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

Effect of River Levee with Geosynthetic-Reinforced Soil against Overflow Erosion and Infiltration

Overflows from huge floods have caused levee breaches in a great number of places, including Japan. To prevent such destruction and thereby increase the resistance of armored levees to overflow erosion, in this study, we examined the performances of Geosynthetic-Reinforced Soil (GRS) levees against overflow erosion under various conditions, such as reinforcement, back slopes, and geo-grid layers. In addition, we investigated the effect of geo-grid layers on the infiltration of levees. The model tests revealed that 1) with scour protection in front of the toe of the back slope, the GRS levee exhibits much higher resistance against overflow erosion than the armored levee; 2) the armored levee with a steep back slope (= 1:0.5) collapsed faster than that with a normal slope (= 1:2). However, the GRS levee with a steep back slope of 1:0.5 maintained high resistance against overflow erosion after the target time. 3) The GRS levee with partial and full reinforcements had a comparably high resistance against overflow erosion. 4) The GRS levee using a small-sized geo-grid maintained a high residual ratio of the cross-sectional area over a long period. 5) The infiltration discharge of the GRS levee was lesser than that of the levee with no reinforcement due to the reduction in infiltration erosion in the GRS levee. These facts suggest that the GRS levee with partial reinforcement can be applied to the reinforcement of existing levees, and appropriately sized geo-grid layers should be selected

The factors associated with pain severity in patients with knee osteoarthritis vary according to the radiographic disease severity: a cross-sectional study

SummaryObjectivesKnee osteoarthritis (OA) pain is suggested to be associated with inflammation and detrimental mechanical loading across the joint. In this cross-sectional study, we simultaneously examined the inflammation and alignment of the lower limb and examined how the pain components varied depending on the disease progression.DesignOne-hundred sixty female medial type of early- [n = 74 in Kellgren–Lawrence (K/L) 2] to advanced-stage (n = 96 in K/L >2) knee OA subjects (70.5 years on average) were enrolled. Knee pain was evaluated using a pain visual analog scale (VAS) and the pain-related subcategory of the Japanese Knee Osteoarthritis Measure (JKOM-pain). The serum interleukin (sIL)-6 level reflecting synovitis, and the high sensitivity C-reactive protein (hs-CRP) level were measured to evaluate the severity of inflammation. The anatomical axis angle (AAA) was measured as an alignment index. The β-coefficient was estimated after adjusting for age and the body mass index (BMI) using a multiple linear regression analysis.ResultsMultiple linear regression analyses showed that the sIL-6 levels, but not AAA, associated with the pain VAS [β = 10.77 (95% confidence interval (CI): 4.14–17.40), P < 0.01] and JKOM-pain scores [β = 3.19 (95% CI: 1.93–4.44), P < 0.001] in the early stage. Conversely, AAA, but not the sIL-6 levels, was found to be associated with the pain VAS [β = −1.29 (95% CI: −2.51 to −0.08), P < 0.05] and JKOM-pain scores [β = −0.49 (95% CI: −0.82 to −0.16), P < 0.01] in the advanced stage.ConclusionsThe presence of a higher level of sIL-6 and the varus alignment of the joint is associated with pain in early- and advanced-stage knee OA patients, respectively

The degeneration and destruction of femoral articular cartilage shows a greater degree of deterioration than that of the tibial and patellar articular cartilage in early stage knee osteoarthritis: a cross-sectional study

SummaryObjectiveThe aim of the present study was to examine whether the degenerative and morphological changes of articular cartilage in early stage knee osteoarthritis (OA) occurred equally for both femoral- and tibial- or patellar- articular cartilage using magnetic resonance imaging (MRI)-based analyses.DesignThis cross-sectional study was approved by the ethics committee of our university. Fifty patients with early stage painful knee OA were enrolled. The patients underwent 3.0 T MRI on the affected knee joint. Healthy volunteers who did not show MRI-based OA changes were also recruited as controls (n = 19). The degenerative changes of the articular cartilage were quantified by a T2 mapping analysis, and any structural changes were conducted using Whole Organ Magnetic Resonance Imaging Score (WORMS) technique.ResultsAll patients showed MRI-detected OA morphological changes. The T2 values of femoral condyle (FC) (P < 0.0001) and groove (P = 0.0001) in patients with early stage knee OA were significantly increased in comparison to those in the control, while no significant differences in the T2 values of patellar and tibial plateau (TP) were observed between the patients and the control. The WORMS cartilage and osteophyte scores of the femoral articular cartilage were significantly higher than those in the patellar- (P = 0.001 and P = 0.007, respectively) and tibial- (P = 0.0001 and P < 0.0001, respectively) articular cartilage in the patients with early stage knee OA.ConclusionsThe degradation and destruction of the femoral articular cartilage demonstrated a greater degree of deterioration than those of the tibial- and patellar- articular cartilage in patients with early stage knee OA

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

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

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