neuropeptide Y-Y5 receptor antisense oligodeoxynucleotides(ODN) on the body weight and fat pads of high-energy diet-induced obese rats, and the effects on white adipocyte lipolysis and apoptosis. METHODS: Y5 receptor antisense, sense, mismatched oligodeoxynucleotides (ODN) or vehicle were intracerebroventricularly injected, and average adipocyte area was calculated. DNA ladders were measured to evaluate adipocyte apoptosis, and RT-PCR was used to analyze the expression of bcl-2 and bax gene. RESULTS: (1) Central administration of Y5 receptor antisense ODN significantly decreased body weight, fat pads, and average adipocyte area. (2) DNA fragmentation was presented after electrophoresis at both epididymal and retroperitoneal adipose tissue. (3) The expression of bcl-2 gene was downregulated, while the expression of bax was upregulated. CONCLUSION:Lipolysis and adipocyte apoptosis may be important reasons for Y5 receptor antisense therapy.
Hendrix, C.; Devreese, B.; Rozenski, J; Van Aerschot, A; Bruyn, A.; Van Beeumen, J.; Herdewijn, P
The functionalisation of oligodeoxynucleotides and oligoribonucleotides by incorporation of 2'-amido-2'-deoxyribonucleosides, possibly containing a reporter group via the 2'-amido bond, was examined. Therefore 2'-acetamido-ribonucleosides containing a small methyl group at the 2'-amido bond were synthesized as model compounds. In order to evaluate the influence of this 2'-modification on the hybridization capacities, 2'-acetamido-2'-deoxyuridine was incorporated in both RNA and DNA strands. T...
Bates, P. J.; Dosanjh, H. S.; S. Kumar; Jenkins, T. C.; Laughton, C A; Neidle, S
Real-time biomolecular interaction analysis (BIA) has been applied to triplex formation between oligodeoxynucleotides. 5'-Biotinylated oligonucleotides were immobilised on the streptavidin-coated surface of a biosensor chip and subsequently hybridised to their complementary strand. Sequence-specific triplex formation was observed when a suitable third-strand oligopyrimidine was injected over the surface-bound duplex. In addition, a single-stranded oligonucleotide immobilised on the chip surfa...
Ratajczak, M Z; Kant, J A; Luger, S M; Hijiya, N.; Zhang, J; Zon, G; Gewirtz, A. M.
The c-myb protooncogene encodes proteins that are critical for hematopoietic cell proliferation and development. Disrupting c-myb function might, therefore, prove an effective therapeutic strategy for controlling leukemic cell growth. Antisense oligodeoxynucleotides have been utilized for this purpose in vitro, but their in vivo efficacy has not been reported. We therefore established human leukemia-scid mouse chimeras with K562 cells and treated diseased animals with phosphorothioate-modifie...
Onizuka, Kazumitsu; Taniguchi, Yosuke; Sasaki, Shigeki
Site-specific modification of RNA is of great significance to investigate RNA structure, function and dynamics. Recently, we reported a new method for sequence- and cytosine-selective chemical modification of RNA based on the functional group transfer reaction of the 1-phenyl-2-methylydene-1,3-diketone unit of the 6-thioguanosine base incorporated in the oligodeoxynucleotide probe. In this study, we describe that the functionality transfer rate is greatly enhanced and the selectivity is shift...
Parr, Carol; Pierce, Sarah E.; Smith, Suncerae I.; Brodbelt, Jennifer S.
The reactions of two well-known chemical probes, glyoxal and potassium permanganate (KMnO4), with oligodeoxynucleotides were monitored by electrospray ionization (ESI) mass spectrometry to evaluate the influence of the sequence of DNA, its secondary structure, and interactions with associated ligands on the reactivity of the two probes. Glyoxal, a guanine-reactive probe, incorporated a mass shift of 58 Da, and potassium permanganate (KMnO4) is a thymine-reactive probe that resulted in a mass ...
Highlights: ► Ni–SiC–Cg nanocomposite coating was deposited onto the St-37 substrate by electrodeposition method. ► From XRD, all of the coatings show high crystallinity degree. ► All the samples had a typical cluster like structures with the average crystallites size of 60 nm. ► Mechanical properties were improved with increasing of pH values up to 4.8 (593 HV). ► At pH equal to 4.8, Ni–SiC–Cg nanocomposite coating with suitable characteristics has been obtained. - Abstract: Nickel–silicon carbide–graphite (Ni–SiC–Cg) nanocomposite coating was deposited onto the steel (St-37) substrate by electrochemical plating method. The influence of pH parameter on the mechanical properties and morphological features was studied. The coatings were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), Elemental mapping analysis system, and field emission scanning electron microscopy (FE-SEM) techniques. Besides, the microhardness of specimens was measured by Micro hardness Tester. Based on XRD results, the dominant phases with high crystallinity degree were nickel, silicon carbide and graphite. Microscopic observations illustrated that all the coatings had a cluster like structures which consisted of some fine sphere particles with an average crystallites size of 60 nm. Among of all coatings, the specimen at pH equal to 4.8 presented an appropriate morphology as well as lower porosity with high grain boundaries. Moreover, elemental mapping spectra exhibited a homogenous distribution of nickel, silicon and carbon particles. However, the distribution of silicon particles into nickel matrix at pH = 4.8 was higher than the other samples. Also, mechanical properties of the attained coatings were improved with increasing of pH values up to 4.8 (593 HV) which is in agreement with microstructural characteristics. Therefore, obtaining of the optimum pH value is the major parameter in
Sendag, Sait; Cetin, Yunus; Alan, Muhammet; Hadeler, Klaus-Gerd; Niemann, Heiner
The goal of the present study was to compare the ovarian response, oocyte yields per animal, and the morphological quality of oocytes collected by ultrasound guided follicular aspiration from Holstein cows treated either with FSH or eCG. Twenty four normal cyclic, German Holstein cows were randomly divided into two groups. Fourteen cows received 3000 IU eCG on day-4 prior to ovum pick-up (OPU) (day 0), 2 days later (day-2), 625 microg cloprostenol was administered. On day-1 GnRH was administered i.m. and 24h later OPU (day 0) was performed. In ten cows a total dose of 500 IU follicle stimulating hormone (Pluset) was administered intramuscularly in a constant dosage for 4 days with intervals of 12h, starting on day-5. Luteolysis was induced by application of 625 microg cloprostenol on day-2. On day-1 (24h after the last FSH treatment) GnRH was administered i.m. and 24h later OPU (day 0) was performed. Ovarian follicles were visualized on the ultrasound monitor, counted and recorded. All visible antral follicles were punctured. Recovered oocytes were graded morphologically based on the cumulus investment. Average follicle number in ovaries was higher in FSH group than eCG group (pgrade A oocytes was higher in the FSH group in the than eCG group (pgrade C oocytes in FSH group were lower than eCG group (povaries and oocyte quality are affected by the type of gonadotropin and FSH is better alternative than eCG for OPU treatment. PMID:18294785
Full Text Available The major weaknesses of subunit vaccines are their low immunogenicity and poor efficacy. Adjuvants can help to overcome some of these inherent defects with subunit vaccines. Here, we evaluated the efficacy of the newly developed water-in-oil-in-water multiphase emulsion system, termed PELC, in potentiating the protective capacity of dengue-1 envelope protein domain III. Unlike aluminum phosphate, dengue-1 envelope protein domain III formulated with PELC plus CpG oligodeoxynucleotides induced neutralizing antibodies against dengue-1 virus and increased the splenocyte secretion of IFN-γ after in vitro re-stimulation. The induced antibodies contained both the IgG1 and IgG2a subclasses. A rapid anamnestic neutralizing antibody response against a live dengue virus challenge was elicited at week 26 after the first immunization. These results demonstrate that PELC plus CpG oligodeoxynucleotides broaden the dengue-1 envelope protein domain III-specific immune responses. PELC plus CpG oligodeoxynucleotides is a promising adjuvant for recombinant protein based vaccination against dengue virus.
Fan YANG; Zu-mei SHI; Hui JIN; Li-ping ZHU; Kun-ming LI; Jian-zhi YANG; Zhi-qin CHEN; Xiao-ming TENG; Hui-fen CHEN; Yu WANG
Objective To compare the different time of administration of hCG affecting pregnancy rate of IUI. Methods A total of 189 infertile couples underwent 331 cycles of IUI with husband’s sperm.They were separated into 3 groups according to the time of hCG administration in IUI:hCG 1-23 h(group A):hCG 24-36 h (group B);hCG 37—48 h(group C). Results There were no statistical differences among 3 groups.None of the other relative factors，such as the female age，the different methods of ovulation and the cause of infertility,showed differences in pregnancy rate among 3 groups. Conclusion IUI can be performed any time after administration of hCG(1—48 h).
Bollwein, H; Braun, J
In this study the use of hCG for induction of ovulation is described. Factors such as follicle diameter at the time of administration of hCG (3000 IE hCG i.v.), follicular growth after hCG and the rate of double ovulations were evaluated. A total of 168 mares presented for artificial insemination were used. In 249 estrous periods hCG was given to mares exhibiting standing estrous when a minimum follicle diameter of 30 mm and a well developed edema of the endometrium could be detected by ultrasonography. In nine estrous periods ovulation occurred within 24 hours after hCG. The majority of mares (216; 86.7%) ovulated 24 to 48 hours after hCG and in 24 cases ovulation was delayed beyond 48 hours. Follicle size at the time of hCG administration (30-34 mm, 35-39 mm, > or = 40 mm) had no influence on the percentage of mares ovulating 24 to 48 hours after hCG (89.2%, 87.9%, and 83.7%, respectively). Double ovulations could be observed in 17.7% of estrous periods. The one cycle pregnancy rate was not influenced by follicle size (small 45.9%; medium 41.6%; large 47.5%). Repeated treatments with hCG during successive estrous cycles within one year did not influence the rate of responding to hCG. Mares in standing estrous respond well to hCG if a minimum follicle size of 30 mm and a well developed endometrial folding is present. PMID:10077815
To study the effect of c-myc antisense oligodeoxynucleotides (ODNs) on proliferation of pulmonary vascular pericytes (PC) induced by hypoxia, cell culture, dot hybridization using probe of digoxigenin-11-dUTP-labeled cDNA,3H-thymidine incorporation, immunocytochemical technique and image analysis methods were used to observe the effect of c-myc antisense ODNs on expression of c-myc gene and proliferating cell nuclear antigen (PCNA), and 3H-thymidine incorporation of PC induced by hypoxia. The results showed that hypoxia could significantly enhance the expression of c-myc and PCNA (P＜0.01), and elevate 3H-thymidine incorporation of PC (P＜0.01), but antisense ODNs could significantly inhibit the expression of c-myc and PCNA (P＜0.05), and 3H-thymidine incorporation of PC (P＜0.01). It was suggested that hypoxia could promote the proliferation of PC by up-regulating the expression of c-myc gene, but c-myc antisense ODNs could inhibit hypoxia-induced proliferation of PC by downregulating the expression of c-myc gene.
Betting, David J; Hurvitz, Sara A; Steward, Kristopher K; Yamada, Reiko E; Kafi, Kamran; van Rooijen, Nico; Timmerman, John M
Rituximab plus chemotherapy is standard therapy for patients with non-Hodgkin B cell lymphoma, but often complete response or cure is not achieved. Toll-like receptor 9 agonist CpG oligodeoxynucleotides (CpG) can improve antibody-dependent cellular cytotoxicity and adaptive antitumor immune responses. Using a syngeneic murine B cell lymphoma expressing human CD20 (38C13-huCD20), we previously demonstrated that rituximab plus intratumoral CpG, but not systemic CpG, could eradicate up to half of 7-day established 38C13-huCD20 tumors. However, larger 10-day established tumors could not be cured with this regimen. We thus hypothesized that cytoreduction with cyclophosphamide (Cy) before immunotherapy might permit eradication of these more advanced tumor burdens. Pretreatment with Cy resulted in tumor eradication from 83% of animals treated with rituximab/CpG, whereas Cy/CpG or Cy/rituximab treatments only cured 30% or 17%, respectively (P<0.005). Tumor eradication depended on natural killer cells, but not T cells, macrophages, or complement. Only mice treated with Cy/rituximab/CpG partially resisted rechallenge with tumor cells. Foxp3 Treg and CD11bGr1 myeloid suppressor cells persisted within lymphoid organs after therapy, possibly influencing the ability to establish adaptive tumor immunity. In conclusion, cytoreduction with Cy permitted the cure of large, established lymphomas not otherwise responsive to rituximab plus intratumoral CpG immunotherapy. PMID:22892450
Highlights: • A shRNA vector based transcription factor decoy, VB-ODN, was designed. • VB-ODN for NF-κB inhibited cell viability in HEK293 cells. • VB-ODN inhibited expression of downstream genes of target transcription factors. • VB-ODN may enhance nuclear entry ratio for its feasibility of virus production. - Abstract: In this study, we designed a short hairpin RNA vector-based oligodeoxynucleotide (VB-ODN) carrying transcription factor (TF) consensus sequence which could function as a decoy to block TF activity. Specifically, VB-ODN for Nuclear factor-κB (NF-κB) could inhibit cell viability and decrease downstream gene expression in HEK293 cells without affecting expression of NF-κB itself. The specific binding between VB-ODN produced double-stranded RNA and NF-κB was evidenced by electrophoretic mobility shift assay. Moreover, similar VB-ODNs designed for three other TFs also inhibit their downstream gene expression but not that of themselves. Our study provides a new design of decoy for blocking TF activity
Oligodeoxynucleotide sequences that uniquely complemented 16S rRNAs of each group of methylotrophs were synthesized and used as hybridization probes for the identification of methylotrophic bacteria possessing the serine and ribulose monophosphate (RuMP) pathways for formaldehyde fixation. The specificity of the probes was determined by hybridizing radiolabeled probes with slot-blotted RNAs of methylotrophs and other eubacteria followed by autoradiography. The washing temperature was determined experimentally to be 50 and 52 degrees C for 9-α (serine pathway) and 10-γ (RuMP pathway) probes, respectively. RNAs isolated from serine pathway methylotrophs bound to probe 9-α, and RNAs from RuMP pathway methylotrophs bound to probe 10-γ. Nonmethylotrophic eubacterial RNAs did not bind to either probe. The probes were also labeled with fluorescent dyes. Cells fixed to microscope slides were hybridized with these probes, washed, and examined in a fluorescence microscope equipped with appropriate filter sets. Cells of methylotrophic bacteria possessing the serine or RuMP pathway specifically bind probes designed for each group. Samples with a mixture of cells of type I and II methanotrophs were detected and differentiated with single probes or mixed probes labeled with different fluorescent dyes, which enabled the detection of both types of cells in the same microscopic field
Full Text Available To compare the efficiency of novel mitogenic agents and traditional mitosis inductors, 18 patients with splenic marginal zone lymphoma (SMZL were studied. Three cultures using oligodeoxynucleotide (ODN plus interleukin-2 (IL-2, or TPA, or LPS were setup in each patient. Seventeen/18 cases with ODN+IL2 had moderate/good proliferation (94,4% as compared with 10/18 cases with TPA and LPS (55% (P=.015; 14/18 (77,7% cases with ODN+IL2 had sufficient good quality of banding as compared with 8/18 cases (44,4% with TPA and LPS. The karyotype could be defined from ODN+IL2-stimulated cultures in all 18 patients, 14 of whom (77,7% had a cytogenetic aberration, whereas clonal aberrations could be documented in 9 and in 3 cases by stimulation with LPS and TPA, respectively. Recurrent chromosome aberrations in our series were represented by aberrations of chromosome 14q in 5 patients, by trisomy 12 and 7q deletion in 4 cases each, and by abnormalities involving 11q and 13q in two cases each. These findings show that stimulation with ODN+IL2 offers more mitotic figures of better quality and results in an increased rate of clonal aberrations in SMZL, making this method ideal for prospective studies aiming at the definition of the prognostic impact of cytogenetic aberrations in this disorder.
Full Text Available The paper explored the regulatory role of oligodeoxynucleotides (ODNs with specific sequences in the proliferation and activation of osteoblast, using human osteoblast-like cell line MG 63 as the model. Through the administration of ODNs to MG 63 cells at a concentration of 1.0 µg/mL, ODN MT01 with positive effects on proliferation and activation of osteoblast was selected from 11 different ODNs by methyl thiazolyl tetrazolium (MTT assay and alkaline phosphatase (ALP activity measurement. To get a deeper insight into the molecular mechanism, effects of ODN MT01 treatment on the expression level of Sp7, runx-2, collagen-I, osteoprotegerin (OPG and RANK ligand (RANKL were determined using quantitative real time PCR and Western blotting. Remarkably, the mRNA and protein expression levels of Sp7, runx-2, collagen-I and OPG were improved after ODN MT01 treatment. Meanwhile, the protein expression level of RANKL was dramatically decreased. These results suggested that ODN MT01 had a significant impact in facilitating osteogenic proliferation and activation, and provided a direct evidence for the notion that single strand ODN could regulate the balance of bone formation and resorption, and thus was of great potential in the rebuilding of alveolar bone.
Murad, Yanal M; Clay, Timothy M
Toll-like receptors (TLRs) are part of the innate immune system, and they belong to the pattern recognition receptors (PRR) family. The PRR family is designed to recognize and bind conserved pathogen-associated molecular patterns, which are not generated by the host and are restricted and essential to micro-organisms. TLR9, which recognizes unmethylated CpG (cytosine guanosine dinucleotide), is a very promising target for therapeutic activation. Stimulation of TLR9 activates human plasmacytoid dendritic cells and B cells, and results in potent T helper-1 (T(h)1)-type immune responses and antitumor responses in mouse tumor models and in patients. Several pharmaceutical companies, such as Pfizer, Idera, and Dynavax, are developing CpG oligodeoxynucleotides (ODNs) for the treatment of cancer, along with other conditions, such as infections and allergy. CpG ODNs have shown promising results as vaccine adjuvants and in combination with cancer immunotherapy. Several TLR9 agonists are being developed and have entered clinical trials to evaluate their safety and efficacy for the treatment of several hematopoietic and solid tumors. In this review, we discuss the use of CpG ODNs in several phase I and II clinical trials for the treatment of NHL, renal cell carcinoma, melanoma, and non-small cell lung cancer, either alone or in combination with other agents. PMID:19894778
Xiao, Xiao [State Key Laboratory of Cancer Biology and Xijing Hospital of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi Province (China); Gang, Yi [State Key Laboratory of Cancer Biology and Xijing Hospital of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi Province (China); Department of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xi’an 710038, Shaanxi Province (China); Wang, Honghong [No. 518 Hospital of Chinese People’s Liberation Army, Xi’an 710043, Shaanxi Province (China); Wang, Jiayin [The Genome Institute, Washington University in St. Louis, St. Louis, MO 63108 (United States); Zhao, Lina [Department of Radiation Oncology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi Province (China); Xu, Li, E-mail: email@example.com [State Key Laboratory of Cancer Biology and Xijing Hospital of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi Province (China); Liu, Zhiguo, E-mail: firstname.lastname@example.org [State Key Laboratory of Cancer Biology and Xijing Hospital of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi Province (China)
Highlights: • A shRNA vector based transcription factor decoy, VB-ODN, was designed. • VB-ODN for NF-κB inhibited cell viability in HEK293 cells. • VB-ODN inhibited expression of downstream genes of target transcription factors. • VB-ODN may enhance nuclear entry ratio for its feasibility of virus production. - Abstract: In this study, we designed a short hairpin RNA vector-based oligodeoxynucleotide (VB-ODN) carrying transcription factor (TF) consensus sequence which could function as a decoy to block TF activity. Specifically, VB-ODN for Nuclear factor-κB (NF-κB) could inhibit cell viability and decrease downstream gene expression in HEK293 cells without affecting expression of NF-κB itself. The specific binding between VB-ODN produced double-stranded RNA and NF-κB was evidenced by electrophoretic mobility shift assay. Moreover, similar VB-ODNs designed for three other TFs also inhibit their downstream gene expression but not that of themselves. Our study provides a new design of decoy for blocking TF activity.
Sethi, Shneh; Thormann, Ulrich; Sommer, Ursula; Stötzel, Sabine; Mohamed, Walid; Schnettler, Reinhard; Domann, Eugen; Chakraborty, Trinad; Alt, Volker
TLR-9 ligand CpG oligodeoxynucleotide type B (CpG ODN) induces a proinflammatory environment. We evaluated the effects of a preoperative CpG ODN application in an implant-associated Staphylococcus aureus bone infection model by monitoring bacterial loads and cytokine and chemokine levels. A total of 95 rats were used in four different groups: CpG ODN group (group 1; n=25), non-CpG-ODN group (group 2; n=25); saline pretreatment (group 3; n=25), and one uninfected group (group 4; n=20). A single dose of CpG-ODN was administered to the left tibialis anterior muscle 3days prior to surgery and the tibia midshaft was osteotomized, stabilized by an intramedullary implant and subsequently contaminated with 10(3) colony forming units (CFUs) of S. aureus in groups 1-3. The osteotomy gap in animals of group 4 was not contaminated with S. aureus and those animals did not receive any pretreatment. CpG ODN administration resulted in significant reduction of the bacterial load in tibia tissue homogenate and on the implant surface on day 1 post-infection compared to non-CpG-ODN pretreatment (pmodel, prophylactic administration of a single dose of CpG ODN, resulted in marked reduction of S. aureus load in the infected tibia during the initial stage of infection but failed to prevent development of chronic infection over time. PMID:25959416
Zhang, Xiaoling; Sun, Wei; Wu, Xiuli; Wang, Hua; Yan, Youyou; Guo, Sheng; Song, Dandan; Li, Hainan; Gao, Shuang; Wang, Luowei; Yu, Yongli; Wang, Liying
Toll-like receptor 9 (TLR9) can sense pathogen DNA and CpG ODN or even self-DNA by trafficking assisted by Unc93B1, an endoplasmic reticulum (ER) transmembrane protein, from ER to endolysosomes or cell surface. In previous study, we found that an oligodeoxynucleotide with CCT repeats (SAT05f) could selectively inhibit TLR7/9 activation. However, the mechanism for the inhibitory activity of SAT05f is still unknown. In present research, it was found that SAT05f could inhibit CpG ODN-induced the intracellular trafficking of TLR9 and Unc93B1 with feedback the responses of decreased surface TLR9 and enhanced TLR9 mRNA expression but not influence TLR9 protein level by using human plasmacytoid dendritic cell line CAL-1 cells, suggesting that SAT05f inhibits TLR9 activation by restraining TLR9 trafficking. Since the mitochondrial DNA released from injured tissue can cause systemic inflammatory response syndrome (SIRS), this study may provide valuable data for prevention and treatment of SIRS and rescue severe trauma patients. PMID:25374030
Nagpal, Gandharva; Gupta, Sudheer; Chaudhary, Kumardeep; Dhanda, Sandeep Kumar; Prakash, Satya; Raghava, Gajendra P S
Immunomodulatory oligodeoxynucleotides (IMODNs) are the short DNA sequences that activate the innate immune system via toll-like receptor 9. These sequences predominantly contain unmethylated CpG motifs. In this work, we describe VaccineDA (Vaccine DNA adjuvants), a web-based resource developed to design IMODN-based vaccine adjuvants. We collected and analyzed 2193 experimentally validated IMODNs obtained from the literature. Certain types of nucleotides (e.g., T, GT, TC, TT, CGT, TCG, TTT) are dominant in IMODNs. Based on these observations, we developed support vector machine-based models to predict IMODNs using various compositions. The developed models achieved the maximum Matthews Correlation Coefficient (MCC) of 0.75 with an accuracy of 87.57% using the pentanucleotide composition. The integration of motif information further improved the performance of our model from the MCC of 0.75 to 0.77. Similarly, models were developed to predict palindromic IMODNs and attained a maximum MCC of 0.84 with the accuracy of 91.94%. These models were evaluated using a five-fold cross-validation technique as well as validated on an independent dataset. The models developed in this study were integrated into VaccineDA to provide a wide range of services that facilitate the design of DNA-based vaccine adjuvants (http://crdd.osdd.net/raghava/vaccineda/). PMID:26212482
Purpose: To investigate whether irradiation before antisense Bcl-2 oligodeoxynucleotide (ODN) administration enhances tissue uptake, and whether periodic dosing enhances cellular uptake of fluorescently labeled ODN relative to constant dosing. Methods and Materials: PC-3-Bcl-2 cells (prostate cancer cell line engineered to overexpress Bcl-2) were subjected to increasing doses of irradiation (0-10 Gy) with or without increasing concentrations of fluorescently labeled antisense Bcl-2 ODN (G4243). The fluorescent signal intensity was quantified as the total grain area with commercial software. In addition, PC-3-Bcl-2 subcutaneous xenograft tumors were treated with or without irradiation in combination with various dosing schemas of G4243. The uptake of fluorescent G4243 in tumors was quantitated. Results: The uptake of G4243 was increased in prostate cancer cells exposed to low doses of irradiation both in vitro and in vivo. Irradiation before G4243 treatment resulted in increased fluorescent signal intensity in xenograft tumors compared with those irradiated after G4243 treatment. A single weekly dose of G4243 produced higher G4243 uptake in xenograft tumors than daily dosing, even when the total dose administered per week was held constant. Conclusions: These findings suggest that ionizing radiation increases the uptake of therapeutic ODN in target tissues and, thus, has potential to increase the efficacy of ODN in clinical applications
YANGPeng-Yuan; RUIYao-Cheng; JINYou-Xin; LITie-Jun; QIUYan; ZHANGLi; WANGJie-Song
AIM:To study the expression of vascular endothelial growth factor (VEGF) induced by oxidized low density liprotein (ox-LDL) and the inhibitory effects of antisense oligodeoxynucleotide (asODN) on the levels of VEGF protein and mRNA in the U937 foam cells. METHODS: U937 cells were incubated with ox-LDL 80 mg/L for 48h, then ,the foam cells were treated with asODN (0,5,10, and 20μmol/L). The VEGF concentration in the media was determined by ELISA. The VEGF protein expression level in cells was measured by immuohistochemistry; the positive ratio detected by a morphometrical analysis system was used as the amount of the VEGF expression level. The VEGF mRNA level was examined by Northern blotting. RESULTS: After U937 cells were incubated with ox-LDL, VEGF expression level increased greatly both in the cells and in the media. asODN markeldy inhibited the increase of VEGF. After treatment with asODN 20μmol/L, the VEGF protein concentration in the media decreased by 45.0%， the VEGF positive ratio detected by immuohistochemistry in cells decreased by 64.9%, and the VEGF mRNA level decreased by 47.1%. CONCLUSION: The expression of VEGF in U937 foam cells was strong. asODN inhibited VEGF expression significantly in U937 foam cells in vitro.
Full Text Available CpG-oligodeoxynucleotides (CpG-ODN are potent immune stimuli being developed for use as adjuvants in different species. Toll-like receptor 9 (TLR9 is the cellular receptor for CpG-ODN in mammalian cells. The CpG-ODN with 18-24 deoxynucleotides that are in current use for human and mouse cells, however, have low activity with rabbit TLR9. Using a cell-based activation assay, we developed a type of CpG-ODN containing a GACGTT or AACGTT motif in 12 phosphorothioate-modified deoxynucleotides with potent stimulatory activity for rabbit TLR9. The developed CpG-ODN have higher activities than other developed CpG-ODN in eliciting antigen-nonspecific immune responses in rabbit splenocytes. When mixed with an NJ85 peptide derived from rabbit hemorrhagic disease virus, they had potent activities to boost an antigen-specific T cell activation and antibody production in rabbits. Compared to Freund's adjuvant, the developed CpG-ODN are capable of boosting a potent and less toxic antibody response. The results of this study suggest that both the choice of CpG-motif and its length are important factors for CpG-ODN to effectively activate rabbit TLR9 mediated immune responses.
杜晓岩; 王健; 徐万祥; 朱康杰; 蒋宏亮; 刘世范
This paper describes the study of the immunological response of C57 Black mouse to hCG by multiple inoculations with hCG and single injection with a long-term release system of hCG/poly (lactide co-glycolide ) microsphere (hCG/PGLA ). Three parameters including 1) the dynamic variation of hCG antibody titres , 2) the capacity of biological neutralization by hCG antibodies in vitro and in vivo, and 3 ) assay of the binding affinity constants of hCG antibodies, are compared. The results showed that the single injection with hCG/PGLA was capable of producing high titres of anti-hCG antibodies. The analyses of three parameters of antisera from C57 Black mouse shozoed that the properties of the antisera elicited using a single injection with hCG/PGLA were generally similar to those of the antisera raised by the multiple inoculations with hCG, and concentration of the antibodies was directly proportional to hCG bioneutralization capacity. Meanwhile, all of these results suggest us that a further long-term release system of β-hCG/PGLA as a vaccine may substitute for conventional vaccine and it also provides a new convenient inoculative method for widespread clinical application and research of hCG vaccine and other vaccines.
T.I.M. Korevaar (Tim); E.A.P. Steegers (Eric); Y.B. de Rijke (Yolanda); S. Schalekamp-Timmermans (S.); W. Visser (Willy); A. Hofman (Albert); V.W.V. Jaddoe (Vincent); H.W. Tiemeier (Henning); T.J. Visser (Theo); M. Medici (Marco); R.P. Peeters (Robin)
textabstractHuman chorionic gonadotropin (hCG) is a pregnancy hormone secreted by the placental synctiotrophoblast cell layer that has been linked to fetal growth and various placental, uterine and fetal functions. In order to investigate the effects of hCG on clinical endpoints, knowledge on refere
Risselada, Arne; Vehof, Jelle; Bruggeman, Richard; Wilffert, Bob; Cohen, Dan; Al Hadithy, Asmar F.; Arends, Johan; Mulder, Hans
Background: Studies have found an association between the ADRA2A 1291 C/G polymorphism and antipsychoticinduced weight gain. A possible association with the metabolic syndrome has not been studied. Objectives: To investigate the association between the ADRA2A 1291 C/G polymorphism and the metabolic
... Security Measures Summary (Form CG-6025) A Appendix A to Part 105 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY MARITIME SECURITY MARITIME SECURITY: FACILITIES Pt. 105, App. A Appendix A to Part 105—Facility Vulnerability and Security Measures Summary (Form CG-6025)...
Full Text Available Abstract Background In natural cycles, women conceive when intercourse takes place during a six-day period ending on the day of ovulation. The current practice in intrauterine insemination (IUI cycles is to perform the IUI 24-36 hours after the hCG administration, when the ovulation is already imminent. In this study hCG was administered after the IUI, which more closely resembles the fertilisation process in natural cycles. Methods All the IUIs performed since the beginning of 2007 were analysed retrospectively. Our standard protocol has been to perform the IUI 24-32 hours after hCG administration. From the end of 2008, we started to inject hCG after the IUI at random. The main outcome measure was the result of a urinary pregnancy test. Generalized Estimating Equations (GEE was used to identify independent factors affecting the cycle outcome. Results The analysis included 228 cycles with hCG administered before and 104 cycles hCG administered after the IUI. The pregnancy rates were 10.9% and 19.6% (P = 0.040, respectively. Independent factors (OR, 95% CI affecting the cycle outcome were sperm count (2.65, 1.20-5.81, number of follicles > 16 mm at IUI (2.01, 1.07-3.81 and the time of hCG administration (2.21, 1.16-4.19. Conclusion Improved pregnancy rate was observed with administration of hCG after IUI.
Santerre, Christelle; Sourdaine, Pascal; Adeline, Béatrice; Martinez, Anne-Sophie
Sex determination is poorly known in mollusks, lophotrochozoa and most hermaphrodites. In the oyster Crassostrea gigas, an irregular successive hermaphrodite, sex determination may occur at the end of a gametogenetic cycle to prepare the next cycle. To investigate further into these aspects we have focused on orthologs of SoxE and β-catenin, key players of the male and female gonadic differentiation respectively in mammals. Based on phylogenetic analyses, Cg-SoxE, the oyster's SoxE ortholog, is closely related to vertebrate SoxE genes while Cg-β-catenin, the oyster's β-catenin ortholog, is classified amongst lophotrochozoa. The mRNA expression measured by qPCR in the gonadic area during a gametogenetic cycle is maximal for Cg-SoxE when sex is indiscernible and for Cg-β-catenin in mature females. Both expressions are localized from early germ cells to spermatocytes and pre-vitellogenic oocytes, and potentially in somatic cells. Cg-β-catenin is also expressed in vitellogenic oocytes. These actors may be involved in early oyster gonadic differentiation, which includes sex determination. Our results enhance the understanding of sex determination in C. gigas and in mollusks and they provide additional knowledge in compared genomics of reproduction and in molecular phylogeny. PMID:24120488
Ghude, Pranjali; Schallenberger, Mark A.; Fleming, Aaron M.; Muller, James G.; Burrows, Cynthia J.
As the most readily oxidized of DNA’s four natural bases, guanine is a prime target for attack by reactive oxygen species (ROS) and transition metal-mediated oxidants. The oxidation products of a modified guanosine nucleoside and of a single-stranded oligodeoxynucleotide, 5′-d(TTTTTTTGTTTTTTT)-3′ have been studied using oxidants that include CoII, NiII, and IrIV compounds as well as photochemically generated oxidants such as sulphate radical, electron-transfer agents (riboflavin) and singlet ...
Pedersen, G; Andresen, Lars; Matthiessen, M W; Rask-Madsen, J; Brynskov, J
cultures were stimulated with a synthetic CpG-oligodeoxynucleotide (ODN), exhibiting strong immunostimulatory effects in B cells. Interleukin (IL)-8 secretion was determined by enzyme-linked immunosorbent assay, nuclear factor-kappaB (NF-kB) activity by electrophoretic mobility shift assay and Ik......-8, apparently in the absence of NF-kB activation. Primary epithelial cells isolated from normal human colon expressed TLR9 mRNA, but were completely unresponsive to CpG-ODN stimulation in vitro. In conclusion, differentiated human colonic epithelial cells are unresponsive to TLR9 ligand stimulation...
Kumar, Sanjai; Jones, Trevor R.; Oakley, Miranda S.; Zheng, Hong; Shanmuga P Kuppusamy; Taye, Alem; Krieg, Arthur M.; Stowers, Anthony W; Kaslow, David C.; Hoffman, Stephen L.
Unmethylated CpG dinucleotide motifs present in bacterial genomes or synthetic oligodeoxynucleotides (ODNs) serve as strong immunostimulatory agents in mice, monkeys and humans. We determined the adjuvant effect of murine CpG ODN 1826 on the immunogenicity and protective efficacy of the Saccharomyces cerevisiae-expressed 19-kDa C-terminal region of merozoite surface protein 1 (yMSP119) of the murine malaria parasite Plasmodium yoelii. We found that in C57BL/6 mice, following sporozoite challe...
Weiner, George J; Liu, Hsin-Ming; Wooldridge, James E.; Dahle, Christopher E.; Krieg, Arthur M.
Recent advances in our understanding of the immune response are allowing for the logical design of new approaches to cancer immunization. One area of interest is the development of new immune adjuvants. Immunostimulatory oligodeoxynucleotides containing the CpG motif (CpG ODN) can induce production of a wide variety of cytokines and activate B cells, monocytes, dendritic cells, and NK cells. Using the 38C13 B cell lymphoma model, we assessed whether CpG ODN can function as immune adjuvants in...
To observe the liver condition of hyperthyroid patients with different course of disease, the levels of serum CG over night fasting and of 2 hours after meal in 132 hyperthyroid patients and 50 normal persons were measured by RIA. Hyperthyroid patients were divided into 3 groups by their different course of disease. For normal persons and the group whose course of disease within half year there was no significant difference in CG levels of fasting and 2 hours after meal. For the group whose course of disease in period of 0.5-2 years the CG level of 2 hours after meal were over the normal range (P<0.01), but no difference was found in CG level of fasting. For the group whose course of disease over 2 years, the CG levels of fasting and 2 hours after meal were all over the normal range (P<0.01, P<0.001), CG level of the 2 hours after meal was much higher. The results suggested that CG level measured 2 hours after meal was more sensitive than that after over night fasting in hyperthyroid patients with hepatocellular damage, and the damage degree paralleled to their courses of disease
Jurk, Marion; Vollmer, Jörg
Vertebrate toll-like receptors (TLRs) sense invading pathogens by recognizing bacterial and viral structures and, as a result, activate innate and adaptive immune responses. Ten human functional TLRs have been reported so far; three of these (TLR7, 8, and 9) are expressed in intracellular compartments and respond to single-stranded nucleic acids as natural ligands. The pathogen structure selectively recognized by TLR9 in bacterial or viral DNA was identified to be CpG dinucleotides in specific sequence contexts (CpG motifs). Short phosphorothioate-stabilized oligodeoxynucleotides (ODNs) containing such motifs are used as synthetic TLR9 agonists, and different classes of ODN TLR9 agonists have been identified with distinct immune modulatory profiles. The TLR9-mediated activation of the vertebrate immune system suggests using such TLR9 agonists as effective vaccine adjuvants for infectious disease, and for the treatment of cancer and asthma/allergy. Immune activation by CpG ODNs has been demonstrated to be beneficial in animal models as a vaccine adjuvant and for the treatment of a variety of viral, bacterial, and parasitic diseases. Antitumor activity of CpG ODNs has also been established in numerous mouse models. In clinical vaccine trials in healthy human volunteers or in immunocompromised HIV-infected patients, CpG ODNs strongly enhanced vaccination efficiency. Most encouraging results in the treatment of cancers have come from human phase I and II clinical trials using CpG ODNs as a tumor vaccine adjuvant, monotherapy, or in combination with chemotherapy. Therefore, CpG ODNs represent targeted immune modulatory drugs with a broad range of potential applications. PMID:18020622
Full Text Available Cationic liposomes are used for cellular delivery of antisense oligodeoxynucleotide (AsODN, where release of encapsulated AsODN is mainly controlled by endocytosis and fusion mechanisms. In this investigation, it was tried to model such a release process that is difficult to evaluate in cell culture. For this purpose, an AsODN model (against protein kinase C-α was encapsulated in a DODAP-containing cationic liposome and evaluated for size, zeta-potential, encapsulation and ODN stability. Vesicular models of outer layer and total plasma membranes and early and late endosomal membranes were developed, based on lipid content and pH, using ether injection method. ODN release was determined by the fluorescence dequenching of encapsulated FITC-ODN. Zeta potential, size and ODN encapsulation efficiency of the prepared liposomes were -2.49 ± 7.15 mV, 108.4 nm and 73% respectively. ODN protection was 3-4 times more than that of conventional liposome/ODN complexation method. There was a correlation between model concentration and percent of ODN release. At 7.5 µM, the percent of released ODN was 76% for the cholesterol-free model of the late endosome and 16% for the early endosomal membrane; while the release was less than 11% for the models of plasma membrane. ODN release increased with temperature in the range of 4-37◦C for the late endosomal model, but not for others, possibly due to their high cholesterol contents or acidic pH. The interaction was fast and completed within 5 minutes and didn’t change in the range of 5-60 minutes. Our data are in agreement with published cell culture studies and reveal that cell-liposomes interaction can be modeled by lamellar membranes.
Full Text Available the bone marrow and the intestine are the major sites of ionizing radiation (IR-induced injury. Our previous study demonstrated that CpG-oligodeoxynucleotide (ODN treatment mitigated IR-induced bone marrow injury, but its effect on the intestine is not known. In this study, we sought to determine if CpG-ODN have protective effect on IR-induced intestine injury, and if so, to determine the mechanism of its effect.Mice were treated with CpG-ODN after IR. The body weight and survival were daily monitored for 30 days consecutively after exposure. The number of surviving intestinal crypt was assessed by the microcolony survival assay. The number and the distribution of proliferating cell in crypt were evaluated by TUNEL assay and BrdU assay. The expression of Bcl-2, Bax and caspase-3 in crypt were analyzed by Immunohistochemistry assay. The findings showed that the treatment for irradiated mice with CpG-ODN diminished body weight loss, improved 30 days survival, enhanced intestinal crypts survival and maintained proliferating cell population and regeneration in crypt. The reason might involve that CpG-ODN up-regulated the expression of Bcl-2 protein and down-regulated the expression of Bax protein and caspase-3 protein.CpG-ODN was effective in protection of IR-induced intestine injury by enhancing intestinal crypts survival and maintaining proliferating cell population and regeneration in crypt. The mechanism might be that CpG-ODN inhibits proliferating cell apoptosis through regulating the expression of apoptosis-related protein, such as Bax, Bcl-2 and caspase-3.
Chitosan–silica/CpG oligodeoxynucleotide (ODN) nanohybrids were synthesized to stimulate Toll-like receptor 9-mediated induction of interleukin-6 (IL-6). The chitosan–silica hybrid was first synthesized from a mixture of chitosan and 3-glycidoxypropyl trimethoxysilane under acidic conditions via a sol–gel process, and then used to condense CpG ODN2006x3-PD to yield chitosan–silica/CpG ODN nanohybrids. Scanning electron microscopy and atomic force microscopy showed that the chitosan–silica/CpG ODN nanohybrids had an elliptic shape with a diameter of 100–200 nm. After soaking in HAc–NaAc buffer solution (pH 5.5), the nanohybrids exhibited sustained release of CpG ODN. When the nanohybrids were separately exposed to 293XL-hTLR9 cells and peripheral blood mononuclear cells, no significant toxicity was observed. An immunochemical assay for cellular uptake revealed that the nanohybrids were taken up by the cells and located in endolysosomes. An enzyme-linked immunosorbent assay for cytokines indicated that the nanohybrids effectively stimulated the induction of IL-6. Chitosan–silica/CpG ODN nanohybrids underwent cellular uptake and enhanced induction of IL-6 to a greater degree than conventional chitosan/CpG ODN nanocomplexes, indicating that they have an enhanced delivery efficiency. - Highlights: • Chitosan-silica/CpG ODN nanohybrids (100 nm–200 nm) were synthesized. • Nanohybrids exhibited a sustained release behavior for CpG ODN. • Cellular uptake of nanohybrids was enhanced due to presence of silica. • The released CpG ODN stimulated production of cell cytokine
Ziegler, Saskia; Eberle, Mariel Esther; Wölfle, Sabine J; Heeg, Klaus; Bekeredjian-Ding, Isabelle
MicroRNAs (miRNAs) are fine-tuners in cellular processes, including those of the immune response. To study their functions and effects in immune cells, it is necessary to achieve specific silencing of individual miRNAs. To date, introduction of antisense microRNAs (antagomiRs) into primary cells is based on electroporation, lipofection, and viral vectors. However, these techniques often compromise viability, proliferative capacity, and differentiation. Furthermore, efficiency varies depending on the cell type and some are not suitable for in vivo approaches. To overcome these limitations we exploited the property of phosphorothioate (PTO)-modified DNA oligodeoxynucleotides (ODN) to enter cells with high efficacy: we developed and evaluated ODN/antagomiR constructs that consist of a PTO-ODN carrier covalently linked to a fully methylated antagomiR RNA sequence. Using these constructs, we achieved transfection efficiency of approximately 99% in leukocytes-in particular, in B lymphocytes that are hard to transfect with other methods. Our data demonstrate that miRNA silencing by the antagomiR portion of the constructs was specific and efficient, which could be further confirmed by an increase in target protein under silencing conditions. The constructs were successfully tested in human B cells, plasmacytoid dendritic cells, monocytes, and monocyte-derived dendritic cells, thus demonstrating their versatility. Moreover, introduction of stimulatory CpG sequences into the ODN portion conveys immune stimulatory quality when intended. Thus, bifunctional ODN/antagomiR constructs represent a highly efficient, versatile, and easy-to-handle tool to manipulate cellular miRNA expression levels and to allow the subsequent investigation of specific miRNA functions. PMID:24236889
Zhuang, X B; Xing, N; Zhang, Q; Yuan, S J; Chen, W; Qiao, T K
Immunotherapy is a hot issue in cancer research over the years and tumor cell vaccine is one of the increasing number of studies. Although the whole tumor cell vaccine can provide the best source of immunizing antigens, there is still a limitation that most tumors are not naturally immunogenic. CpG Oligodeoxynucleotides (CpG ODNs), synthetic oligonucleotides containing a cytosine-phosphate-guanine(CpG) motif, was shown to enhance immune responses to a wide variety of antigens. In this study, we generated the radioresistant Lewis lung cancer cell by repeated X-ray radiation and inactivated it as a whole tumor cell vaccine to enhance the immunogenicity of tumor cell vaccine. Mice were subcutaneously immunized with this inactivated vaccine combined with CpG ODN1826 and then inoculated with autologous Lewis lung cancer (LLC) to estimate the antitumor efficacy. The results showed that the radioresistant tumor cell vaccine combined with CpG ODN1826 could significantly inhibit tumor growth, increased survival of the mice and with 20% of the mice surviving tumor free in vivo compared with the unimmunized mice bearing LLC tumor. A significant increase of apoptosis was also observed in the tumor prophylactically immunized with vaccine of inactivated radioresistant tumor cell plus CpG ODN1826. The potent antitumor effect correlated with higher secretion levels of tumor necrosis factor-alpha(TNF-α) and lower levels of interleukin-10(IL-10) concentration in serum. Furthermore, the results suggested that the antitumor mechanism was probably depended on the decreased level of programmed death ligand-1(PD-L1) which plays an important role in the negative regulation of immune response by the inhibition of tumor antigen-specific T cell activation. These findings clearly demonstrated that the radioresistant tumor cell vaccine combined with CpG ODN1826 as an appropriate adjuvant could induce effective antitumor immunity in vivo. PMID:26458317
Cheng, Wing Ki; Plumb, Adam William; Lai, Jacqueline Cheuk-Yan; Abraham, Ninan; Dutz, Jan Peter
Current influenza vaccines generate humoral immunity, targeting highly variable epitopes and thus fail to achieve long-term protection. T cells recognize and respond to several highly conserved epitopes across influenza serotypes. A strategy of raising strong cytotoxic T cell memory responses to epitopes conserved across serotypes would provide cross serotype protection, eliminating the need for annual vaccination. We explored the adjuvant potential of epicutaneous (ec) and subcutaneous (sc) delivery of CpG oligodeoxynucleotide in conjunction with sc protein immunization to improve protection against influenza A virus (IAV) infections using a mouse model. We found enhanced long-term protection with epicutaneous CpG ODN (ecCpG) compared to subcutaneous CpG ODN (scCpG) as demonstrated by reduced viral titers in the lungs. This correlated with increased antigen-specific CD8 T cells in the airways and the lungs. The memory T cell response after immunization with ecCpG adjuvant was comparable to memory response by priming with IAV infection in the lungs. In addition, ecCpG was more efficient than scCpG in inducing the generation of IFN-γ producing CD4 T cells. The adjuvant effect of ecCpG was accompanied with its ability to modulate tissue-homing molecules on T cells that may direct them to the site of infection. Together, this work provides evidence for using ecCpG to induce strong antibody and memory T cell responses to confer protection against IAV infection. PMID:27524984
Mohammadi, Milad; Aamodt, Tor M.; Dally, William J.
We introduce the Coarse-Grain Out-of-Order (CG- OoO) general purpose processor designed to achieve close to In-Order processor energy while maintaining Out-of-Order (OoO) performance. CG-OoO is an energy-performance proportional general purpose architecture that scales according to the program load. Block-level code processing is at the heart of the this architecture; CG-OoO speculates, fetches, schedules, and commits code at block-level granularity. It eliminates unnecessary accesses to ener...
Yamanaka, T; Komatani, H; Meshi, T; Naito, S; Ishikawa, M; Ohno, T
Tobacco mosaic virus (TMV)-Cg is a crucifer-infecting tobamovirus that was isolated from field-grown garlic. We determined the complete nucleotide sequence of the genomic RNA of TMV-Cg. The genomic RNA of TMV-Cg consists of 6303 nucleotides and encodes four large open reading frames, organized basically in the same way as that of other tobamoviruses. The nucleotide and deduced amino acid sequences are very similar to those of the other crucifer-infecting tobamoviruses that have been sequenced so far. PMID:9608662
18F fluorodeoxyglucose with PET(18FDG-PET) imaging is used to image tumors. Due to the high cost and low availability, PET is not accessible to many patients. Labelled 99mTc agents to be used with SPECT could be an alternative to 18FDG-PET. Glucosamine (G), a six-carbon amino sugar, enters the cells by a glucose transporter system and by phosphate and glycolytic pathways forms glucosamine-6-phosphate. Reacting G with cysteine (C) labeled with 99mTc (99mTcCG) could be used to show with SPECT hypermetabolic lesions similar to 18FDG-PET imaging. The aim of this study was to evaluate the imaging with 99mTc-CG in detecting tumors and correlate them with pathology. Material and Methods: Labelling of 99mTc-CG was done by adding (C), (G), N-hydroxysuccinimide, carbodiimide, and stannous chloride to the 99mTc. Patients: 35 patients (23 F, 12 M, range age 7-79 years, mean 52 ) with histopathology diagnosis were referred for scintigraphy with 99mTc-CG. Imaging: Patients fasted for 12 hr, and 1-4 hours after the i.v. injection of 920-1088 MBq of 99mTc-CG, underwent 1) whole body scan, 2) SPECT images (reconstructed with OSEM algorithm) on the field of view of the lesion. To eliminate the intravascular and interstitial activity of the 99mTcC-G imaging, patients were given 737 MBq of 99mTc human serum albumin(99mTc-HSA) and other SPECT was done. We normalized both SPECT images and subsequently the 99mTc-HSA image was subtracted from the 99mTc-CG image. All images were visually analysed. Results: 32 lesions were malignant (breast (5), colon (8), lung (9), Hodgkin's disease(6), thyroid(1), bladder (1), melanoma (1), ependimoma (1), and 3 were benign (brain radiation necrosis, breast inflammations and inespecific granuloma). 99mTc C-G show abnormal focal uptake in all malignant lesions (sensitivity=100%) as well as in two benign lesion (false positive). Accuracy of 99mTcC-G in patients with malign tumors was 94%. SPECT images defined the tumor uptake better than planar imaging
Ying-An Jiang; He-Sheng Luo; You-Yuan Zhang; Li-Fang Fan; Chong-Qing Jiang; Wei-Jin Chen
AIM: To evaluate the effect of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynudeotide (As-ODN) on telomerase activity and cell apoptosis in colon cancer cell line SW480.METHODS: As-ODN was transfected into cells SW480 by liposomal transfection. Cultured cells were divided into three groups: ASODN (5′GGAGCGCGCGGCATCGCGGG-3), sense oligodeoxynucleotide (5′-CCCGCGATGCCGCGCGCTCC-3; SODN) and control. The concentration of oligodeoxynucleotide and lipsome was 10 μmol/L and 16 mg/L, respectively. The activity of telomerase was examined by telomeric repeat amplification protocol (TRAP)-enzyme-linked immunosorbent assay (ELISA), and cell apoptosis was observed by morphology and flow cytometry in each group.RESULTS: Telomerase activity began to be down-regulated or inhibited when cells SW480 were treated with As-ODN for 72 h, and cell apoptosis was induced.CONCLUSION: It is suggested that hTERT As-ODN might specially inhibit the activity of telomerase in colon cancer cells and it is further proved that the hTERT gene has a significant correlation with telomerase activity. Further evidence is needed to prove whether hTERT As-ODN is a potential tool for the treatment of colon cancer.
Xiaohuai XUE; Luhai WU; Bainian QIAN; Jingli LI; Songnian LOU
X80 pipeline steel produced by TMCP has high strength and high toughness with ultrafine grain microstructure. The microstructure coarsens and the toughness worsens at the coarse grained (CG) HAZ apparently after weld simulation. The experimental results indicated that the bainitic ferrite and the second phases formed at cooling are differently as the variation of carbon in base metal. In low carbon steels, the bainitic ferrite laths are long and narrow, the second phases are complex including residual austenite, martensite, the M-A constituent and the Fe3C carbide. The formation of Fe3C carbide is the main reason of the poor toughness in CG HAZ. The ultralow carbon in base metal, however, can improve the CG HAZ toughness through restraining the formation of carbides, decreasing the M-A constituent, increasing the residual austenite content, which are beneficial to the CG HAZ toughness.
Tölzer, Christine; Pal, Sonia; Watzlawick, Hildegard; Altenbuchner, Josef; Niefind, Karsten
The enzyme cgHle from C. glutamicum, which has acetyl ester hydrolase activity, was crystallized in four different crystal forms. X-ray diffraction data have been collected to a resolution of 1.2 Å.
Serial βhCG testing can be a helpful tool in deciding how to manage pregnancy of unknown location. Its use in emergency gynaecology clinics can prevent unnecessary admission and intervention. However, despite NICE Guidelines on when it is safe to opt for conservative management, it was identified that there was a problem with over-testing of βhCG when patients could be discharged with instructions to repeat a urinary pregnancy test in two weeks. Two PDSA cycles were undertaken to improve the awareness of NICE guidelines: the first involved formal and informal educational sessions and the second involved the inclusion of a guideline summary on the front of patients' notes when they were having serial βhCG tests for doctors to refer to. Case notes were reviewed for 157 women who had βhCG tests at baseline and 48 hours. Of these, 139 were suitable for serial βhCG testing, and 83 of these were suitable for discharge after 48 hours. Of the 83 patients that were eligible for discharge, there were 31 unnecessary βhCG tests done, 23 of which were prior to intervention. A significant improvement was noted, with between 4-10 unnecessary βhCG tests per fortnight prior to intervention, 0-3 following the first intervention, and 0-2 following the second. Reduction in unnecessary βhCG testing has positive implications for patients, who do not have to take unnecessary time off work, prolong an already very distressing period, and have unnecessary blood tests. There are also cost and time saving implications for the hospital. PMID:26893889
In this paper, we calculate the top quark rare decays t → cγ and t → cg in an extension of the standard model, where baryon number and lepton number are local gauge symmetries. Adopting reasonable assumptions on the parameter space, we find that the branching ratios of t → cγ and t → cg can reach 10−6 and 10−5 respectively, which can be detected in near future. (physics of elementary particles and fields)
Zahid Naseer; Ejaz Ahmad; Nemat Ullah; Muhammad Yaqoob; Zeeshan Akbar
Objective: To determine the effect of equine chorionic gonadotropin (eCG) and controlled internal drug releasing (CIDR) on estrus response, ovulation and pregnancy rate in Nili-Ravi buffaloes under field condition. Methods: Twenty anestrous buffaloes with more than 150 days postpartum were used in this study. To confirm anestrous condition, ovarian status and serum progesterone concentration were determined before the start of study. Buffaloes were randomly divided into eCG (n=10) and CIDR (n=10) groups. eCG group were treated with eCG (Chronogest 1000 I.U. i.m.), while CIDR group received CIDR devices for 7 days. All buffaloes in CIDR group were injected PGF2α (0.25 ug/mL; 2 mL; i.m.) one day before CIDR removal. Buffaloes were observed for estrus visually and were inseminated with frozen-thawed semen at detected estrus. Ovulation was confirmed by presence of CL 12 day post AI. Pregnancy diagnosis was done 50 days post AI. Results: Estrus response, ovulation rate and pregnancy rate were similar (P>0.05) among eCG and CIDR groups (90% vs. 80%; 78% vs. 75% and 50% vs. 60% respectively). Conclusion: It is concluded that both CIDR and eCG protocols are effective and promising remedies for the anestrous buffaloes under commercial as well as under field conditions.
The enzyme cgHle from C. glutamicum, which has acetyl ester hydrolase activity, was crystallized in four different crystal forms. X-ray diffraction data have been collected to a resolution of 1.2 Å. CgHle is an enzyme that is encoded by gene cg0961 from Corynebacterium glutamicum. The physiological function of cgHle is so far unclear. Bioinformatic annotations based on sequence homology indicated that cgHle may be an acetyl-CoA:homoserine acetyl transferase and as such may be involved in methionine biosynthesis, but recent evidence has shown that it is an esterase that catalyzes the hydrolysis of acetyl esters. Here, the crystallization of cgHle in two orthorhombic crystal forms, a trigonal crystal form and a monoclinic crystal form is described. The trigonal crystals have a solvent content of 83.7%, which is one of the highest solvent contents ever found for protein crystals. One of the orthorhombic crystals diffracted X-rays to at least 1.2 Å resolution
CG(Computer Graphics)作为商品,带有一定的世俗性或曰通俗性。通过商业传播,CG得以在世界各地快速地普及。但是,单纯地从大众传播学角度来看待CG作品,其审美意义未免有些片面。本文对CG的发展、CG的制作流程、色彩心理的接受程度和艺术品的商业生产及其发展前景等进行叙述,让读者对夹杂在商业和纯艺术之间的CG艺术有一个更明确的认识。%As a commodity, CG has certain secularism or popularity. CG was able to quickly spread around the world, which traveled through business. However, purely from the perspective of mass communication to analyze CG work, its aesthetic significance is one-si-ded. Through the description of the development of CG, the essence of its production process,color psychological acceptance, and com-mercial production of art and its developmental prospects etc. , the paper would let the reader have a clear understanding about CG be-tween commodity and pure art.
Li, Meijia; Wang, Lingling; Qiu, Limei; Wang, Weilin; Xin, Lusheng; Xu, Jiachao; Wang, Hao; Song, Linsheng
Glutamic acid decarboxylase (GAD), a rate-limiting enzyme to catalyze the reaction converting the excitatory neurotransmitter glutamate to inhibitory neurotransmitter γ-aminobutyric acid (GABA), not only functions in nervous system, but also plays important roles in immunomodulation in vertebrates. However, GAD has rarely been reported in invertebrates, and never in molluscs. In the present study, one GAD homologue (designed as CgGAD) was identified from Pacific oyster Crassostrea gigas. The full length cDNA of CgGAD was 1689 bp encoding a polypeptide of 562 amino acids containing a conserved pyridoxal-dependent decarboxylase domain. CgGAD mRNA and protein could be detected in ganglion and hemocytes of oysters, and their abundance in hemocytes was unexpectedly much higher than those in ganglion. More importantly, CgGAD was mostly located in those granulocytes without phagocytic capacity in oysters, and could dynamically respond to LPS stimulation. Further, after being transfected into HEK293 cells, CgGAD could promote the production of GABA. Collectively, these findings suggested that CgGAD, as a GABA synthase and molecular marker of GABAergic system, was mainly distributed in hemocytes and ganglion and involved in neuroendocrine-immune regulation network in oysters, which also provided a novel insight to the co-evolution between nervous system and immune system. PMID:27208883
An oligonucleotide that can form a triplex with double-stranded DNA is called a triplex-forming oligonucleotide (TFO). TFOs have gained considerable attention because of their potential as gene targeting tools. However, triplex DNA formation involves inherent problems for practical use. The most important problem is that natural nucleotides in TFO do not have sufficient affinity and base pair-selectivity to pyrimidine-purine base pair, like a CG or TA base pair, within dsDNA. This suggests that dsDNA region including a CG or TA base pair cannot be targeted. Therefore, artificial nucleotides, especially with non-natural nucleobases, capable of direct recognition of a CG or TA base pair via hydrogen bond formation have been developed; however, nucleotides with better selectivity and stronger affinity are necessary for implementing this dsDNA-targeting technology using TFOs. Under such a background, we considered that facile and efficient synthesis of various nucleobase derivatives in TFOs would be useful for finding an ideal nucleobase for recognition of a CG or TA base pair because detailed and rational exploration of nucleobase structures is facilitated. Recently, to develop a nucleobase recognizing a CG base pair, we have used post-elongation modification, i.e., modification after oligonucleotide synthesis, for the facile synthesis of nucleobase derivatives. This review mainly summarizes our recent findings on the development of artificial nucleobases and nucleotides for recognition of a CG base pair in triplexes formed between dsDNA and TFOs. PMID:24189561
王正全; 李金玲; 纪道松; 李涛
东方红CG702果园型履带拖拉机适应于我国广大农村小面积农田、山坡地、果园以及农田的耕作，以果园工作为主提高其综合利用率。东方红CG702是一种适合国内外果园、坡地作业需求的全新半刚性履带拖拉机，多数性能指标等同或接近国外先进水平。%The Dongfanghong CG702 crawler tractor of orchard type is suit for small field、hillside, orchards farming, It is mainly working for orchard work to improve the comprehensive utilization rate. The Dongfanghong CG702 crawler tractor of orchard type is suitable for domestic and overseas orchards, sloping fields operational requirements and it is the new semi-rigid crawler tractors, most performance equal to or close to the international advanced level.
Hai-yan DU; Zhong-ming TANG; Hai-feng SONG; Li-hou DONG; Bi-jun ZHAO; Jie FU; Qing-qing WANG; FangCHEN; Lun OU; Na LI; Xiao SUN
Aim:DNAs containing unmethylated CpG motifs can stimulate innate and adaptive immunity.The aim of this study was to investigate the immunostimulatory and anti-neoplasm effects of a novel CpG oligodeoxynucleotide,ODN10,in tumor-bearing mice.Methods:B16 melanoma-bearing C57BL/6 mice were administered ip or sc with ODN10 or conventional CpG ODN1826 on the indicated days post inoculation.The animal survival rate and the inhibitory effect on tumor growth were observed in vivo.B and T lymphocyte proliferation,natural killing cell cytotoxicity and the phagocytic ability of peritoneal macrophages from the animals were determined using [3H]-thymidine incorporation assay,4-h 51Cr release assay and neutral red chromometry method,respectively.The serum levels of IL-12,IL-4,and IgE were quantified using ELISA assays.Histological examination of tumor tissues was performed after HE staining,and the expression of PCNA,CD63,and CD80 in tumor tissues was analyzed with immunohistochemistry.Results:ODN10 (1,5,and 25 mg/kg) significantly inhibited the growth and metastasis of the tumor,and significantly prolonged the survival of tumor-bearing mice,as compared with ODN1826,The immune status was suppressed in tumor-bearing mice.Both ODN10 and ODN1826 significantly reversed the suppressed immunoactivities in tumor-bearing mice,which included promoting B and T lymphocyte proliferation,enhancing NK cell and peritoneal macrophage activities,inducing IL-12 secretion and inhibiting IL-4 and IgE secretion.Further,CpG ODNs decreased PCNA and CD63 expression while induced expression of CD80.ODN10 presented more potent activity,and displayed the most prominent immunostimulatory potential.Conclusion:ODN10 produces prominent immunomodulatory effects on cellular immunity in tumor-bearing mice,which might help reverse the established Th2-type responses to the Th1-type responses,thus may be used as a potent anti-tumor immunotherapy agent or adjuvant.
Law Chung HIONG; Kiew Lik VOON; Nor Azizan ABDULLAH; Munavvar A SATTAR; Nazarina AbduRAHMAN; Abdul Hye KHAN; Edward James JOHNS
Aim:The aim of the present study was to investigate the effectiveness of trans-forming growth factor (TGF)-β1 antisense oligodeoxynucleotides (ODN) in ame-liorating deteriorated kidney function in rats with puromycin-induced chronic renal failure (CRF). Methods:Saline, puromycin, puromycin+TGF-β1 antisense ODN or puromycin+scrambled ODN were administered to unilaterally nephrecto-mized rats. Renal hemodynamic and excretory measurements were taken in the anaesthetized rats that had undergone surgical procedure. Results:It was ob-served that in the CRF rats, there was a marked reduction in the renal blood flow (RBF), glomerular filtration rate (GFR), severe proteinuria, and almost 6-fold in-creased fractional excretion of sodium (FE Na+) as compared to that in the control rats (all P<0.05). It was further observed that in the CRF rats, the treatment with TGF-β1 antisense, but not scrambled ODN, markedly attenuated the reduction of RBF, GFR, and proteinuria and markedly prevented the increase of the FE Na+ (all P<0.05). In addition, the renal hypertrophy in the CRF group (P<0.05 vs non-renal failure control) was markedly attenuated after treatment with TGF-1 antisense ODN (P<0.05). Focal segmental glomerulosclerosis was evident only in the un-treated and scrambled ODN-treated CRF groups. An interesting observation of this study was that in the CRF rats, although there was marked attenuating and preventive effects of the TGF-β1 antisense ODN on the deteriorated renal functions, the antisense treatment did not cause any marked change in the renal expression of TGF-β1 at the protein level. Conclusion:Collectively, the data obtained sug-gests that TGF-β1 antisense ODN possesses beneficial effects in puromycin-induced chronic renal failure and that the deterioration in morphology and im-paired renal function in this pathological state is in part dependent upon the action of TGF-β1 within the kidney.
Full Text Available Nobutaka HanagataNanotechnology Innovation Station, National Institute for Materials Science, Tsukuba, Ibaraki, and Graduate School of Life Science, Hokkaido University, Kita-ku, Sapporo, JapanAbstract: Unmethylated cytosine-phosphate-guanosine (CpG oligodeoxynucleotides (ODNs are recognized by Toll-like receptor 9 (TLR9 found in antigen-presenting cells and B cells and can activate the immune system. Using CpG ODNs as an adjuvant has been found to be effective for treating infectious diseases, cancers, and allergies. Because natural ODNs with only a phosphodiester backbone are easily degraded by nuclease (deoxyribonuclease [DNase] in serum, CpG ODNs with a phosphorothioate backbone have been studied for clinical application. CpG ODNs with a phosphorothioate backbone have raised concern regarding undesirable side effects; however, several CpG ODNs with only a phosphodiester backbone have been reported to be stable in serum and to show an immunostimulatory effect. In recent years, research has been conducted on delivery systems for CpG ODNs using nanoparticles (NPs. The advantages of NP-based delivery of CpG ODN include (1 it can protect CpG ODN from DNase, (2 it can retain CpG ODN inside the body for a long period of time, (3 it can improve the cellular uptake efficiency of CpG ODN, and (4 it can deliver CpG ODN to the target tissues. Because the target cells of CpG ODN are cells of the immune system and TLR9, the receptor of CpG ODN is localized in endolysosomes, CpG ODN delivery systems are required to have qualities different from other nucleic acid drugs such as antisense DNA and small interfering RNA. Studies until now have reported various NPs as carriers for CpG ODN delivery. This review presents DNase-resistant CpG ODNs with various structures and their immunostimulatory effects and also focuses on delivery systems of CpG ODNs that utilize NPs. Because CpG ODNs interact with TLR9 and activate both the innate and the adaptive immune
Song, Xiaorui; Wang, Hao; Chen, Hao; Sun, Mingzhe; Liang, Zhongxiu; Wang, Lingling; Song, Linsheng
Hemocytes are the effective immunocytes in bivalves, which have been reported to be derived from stem-like cells in gill epithelium of oyster. In the present work, a conserved haematopoietic transcription factor Tal-1/Scl (Stem Cell Leukemia) was identified in Pacific oyster (Cg-SCL), and it was evolutionarily close to the orthologs in deuterostomes. Cg-SCL was highly distributed in the hemocytes as well as gill and mantle. The hemocyte specific genes Integrin, EcSOD and haematopoietic transcription factors GATA3, C-Myb, c-kit, were down-regulated when Cg-SCL was interfered by dsRNA. During the larval developmental stages, the mRNA transcripts of Cg-SCL gradually increased after fertilization and peaked at early trochophore larvae stage (10 hpf, hours post fertilization), then sharply decreased in late trochophore larvae stage (15 hpf) before resuming in umbo larvae (120 hpf). Whole-mount immunofluorescence assay further revealed that the immunoreactivity of Cg-SCL appeared in blastula larvae with two approximate symmetric spots, and this expression pattern lasted in gastrula larvae. By trochophore, the immunoreactivity formed a ring around the dorsal region and then separated into two remarkable spots at the dorsal side in D-veliger larvae. After bacterial challenge, the mRNA expression levels of Cg-SCL were significantly up-regulated in the D-veliger and umbo larvae, indicating the available hematopoietic regulation in oyster larvae. These results demonstrated that Cg-SCL could be used as haematopoietic specific marker to trace potential developmental events of hematopoiesis during ontogenesis of oyster, which occurred early in blastula stage and maintained until D-veliger larvae. PMID:26915307
Trifonov Edward N
Full Text Available Abstract Background The periodical occurrence of dinucleotides with a period of 10.4 bases now is undeniably a hallmark of nucleosome positioning. Whereas many eukaryotic genomes contain visible and even strong signals for periodic distribution of dinucleotides, the human genome is rather featureless in this respect. The exact sequence features in the human genome that govern the nucleosome positioning remain largely unknown. Results When analyzing the human genome sequence with the positional autocorrelation method, we found that only the dinucleotide CG shows the 10.4 base periodicity, which is indicative of the presence of nucleosomes. There is a high occurrence of CG dinucleotides that are either 31 (10.4 × 3 or 62 (10.4 × 6 base pairs apart from one another - a sequence bias known to be characteristic of Alu-sequences. In a similar analysis with repetitive sequences removed, peaks of repeating CG motifs can be seen at positions 10, 21 and 31, the nearest integers of multiples of 10.4. Conclusions Although the CG dinucleotides are dominant, other elements of the standard nucleosome positioning pattern are present in the human genome as well. The positional autocorrelation analysis of the human genome demonstrates that the CG dinucleotide is, indeed, one visible element of the human nucleosome positioning pattern, which appears both in Alu sequences and in sequences without repeats. The dominant role that CG dinucleotides play in organizing human chromatin is to indicate the involvement of human nucleosomes in tuning the regulation of gene expression and chromatin structure, which is very likely due to cytosine-methylation/-demethylation in CG dinucleotides contained in the human nucleosomes. This is further confirmed by the positions of CG-periodical nucleosomes on Alu sequences. Alu repeats appear as monomers, dimers and trimers, harboring two to six nucleosomes in a run. Considering the exceptional role CG dinucleotides play in the
Rison, W.; Krehbiel, P. R.; Stock, M.; Godman, H.; Lu, G.; Cummer, S. A.
Upward-initiated cloud-to-ground (CG) lightning flashes are often triggered by preceding natural IC or CG flashes. Warner (2011) has recently documented (by means of high-speed video) upward-initiated negative CG flashes from tall towers triggered by preceding positive CG flashes. In this paper we discuss VHF lightning mapping images and ULF magnetic measurements of an upward-initiated positive CG flash triggered by a preceding negative CG flash. The VHF lightning mapping data are from a small (10 km diameter) seven-station Lightning Mapping Array (LMA) installed in northeastern Virginia. The ULF magnetic measurements were made at Duke University, 400 km away. The LMA data show a four-stroke negative CG flash is followed immediately by a negative leader which propagates from the ground upward through the lower positive thunderstorm charge (~4 km altitude) and the midlevel negative charge (~6 km altitude) into the upper positive charge (~8 km altitude). This is followed by about 500 ms of horizontal development of negative leaders through the upper positive charge, to the north and west of the upward leader. ULF magnetic data show positive charge moving downward during this time interval. (We cannot tell if the current is between the upper positive charge and the midlevel negative charge, or if the current flows through the negative leader channel to ground.) After this current cuts off, a positive CG stroke (with a 50 ms long continuing current) developed along a different channel, with an NLDN-detected ground strike point about 30 km NNW of the upward leader. The negative CG preceding this flash removed negative charge from the thunderstorm, "uncovering" the upper positive charge, and producing a rapid change in the electric field at the ground. The upward negative leader developed in response to this rapid field change. There are several tall objects near (within 1 km) the first LMA source detected from the upward leader. We cannot determine which of these
Ying-An Jiang; He-Sheng Luo; Li-Fang Fan; Chong-Qing Jiang; Wei-Jin Chen
AIM: To explore the effect of antisense oligodeoxynucleotide (As-ODN) of telomerase RNA on telomerase activity and cell apoptosis in human colon cancer.METHODS: As-ODN waS transfected into SW480 cells by liposomal transfection reagent. Telomerase activity of SW480 cells was examined by telomeric repeat amplification protocol (TRAP) and enzyme-linked immunosorbent assay (ELISA).Apoptosis was analyzed by morphology and flow cytometry.RESULTS: The telomerase activity in SW480 cells trandected with 1.0 μmol/L of As-ODN for 2-5 days, was significantly decreased in a time-dependent manner, and the cells underwent apoptosis. The missense ODN (Ms-ODN) and the control group transfected with SW480 cells did not show these changes.CONCLUSION: As-ODN can specifically inhibit the telomerase activity of SW480 cells and induce apoptosis.
Object To explore tile effect of lipofectin - c - erbB2 antisense oligodeoxynucleotides on radiosensitivity of human ovarian cancer cell llne. Methods The expression of c - erbB2 was detected by means of RT - PCR, cellular response to irradiation was evaluated by tile colony forming assay. Results Lipofectin- c - erbB2 antisense oligodeoxynucleotides（AS- ODN) could suppress the expression of c - erbB2 , and significantly decreased the colony forming rate of human ovarian cancer cells after ionizing irradiation （P 0.05 ). Condusion c - erbB2 antisense oligodeoxynueleotides sensitized the SKOV3 to ionizing irradiation through decreasing the expression of e - erbB2 , which might be the result of the fact that c - erbB2 antisense oligodeoxynueleotides inhibit the eelluar signal transductionpathway relating to the radiation- resistant phenotype.
A double stranded oligodeoxynucleotide containing a single 125I-dC in a defined location was used to investigate DNA strand breakage resulting from 125I decay. Samples of a 41 bp oligodeoxynucleotide were incubated in 20 mM phosphate buffer (PB), or PB plus 2 M dimethylsulphoxide (DMSO), at 4 C during 18-20 days. The 32P-5'-end labelled DNA fragments produced by 125I decays were separated on denaturing polyacrylamide gels, and the 32P activity in each fragment was determined by scintillation counting after elution of fragments from gel. Most of the breaks, around 90%, occurred within 4-5 nucleotides of the 125I-dC, but DNA breaks were detected up to 16 nucleotides from the decay site. The 125I-dC was located at the 21st nucleotide from the 32P-5'-end label, and since 32P was not detected in fragments longer than 20 nucleotides, it was assumed that all 125I decay events produce at least one break in the 125I-labelled DNA strand. The results show a considerable protection effect of DMSO on DNA breaks at sites >5-6 nucleotides from the 125I location. The probability of breaks in this region was decreased with DMSO by a factor of 2 to 8-fold, suggesting significant role for radical-mediated DNA breaks at the more distant sites. However, the total protection effect of DMSO is rather small: 1.1, because of the small contribution of breakage at distant sites to the total yield. (orig.)
Bannai, M; Ichikawa, M; Nishimura, F; Nishihara, M; Takahashi, M
One of the problems of introducing antisense oligodeoxynucleotides (ODN) into the central nervous system (CNS) is their rapid disappearance from the target site due to their dispersion and diffusion, which results in poor uptake and/or retention in cells (M. Morris, A.B. Lucion, Antisense oligonucleotides in the study of neuroendocrine systems, J. Neuroendocrinol. 7 (1995) 493-500; S. Ogawa, H.E. Brown, H.J. Okano, D.W. Pfaff, Cellular uptake of intracerebrally administrated oligodeoxynucleotides in mouse brain, Regul. Pept. 59 (1995) 143-149) [2,5]. Recently, we adapted a new method using water-absorbent polymer (WAP; internally cross-linked starch-grafted-polyacrylates) as a carrier for antisense ODN. The polymer forms a hydro-gel after absorbing water which is chemically and biologically inert. In these studies, the polymer (powder-form) is fully swollen by physiological saline containing antisense ODN (0.2 micromol/ml) to make 80-fold volume gel. Hydro-gel (1 microliter) is injected into the target site, and water solutes are assumed to be diffused stoichiometrically into CNS from the surface of the gel. Histological studies indicate that 24 h after the injection, antisense ODN (5'biotinylated-S-oligos of 15 mer) are distributed to within 800 micrometer from the edge of the area where the gel is located and then gradually disappear from this area within days, but still remain within 300-micrometer distance 7 days later. Antisense ODN are effectively incorporated by all the cell types examined, i.e., neurons, astrocytes and microglias, and suppress the synthesis of the target protein. This method can be adapted to slow delivery of antisense ODN and other water soluble substances into the CNS. PMID:9767125
Gabalda, G.; Bonvalot, S.; Hipkin, R.
A newly developed interactive computer program, CG3TOOL, has been dedicated to the processing of the gravity data acquired by the Scintrex CG-3/3M automated gravity meter. The aim of CG3TOOL is two fold: to allow for an objective evaluation of Scintrex data and to provide a higher resolution in data reductions than those computed in real time by the microprocessor-controlled instrument. The program reads the gravity data acquired in either field or cycle mode (field surveys and continuous recordings, respectively) and then downloaded from the meter to a PC computer. The processing tasks are divided into two successive levels. Level 1 is dedicated to the reduction of the daily data files by applying standard or accurate corrections (earth tide, instrumental drift, atmospheric pressure). The precise corrections are performed up to the microGal (μGal) level, in accordance with the specifications of high-resolution surveys. Level 2 contains a series of processing tools (including network adjustment, anomaly computation, and gravity meter calibration) that will precisely compute and adjust the gravity values with error estimates. The interactive procedures and the program output (plot and text files) have been designed to ease data handling and archiving as well as to provide useful information for future purposes of data interpretation or modeling. CG3TOOL was developed in a standard C language for Unix Sun workstations and uses the standard graphical and mathematical Generic Mapping Tools (GMT) free library, available from the web. The objectives and principles of the computer program are presented below along with corresponding examples of the main processing tasks applied to observed data.
Objective: To study the clinical significance of changes of serum levels of TBA, CG, HA in neonate with parenteral nutrition. Methods: Serum total bile acid (TBA, with biochemistry) and CG, HA (with RIA) contents were measured in 52 neonates (full-term 32, preterm 20) with parenteral nutrition and 28 neonates (full-term 16, preterm 12) without parenteral nutrition (as controls). Results: Before parenteral nutrition,the serum TBA, CG and HA levels in full-term neonates were not significantly different from those in the controls (P>0.05). After parenteral nutrition,serum levels were significantly higher than those before parenteral nutrition (P<0.01). The levels in pre-term neonates were significantly higher after parenteral nutrition than those in full-term neonates (P<0.05). Conclusion: Long term parenteral nutrition might be harmful to hepatic and gall bladder function in neonates especially in premature ones. (authors)
Chand, K K; Henshaw, W D; Lundquist, K A; Singer, M A
CgWind is a high-fidelity large eddy simulation (LES) tool designed to meet the modeling needs of wind turbine and wind park engineers. This tool combines several advanced computational technologies in order to model accurately the complex and dynamic nature of wind energy applications. The composite grid approach provides high-quality structured grids for the efficient implementation of high-order accurate discretizations of the incompressible Navier-Stokes equations. Composite grids also provide a natural mechanism for modeling bodies in relative motion and complex geometry. Advanced algorithms such as matrix-free multigrid, compact discretizations and approximate factorization will allow CgWind to perform highly resolved calculations efficiently on a wide class of computing resources. Also in development are nonlinear LES subgrid-scale models required to simulate the many interacting scales present in large wind turbine applications. This paper outlines our approach, the current status of CgWind and future development plans.
Rørbye, C; Nørgaard, M; Nilas, Lisbeth
thickness by ultrasonography was performed on day 15 after induction of medical abortion. Failures diagnosed after day 15 and within 15 weeks were identified and classified as late failures. All interventions in this group were due to bleeding problems. The predictive values of different absolute and...... relative beta-hCG levels and different endometrial thickness were analysed. RESULTS: Of all failures in the study period, 65% (32) were diagnosed after the 2 week follow-up, and categorized as late failures. The absolute and the relative beta-hCG values on days 8 and 15 as well as the endometrial thickness......BACKGROUND: Surgical treatment of failed medical abortion may be performed several weeks after initiation of the abortion. There are no recognized methods for early identification of these late failures. We assessed the prognostic values of beta-hCG and ultrasonography in predicting late failure...
Full Text Available Pronunciation practice system based on personalized Computer Graphics: CG animation of mouth movement model is proposed. The system enables a learner to practice pronunciation by looking at personalized CG animations of mouth movement model , and allows him/her to compare them with his/her own mouth movements. In order to evaluate the effectiveness of the system by using personalized CG animation of mouth movement model, Japanese vowel and consonant sounds were read by 8 infants before and after practicing with the proposed system, and their pronunciations were examined. Remarkable improvement on their pronunciations is confirmed through a comparison to their pronunciation without the proposed system based on identification test by subjective basis.
Although the glycoprotein hormones lutropin (LH), follitropin (FSH), and thyrotropin (TSH) bind to different receptors, each contains an identical alpha subunit. Specificity is somehow endowed by theta subunits which are distinct for each hormone. Human choriogonadotropin (hCG) is a natural LH analog that contains a beta subunit nearly identical to that of LH. The roles of these subunits in the recognition and high affinity binding of hCG to receptor was examined. Polyclonal and monoclonal antibodies specific for the individual subunits of hCG were used to probe the hormone-receptor interaction. Conformation-specific and sequence-specific antibodies were examined for their abilities to bind Triton X-100-solubilized 125I-hCG-receptor complex and to inhibit hormone binding to crude rat ovarian membranes containing receptor. Even though the immunoreactive sites are not located on the receptor binding surface of the beta subunit, most, but not all, of these polyclonal and monoclonal antibodies were able to inhibit 125I-hCG binding to receptor. Although the inhibition of binding may be due to steric interference due to the size of the antibody molecules, a two-step model for hCG binding to receptor is presented that also explains these results. In this model, the beta subunit initially binds with the receptor with a highly specific but low affinity interaction. This activates a site for the high affinity binding of the alpha subunit and stabilization of the complex. This is an attractive model as it may be applied to other glycoprotein hormones sharing an alpha subunit
Although the glycoprotein hormones lutropin (LH), follitropin (FSH), and thyrotropin (TSH) bind to different receptors, each contains an identical alpha subunit. Specificity is somehow endowed by theta subunits which are distinct for each hormone. Human choriogonadotropin (hCG) is a natural LH analog that contains a beta subunit nearly identical to that of LH. The roles of these subunits in the recognition and high affinity binding of hCG to receptor was examined. Polyclonal and monoclonal antibodies specific for the individual subunits of hCG were used to probe the hormone-receptor interaction. Conformation-specific and sequence-specific antibodies were examined for their abilities to bind Triton X-100-solubilized /sup 125/I-hCG-receptor complex and to inhibit hormone binding to crude rat ovarian membranes containing receptor. Even though the immunoreactive sites are not located on the receptor binding surface of the beta subunit, most, but not all, of these polyclonal and monoclonal antibodies were able to inhibit /sup 125/I-hCG binding to receptor. Although the inhibition of binding may be due to steric interference due to the size of the antibody molecules, a two-step model for hCG binding to receptor is presented that also explains these results. In this model, the beta subunit initially binds with the receptor with a highly specific but low affinity interaction. This activates a site for the high affinity binding of the alpha subunit and stabilization of the complex. This is an attractive model as it may be applied to other glycoprotein hormones sharing an alpha subunit.
Chedane, Carole; Puissant, Hugues; Weil, Dominique; Rouleau, Stéphanie; Coutant, Régis
Background An increase in cryptorchidism has been reported in many countries. One mechanism could be low fetal testosterone production possibly secondary to altered placental human chorionic gonadotrophin (hCG) release. Our Objective was to compare hCG values from maternal blood between boys with cryptorchidism and normal boys. Methods Total hCG and α-fetoprotein (AFP) values [12–16 weeks of gestation; from the double test for Down syndrome screening) were compared between cases of cryptorchi...
Kim, Jinyoung Serena; Walter, Frederick M.; Wolk, Scott J.
We present photometric and spectroscopic results for the low mass pre-main sequence (PMS) stars with spectral types K - M in the cometary globule (CG) 30/31/38 complex. We obtained multi-object high resolution spectra for the targets selected as possible PMS stars from multi-wavelength photometry. We identified 11 PMS stars brighter than V = 16.5 with ages < 5 Myr at a distance of approximately 200 pc. The spatial distribution of the PMS stars, CG clouds, and ionizing sources (O stars and sup...
L. Zicarelli; R. Di Palo; A. Balestrieri; C. Grassi; M. Rendina; Vecchio, D; G. Campanile
The aim of the present study was to examine whether treatment with a GnRH agonist or hCG in pregnant buffaloes on Day 25 after AI induce ovulation and increased P4 concentrations. The trial was carried out on 98 pluriparous buffaloes (DIM = 163 ± 75 days) diagnosed pregnant by ultrasound on day 25 after AI, and randomly assigned in two treatment groups GnRH (12 μg of Buserelin Acetate i.m) and hCG (1500 I.U. i.m.) after measurements of follicular diameter and evaluated ovulation. Mi...
Deconvolution problem is a main topic in signal processing. Many practical applications are re-quired to solve deconvolution problems. An important example is image reconstruction. Usually, researcherslike to use regularization method to deal with this problem. But the cost of computation is high due to thefact that direct methods are used. This paper develops a trust region-cg method, a kind of iterative methodsto solve this kind of problem. The regularity of the method is proved. Based on the special structure of thediscrete matrix, FFT can be used for calculation. Hence combining trust region-cg method with FFT is suitablefor solving large scale problems in signal processing.
Gao, Tie-Jun; Feng, Tai-Fu; Chen, Jian-Bin
In this work, we calculate the top quark rare decays t-cgamma and t-cg in the framework where the standard model is embedded in a warped extra dimension with the custodial symmetry SU(3)c*SU(2)L*SU(2)R*U(1)x*PLR. Adopting reasonable assumptions on the parameter space,we numerically find the branching ratios of t-cgamma exceeding 10^-6 and that of t-cg exceeding 10^-5 respectively, which can be detected in near future.
Martikainen Hannu; Tapanainen Juha S; Järvelä Ilkka Y
Abstract Background In natural cycles, women conceive when intercourse takes place during a six-day period ending on the day of ovulation. The current practice in intrauterine insemination (IUI) cycles is to perform the IUI 24-36 hours after the hCG administration, when the ovulation is already imminent. In this study hCG was administered after the IUI, which more closely resembles the fertilisation process in natural cycles. Methods All the IUIs performed since the beginning of 2007 were ana...
U radu je prikazano automatizirano praćenje rada relativnih gravimetara Scintrex Autograv CG-3M primjenom aplikacije Pragra, izrađene u programskom paketu MATLAB. Scintrex Autograv CG-3M je moderni relativni gravimetar. Nakon postavljanja gravimetra na točku, horizontiranja i pokretanja mjerenja, mjerni proces je potpuno automatiziran. Mjerni signal se već u gravimetru, na temelju definirane kalibracijske funkcije, pretvara u očitanja ubrzanja sile teže na koje se primjenjuju instrumentalne k...
Kanamori, Takashi; Masaki, Yoshiaki; Mizuta, Masahiro; Tsunoda, Hirosuke; Ohkubo, Akihiro; Sekine, Mitsuo; Seio, Kohji
We have previously reported DNA triplexes containing the unnatural base triad G-PPI·C3, in which PPI is an indole-fused cytosine derivative incorporated into DNA duplexes and C3 is an abasic site in triplex-forming oligonucleotides (TFOs) introduced by a propylene linker. In this study, we developed a new unnatural base triad A-ψ·C(R1) where ψ and C(R1) are base moieties 2'-deoxypseudouridine and 5-substituted deoxycytidine, respectively. We examined several electron-withdrawing substituents for R1 and found that 5-bromocytosine (C(Br)) could selectively recognize ψ. In addition, we developed a new PPI derivative, PPI(Me), having a methyl group on the indole ring in order to achieve selective triplex formation between DNA duplexes incorporating various Watson-Crick base pairs, such as T-A, C-G, A-ψ, and G-PPI(Me), and TFOs containing T, C, C(Br), and C3. We studied the selective triplex formation between these duplexes and TFOs using UV-melting and gel mobility shift assays. PMID:22146807
The aim of this study was to explore the value of serum cholylglycine (CG) in the diagnosis for intrahepatic cholestasis of pregnancy (ICP), in healthy pregnant and ICP women of Han and minority nationalities in Xinjiang. CG was measured by RIA. ALT was also measured and was analyzed comparatively with CG. The results showed that there was no significant difference between serum CG content of healthy Han women at second trimester pregnancy and that of healthy minority women at same period of pregnancy (P>0.05). However, serum CG content of minority women at third trimester pregnancy was significantly higher than that of Han women at same period of pregnancy (P<0.05). Serum CG content and ALT level of ICP patients from various nationalities were all higher than those of healthy pregnant women at the second and third trimester (P<0.01). Serum CG content and ALT level of minority ICP patients were significantly higher than those of Han ICP patients (P<0.01). Determination of serum CG should be taken as a routine procedure for pregnant women, for it is very important for timely diagnosis and treatment of ICP patients. At the same time, attention should be paid to monitoring serum CG level in minority pregnant women for the sake of strengthening health care work for minority pregnant women at perinatal period
Qi wang; Chengyi Yang; Huaxiang Wang; Ziqiang Cui; Zhentao Gao
Electrical capacitance tomography (ECT) is a promising technique for multi-phase flow measurement due to its high speed,low cost and non-intrusive sensing.Image reconstruction for ECT is an inverse problem of finding the permittivity distribution of an object by measuring the electrical capacitances between sets of electrodes placed around its periphery.The conjugate gradient (CG) method is a popular image reconstruction method for ECT,in spite of its low convergence rate.In this paper,an advanced version of the CG method,the projected CG method,is used for image reconstruction of an ECT system.The solution space is projected into the Krylov subspace and the inverse problem is solved by the CG method in a low-dimensional specific subspace.Both static and dynamic experiments were carried out for gas-solid two-phase flows.The flow regimes are identified using the reconstructed images obtained with the projected CG method.The results obtained indicate that the projected CG method improves the quality of reconstructed images and dramatically reduces computation time,as compared to the traditional sensitivity,Landweber,and CG methods.Furthermore,the projected CG method was also used to estimate the important parameters of the pneumatic conveying process,such as the volume concentration,flow velocity and mass flow rate of the solid phase.Therefore,the projected CG method is considered suitable for online gas-solid two-phase flow measurement.
Xie, Hang; Gursel, Ihsan; Ivins, Bruce E.; Singh, Manmohan; O'Hagan, Derek T.; Ulmer, Jeffrey B.; Klinman, Dennis M.
To reduce the biothreat posed by anthrax, efforts are under way to improve the protection afforded by vaccination. This work examines the ability of immunostimulatory CpG oligodeoxynucleotides (ODN) adsorbed onto cationic polylactide-co-glycolide (PLG) microparticles (CpG ODN-PLG) to accelerate and boost the protective immunity elicited by Anthrax Vaccine Adsorbed (AVA, the licensed human anthrax vaccine). The results indicate that coadministering CpG ODN-PLG with AVA induces a stronger and f...
Cazzamali, Giuseppe; Torp, Malene; Hauser, Frank;
The database from the Drosophila Genome Project contains a gene, CG9918, annotated to code for a G protein-coupled receptor. We cloned the cDNA of this gene and functionally expressed it in Chinese hamster ovary cells. We tested a library of about 25 Drosophila and other insect neuropeptides, and...
Duintjer Tebbens, Jurjen; Meurant, G.
2014. s. 26-26. [IMA International Conference on Numerical Linear Algebra and Optimisation /4./. 03.09.2014-05.09.2014, Birmingham] Institutional support: RVO:67985807 Keywords : prescribed convergence * Bi-Lanczos process * Bi-CG method Subject RIV: BA - General Mathematics
Kava, Hieronimus W; Galea, Anne M; Md Jamil, Farhana; Feng, Yue; Murray, Vincent
In this study, the DNA sequence specificity of four DNA-targeted 9-aminoacridine carboxamide Pt complexes was compared with cisplatin, using two specially constructed plasmid templates. One plasmid contained 5'-CG and 5'-GA insert sequences while the other plasmid contained a G-rich transferrin receptor gene promoter insert sequence. The damage profiles of each compound on the different DNA templates were quantified via a polymerase stop assay with fluorescently labelled primers and capillary electrophoresis. With the plasmid that contained 5'-CG and 5'-GA dinucleotides, the four 9-aminoacridine carboxamide Pt complexes produced distinctly different damage profiles as compared with cisplatin. These 9-aminoacridine complexes had greatly increased levels of DNA damage at CG and GA dinucleotides as compared with cisplatin. It was shown that the presence of a CG or GA dinucleotide was sufficient to reveal the altered DNA sequence selectivity of the 9-aminoacridine carboxamide Pt analogues. The DNA sequence specificity of the Pt complexes was also found to be similarly altered utilising the transferrin receptor DNA sequence. PMID:24827388
Becker, Michael; Krämer, Reinhard
Corynebacterium glutamicum is used in microbial biotechnology for the production of amino acids, e.g., glutamate and lysine. Excretion of glutamate into the surrounding medium under production conditions is mediated by MscCG, an MscS-type mechanosensitive channel. In difference to most other MscS-type channel proteins, MscCG carries, in addition to the N-terminal pore domain, a long C-terminal domain that amounts to about half of the size of the protein and harbors an additional transmembrane segment. Here we study the impact of the C-terminal domain on both functions of MscCG as mechanosensitive channel and as glutamate exporter. Sequential truncations of the C-terminal domain were applied, as well as deletion of particular subdomains, replacement of these segments by other amino acid sequences, and sequence randomization. Several parameters of cell physiology and bioenergetics of the obtained mutants related to both glutamate excretion and response to osmotic stress were quantified. All three subdomains of the C-terminal domain, i.e., the periplasmic loop, the fourth transmembrane segment, and the cytoplasmic loop, proved to be of core significance for MscCG function, in particular for glutamate excretion. PMID:26033538
Objective To explore the relationship between the lipoprotein lipase(LPL)-447C/G gene polymorphism and cerebral infarction in the elderly. Methods This was a case-control study,which enrolled 206 cases with cerebral infarction in the elderly and 203 elderly
Full Text Available Abstract Background Recently, it has been demonstrated that, in patients down-regulated by GnRH analogues (GnRHa, a short-term pre-treatment with recombinant LH (rLH, prior to recombinant FSH (rFSH administration, increases the number of small antral follicle prior to FSH stimulation and the yield of normally fertilized embryos. However, no data exist in the literature regarding the potential beneficial effect of "hCG priming" in controlled ovarian hyperstimulation (COH through a long GnRH-a protocol, which binds the same receptor (LH/hCGR, though it is a much more potent compared to LH. The primary aims of this study were to assess the effect of short-term pre-rFSH administration of hCG in women entering an ICSI treatment cycle on follicular development, quality of oocytes and early embryo development. The secondary endpoints were to record the effects on endometrial quality and pregnancy rate. Methods Patients with a history of at least one previous unsuccessful ICSI cycle were randomly assigned into two groups to receive treatment with either a long protocol with rFSH (control group or a long protocol with rFSH and pre-treatment with hCG (hCG group. In particular, in the latter group, a fixed 7 days course of 200 IU/day hCG was administered as soon as pituitary desensitization was confirmed. Results The mean number of oocytes retrieved was not significantly different between the two treatment groups, although the percentage of mature oocytes tended to be higher but not significantly different in hCG-treated patients. The percentage of patients with more than one grade 3 embryos was higher in the pre-treatment group, which also showed a higher pregnancy rate. Conclusion All the above clinical observations, in conjunction with previous data, suggest a point towards a beneficial "hCG priming" effect in controlled ovarian hyperstimulation through a long GnRH-a down-regulation protocol, particularly in patients with previous ART failures.
Jansson, Christer; Sun, Chuanxin; Ghebramedhin, Haile; Hoglund, Anna-Stina; Jansson, Christer
Antisense oligodeoxynucleotide (ODN) inhibition emerges as an effective means for probing gene function in plant cells. Employing this method we have established the importance of the SUSIBA2 transcription factor for regulation of starch synthesis in barley endosperm, and arrived at a model for the role of the SUSIBAs in sugar signaling and source-sink commutation during cereal endosperm development. In this addendum we provide additional data demonstrating the suitability of the antisense ODN technology in studies on starch branching enzyme activities in barley leaves. We also comment on the mechanism for ODN uptake in plant cells. Antisense ODNs are short (12-25 nt-long) stretches of single-stranded ODNs that hybridize to the cognate mRNA in a sequence-specific manner, thereby inhibiting gene expression. They are naturally occurring in both prokaryotes and eukaryotes where they partake in gene regulation and defense against viral infection. The mechanisms for antisense ODN inhibition are not fully understood but it is generally considered that the ODN either sterically interferes with translation or promotes transcript degradation by RNase H activation. The earliest indication of the usefulness of antisense ODN technology for the purposes of molecular biology and medical therapy was the demonstration in 1978 that synthetic ODNs complementary to Raos sarcoma virus could inhibit virus replication in tissue cultures of chick embryo fibroblasts. Since then the antisense ODN technology has been widely used in animal sciences and as an important emerging therapeutic approach in clinical medicine. However, antisense ODN inhibition has been an under-exploited strategy for plant tissues, although the prospects for plant cells in suspension cultures to take up single-stranded ODNs was reported over a decade ago. In 2001, two reports from Malho and coworker demonstrated the use of cationic-complexed antisense ODNs to suppress expression of genes encoding pollen
Christensen, T B; Engbaek, F; Marqversen, J;
days (3.6-6.8 days), whereas four out of five patients not achieving long-term remission had half-lives shorter than 3 days. After the third treatment cycle, eight patients who achieved long-term remission had hCG half-lives longer than 3 days (7.4-17.0 days). In these patients, the plasma...... disappearance of [125I]hCG was equivalent to that of hCG. Thus, the slow decline of hCG represented a slow plasma disappearance rather than a hCG production from vital tumour cells and could, consequently, not be used to select patients for additional or intensified chemotherapy. The concept of a fixed half...
Liu, Zheng-Wei; Stancliffe, Richard J.
The nature of the progenitors of Type Ia supernovae (SNe Ia) is not yet fully understood. In the single-degenerate (SD) scenario, the collision of the SN ejecta with its companion star is expected to produce detectable ultraviolet emission in the first few days after the SN explosion within certain viewing angles. It was recently found that the B - V colour of the nearby SN Ia SN 2012cg at about 16 d before the maximum B-band brightness was about 0.2 mag bluer than those of other normal SNe Ia, which was reported as the first evidence for excess blue light from the interaction of normal SN Ia ejecta with its companion star. In this work, we compare current observations for SN 2012cg from its pre-explosion phase to the late-time nebular phase with theoretical predictions from binary evolution and population synthesis calculations for a variety of popular progenitor scenarios. We find that a main-sequence donor or a carbon-oxygen white dwarf donor binary system is more likely to be the progenitor of SN 2012cg. However, both scenarios also predict properties which are in contradiction to the observed features of this system. Taking both theoretical and observational uncertainties into account, we suggest that it might be too early to conclude that SN 2012cg was produced from an explosion of a Chandrasekhar-mass white dwarf in the SD scenario. Future observations and improved detailed theoretical modelling are still required to place a more stringent constraint on the progenitor of SN 2012cg.
Riviere, Guillaume; Fellous, Alexandre; Franco, Alban; Bernay, Benoit; Favrel, Pascal
Angiotensin-converting enzyme (ACE) is a highly conserved metallopeptidase. In mammals, the somatic isoform governs blood pressure whereas the germinal isoform (tACE) is required for fertility. In Ecdysozoans, ACE-like enzymes are implicated in reproduction. Despite ACE orthologues being present from bacteria to humans, their function(s) remain(s) unknown in distant organisms such as Lophotrochozoans. In silico analysis of an oyster (Crassostrea gigas) EST library suggested the presence of an ACE orthologue in molluscs. Primer walking and 5'-RACE revealed that the 1.9 kb cDNA encodes CgACE, a 632 amino acid protein displaying a conserved single active site and a putative C-terminal transmembrane anchor, thus resembling human tACE, as supported by molecular modelling. FRET activity assays and Maldi-TOF spectrometry indicated that CgACE is a functional dipeptidyl-carboxypeptidase which is active on Angiotensin I and sensitive to ACE inhibitors and chloride ion concentration. Immunocytochemistry revealed that, as its human counterpart, recombinant CgACE is synthesised as a transmembrane enzyme. RT-qPCR, in-situ hybridization and immunohistochemistry shed light on a tissue, and development stage, specific expression pattern for CgACE, which is increased in the gonad during spermatogenesis. The use of ACE inhibitors in vivo indicates that the dipeptidase activity of CgACE is crucial for the oyster fertilization. Our study demonstrates that a transmembrane active ACE is present in the oyster Crassostrea gigas, and for the first time ascribes a functional role for ACE in Lophotrochozoans. Its biological function in reproduction is conserved from molluscs to humans, a finding of particular evolutionary interest especially since oysters represent the most important aquaculture resource worldwide. PMID:22174750
Full Text Available Angiotensin-converting enzyme (ACE is a highly conserved metallopeptidase. In mammals, the somatic isoform governs blood pressure whereas the germinal isoform (tACE is required for fertility. In Ecdysozoans, ACE-like enzymes are implicated in reproduction. Despite ACE orthologues being present from bacteria to humans, their function(s remain(s unknown in distant organisms such as Lophotrochozoans. In silico analysis of an oyster (Crassostrea gigas EST library suggested the presence of an ACE orthologue in molluscs. Primer walking and 5'-RACE revealed that the 1.9 kb cDNA encodes CgACE, a 632 amino acid protein displaying a conserved single active site and a putative C-terminal transmembrane anchor, thus resembling human tACE, as supported by molecular modelling. FRET activity assays and Maldi-TOF spectrometry indicated that CgACE is a functional dipeptidyl-carboxypeptidase which is active on Angiotensin I and sensitive to ACE inhibitors and chloride ion concentration. Immunocytochemistry revealed that, as its human counterpart, recombinant CgACE is synthesised as a transmembrane enzyme. RT-qPCR, in-situ hybridization and immunohistochemistry shed light on a tissue, and development stage, specific expression pattern for CgACE, which is increased in the gonad during spermatogenesis. The use of ACE inhibitors in vivo indicates that the dipeptidase activity of CgACE is crucial for the oyster fertilization. Our study demonstrates that a transmembrane active ACE is present in the oyster Crassostrea gigas, and for the first time ascribes a functional role for ACE in Lophotrochozoans. Its biological function in reproduction is conserved from molluscs to humans, a finding of particular evolutionary interest especially since oysters represent the most important aquaculture resource worldwide.
Endo, Akira; Kim, Eunjoo; Yamaguchi, Yasuhiro [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment
A Monte Carlo code SCINFUL has been utilized for calculating response functions of organic scintillators for high-energy neutron spectroscopy. However, the applicability of SCINFUL is limited to the calculations for cylindrical NE213 and NE110 scintillators. In the present study, SCINFUL-CG was developed by introducing a geometry specifying function and high-energy neutron cross section data into SCINFUL. The geometry package MARS-CG, the extended version of the CG (Combinatorial Geometry), was programmed into SCINFUL-CG to express various geometries of detectors. Neutron spectra in the regions specified by the CG can be evaluated by the track length estimator. The cross section data of silicon, oxygen and aluminum for neutron transport calculation were incorporated up to 100 MeV using the data of LA150 library. Validity of SCINFUL-CG was examined by comparing calculated results with those by SCINFUL and MCNP and experimental data measured using high-energy neutron fields. SCINFUL-CG can be used for the calculations of the response functions and neutron spectra in the organic scintillators in various shapes. The computer code will be applicable to the designs of high-energy neutron spectrometers and neutron monitors using the organic scintillators. The present report describes the new features of SCINFUL-CG and explains how to use the code. (author)
Full Text Available Huijie Zhang,1 Shini Feng,1 Ting Yan,1 Chunyi Zhi,2 Xiao-Dong Gao,1 Nobutaka Hanagata3,41The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, People’s Republic of China; 2Department of Physics and Materials Science, City University of Hong Kong, Kowlong, Hong Kong SAR, People’s Republic of China; 3Biomaterials Unit, International Center for Materials Nanoarchitectonics, National Institute for Materials Science, Ibaraki, Japan; 4Nanotechnology Innovation Station, National Institute for Materials Science, Ibaraki, JapanAbstract: CpG oligodeoxynucleotides (ODNs stimulate innate and adaptive immune responses. Thus, these molecules are promising therapeutic agents and vaccine adjuvants against various diseases. In this study, we developed a novel CpG ODNs delivery system based on polyethyleneimine (PEI-functionalized boron nitride nanospheres (BNNS. PEI was coated on the surface of BNNS via electrostatic interactions. The prepared BNNS–PEI complexes had positive zeta potential and exhibited enhanced dispersity and stability in aqueous solution. In vitro cytotoxicity assays revealed that the BNNS–PEI complexes with concentrations up to 100 µg/mL exhibited no obvious cytotoxicity. Furthermore, the positively charged surface of the BNNS–PEI complexes greatly improved the loading capacity and cellular uptake efficiency of CpG ODNs. Class B CpG ODNs loaded on the BNNS–PEI complexes enhanced the production of interleukin-6 and tumor necrosis factor-α from peripheral blood mononuclear cells compared with CpG ODNs directly loaded on BNNS. Contrary to the free CpG ODNs or CpG ODNs directly loaded on BNNS, class B CpG ODNs loaded on the BNNS–PEI complexes induced interferon-α simultaneously. PEI coating may have changed the physical form of class B CpG ODNs on BNNS, which further affected their interaction with Toll-like receptor 9 and induced interferon
The quantities c(G), q(G) and p(G) for finite groups were defined by H.Behravesh. In this article, these quantities for the alternating group An and the symmetric group Sn are calculated. It is shown thatc(G) = q(G) = p(G) = n,when G = An or Sn.
Li, Xiu-Qing; Du, Donglei
C+G content (GC content or G+C content) is known to be correlated with genome/chromosome size in bacteria but the relationship for other kingdoms remains unclear. This study analyzed genome size, chromosome size, and base composition in most of the available sequenced genomes in various kingdoms. Genome size tends to increase during evolution in plants and animals, and the same is likely true for bacteria. The genomic C+G contents were found to vary greatly in microorganisms but were quite similar within each animal or plant subkingdom. In animals and plants, the C+G contents are ranked as follows: monocot plants>mammals>non-mammalian animals>dicot plants. The variation in C+G content between chromosomes within species is greater in animals than in plants. The correlation between average chromosome C+G content and chromosome length was found to be positive in Proteobacteria, Actinobacteria (but not in other analyzed bacterial phyla), Ascomycota fungi, and likely also in some plants; negative in some animals, insignificant in two protist phyla, and likely very weak in Archaea. Clearly, correlations between C+G content and chromosome size can be positive, negative, or not significant depending on the kingdoms/groups or species. Different phyla or species exhibit different patterns of correlation between chromosome-size and C+G content. Most chromosomes within a species have a similar pattern of variation in C+G content but outliers are common. The data presented in this study suggest that the C+G content is under genetic control by both trans- and cis- factors and that the correlation between C+G content and chromosome length can be positive, negative, or not significant in different phyla. PMID:24551092
Merzetti, E M; Staveley, B E
F-box proteins act as the protein recognition component of the Skp-Cul-F-box class of ubiquitin ligases. Two members of a gene sub-family encoding these proteins, FBXO7 and FBXO32, have been implicated in the onset and progression of degenerative disease. FBXO7 is responsible for rare genetic forms of Parkinson disease, while FBXO32 has been implicated in muscle wasting. The third gene in this family, FBXO9, is related to growth signaling, but the role of this gene in degenerative disease pathways has not been thoroughly investigated. Characterizing the putative Drosophila melanogaster homologue of this gene, CG5961, enables modeling and analysis of the consequence of targeted alteration of gene function and the effects on the overall health of the organism. Comparison of the protein domains of Homo sapiens FBXO9 and the putative D. melanogaster homologue CG5961 revealed a high degree of conservation between the protein domains. Directed expression of CG5961 (via CG5961(EP)) and inhibition of CG5961 (through a stable RNAi transgene) in the developing D. melanogaster eye caused abnormalities in adult structures (ommatidia and inter-ommatidial bristles). Directed expression of either CG5961 or CG5961-RNAi in the dopaminergic neurons led to a reduced lifespan compared to that in lacZ controls. We showed that protein structures of CG5961 and FBXO9 are highly similar and studied the effects of altered expression of CG5961 in neuron-rich tissues. Our results suggest that CG5961 activity is necessary for the proper formation of neuronal tissue and that targeted alteration of gene expression in dopaminergic neurons leads to a reduced lifespan. PMID:27173356
This book contains the following chapters: The Military and Alternative Security: New Missions for Stable Conventional Security; Technology and Alternative Security: A Cherished Myth Expires; Law and Alternative Security: Toward a Just World Peace; Politics and Alternative Security: Toward a More Democratic, Therefore More Peaceful, World; Economics and Alternative Security: Toward a Peacekeeping International Economy; Psychology and Alternative Security: Needs, Perceptions, and Misperceptions; Religion and Alternative Security: A Prophetic Vision; and Toward Post-Nuclear Global Security: An Overview
Gil, Lázaro; Marcos, Ernesto; Izquierdo, Alienys; Lazo, Laura; Valdés, Iris; Ambala, Peris; Ochola, Lucy; Hitler, Rikoi; Suzarte, Edith; Álvarez, Mayling; Kimiti, Prisilla; Ndung'u, James; Kariuki, Thomas; Guzmán, María Guadalupe; Guillén, Gerardo; Hermida, Lisset
Previously, we reported the ability of the chimeric protein DIIIC-2 (domain III of the dengue envelope protein fused to the capsid protein of dengue-2 virus), to induce immunity and protection in mice, when it is highly aggregated with a non-defined oligodeoxynucleotide (ODN) and adjuvanted in alum. In this work, three different defined ODNs were studied as aggregating agents. Our results suggest that the nature of the ODN influences the capacity of protein DIIIC-2 to activate cell-mediated immunity in mice. Consequently, the ODN 39M was selected to perform further experiments in mice and nonhuman primates. Mice receiving the preparation 39M-DIIIC-2 were solidly protected against dengue virus (DENV) challenge. Moreover, monkeys immunized with the same preparation developed neutralizing antibodies, as measured by four different neutralization tests varying the virus strains and the cell lines used. Two of the immunized monkeys were completely protected against challenge, whereas the third animal had a single day of low-titer viremia. This is the first work describing the induction of short-term protection in monkeys by a formulation that is suitable for human use combining a recombinant protein from DENV with alum. PMID:25178969
Yu Fan; Shu Zheng; Ze-Feng Xu; Jia-Yi Ding
AIM: To investigate the effects of polo-like kinase-1 (PLK1) antisense phosphorothioate oligodeoxynucleotide (ASODN) on apoptosis and cell cycle of human colon cancer cell line 5W480.METHODS: After SW480 colon cancer cells were transfected with PLK1 ASODN, Northern and Western blot analyses were used to examine PLK1 gene expression in cancer cells. We studied apoptosis using terminal uridine deoxynucleotidyl nick end labeling. Apoptosis and cell cycle of SW480 cells were examined by fluorescence-activated cell sorter scan.RESULTS: The levels of PLK1 mRNA and protein were greatly inhibited by PLK1 ASODN in SW480 cancer cells transfected with PLK1 ASODN. Apoptosis index (AI) induced PLK1 ASODN in a time- and dose-dependent manner.Results from FLM showed that sub-2N DNA content of transfected cancer cells was significantly increased and arrested at G2/M compared with control groups.CONCLUSION: PLK1 ASODN can induce apoptosis of human colon cancer cell line SW480.
Yun-Feng Wang; Ke-Jian Guo; Bei-Ting Huang; Yong Liu; Xiao-Yun Tang; Jian-Jun Zhang; Qiang Xia
AIM: To investigate the effect of telomerase hTERT gene antisense oligonucleotide (hTERT-ASO) on proliferation and telomerase activity of pancreatic cancer cell line Bxpc-3.METHODS: MTT assay was used to detect the effect of different doses of hTERT-ASO on proliferation of Bxpc-3 cell for different times. To study the anti-tumor activity,the cells were divided into there groups: Control group (pancreatic cancer cell Bxpc-3); antisense oligonucleotide (hTERT-ASO) group; and nosense oligonucleotide group decorated with phosphorothioate. Telomerase activity was detected using TRAP-PCR-ELISA. Cell DNA distribution was examined using flow cytometry assay.Cell apoptosis was observed by transmission electron microscope in each group.RESULTS: After treatment with 6 mmol/L hTERTASO, cell proliferation was inhibited in dose- and timedependent manner. The telomerase activity decreased after treatment with hTERT-ASO for 72 h. Flow cytometry showed the cell number of G0/G1 phase increased from 2.7% to 14.7%, the cell number of S phase decreased from 72.7% to 51.0%, and a sub-G1 stage cell apoptosis peak appeared in front of G1 stage.CONCLUSION: Telomerase antisense oligodeoxynucleotide can inhibit the proliferation of pancreatic cancer cell line Bxpc-3 and decrease the telomerase activity and increase cell apoptosis rate in vitro.
Weiner, George J.; Liu, Hsin-Ming; Wooldridge, James E.; Dahle, Christopher E.; Krieg, Arthur M.
Recent advances in our understanding of the immune response are allowing for the logical design of new approaches to cancer immunization. One area of interest is the development of new immune adjuvants. Immunostimulatory oligodeoxynucleotides containing the CpG motif (CpG ODN) can induce production of a wide variety of cytokines and activate B cells, monocytes, dendritic cells, and NK cells. Using the 38C13 B cell lymphoma model, we assessed whether CpG ODN can function as immune adjuvants in tumor antigen immunization. The idiotype served as the tumor antigen. Select CpG ODN were as effective as complete Freund's adjuvant at inducing an antigen-specific antibody response but were associated with less toxicity. These CpG ODN induced a higher titer of antigen-specific IgG2a than did complete Freund's adjuvant, suggesting an enhanced TH1 response. Mice immunized with CpG ODN as an adjuvant were protected from tumor challenge to a degree similar to that seen in mice immunized with complete Freund's adjuvant. We conclude that CpG ODN are effective as immune adjuvants and are attractive as part of a tumor immunization strategy.
Full Text Available DNA vaccines showed great promise in preclinical models of infectious and malignant diseases, but their potency was insufficient in clinical trials and is needed to be improved. In this study, we tested systemic administration of two conventional adjuvants, synthetic oligodeoxynucleotide carrying immunostimulatory CpG motifs (CpG-ODN and levamisole (LMS, and evaluated their effect on immune reactions induced by DNA vaccines delivered by a gene gun. DNA vaccination was directed either against the E7 oncoprotein of human papillomavirus type 16 or against the BCR-ABL1 oncoprotein characteristic for chronic myeloid leukemia. High doses of both adjuvants reduced activation of mouse splenic CD8+ T lymphocytes, but the overall antitumor effect was enhanced in both tumor models. High-dose CpG-ODN exhibited a superior adjuvant effect in comparison with any combination of CpG-ODN with LMS. In summary, our results demonstrate the benefit of combined therapy with gene-gun-delivered antitumor DNA vaccines and systemic administration of CpG-ODN or LMS.
CHUN YAN HE; QING LIANG LIU
CpG oligodeoxynucleotides (CpG ODN) as adjuvant have been extensively studied in recent years. Phosphodiester CpG ODN (PO CpG ODN) can perfectly mimic bacterial DNA in enhancing immune response but are vulnerable to nucleases in vivo. This study aimed to evaluate the immunostimu latory potential and safety of phosphodiester CpG ODN encapsulated in nonphospholipid liposomes.BALB/c mice were immunized intramuscularly with different formulations of liposomes, CpG ODN and hepatitis B surface antigen (HBsAg). The results demonstrated that the encapsulated PO CpG ODN were protected against rapid degradation in vivo and retained their adjuvant activity. PO CpG ODN encapsulated with HBsAg in liposomes induced strong Th1-biased or Th1/Th2 mixed humoral immune response in mice with the magnitude similar to their phosphothioate equivalent in the same formulation.High IFN-gamma production induced by this formulation confirmed the generation of strong cellular immune response. Additionally, co-delivery of HBsAg and PO CpG ODN improved the immune response over that obtained with separate delivery. Safety experiment showed that liposome-encapsulaed PO CpG ODN and HBsAg caused mild systemic and moderate local adverse reaction. In conclusion, our data shows that PO CpG ODN encapsulated in liposomes fully exhibit their Th1-type adjuvant activity and act as a potential adjuvant for vaccines.
Full Text Available Human luteinizing hormone (hLH and chorionic gonadotropin (hCG act on the same receptor (LHCGR but it is not known whether they elicit the same cellular and molecular response. This study compares for the first time the activation of cell-signalling pathways and gene expression in response to hLH and hCG. Using recombinant hLH and recombinant hCG we evaluated the kinetics of cAMP production in COS-7 and hGL5 cells permanently expressing LHCGR (COS-7/LHCGR, hGL5/LHCGR, as well as cAMP, ERK1/2, AKT activation and progesterone production in primary human granulosa cells (hGLC. The expression of selected target genes was measured in the presence or absence of ERK- or AKT-pathways inhibitors. In COS-7/LHCGR cells, hCG is 5-fold more potent than hLH (cAMP ED(50: 107.1±14.3 pM and 530.0±51.2 pM, respectively. hLH maximal effect was significantly faster (10 minutes by hLH; 1 hour by hCG. In hGLC continuous exposure to equipotent doses of gonadotropins up to 36 hours revealed that intracellular cAMP production is oscillating and significantly higher by hCG versus hLH. Conversely, phospho-ERK1/2 and -AKT activation was more potent and sustained by hLH versus hCG. ERK1/2 and AKT inhibition removed the inhibitory effect on NRG1 (neuregulin expression by hLH but not by hCG; ERK1/2 inhibition significantly increased hLH- but not hCG-stimulated CYP19A1 (aromatase expression. We conclude that: i hCG is more potent on cAMP production, while hLH is more potent on ERK and AKT activation; ii hGLC respond to equipotent, constant hLH or hCG stimulation with a fluctuating cAMP production and progressive progesterone secretion; and iii the expression of hLH and hCG target genes partly involves the activation of different pathways depending on the ligand. Therefore, the LHCGR is able to differentiate the activity of hLH and hCG.
Fitzpatrick David G
Full Text Available Abstract Background We report the case of a patient with treated Stage Ia seminoma who was found to have an elevated beta human chorionic gonadotrophin (hCG on routine follow – up. This instigated restaging and could have lead to commencement of chemotherapy. Case presentation The patient was a bodybuilder, and following a negative metastatic work – up, admitted to injecting exogenous beta hCG. This was done to reduce withdrawal symptoms from androgen abuse. The patient remains well eight years post diagnosis. Conclusion This case highlights the need for surgical oncologists to conduct vigilant screening of young male patients with a history of testicular germ cell tumours and who may indulge in steroid abuse.
Encarnacao, L. M.; Chuang, Yung-Yu; Stork, Andre; Kasik, David; Rhyne, Theresa-Marie; Avila, Lisa; Kohlhammer, Jorn; LaViola, Joseph; Tory, Melanie; Dill, John; Domik, Gitta; Owen, G. Scott; Wong, Pak C.
With many new members joining the CG&A editorial board over the past year, and with a renewed commitment to not only document the state of the art in computer graphics research and applications but to anticipate and where possible foster future areas of scientific discourse and industrial practice, we asked editorial and advisory council members about where they see their fields of expertise going. The answers compiled here aren’t meant to be all encompassing or deterministic when it comes to the opportunities computer graphics and interactive visualization hold for the future. Instead, we aim to accomplish two things: give a more in-depth introduction of members of the editorial board to the CG&A readership and encourage cross-disciplinary discourse toward approaching, complementing, or disputing the visions laid out in this compilation.
Human chorionic gonadotropin (hCG) is a two chains glycoprotein hormone normally found in blood and urine, only during pregnancy. It is secreted by placental tissue, beginning with the primitive trophoblast, almost from the time of implantation. The kinetics of the interaction between the hCG antigen and two matched antibodies, one labelled with radioactive isotope 125I (tracer) and the other is unlabelled and attached to a solid support (tube) , are studied using, two-sites (sandwich) Immunoradiometric assay (IRMA) technique. A new method for determining the rate constants, using an advanced computer program (RKY) based on the Nelder-Mead optimisation principle is introduced. The rate constants, at three variable temperatures and three different antigen concentrations as well as the half time of exchange (t) were calculated
Dash, S.; Mishra, G. P.
The cylindrical gate tunnel FET (CG-TFET) is one of the potential candidates for future nano-technology, as it exhibit greater scaling capability and low subthreshold swing (SS) as compared to conventional MOSFET. In this paper, a new analytical approach is proposed to extract the gate dependent threshold voltage for CG-TFET. The potential distribution and electric field distribution in the cylindrical channel has been obtained using the 2-D Poisson's equation which in turn computes the shortest tunneling distance and tunneling current. The threshold voltage is extracted using peak transconductance change method based on the saturation of tunneling barrier width. The impact of scaling of effective oxide thickness, cylindrical pillar diameter and gate length on the threshold voltage has been investigated. The consistency of the proposed model is validated with the TCAD simulated results. The present model can be a handful for the study of switching behavior of TFET.
Aherne, Noel J
BACKGROUND: We report the case of a patient with treated Stage Ia seminoma who was found to have an elevated beta human chorionic gonadotrophin (hCG) on routine follow - up. This instigated restaging and could have lead to commencement of chemotherapy. CASE PRESENTATION: The patient was a bodybuilder, and following a negative metastatic work - up, admitted to injecting exogenous beta hCG. This was done to reduce withdrawal symptoms from androgen abuse. The patient remains well eight years post diagnosis. CONCLUSION: This case highlights the need for surgical oncologists to conduct vigilant screening of young male patients with a history of testicular germ cell tumours and who may indulge in steroid abuse.
Shadid, J.N.; Hutchinson, S.A.; Moffat, H.K. [Sandia National Labs., Albuquerque, NM (United States)
A parallel unstructured finite element (FE) implementation designed for message passing MIMD machines is described. This implementation employs automated problem partitioning algorithms for load balancing unstructured grids, a distributed sparse matrix representation of the global finite element equations and a parallel conjugate gradient (CG) solver. In this paper a number of issues related to the efficient implementation of parallel unstructured mesh applications are presented. These include the differences between structured and unstructured mesh parallel applications, major communication kernels for unstructured CG solvers, automatic mesh partitioning algorithms, and the influence of mesh partitioning metrics on parallel performance. Initial results are presented for example finite element (FE) heat transfer analysis applications on a 1024 processor nCUBE 2 hypercube. Results indicate over 95% scaled efficiencies are obtained for some large problems despite the required unstructured data communication.
Christensen, T. B.; Engbaek, F; Marqversen, J; Nielsen, S I; Kamby, C.; Maase, H von der
The rate of reduction in the concentration of serum human chorionic gonadotrophin (hCG) following chemotherapy for germ cell tumours may follow a complex pattern, with longer apparent half-life during later stages of chemotherapy, even in patients treated successfully. The commonly used half-life of less than 3 days for hCG to monitor the effect of chemotherapy in patients with germ cell tumours of the testis may represent too simple a model. 125I-labelled hCG was injected intravenously in 27...
Nakayama, Yoshitaka; Becker, Michael; Ebrahimian, Haleh; Konishi, Tomoyuki; Kawasaki, Hisashi; Krämer, Reinhard; Martinac, Boris
The mechanosensitive (MS) channel MscCG from the soil bacterium Corynebacterium glutamicum functions as a major glutamate exporter. MscCG belongs to a subfamily of the bacterial MscS-like channels, which play an important role in osmoregulation. To understand the structural and functional features of MscCG, we investigated the role of the carboxyl-terminal domain, whose relevance for the channel gating has been unknown. The chimeric channel MscS-(C-MscCG), which is a fusion protein between the carboxyl terminal domain of MscCG and the MscS channel, was examined by the patch clamp technique. We found that the chimeric channel exhibited MS channel activity in Escherichia coli spheroplasts characterized by a lower activation threshold and slow closing compared to MscS. The chimeric channel MscS-(C-MscCG) was successfully reconstituted into azolectin liposomes and exhibited gating hysteresis in a voltage-dependent manner, especially at high pipette voltages. Moreover, the channel remained open after releasing pipette pressure at membrane potentials physiologically relevant for C. glutamicum. This contribution to the gating hysteresis of the C-terminal domain of MscCG confers to the channel gating properties highly suitable for release of intracellular solutes. PMID:26494188
Full Text Available Yuvaraj Manoharan,1,* Qingmin Ji,2,* Tomohiko Yamazaki,2,3 Shanmugavel Chinnathambi,1 Song Chen,2,4 Ganesan Singaravelu,1 Jonathan P Hill,2 Katsuhiko Ariga,2,5 Nobutaka Hanagata3,6 1Department of Medical Physics, Anna University, Chennai, India; 2Research Center for Materials Nanoarchitectonics, National Institute for Materials Science, Tsukuba, Ibarak, 3Graduate School of Life Science, Hokkaido University, Kita-ku, Sapporo, 4JSPS Research Fellow, Chiyoda-ku, Tokyo, 5JST and CREST, National Institute for Materials Science, Tsukuba, Ibaraki, Japan; 6Nanotechnology Innovation Station, National Institute for Materials Science, Tsukuba, Ibaraki, Japan*These authors contributed equally to this workBackground: Class B CpG oligodeoxynucleotides primarily interact with Toll-like receptor 9 (TLR9 in B cells and enhance the immune system through induction of various interleukins including interleukin-6 in these immune cells. Although free class B CpG oligodeoxynucleotides do not induce interferon (IFN-α production, CpG oligodeoxynucleotide molecules have been reported to induce IFN-α when loaded onto nanoparticles. Here, we investigated the in vitro induction of IFN-α by a nanocarrier delivery system for class B CpG oligodeoxynucleotide molecules.Methods: For improving the capacity to load CpG oligodeoxynucleotide molecules, flake-shell SiO2 nanoparticles with a specific surface area approximately 83-fold higher than that of smooth-surfaced SiO2 nanoparticles were prepared by coating SiO2 nanoparticles with polyethyleneimine (PEI of three different number-average molecular weights (Mns 600, 1800, and 10,000 Da.Results: The capacity of the flake-shell SiO2 nanoparticles to load CpG oligodeoxynucleotides was observed to be 5.8-fold to 6.7-fold higher than that of smooth-surfaced SiO2 nanoparticles and was found to increase with an increase in the Mn of the PEI because the Mn contributed to the positive surface charge density of the nanoparticles. Further
Full Text Available The aim of the present study was to examine whether treatment with a GnRH agonist or hCG in pregnant buffaloes on Day 25 after AI induce ovulation and increased P4 concentrations. The trial was carried out on 98 pluriparous buffaloes (DIM = 163 ± 75 days diagnosed pregnant by ultrasound on day 25 after AI, and randomly assigned in two treatment groups GnRH (12 μg of Buserelin Acetate i.m and hCG (1500 I.U. i.m. after measurements of follicular diameter and evaluated ovulation. Milk samples were collected on Days 30 and 45 after AI, to assess P4 concentrations in whey. Differences between the follicular diameters of ovulation and P4 were tested by ANOVA. The incidences of animals which responded to the two treatments were analysed by Chi square test. The treatments on day 25 after AI induced ovulation respectively in 68.6% (GnRH and 57.4% (hCG of the buffaloes. No differences were found between diameter of follicle ovulated. Ovulation increased milk whey progesterone levels and reduced embryonic mortality in buffalo cows.
Cullum, J. [IBM Research Division, Yorktown Heights, NY (United States)
We study the convergence of GMRES/FOM and QMR/BiCG methods for solving nonsymmetric Ax=b. We prove that given the results of a BiCG computation on Ax=b, we can obtain a matrix B with the same eigenvalues as A and a vector c such that the residual norms generated by a FOM computation on Bx=c are identical to those generated by the BiCG computations. Using a unitary equivalence for each of these methods, we obtain test problems where we can easily vary certain spectral properties of the matrices. We use these test problems to study the effects of nonnormality on the convergence of GMRES and QMR, to study the effects of eigenvalue outliers on the convergence of QMR, and to compare the convergence of restarted GMRES, QMR, and BiCGSTAB across a family of normal and nonnormal problems. Our GMRES tests on nonnormal test matrices indicate that nonnormality can have unexpected effects upon the residual norm convergence, giving misleading indications of superior convergence over QMR when the error norms for GMRES are not significantly different from those for QMR. Our QMR tests indicate that the convergence of the QMR residual and error norms is influenced predominantly by small and large eigenvalue outliers and by the character, real, complex, or nearly real, of the outliers and the other eigenvalues. In our comparison tests QMR outperformed GMRES(10) and GMRES(20) on both the normal and nonnormal test matrices.
The nature of the progenitors of SNe Ia is not yet fully understood. In the single-degenerate (SD) scenario, the collision of the SN ejecta with its companion star is expected to produce detectable UV emission in the first few days after the SN explosion within certain viewing angles. It was recently found that the $B-V$ colour of the nearby SN Ia SN 2012cg at about sixteen days before the maximum $B$-band brightness was about 0.2 mag bluer than those of other normal SNe Ia, which was reported as the first evidence for excess blue light from the interaction of normal SN Ia ejecta with its companion star. In this work, we compare current observations for SN 2012cg from its pre-explosion phase to the late-time nebular phase with theoretical predictions from binary evolution and population synthesis calculations for a variety of popular progenitor scenarios. We find that a main-sequence donor or a carbon-oxygen white dwarf donor binary system is more likely to be the progenitor of SN 2012cg. However, both scenar...
Cunha, Rodrigo Casquero; Andreotti, Renato; Leite, Fábio Pereira Leivas
The cattle tick Rhipicephalus (Boophilus) microplus is responsible for great economic losses. It is mainly controlled chemically, with limitations regarding development of resistance to the chemicals. Vaccines may help control this parasite, thereby reducing tick pesticide use. In this light, we performed subcloning of the gene of the protein Bm86-GC, the homologue protein that currently forms the basis of vaccines (Gavac(TM) and TickGard(PLUS)) that have been developed against cattle ticks. The subcloning was done in the pPIC9 expression vector, for transformation in the yeast Pichia pastoris. This protein was characterized by expression of the recombinant Mut+ strain, which expressed greater quantities of protein. The expressed protein (rBm86-CG) was recognized in the Western-blot assay using anti-Gavac, anti-TickGard, anti-larval extract and anti-rBm86-CG polyclonal sera. The serum produced in cattle vaccinated with the antigen CG rBm86 presented high antibody titers and recognized the native protein. The rBm86-GC has potential relevance as an immunogen for vaccine formulation against cattle ticks. PMID:21722483
Most of the computational work in the numerical simulation of fluid and heat flows in permeable media arises in the solution of large systems of linear equations. The simplest technique for solving such equations is by direct methods. However, because of large storage requirements and accumulation of roundoff errors, the application of direct solution techniques is limited, depending on matrix bandwidth, to systems of a few hundred to at most a few thousand simultaneous equations. T2CG1, a package of preconditioned conjugate gradient solvers, has been added to TOUGH2 to complement its direct solver and significantly increase the size of problems tractable on PCs. T2CG1 includes three different solvers: a Bi-Conjugate Gradient (BCG) solver, a Bi-Conjugate Gradient Squared (BCGS) solver, and a Generalized Minimum Residual (GMRES) solver. Results from six test problems with up to 30,000 equations show that T2CG1 (1) is significantly (and invariably) faster and requires far less memory than the MA28 direct solver, (2) it makes possible the solution of very large three-dimensional problems on PCs, and (3) that the BCGS solver is the fastest of the three in the tested problems. Sample problems are presented related to heat and fluid flow at Yucca Mountain and WIPP, environmental remediation by the Thermal Enhanced Vapor Extraction System, and geothermal resources
Wen, Deng-Tai; Zheng, Lan; Ni, Liu; Wang, Hui; Feng, Yue; Zhang, Min
The CG9940 gene, which encodes the NAD(+) synthase protein in Drosophila, is conserved in human, zebra fish, and mosquito. NAD(+) synthase is a homodimer, which catalyzes the final step in de novo nicotinamide adenine dinucleotide (NAD(+)) biosynthesis, an amide transfer from either ammonia or glutamine to nicotinic acid adenine dinucleotide (NaAD). Both the CG9940 and exercise are closely relative to NAD(+) level, and NAD(+) plays important roles not only in energy metabolism and mitochondrial functions but also in aging. In our study, the expression of CG9940 was changed by UAS/GAL4 system in Drosophila. Flies were trained by a training device. Cardiac function was analyzed by M-mode traces, climbing index was measured through negative geotaxis assay, and lifespan was measured via lifespan assays. The important new findings from our present study included the following: (1) the expression of the CG9940 could affect cardiac function, mobility, and lifespan in Drosophila. Over-expression of the CG9940 gene had positive effects on Drosophila, such as enhanced aging cardiac output, reduced heart failure, delayed age-related mobility decline, and prolonged lifespan, but lower-expression of the CG9940 had negative effects on them. (2) Different expressions of the CG9940 resulted in different influences on the adaptation of cardiac function, mobility, and lifespan to exercise in aging Drosophila. Both normal-expression and over-expression of the CG9940 resulted in positive influences on the adaptation of cardiac functions, mobility, and lifespan to exercise in aging Drosophila such as exercise slowed age-related decline of cardiac function, mobility and extent of lifespan in these flies, while lower-expression of the CG9940 led to negative impacts on the adaptation of mobility and lifespan to exercise in Drosophila. PMID:27448710
Pol, Olga; Valle, Lluís; Sánchez-Blázquez, Pilar; Garzón, Javier; Puig, Margarita M.
We have studied the effects of μ and δ opioids on intestinal function (permeability, PER; gastrointestinal transit, GIT), and their antagonism after the intracerebroventricular (i.c.v.) administration of specific antibodies (ABs) or antisense oligodeoxynucleotides (ODN) to μ-receptors (OR). Central versus peripheral site/s of action of subcutaneous (s.c.) μ-opioids, were also assessed.Male Swiss CD-1 mice were used. GIT was measured with charcoal and PER by the passage of 51Cr-EDTA from blood...
Kim, Do-Hyun; Sohn, Jung-Ho; Park, Hong-Jai; Lee, Jae-Hyun; Park, Jung-Won; Choi, Je-Min
Purpose CpG oligodeoxynucleotide (CpG-ODN), a TLR9 agonist, activates innate immunity and induces Th1 response. Although the immune modulatory effect of CpG-ODN has been extensively studied, its function in cockroach extract-induced allergic asthma has not been studied. Here, we investigated the inhibitory function of CpG-ODN in cockroach extract-induced asthma in mice with different treatment schemes. Methods Scheme 1: BALB/C mice were intra-nasally co-administered by cockroach extract and C...
Del Bufalo, D; Cucco, C.; C. Leonetti; Citro, G.; D'Agnano, I; M. Benassi; Geiser, T; Zon, G; Calabretta, B; Zupi, G.
We investigated the effect of c-myb antisense phosphorothioate oligodeoxynucleotides [(S)ODNs] and cisplatin (CDDP) combination on the human colon carcinoma cell line LoVo Dx both in vitro and in nude mice bearing LoVo Dx solid tumour. We show that antisense (S)ODN treatment decreases c-myb mRNA and protein expression, induces growth arrest in the G1 phase of the cell cycle, and inhibits cell proliferation. In vivo treatment with c-myb antisense (S)ODNs results in a reduction in tumour growth...
Chen, Wen-Tsen; Huang, Ming-Feng; Chang, Huan-Tsung
We applied surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) with HgTe nanostructures as the matrix for the detection of single- and double-stranded oligodeoxynucleotides (ss-ODNs and ds-ODNs). The concentrations of surfactant and additives (metal ions, an amine) and the pH and ionic strength of the sample matrix played significantly different roles in the detection of ss- and ds-ODNs with various sequences. In the presence of Brij 76 (1.5 %), Hg2+ (7.5 μM), and cadaverine (10 μM) at pH 5.0, this SALDI-MS approach allowed the simultaneous detection of T15, T20, T33, and T40, with limits of detection at the femtomole-to-picomole level and sample-to-sample intensity variation <23 %. In the presence of Ag+ (1 μM) and cadaverine (10 μM) at pH 7.0, this technique allowed the detection of randomly sequenced ss- and ds-ODNs at concentrations down to the femtomole level. To the best of our knowledge, this paper is the first to report the detection of ss-ODNs (up to 50-mer) and ds-ODNs (up to 30 base pairs) through the combination of SALDI-MS with HgTe nanostructures as matrices. We demonstrated the practicality of this approach through analysis of a single nucleotide polymorphism that determines the fate of the valine residue in the β-globin of sickle cell megaloblasts.
Taghavi Pourianazar, Negar; Gunduz, Ufuk
One major application of nanotechnology in cancer treatment involves designing nanoparticles to deliver drugs, oligonucleotides, and genes to cancer cells. Nanoparticles should be engineered so that they could target and destroy tumor cells with minimal damage to healthy tissues. This research aims to develop an appropriate and efficient nanocarrier, having the ability of interacting with and delivering CpG-oligodeoxynucleotides (CpG-ODNs) to tumor cells. CpG-ODNs activate Toll-like receptor 9 (TLR9), which can generate a signal cascade for cell death. In our study, we utilized three-layer magnetic nanoparticles composed of a Fe3O4 magnetic core, an aminosilane (APTS) interlayer and a cationic poly(amidoamine) (PAMAM) dendrimer. This will be a novel targeted delivery system to enhance the accumulation of CpG-ODN molecules in tumor cells. The validation of CpG-ODN binding to DcMNPs was performed using agarose gel electrophoresis, UV-spectrophotometer, XPS analyses. Cytotoxicity of conjugates was assessed in MDA-MB231 and SKBR3 cancer cells based on cell viability by XTT assay and flow cytometric analysis. Our results indicated that the synthesized DcMNPs having high positive charges on their surface could attach to CpG-ODN molecules via electrostatic means. These nanoparticles with the average sizes of 40±10nm bind to CpG-ODN molecules efficiently and induce cell death in MDA-MB231 and SKBR3 tumor cells and could be considered a suitable targeted delivery system for CpG-ODN in biomedical applications. The magnetic core of these nanoparticles represents a promising option for selective drug targeting as they can be concentrated and held in position by means of an external magnetic field. PMID:26898428
The decay of the radioisotope 125I into 125Te is typically followed by the emission of two groups of approximately 10 electrons each via Auger processes. In deoxyribonucleic acid (DNA) with 125I incorporated, these electrons produce various types of damage to DNA, e.g. single strand breaks (SSBs) and double strand breaks (DSBs) through direct actions of physical tracks, or indirect actions of radicals produced in water. Among the direct actions one should consider not only the excitation and ionisation of DNA by Auger electrons, but also the neutralisation of highly charged 125mTe ions with electrons from neighbouring molecules. Comparison between experiment and simulation done recently revealed that without including neutralisation effect the simulated yield of SSBs was 50% less than the measured result. In the present work a calculation of DNA strand breakage by the neutralization effect in a 41-mer synthetic oligodeoxynucleotide (oligo-DNA) model was done using the charge transfer theory. Calculation based on transfer rate using the newly evaluated electronic coupling of DNA bases showed that the positive charge (hole) transfer rate is of the order of magnitudes of several 1013s-1, implying that a charge higher than 10 units might not build on a 125mTe atom. The potential energy accumulated on the decay base is transferred to bases along the DNA chain nearby and destroys those bases and ionises the sugar-phosphate group, leading a DNA SSB with a frequency of 0.2% per eV in average. (authors)
张新霞; 崔长琮; 李江; 崔翰斌; 徐仓宝; 朱参战
Objective To investigate the effect of gelatin coated Platinium-Iridium stent absorbed c-myc antisense oligodeoxynucleotide (ASODN) on smooth muscle cells apoptosis in a normal rabbit carotid arteries. Methods Gelatin coated Platinium-Iridium stents were implanted in the right carotid arteries of 32 rabbits under vision. Animals were randomly divided into control group and treated group receiving c-myc ASODN (n=16, respectively). On 7, 14, 30 and 90 days following the stenting procedure ,morphometry for caculation of neointimal area and mean neointimal thickness were performed.The expression of c-myc protein was detected by immunohistochemical method. Apoptotic smooth muscle cells was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL). Results At 7 and 14 days after stenting,there were no detectable apoptotic cells in both groups. The apoptotic cells occurred in the neointima 30 and 90 days after stenting, and the number of apoptotic cells at 30 days were less [4.50±1.29 vs 25.75±1.89 (number/0.1mm2)] than that at 90 days [13.50±1.91 vs 41.50±6.46 (number/0.1mm2)]. Meanwhile c-myc ASODN induced more apoptotic cells than the control group(P<0.0001). c-myc protein expression was weak positive or negative in treated group and positive in control group.Conclusion c-myc ASODN can induce smooth muscle cells apoptosis after stenting in normal rabbit carotid arteries,and it can be used to prevent in-stent restenosis.
Hai-yan DU; Ai-guo JI; Zhong-ming TANG; Shao-you XIA; Hai-feng SONG; Na LI; Ming-mei SHANG; Jia ZOU; Qing-qing WANG; Lun OU; Xiao SUN
Aim: To study the relationship between primary structures of oligodeoxynucleo-tides (ODN) containing unmethylated deoxycytidyl-deoxyguanosine (CpG)dinucleotide motifs and their immunostimulatory activities in mouse spleen cells.Methods: A series of CpG ODN with different primary structures were synthesized.Their capabilities to stimulate mouse spleen cell proliferation were determined by[3H]thymidine incorporation assay. Cytokine (interleukin JILl-6, IL-12, and IFN-α)secretion spectra induced by CpG ODN were assessed by ELISA. The ability ofCpG ODN to activate natural killer cells was evaluated by standard 4 h 51Cr-release assay. Flow cytometry was utilized to examine the expressions of various lymphocyte surface molecules on diverse immunocytes. An effective CpG ODN for routine,ODN1826, was set as the template of modification and the positive control.Results: The immunostimulatory activities of CpG ODN with different sequences and compositions varied markedly, both in character and in extent. It was useless for improving the immunostimulatory activity of ODN1826 by simply increasing the functional hexameric CpG motif number, modifying the site of CpG motifs, or changing the distance between multi-CpG motifs. However, an addition of a selfcomplementary palindrome structure at the 3'-end, but not the 5'-end of CpG ODN,aroused marked improvement in its activity. Several designed ODN had superior comprehensive immunostimulatory properties compared to ODN 1826. Conclusion:The immunostimulatory activity ofa CpG ODN was relevant to its primary structure.It was useless for promoting immunostimulatory activity to simply change CpG motif number, space, or distance. The 3'-end palindrome structure of CpG ODN is associated with enhanced immunostimulatory activity.
Su, Hang; Yuan, Gailing; Su, Jianguo
CpG oligodeoxynucleotides (ODNs) show strong immune stimulatory activity in vertebrate, however, they possess specific sequence feature among species. In this study, we screened out an optimal CpG ODN sequence for grass carp (Ctenopharyngodon idella), 1670A 5'-TCGAACGTTTTAACGTTTTAACGTT-3', from six published sequences and three sequences designed by authors based on grass carp head kidney mononuclear cells and CIK (C. idella kidney) cells proliferation. VP4 mRNA expression was strongly inhibited by CpG ODN 1670A in CIK cells with GCRV infection, showing its strong antiviral activity. The mechanism via toll-like receptor 9 (TLR9)-mediated signaling pathway was measured by real-time quantitative RT-PCR, and TLR21 did not play a role in the immune response to CpG ODN. The late up-regulation of CiRIG-I mRNA expression indicated that RIG-I-like receptors (RLRs) signaling pathway participated in the immune response to CpG ODN which is the first report on the interaction between CpG and RLRs. We also found that the efficient CpG ODN can activates interferon system. Infected with GCRV, type I interferon expression was reduced and type II interferon was induced by the efficient CpG ODN in CIK cells, especially IFNγ2, suggesting that IFNγ2 played an important role in response to the efficient CpG ODN. These results provide a theoretical basis and new development trend for further research on CpG and the application of CpG vaccine adjuvant in grass carp disease control. PMID:26972738
Full Text Available Cytosine-guanine oligodeoxynucleotide (CpG ODN motifs of bacterial DNA are recognized through toll-like receptor 9 (TLR9 and are potent activators of innate immunity. However, the interaction between TLR9 and CpG ODN in aquatic species has not been well characterized. Hence, cobia TLR9 isoform B (RCTLR9B was cloned and its expression and induction in intestine were investigated. RCTLR9B cDNA consists of 3113bp encoding 1009 amino acids containing three regions, leucine rich repeats, transmembrane domain, and toll/interleukin-1 receptor (TIR domain. Intraperitoneal injection of CpG ODN 2395 upregulated RCTLR9 A and B and MyD88 and also induced the expressions of Mx, chemokine CC, and interleukin IL-1β. Cobia intraperitoneally injected with CpG ODN 1668 and 2395 had increased survival rates after challenge with Photobacterium damselae subsp. piscicida. In addition, formulation of CpG ODN with formalin-killed bacteria (FKB and aluminum hydroxide gel significantly increased expressions of RCTLR9 A (50 folds and B (30 folds isoforms at 10 dpi (CpG ODN 1668 and MyD88 (21 folds at 6 dpv (CpG ODN 2395. Subsequently, IL-1β increased at 6 dpv in 1668 group. No histopathological damage and inflammatory responses were observed in the injected cobia. Altogether, these results facilitate CpG ODNs as an adjuvant to increase bacterial disease resistance and efficacy of vaccines in cobia.
Full Text Available Abstract Background Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN are known to exert a strong adjuvant effect on Th1 immune responses. Although several genes have been reported, no comprehensive study of the gene expression profiles in human cells after stimulation with CpG ODN has been reported. Results This study was designed to identify a CpG-inducible gene cluster that potentially predicts for the molecular mechanisms of clinical efficacy of CpG ODN, by determining mRNA expression in human PBMC after stimulation with CpG ODN. PBMCs were obtained from the peripheral blood of healthy volunteers and cultured in the presence or absence of CpG ODN 2006 for up to 24 hours. The mRNA expression profile was evaluated using a high-density oligonucleotide probe array, GeneChip®. Using hierarchical clustering-analysis, out of a total of 10,000 genes we identified a cluster containing 77 genes as having been up-regulated by CpG ODN. This cluster was further divided into two sub-clusters by means of time-kinetics. (1 Inflammatory cytokines such as IL-6 and GM-CSF were up-regulated predominantly 3 to 6 hours after stimulation with CpG ODN, presumably through activation of a transcription factor, NF-κB. (2 Interferon (IFN-inducible anti-viral proteins, including IFIT1, OAS1 and Mx1, and Th1 chemoattractant IP-10, were up-regulated predominantly 6 to 24 hours after stimulation. Blocking with mAb against IFN-α/β receptor strongly inhibited the induction of these IFN-inducible genes by CpG ODN. Conclusion This study provides new information regarding the possible immunomodulatory effects of CpG ODN in vivo via an IFN-α/β receptor-mediated paracrine pathway.
YANG Ya-ying; YI yong-fen; ZHANG Xiao-yan; XIAO Chun-wei; LIN Xiao; ZHOU Wen-wen
Objective: To investigate the effect of MUC2 antisense oligodeoxynucleotide (ASODN) on cell proliferation, adhesion and proteolytic enzyme in human gastric carcinoma cell line (SGC7901). Methods: Phosphorothioate MUC2 ASODN was synthesized and packaged by lipofectin, and then transfected to SGC7901 cells. The expression of MUC2 mRNA and protein after transfection was detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical method respectively,and the effect of MUC2 ASODN on cell proliferation,adhesion and proteolytic enzyme was determined by flow cytometry(FCM), MTT method, Rose Bengal and immunohistochemical method. Results: Compared with the blank control group, ASODN efficiently downregulated the expression of MUC2 mRNA and protein in SGC7901 cells 48h after transfection(P＜0.01). Various concentrations of ASODN could significantly inhibit the growth of SGC7901, and the inhibition peaked at the 48th hour after transfection(P＜0.05). The apoptosis rate of the experimental group was about 4.38%, and the percentage of S-phase cells rose while G0/G1-phase cells fell because most of them were blocked at S-phase. In addition, cells treated with MUC2 ASODN showed lower adhesion ability with matrix and endothelial cells than control cells in vitro(P＜0.01). By immunohistochemical method, the upregulation of E-cadherin proteins and the downregulation of MMP2 and cathepsinD proteins were also observed(P＜0.05). Conclusion: MUC2 ASODN could efficiently inhibit SGC7901 cell proliferation, reduce cell adhesion ability and downregulate the expression levels of proteolytic enzyme in vitro.
This study was designed to determine serum Alpha-fetoprotein (AFP) and beta-h Chornionic Gonodotropin hormone (beta-hCG) levels among adult Pakistani population, and to observe their correlation. Serum AFP and beta-hCG levels were evaluated, using Micro-particle Enzyme Immuno Assay (MEIA) technology of M/s Abbott Laboratories. Data were compared using students t-test and correlation was computed. In the patients advised serum AFP assessment, 52% had normal AFP levels. comprising 37% male and 63% female subjects. For patients having a non-pathological picture, AFP levels varied non-significantly (p>0.05) between the Genders however, age varied highly significantly (p0.05) and highly significant (p0.05). Comparison of the population in the same age groups of both genders revealed significant (p0.05), due to a large standard error. Although the differences in beta-hCG levels were highly significant (p0.05). Among the studied cases, 67% patients had normal and 33% patients had raised beta-hCG levels. Beta-hCG levels show a decreasing trend with increasing age and beta-hCG levels were statistically significant (p<0.05) when patients under 50 years of age were compared with patients above 50 years. The coefficient of correlation between serum AFP and beta- hCG levels was 0.996454, which indicated a very strong. Significant positive correlation between the two tumour markers. The study showed that both serum AFP and beta-hCG are useful tumour markers and had a very strong positive correlation. (author)
Full Text Available Abstract Background To compare the outcome of IVF cycles in women receiving controlled ovarian stimulation with recFSH or recFSH plus low dose hCG. Methods A retrospective case control study, performed at a private practice affiliated with an academic institute. Patients were infertile women who were treated with IVF/ICSI and controlled ovarian stimulation in a long GnRH agonist protocol using either low dose hCG in addition to recFSH [N = 88] or recFSH alone [N = 99]. Primary outcomes were mean FSH dose, number of mature eggs, number of fertilized eggs, and serum levels of estradiol. Secondary outcomes were endometrial thickness, cycle cancellations and pregnancy rates. Results A significant increase in number of mature and fertilized eggs was observed in women over 40 years of age using low dose hCG in addition to recFSH. The estradiol level was significantly higher on the day of hCG administration and the serum level of FSH on cycle day 7 and on the day of hCG administration were lower. Conclusion Addition of low dose hCG to recFSH compared with recFSH alone significantly modified cycle characteristics in patients >/= 40 years and could be of potential benefit for IVF cycles in older infertile women.
Highlights: ► Amberlite CG-400 anion exchange resin has been used for the recovery of uranium. ► The breakthrough curves and elution behaviors of CG-400 resin have been studied in detailed. ► The mathematical models have been used to analyze the experimental data. ► The CG-400 resin has been applied successfully for uranium recovery from UCF liquid waste. - Abstract: Continuous fixed-bed column studies were carried out by using Amberlite CG-400 anion exchange resin for the recovery of uranium from aqueous solutions (synthetic solutions and uranium conversion facility (UCF) liquid waste). Effects of operating parameters such as flow rate and bed height were studied. The breakthrough capacity decreases with increasing flow rate, but this dependence was not significant with a long column. The maximum breakthrough capacity of uranium ions were achieved by CG-400 resin at a flow rate of 0.2 mL min−1 and bed height 9.1 cm (4 g resin). The elution behavior of uranium from CG-400 resin by various eluents have been investigated and the results show that 0.5 mol L−1 HNO3 is a good eluent for uranium recovery. The Adams–Bohart, Thomas, Yoon–Nelson and Dose–Response models were applied to the experimental data to determine the characteristic parameters of the column for process design using linear regression. The breakthrough curve calculated from the Dose–Response model was in best agreement with the experimental data
Full Text Available Abstract A simple, safe and cost-effective treatment protocol in ovarian stimulation is of great importance in IVF practice, especially in the case of previous unsuccessful attempts. hCG has been used as a substitute of LH because of the degree of homology between the two hormones. The main aim of this prospective randomized study was to determine, for the first time, whether low dose hCG added to rFSH for ovarian stimulation could produce better results compared to the addition of rLH in women entering IVF-ET, especially in those women that had previous IVF failures. An additional aim was to find an indicator that would allow us to follow-up ovarian stimulation and, possibly, modify it in order to achieve a better IVF outcome; and that indicator may be the cDNA copies of the LH/hCG receptor. Group A patients (n = 58 were administered hCG and Group B rLH (n = 56 in addition to rFSH in the first days of ovarian stimulation. The number of follicles and oocytes and, most importantly, implantation and pregnancy rates were shown to be statistically significantly higher in the hCG group. This study has also determined, for the first time to our best knowledge, m-RNA for LH/hCG receptors in the lymphocytes of peripheral blood 40 h before ovum pick-up. cDNA levels of the hCG receptor after ovarian stimulation were significantly higher among women receiving hCG compared to those receiving LH. In addition, higher levels were encountered among women with pregnancy compared to those without, although this was not statistically significant due to the small number of pregnancies. It seems that hCG permits a highly effective and more stable occupancy of rLH/hCG receptors and gives more follicles and more oocytes. The determination of cDNA copies could be, in the future, a marker during ovulation induction protocols and of course a predictor for the outcome of ART in the special subgroup of patients with previous failures.
王彦飞; 袁亚湘; 张洪超
In this paper we solve large scale ill-posed problems, particularly the image restoration problem in atmospheric imaging sciences, by a trust region-CG algorithm. Image restoration involves the removal or minimization of degradation (blur, clutter, noise, etc.) in an image using a priori knowledge about the degradation phenomena. Our basic technique is the so-called trust region method, while the subproblem is solved by the truncated conjugate gradient method, which has been well developed for well-posed problems.The trust region method, due to its robustness in global convergence, seems to be a promising way to deal with ill-posed problems.
The research aims to develop new and innovative learning and teaching methods, through the application of 3D CG technologies in Fashion and Textiles education. The research poster presents a series of investigations and experiments into the use of 3D technologies by BA/BSc & MA/MSc students. Case study methods are used to explore and gather data on practice and teaching and learning in HE fashion/textiles and clothing educational environments. 3D laser scanning technology, high cost and low...
张忠源; 胡望平; 池细俤; 叶桂云; 胡盈莹; 闫炳龙; 张永平
Objective To explore the rusults of human chorionicvilli gonadotropin (hCG) in serum (standard solution) with different detection system, different hospitals in Fujian Province and study their possible relationship with hCG relative molecular variants. Methods Beckman Access and Tosh AIA - 1800 immune machine detection system were used respectively to test the different concentrations of quality control things and the hCG of 44 patients. The hCG quality control data were collected from 2007 to 2009 in Fujian Province. And then literature to analysis the hCG molecular variants detection data were collected. Results Even with the same quality control things, the same serum and the same hCG relative molecular variants displayed different test results in different instrument detection system with a significant difference. The linear equation of Beckman ACCESS and Tosh AIA - 1800 was Y = 1. 3336 X - 70.199. The result of clinical serum hCG, quality control things in Tosh AIA - 1800 was 133. 8％ , 137. 2％ , and in Abbott was 132％ . These two results were higher than those of the Beckmen ACCESS which displayed 100％ . Conclusion The hCG molecular variants with different quality control things infulence the hCG detection results. Because of the hCG molecular variants, different instruments will display different hCG detection reuslts.%目的 探讨血清(标准物)中人绒毛膜促性腺激素(hCG) 在不同的检测系统、福建省各医院的测定结果,及其与hCG相关分子变异体可能的关系.方法 分别应用贝克曼Access、日本东曹(Tosh AIA-1800),在此两个免疫仪器检测系统检测不同浓度的质控物和44例患者血清的hCG;并收集福建省2007~2009年hCG质控数据、再收集文献hCG相关分子变异体检测数据进行分析.结果 同一质控物、同一血清、同一hCG相关分子变异体对不同的仪器检测系统,显示不同的测定结果,有显著差异.贝克曼Access与Tosh AIA-1800线性方程为Y=1
H. B. Kekre
Full Text Available This paper explores the novel idea for feature extraction based on bins approach for CBIR. This work has fulfilled all the criterion of efficient feature extraction method like dimensionality reduction, fast extraction and efficient retrieval. Main idea used in feature extraction is based on the histogram and the linear functions used to modify it. Each BMP image in database is separated in R, G and B planes. We have calculated the histogram for each of them which are modified using linear equations. These modified histograms are partitioned using CG so that mass of intensities of the image pixels will be distributed uniformly in two parts. This CG partitioning of three image planes leads to generate the eight bins. Information extracted from these bins is in the form of statistical first four absolute moments namely MEAN (MEAN, Standard deviation (STD, Skewness (SKEW, and Kurtosis (KURTO. Each of these moments are computed separately for R, G and B colors. This generates four types of feature vectors of dimension eight. Database of 2000 BMP images is used for the experimentation. Multiple feature vector databases are prepared as part of preprocessing work of this CBIR system. Comparison of query and database feature vector is carried out using three similarity measures namely Euclidean distance (ED, Absolute distance (AD and Cosine correlation distance (CD. To evaluate the retrieval efficiency of this system we have used three parameters, Precision Recall Cross over Point (PRCP, Longest String (LS and Length of String to Retrieve all Relevant (LSRR.
In this paper we present the coupling of MORSE-CG with the SLAC Unified Graphics System (UGS77) through an add-on package called MORSGRAF which allows for real-time display of neutron and photon tracks in the Monte Carlo simulation. In addition to displaying the myriad of complicated geometries that can be created with the MORSE Combinatorial Geometry program, MORSGRAF permits color tagging of neutrons (green) and photons (red) with the variation of track intensity an indicator of the energy of the particle. Particle types can be switched off and on by means of a mouse-icon system, and the perspective can be changed (i.e., rotated, translated, and zoomed). MORSGRAF also allows one to display the propagation of radiation through shields and mazes on an ordinary graphics terminal, as well as in documents printed on a laser printer. Several examples will be given to demonstrate the various capabilities of MORSGRAF coupled to MORSE-CG. 12 refs., 8 figs
Ratner, Laura D; Stevens, Guillermina; Bonaventura, Maria Marta; Lux-Lantos, Victoria A; Poutanen, Matti; Calandra, Ricardo S; Huhtaniemi, Ilpo T; Rulli, Susana B
The metabolic syndrome is a growing epidemic; it increases the risk for diabetes, cardiovascular disease, fatty liver, and several cancers. Several reports have indicated a link between hormonal imbalances and insulin resistance or obesity. Transgenic (TG) female mice overexpressing the human chorionic gonadotropin β-subunit (hCGβ+ mice) exhibit constitutively elevated levels of hCG, increased production of testosterone, progesterone and prolactin, and obesity. The objective of this study was to investigate the influence of hCG hypersecretion on possible alterations in the glucose and lipid metabolism of adult TG females. We evaluated fasting serum insulin, glucose, and triglyceride levels in adult hCGβ+ females and conducted intraperitoneal glucose and insulin tolerance tests at different ages. TG female mice showed hyperinsulinemia, hypertriglyceridemia, and dyslipidemia, as well as glucose intolerance and insulin resistance at 6 months of age. A 1-week treatment with the dopamine agonist cabergoline applied on 5-week-old hCGβ+ mice, which corrected hyperprolactinemia, hyperandrogenism, and hyperprogesteronemia, effectively prevented the metabolic alterations. These data indicate a key role of the hyperprolactinemia-induced gonadal dysfunction in the metabolic disturbances of hCGβ+ female mice. The findings prompt further studies on the involvement of gonadotropins and prolactin on metabolic disorders and might pave the way for the development of new therapeutic strategies. PMID:27154336
Full Text Available BACKGROUND: Antigen sparing and cross-protective immunity are regarded as crucial in pandemic influenza vaccine development. Both targets can be achieved by adjuvantation strategy to elicit a robust and broadened immune response. We assessed the immunogenicity of an inactivated H5N1 whole-virion vaccine (A/Vietnam/1194/2004 NIBRG-14, clade 1 formulated with emulsified nanoparticles and investigated whether it can induce cross-clade protecting immunity. METHODOLOGY/PRINCIPAL FINDINGS: After formulation with PELC, a proprietary water-in-oil-in-water nanoemulsion comprising of bioresorbable polymer/Span(R85/squalene, inactivated virus was intramuscularly administered to mice in either one-dose or two-dose schedule. We found that the antigen-specific serum antibody responses elicited after two doses of non-adjuvanted vaccine were lower than those observed after a single dose of adjuvanted vaccine, PELC and the conventional alum adjuvant as well. Moreover, 5 microg HA of PELC-formulated inactivated virus were capable of inducing higher antibodies than those obtained from alum-adjuvanted vaccine. In single-dose study, we found that encapsulating inactivated virus into emulsified PELC nanoparticles could induce better antibody responses than those formulated with PELC-adsorbed vaccine. However, the potency was rather reduced when the inactivated virus and CpG (an immunostimulatory oligodeoxynucleotide containing unmethylated cytosine-guanosine motifs were co-encapsulated within the emulsion. Finally, the mice who received PELC/CpG(adsorption-vaccine could easily and quickly reach 100% of seroprotection against a homologous virus strain and effective cross-protection against a heterologous virus strain (A/Whooper swan/Mongolia/244/2005, clade 2.2. CONCLUSIONS/SIGNIFICANCE: Encapsulating inactivated H5N1 influenza virus and CpG into emulsified nanoparticles critically influences the humoral responses against pandemic influenza. These results demonstrated
González Aguilera, B; Syrios, P; Gadisseur, R; Luyckx, F; Cavalier, E; Beckers, A; Valdes-Socin, H
Phantom hCG refers to persistent mild elevations of hCG, leading physicians to unnecessary treatments whereas neither a true hCG nor a trophoblastic disease is present. We report the case of a 23-year-old woman with persistent low levels of serum hCG detected one month after miscarriage. As choriocarcinoma was suspected, a chemotherapy trial of methotrexate was prescribed, without any hCG reduction. Subsequently, laparoscopy ruled out a trophoblastic residue and the patient was referred to the Endocrine Unit for further investigations. While low levels of hCG were still detected in serum, no hCG was detected in the urine. In addition, when serum was processed in a HBT tube for revealing heterophilic antibodies, hCG was no longer detected. Such finding indicated the presence of phantom hCG due to heterophilic mouse antibodies interaction. This case raises the need of clinico-biological discussion to avoid inappropriate therapeutic decisions. Based on this case experience and after review of the literature, we suggest that current gynecological protocols for the diagnosis and treatment of trophoblastic disease should consider the inclusion of urinary hCG and/or a test for serum heterophilic antibodies when appropriate. PMID:26792068
Full Text Available The aim this study was to compare two protocols of induction for ovulation by desloreline acetate and hCG in Quarter Horse mares. The choice of the animals was based on the observations by the estrus, by rectal palpation of the ovaries and by ultrassonography of the follicular dynamics. After estrus detection and follicle control, the measurement of the follicles and the classification of uterus were carried out. The animals that had dominant follicle (diameter more than 35 mm and swollen uterus were used. In these conditions, the mares received hCG or desloreline acetate. Once ovulation occurred, the artificial insemination was carried. Two groups were performed: G1 (20 animals received 1.5 mg desloreline acetate and G2 (20 animals received 1700 IU of hCG. Following 6h intervals, the control follicular was performed by ultrasonography. The follicular average diameter was 42.6 cm for the groups and set up a score of 0 to 3 of uterine edema displayed by the device as well as the time of ovulation. In conclusion, the desloreline acetate showed better performance than hCG, because the ovulation was induced in less time (nine hours than hCG (p<0.05.The pregnancy rate was 80 and 75 %, respectively in G1 and G2.
Abdel-Rehim, A M; Stathopoulos, Andreas; Orginos, Kostas
The technique that was used to build the EigCG algorithm for sparse symmetric linear systems is extended to the nonsymmetric case using the BiCG algorithm. We show that, similarly to the symmetric case, we can build an algorithm that is capable of computing a few smallest magnitude eigenvalues and their corresponding left and right eigenvectors of a nonsymmetric matrix using only a small window of the BiCG residuals while simultaneously solving a linear system with that matrix. For a system with multiple right-hand sides, we give an algorithm that computes incrementally more eigenvalues while solving the first few systems and then uses the computed eigenvectors to deflate BiCGStab for the remaining systems. Our experiments on various test problems, including Lattice QCD, show the remarkable ability of EigBiCG to compute spectral approximations with accuracy comparable to that of the unrestarted, nonsymmetric Lanczos. Furthermore, our incremental EigBiCG followed by appropriately restarted and deflated BiCGStab provides a competitive method for systems with multiple right-hand sides.
To elucidate the mechanism of oxaloacetate decarboxylation by Cg1458, recombinant Cg1458 has been purified and crystallized. Oxaloacetate decarboxylase catalyses the decarboxylation of oxaloacetate to pyruvate and CO2. Recently, the Corynebacterium glutamicum gene product Cg1458 was determined to be a soluble oxaloacetate decarboxylase. To elucidate the mechanism of oxaloacetate decarboxylation by Cg1458, recombinant Cg1458 was purified and crystallized. The best crystal was grown from 0.2 M MgCl2, 0.1 M Bis-Tris pH 6.0, 25%(w/v) polyethylene glycol 3350 using the hanging-drop method. The crystals belonged to space group P43212, with unit-cell parameters a = b = 124.1, c = 73.6 Å. The crystals are most likely to contain a dimer in the asymmetric unit, with a VM value of 2.27 Å3 Da−1. A full data set was collected at 1.9 Å resolution using synchrotron radiation on beamline BL17U of SSRF, Shanghai, China. Structure-solution attempts by molecular replacement were successful with PDB entries 3qdf or 2dfu as the template
Rodrigo Casquero Cunha
Full Text Available The cattle tick Rhipicephalus (Boophilus microplus is responsible for great economic losses. It is mainly controlled chemically, with limitations regarding development of resistance to the chemicals. Vaccines may help control this parasite, thereby reducing tick pesticide use. In this light, we performed subcloning of the gene of the protein Bm86-GC, the homologue protein that currently forms the basis of vaccines (GavacTM and TickGardPLUS that have been developed against cattle ticks. The subcloning was done in the pPIC9 expression vector, for transformation in the yeast Pichia pastoris. This protein was characterized by expression of the recombinant Mut+ strain, which expressed greater quantities of protein. The expressed protein (rBm86-CG was recognized in the Western-blot assay using anti-Gavac, anti-TickGard, anti-larval extract and anti-rBm86-CG polyclonal sera. The serum produced in cattle vaccinated with the antigen CG rBm86 presented high antibody titers and recognized the native protein. The rBm86-GC has potential relevance as an immunogen for vaccine formulation against cattle ticks.O carrapato-do-boi Rhipicephalus (Boophilus microplus é responsável por grandes perdas econômicas. Seu controle é principalmente químico e apresenta limitações quanto ao desenvolvimento de resistência aos princípios ativos. As vacinas podem auxiliar no controle deste parasita diminuindo as aplicações de carrapaticidas. Considerando isso, foi realizada a subclonagem do gene da proteína Bm86-CG, proteína homologa a que atualmente é a base das vacinas desenvolvidas (GavacTM e TickGardPLUS contra o carrapato-do-boi, no vetor de expressão pPIC9, para ser transformado em levedura, Pichia pastoris. Esta proteína foi caracterizada pela expressão da cepa recombinante Mut+ que expressou maior quantidade de proteína. A proteína expressa, rBm86-CG, foi reconhecida no ensaio de Western-blot pelos soros policlonais anti-Gavac, anti-TickGard, anti
Hari, Yoshiyuki; Matsugu, Sachiko; Inohara, Hiroyasu; Hatanaka, Yuri; Akabane, Masaaki; Imanishi, Takeshi; Obika, Satoshi
We succeeded in the synthesis of triplex-forming oligonucleotides (TFOs) that contain a deoxyribonucleotide (Py) bearing a 2-pyridine nucleobase or the 2',4'-BNA congener (Py(B)). By UV melting experiments, it was found that 2-pyridine was a very promising nucleobase for the sequence-selective recognition of a CG base pair within double-stranded DNA (dsDNA) in a parallel motif triplex. Moreover, Py(B) in TFOs showed stronger affinity to a CG base pair than Py with further increase in the selectivity. Using TFO including multiple Py(B) units, triplex formation with dsDNA containing three CG base pairs was observed. PMID:20648389
Full Text Available The evaluation of glycosyl composition is an essential step to guide future research designs applied in bioactivity. In the same way, the unexplored potential bioactivity of exopolysaccharide from Ganoderma lucidum is huge. Therefore, this study investigated the glycosyl composition of the exopolysaccharide isolated from submerged fermentation of G. lucidum to serve as guide for future studies on bioactivity. Glycosyl content and composition were evaluated by combined GC/MS of the TMS derivatives of the monosaccharide methyl glycosides produced from the sample by acidic methanolysis. Glycosyl composition analysis showed that the dominant carbohydrate component in all samples of exopolysaccharide isolated from submerged fermentation of G. lucidum CG 144 was glucose (58.1%, mannose (26.6% and galactose (12.5% which can be referred to as heteroglycan. These results suggest that this Ganoderma exopolysaccharide may be a new immunomodulatory agent.
Selvam, A M
The frequency distribution of DNA bases A, C, G, T exhibit fractal fluctuations, namely a zigzag pattern of an increase followed by a decrease of all orders of magnitude along the length of the DNA molecule. Selfsimilar fractal fluctuations are ubiquitous to space-time fluctuations of dynamical systems in nature. The power spectra of fractal fluctuations exhibit inverse power law form signifying long-range space-time correlations such that there is two-way communication between local (small-scale) and global (large-scale) perturbations. In this paper it is shown that DNA base CG frequency distribution in Takifugu rubripes (Puffer fish) Genome Release 4 exhibit universal inverse power law form of the statistical normal distribution consistent with a general systems theory model prediction of quantumlike chaos governing fractal space-time distributions. The model predictions are (i) quasicrystalline Penrose tiling pattern for the nested coiled structure thereby achieving maximum packing efficiency for the DNA m...
Selvam, A M
Power spectra of human DNA base C+G frequency distribution in all available contiguous sections exhibit the universal inverse power law form of the statistical normal distribution for the 24 chromosomes. Inverse power law form for power spectra of space-time fluctuations is generic to dynamical systems in nature and indicate long-range space-time correlations. A recently developed general systems theory predicts the observed non-local connections as intrinsic to quantumlike chaos governing space-time fluctuations of dynamical systems. The model predicts the following. (1) The quasiperiodic Penrose tiling pattern for the nested coiled structure of the DNA molecule in the chromosome resulting in maximum packing efficiency. (2) The DNA molecule functions as a unified whole fuzzy logic network with ordered two-way signal transmission between the coding and non-coding regions. Recent studies indicate influence of non-coding regions on functions of coding regions in the DNA molecule.
Full Text Available Abstract Background Matrix metalloproteases (MMPs constitute a family of enzymes capable of degrading different components of the extracellular matrix and are implicated in the invasion of tumor cells through the basement membrane. Polymorphisms in MMP genes may result in changes in the expression of MMPs being associated with the development and progression of cancer. We have investigated the association between three polymorphisms (-1607 1G/2G, +17 C/G and -77 A/G in the human collagenases MMP1, MMP8 and MMP13 and the risk of development or progression of lung cancer. Methods A hospital-based case-control study was designed including 501 lung cancer patients and 510 controls matched. Genotypes were determined by PCR-RFLP. Results were analyzed using unconditional logistic regression, Cox's proportional hazard regression, and the Kaplan-Meier method. Results The MMP1 and MMP13 promoter polymorphisms were not associated with lung cancer risk, while the C/G polymorphism in MMP8 was associated with a statistically significant decreased risk of developing lung cancer (ORadj = 0.65; 95%CI = 0.45–0.93. The Kaplan-Meier analysis showed that the polymorphisms in MMP1, MMP8 and MMP13 not seem to modify the overall survival. Multivariate analysis revealed that MMP1, MMP8 and MMP13 polymorphisms are not independent prognostic factors for overall survival. Conclusion This study suggests that the polymorphism in MMP8 is associated with a decreased lung cancer risk, which can be used as a prognostic marker in lung cancer.
Matrix metalloproteases (MMPs) constitute a family of enzymes capable of degrading different components of the extracellular matrix and are implicated in the invasion of tumor cells through the basement membrane. Polymorphisms in MMP genes may result in changes in the expression of MMPs being associated with the development and progression of cancer. We have investigated the association between three polymorphisms (-1607 1G/2G, +17 C/G and -77 A/G) in the human collagenases MMP1, MMP8 and MMP13 and the risk of development or progression of lung cancer. A hospital-based case-control study was designed including 501 lung cancer patients and 510 controls matched. Genotypes were determined by PCR-RFLP. Results were analyzed using unconditional logistic regression, Cox's proportional hazard regression, and the Kaplan-Meier method. The MMP1 and MMP13 promoter polymorphisms were not associated with lung cancer risk, while the C/G polymorphism in MMP8 was associated with a statistically significant decreased risk of developing lung cancer (ORadj = 0.65; 95%CI = 0.45–0.93). The Kaplan-Meier analysis showed that the polymorphisms in MMP1, MMP8 and MMP13 not seem to modify the overall survival. Multivariate analysis revealed that MMP1, MMP8 and MMP13 polymorphisms are not independent prognostic factors for overall survival. This study suggests that the polymorphism in MMP8 is associated with a decreased lung cancer risk, which can be used as a prognostic marker in lung cancer
Lo Piccolo, Luca; De Pasquale, Claudio; Fodale, Roberta; Puglia, Anna Maria; Quatrini, Paola
Enzymes involved in oxidation of long-chain n-alkanes are still not well known, especially those in Gram-positive bacteria. This work describes the alkane degradation system of the n-alkane degrader actinobacterium Gordonia sp. strain SoCg, which is able to grow on n-alkanes from dodecane (C12) to hexatriacontane (C36) as the sole C source. SoCg harbors in its chromosome a single alk locus carrying six open reading frames (ORFs), which shows 78 to 79% identity with the alkane hydroxylase (AH)...
Objective: To explore the effect of homeobox B2 (HOXB2) antisense oligodeoxynucleotides (asodn) on the proliferation and expression of primary human umbilical vein endothelial cells (HUVECs). Methods: Various concentrations of HOXB2 asodn modified by thiophosphate transfected the induction of liposome into HUVECs. MTT and RT-PCR methods were employed to determine the effect of different concentrations of asodn on the endothelial proliferation and the expression level of HOXB2 mRNA. Results: After the transfection of HOXB2 asodn, the endothelial proliferation was inhibited in a dose-dependent fashion. Simultaneously, the expression of HOXB2 mRNA decreased significantly. Conclusions: HOXB2 plays an important role in the proliferation of endothelia.
play key roles in autocrine mechanisms of bladder cancer cells remain to be definite and their physiological mechanism remain to be fully understood. More are still needed to know about the role of IGF1R signaling in bladder cancers. The following problems remain to be investigated in bladder cancer cells, about which the present studies are concerned: ①Is IGFs/IGF-IR signaling pathway involved in autocrine growth of human bladder cancer cells and how does bladder instillation drugs such as MMC affect the autocrine expression of bladder cancer cells? ②Can targeting against IGF1R gene can significantly enhance drug sensitivity of urinary bladder cancer cells to chemotherapy? ③What potential intracellular signaling mechanisms are involved in IGF1R blockage? ④May IGF1R self-stablized antisense ODN serves as a potential therapeutic approach to bladder cancer? To investigate whether IGF-1R was involved in drug resistance of bladder cancer cells. METHODS: RT-PCR was used to detect the mRNA expression of IGF-I, IGF-Ⅱ, and IGF-IR in T24 cells and normal urothelial cells. Flow cytometry and MTT tests were used to assess the effect of antisense oligodeoxynucleotide (ODN) on drug sensitivities and apoptosis of T24 cells to mitomycin (MMC). Western blot was used to analyze the effect of ODN on expression of IGF-IR protein. RESULTS: mRNA of IGF-I, IGF-Ⅱ, and IGF-IR were strongly expressed in serum-free cultured T24 cell line, whereas normal urothelial cells did not express these factors/receptors or only in trace levels; knockdown of IGF1R by antisense ODN significantly inhibited the growth of bladder cancer cells and enhanced sensitivity and apoptosis of T24 cells to MMC. CONCLUSION: These results suggested that blockage of IGF-IR signaling might potentially contribute to the treatment of bladder cancer cells which are insensitive to chemotherapy.
于波; 辛毅; 赵春江; 吴常信; 邓学梅
In this study,the CG42741 gene expression were found at embryonic,larval and pupal stage by RT-PCR,but such expression was very low at all of the three stages.Comparative to embryonic and larval stages,the expression of the gene was higher at pupal stage.The structural analysis and functional prediction of CG42741 protein also conducted in Drosophila melanogaster.Our data showed that this protein was characterized by three Cys2His2 zinc fingers located at the C-terminus and executed its function in nucleus.The protein belonged to the Kruppel-like family of transcription factors and could bind with GC-rich sites in genomic DNA.And we inferred the CG42741 protein might play an important role in cell proliferation.%为深入了解黑腹果蝇CG42741基因的表达及其编码蛋白质的结构,利用半定量和生物信息学的方法对其进行了详细的研究。RT-PCR结果显示黒腹果蝇CG42741基因在胚胎期、幼虫期和蛹期均有表达,但表达量比较低,只是在蛹期表达量相对升高。此外,通过生物信息学的方法对果蝇中CG42741基因所编码的蛋白质进行了结构与功能的预测。结果显示,CG42741蛋白定位于细胞核内,其羧基末端含有3个串联的C2H2类型的锌指结构;它属于KLF家族蛋白,是一个转录调控因子,能够与富含GC的基因组序列特异结合,其功能可能与细胞
Full Text Available Abstract Background Both apolipoprotein (Apo C-III gene polymorphism and alcohol consumption have been associated with increased serum triglyceride (TG levels, but their interactions on serum TG levels are not well known. The present study was undertaken to detect the interactions of the ApoC-III 3238C>G (rs5128 polymorphism and alcohol consumption on serum TG levels. Methods A total of 516 unrelated nondrinkers and 514 drinkers aged 15-89 were randomly selected from our previous stratified randomized cluster samples. Genotyping of the ApoC-III 3238C>G was performed by polymerase chain reaction and restriction fragment length polymorphism combined with gel electrophoresis, and then confirmed by direct sequencing. Interactions of the ApoC-III 3238C>G genotype and alcohol consumption was assessed by using a cross-product term between genotypes and the aforementioned factor. Results Serum total cholesterol (TC, TG, high-density lipoprotein cholesterol (HDL-C, ApoA-I and ApoB levels were higher in drinkers than in nondrinkers (P P P P P P P P Conclusions This study suggests that the ApoC-III 3238CG heterozygotes benefited more from alcohol consumption than CC and GG homozygotes in increasing serum levels of HDL-C, ApoA-I, and the ratio of ApoA-I to ApoB, and lowering serum levels of TC and TG.
Full Text Available Corynebacteria contain a heme uptake system encoded in hmuTUV genes, in which HmuT protein acts as a heme binding protein to transport heme to the cognate transporter HmuUV. The crystal structure of HmuT from Corynebacterium glutamicum (CgHmuT reveals that heme is accommodated in the central cleft with His141 and Tyr240 as the axial ligands and that Tyr240 forms a hydrogen bond with Arg242. In this work, the crystal structures of H141A, Y240A, and R242A mutants were determined to understand the role of these residues for the heme binding of CgHmuT. Overall and heme environmental structures of these mutants were similar to those of the wild type, suggesting that there is little conformational change in the heme-binding cleft during heme transport reaction with binding and the dissociation of heme. A loss of one axial ligand or the hydrogen bonding interaction with Tyr240 resulted in an increase in the redox potential of the heme for CgHmuT to be reduced by dithionite, though the wild type was not reduced under physiological conditions. These results suggest that the heme environmental structure stabilizes the ferric heme binding in CgHmuT, which will be responsible for efficient heme uptake under aerobic conditions where Corynebacteria grow.
Al Humaidan, Peter Samir Heskjær; Polyzos, N P; Alsbjerg, B;
risk of OHSS the administration of two boluses of 1.500 IU hCG after GnRHa trigger should be avoided as it may induce OHSS. STUDY FUNDING/POTENTIAL COMPETING INTERESTS: The study was supported by a research grant by MSD, Denmark. No conflict of interest was declared. TRIAL REGISTRATION NUMBER: Project...
Brouwers, AH; Buijs, WCAM; Mulders, PFA; de Mulder, PHM; van den Broek, WJM; Mala, C; Oosterwijk, E; Boerman, OC; Corstens, FHM; Oyen, WJG
Purpose: A study was designed to define the therapeutic efficacy, safety, and toxicity of two sequential high-dose treatments of radioimmunotherapy with [I-131]cG250 in patients with metastasized renal cell carcinoma. Here, we report the dosimetric analysis and the relationship between the developme
Gent, 2001. s. 7. [Contextualizing the Genome : the Role of Epigenetics in Genetics, Development and Evolution. 25.11.2001-28.11.2001, Gent] Institutional research plan: CEZ:AV0Z5004920 Keywords : CG and CNG hypomethylation * epigenetic stability * tobacco genome Subject RIV: BO - Biophysics
Nicalon-CG and Hi-Nicalon fibers were characterized by measuring their length, density, and tensile strength in the unirradiated, thermal annealed, and irradiated conditions. The irradiation was conducted in the EBR-II to a dose of 43 dpa-SiC at a nominal irradiation temperature of 1,000 C. The annealed specimens were held at 1,010 C for 165 days to approximately duplicate the thermal exposure of the irradiated specimens. The results indicate the fibers that perform best in an irradiation environment are those that approach stoichiometric and crystalline SiC. Hi-Nicalon exhibited negligible densification, accompanied by an increase in tensile strength after irradiation. Nicalon-CG possessed a higher tensile strength than hi-Nicalon in the unirradiated condition, but was significantly weakened in the annealed and irradiated conditions. In addition, Nicalon-CG exhibited unacceptable irradiation-induced shrinkage. Loss o fiber tensile strength after irradiation is shown to reduce the flexural strength of irradiated composites and Nicalon-CG fiber shrinkage observed in irradiated composites
Thuesen, L L; Andersen, A Nyboe; Loft, A;
GnRH agonist protocol with recombinant FSH 150 IU/d and randomized from stimulation day 1 to supplementation with hCG: D0, 0 IU/d; D50, 50 IU/d; D100, 100 IU/d; and D150, 150 IU/d. MAIN OUTCOME MEASURE: Intrafollicular hormone concentrations in relation to treatment groups, follicular sizes, and...
Muraki, Norifumi; Kitatsuji, Chihiro; Ogura, Mariko; Uchida, Takeshi; Ishimori, Koichiro; Aono, Shigetoshi
Corynebacteria contain a heme uptake system encoded in hmuTUV genes, in which HmuT protein acts as a heme binding protein to transport heme to the cognate transporter HmuUV. The crystal structure of HmuT from Corynebacterium glutamicum (CgHmuT) reveals that heme is accommodated in the central cleft with His141 and Tyr240 as the axial ligands and that Tyr240 forms a hydrogen bond with Arg242. In this work, the crystal structures of H141A, Y240A, and R242A mutants were determined to understand the role of these residues for the heme binding of CgHmuT. Overall and heme environmental structures of these mutants were similar to those of the wild type, suggesting that there is little conformational change in the heme-binding cleft during heme transport reaction with binding and the dissociation of heme. A loss of one axial ligand or the hydrogen bonding interaction with Tyr240 resulted in an increase in the redox potential of the heme for CgHmuT to be reduced by dithionite, though the wild type was not reduced under physiological conditions. These results suggest that the heme environmental structure stabilizes the ferric heme binding in CgHmuT, which will be responsible for efficient heme uptake under aerobic conditions where Corynebacteria grow. PMID:27240352
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Spencer, K; Tul, N; Nicolaides, K H
We have studied maternal serum free beta-hCG and PAPP-A, and fetal nuchal translucency (NT) in a series of 46 cases of fetal Turner's syndrome, 13 cases of other sex chromosomal anomalies and compared these with 947 control pregnancies in the first trimester. In cases of Turner's syndrome (45,X) the median fetal NT was significantly higher than in controls (4.76 MoM), the median PAPP-A was significantly lower (0.49 MoM), whilst the free beta-hCG was not significantly different (1.11 MoM). For NT, 93% (43/46) of cases were equal to or greater than the 95th centile of controls, for PAPP-A 35% (16/46) of cases were less than or equal to the 5th centile of controls and for free beta-hCG 15% (7/46) of cases were equal to or greater than the 95th centile of controls. For other sex chromosomal anomalies (47XXX, XXY, XYY) the median NT was increased (2.07 MoM) whilst PAPP-A was not significantly decreased (0.88 MoM) and free beta-hCG was not significantly different (1.07 MoM) from controls. Using a previously derived multivariate risk algorithm for trisomy 21, incorporating NT, PAPP-A, free beta-hCG and maternal age, 96% of the Turner's cases and 62% of the other sex chromosomal anomalies would have been identified. PMID:10820406
Liu, Xin, E-mail: email@example.com [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China); Zhu, Yanming, E-mail: firstname.lastname@example.org [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China); Zhai, Hong, E-mail: Zhai.email@example.com [Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Harbin 150040 (China); Cai, Hua, E-mail: firstname.lastname@example.org [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China); Ji, Wei, E-mail: email@example.com [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China); Luo, Xiao, E-mail: firstname.lastname@example.org [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China); Li, Jing, E-mail: email@example.com [Plant Secondary Metabolism Laboratory, Northeast Agricultural University, Harbin 150030 (China); Bai, Xi, E-mail: firstname.lastname@example.org [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China)
Highlights: Black-Right-Pointing-Pointer AtPP2CG1 positively regulates salt tolerance in ABA-dependent manner. Black-Right-Pointing-Pointer AtPP2CG1 up-regulates the expression of marker genes in different pathways. Black-Right-Pointing-Pointer AtPP2CG1 expresses in the vascular system and trichomes of Arabidopsis. -- Abstract: AtPP2CG1 (Arabidopsis thaliana protein phosphatase 2C G Group 1) was predicted as an abiotic stress candidate gene by bioinformatic analysis in our previous study. The gene encodes a putative protein phosphatase 2C that belongs to Group G of PP2C. There is no report of Group G genes involved in abiotic stress so far. Real-time RT-PCR analysis showed that AtPP2CG1 expression was induced by salt, drought, and abscisic acid (ABA) treatment. The expression levels of AtPP2CG1 in the ABA synthesis-deficient mutant abi2-3 were much lower than that in WT plants under salt stress suggesting that the expression of AtPP2CG1 acts in an ABA-dependent manner. Over-expression of AtPP2CG1 led to enhanced salt tolerance, whereas its loss of function caused decreased salt tolerance. These results indicate that AtPP2CG1 positively regulates salt stress in an ABA-dependent manner. Under salt treatment, AtPP2CG1 up-regulated the expression levels of stress-responsive genes, including RD29A, RD29B, DREB2A and KIN1. GUS activity was detected in roots, leaves, stems, flower, and trichomes of AtPP2CG1 promoter-GUS transgenic plants. AtPP2CG1 protein was localized in nucleus and cytoplasm via AtPP2CG1:eGFP and YFP:AtPP2CG1 fusion approaches.
To observe the liver condition of hyperthyroid patients with different course of disease, the levels of serum HA, C-IV and CG in 124 hyperthyroid patients and 60 normal persons were measured with RIA. Hyperthyroid patients were divided into 3 groups by their different course of disease and the results were managed statistically. For normal persons and the group whose course of disease were within half year, the levels of all 3 objects are in normal range. For the group whose course of disease were in period of 0.5-2 years, the levels of HA and C-IV are over the normal range. For the group whose course of disease were over 2 years, the levels of all 3 objects are over the normal range and the levels of HA and C-IV are much higher than the second group. The results suggested that hyperthyroid patients whose course of disease are over half year would suffer from hepatocellular damage and fibrosis and the damage degree is parallel to their courses of disease
"Cosmologists are often in error but never in doubt." This pithy characterization by the Soviet physicist Lev Landau sums up the raison d'être of Facts and Speculations in Cosmology. Authors Jayant Narlikar and Geoffrey Burbidge are proponents of a "steady state" theory of cosmology, and they argue that the cosmological community has become fixated on a "Big Bang" dogma, suppressing alternative viewpoints. This book very much does what it says on the tin: it sets out what is known in cosmology, and puts forward the authors' point of view on an alternative to the Big Bang.
Bagger-Petersen, Mai Corlin
From 2014, Anhui Province will pilot a reform of the residential land market in China, thus integrating rural Anhui in the national housing market. In contrast, artist and activist Ou Ning has proposed the Bishan time money currency, intending to establish an alternative economic circuit in Bishan...
Hoette, Vivian L.; Rebull, L. M.; McCarron, K.; Johnson, C. H.; Gartner, C.; VanDerMolen, J.; Gamble, L.; Matche, L.; McCartney, A.; Doering, M.; Crump, R.; Laorr, A.; Mork, K.; Steinbergs, E.; Wigley, E.; Caruso, S.; Killingstad, N.; McCanna, T.
Individuals with disabilities - specifically individuals who are deaf or hard of hearing (DHH) and/or blind and visually-impaired (BVI) - have traditionally been underrepresented in the fields of Science, Technology, Engineering, and Math (STEM). The low incidence rate of these populations, coupled with geographic isolation, creates limited opportunities for students to work with and receive mentoring by professionals who not only have specialty knowledge in disability areas but also work in STEM fields. Yerkes Observatory scientists, along with educators from the Wisconsin School for the Deaf, the Wisconsin Center for the Blind and Visually Impaired, Breck School, and Oak Park and River Forest High School, are engaged in active research with a Spitzer Science Center (SSC) scientist. Our ultimate goals are threefold; to engage DHH and BVI students with equal success as their sighted and hearing peers, to share our techniques to make astronomy more accessible to DHH and BVI youth, and to generate a life-long interest which will lead our students to STEM careers. This poster tracks our work with an SSC scientist during the spring, summer, and fall of 2010. The group coauthored another AAS poster on finding Young Stellar Objects (YSO) in the CG4 Nebula in Puppis. During the project, the students, scientists and teachers developed a number of techniques for learning the necessary science as well as doing the required data acquisition and analysis. Collaborations were formed between students with disabilities and their non-disabled peers to create multi-media projects. Ultimately, the projects created for our work with NITARP will be disseminated through our professional connections in order to ignite a passion for astronomy in all students - with and without disabilities. This research was made possible through the NASA/IPAC Teacher Archive Research Project (NITARP) and was funded by NASA Astrophysics Data Program and Archive Outreach funds.
Pharmacokinetic Profile and Acute Toxicological Properties of a Novel Radiosensitizer Cytosine-Phosphate-Guanosine Oligodeoxynucleotide 107 in Mice Following Intravenous and Orthotopic Administration.
Cen, Yanyan; Li, Xiaoli; Yin, Zhiwei; Yan, Zifei; Liu, Dan; Peng, Wei; Pan, Feng; Zhou, Hong
The synthetic cytosine-phosphate-guanosine oligodeoxynucleotide 107 (CpG ODN107) is a novel radiosensitizer for glioma treatment. However, the information related to its pharmacokinetics and toxicity remains unclear. Therefore, the plasma pharmacokinetics, distribution, elimination, and acute toxicity of CpG ODN107 in mice were investigated in the present experiments. The results from the liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay showed that the plasma elimination half-life (t1/2β) of CpG ODN107 in BALB/c mice varied slightly with the dose, and it was 0.65, 0.49, and 0.50 h at the intravenous doses of 2.5, 5, and 10 mg/kg, respectively. CpG ODN107 rapidly and widely distributed in organs/tissues, except the brain and testes. The highest concentrations were found in the liver (28.6% of the administered dose after 0.5 h) and the kidneys (5.7% of the administered dose after 1 h). CpG ODN107 (0.3, 3, and 30 μg/mL) could highly bind to human and mouse plasma proteins in vitro. CpG ODN107 in the forms of prototype was excreted in urine (1.79%) and feces (0.91%), and its shortened metabolites were excreted in urine (2.1%) and feces (2.2%) within the first 24 h. The mice in vivo optical image showed CpG ODN107 labeled with Alexa Fluor 680 fluorochrome (AF680) accumulated in the brain after orthotopic injection, eliminated very slowly, and excreted in urine compared with poly T labeled with AF680. The median lethal dose (LD50) of CpG ODN107 was 75.7 mg/kg for mice; this dose only could produce apparent spleen and liver damage, in line with the distribution features of CpG ODN. In conclusion, our present pharmacokinetic and toxicity investigation will provide helpful information to further pharmacodynamic and pharmacokinetic research of CpG ODN107 and other oligodeoxynucleotide drugs in the future. PMID:26213852
Full Text Available Liu Huining,1 Zhang Yi,1 Tang Dihong,2 Pan Yifeng,3 Xia Man,2 Yang Ting,2 Cai Jingting1,2 1Department of Obstetrics and Gynecology, Xiangya Hospital, 2Department of Gynecological Oncology, Hunan Provincial Tumor Hospital, 3National Hepatobiliary and Enteric Surgery Research Center, Central South University, Changsha, Hunan, People's Republic of China Objective: To observe the influence of Fe3O4-dextran-anti-ß-human chorionic gonadotropin (HCG carrying heparanase (Hpa antisense oligodeoxynucleotide (ASODN, via the invasion, proliferation, and Hpa expression of JEG-3 cell lines and inhibitory effect of transplanted choriocarcinoma tumor growth. Methods: The different abilities of invasion and proliferation between transfected JEG-3 and untransfected JEG-3 were measured by Matrigel invasion assay and 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide assay in vitro. The effect of Hpa ASODN transfection on the expression of Hpa mRNA and protein was measured by reverse-transcription polymerase chain reaction and Western blot. The transplanted choriocarcinoma tumors were taken out to calculate the inhibitory effect on tumor growth of Hpa ASODN. Results: In this study, we found that: (1 the invasive ability of JEG-3 cells was inhibited sufficiently (P < 0.05 after JEG-3 cells were transfected by Fe3O4-dextran-anti-βHCG carrying Hpa ASODN; (2 after JEG-3 cells were transfected by Fe3O4-dextran-anti-βHCG carrying Hpa ASODN at 48 and 72 hours, the proliferative ability of JEG-3 cells was inhibited sufficiently (P < 0.05; (3 the expression of Hpa mRNA and protein in JEG-3 cells was inhibited efficiently after JEG-3 cells were transfected by Fe3O4-dextran-anti-βHCG carrying Hpa ASODN (P < 0.05; and (4 Fe3O4-dextran-anti-βHCG carrying Hpa ASODN had an inhibitory effect on the transplanted choriocarcinoma tumor growth (P < 0.05 and was harmless on nude mice. Conclusion: Fe3O4-dextran-anti-βHCG carrying Hpa ASODN weakened the invasive and
Sato, Tomoaki; Nagaoka, Keizo; Oguchi, Kosei
A CG-animation content which supports the learning of the Otto cycle was developed. This content has a piston assembly and the diagrams of PV, VS, TP and TS. The each diagram has a pointer which moves along the line of the graph and they are synchronized with the movement of the piston. The learners can operate this content directly on the e-learning system. While watching the movements of the piston assembly, the learners can confirm the state of the engine about temperature, pressure, volume, and entropy by the synchronized pointer on the diagrams. This content was used for the class of the machining practice exercise. The learning effect of the content was examined by the score of the short test. As the result of this examination, the CG-animation content was effective in the learning of the Otto cycle.
Bogdanovich, Tatiana; Clark, Catherine; Kosowska-Shick, Klaudia; Dewasse, Bonifacio; McGhee, Pamela; Appelbaum, Peter C.
Among 203 strains of Staphylococcus aureus, the MICs of CG400549 were 0.06 to 1.0 μg/ml, with MIC50 and MIC90 values of 0.25 μg/ml each. All strains were susceptible to linezolid and quinupristin-dalfopristin (MICs, 0.25 to 2.0 μg/ml). The daptomycin MICs were 0.25 to 2.0 μg/ml for methicillin-susceptible and 0.25 to 4.0 μg/ml against methicillin-resistant strains (including vancomycin-intermediate strains). Single-passage selection testing showed low resistance frequencies with CG400549, but...
Risselada, Arne J; Vehof, Jelle; Bruggeman, Richard; Wilffert, Bob; Cohen, Dan; Al Hadithy, Asmar F; Arends, Johan; Mulder, Hans
The prevalence of the metabolic syndrome is increased in patients with schizophrenia compared with the general population. The strong interindividual differences in susceptibility to developing the metabolic syndrome suggests that the genetic makeup is a modulating factor. Part of the genetic puzzle can possibly be explained by variations in the gene coding for the adrenergic α-2a receptor (ADRA2A) because this receptor plays an important role in lipolysis. Three studies have found an association between the α-2a 1291-C/G polymorphism and antipsychotic induced weight gain, with conflicting results between whites and Asians. No studies have been published investigating the association between the 1291-C/G polymorphism and the metabolic syndrome. The primary objective of this cross-sectional study was to investigate the association between the ADRA2A 1291-C/G polymorphism and the metabolic syndrome in 470 patients using antipsychotic drugs. There was no significant association between carriership of the variant 1291-G allele and prevalence of the metabolic syndrome (odds ratio, 0.73; 95% confidence interval, 0.49-1.15). Exploratory analysis showed an association between carriership of the variant 1291-G allele and a reduced prevalence of the metabolic syndrome in patients not currently using antipsychotics (odds ratio, 0.05; 95% confidence interval, 0.003-0.97; P = 0.048). In conclusion, this study shows that the ADRA2A 1291-C/G polymorphism does not seem to be a strong predictor for long-term occurrence of the metabolic syndrome in antipsychotic using patients. Studies investigating this association using a prospective, or retrospective, design, as well as studies investigating this association in a nonpsychiatric population, are warranted. PMID:21105277
Tatiana Figueiredo; Roman Paiva; Luiz Ernandes Kozicki; Fernanda Kaercher; Romildo Romualdo Weiss; Ivo Walter dos Santos; Priscilla Ricabone Muradas
The aim this study was to compare two protocols of induction for ovulation by desloreline acetate and hCG in Quarter Horse mares. The choice of the animals was based on the observations by the estrus, by rectal palpation of the ovaries and by ultrassonography of the follicular dynamics. After estrus detection and follicle control, the measurement of the follicles and the classification of uterus were carried out. The animals that had dominant follicle (diameter more than 35 mm) and swollen ut...
Ratner, Laura D.; Gonzalez, Betina; Ahtiainen, Petteri; Di Giorgio, Noelia P.; Poutanen, Matti; Calandra, Ricardo S.; Ilpo T Huhtaniemi; Rulli, Susana B.
Female infertility is often associated with deregulation of hormonal networks, and hyperprolactinemia is one of the most common endocrine disorders of the hypothalamic-pituitary axis affecting the reproductive functions. We have shown previously that transgenic female mice overexpressing human chorionic gonadotropin β-subunit (hCGβ+ mice), and producing elevated levels of bioactive LH/hCG, exhibit increased production of testosterone and progesterone, are overweight and infertile, and develop...
Session Donna R
Full Text Available Abstract Background Administration of recombinant luteinizing hormone (rLH in controlled ovarian hyperstimulation may benefit a subpopulation of patients. However, late follicular phase administration of high doses of rLH may also reduce the size of the follicular cohort and promote monofollicular development. Methods To determine if rLH in late follicular development had a negative impact on follicular growth and oocyte yield, IVF patients in our practice who received rFSH and rLH for the entire stimulation were retrospectively compared with those that had the rLH discontinued at least two days prior to hCG trigger. Results The two groups had similar baseline characteristics before stimulation with respect to age, FSH level and antral follicle count. However, the group which had the rLH discontinued at least two days prior to their hCG shot, had a significantly higher number of oocytes retrieved, including a higher number of MII oocytes and number of 2PN embryos. Conclusions When using rLH for controlled ovarian hyperstimulation, administering it from the start of stimulation and stopping it in the late follicular phase, at least two days prior to hCG trigger, may increase oocyte and embryo yield.