Evaluation of bacterial RNase P RNA as a drug target

RNA has gained increasing importance as a therapeutic target. However, so far mRNAs rather than stable cellular RNAs have been considered in such studies. In bacteria, the tRNA-processing enzyme RNase P has a catalytic RNA subunit. Fundamental differences in structure and function between bacterial and eukaryotic RNase P, and its indispensability for cell viability make the bacterial enzyme an attractive drug target candidate. Herein we describe two approaches utilized to evaluate whether the catalytic RNA subunit of bacterial RNase P is amenable to inactivation by antisense-based strategies. In the first approach, we rationally designed RNA hairpin oligonucleotides targeted at the tRNA 3'-CCA binding site (P15 loop region) of bacterial RNase P RNA by attempting to include principles derived from the natural CopA-CopT antisense system. Substantial inactivation of RNase P RNA was observed for Type A RNase P RNA (such as that in Escherichia coli) but not for Type B (as in Mycoplasma hyopneumoniae). Moreover, only an RNA oligonucleotide (Eco 3') complementary to the CCA binding site and its 3' flanking sequences was shown to be an efficient inhibitor. Mutation of Eco 3' and analysis of other natural RNase P RNAs with sequence deviations in the P15 loop region showed that inhibition is due to interaction of Eco 3' with this region and occurs in a highly sequence-specific manner. A DNA version of Eco 3' was a less potent inhibitor. The potential of Eco 3' to form an initial kissing complex with the P15 loop did not prove advantageous. In a second approach, we tested a set of oligonucleotides against E. coli RNase P RNA which were designed by algorithms developed for the selection of suitable mRNA targets. This approach identified the P10/11-J11/12 region of bacterial RNase P RNA as another accessible region. In conclusion, both the P15 loop and P10/11-J11/12 regions of Type A RNase P RNAs seem to be promising antisense target sites since they are easily accessible and sufficiently interspersed with nonhelical sequence elements, and oligonucleotide binding directly interferes with substrate docking to these two regions.     

Apoptosis in interface membranes of aseptically loose total hip arthroplasty

The terminal events leading to periprosthetic osteolysis are multifactorial in nature and modulation of this process after the stage of osteolytic mediator release has been futile. Recently, the demonstration of the ability of bisphosphonates to inhibit bone resorption that is mediated by particle-stimulated macrophages and their induction of osteoclast apoptosis suggests a potent area for modulation of osteolysis at the prosthesis-bone interface. The purpose of this study was to determine the mode of cell death that occurs at the osteolytic interface of failed total hip arthroplasty (THA). TUNEL staining, DNA laddering, and immunodetection of poly(ADP-ribose)polymerase (PARP) protein were used to identify the presence of apoptosis in interface membranes from 25 patients aged 28–88 years old (mean, 58 years) harvested at the time of hip revision surgery. Our results demonstrated positive TUNEL stain in 100% of specimens with an average 37% of cells (range 12–60%) positively stained for TUNEL whereas less than 8% of control tissue cells showed positive staining. DNA laddering, a characteristic feature of apoptotic cells, was observed in 82% (28/34) of specimens studied at both the acetabular and femoral side of aseptically loose THAs. No laddering was observed in control tissues. Finally, using Western blot analysis, we observed the appearance of the 89 kDa PARP fragment associated with apoptosis in 92% of specimens (30/33). Our results demonstrate the presence of apoptotic cell death in interface membranes of THAs suggesting that apoptosis-related events are indeed associated with periprosthetic osteolysis and could serve as a specific target point for therapeutic modulation. © 2001 Kluwer Academic Publishers     

Temperature‐Dependent Exciton and Trap‐Related Photoluminescence of CdTe Quantum Dots Embedded in a NaCl Matrix: Implication in Thermometry

Temperature‐dependent optical studies of semiconductor quantum dots (QDs) are fundamentally important for a variety of sensing and imaging applications. The steady‐state and time‐resolved photoluminescence properties of CdTe QDs in the size range from 2.3 to 3.1 nm embedded into a protective matrix of NaCl are studied as a function of temperature from 80 to 360 K. The temperature coefficient is found to be strongly dependent on QD size, with the highest sensitivity obtained for the smallest size of QDs. The emission from solid‐state CdTe QD‐based powders is maintained with high color purity over a wide range of temperatures. Photoluminescence lifetime data suggest that temperature dependence of the intrinsic radiative lifetime in CdTe QDs is rather weak, and it is mostly the temperature‐dependent nonradiative decay of CdTe QDs which is responsible for the thermal quenching of photoluminescence intensity. By virtue of the temperature‐dependent photoluminescence behavior, high color purity, photostability, and high photoluminescence quantum yield (26%–37% in the solid state), CdTe QDs embedded in NaCl matrices are useful solid‐state probes for thermal imaging and sensing over a wide range of temperatures within a number of detection schemes and outstanding sensitivity, such as luminescence thermochromic imaging, ratiometric luminescence, and luminescence lifetime thermal sensing.     

Proplant: Multiagent system for production planning

The so-called ''tribase'' acquaintance model of the agent's behavior is presented in this paper. This represents an extension of the twin-base model (Cao et al., 1997). Based on practical experience, the new model tries to cope with parallel processing, precedence constraints, and sparse resources. The idea of substituting the interagent negotiation processes by the periodical internal planning activity of the agents is stressed. A multiagent system, ProPlanT, as an application of the tribase model for the project-oriented production planning developed for TESLA TV company is described in detail. Three types of agents production planning agent (PPA), production management agent (PMA), and production agent (PA) are distinguished. The corresponding tribase models and potential role of metaagents are discussed.     

Applications of real number theorem proving in PVS

Real number theorem proving has many uses, particularly for verification of safety critical systems and systems for which design errors may be costly. We discuss a chain of developments building on real number theorem proving in PVS. This leads from the verification of aspects of an air traffic control system, through work on the integration of computer algebra and automated theorem proving to a new tool, NRV, first presented here that builds on the capabilities of Maple and PVS to provide a verified and automatic analysis of Nichols plots. This automates a standard technique used by control engineers and greatly improves assurance compared with the traditional method of visual inspection of the Nichols plots.     

Microbial and enzymatic stability of fruit juice-milk beverages treated by high intensity pulsed electric fields or heat during refrigerated storage

The influence of high intensity pulsed electric fields (HIPEF) treatment (35 kV/cm, bipolar 4 μs square wave pulses at 200 Hz) on the inactivation of Listeria innocua in fruit juice–whole (FJ–WM) or skim milk (FJ–SM) beverages was assessed. As well, the effects of HIPEF or conventional thermal (90 °C 60 s) processing in terms of microbial and enzymatic stability and changes on physicochemical parameters of the beverages during refrigerated storage were also studied. HIPEF and thermal processing ensured the microbial stability of the beverages during 56 days at 4 °C without significant changes on pH, acidity and soluble solid content values. Thermal processing was more effective than HIPEF in reducing Pectin Methyl Esterase (PME) activity in FJ–SM and FJ–WM beverages since its activity was reduced by 70.66% and 87.33%, respectively. With regards to Polygalacturonase (PG), HIPEF treatment reduced by 26.92% and 20.93% the enzyme activity in the FJ–WM and FJ–SM beverages, respectively, whereas thermal treatment did not inactivate PG in both beverages. HIPEF or thermally-treated samples showed higher viscosity than the untreated ones, being this increment more pronounced in the beverages with whole milk. Therefore, HIPEF processing may be a feasible technology in order to obtain shelf stable and fresh-like fruit juice-milk beverages.     

Soft Repulsion and the Behavior of Equations of State at High Pressures

The so-called characteristic curves of Brown—the Amagat (Joule inversion), Boyle, and Charles (Joule–Thomson inversion) curves—of hydrogen are calculated with several equations of state. This work demonstrates that not all equations can generate physically reasonable Amagat curves. After inclusion of corrections for soft repulsion (based on the Weeks–Chandler–Andersen perturbation theory) and quantum effects into the simplified perturbed-hard-chain theory (SPHCT) equation of state, this equation is able to not only generate an Amagat curve, but also predict pVT data, residual Gibbs energies, and heat capacities of several gases at and above 100 MPa reasonably well.     

Inclusion Compounds of Dehydrocholic Acid with Solvents

The host-guest inclusion of various organic solvents within dehydrocholic acid has been studied and the selectivity of enclathration determined by competition experiments.