Selectively inhibition of oxalate-stimulated Ca2+ transport by cyclopiazonic acid and thapsigargin in smooth muscle microsomes

45Ca2+ uptake and efflux studies were performed on membranes prepared from dog mesenteric artery and rat vas deferens. Oxalate-stimulated, ATP-dependent Ca2+ uptake in microsomal vesicles, a property characteristic of sarcoplasmic reticulum, was completely inhibited in a concentration-dependent manner by cyclopiazonic acid (0.1-30 (iM) and thapsigargin (10 nM - 10 |xM). Using discontinuous sucrose gradient centrifugation, rat vas deferens microsomes were separated into two fractions, one enriched in plasma membrane (F2), the other enriched in sarcoplasmic reticulum (F3). The F3 fraction had a major increase in Ca2+ uptake in the presence of oxalate, which was completely inhibited by either cyclopiazonic acid or thapsigargin. In the F2 fraction Ca2+ uptake in the presence of oxalate was lower than in F3 and was not completely inhibited by thapsigargin and cyclopiazonic acid. Instead, the F2 fraction had a thapsigargin-insensitive and cyclopiazonic acid insensitive, saponin-sensitive component of uptake, which probably represents Ca2+ uptake by plasma membrane. In the absence of oxalate, the inhibition of Ca2+ uptake by saponin and cyclopiazonic acid or thapsigargin was additive in the F2 and F3 fractions, suggesting that cyclopiazonic acid and thapsigargin selectively inhibited sarcoplasmic reticulum derived Ca2+ uptake and did not affect plasma membrane derived Ca2+ uptake. Measurement of the initial rate of Ca2+ uptake in the presence and absence of oxalate by rat vas deferens microsomes demonstrated selective inhibition of oxalate-stimulated Ca2+ uptake by cyclopiazonic acid and thapsigargin. Ca2+ efflux from rat vas deferens microsomes actively loaded with 45Ca2+ either in the presence or the absence of oxalate was not increased by cyclopiazonic acid or thapsigargin, showing that the inhibition of Ca2+ accumulation was not due to an increase in Ca2+ efflux. In both rat vas deferens and dog mesenteric artery, the maximal inhibitory effects of cyclopiazonic acid developed rapidly, whereas for maximal inhibition thapsigargin required pretreatment of microsomes prior to measurement of Ca2+ uptake. In rat vas deferens microsomes the inhibitory effects of cyclopiazonic acid could be quickly and completely reversed, whereas the effects of thapsigargin were not easily reversed. Collectively, these results suggest selectivity of cyclopiazonic acid and thapsigargin for the sarcoplasmic reticulum Ca2+ pump. Their selective inhibitory properties and differences in onset and offset of inhibition make cyclopiazonic acid and thapsigargin useful pharmacological tools in the study of the physiological and pathophysiological roles of the sarcoplasmic reticulum Ca2+ pump in regulating smooth muscle Ca2+.published_or_final_versio

Regulation of vascular tone: cross-talk between sarcoplasmic reticulum and plasmalemma

Selected topics on the roles of sarcoplasmic reticulum (SR) in the control of vascular smooth muscle (VSM) tone are briefly reviewed with particular reference to the regulation of cytosolic concentration of free calcium ions, [Ca2+]i. Although morphological evidence and subcellular membrane studies indicate a relatively meager quantity of SR in VSM and of endoplasmic reticulum (ER) in endothelial cells (ECs) compared with skeletal muscle and cardiac muscle, contractility studies suggest that vascular tone is, to a large extent, regulated by the intracellular Ca2+ stores in smooth muscle and endothelial cells. Cytosolic Ca2+ levels control myosin light chain phosphorylation and contraction in VSM and activation of NO synthase and phospholipase A2 in ECs to regulate nitric oxide (NO) and prostaglandin I2 formation. Understanding of the importance of SR or ER in modulating the [Ca2+]i in VSM and ECs has been further advanced as a result of the new development and refinement of biophysical techniques in the measurement of cellular Ca2+ concentrations and ion currents, such as fluorescent Ca2+ indicators and patch-clamp techniques. Experimental evidence has accumulated in support of the existence of cross-talk between SR-ER and the plasma membrane (PM). Novel pharmacological tool drugs selective for the SR-ER Ca2+ pump, such as thapsigargin and cyclopiazonic acid, as well as for SR-ER Ca2+ channels, such as ryanodine (for the Ca(2+)-induced Ca2+ release channel) and inositol polyphosphates and heparin (for the inositol-1,4,5-trisphosphate activated Ca2+ channel), together with the use of blockers for selective PM Ca2+ channels have enabled better formulation and elucidation of the mechanisms of cross-talk between SR-ER and PM.(ABSTRACT TRUNCATED AT 250 WORDS)published_or_final_versio

Concordance between side-stream end-tidal carbon dioxide and arterial carbon dioxide partial pressure in respiratory service setting

OBJECTIVE: To explore the correlation and concordance between end-tidal carbon dioxide and arterial carbon dioxide partial pressure, and confirm the experience of the general consensus among service environments. DESIGN: A prospective cross-sectional analysis. SETTING: Two respiratory service units in Hong Kong. PARTICIPANTS: Two hundred respiratory patients were recruited, in whom 219 sets of observations were recorded. Patients deemed to require arterial blood gas determination also had their end-tidal carbon dioxide partial pressure measured at that time, using two LifeSense LS1-9R Capnometers. MAIN OUTCOME MEASURES: The agreement of end-tidal carbon dioxide partial pressure and arterial carbon dioxide partial pressure was studied by correlation coefficients, mean and standard deviation of their difference, and the Bland-Altman plot. RESULTS: Overall, the correlation was low and insignificant (r=0.1185, P=0.0801). The mean of the difference was 7.2 torr (95% confidence interval, 5.5-8.9) and significant (P<0.001). The limits of agreement by Bland-Altman analysis were -18.1 to 32.5 torr, which were too large to be acceptable. In the sub-group on room air, the mean difference was reduced to 2.26 torr, the correlation between end-tidal carbon dioxide partial pressure and arterial carbon dioxide partial pressure was 0.2194 (P=0.0068), though statistically significant, the extent of correlation was still low. CONCLUSION: End-tidal carbon dioxide partial pressure did not show significant correlation or concordance with arterial carbon dioxide partial pressure, especially when supplemental oxygen was used. End-tidal carbon dioxide partial pressure currently cannot replace arterial blood gas measurement as a tool for monitoring arterial carbon dioxide partial pressure. Possible reasons for the discrepancy with previous studies include small sample size in previous studies, lack of research facilities in service settings, and publication bias against negative studies.published_or_final_versio

XMM-Newton and NuSTAR Observations of the Compact Millisecond Pulsar Binary PSR J1653–0158

We have presented the first joint XMM-Newton and NuSTAR analysis of the millisecond pulsar (MSP) binary PSR J1653−0158. The 75 minute orbital period inferred from optical and gamma-ray observations together with the 1.97 ms pulsation in the gamma-rays indicate that this system is the most compact Black Widow MSP system known to date. The orbital period was not detected in the XMM-Newton and NuSTAR data, probably due to insufficient photon counts obtained in the observations. Fitting the joint X-ray spectrum of PSR J1653−0158 with a power law gives a photon index Γ = 1.71 ± 0.09. The X-ray luminosity of the source in the (0.2–40) keV band is deduced to be 1.18 × 1031 erg s−1, for an adopted distance of 0.84 kpc. We have shown that the broadband X-ray spectrum can be explained by synchrotron radiation from electrons accelerated in the intrabinary shock, and the gamma-rays detected in the Fermi data are curvature radiations from electrons and positrons in the pulsar magnetosphere. Our kinematic analysis of the Tidarren systems PSR J1653–0158 and PSR J1311–3430 indicates that the two Tidarren systems are likely to have originated in the Galactic disk

Consumption of dried fruit of Crataegus pinnatifida (hawthorn) suppresses high-cholesterol diet-induced hypercholesterolemia in rats

Author name used in this publication: Mabel Yin-Chun YauAuthor name used in this publication: Peter Hoi-Fu Yu2009-2010 > Academic research: refereed > Publication in refereed journalAccepted ManuscriptPublishe

Evaluation of anti-oxidant capacity of root of Scutellaria baicalensis Georgi, in comparison with roots of polygonum multiflorum thunb and Panax ginseng CA Meyer

Author name used in this publication: Jian-Hong WuAuthor name used in this publication: Alice Lai-Shan AuAuthor name used in this publication: Peter Hoi-Fu Yu2009-2010 > Academic research: refereed > Publication in refereed journalAccepted ManuscriptPublishe

Quantitative Proteomic Analysis of Simian Primary Hepatocytes Reveals Candidate Molecular Markers for Permissiveness to Relapsing Malaria Plasmodium cynomolgi.

A major obstacle impeding malaria research is the lack of an in vitro system capable of supporting infection through the entire liver stage cycle of the parasite, including that of the dormant forms known as hypnozoites. Primary hepatocytes lose their liver specific functions in long-term in vitro culture. The malaria parasite Plasmodium initiates infection in hepatocyte. This corresponds to the first step of clinically silent infection and development of malaria parasite Plasmodium in the liver. Thus, the liver stage is an ideal target for development of novel antimalarial interventions and vaccines. However, drug discovery against Plasmodium liver stage is severely hampered by the poor understanding of host-parasite interactions during the liver stage infection and development. In this study, tandem mass tag labeling based quantitative proteomic analysis is performed in simian primary hepatocytes cultured in three different systems of susceptibility to Plasmodium infection. The results display potential candidate molecular markers, including asialoglycoprotein receptor, apolipoproteins, squalene synthase, and scavenger receptor B1 (SR-BI) that facilitate productive infection and full development in relapsing Plasmodium species. The identification of these candidate proteins required for constructive infection and development of hepatic malaria liver stages paves the way to explore them as therapeutic targets

Tetramethylpyrazine attenuates spinal cord ischemic injury due to aortic cross-clamping in rabbits

BACKGROUND: Lower limb paralysis occurs in 11% of patients after surgical procedure of thoracic or thoracoabdominal aneurysms and is an unpredictable and distressful complication. The aim of this study was to investigate the effects of tetramethylpyrazine (TMP), an intravenous drug made from traditional Chinese herbs, on the neurologic outcome and hisotpathology after transient spinal cord ischemia in rabbits. METHODS: Forty-five male New Zealand white rabbits were anesthetized with isoflurane and spinal cord ischemia was induced for 20 min by infrarenal aortic occlusion. Animals were randomly allocated to one of five groups (n = 8 each). Group C received no pharmacologic intervention. Group P received intravenous infusion of 30 mg·kg(-1) TMP within 30 min before aortic occlusion. Group T(1), Group T(2) and Group T(3) received intravenous infusion of 15, 30 and 60 mg·kg(-1) TMP respectively within 30 min after reperfusion. In the sham group (n = 5), the animals underwent the same procedures as the control group except infrarental aortic unocclusion. Neurologic status was scored by using the Tarlov criteria (in which 4 is normal and 0 is paraplegia) at 4 h, 8 h, 12 h, 24 h, and 48 h after reperfusion. All animals were sacrificed at 48 h after reperfusion and the spinal cords (L(5)) were removed immediately for histopathologic study. RESULTS: All animals in the control group became paraplegic. Neurologic status and histopathology (48 h) in the Groups P, T(2) and T(3) were significantly better than those in the control group (P < 0.05). There was a strong correlation between the final neurologic scores and the number of normal neurons in the anterior spinal cord (r = 0.776, P < 0.01). CONCLUSION: Tetramethylpyrazine significantly reduces neurologic injury related to spinal cord ischemia and reperfusion after aortic occlusion within a certain range of dose

Validation of a prognostic scoring system for locally recurrent nasopharyngeal carcinoma treated by stereotactic radiosurgery

<p>Abstract</p> <p>Background</p> <p>Selection of patients with local failure of nasopharyngeal carcinoma (NPC) for appropriate type of salvage treatment can be difficult due to the lack of data on comparative efficacy of different salvage treatments. The purpose of the present study was to validate a previously published prognostic scoring system for local failures of NPC treated by radiosurgery based on reported results in the literature.</p> <p>Methods</p> <p>A literature search yielded 3 published reports on the use of radiosurgery as salvage treatment of NPC that contained sufficient clinical information for validation of the scoring system. Prognostic scores of 18 patients from these reports were calculated and actuarial survival rates were estimated and compared to the original cohort used to design the prognostic scoring system. The area under the receiver operating characteristic curve was also determined and compared between the current and original patient groups.</p> <p>Results</p> <p>The calculated prognostic scores ranged from 0.32 to 1.21, with 15 patients assigned to the poor prognostic group and 3 to the intermediate prognostic group. The actuarial 3-year survival rates in the intermediate and poor prognostic groups were 67% and 0%, respectively. These results were comparable to the observed 3-year survival rates of 74% and 23% in the intermediate and poor prognostic group in the original reports. The area under the receiver operating characteristic curve for the current patient group was 0.846 which was similar to 0.841 in the original group.</p> <p>Conclusion</p> <p>The previously published prognostic scoring system demonstrated good prediction of treatment outcome after radiosurgery in a small group of NPC patients with poor prognosis. Prospective study to validate the scoring system is currently being carried out in our institution.</p

An apparatus for studying spallation neutrons in the Aberdeen Tunnel laboratory

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