5-HT3 Receptor-independent Inhibition of the Depolarization-induced 86Rb Efflux from Human Neuroblastoma Cells,TE671, by Ondansetron

The 5-HT₃-receptor antagonist, ondansetron, has been shown to have positive effects in selected in-vivo models of memory impairment and anxiety. The exact mechanisms underlying such bioactivities are unknown. In the present work, an ⁸⁶Rb efflux bioassay was used to show that ondansetron has a unique ability to block voltage-gated potassium channels in TE671 human neuroblastoma cells. This intrinsic potassium-channel-blocking (KCB) property is relatively weak (IC50 20 (M), but is not shared by other 5-HT₃-receptor ligands including zatosetron, MDL 72222, LY 278, 584, zacopride, 1-phenylbiguanide, and ICS 205–930 (tropisetron). Pre-incubation of the target neuroblastoma cells with several 5-HT-receptor ligands including 5-hydroxytryptamine, 8-OH-DPAT, ketanserin, 2-methyl-5-HT, as well as a number of potent 5-HT₃ agonists and antagonists and two selective neurotoxins, failed to abolish the KCB action of ondansetron. A preliminary structure-activity relationship analysis indicates that the KCB activity of ondansetron is almost entirely attributable to its structural nucleus, 2,3-dihyro-9-methyl-4(lH)-carbazolone. It is hypothesized that the KCB action of ondansetron is mediated through receptors other than 5-HT₃ receptors. The KCB activity of ondansetron may be a significant factor in the in-vivo cognition-enhancing activities of this compound, conceivably due to depolarization of the hippocampal synaptic membranes and a consequent augmentation of neurotransmission.     

The Alcohol Beverage Warning Label: When Did Knowledge Increase?

This article presents data on the awareness of the alcohol beverage warning label among a sample of 5,169 inner city African-American gravidas seeking prenatal care. While the label law was implemented in November 1989, a significant increase in knowledge of the label did not occur until March 1990. Women who predominantly consumed wine coolers and beer, and those under age 30 were more likely to know about the label than their counterparts.     

Erythrocyte autoantibodies, autoimmune haemolysis, and carcinoma.

AIMS--To examine a large series of patients in whom both red cell autoantibodies and carcinoma are present; and to determine whether this rare occurrence is a true association or a chance event. METHODS--The laboratory records of 160 patients (76 men, 84 women; mean age 68 years) with erythrocyte autoantibodies and confirmed carcinoma were examined for site of tumour origin and clinical and immunohematological findings. To test whether the concomitant occurrence of autoantibodies and carcinoma was fortuitous, data on total population and carcinoma incidence were included in a chi 2 analysis. RESULTS--The association was significant (chi 2 = 97.5, p < 0.0005); erythrocyte autoantibodies and carcinoma were found together 12-13 times more often than expected from their relative frequencies. Autoantibodies occurred with a variety of carcinomas, particularly those of breast, lung, colon, rectum, and prostate; this largely reflected tumour incidence. Adenocarcinoma, squamous, anaplastic, and transitional cell types were all represented. Warm, cold, and mixed autoantibodies were not associated with particular tumour sites or histology. Eighty six patients had haemolysis of varying severity, 37 had metastatic disease, and 28 died within a few months of presentation. CONCLUSIONS--The presence of erythrocyte autoantibodies and carcinoma in the same patient is a true association and probably reflects a fundamental disturbance in immune homeostasis. It tends to occur with a large tumour mass and metastatic disease, and generally indicates a poor prognosis.     

Gentamicin-induced increases in cytosolic calcium in pig kidney cells (LLC-PK1)

LLC-PK1 cells, an established epithelial cell line derived from pig kidney, were tested as a model system for assessing the role of calcium in gentamicin-induced nephrotoxicity. Cell viability was evaluated by a vital dye exclusion procedure, and intracellular free calcium [Ca2+]i was measured employing Fura-2 fluorescence. Exposing cell suspensions (10(6)/ml) to concentrations of the drug, which had no apparent effect on viability, produced a rapid and prolonged increase in intracellular [Ca2+]. The perturbation of calcium homeostasis could be blocked by the addition of mepiperphenidol, an inhibitor of the organic cation transport system. We propose that LLC-PK1 cells are an appropriate model to study drug-induced nephrotoxicity. Gentamicin disrupts calcium homeostasis and causes plasma membrane alterations. Since mepiperphenidol blocked the gentamicin-induced Ca2+ increases, the data suggest that aminoglycosides enter the cell via the organic cation transporter.     

Competitive control of the self-renewing T cell repertoire

We develop a mathematical model for the self-renewing part of the T cell repertoire. Assuming that self-renewing T cells have to be stimulated by immunogenic MHC-peptide complexes presented on the surfaces of antigen-presenting cells, we derive a model of T cell growth in which competition for MHC-peptide complexes limits T cell clone sizes and regulates the total number of self-renewing T cells in the animal. We show that for a sufficient diversity and/or degree of cross-reactivity, the total T cell number hardly depends upon the diversity of the T cell repertoire or the diversity of the set of presented peptides. Conversely, for repertoires of lower diversity and/or cross-reactivity, steady-state total T cell numbers may be limited by the diversity of the T cells. This provides a possible explanation for the limited repertoire expansion in some, but not all, mouse T cell re-constitution experiments. We suggest that the competitive interactions described by our model underlie the normal T cells numbers observed in transgenic mice, germ-free mice and various knockout mice.      

Localization of a gene for otosclerosis to chromosome 15q25-q26

Among white adults otosclerosis is the single most common cause of hearing impairment. Although the genetics of this disease are controversial, the majority of studies indicate autosomal dominant inheritance with reduced penetrance. We studied a large multi- generational family in which otosclerosis has been inherited in an autosomal dominant pattern. Five of16 affected persons have surgically confirmed otosclerosis; the remaining nine have a conductive hearing loss but have not undergone corrective surgery. To locate the disease- causing gene we completed genetic linkage analysis using short tandem repeat polymorphisms (STRPs) distributed over the entire genome. Multipoint linkage analysis showed that only one genomic region, on chromosome 15q, generated a lod score >2.0. Additional STRPs were typed in this area, resulting in a lod score of 3.4. STRPs FES (centromeric) and D15S657 (telomeric) flank the 14. 5 cM region that contains an otosclerosis gene.      

IMPT1, an imprinted gene similar to polyspecific transporter and multi- drug resistance genes

Human chromosome 11p15.5 and distal mouse chromosome 7 include a megabase-scale chromosomal domain with multiple genes subject to parental imprinting. Here we describe mouse and human versions of a novel imprinted gene, IMPT1 , which lies between IPL and p57 KIP2 and which encodes a predicted multi-membrane-spanning protein similar to bacterial and eukaryotic polyspecific metabolite transporter and multi- drug resistance pumps. Mouse Impt1 and human IMPT1 mRNAs are highly expressed in tissues with metabolite transport functions, including liver, kidney, intestine, extra-embryonic membranes and placenta, and there is strongly preferential expression of the maternal allele in various mouse tissues at fetal stages. In post-natal tissues there is persistent expression, but the allelic bias attenuates. An allelic expression bias is also observed in human fetal and post-natal tissues, but there is significant interindividual variation and rare somatic allele switching. The fact that Impt1 is relatively repressed on the paternal allele, together with data from other imprinted genes, allows a statistical conclusion that the primary effect of human chromosome 11p15.5/mouse distal chromosome 7 imprinting is domain-wide relative repression of genes on the paternal homolog. Dosage regulation of the metabolite transporter gene(s) by imprinting might regulate placental and fetal growth.      

Characterization of chloroquine-induced autophagic vacuoles isolated from rat liver

Studies have been undertaken to investigate the role of cellular autophagy in the accommodation of stress in a biological system. Chloroquine (Aralen hydrochloride), an antimalarial and anti-inflammatory drug, was used to induce autophagy in rat liver. A method is presented which uses differential and discontinuous sucrose gradient centrifugation for the preparation of autophagic vacuole-enriched fractions from rat liver. Ultrastructural studies of the autophagic vacuole fractions showed that the integrity of the autophagic vacuoles was maintained throughout the isolation procedure and that they were morphologically similar to those seen in situ. Assay of glucose-6-phosphatase, NADPH-DCIP reductase, and acid phosphatase confirm the presence of membranes derived from the endoplasmic reticulum, as well as lysosomes, in the autophagic vacuole fractions. The distribution of [¹⁴C]-chloroquine suggested a preferential binding of the drug to the autophagic vacuoles may have occurred. These results suggest that cellular autophagy may play an important role in the accommodation of chemically induced alterations in hepatocytes by preferentially sequestering chloroquine, as well as restoring cellular ultrastructure.