Detection of siRNA administered to cells and animals by using a fluorescence intensity distribution analysis polarization system

Small interfering RNA (siRNA) has excellent pharmacological features and is expected to be used for therapeutic drug development. To this end, however, new RNA technology needs to be established so that extremely small amounts (less than 1 pmol) of siRNA can be detected in organs of experimental animals and in human blood to facilitate pharmacokinetics studies. An important feature is that this new technology is not dependent on radioisotopes and can detect siRNA molecules identical to those used for drug development in preclinical tests with experimental animals or in clinical tests with humans. We report a convenient method that can detect small amounts of siRNA. The method uses high-power confocal microscopic analysis of fluorescence polarization in DNA probes that are bound to one of the strands of siRNA and directly quantitates the copy number of siRNA molecule after extraction from specimens. A pharmacokinetic study to examine the blood retention time of siRNA/cationic liposomes in mice showed that this straightforward method is consistent with the other reverse transcriptase polymerase chain reaction amplification-based method. We believe that the entire process is simple and applicable for a high-throughput analysis, which provides excellent technical support for fundamental research on RNA interference and development of siRNA drugs

Praktična sinteza regulatora za precizno pozicioniranje sustava pomične podloge

This paper presents a practical feedback controller design of a ball screw-driven table system for the microdisplacement positioning. Friction of the mechanism in the micro-displacement region has nonlinear elastic properties, unlike Coulomb and/or viscous friction in the macro-displacement, resulting in different positioning responses and frequency characteristics of the plant depending on the regions. In this paper, at first, a numerical simulator with a rolling friction model is adopted to reproduce the positioning behaviors in the micro-displacement region. Based on the simulator, the stability condition of positioning in the region is clarified on the basis of frequency characteristics and, then, appropriate parameters of feedback controller are practically designed to satisfy the required positioning performance. Effectiveness of the proposed design has been verified by a series of experiments using a prototype of ball screw-driven table positioning device.U radu je prikazana sinteza regulatora s povratnom vezom u sustavu za precizno linearno pozicioniranje pomične podloge pomoću kugličnih ležajeva. Za razliku od uobičajenih modela Coulombova i/ili viskoznog trenja, trenje razmatranog sustava ima izrazito nelinearna svojstva u području mikro-pomaka, što za posljedicu ima različite odzive pozicioniranja i frekvencijski karakteristike, ovisno o radnom području. U radu je prvo razvijeno numeričko simulacijsko okruženje zasnovano na modelu trenja kotrljanja u svrhu simuliranja ponašanja sustava pozicioniranja u području mikropomaka. Potom je, zasnivajući se na simulacijskom okruženju, pomoću frekvencijske karakteristike razjašnjen problem stabilnosti sustava u promatranom radnom području te su odabrani odgovarajući parametri regulatora koji poštuju uvjet stabilnosti i zadovoljavaju željenu kvalitetu odziva. Sinteza regulatora provedena je vodeći računa o praktičnoj primjenjivosti postupka. Učinkovitost predložene sinteze potvr.ena je nizom eksperimenata na prototipu sustava za precizno linearno pozicioniranje pomične podloge pomoću kugličnih ležajeva

Nonrigid chiral soliton for the octet and decuplet baryons

Systematic treatment of the collective rotation of the nonrigid chiral soliton is developed in the SU(3) chiral quark soliton model and applied to the octet and decuplet baryons. The strangeness degrees of freedom are treated by a simplified bound-state approach which omits the locality of the kaon wave function. Then, the flavor rotation is divided into the isospin rotation and the emission and absorption of the kaon. The kaon Hamiltonian is diagonalized by the Hartree approximation. The soliton changes the shape according to the strangeness. The baryons appear as the rotational bands of the combined system of the soliton and the kaon.Comment: 11 pages(LaTex), 1 figures(eps

Instability of the hedgehog shape for the octet baryon in the chiral quark soliton model

In this paper the stability of the hedgehog shape of the chiral soliton is studied for the octet baryon with the SU(3) chiral quark soliton model. The strangeness degrees of freedom are treated by a simplified bound-state approach, which omits the locality of the kaon wave function. The mean field approximation for the flavor rotation is applied to the model. The classical soliton changes shape according to the strangeness. The baryon appears as a rotational band of the combined system of the deformed soliton and the kaon.Comment: 24 pages, LaTeX, 8 eps file

Partially conserved axial current constraints on pion production/absorption within nonrelativistic dynamics

We show the necessity of two-nucleon axial currents and associated pion emission/ absorption operators for the partial conservation of the axial current (PCAC) nuclear matrix elements with arbitrary nuclear dynamics described by a nonrelativistic Schroedinger equation. As examples we construct such nonrelativistic axial two-body currents in the linear- and heterotic (g_A = 1.26) sigma models, with an optional isoscalar vector (omega) meson exchange. The nuclear matrix elements obey PCAC only if the nuclear wave functions used in the calculation are solutions to the Schroedinger equation with the static one-meson-exchange potential constructed in the respective (sigma) model. The same holds true for the nucler pion production amplitude, since it is proportional to the divergence of the axial current matrix element, by virtue of PCAC. Thus we found a new consistency condition between the pion creation/absorption operator and the nuclear Hamiltonian. We present examples drawn from our models and discuss implications for one-pion-two-nucleon processes.Comment: 19 pages, 7 figures, submitted to Phys. Rev.

Extended Hauser-Feshbach Method for Statistical Binary-Decay of Light-Mass Systems

An Extended Hauser-Feshbach Method (EHFM) is developed for light heavy-ion fusion reactions in order to provide a detailed analysis of all the possible decay channels by including explicitly the fusion-fission phase-space in the description of the cascade chain. The mass-asymmetric fission component is considered as a complex-fragment binary-decay which can be treated in the same way as the light-particle evaporation from the compound nucleus in statistical-model calculations. The method of the phase-space integrations for the binary-decay is an extension of the usual Hauser-Feshbach formalism to be applied to the mass-symmetric fission part. The EHFM calculations include ground-state binding energies and discrete levels in the low excitation-energy regions which are essential for an accurate evaluation of the phase-space integrations of the complex-fragment emission (fission). In the present calculations, EHFM is applied to the first-chance binary-decay by assuming that the second-chance fission decay is negligible. In a similar manner to the description of the fusion-evaporation process, the usual cascade calculation of light-particle emission from the highly excited complex fragments is applied. This complete calculation is then defined as EHFM+CASCADE. Calculated quantities such as charge-, mass- and kinetic-energy distributions are compared with inclusive and/or exclusive data for the $^{32}$S+$^{24}$Mg and $^{35}$Cl+$^{12}$C reactions which have been selected as typical examples. Finally, the missing charge distributions extracted from exclusive measurements are also successfully compared with the EHFM+CASCADE predictions.Comment: 34 pages, 6 Figures available upon request, Phys. Rev. C (to be published

E2F-1 induces melanoma cell apoptosis via PUMA up-regulation and Bax translocation

BACKGROUND: PUMA is a pro-apoptotic Bcl-2 family member that has been shown to be involved in apoptosis in many cell types. We sought to ascertain whether induction of PUMA plays a crucial role in E2F-1-induced apoptosis in melanoma cells. METHODS: PUMA gene and protein expression levels were detected by real-time PCR and Western blot in SK-MEL-2 and HCT116 cell lines after Ad-E2F-1 infection. Activation of the PUMA promoter by E2F-1 overexpression was detected by dual luciferase reporter assay. E2F-1-induced Bax translocation was shown by immunocytochemistry. The induction of caspase-9 activity was measured by caspase-9 colorimetric assay kit. RESULTS: Up-regulation of the PUMA gene and protein by E2F-1 overexpression was detected by real-time PCR and Western blot analysis in the SK-MEL-2 melanoma cell line. In support of this finding, we found six putative E2F-1 binding sites within the PUMA promoter. Subsequent dual luciferase reporter assay showed that E2F-1 expression could increase the PUMA gene promoter activity 9.3 fold in SK-MEL-2 cells. The role of PUMA in E2F-1-induced apoptosis was further investigated in a PUMA knockout cell line. Cell viability assay showed that the HCT116 PUMA-/- cell line was more resistant to Ad-E2F-1-mediated cell death than the HCT116 PUMA+/+ cell line. Moreover, a 2.2-fold induction of the PUMA promoter was also noted in the HCT116 PUMA+/+ colon cancer cell line after Ad-E2F-1 infection. Overexpression of a truncated E2F-1 protein that lacks the transactivation domain failed to up-regulate PUMA promoter, suggesting that PUMA may be a transcriptional target of E2F-1. E2F-1-induced cancer cell apoptosis was accompanied by Bax translocation from the cytosol to mitochondria and the induction of caspase-9 activity, suggesting that E2F-1-induced apoptosis is mediated by PUMA through the cytochrome C/Apaf-1-dependent pathway. CONCLUSION: Our studies strongly demonstrated that E2F-1 induces melanoma cell apoptosis via PUMA up-regulation and Bax translocation. The signaling pathways provided here will further enhance insights on the mechanisms of E2F-1-induced cancer cell apoptosis as a strategy for cancer therapy

Control de cambios / Rastrea los cambios / El camino cambia: Reflexiones sobre un mundo en transformación

“Track changes: Reflecting on a transforming world” was the theme chosen to invite panels, papers, posters and alternative presentations to be part of the 2019 international congress of SIEF that was held in Santiago de Compostela, Galicia (Spain). This introduction includes a description of the content of the congress, the rationale of the choice of plenaries and some reflections about the outcomes of the congress.El lema elegido para presentar paneles, ponencias, posters y presentaciones en formatos alternativos para el congreso internacional 2019 de SIEF -que tuvo lugar en Santiago de Compostela, Galicia (España)- fue “Track changes: Reflecting on a transforming world”. Esta introducción incluye una descripción del contenido del congreso, la idea para la elección de las plenarias y algunas reflexiones sobre los resultados del congreso. &nbsp