Haemopoietic growth factor production by normal and aplastic anaemia stroma in long-term bone marrow culture

Defective marrow stroma, or microenvironment, have been proposed as one of several mechanisms to account for bone marrow failure in aplastic anaemia (AA). This could involve defects in positive- or negative-acting haemopoietic regulator expression by AA stroma, or alteration of normal stroma-stem cell interactions. We have used a sensitive bioassay to investigate production of granulocyte-colony stimulating factor (G-CSF), granulocyte-macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-3, IL-6 and stem cell growth factor (SCF), by normal and AA stroma in long-term bone marrow culture (LTBMC). LTBMC were grown to confluence, irradiated and harvested to yield a single cell suspension. These cells were cocultured with normal target bone marrow mononuclear cells (BMMC), or CD34+ cells, in clonogenic assays, in the absence of exogenous cytokines. Cytokines responsible for the colony-stimulating activity (CSA) and burst-promoting activity (BPA) produced by stromal cells were identified by neutralizing antibodies to specific cytokines. All normal stroma populations produced G-CSF and GM-CSF, 93% produced IL-3, 80% produced IL-6, and 70% produced SCF. Similarly, all AA stroma produced G-CSF and GM-CSF, and 71% produced SCF. In contrast, only 71% of AA stroma produced IL-3 and 36% produced IL-6. Target cell stimulation was not dependent on direct stroma-target cell contact, suggesting production of soluble cytokines. However, although both IL-6 and G-CSF were detected in LTBMC supernatants by enzyme-linked immunoassay (ELISA), IL-3 and GM-CSF were undetectable, perhaps indicating low-level local production of these factors.     

GLC analysis of thioridazine, mesoridazine, and their metabolites

A GLC method for measuring thioridazine, mesoridazine, their metabolites, and possibly other phenothiazines was developed. By using this method, seven different phenothiazine derivatives, thioridazine, and six known thioridazine metabolites were extracted and separated. This method was tested by assaying plasma samples from 30 hospitalized psychiatric patients receiving thioridazine or mesoridazine.     

Effect of indole-3-acetic acid on the kinetics of horseradish peroxidase catalyzed scopoletin oxidation

The kinetics of horseradish peroxidase catalyzed of scopoletin oxidation were observed to be sigmoidal. The apparent KmS for scopoletin is 0.9 mM. Inhibition of scopoletin oxidation by indole-3-acetic acid (IAA) appears to be non competitive. Non competitive inhibition by IAA suggests a more significant role of the peroxidase protein matrix in regulating the activity of the heme moiety.     

Proton-induced x-ray emission analysis of single human hair roots

Collimated beams of 3.75 million electron volt protons were used to examine a 2-millimeter length of the root end of human hair; the concentrations of some hair root elements were correlated with the results of standard clinical assays of blood samples. The technique should be useful for the analysis of micro amounts of biological tissue.     

Absence of opioid stress-induced analgesia in mice lacking beta-endorphin by site-directed mutagenesis

A physiological role for beta-endorphin in endogenous pain inhibition was investigated by targeted mutagenesis of the proopiomelanocortin gene in mouse embryonic stem cells. The tyrosine codon at position 179 of the proopiomelanocortin gene was converted to a premature translational stop codon. The resulting transgenic mice display no overt developmental or behavioral alterations and have a normally functioning hypothalamic-pituitary-adrenal axis. Homozygous transgenic mice with a selective deficiency of beta-endorphin exhibit normal analgesia in response to morphine, indicating the presence of functional mu-opiate receptors. However, these mice lack the opioid (naloxone reversible) analgesia induced by mild swim stress. Mutant mice also display significantly greater nonopioid analgesia in response to cold water swim stress compared with controls and display paradoxical naloxone-induced analgesia. These changes may reflect compensatory upregulation of alternative pain inhibitory mechanisms.     

Applications of asymmetric collimation on linear accelerators

Frequency of use of asymmetric collimation (AC) at an academic radiation oncology center equipped with AC-capable linear accelerators was determined, and the type of use was cataloged. Records of patients beginning radiation treatment at U.C. Davis Cancer Center within a 3-month period (3/1/92 to 5/31/92) were reviewed. Forty-seven percent of 102 patients and 56% of 123 courses of treatment involved AC. Six common uses of AC were identified: beam-split field matching, planned boosts, other field size changes, adjustments to match divergent fields, matchline feathering, and opposed tangential fields. This study demonstrates that asymmetric collimation is a useful and powerful clinical treatment tool with widespread applications to radiation therapy.     

Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae at a resolution of 2.6 kilobase pairs

Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae are presented. In order of increasing size, they are chromosomes I, VI, III, IX, V and VIII, comprising 2.49 megabase pairs of DNA. The maps are based on the analysis of an overlapping set of lambda and cosmid clones. Overlaps between adjacent clones were recognized by shared restriction fragments produced by the combined action of EcoRI and HindIII. The average spacing between mapped cleavage sites is 2.6 kb. Five of the six chromosomes were mapped from end to end without discontinuities; a single internal gap remains in the map of chromosome IX. The reported maps span an estimated 97% of the DNA on the six chromosomes; nearly all the missing segments are telomeric. The maps are fully cross-correlated with the previously published SfiI/NotI map of the yeast genome by A. J. Link and M. V. Olson. They have also been cross-correlated with the yeast genetic map at 51 loci.