The size and morphology Etoposide ic50 of synthesized GNPs were recorded using transmission electron microscope, TEM (JEM-100CX.TEM JEOL, Japan). TEM studies were carried by drop coating GNPs onto carbon-coated TEM grids. The film on the TEM grids were allowed to dry, the extra solution was removed using blotting paper. The effects of temperature (40°–100° with 10 °C increments), radiation (2, 2.5, 3, 3.5, 4, 5 and 6 kGy) and different concentrations of tetrachloroauric acid (0.1, 0.2, 0.3, 0.4 and 0.5 ml of 10 mg/ml solution) were assessed to study their effect on the formation of GNPs. Blank sample was done in case of radiation before mixing with HAuCl4. After screening of the seven fungal strains,

Pleurotus ostreatus was chosen due to its relatively high laccase activity (4610 U/gfs), (where gfs is defined as the number of units of enzyme produced from 5 g fermented substrate), compared to other fungi and

consequently, Pleurotus ostreatus was used to optimize laccase production throughout the whole study. The screening of the four agricultural wastes showed that wheat bran which is an abundant byproduct formed during wheat flour preparation was the highest in laccase activity 4610 (U/gfs) compared to other sources (Table 1). When screening different nitrogen sources, malt extract gave the highest laccase activity (8460 U/gfs) compared to yeast extract (7205 U/gfs) and tryptone (6500 U/gfs) (Table 2). Wheat bran and malt extract were both used to support growth of Pleurotus ostreatus. After carrying out 32 experiments, reflecting different combinations of the variables (Table shown in supplementary PLX-4720 in vitro data), the results revealed that run number 28 gave the highest enzyme activity of (32,450 U/gfs) with all of the variables in their higher level except for tannic acid

that is in its lower level. Main effects Cepharanthine plotting showed the positive effect of nitrogen source, Tween-80, CuSO4 and methionine. In case of nitrogen source effect, a major difference between means was noticed. However, tannic acid had negative effect in its higher level as shown in Fig. 1. The significance of the effect of the six variables was assessed using one way (ANOVA). Both the T-value and P-value statistical parameters were used to confirm the significance of factors studied as shown in Table 3. The results shown proved that nitrogen source, Tween-80 and tannic acid all had a significant effect with (P = <0.05). The model determination coefficient (R = 0.81) suggested that the fitted model could explain 81% of the total variation which implies a satisfactory representation of the process by the model. The coeffecient of determination (R-value) always lies between 0 and 1. As the closer the value of R is to 1.0, the stronger the model and the better it predicts the response. The analysis of variance for the selected factorial model showed that the model was significant with a model F-value of 17.75 (P = <0.

In Raja Ampat, published and unpublished information and expert opinion on oceanography, bathymetry and physico-chemical

parameters, habitats and distributions of coral communities and reef fishes were used to develop a more detailed reef classification comprising 14 broad scale reef types termed ‘reefscapes’ (scale of 100–1000s km) and 75 reef habitats click here (scale of 10–100s km) (Fig. 7, Supplementary materials, DeVantier et al., 2009). Reef endemism is high, with 5–6% of all coral species and 2.5% of reef fish found only in this region (Allen and Erdmann, 2012). Unlike many other parts of Indonesia and wider Southeast Asia (Burke et al., 2011), the coral reefs in the BHS are in a relatively healthy state. Reef health monitoring in 9 of the 12 BHS MPAs using point intercept transect methods (Wilson and Green, 2009) showed average live hard coral cover ranged from 14.3% to 44.4% across all the MPAs (Table 1). Manta towing (English et al., 1997) covering a much wider area of the MPAs recorded average coral cover ranging from 15.2% AZD6244 order to 33.6 across all the MPAs (Table 1). The main threats to coral reefs are from destructive fishing such as bomb, cyanide and compressor fishing, though this does not occur to the same intensity or geographic spread as other parts of Indonesia, and is

mainly done by outside fishers frequenting the area (McKenna et al., 2002 and Ainsworth et al., 2008). There is no documentation of major widespread crown-of-thorns starfish (Acanthaster planci) outbreaks on reefs in the BHS. Damaged reefs in the BHS MPAs (based on percentage of rubble), ranged from 11.8% to 24.0% and 8.8% to 33.4% in point intercept transect and manta towing surveys, respectively ( Table 1). Sulfite dehydrogenase Formal patrols with enforcement agencies and informal patrols with local communities have been largely effective in reducing and in some case stopping destructive fishing in MPAs (TNC and CI, unpublished data). However, overfishing continues and is largely uncontrolled (see Section 5.2 for details) and poses a significant and growing threat to coral reefs. Marine lakes are land-locked water bodies that have a marine character maintained

by tidal fluctuations pushing seawater through subterranean crevices or porous karst (Becking et al., 2009). At least 45 marine lakes have been identified in Raja Ampat, with the highest numbers occurring in Kawe and Southeast Misool MPAs (Becking et al., 2009 and Becking et al., 2011). These lakes vary in biophysical parameters such as bathymetry, size, coastline, salinity, water temperature, pH and degree of connection to the sea, which results in a variety of biotic assemblages (Fig. 8; Becking et al., 2011). Fauna observed in Raja Ampat’s lakes include corals, nudibranchs, shrimps, fish, bivalves, sponges (including a number of endemic species), ascidians, ctenophores, and jellyfish including Cassiopeia, Mastigias and Aurelia spp. ( Becking et al., 2009).

In animals submitted to the therapy with MK0431 (group II) it was observed involuted serous acini. However, a recovery was noted when compared to untreated animals (Fig. 1A and Table 3 and Table 4). Stromal spaces filled with extracellular matrix were identified between acini by Picrosirius red staining. The quantity of collagen fibres was significantly selleck kinase inhibitor minor than that

observed in untreated animals (Fig. 1C and Table 5). Pleomorphic serous acini characterized by a reduced spatial area occupied by secretory epithelium were observed in parotid glands of the group I (Fig. 1B and Table 3 and Table 4). The stroma was found enlarged, with a higher volume density of collagen fibres (Fig. 1D and Table 5). Similarly, animals submitted to therapy with MK0431 (group II) presented also involuted seromucous acini; however, Endocrinology antagonist a significant recovery was also noted when compared to untreated animals (Fig. 2A and Table 3 and Table 4). In the same way, stromal spaces filled with extracellular matrix were identified between acini by Picrosirius red staining. The quantity of collagen fibres was significantly minor (Fig. 2C and Table 5).

In submandibular glands, atypical and involuted seromucous acini were observed in the group I (Fig. 2B and Table 3 and Table 4). Enlargement of the interacinar spaces were also observed. Extracellular matrix alterations were observed Celecoxib in the stroma, with the observation of increase in the connective tissue component (Fig. 2D and Table 5). Diabetic animals presented the lowest weight throughout the experimental period. Diabetes mellitus causes metabolic disorders and body weight alteration.29, 30, 31 and 32 Animals submitted to glycaemic treatment, showed recovery of body weight.33

Body weight recovery and gain were observed also after use of incretin-based therapies; however, this cannot reflect an adequate metabolic control. An alternative to the diabetes treatment and weight control is the use of DDPIV inhibitors, as the sitagliptin (MK0431). This incretin mimetic promotes the maintenance and in different cases the loss of weight, as observed in both type 1 and 2 diabetes.14, 17, 18, 34, 35 and 36 Therefore, while weight gain can exacerbate hyperglycemia, the minor weight observed in treated animals may be related to the beneficial effect of treatment with this DPPIV inhibitor. As per to glucose levels, it was observed elevated levels throughout the experimental period in animals of group I, and a significant reduction of glucose levels was observed in animals of group II. In a study using insulin replacement therapy, a proven hypoglycaemic treatment, Hu et al. 37 showed that normal glucose levels in healthy animals are close to 180 mg/dL, whereas mean levels of 300 mg/dL or higher indicate an effective diabetic state.

8 Despite of the numerous studies about the presence of podoplanin expression in various oral tissues and tumours, little is known about its physiologic or pathologic function. Sawa et al.15 suggested an association of podoplanin in cellular proliferative activity due to its expression in tooth germ, which is present in cells with high mitotic activity, i.e. in dental lamina, terminal portion of Hertwig sheath and pre-ameloblasts. Tsuneki et al. 13 www.selleckchem.com/Bcl-2.html found that podoplanin-positive cells are located within areas with PCNA-positive cells in ameloblastomas, keratocystic odontogenic tumours, adenomatoid odontogenic tumours, and calcifying cystic odontogenic tumours. 13 On the other hand, a previous study conducted

by our research group has showed absence of significant correlation between podoplanin and epithelial odontogenic proliferative activity in ameloblastomas reinforcing that the exact role of this protein http://www.selleckchem.com/products/Trichostatin-A.html in the benign odontogenic tumours needs to be elucidated. 14 In view of the above considerations, the aim of this study was to investigate the expression of podoplanin in two groups of odontogenic tumours: those exclusively composed by epithelial neoplastic components and

those composed by epithelial and ectomesenchymal tumoral cells. Additionally, we verified the possible association between podoplanin immunoexpression and the proliferative activity in keratocystic odontogenic tumours and orthokeratinized odontogenic cysts. Fifty-four odontogenic tumours were selected from the archives of the Laboratory of Pathology, Bauru School of Dentistry

– University of São Paulo, Brazil, for the current study: Odontogenic epithelium without ectomesenchyme: • 8 ameloblastomas (AM): 4 follicular and 4 plexiform subtypes; Odontogenic epithelium with ectomesencyhme: • 2 ameloblastic fibromas (AF); The tumours were stained with haematoxylin–eosin and reviewed according to the World Health Organization histological Ergoloid classification of odontogenic tumours.16 This study was approved by the Research Ethics Committee of the Bauru School of Dentistry, University of São Paulo (process number 99/2010). A formalin-fixed 4-μm section of epithelial odontogenic tumours was taken from the pathology archive for immunohistochemistry analysis of anti-podoplanin and anti-Ki-67 antibodies expressions by odontogenic cells. Only KCOTS and OOC were submitted to the Ki-67 antibody reaction. After antigen retrieval using 10 mM citrate buffer, pH 6.0, in a domestic pressure cooker (Nigro, model Eterna 4(1/2) L, Brazil) for 4 min, endogenous peroxidase activity was blocked by incubation in 3% H2O2 for 20 min. Each epithelial odontogenic tumour section was incubated overnight at 4 °C with the primary monoclonal anti-podoplanin antibody (D2-40 clone, code#3619-1; Dako North America, Inc., Carpinteria, CA, USA), dilution 1:200 or anti-Ki-67 antibody (MIB-1 clone, Dako North America, Inc.

Data from comparisons between 39 XY*O males and 40 XY MF1 males, and pharmacologic manipulation of steroid sulfatase activities consistently support the role of steroid sulfatase in attention as assessed by 5CSRTT [64]. Interestingly, however, 39 XY*O males exhibit reduced premature responses in the 5CSRTT, suggesting a lower level of impulsivity compared to 40 XY MF1 males [64]. Moreover, this website using a recently developed paradigm of the stop-signal reaction time task for evaluating behavioral inhibition and impulsivity [65•], Davies et al. demonstrated that genetic and pharmacologic inhibition of steroid sulfatase resulted in enhanced response control

[66••]. These studies provide evidence that the genetic basis of inattention and impulsivity is dissociable, and support the use of 39 XY*O mice as a genetic model of ADHD without impulsivity. Studies with BDX recombinant inbred strains provide strong evidence for http://www.selleckchem.com/products/FK-506-(Tacrolimus).html the importance of gene–gene

interactions in attention and impulsivity 67•• and 68••]. Behavioral phenotypes in impulsivity and attention analyzed by the 5CSRTT and PPI tests surpass those of the C57BL/6J and DBA/2J founders. A forward genetic approach utilizing BDX recombinant inbred strains led to the identification of the developmental roles of neuregulin-3 (Nrg3) in the mouse medial prefrontal cortex in regulating impulsive activity [68••]. Nrg3-KO mice have decreased impulsivity. Viral overexpression of Nrg3 in the medial prefrontal cortex of Clostridium perfringens alpha toxin wild-type mice increases impulsivity, but does not rescue Nrg3-KO mouse phenotypes [68••]. Thus, the Nrg3 expression level is likely crucial. Nrg-3 binds to the extracellular domain of the ErbB4 receptor tyrosine kinase [69], and is likely associated with attention deficits in humans [70]. ADHD mouse genetic models have become substantially diversified, reflecting the progress in human genetics and supporting the

notion that ADHD has a polygenic nature. Further efforts are needed to establish novel genetic models. For example, some representative genes, such as T-cadherin and metabotropic glutamate receptor 5, which are strongly supported by human genetic studies, have not been experimentally evaluated. Data from BDX recombinant inbred strains clearly indicate the importance of gene-gene interactions. Neuronal mechanisms for attention and impulse control domains are complex and are supported by large neuronal networks. Behavioral phenotypes of current mouse models have been analyzed to different extents, and available tests for assessing attention and impulsivity remain suboptimal. Future studies of mouse models using refined behavioral tests and careful examination of circuit activities will enhance our understanding of the circuit mechanisms underlying attention and impulsivity.

To avoid

these and related issues, the integrated judgement-based approach used here has also been adopted in other jurisdictions, such as a pilot for the World Ocean Assessment (Feary et al., 2014), to provide a defendable framework for the rapid assessment of data-poor ocean ecosystems. This paper reports the combined personal judgements of the scientists who contributed to the assessments—a diverse and highly experienced group of independent experts with relevant backgrounds from various tertiary and science institutions. Their judgements were developed in a structured and peer-group contestable process and the findings are based on all available Forskolin supplier data and knowledge. Although substantial uncertainty remains around the accuracy of the findings for many of the environmental components, the breadth and robustness of this consultative process provides a basis for development of improved national-scale marine policies and strategies that focus on the intrinsic attributes

of ecosystem structure and function as well as gaps in knowledge. Without such a comprehensive approach, national-scale assessments risk becoming simply reports that re-confirm the technical detail of what is already known GSK2118436 chemical structure rather than a systematic and balanced analysis of the performance of ocean environments as a whole. The national marine condition assessment process and the SoE 2011 report was funded and supported by DSEWPaC (now the Department of the Environment), and the support and leadership provided by the divisions Thalidomide of DSEWPaC, and members of the independent committee established to prepare State of the Environment Australia 2011 (http://www.environment.gov.au/soeSoEC, 2011) is gratefully acknowledged. Earlier drafts of this paper have been improved by review, comment and inputs from Nancy Dahl Tacconi, Boon Lim,

Ilse Keissling, Nicole Coombe and Carolyn Armstrong (all the Department of the Environment) and external reviewers of the draft manuscript: Kirstin Dobbs, Great Barrier Reef Marine Park Authority; Richard Kenchington, University of Wollongong, and Ian Perry, Fisheries and Oceans Canada. The willingness and commitment of the 40 experts who openly contributed to the workshops and the grading process is also very gratefully acknowledged—these experts are individually identified in Ward, 2011. The workshops were facilitated by Richard Stoklosa (E-Systems Pty Ltd, Tasmania). The interpretations of the SoE data and the opinions in this paper remain those of the author alone, and do not necessarily represent official Australian Government policy, or the specific view of any single expert who contributed information.

As a result, improvements in the accuracy of attenuation correction in the abdomen are considered a work in progress. Another challenge in attenuation correction in PET–MRI is to account

for attenuation due to the radiofrequency Apitolisib order (RF) coil (required for all MR acquisitions) which has been shown to adversely affect the quantitative accuracy of PET emission data by significant amounts [46]. As the coil does not appear in the MR image, its attenuation must be accounted for separately in an MR-based approach. One recent study provided evidence that a using a high-exposure CT to obtain a model of (in this particular case) a head coil could be used in a model-based correction that gave attenuation-corrected PET images that

were comparable to the reference PET–CT reconstruction [47]. The authors noted that if there were errors in the positioning of the coil (on the order of a few millimeters), then artifacts emerged in the reconstructed PET image. Tellman et al. found similar results on the importance of coil alignment [48]. find more Though challenging, careful engineering should adequately address this problem, as the geometry and composition of MR RF coils can be fixed for most, though not all, coil designs. Integration of PET/MR systems with advanced flexible coil designs, or endoscopic coils such as endorectal coils for prostate imaging, may require additional materials engineering work in reducing Avelestat (AZD9668) net attenuation of such designs, or real-time feedback on their location. Besides the use of MR data to correct for the effect of attenuation on PET data, simultaneously acquired MR images also offer the potential to improve PET images by providing anatomical information that can be incorporated into the PET image reconstruction process. Statistical reconstruction

algorithms are replacing filtered backprojection as the method of choice for generating PET images from coincidence data, primarily because they provide a framework in which the physical properties of the data collection process can be modeled [49]. We expect different tissue types to exhibit different tracer uptake levels, so knowledge of tissue boundaries can be incorporated into the PET image reconstruction process to reduce blurring at those boundaries [50], [51] and [52]. While these methods have been applied to PET–CT as well as retrospectively co-registered PET–MRI data, simultaneously acquired MRI data offer superior soft-tissue contrast with the most accurate spatial registration. There are three major types of motion that must be considered during PET acquisition: gross motion (e.g., head movement or subtle patient repositioning due to discomfort), periodic movement (e.g., cardiac and respiratory motion), and internal shifting and distortion in the pelvic and abdominal regions (e.g., peristalsis).

(1), (2), (3), (4), (5) and (6)) applied on appropriate SHI sequences. The same theorem with the Gamma pdf of flows can be applied to estimate the above parameters on monthly time scale. In both situations, μ, cv, and ρ1 can be used to provide reliable estimates of E(LT) and E(MT) at the truncation level equivalent to the median

flow level over a period of T-year. The drought analysis on weekly time scale becomes complex because of the involved underlying dependence structure and thus the second order Markov chain models are considered for which there is a paucity of close form equations for estimating the second order conditional Stem Cell Compound Library probabilities, viz. qqq and qqp. Therefore, the historical flow records are used to estimate these parameters by the counting method involving BIBW2992 both the non-standardized flow series and appropriate SHI sequences. Potentially,

there are 3 values (based on the annual, monthly, and weekly time scales) of E(LT) for a T-year drought and consequently 3 values of the expected deficit-volumes, E(DT) that need to be considered for the assessment of volumetric-storage [E(DT) = σE(MT)]. A logical question that naturally arises as to which one of them should be used for planning the drought mitigation measures. To elucidate the point, the case of Torrent river, Canada (station NF02YC001) with the following statistical properties is considered: mean flow equal to 24.50 m3/s; σ equal to 3.68 m3/s (annual), 12.50 m3/s (monthly averaged value), 17.15 m3/s (weekly averaged value); ρ1 equal to 0.0 (annual, assumed as 0.0 in view of negligible dependence), 0.19 (monthly), and 0.73 (weekly). On annual, monthly, and weekly time scales, the values of cv ( Table 1 and Table 2) are respectively 0.15, 0.51 and 1.12 for the computations of E(LT). The values of qq, qqq and qqp were estimated as 0.76 and 0.84 and 0.24 at the median level (i.e. q = 0.5 and SHI0 = −0.32). Using the above statistics, it can be estimated that a 50-year drought is likely to continue for 5 years or 10 months or 33 weeks respectively

when analyzed based on annual, monthly, Niclosamide and weekly time scales (by plugging the values of parameters in Equations (1), (2), (3), (4), (5), (6), (7) and (8)). The corresponding values of drought magnitudes can be computed as 0.58 (=3.68 × 5 × c1) billion m3, or 0.32 (=12.50 × 10 × c2) billion m3 or 0.24 (=17.15 × 0.69 × 33 × c3) billion m3. Note c1 (=31.5 × 106), c2 (=2.95 × 106) and c3 (=0.605 × 106) are conversion constants to covert the annual, monthly and weekly flow rates into volumes. It may be borne in mind that for annual and monthly droughts drought intensity, E(I) equal to 1 and for weekly drought E(I) equal to 0.69 (Eq. (6), z0 = SHI0 = −0.32 and corresponding q for normal pdf is 0.37) for use in the relationship E(MT) = E(I) × E(LT).

45). At m  =0 effort is equal to the pure open access case, E=  1−c  . It can be selleck seen that the value of c   determines the maximum of E  , in fact this is the maximum sustainable yield effort Empamsy=1/4c, whereas the value of γ influences the location of the maximum. The reason for this is that the speed

at which the fish migrate influences the stock size within the MPA, and for slow moving fish there will be a sufficiently large stock within the MPA to provide high spillover levels at low MPA sizes. As the MPA increases in size, the low speed of migration ensures that more of the fish is retained within the reserve, thereby reducing the spillover effect. For species with a high γ, the stock build-up within the reserve is too low to provide high levels of spillover when the MPA is small. As the MPA increases, so does the stock level and the high speed of migration

ensures that large spillover effects are generated even at large MPA sizes. With respect to employment, it has been demonstrated that for the cases when the MPA can realize MSY, both fishing and processing employment increase with MPA size up to the MSY reserve. However, further increase will reduce both effort this website and processing employment. With constant price of harvest and cost of effort no resource rent, consumer surplus (CS) and producer surplus (PS), are generated in the analyses above. Then, from an economic point of view, why bother with establishing MPAs if no economic rent is generated? There are at least two answers to this. First, actual fishing fleets often display heterogeneous vessels and costs – implying intra-marginal rent in open-access fisheries [33] – to be discussed below. Second, actual fish markets often display downward sloping demand and the possibilities of CS. This will now be discussed. The increased harvest following the creation of an MPA (see above) combined with a downward sloping demand curve allows for the creation of CS. Pezzey et al. [22] mention additionally,

in the case of marine reserves, the ID-8 possibility of a shift in demand caused by “more desirable fish” and in supply, caused by “more easily catchable fish”. Now, investigate the case of consumer surplus to see how this changes the previous conclusions about zero economic rent. Fig. 5 shows the backward bending long-run open-access supply curve as a function of the fish price, assuming all other parameters being constant, [34] and [35].6 With a downward sloping demand curve for harvest assume that there is a unique stable equilibrium at overall open-access, with price of harvest po and harvest Y at O 7 ( Fig. 5). With an MPA the backward bending supply curve shifts to the right and upwards. The MSY supply (equal to 0.25) is the same for all three curves. Demand and supply conditions in Fig. 5 have been chosen such that HZ open-access is close to realizing MSY for MPA size m=0.75. In this case, the CS equals the triangle pmMp, which is significantly greater than the pre-MPA CS triangle poOp.

The nerve was drawn into a suction electrode for stimulation with 100 μs supramaximal stimuli at 0.5 Hz using a Grass SD9 stimulator. The membrane potential was also recorded and used to

correct amplitudes and areas of MEPPs and EPPs to a standard resting potential of −35 mV and for non-linear summation when EPP amplitude exceeded 10% of the driving force assuming a reversal potential of 0 mV for synaptic current (McLachlan and Martin, 1981). Quantal content was calculated by the direct method as (EPP/MEPP), where the average MEPP amplitude was the mean of at least 40 events. After a control period, the bath perfusion was stopped, PhKv toxin was added directly to the bath at a final concentration of 200 nM and the effects were measured 10 min Palbociclib molecular weight later. Control recordings without toxin were time matched to detect any time-dependent run-down of the preparation. Adult rat heart cells were prepared by standard methods, as previously

described (Guatimosim et al., 2001). Briefly, rats of either sex weighing between 200 and 300 g were killed by decapitation. The hearts were rapidly removed and perfused via the Langendorff method with Ca2+-free modified Tyrode solution until the blood was washed out. Hearts were then perfused with Tyrode solution containing 50 mM CaCl2 along with 1.4 mg/ml collagenase (type buy MDV3100 2; Worthington, Lakewood, NJ) and 0.04 mg/ml protease (type XIV; Sigma, St. Louis, MO) until they were soft. The hearts were removed from the perfusion apparatus,

minced into; 1 mm chunks, and stirred for 4 min in Tyrode solution containing 50 mM CaCl2, 0.7 mg/ml collagenase, and 0.02 mg/ml protease. Cells were filtered through a 200 mm mesh to remove tissue chunks, and extracellular Ca2+ concentration was raised to 0.5 mM C-X-C chemokine receptor type 7 (CXCR-7) over 10 min through three centrifuge cycles. Cells were stored in DMEM until they were used (within 4 h). Following incubation with 6.6 μM fluor-4 AM (Molecular Probes) for 30 min, isolated cardiomyocytes were field-stimulated (1 Hz) in control solution or solution containing PhKv (250 nM) at 1 Hz. Data were acquired under steady-state conditions with Zeiss LSM 510 META confocal microscope (CEMEL, ICB-UFMG). All experiments were performed at room temperature. The Ca2+ level was reported as F/F0, where F0 is the resting Ca2+ fluorescence inside the cell and F is the peak fluorescence signal. Action potentials were measured as described previously (Lara et al., 2010). The pipette solution contained (in mM): 110 K+-aspartate, 20 KCl, 8 NaCl, 1 MgCl2 , 1 CaCl2,10 HEPES, 10 EGTA (pH = 7.2 with KOH). The superfusion solution contained 140 NaCl, 1 MgCl2, 0.33 NaH2PO4, 10 HEPES, 10 glucose, 1.8 CaCl2 and 5 KCl; pH 7.4. Pclamp 8.0, Origin (version 8.0) and IDL (Research Systems) were used for data analysis. To obtain electrocardiographic tracings, the rats (n = 4) were anesthetized with 2.