Differential formation of hydroxyl radicals by adriamycin in sensitive and resistant MCF-7 human breast tumor cells: implications for the mechanism of action

Adriamycin-stimulated formation of .OH in sensitive and resistant subline of human breast tumor cells (MCF-7) has been examined by electron spin resonance spectroscopy. It was shown that adriamycin significantly stimulated the formation of .OH spin adducts [5,5-dimethyl-1-pyrroline N-oxide (DMPO)-OH] in the sensitive cells but not in the resistant cells. By use of spin-broadening techniques and inhibition of .OH with high molecular weight poly(ethylene glycol), which does not enter intact cells, it was shown that 60-65% of adriamycin-induced .OH were located extracellularly and were metal ion dependent since they were decreased in the presence of desferal. Furthermore, superoxide dismutase and catalase, enzymes that detoxify superoxide and hydrogen peroxide, also significantly inhibited adriamycin-induced .OH formation and protected against the cytotoxicity of adriamycin. The differential .OH formation in these two cell lines is not due to diminished activities of flavin-dependent activating enzymes nor decreased accumulation of the drug in the cells but appears to be related to enhanced activities of detoxifying enzymes, particularly, glutathione peroxidases in the resistant cells.     

Statistical analysis of drug-drug and site-site interactions with isobolograms

The use of more than one drug to achieve a desired effect has been a common practice in pharmacologic testing and in clinical practice. For example, combinations of analgesics are frequently prescribed with a view to enhancing pain relief and reducing adverse effects. It is also well established that administration of more than one drug may give effects that are greater than, or less than, the additive effect of each drug given individually. A non-mechanistic method of characterizing the effect resulting from the administration of two compounds is the isobologram. It is relatively simple to draw and interpret isobolograms. However, this graphical technique, which employs equieffective concentrations of individual drugs and combinations of these, obtains the concentrations as random variables from concentration-effect data, usually transformed to a parallel line assay. Thus, statistical confidence limits from such assays, as well as from non-parallel designs, must be expressed on the isobologram if this diagram is to establish superadditive, subadditive, or merely additive effects. We now present a detailed statistical analysis of the isobolographic method illustrated with examples of the statistical procedures, a rational basis for selecting proportions of each drug in the combination, and a relatively novel application of the isobolographic concept, i.e., interactions involving different anatomical sites.     

Using California Gnatcatcher to Test Underlying Models in Habitat Conservation Plans

Abstract: Habitat Conservation Plans are a widely used strategy to balance development and preservation of species of concern and have been used in southern California, USA, to protect the coastal California gnatcatcher (Polioptila californica). Few data exist on gnatcatcher abundance and distribution, and existing data have problems with issues of closure (i.e., sampling occurs in a short enough time period such that abundance or distribution are not changing), detectability, and proper attention to probability-based sampling schemes. Thus, a habitat model has been relied upon in reserve design. California gnatcatchers are the flagship and umbrella species of many plans and we provide the first estimates that incorporate probabilistic sampling and test predictions from the habitat model. Probability of occurrence was 26% (SĚ = 0.06); however, occupancy varied by modeled habitat quality with slopes <40%, warm, and wet sagebrush habitat having higher occupancy probabilities. Interpreting abundance and occupancy probabilities by vegetation type was complicated by error detected in Geographic Information System vegetation metadata files. The slope (1.08, SĚ = 0.66), temperature (0.79, SĚ = 0.70), and precipitation (—2.62, SĚ = 1.21) variables associated with habitat models were stronger influences on occupancy than was patch size (0.48, SĚ = 0.66). Previous models weight patch size equal to slope and climate. Our work demonstrates that probabilistic sampling can be carried out on a large scale and the results provide better information for managers to make decisions about their reserves.     

New alleles of IGKV genes A2 and A18 suggest significant human IGKV locus polymorphism

The human kappa light chain consists of approximately 35 potentially functionalIGKV genes. However, an estimation of the diversity in theIGKV repertoire of an individual will be affected by the extent of polymorphisms for the differentIGKV genes and their patterns of inheritance. To date, little information is available to indicate the extent of allelic variation of theIGKV genes. We examined the extent of allelism for oneIGKV gene pair, the distal regionA2 gene and its closely related proximal region duplicateA18. We found two new alleles forA2 and one new allele forA18, and sequenced ∼ 1 kilobase flanking each gene. The newA18 allele, unlike the originally described allele, appears to be functional. All these alleles were found at relatively high frequencies in the four ethnic populations studied, with the exception of the defectiveA2b allele which was highly represented only in Navajos. The originally describedA2a allele encodes for the predominant protective antibody againstHaemophilus influenzae. Therefore, the patterns of allelic inheritance described for thisIGKV gene pair indicate that allelism in theIGKV locus is likely to have a significant impact on immune responses.     

In vivo studies with ICI 154,129, a putative delta receptor antagonist

We studied the in vivo pharmacology of ICI 154,129, a new antagonist that is claimed to show selectivity for delta opiate receptors. At s.c. doses of 30 and 100 mg/kg, ICI 154,129 had no marked effect on the gastrointestinal transit of a charcoal meal in mice. In this test, ICI 154,129 reversed the inhibitory action of metkephamid (a proposed delta receptor agonist) but not that of levorphanol. ICI 154,129 was proconvulsant in the mouse picrotoxin potentiation test; the dose-response curve had a low ceiling and was biphasic. Naloxone (1 mg/kg, s.c.) enhanced the proconvulsant action of ICI 154,129 (40 mg/kg, s.c.) by an unknown mechanism.     

Anti-Rous sarcoma response of major histocompatibility (B) complex haplotypes B23, B24 and B30

Responses to Rous sarcoma virus (RSV) induced tumours were studied in UNH 105, a non-inbred line of New Hampshire chickens. Six single male matings encompassing a total of 50 dams produced 345 progeny which segregated for B complex genotypes B23/B23, B23/B24, B23/B30, B24/B24, B24/B30 and B30/B30. Six-week-old chicks were wingweb inoculated with a pseudotype of Bryan high titre Rous sarcoma virus, BH RSV (RAV-1). Tumours were scored for size six times over a 10-week period post-inoculation. Each chick was assigned a tumour profile index (TPI) as an indicator of immunological response. The number of days to death (DTD) was recorded for 148 chicks with terminal tumours. Genotypes B23/B23, B23/B24 and B23/B30, with TPIs of 1.8, 1.7 and 2.0 respectively, did not differ significantly from each other, suggesting dominance of response of B23 over B24 and B30 haplotypes. B24/B30 chicks with the highest TPI (3.4) and shortest DTD (34.6) were significantly different from B30/B30 (2.8; 41.6) but not from B24/B24 (3.1; 34.9) suggesting dominance of response of the B24 haplotype over B30 in the absence of B23.