Antisense oligonucleotide inhibition of hepatitis C virus gene expression in transformed hepatocytes

Genetic and biochemical studies have provided convincing evidence that the 5' noncoding region (5' NCR) of hepatitis C virus (HCV) is highly conserved among viral isolates worldwide and that translation of HCV is directed by an internal ribosome entry site (IRES) located within the 5' NCR. We have investigated inhibition of HCV gene expression using antisense oligonucleotides complementary to the 5' NCR, translation initiation codon, and core protein coding sequences. Oligonucleotides were evaluated for activity after treatment of a human hepatocyte cell line expressing the HCV 5' NCR, core protein coding sequences, and the majority of the envelope gene (E1). More than 50 oligonucleotides were evaluated for inhibition of HCV RNA and protein expression. Two oligonucleotides, ISIS 6095, targeted to a stem-loop structure within the 5' NCR known to be important for IRES function, and ISIS 6547, targeted to sequences spanning the AUG used for initiation of HCV polyprotein translation, were found to be the most effective at inhibiting HCV gene expression. ISIS 6095 and 6547 caused concentration-dependent reductions in HCV RNA and protein levels, with 50% inhibitory concentrations of 0.1 to 0.2 microM. Reduction of RNA levels, and subsequently protein levels, by these phosphorothioate oligonucleotides was consistent with RNase H cleavage of RNA at the site of oligonucleotide hybridization. Chemically modified HCV antisense phosphodiester oligonucleotides were designed and evaluated for inhibition of core protein expression to identify oligonucleotides and HCV target sequences that do not require RNase H activity to inhibit expression. A uniformly modified 2'-methoxyethoxy phosphodiester antisense oligonucleotide complementary to the initiator AUG reduced HCV core protein levels as effectively as phosphorothioate oligonucleotide ISIS 6095 but without reducing HCV RNA levels. Results of our studies show that HCV gene expression is reduced by antisense oligonucleotides and demonstrate that it is feasible to design antisense oligonucleotide inhibitors of translation that do not require RNase H activation. The data demonstrate that chemically modified antisense oligonucleotides can be used as tools to identify important regulatory sequences and/or structures important for efficient translation of HCV.     

Comparison of PhD programs in clinical and counseling psychology

Some specific characteristics of the aging of the Brazilian population in different areas, states and communities all over the country, have shown significant variations. Historical series of demographic and health indicators for the population in their sixties and over in Brazil, state of S. Paulo and in the municipal district of Araraquara are listed as follows: level of education and urban population growth rate, income distribution, mortality rates and main causes of death. In 1991 the aged constituted were 7.8% of the Brazilian population and 9.7% in Araraquara community. The elderly population (of 70 years of aged and above) as a proportion of the whole, has increased and already stands for 40%. The same trend holds good for both the proportion of aged within the urban population and their level of education which increased to 90% in 1991. The main causes of death are chronic degenerative diseases which have replaced the infectious illness: first, the diseases of the circulatory system (which account for more than 40% of all deaths) and the neoplasms (which let to 15% of the deaths). On the basis of these health and demographic data relating to people of 60 years of age and over, this study suggests some procedures for the improvement of the quality of the assistance given to the target population: a) the assistance give to the aged should be improved by providing gerontological training for general physicians and nurses, both of public and private clinics; b) the already existing educational activities for the aged, for health workers and for teachers of secondary education should be further developed; c) the number of day-hospitals should be increased for the purpose of avoiding unnecessary confinement so as maintain the low rate of institutionalization in homes for the elderly (0.7% in Araraquara). It is reported that at least 35% of the aged population in this area is entitled to private health assistance, which brings out the importance of including such services in the local health programs for this group.     

High brachial artery bifurcation: a report of 2 cases

High division of the brachial artery was observed in two cadavers, during routine dissection of upper extremities. In the first case, the brachial artery of the right upper extremity was divided into its two terminal branches immediately after passing between the lateral and medial roots of the median nerve and just below the origin of the profunda brachii artery. The lateral branch was the radial artery, located in the space normally occupied by the brachial artery and the medial one was the ulnar artery. In the second case, the brachial artery was divided into its two terminal branches just below the origin of the profunda brachii artery. Accurate knowledge of the relationships and course of these major arterial conduits and particularly of their variational patterns, is of considerable practical importance in the conduct of reperative surgery in the arm, forearm and hand.     

Sulfated zirconia catalysts: Are Brønsted acid sites the source of the activity?

The thermal decomposition products of pyridinium sulfate differ from those of pyridinium sulfate supported on zirconia which in turn differs from that of pyridine adsorbed on a sulfated zirconia. Unsupported pyridinium sulfate decomposes to produce pyridine and sulfuric acid, and these subsequently react to produce oxides of carbon and sulfur. Zirconia that is sulfated and then exposed to pyridine does not release detectable amount of pyridine during heating in an inert gas; rather the pyridine undergoes oxidation reduction reactions simultaneously to release CO₂ and sulfur compounds. Pyridinium sulfate supported on zirconia decomposes upon heating to release pyridine and sulfuric acid, which reacts with the zirconia. The desorption of pyridine in one case and only CO₂/SO_x in the other case suggests that sulfated zirconia does not contain Brønsted acidity that can form pyridinium sulfate.     

Process monitoring using auto-associative,feed-forward artificial neural networks

The potential of using artificially simulated neural networks as intelligent, adaptive process-monitoring devices is discussed. The investigation is considered as a method for automatic, intelligent exception reporting for quality control applications. The technique is also compared with the conventional statistical approaches of principal component analysis and Kohonen's feature map. The applications of the technique in aerospace and manufacturing environments are presented and a possible extension of the method to incorporate a diagnostic function is discussed.Seconded from Cheltenham and Gloucester College of Higher Education as a Royal Society/SERC Research Fellow at Smith's Industries Aerospace and Defence Systems, Bishop's Cleeve, Cheltenham, UK.     

Current practice in software quality and the impact of certification schemes

The paper presents the findings of a survey that investigated the level of quality management practice within some 150 UK companies from a sample of 500. It provides a snapshot of practice at the time of the survey, and assesses the impact of government quality initiatives particularly, the TickIT scheme at that time. The survey methodology is described, together with the results and conclusions. The sample has been graded by size of company, which the authors consider to have a significant effect upon the adoption of quality practices. The survey highlights the need to encourage small companies to adopt quality practices and to assist them with the short-term costs incurred.     

Improvements to an electromagnetic near-field sensor forsimultaneous electric and magnetic field measurement

A sensor for simultaneous electric and magnetic field measurement has been described by M. Kanda (see ibid., vol.EMC-26, no.3, p.102-10, 1984). It relies upon measuring the voltage across precisely matched loads at opposite sides of a loop antenna. A modification to the theory is presented to account for unmatched loading of the loop. Measured results of wave impedance over a 10-400-MHz range demonstrate the accuracy of the modified theory as well as highlighting a novel calibration technique     

1.3 μm InAsyP1-y-InPstrained-layer quantum-well laser diodes grown by metalorganic chemicalvapor deposition

The authors have fabricated 1.3-μm InAsP-InP separate-confinement-heterostructure (SCH) strained-layer double-quantum-well (SL-DQW) laser diodes (LDs) by metalorganic chemical vapor deposition (MOCVD). A low threshold current density of 410 A/cm ² was obtained. The CW threshold current was as low as 1.8 mA at 20°C, and maximum CW operating temperature of 120°C was obtained. These characteristics are almost the same as those of well-designed GaInAsP-InP SL-QW LDs. Further improvement of the characteristics of InAsP-InP LDs is expected by optimizing the device structure     

The correlation of texture and magnetic properties in Co-Cr films

This investigation focuses on the correlation between the crystallographic orientation of grains with respect to magnetic properties in Co-Cr films. Based on a distribution measured for the (0002) crystallographic pole densities, modified formulae describing the texture were used to calculate the coercivity Hc, the squareness ratio Rs (Mr⊥/TMs) and the anisotropy field Ha for Co-Cr films. In general, calculated values for the coercive force He agreed well with the measured values. The calculated Rs⊥. values were much larger than the measured ones, however, indicating that the influence of the demagnetizing field and the magnetostriction on the orientation of the magnetization cannot be neglected. On leave from Institute of Computing Technology, Academia Sinica, P.O.Box 2704-6, Beijing, P.R. China.