Single-strand conformational polymorphism analysis of the M(r) 170,000 isozyme of DNA topoisomerase II in human tumor cells

Five cell lines selected for resistance to the cytotoxicity of inhibitors of DNA topoisomerase II have point mutations in the gene that codes for the M(r) 170,000 form of this enzyme. In each case, the mutation results in an amino acid change in or near an ATP binding sequence of the M(r) 170,000 isozyme of topoisomerase II. We used single-strand conformational polymorphism analysis to screen for similar mutations in other drug-resistant cell lines or in leukemic cells from patients previously treated with etoposide or teniposide. We also analyzed the region of the gene that codes for amino acids adjacent to the tyrosine at position 804 of topoisomerase II which binds covalently to DNA. CEM/VM-1, CEM/VM-1-5, and HL-60/AMSA human leukemic cell lines were used as controls; 3 of 3 known mutations were detected by migration differences of polymerase chain reaction products from the RNA extracted from these three lines. A previously unknown mutation was found in the tyrosine 804 region of the M(r) 170,000 topoisomerase II expressed by CEM/VM-1 and CEM/VM-1-5 cells. Sequence analysis showed that substitution of a T for a C at nucleotide 2404 resulted in an amino acid change of a serine for a proline at amino acid 802. No mutations in any of the ATP binding sequences or in the tyrosine 804 region were detected in polymerase chain reaction products from RNA extracted from human leukemia HL-60/MX2 or CEM/MX1 cells (both cell lines selected for resistance to mitoxantrone) or in human myeloma 8226/Dox1V cells (selected for resistance by simultaneous exposure to doxorubicin and verapamil). No mutations were detected in polymerase chain reaction products from RNA extracted from blasts of 15 patients with relapsed acute lymphocytic leukemia, previously treated with etoposide or teniposide. We conclude that: (a) single-strand conformational polymorphism analysis is useful for screening for mutations in topoisomerase II; (b) resistance to the cytotoxicity of inhibitors of DNA topoisomerase II is not always associated with mutations in ATP binding sequences or the active site tyrosine region of M(r) 170,000 topoisomerase II; and (c) mutations similar to those detected in drug resistant cells selected in culture have not been identified in blast cells from patients with relapsed acute lymphocytic leukemia, previously treated with etoposide or teniposide.     

In Search of the "Ideal" Pear (pyrus spp.): Results of a Multidisciplinary Exploration

Preference mapping with 10 pears identified 3 segments among New Zealand consumers. While 1 segment preferred ripe European cultivars, another rejected only the seedling with strong off‐flavors. A 3rd segment combined the 2 former segments, but also rejected 2 hybrid samples. Survey responses to the question “To me, the ideal pear is ?” from a 2nd consumer sample confirmed the general preference for juicy and sweet pears, the key characteristics of ripeness. A separate appearance evaluation revealed 4 segments based on color and shape. Combining information from several sources resulted in suggestions for new breeding directions for pears, while highlighting the importance of appearance and the need for more extensive measurements.     

Ionospheric wave spectrum measurements

A significant quantity to be measured in the ionosphere is the local spectrum, S(k,ω), of either potential or electron density fluctuations. One can determine from S(k,ω) characteristics of the macroscopic plasma such as the local density and temperature, transport coefficients, and drift current. Determination of S(k,ω) may be carried out by measurement of the cross-power spectrum. Signals from two probes are passed through a cross-power spectrum analyzer, and the measurement is repeated for a series of probe separations. The resulting cross-power spectra, H(r,ω), must be Fourier-transformed to obtain S(k,ω). A number of questions pertinent to the use of this method in space are discussed. These include the expected signal-to-noise ratio, and the frequency and wave-number resolution attainable.     

Attaching community psychiatric nurses to general practice

A scheme for attachment of psychiatric nurses from hospital to group practice in Oxford is described. One community psychiatric nurse can work satisfactorily with eight general practitioners covering a population of about 18,000. From analysis of a working year, it is concluded that this arrangement improves the care of patients in the community by providing psychiatric help at times when it was previously unavailable or unacceptable.The implications of such a scheme for the workings of the primary health care team and the hospital psychiatric service are considered, and a case is made for a further study involving a comparison between practices with and without the attachment of a community psychiatric nurse.     

Dynamic range improvements and measurements in radar systems

The extension of the dynamic range performance of existing radar systems is integral to the requirement for current sensors to see deeper into clutter, that is, to detect weak signals (e.g. small targets) in the presence of strong interference (e.g. urban clutter signals). The rationale behind this requirement is that this additional capability greatly enhances the utility of military and commercial systems. Two approaches to improving the dynamic range of modern radar and surveillance receivers are described: (1) linearisation of intermodulation distortion (IMD) using digital post-distortion algorithms and (2) linearisation of IMD using frequency retranslation mixer (FRM). The experimental results presented illustrate that the digital post-distortion and FRM approaches are effective in providing IMD improvements for the hardware and test scenarios measured. The improvements measured ranged from 15 to 35 dB for narrowband signals to 2-6 dB for frequency-modulated continuous wave (FMCW) wideband signals. The digital post-distortion approach provided the greatest performance improvement when the input signals were between the analogue-to-digital converter full range and 20 dB below that level. The improvements measured for the FRM linearisation approach ranged from 20 to 25 dB for the narrowband two-tone tests and 16-18 dB for the wideband FMCW signal chirps.     

Determining consumer purchase intentions: the importance of dry matter, size, and price of kiwifruit

Abstract: Knowledge of the relative importance of food quality attributes in determining consumer purchase intention is critical for robust assessment of economic opportunities for industry growth. The aim of this study is to demonstrate how conjoint analysis methodology that incorporates tasting of fruit can be used to collect such information. Three hundred Japanese consumers took part in research designed to measure the importance of dry matter (DM), size, and price of kiwifruit (Actinidia deliciosa “Hayward” and Actinidia chinensis “Hort16A”) for purchase intention. Measurement of consumer liking for kiwifruit of different DM content was a key first step. Liking increased as DM increased and was accompanied by increased purchase likelihood/choice probability for kiwifruit. The size of kiwifruit presented to consumers varied from “small” to “extra large.” Consumers liked “mid‐sized” kiwifruit over “small” or “extra‐large” kiwifruit. Despite these differences in liking, size was of little importance in determining purchase likelihood/choice probability for kiwifruit. Price was a very important factor in determining purchase likelihood/choice probability but was less important than DM content. As price increased, purchase likelihood/choice probability decreased. Beneath these general findings, heterogeneity existed. Some consumers placed more/less importance on the focal purchase drivers than suggested by the aggregate model. Overall, the results suggest that incentive schemes already implemented by industry should consider rewarding high‐DM fruit more than fruit size. Practical Application: This research has contributed to the New Zealand kiwifruit industry gaining a better understanding of the relative importance consumers place on DM, size, and price of kiwifruit and has resulted in changes to grower incentive schemes. The research approach presented forces consumer to tradeoff attributes of kiwifruit against each other and decide on how important two key quality attributes—DM and size—are relative to each other and relative to price. The application of conjoint analysis in this article can be transferred to other fruits, food, and beverages and help guide consumer‐led innovation.     

Microstructures of Simulated Nuclear Waste

The microstructures of simulated tailored ceramic forms for the Idaho Chemical Processing Plant (ICPP) high-level waste were characterized by TEM. The ceramic forms were consolidated from simulated ICPP waste calcines with either silicayttria-based or silica–lithia-based additives by hot isostatic pressing. The hot isostatically pressed ceramic waste forms are composed of cubic CaF₂, monoclinic ZrO₂, stabilized cubic ZrO₂, tetragonal ZrSiO₄, and amorphous silicate phases which can be phase separated. The phaseseparated glass is a result of phase immiscibility in the soda aluminosilicate glass to the compositions of albite and mullite.     

Antithrombotic potency of hirudin is increased in nonhuman primates by fibrin targeting

BACKGROUND: Inhibition of thrombin by either the indirect thrombin inhibitor heparin or by more potent direct thrombin inhibitors such as hirudin reduces thrombus formation after arterial injury. The present study was designed to determine if a fibrin-specific thrombin inhibitor could, by local thrombin inhibition, prevent thrombosis more effectively. METHODS AND RESULTS: We first studied antithrombotic potency in vitro, comparing fibrin-targeted hirudin (recombinant hirudin covalently linked to the Fab' fragment of the anti-fibrin monoclonal antibody 59D8) to recombinant hirudin in baboon plasma. Fibrin-targeted hirudin was nine times more effective than recombinant hirudin in inhibiting fibrin deposition on experimental clot surfaces in baboon plasma (P < .01). The potency of fibrin-targeted hirudin was then compared with that of recombinant hirudin in a baboon model of thrombus formation. 111In-labeled platelet deposition was measured in a synthetic graft segment of an extracorporeal arteriovenous shunt in control animals and in animals receiving either fibrin-targeted hirudin or hirudin. In these experiments, fibrin-targeted hirudin was 10-fold more potent than hirudin in inhibiting platelet deposition and thrombus formation (P < .05). CONCLUSIONS: These data indicate that targeting a thrombin inhibitors such as hirudin to an epitope present in thrombi results in increased antithrombotic potency.