Improved DNA markers for efficient analysis of fragile X families

We report the characteristics of two new probes that detect BclI RFLPs useful for analysis of fragile X families. With these two probes and a single blot, 34% of women are heterozygous both for the proximal marker DXS105 (closer to the fragile X locus than the factor IX gene) and for the distal markers DXS52 or the factor VIII gene. Combined with the analysis of previously described polymorphic markers, it is possible to have a majority of families fully informative for flanking markers using a limited number of probes and restriction digests.     

Multiple tachykinin pools in sensory nerve fibres in the rabbit iris

A population of sensory nerve fibres in the rabbit iris is known to contain calcitonin gene-related peptide and tachykinins, such as substance P and neurokinin A. In the presence of atropine and guanethidine, the isolated iris sphincter responded to electrical stimulation with a contraction that could be abolished by tachykinin antagonists. Capsaicin, known to release tachykinins from sensory fibres, evoked a long-lasting tachykinin-mediated contraction of the iris sphincter. Repeated application of capsaicin led to tachyphylaxis, possibly reflecting depletion of releasable neuronal stores of tachykinins. At this stage, electrical stimulation failed to elicit contraction. The capacity of capsaicin to release neuropeptides from sensory fibres was confirmed by determination of substance P- and calcitonin gene-related peptide-like immunoreactivity in the incubation medium and in the iris tissue. The concentrations of substance P and calcitonin gene-related peptide in the iris after capsaicin exposure were reduced by about 25%. Like capsaicin, bradykinin evoked a tachykinin-mediated contraction and tachyphylaxis. However, after development of tachyphylaxis to bradykinin, electrical stimulation or exposure to capsaicin still evoked tachykinin-mediated contraction, albeit a reduced one compared with the response before bradykinin. Hence, capsaicin completely depletes tachykinin stores releasable by prolonged electrical stimulation, whereas bradykinin exhausts only a sequestered pool. The possibility that tachykinins occur in several releasable pools in sensory nerves was investigated in yet another way: the iris sphincter muscle was stimulated electrically once every 2.5 min over several hours. The contractile response diminished gradually.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of centrally administered neuropeptide Y (NPY) and NPY13–36 on the brain monoaminergic systems of the rat

Summary The effects of centrally administered NPY on the brain monoamine systems were investigated in the rat. Neuropeptide Y (0.2–5.0 nmol), its C-terminal 13–36 amino acid (a.a.) fragment, NPY_13–36 (0.4–10.0 nmol), or saline were injected into the right lateral cerebral ventricle of unrestrained rats. After l h the animals were decapitated, and the brains were taken out. Two cortical regions (frontal and parietal), the striatum, the hypothalamus, and the brain stem were dissected out. The tissue contents of noradrenaline (NA), dopamine (DA) and serotonin (5-HT), as well as of their major metabolites, 3-methoxy-4-hydroxy-phenylethylené glycol (MHPG), 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxy-indole acetic acid (5-HIAA) were measured. The most consistent finding was a dose-related increase of both DA and DOPAC levels after treatment with NPY. This effect was reproduced by NPY_13–36 in cortical tissue, whereas, in the sub-cortical regions, NPY_13–36 only reproduced the effects of NPY on the DOPAC levels. Less consistent effects were found on the NA systems, in which NA levels showed a tendency to increase following low, and decrease after high doses of NPY. These effects were largely reproduced by NPY_13–36. In addition, NPY increased tissue levels of MHPG in frontal cortical tissue in a dose-related manner. The brain 5-HT systems were not affected.     

Intraoperative retinal light damage reflected in electrophysiologic data

In a series of 30 unilaterally pseudophakic patients, electroretinograms and electrooculograms were recorded 6 months postoperatively. The unoperated on fellow eyes served as controls High intraoperative retinal light exposure (3.4–7.3 mW/cm², Zeiss OPMI 6 operating microscope) caused a substantial reduction of electrophysiologic potentials. Light protection prevented deterioration of electroretinogram and electro-oculogram potentials; reducing the bulb voltage, tilting the axis of illumination, filtering short wavelengths and the use of light shields resulted in 4-log-unit lower intensities (0.8–3.7 W/cm²).Abbreviations ACL anterior chamber lens - ECCE extracapsular cataract extraction - ICCE intracapsular cataract extraction - PCL posterior chamber lens     

Expression of fibrinogen receptors during activation and subsequent desensitization of human platelets by epinephrine

Epinephrine causes platelet aggregation and secretion by interacting with alpha 2-adrenergic receptors on the platelet surface. Platelet aggregation requires the binding of fibrinogen to a specific receptor on the membrane glycoprotein IIb-IIIa complex. Although the IIb-IIIa complex is identifiable on the surface of resting platelets, the fibrinogen receptor is expressed only after platelet activation. The current studies were designed to examine the effect of occupancy of platelet alpha 2-adrenergic receptors by epinephrine on the expression of fibrinogen receptors and on the aggregation of platelets. The ability of epinephrine to induce the expression of fibrinogen receptors was studied under two different conditions: acute stimulation (less than 1 min) and prolonged stimulation (50 to 90 min), the latter of which is associated with a reduction or "desensitization" of the platelet aggregation response. Expression of the fibrinogen receptor was monitored with 125I-fibrinogen as well as with 125I-PAC-1 (PAC-1), a monoclonal antibody that binds to the glycoprotein IIb-IIIa complex only after platelets are activated. Epinephrine caused an immediate increase in PAC-1 and fibrinogen binding that was dependent on occupancy of the alpha 2-receptor by epinephrine and on the presence of extracellular free Ca (KCa = 30 mumol/L). By itself, 1 mmol/L Mg was unable to support induction of the fibrinogen receptor by epinephrine. However, it did decrease the Ca requirement by about two orders of magnitude. Prolonged stimulation of unstirred platelets by epinephrine led to a 70% decrease in the aggregation response when the platelets were subsequently stirred. Despite their decreased aggregation response, desensitized platelets bound PAC-1 and fibrinogen normally, indicating that the loss of aggregation was not due simply to a decrease in fibrinogen receptor expression. Although desensitization was not affected by pretreatment of the platelets with aspirin, it was partially prevented when extracellular Ca was chelated by EDTA during the long incubation with epinephrine. These studies demonstrate that once platelet alpha 2-adrenergic receptors are occupied by epinephrine, extracellular Ca is involved in initiating the aggregation response by supporting the induction of the fibrinogen receptor and the binding of fibrinogen. Furthermore. Ca-dependent reactions subsequent to fibrinogen binding may be necessary for maximal platelet aggregation and are impaired when platelets become desensitized to epinephrine.     

Chlorambucil therapy in hairy cell leukemia: effects on lipid composition and lymphocyte subpopulations

Two patients with progressive hairy cell leukemia following splenectomy were treated with low-dose daily chlorambucil. Both had an objective hematologic response as determined by a return to normal hematocrit and platelet count. This was also reflected in the mononuclear cell fraction by the normalization of cholesterol content, cholesterol/phospholipid ratio, and the lymphocyte subpopulations. This article confirms previous reports on the efficacy of chlorambucil in this setting and describes some morphological, and biochemical concomitant events.