ATP-sensitive potassium channel subcellular trafficking during ischemia, reperfusion, and preconditioning

Ischemic preconditioning is an endogenous cardioprotective mechanism in which short periods of ischemia and reperfusion provide protection when given before a subsequent ischemic event. Early mechanistic studies showed ATP-sensitive potassium (KATP) channels to play an important role in ischemic preconditioning. KATP channels link intracellular energy metabolism to membrane excitability and contractility. It is thought that KATP channels provide a cardioprotective role during ischemia by inducing action potential shortening, reducing an excessive Ca^2+ influx, and by preventing arrhythmias. However, the mechanisms by which KATP channels protect during ischemic preconditioning are not known. In this study, we investigated a novel potential mechanism in which alterations in subcellular KATP channel trafficking during ischemia and ischemic preconditioning may result in altered levels of surface channel density, and therefore, a greater degree of cardioprotection. In the optimization of our experiments, we compared various antibodies for their specificity and sensitivity for channel subunit detection in immunoblotting. In addition, we examined the effects of varying salt concentrations during tissue homogenization in order to determine the optimal conditions for protein isolation. Furthermore, we examined the effect of heating the samples prior to SDS-PAGE for improved detection of channel proteins by immunoblotting. The subcellular trafficking of some membrane proteins is altered by ischemia. For example, the glucose transporter, Glut4, translocates from endosomal compartments to the sarcolemma (Sun, Nguyen, DeGrado, Schwaiger, & Brosius, 1994). Conflicting data exists regarding the effects of ischemia on KATP channel subcellular trafficking and the regulation of KATP channel surface density (Edwards et al., 2009 and Bao, Hadjiolova, Coetzee, & Rindler, 2011). We therefore, sought to test our hypothesis that KATP channels are internalized from the surface of cardiomyocytes to endosomal compartments during ischemia, and this internalization can be reduced and/or reversed by ischemic preconditioning. We subjected isolated Langendorff-perfused mouse hearts to ischemia, reperfusion, or ischemic preconditioning events and measured the density of KATP channels in the sarcolemmal and endosomal compartments. We also determined the degree of injury by staining heart slices with triphenyltetrazolium chloride and compared infarct sizes between hearts subjected to ischemia and ischemic preconditioning. Our data demonstrated that KATP channels are, in fact, internalized during ischemia and that reperfusion led to a slow recovery of surface KATP channel density. Interestingly, ischemic preconditioning reduced the size of infarcts induced by ischemia and also prevented the ischemia-induced decrease of KATP channel surface density, thereby, contributing to cardioprotection

Cognitive radio network in vehicular ad hoc network (VANET): a survey

Cognitive radio network and vehicular ad hoc network (VANET) are recent emerging concepts in wireless networking. Cognitive radio network obtains knowledge of its operational geographical environment to manage sharing of spectrum between primary and secondary users, while VANET shares emergency safety messages among vehicles to ensure safety of users on the road. Cognitive radio network is employed in VANET to ensure the efficient use of spectrum, as well as to support VANET’s deployment. Random increase and decrease of spectrum users, unpredictable nature of VANET, high mobility, varying interference, security, packet scheduling, and priority assignment are the challenges encountered in a typical cognitive VANET environment. This paper provides survey and critical analysis on different challenges of cognitive radio VANET, with discussion on the open issues, challenges, and performance metrics for different cognitive radio VANET applications

Performance modelling of adaptive VANET with enhanced priority scheme

In this paper, we present an analytical and simulated study on the performance of adaptive vehicular ad hoc networks (VANET) priority based on Transmission Distance Reliability Range (TDRR) and data type. VANET topology changes rapidly due to its inherent nature of high mobility nodes and unpredictable environments. Therefore, nodes in VANET must be able to adapt to the ever changing environment and optimize parameters to enhance performance. However, there is a lack of adaptability in the current VANET scheme. Existing VANET IEEE802.11p’s Enhanced Distributed Channel Access; EDCA assigns priority solely based on data type. In this paper, we propose a new priority scheme which utilizes Markov model to perform TDRR prediction and assign priorities based on the proposed Markov TDRR Prediction with Enhanced Priority VANET Scheme (MarPVS). Subsequently, we performed an analytical study on MarPVS performance modeling. In particular, considering five different priority levels defined in MarPVS, we derived the probability of successful transmission, the number of low priority messages in back off process and concurrent low priority transmission. Finally, the results are used to derive the average transmission delay for data types defined in MarPVS. Numerical results are provided along with simulation results which confirm the accuracy of the proposed analysis. Simulation results demonstrate that the proposed MarPVS results in lower transmission latency and higher packet success rate in comparison with the default IEEE802.11p scheme and greedy scheduler scheme

Validation of a Novel Traditional Chinese Medicine Pulse Diagnostic Model Using an Artificial Neural Network

In view of lacking a quantifiable traditional Chinese medicine (TCM) pulse diagnostic model, a novel TCM pulse diagnostic model was introduced to quantify the pulse diagnosis. Content validation was performed with a panel of TCM doctors. Criterion validation was tested with essential hypertension. The gold standard was brachial blood pressure measured by a sphygmomanometer. Two hundred and sixty subjects were recruited (139 in the normotensive group and 121 in the hypertensive group). A TCM doctor palpated pulses at left and right cun, guan, and chi points, and quantified pulse qualities according to eight elements (depth, rate, regularity, width, length, smoothness, stiffness, and strength) on a visual analog scale. An artificial neural network was used to develop a pulse diagnostic model differentiating essential hypertension from normotension. Accuracy, specificity, and sensitivity were compared among various diagnostic models. About 80% accuracy was attained among all models. Their specificity and sensitivity varied, ranging from 70% to nearly 90%. It suggested that the novel TCM pulse diagnostic model was valid in terms of its content and diagnostic ability

Proteomic analysis identifies interleukin 11 regulated plasma membrane proteins in human endometrial epithelial cells in vitro

<p>Abstract</p> <p>Background</p> <p>During the peri-implantation period, the embryo adheres to an adequately prepared or receptive endometrial surface epithelium. Abnormal embryo adhesion to the endometrium results in embryo implantation failure and infertility. Endometrial epithelial cell plasma membrane proteins critical in regulating adhesion may potentially be infertility biomarkers or targets for treating infertility. Interleukin (IL) 11 regulates human endometrial epithelial cells (hEEC) adhesion. Its production is abnormal in women with infertility. The objective of the study was to identify IL11 regulated plasma membrane proteins in hEEC <it>in vitro </it>using a proteomic approach.</p> <p>Methods</p> <p>Using a 2D-differential in-gel electrophoresis (DIGE) electrophoresis combined with LCMS/MS mass spectrometry approach, we identified 20 unique plasma membrane proteins differentially regulated by IL11 in ECC-1 cells, a hEEC derived cell line. Two IL11 regulated proteins with known roles in cell adhesion, annexin A2 (ANXA2) and flotillin-1 (FLOT1), were validated by Western blot and immunocytochemistry in hEEC lines (ECC-1 and an additional cell line, Ishikawa) and primary hEEC. Flotilin-1 was further validated by immunohistochemistry in human endometrium throughout the menstrual cycle (<it>n = 6-8/cycle</it>).</p> <p>Results</p> <p>2D-DIGE analysis identified 4 spots that were significantly different between control and IL11 treated group. Of these 4 spots, there were 20 proteins that were identified with LCMS/MS. Two proteins; ANXA2 and FLOT1 were chosen for further analyses and have found to be significantly up-regulated following IL11 treatment. Western blot analysis showed a 2-fold and a 2.5-fold increase of ANXA2 in hEEC membrane fraction of ECC-1 and Ishikawa cells respectively. Similarly, a 1.8-fold and a 2.3/2.4-fold increase was also observed for FLOT1 in hEEC membrane fraction of ECC-1 and Ishikawa cells respectively. <it>In vitro</it>, IL11 induced stronger ANXA2 expression on cell surface of primary hEEC and ECC-1 whilst, the lipid-raft protein FLOT1 demonstrated punctate staining in the apical surface of ECC-1 plasma membranes and was upregulated in the epithelium in the receptive phase of the menstrual cycle (p lower or equal 0.05).</p> <p>Conclusions</p> <p>This is the first study to use a proteomics approach to identify hEEC plasma membrane proteins that may be useful as infertility markers or pharmacological targets for fertility regulation.</p

Roles and recommendations from primary care physicians towards managing low-risk breast cancer survivors in a shared-care model with specialists in Singapore-a qualitative study.

BackgroundBreast cancer is prevalent and has high cure rates. The resultant increase in numbers of breast cancer survivors (BCS) may overwhelm the current oncology workforce in years to come. We postulate that primary care physicians (PCPs) could play an expanded role in comanaging survivors, provided they are given the appropriate tools and training to do so.ObjectiveTo explore the perspectives of PCPs towards managing BCS in a community-based shared-care programme with oncologists.MethodsEleven focus groups and six in-depth interviews were conducted with seventy PCPs recruited by purposive sampling. All sessions were audio-recorded, transcribed verbatim and coded by three independent investigators. Thematic data analysis was performed and the coding process facilitated by NVivo 12.ResultsMajority of PCPs reported currently limited roles in managing acute and non-cancer issues, optimizing comorbidities and preventive care. PCPs aspired to expand their role to include cancer surveillance, risk assessment and addressing unmet psychosocial needs. PCPs preferred to harmonize cancer survivorship management of their primary care patients who are also BCS, with defined role distinct from oncologists. Training to understand the care protocol, enhancement of communication skills, confidence and trust were deemed necessary. PCPs proposed selection criteria of BCS and adequacy of their medical information; increased consultation time; contact details and timely access to oncologists (if needed) in the shared-care programme.ConclusionsPCPs were willing to share the care of BCS with oncologists but recommended role definition, training, clinical protocol, resources and access to oncologist's consultation to optimize the programme implementation

The protection of Victoria Harbour in Hong Kong : an analysis of civic engagement strategies

published_or_final_versionPolitics and Public AdministrationMasterMaster of Public Administratio

Comparison of crude extract from durio zibethinus M. (durian) leaf waste via ultrasound-assisted extraction and accelerated solvent extraction: antioxidant activity and cytotoxicity

The objective of this study was to compare the antioxidant activity and cytotoxicity of Durio zibethinus M. (Durian) leaf extract from two extraction methods. Ultrasound-assisted extraction and Accelerated-solvent extraction were used to produce crude extract. The results revealed that UAE achieved 3× higher in total phenolic content in the leaf extract compared to ASE. DPPH radical scavenging activity was 4.6× higher in leaf extract from ASE. No significant differences reported in ferric reducing power, and total flavonoid content of the leaf extract between the two methods. Cytotoxicity via MTT assay demonstrated no significant differences in cell viability upon exposure to the leaf extract from both methods. This suggested that they were appropriate in producing Durio zibethinus M. leaf extract for end use application in food related product. Both ensured similar level of safety in Durio zibethinus M. leaf extract as a new potential ingredient for the food industry