Reduced drug accumulation and multidrug resistance in human breast cancer cells without associated P-glycoprotein or MRP overexpression

MCF-7 human breast cancer cells selected in Adriamycin in the presence of verapamil developed a multidrug resistant phenotype, which was characterized by as much as 100,000-fold resistance to mitoxantrone, 667-fold resistance to daunorubicin, and 600-fold resistance to doxorubicin. Immunoblot and PCR analyses demonstrated no increase in MDR-1 or MRP expression in resistant cells, relative to parental cells. This phenotype is similar to one previously described in mitoxantrone-selected cells. The cells, designated MCF-7 AdVp, displayed a slower growth rate without alteration in topoisomerase II alpha level or activity. Increased efflux and reduced accumulation of daunomycin and rhodamine were observed when compared to parental cells. Depletion of ATP resulted in complete abrogation of efflux of both daunomycin and rhodamine. No apparent alterations in subcellular daunorubicin distribution were observed by confocal microscopy. No differences were noted in intracellular pH. Molecular cloning studies using DNA differential display identified increased expression of the alpha subunit of the amiloride-sensitive sodium channel in resistant cells. Quantitative PCR studies demonstrated an eightfold overexpression of the alpha subunit of the Na+ channel in the resistant subline. This channel may be linked to the mechanism of drug resistance in the AdVp cells. The results presented here support the hypothesis that a novel energy-dependent protein is responsible for the efflux in the AdVp cells. Further identification awaits molecular cloning studies.     

Changes of respiratory chain activity in mitochondrial and synaptosomal fractions isolated from the gerbil brain after graded ischaemia

In this study we have examined (1) the integrated function of the mitochondrial respiratory chain by polarographic measurements and (2) the activities of the respiratory chain complexes I, II-III, and IV as well as the ATP synthase (complex V) in free mitochondria and synaptosomes isolated from gerbil brain, after a 30-min period of graded cerebral ischaemia. These data have been correlated with cerebral blood flow (CBF) values as measured by the hydrogen clearance technique. Integrated functioning of the mitochondrial respiratory chain, using both NAD-linked and FAD-linked substrates, was initially affected at CBF values of approximately 35 ml 100 g-1 min-1, and declined further as the CBF was reduced. The individual mitochondrial respiratory chain complexes, however, showed differences in sensitivity to graded cerebral ischaemia. Complex I activities decreased sharply at blood flows below approximately 30 ml 100 g-1 min-1 (mitochondria and synaptosomes) and complex II-III activities decreased at blood flows below 20 ml 100 g-1 min-1 (mitochondria) and 35-30 ml 100 g-1 min-1 (synaptosomes). Activities declined further as CBF was reduced below these levels. Complex V activity was significantly affected only when the blood flow was reduced below 15-10 ml 100 g-1 min-1 (mitochondria and synaptosomes). In contrast, complex IV activity was unaffected by graded cerebral ischaemia, even at very low CBF levels.     

Temperamental resistance to control increases the association between sleep problems and externalizing behavior development.

This study examined the moderating effects of temperamental resistance to control on the link between development of sleep problems and development of externalizing behaviors over a 5-year period. Resistance to control was assessed with mothers' retrospective reports of temperament in infancy, provided when children were 5 years of age. Sleep problems were assessed with mother reports on an annual basis from age 5 to age 9. Externalizing behaviors were assessed with teacher reports on an annual basis from age 5 to age 9. A cross-domain latent growth curve model indicated that sleep problem trajectories were positively associated with externalizing behavior trajectories only for children high in resistance to control. In addition, resistance to control was positively associated with initial (age 5) sleep problems and initial (age 5) externalizing behaviors. The authors speculate that the development of sleep problems promotes the development of behavior problems for resistant children, whose self-regulatory abilities are especially tenuous. Implications for prevention and treatment of conduct problems are considered. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Chromosome 9p deletions in invasive and noninvasive nonfunctional pituitary adenomas: the deleted region involves markers outside of the MTS1 and MTS2 genes

We have screened 57 cases of primary, nonfunctional, pituitary adenomas for loss of heterozygosity of markers on chromosome 9p. Using a panel of 11 microsatellite markers, we found hemizygous deletion with at least one of the markers in 18 tumors (31.5%). The frequency of loss was similar in both noninvasive (8 of 26; 31%) and invasive tumors (10 of 31; 32%), suggesting that loss on this chromosome might be an early event in pituitary tumorigenesis. Two discrete areas of loss were punctuated by a region of retention of heterozygosity between the markers D9S171 and IFNA, indicative of homozygous deletion. However, multiplex PCR analysis (MTS1 and MTS2) and the presence of a 3' untranslated region polymorphism in MTS1 suggested that neither of these tumor suppressor genes was homozygously deleted. In 6 of the 18 tumors showing LOH, sufficient DNA was also available for Southern blot analysis and, in all cases, showed retention of MTS1. Cell mixing experiments of tumor cell DNA homozygously deleted for MTS1 with DNA in which neither copy of the gene was deleted only gave rise to a signal at contamination levels greater than 30% and could discriminate homozygous and hemizygous loss. These studies support the recent findings that mechanisms other than hemi- and homozygous deletion are most likely responsible for the loss of MTS1 gene product in pituitary tumors (M. Woloschak et al., Cancer Res., 56: 2493-2486, 1996.). These data show that losses on either side of 9p21-22, both or either of which may be deleted, are involved in pituitary tumorigenesis and provide evidence for distinct suppressor gene loci, in addition to MTS1, on chromosome 9p.     

Determination of synthetic growth promoters in bile

To monitor the use of synthetic growth promoting hormones in red meat animals requires a sensitive and rapid method of analysis. This paper describes the development of such a method for these agents in bile, based on a high pressure liquid chromatographic purification procedure with detection and estimation by radioimmunoassay.     

Distribution of diethylstilboestrol in pigs

The synthetic growth promoter diethylstilboestrol (DES) administered orally to pigs can be detected by radioimmunoassay (RIA) analysis of kidney, liver, faeces, bile and urine from animals fed continuously to slaughter, but not in muscle, fat or plasma. If treated animals are fed on material not containing DES for 72 h prior to slaughter, then the levels of parent compound and metabolite in all products decrease to become not significantly greater than those in control animals. The gross metabolism of DES in pigs and bovines appears to be similar.     

Translational motion compensation for coronary angiogram sequences

A method of compensating for the lag of the video cameras typically used in angiographic systems is presented for use in sequences of digitized X-ray images. The lag effect is reduced by a straightforward weighted subtraction, which has the undesirable side effect of increasing noise. By superimposing several lag-corrected and appropriately shifted images, however, the signal-to-noise ratio can be restored. The algorithm uses the phase-correlation method to measure the two-dimensional shift of a mobile coronary arterial structure. Processing is confined to a rectangular area of interest (AOI), which encloses a feature of clinical significance. The differences of the phases of the Fourier transforms of two frames is computed, combined with an appropriate filter, and inverse Fourier-transformed to produce a phase-correlation image. The vector separation from the origin of image space of the peak of the phase-correlation image is the estimate of the shift of the artery's position in the second frame as compared to the first. The isolation of the AOI from the surrounding image is achieved by the application of a window and correction for any linear trend in the background intensity.     

Migration age profiles for local authority areas in England, 1971-1981

"This paper is a report on continuing research on the application of age profiles in migration analysis. In particular, it is an update of previous work published in this journal with an analysis of 1981 Census data for local authority areas in England. The changes in migration patterns over the intercensal period from 1971 to 1981 are described." The focus is on internal migration     

Use of the quenching factor for predicting the properties of polymer quenching media

The development of methods for evaluating and predicting the variation of the properties of polymer quenching media in operation requires an analysis of the quenching factor, which characterizes the degree of decomposition of the solid solution in quenching, which decreases the strength (hardness) of the alloy by a given value. In the present paper the quenching factorQ is used to compare the properties of aluminum alloys quenched in hot water and aqueous solutions of polymers.Translated from Metallovedenie i Termicheskaya Obrabotka Metallov, No. 6, pp. 14 – 17, June, 1996.