The bestrophin- and TMEM16A-associated Ca2+-activated Cl– channels in vascular smooth muscles

The presence of Ca²⁺-activated Cl^– currents (I_Cl(Ca)) in vascular smooth muscle cells (VSMCs) is well established. I_Cl(Ca) are supposedly important for arterial contraction by linking changes in [Ca²⁺]_i and membrane depolarization. Bestrophins and some members of the TMEM16 protein family were recently associated with I_Cl(Ca). Two distinct I_Cl(Ca) are characterized in VSMCs; the cGMP-dependent I_Cl(Ca) dependent upon bestrophin expression and the ‘classical’ Ca²⁺-activated Cl^– current, which is bestrophin-independent. Interestingly, TMEM16A is essential for both the cGMP-dependent and the classical I_Cl(Ca). Furthermore, TMEM16A has a role in arterial contraction while bestrophins do not. TMEM16A’s role in the contractile response cannot be explained however only by a simple suppression of the depolarization by Cl^– channels. It is suggested that TMEM16A expression modulates voltage-gated Ca²⁺ influx in a voltage-independent manner and recent studies also demonstrate a complex role of TMEM16A in modulating other membrane proteins.     

Protection of Telomeres 1 Is Required for Telomere Integrity in the Moss Physcomitrella patens

In vertebrates, the single-stranded telomeric DNA binding protein Protection of Telomeres 1 (POT1) shields chromosome ends and prevents them from eliciting a DNA damage response. By contrast, Arabidopsis thaliana encodes two divergent full-length POT1 paralogs that do not exhibit telomeric DNA binding in vitro and have evolved to mediate telomerase regulation instead of chromosome end protection. To further investigate the role of POT1 in plants, we established the moss Physcomitrella patens as a new model for telomere biology and a counterpoint to Arabidopsis. The sequence and architecture of the telomere tract is similar in P. patens and Arabidopsis, but P. patens harbors only a single-copy POT1 gene. Unlike At POT1 proteins, Pp POT1 efficiently bound single-stranded telomeric DNA in vitro. Deletion of the P. patens POT1 gene resulted in the rapid onset of severe developmental defects and sterility. Although telomerase activity levels were unperturbed, telomeres were substantially shortened, harbored extended G-overhangs, and engaged in end-to-end fusions. We conclude that the telomere capping function of POT1 is conserved in early diverging land plants but is subsequently lost in Arabidopsis.     

In vivo and in vitro characterization of novel microparticulates based on hyaluronan and chitosan hydroglutamate

This study examined the application of previously characterized microparticles composed of hyaluronan (HA) and chitosan hydroglutamate (CH) as well as novel microparticles consisting of both polymers (HA/CH) to improve the nasal delivery of a model drug. The rabbit bioavailabilities of gentamicin incorporated in HA, CH, and HA/CH microparticles were increased 23-, 31-, and 42-fold, respectively, compared with the control intranasal solution of gentamicin, indicating that all test microparticles were retained for longer periods on the nasal mucosa of the rabbits as supported by previous in vitro dissolution as well as frog palate mucoadhesion studies, thereby improving drug absorption. The higher bioavailabilities of CH-based formulations (CH and HA/CH) suggest the penetration-enhancing effects of CH may also be partially responsible for the improvement. A model was developed, based on a glass impinger device, to deliver dry powder formulations reproducibly onto the surface of cultured cell monolayers. In vitro permeability and fluorescence microscopy studies on the tight junctions of the 16HBE14o- cell lines further confirmed the ability of CH-based formulations to enhance penetration. Furthermore, the in vitro absorption profile from cell culture studies was consistent with those determined from in vivo studies. The complementary effect from the mucoadhesive nature of HA coupled with the penetration-enhancing effects of CH makes the novel HA/CH formulation a promising nasal delivery system.     

One new genus and three new species of Orsillinae (Hemiptera: Lygaeidae) from Australia

Abstract  Two new species of Nysius Dallas, N. orarius sp. n. and N. tasmaniensis sp. n. are described from New South Wales and Tasmania (Australia), respectively. A new monotypic genus, Reticulatonysius , with type-species R. queenslandensis sp. n. is described from Queensland, and its systematic relationship with other orsilline genera is discussed.     

Effects of waterlogging and drought on winter wheat and winter barley grown on a clay and a sandy loam soil

Summary The effects of winter waterlogging and a subsequent drought on the growth of winter barley and winter wheat have been examined. We used lysimeters containing soil monoliths with facilities to control the water table and a mobile shelter to control rainfall. Winter wheat was grown on a clay and on a sandy loam, but winter barley only on the clay soil. Lysimeters were either freely-drained during the winter or waterlogged with the water table 10 cm below the soil surface from 2 December until 31 March (that could occur by rainfall with a return period of 2 to 3 years). The lysimeters then were either irrigated so that the soil moisture deficit did not exceed 84 mm, or subjected to drought by limiting rainfall (equivalent to a 1 in 10 dry year in the driest area of England) so that the deficits reached maximum values of 150 mm in the clay and 159 mm in the sandy loam by harvest.Winter waterlogging restricted tillering and restricted the number of ears for all crops; grain yield of the winter barley was decreased by 219 g/m² (30%), and that of winter wheat by 170 g/m² (24%) and 153 g/m² (21% on the clay and sandy loam respectively.The drought treatment reduced the straw weight of winter barley by 75 g/m² (12%) but did not significantly depress the grain yield. For winter wheat on the clay, where the soil was freely-drained during the winter, drought depressed total shoot weight by 344 g/m² (17%) and grain weight by 137 g/m² (17%), but after winter waterlogging, drought did not further depress total or grain weight. In contrast, the winter wheat on the sandy loam was not significantly affected by drought.From these results, which are discussed in relation to other experiments in the United Kingdom, it seems that winter waterlogging is likely to cause more variation in the yield of winter barley and winter wheat than drought.     

The high-latitude terrestrial carbon sink: a model analysis

A dynamic, global vegetation model, hybrid v4.1 ( Friend et al. 1997 ), was driven by transient climate output from the UK Hadley Centre GCM (HadCM2) with the IS92a scenario of increasing atmospheric CO₂ equivalent, sulphate aerosols and predicted patterns of atmospheric N deposition. Changes in areas of vegetation types and carbon storage in biomass and soils were predicted for areas north of 50°N from 1860 to 2100. Hybrid is a combined biogeochemical, biophysical and biogeographical model of natural, potential ecosystems. The effect of periodic boreal forest fires was assessed by adding a simple stochastic fire model. Hybrid represents plant physiological and soil processes regulating the carbon, water and N cycles and competition between individuals of parameterized generalized plant types. The latter were combined to represent tundra, temperate grassland, temperate/mixed forest and coniferous forest. The model simulated the current areas and estimated carbon stocks in the four vegetation types. It was predicted that land areas above 50°N (about 23% of the vegetated global land area) are currently accumulating about 0.4 PgC y^?1 (about 30% of the estimated global terrestrial sink) and that this sink could grow to 0.8–1.0 PgC y^?1 by the second half of the next century and persist undiminished until 2100. This sink was due mainly to an increase in forest productivity and biomass in response to increasing atmospheric CO₂, temperature and N deposition, and includes an estimate of the effect of boreal forest fire, which was estimated to diminish the sink approximately by the amount of carbon emitted to the atmosphere during fires. Averaged over the region, N deposition contributed about 18% to the sink by the 2080 s. As expected, climate change (temperature, precipitation, solar radiation and saturation pressure deficit) and N deposition without increasing atmospheric CO₂ produced a carbon source. Forest areas expanded both south and north, halving the current tundra area by 2100. This expansion contributed about 30% to the sink by the 2090 s. Tundra areas which were not invaded by forest fluctuated from sink to source. It was concluded that a high latitude carbon sink exists at present and, even assuming little effect of N deposition, no forest expansion and continued boreal forest fires, the sink is likely to persist at its current level for a century.     

Numerical analysis of ryanodine receptor activation by L-type channel activity in the cardiac muscle diad.

Computer simulations were used to examine the response of ryanodine receptors (RyRs) to the sarcolemmal calcium influx via L-type calcium channels (DHPRs). The effects of ryanodine receptor organization, diad geometry, DHPR single-channel current, and DHPR gating were examined. In agreement with experimental findings, the simulations showed that RyRs can respond rapidly (approximately 0.4 ms) to calcium influx via DHPRs. The responsiveness of the RyR depends on the geometrical arrangement between the RyRs and the DHPR in the diad, with wider diads being generally less responsive. When the DHPR single-channel current is small (approximately 25 fA), the organization of RyRs into small clusters results in an improved responsiveness. With experimentally observed DHPR mean open and closed times (0.17 ms and 4 ms, respectively) it is the first opening of the DHPR that is most likely to activate the RyR. A measure of the efficiency (Q) by which DHPR gating evokes sarcoplasmic reticulum release is defined. Q is at maximum for tau approximately 0.3 ms, and we interpret this finding in terms of the "tuning" of DHPR gating to RyR response. If certain cardiac myopathies are associated with a mismatch in the "tuning," then modification of DHPR gating with drugs to "retune" calcium-induced calcium release should be possible.     

Liquid chromatographic analysis of prednisolone, prednisone and their 20-reduced metabolites in perfusion media

A reversed-phase liquid chromatographic assay was developed to quantitate prednisolone, prednisone and the 20α-dihydro and 20β-dihydro reduced metabolites of both parent compounds in tissue culture media from in vitro perfusions of the human placental lobule. Steroids were extracted from perfusate, using reversed-phase cartridges, with average recoveries of 95.2% or greater. The internal standard for the analyses was 6α-methylprednisolone. In this assay cortisol coelutes with prednisolone, however, no other significant interferences were found. Assay of each steroid was linear in the range 0–1 μg/ml. Intra-assay coefficients of variation were measured at 10 and 750 ng/ml with ranges of 3.4% (20α-dihydroprednisone) to 8.8% (20β-dihydroprednisolone) and 4.1% (20β-dihydroprednisone) to 8.8% (prednisone). The corresponding inter-assay coefficients of variation were 3.3% (20α-dihydroprednisone) to 9.1% (20β-dihydroprednisolone) and 1.9% (prednisolone) to 3.5% (prednisone). The analyses utilized two C₁₈ columns which were linked together and maintained at 40°C.     

Effects of intravenous infusion of prostacyclin (PGI2) in man

Prostacyclin infused intravenously in human volunteers induces ex vivo inhibition of platelet aggregation, tachycardia and hypotension. The inhibition of platelet aggregation is obtained with slightly lower doses than those which exhibit cardiovascular effects.The cardiovascular effects disappeared within a few minutes after discontinuing the infusion of prostacyclin but the platelet effects were longer lasting.Prostacyclin did not have any effect on platelet count, platelet factor 3, accelerated partial thromboplastin time, prothrombin time, euglobulin clot lysis time, fibrinogen degradation products, blood glucose concentration or urine sodium potassium ratio.