Androgen depletion activates telomerase in the prostate of the nonhuman primate, Macaca mulatta

BACKGROUND: The activity of telomerase, an enzyme that synthesizes telomeric repeats at the ends of chromosomes, is not detectable in normal human prostate. However, the majority of human prostate cancers exhibit telomerase activity. Since androgens play a major role in prostate tumorigenesis, we investigated the effect of androgen-depletion on the expression of telomerase activity in the prostate. METHODS: Adult male rhesus monkeys were either bilaterally castrated or subjected to sham surgery (n = 5 each). Approximately 6 weeks later, the animals were killed and the different regions of the prostate gland were removed and frozen immediately. Telomerase activity was assayed using the telomeric repeat amplification protocol. RESULTS: All five regions of the prostate from sham operated control animals failed to exhibit telomerase activity. In the castrated monkey, all regions of the prostate, except for the anterior lobe, expressed high levels of telomerase activity. CONCLUSIONS: Our results indicate that in monkeys, androgen-ablation leads to up-regulation of telomerase activity. The negative-regulation of telomerase activity by androgens is probably lost during prostate tumorigenesis.     

Anterior corneal optical aberrations induced by photorefractive keratectomy for hyperopia

PURPOSE: Photorefractive keratectomy (PRK) for hyperopia requires both a steepening of the central cornea and a flattening of the mid-periphery to achieve its effect and is likely to affect the optical aberrations of the eye. METHODS: Nine patients underwent PRK to correct between +2.00 and +4.00 D of hyperopia (first eye treated for each patient) using the Summit Technology Apex Plus excimer laser. Anterior corneal aberrations for pupil diameters of 3, 5.5 and 7 mm were estimated from corneal topography data (TMS-1), assuming a uni-index, single surface cornea. Refractive error was assessed using retinoscopy and standard subjective tests. RESULTS: Apart from the intended change in refraction (mean spherical equivalent manifest refraction, +4.60 +/- 1.60 D before surgery and +0.70 +/- 1.60 D at 1 year after surgery), the most significant change was in spherical aberration. Anterior corneal spherical aberration was positive (+1.60 +/- 0.60 D for a 5.5-mm pupil) before surgery and became negative after surgery (-1.80 +/- 1.20 D at 1 year). The change in spherical aberration was related to the achieved change in refractive error. CONCLUSIONS: The large change (approximately 3.00 D) in spherical aberration (from positive to negative aberration) has implications for the optical performance of the whole eye, where the effects of lenticular aberration must also be considered.     

Effect of alpha subunit on allosteric modulation of ion channel function in stably expressed human recombinant gamma-aminobutyric acid(A) receptors determined using (36)Cl ion flux

Inhibitory gamma-aminobutyric acid (GABA)(A) receptors are subject to modulation at a variety of allosteric sites, with pharmacology dependent on receptor subunit combination. The influence of different alpha subunits in combination with beta3gamma2s was examined in stably expressed human recombinant GABA(A) receptors by measuring (36)Cl influx through the ion channel pore. Muscimol and GABA exhibited similar maximal efficacy at each receptor subtype, although muscimol was more potent, with responses blocked by picrotoxin and bicuculline. Receptors containing the alpha3 subunit exhibited slightly lower potency. The comparative pharmacology of a range of benzodiazepine site ligands was examined, revealing a range of intrinsic efficacies at different receptor subtypes. Of the diazepam-sensitive GABA(A) receptors (alpha1, alpha2, alpha3, alpha5), alpha5 showed the most divergence, being discriminated by zolpidem in terms of very low affinity, and CL218,872 and CGS9895 with different efficacies. Benzodiazepine potentiation at alpha3beta3gamma2s with nonselective agonist chlordiazepoxide was greater than at alpha1, alpha2, or alpha5 (P < 0.001). The presence of an alpha4 subunit conferred a unique pharmacological profile. The partial agonist bretazenil was the most efficacious benzodiazepine, despite lower alpha4 affinity, and FG8205 displayed similar efficacy. Most striking were the lack of affinity/efficacy for classical benzodiazepines and the relatively high efficacy of Ro15-1788 (53 +/- 12%), CGS8216 (56 +/- 6%), CGS9895 (65 +/- 6%), and the weak partial inverse agonist Ro15-4513 (87 +/- 5%). Each receptor subtype was modulated by pentobarbital, loreclezole, and 5alpha-pregnan-3alpha-ol-20-one, but the type of alpha subunit influenced the level of potentiation. The maximal pentobarbital response was significantly greater at alpha4beta3gamma2s (226 +/- 10% increase in the EC(20) response to GABA) than any other modulator. The rank order of potentiation for pregnanolone was alpha5 > alpha2 > alpha3 = alpha4 > alpha1, for loreclezole alpha1 = alpha2 = alpha3 > alpha5 > alpha4, and for pentobarbital alpha4 = alpha5 = alpha2 > alpha1 = alpha3.     

Scalp recorded direct current (DC) potential shifts associated with food intake in hungry humans

In humans, eating is assumed to be regulated within a neuronal circuitry integrating hypothalamic "feeding centers" with neocortical regions. Here, DC potentials were recorded in food deprived men to demonstrate a graded tuning of neocortical excitability in conjunction with meal ingestion. In the beginning of food ingestion a pronounced negative DC potential shift developed (P<0.01) which was replaced by a gradual positive potential shift reaching a maximum within 5 min after cessation of food intake (P<0.05). Both negative and positive shifts showed a widespread cortical distribution. The initial negative DC potential presumably reflecting increased depolarisation of apical cortical dendrites, may serve to facilitate eating behavior. The succeeding positivity points to a growing inhibitory influence on cortical processing with increasing satiety that may support termination of meal intake.     

Calcium inhibits willardiine-induced responses in kainate receptor GluR6(Q)/KA-2

A number of studies have demonstrated that willardiine [(S)-1-(2-amino-2-carboxyethyl) pyrimidine-2,4-dione] is a useful agonist for the activation of AMPA/kainate receptors. Here we examine the effect of extracellular calcium on currents evoked by willardiine in HEK 293 cells expressing the GluR6(Q)/KA-2 kainate receptor subunits. At a concentration of 1.8 mM, Ca2+ inhibited the currents induced by 100 microM willardiine by approximately 50%. When extracellular Na+ ions were replaced with Ca2+ ions there were no measurable inward currents. We conclude that Ca2+ inhibition of the willardiine-induced response is concentration dependent.     

Osteoprotegerin is produced when prostaglandin synthesis is inhibited causing osteoclasts to detach from the surface of mouse parietal bone and attach to the endocranial membrane

Osteoclast differentiation and activation is controlled, at least in part, by the counterbalancing influences of osteoprotegerin ligand (OPGL) and osteoprotegerin (OPG). Nonsteroidal anti-inflammatory drugs have been shown to inhibit bone loss in vivo and bone resorption in vitro, and this is associated with a loss of osteoclasts from the bone surface. We test the hypothesis that OPG mediates the inhibition of osteoclast activity that occurs with indomethacin in the mouse calvaria. Recombinant human OPG, like indomethacin, was found to cause osteoclasts to detach from the bone surface and attach to the adjacent endocranial membrane (periosteum). Recombinant human OPG also inhibited the stimulatory effect of prostaglandin E2 (PGE2), parathyroid hormone (PTH), and 1,25-dihydroxyvitamin D3 (1,25D3) on osteoclast adhesion to bone after an incubation with indomethacin. A function-blocking antibody to OPG and soluble human OPGL both inhibited the effect of indomethacin, leaving active osteoclasts on the bone. OPG activity was detected in the culture medium from indomethacin-treated bones and PTH, PGE2, 1,25D3, and dexamethasone all inhibited the production of OPG activity. We conclude that, in the absence of specific stimulators of bone resorption, OPG is produced by the mouse calvaria in vitro, which inhibits bone resorption by causing osteoclasts to detach from the bone surface.     

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 4. The influence of salt speciation, soil type, and sewage sludge on toxicity using the collembolan Folsomia fimetaria and the earthworm Aporrectodea caliginosa as test organisms

Sewage sludge applied to agricultural soils often contains linear alkylbenzene sulfonates (LAS) in the range of 1 to 10 g/kg dry weight, and their toxicity to relevant soil organisms should, therefore, be assessed to ensure safe use of sewage sludge as a fertilizer. Studies of LAS toxicity to soil organisms are few, and to our knowledge, factors that may influence the toxicity in the field have not been studied in detail. In this paper, we report on the influence of speciation of LAS in the test solution added to soil (soluble Na-LAS vs poorly soluble Ca-LAS or Mg-LAS), the influence of soil type, and the modifying effects of sludge amendment on the toxicity of LAS. These issues were investigated using reproduction of Collembola and growth of juvenile earthworms as test parameters. Speciation of the LAS added to test soil did not have any influence on toxicity for any of the test species. Likewise, in three different agricultural soils (sand, loam, and clay), we found almost equal toxicities. The LAS added to test soil in a sludge-water suspension was equally toxic as when it was added in an aqueous solution. However, anaerobic incubation for 7 and 14 d of the LAS-sludge suspension (with no decay of LAS) caused the toxicity to increase almost threefold in both collembolan and earthworm. The relationships between soil constituents, bioavailability, and toxicity are also discussed.     

Bioactive glass coatings with hydroxyapatite and Bioglass particles on Ti-based implants. 1. Processing

Silicate-based glasses with thermal expansion coefficients that match those of Ti6A14V were prepared and used to coat Ti6A14V by a simple enameling technique. Bioglass (BG) or hydroxyapatite (HA) particles were embedded on the coatings in order to enhance their bioactivity. HA particles were immersed partially during heating and remained firmly embedded on the coating after cooling. There was no apparent reaction at the glass/HA interface at the temperatures used in this work (800-840 degrees C). In contrast, BG particles softened and some infiltration into the glass coating took place during heat treatment. In this case, particles with sizes over 45 microm were required, otherwise the particles became hollow due to the infiltration and crystallization of the glass surface. The concentration of the particles on the coating was limited to 20% of surface coverage. Concentrations above this value resulted in cracked coatings due to excessive induced stress. Cracks did not propagate along the interfaces when coatings were subjected to Vickers indentation tests, indicating that the particle/glass and glass/metal interfaces exhibited strong bonds. Enameling, producing excellent glass/metal adhesion with well-attached bioactive particles on the surface, is a promising method of forming reliable and lasting implants which can endure substantial chemical and mechanical stresses.     

Culture of Helicobacter pylori from a gastric string may be an alternative to endoscopic biopsy

Helicobacter pylori was isolated from a swallowed string from 32 of 33 adult subjects (97%) with selective culture media. With this method, antibiotic susceptibility testing and molecular epidemiology studies of H. pylori can be carried out without the need for the collection of specimens by endoscopic biopsy.     

Modulation of rat uterine contractility by mast cells and their mediators

OBJECTIVE: This study was designed to test the possibility that mast cells play a role in the regulation of uterine contractility. STUDY DESIGN: Histamine and rat mast cell protease II levels were determined by radioenzymatic assay and standard radial immunodiffusion techniques, respectively, in uterine tissues from Wistar rats with timed pregnancies. Isolated uterine strips from nonsensitized and ovalbumin-sensitized nonpregnant and pregnant Wistar rats were used for isometric tension recording. Contractile responses to compound 48/80, carbachol, ovalbumin, normal rabbit serum, antirat immunoglobulin E, and 5-hydroxytryptamine were obtained. Antagonists methysergide, ketanserin, 5,8,11, 14-eicosatetraynoic acid, diphenhydramine, and sodium meclofenamate were also used. RESULTS: Tissue levels of rat mast cell protease II and histamine were decreased during delivery compared with prepartum and postpartum levels. Carbachol and compound 48/80 stimulated uterine contractility, and responses were highest during late gestation (day 16 to term). Responses to ovalbumin of uterine tissues in rats sensitized to the antigen were highest at midpregnancy and decreased during the last 10 days of gestation. Ovalbumin challenge in vitro increased the frequency and magnitude of contractions in tissues from ovalbumin-sensitized rats. Compound 48/80 and antirat immunoglobulin E stimulated contractility in both control and sensitized rats. None of the antagonists prevented the contractile responses. CONCLUSIONS: Activation of mast cells is an effective mechanism for stimulation of uterine contractility and may play an important role in the control of term and preterm parturition.