Cytosol Mg2+ modulates Ca2+ ionophore induced secretion from rabbit neutrophils

AbstractThe influence of extracellular Mg2+ on Ca2+ ionophore (A23187 and ionomycin) induced secretion and changes in the cytosol pH of rabbit neutrophils suspended in Ca2+-free buffer has been investigated. While extracellular Ca2+ is obligatory for ionomycin induced secretion, we have defined conditions under which A23187 can induce secretion in Ca2+-free media. The different behaviour of these two Ca2+ ionophores is discussed on the basis of their different counter cation specificities

Chelation of cytoplasmic Ca2+ increases plasma membrane permeability in murine macrophages.

Cytoplasmic free Ca2+ (Ca2+i) was chelated to 10-20 nM in the macrophage cell line J774 either by incubation with quin2 acetoxymethyl ester in the absence of external Ca2+ (Di Virgilio, F., Lew, P.D., and Pozzan, T. (1984) Nature 310, 691-693) or by loading [ethyl-enebis(oxyethylenenitrilo)]tetraacetic acid (EGTA) into the cytoplasm via reversible permeabilization of the plasma membrane with extracellular ATP (Steinberg, T.H., Newman, A.S., Swanson, J.A., and Silverstein, SS.C. (1987) J. Biol. Chem. 262, 8884-8888; Di Virgilio, F., Meyer, B.C., Greenberg, S., and Silverstein, S.C. (1988) J. Cell Biol. 106, 657-666). After removal of ATP from the incubation medium, ATP-permeabilized Ca2+i-depleted macrophages recovered a near-normal plasma membrane potential which slowly depolarized over a 2-4 h incubation at low [Ca2+]i. In both ATP-treated and quin2-loaded cells, depolarization of plasma membrane potential was paralleled by an increase in plasma membrane permeability to low molecular weight aqueous solutes such as eosin yellowish (Mr 692), ethidium bromide (Mr 394), and lucifer yellow (Mr 463). This increased plasma membrane permeability was not accompanied by release of the cytoplasmic marker lactic dehydrogenase for incubations up to 4 h and was likely a specific effect of Ca2+i depletion since it was not caused by: (i) the mere incubation of macrophages with extracellular EGTA, i.e. at near-normal [Ca2+]i; and (ii) loading into the cytoplasm of diethylenetriaminepentaacetic acid, a specific chelator of heavy metals with low affinity for Ca2+. Treatment of Ca2+i-depleted cells with direct (phorbol 12-myristate 13-acetate) or indirect (platelet-activating factor) activators of protein kinase C prevented the increase in plasma membrane permeability. Down-regulation of protein kinase C rendered Ca2+i-depleted macrophages refractory to the protective effect of phorbol 12-myristate 13-acetate. This report suggests a role for Ca2+i and possibly protein kinase C in the regulation of plasma membrane permeability to low molecular weight aqueous solutes

A 1.82 m^2 ring laser gyroscope for nano-rotational motion sensing

We present a fully active-controlled He-Ne ring laser gyroscope, operating in square cavity 1.35 m in side. The apparatus is designed to provide a very low mechanical and thermal drift of the ring cavity geometry and is conceived to be operative in two different orientations of the laser plane, in order to detect rotations around the vertical or the horizontal direction. Since June 2010 the system is active inside the Virgo interferometer central area with the aim of performing high sensitivity measurements of environmental rotational noise. So far, continuous not attempted operation of the gyroscope has been longer than 30 days. The main characteristics of the laser, the active remote-controlled stabilization systems and the data acquisition techniques are presented. An off-line data processing, supported by a simple model of the sensor, is shown to improve the effective long term stability. A rotational sensitivity at the level of ten nanoradiants per squareroot of Hz below 1 Hz, very close to the required specification for the improvement of the Virgo suspension control system, is demonstrated for the configuration where the laser plane is horizontal

Rotational sensitivity of the "G-Pisa" gyrolaser

G-Pisa is an experiment investigating the possibility to operate a high sensitivity laser gyroscope with area less than $1 \rm m^2$ for improving the performances of the mirrors suspensions of the gravitational wave antenna Virgo. The experimental set-up consists in a He-Ne ring laser with a 4 mirrors square cavity. The laser is pumped by an RF discharge where the RF oscillator includes the laser plasma in order to reach a better stability. The contrast of the Sagnac fringes is typically above 50% and a stable regime has been reached with the laser operating both single mode or multimode. The effect of hydrogen contamination on the laser was also checked. A low-frequency sensitivity, below $1 \rm Hz$, in the range of $10^{-8} \rm {(rad / s)/ \sqrt{Hz}}$ has been measured.Comment: 6 pages, 6 figures, presented at the EFTF-IFCS joint conference 200

Voltage-dependent activation and inactivation of calcium channels in PC12 cells. Correlation with neurotransmitter release.

The existence and mechanisms of inactivation of voltage-gated Ca2+ channels are important, but still debatable, physiological problems. By using the Ca2+ indicators quin2 and fura-2, we demonstrate that in PC12 cells voltage-gated Ca2+ channels undergo inactivation dependent on both voltage and [Ca2+]i. Inactivation, however, is never complete and a small number of channels remains open during prolonged depolarization, explaining the steady state elevation of [Ca2+]i observed in cells depolarized with high KCl. A close parallel exists between Ca2+ channel inactivation and the transient nature of neurotransmitter release: secretion is rapidly stimulated during the first 30 s of depolarization, when a transient overshoot in [Ca2+]i can be demonstrated, while it is negligible during the following period, despite the persistence of an elevated [Ca2+]i; predepolarization in Ca2+-free medium and subsequent addition of Ca2+ (a condition which allows the development of the voltage inactivation) abolishes the fast phase of secretion, while not modifying the steady state [Ca2+]i eventually attained; and increases in the intracellular Ca2+ buffering decreases the amplitude of the fast secretion phase induced by KCl without altering the steady state [Ca2+]i. We suggest that localized [Ca2+]i gradients form close to the plasma membrane shortly after depolarization and that the [Ca2+]i reached in these regions is the relevant parameter in the regulation of secretion

Increased sensitivity of DMD lymphoblastoid cell to low doses of X-irradiation.

Several cell membrane abnormalities affecting various cell populations have been reported in Duchenne muscular dystrophy (DMD) by different investigators. In peripheral blood lymphocytes intrinsic cellular membrane defect evidentiated by impairment of capping capacities has been repeatedly obtained, suggesting that DMD product could act in such cellular phenotype at the cytoskeletal compartment. It has been previously reported that lymphoid cells are characterized by high radiosensitivity. On the assumption that DMD phenotypes could increase this susceptibility, we have compared the radiosensitivity of normal and DMD lymphoblastoid cell lines (LCLs) to small doses (0-2Gy) of x-irradiation. The results obtained suggest an increased sensitivity of DMD cells without Ca++ uptake or apoptotic phenomena, associated with an effect upon cell cycle length

Cost-Effectiveness of Vaccination with the 20-Valent Pneumococcal Conjugate Vaccine in the Italian Adult Population

The availability of a new 20-valent pneumococcal conjugate vaccine (PCV) makes it appropriate to assess its cost-effectiveness. This was evaluated by adopting the Italian National Health Service perspective, using a cost consequences Markovian model. The expected effects of vaccination with 20-valent PCV were compared with the administration of 13-valent PCV and 15-valent PCV. Assuming a 100% vaccination of cohorts aged 65–74 years, in the (lifetime) comparison between 20-valent PCV and 13-valent PCV, the former is dominant (lower cost for a better health outcome). A reduction in disease events was estimated: −1208 deaths; −1171 cases of bacteraemia; −227 of meningitis; −9845 hospitalised all-cause nonbacteremic pneumonia cases (NBP) and −21,058 non-hospitalised. Overall, in the Italian population, a total gain of 6581.6 life years and of 4734.0 QALY was estimated. On the cost side, against an increase in vaccinations costs (EUR +40.568 million), other direct health costs are reduced by EUR 48.032 million, with a net saving of EUR +7.464 million. The comparison between 20-valent PCV and 15-valent PCV results in an Incremental Cost-Effectiveness Ratio (ICER) of EUR 66 per life year gained and EUR 91 per QALY gained. The sensitivity analyses confirm the robustness of the results. We can conclude that the switch to 20-valent PCV is a sustainable and efficient investment

Tumor promoter phorbol myristate acetate inhibits Ca2+ influx through voltage-gated Ca2+ channels in two secretory cell lines, PC12 and RINm5F.

Protein kinase C is known to be involved both in initiation and termination of cellular responses due to phosphoinositide breakdown. Here we report that in PC12 cells (a line of neurosecretory cells derived from a rat pheochromocytoma), pretreatment with nanomolar concentrations of phorbol myristate acetate, PMA, which is believed to specifically activate protein kinase C, inhibits the cytosolic-free Ca2+ concentration rise induced by depolarizing agents. In contrast, plasma membrane potential and 45Ca efflux from preloaded cells were unaffected by PMA pretreatment. Inhibition by PMA and diacylglycerol of the cytosolic-free Ca2+ concentration rise induced by depolarization was observed also in another cell line, the insulin secreting line RINm5F. These results raise the possibility that the voltage-gated Ca2+ channel is under inhibitory control by protein kinase C