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Groundwater samples (n=15) were collected from various sites of Shah Faisal Town from a variable depth range (70-400 feet). The TDS varied from 766.08 to 5490.24 mg/L with a mean of 3128.16 mg/L. Circum-neutral pH (6.84-7.56) is evident within the WHO guidelines (6.5-8.5) for drinking. All the samples have objectionable (> 500 mg/L) level of hardness content where it shows a wide range (600-1800 mg/l; mean: 1200 mg/l). The samples ' sodium content (Range: 200-1223; mean: 711.5 mg/L) exceeded WHO limit (200 mg/L). Calcium and magnesium concentrations varied between 40-240 and 92.34-340.2 mg/l with a mean of 220 mg/l and 216.27 mg/l, respectively. Both these elements exceed about 5 times the corresponding permissible limits set by WHO for drinking water. On the other hand, two third samples show elevated bicarbonate content (Range: 210-500; Mean: 355 mg/l). Similarly, nitrate varies between 3.758-14.2 mg/l with a mean of 8.97 mg/l. Fluoride concentration varies between 0.78-1.65 mg/L with a mean of 1.215 mg/L where only 2 samples exceed WHO limit (1.5 mg/L). The water quality index (WQI) is found to be87.32 which suggests that the groundwater is very poor and only suitable for irrigation. It is concluded that about half of the samples failed to meet the water quality criteria set by WHO for drinking. The problem is aggravated due to anthropogenic activities mainly by sewage infiltration and water supply conditions. © International Journal of Ground Sediment & Water, © Sun Jichao, The website is http://ijgsw.comze.com/

Summary Cystic echinococcosis (CE) is a zoonotic disease caused by the larval stage of Echinococcus granulosus tapeworms. These parasites have a worldwide geographic distribution and pose a serious threat to livestock industry as well as human health in the endemic areas. CE is widely distributed in Pakistan. However, very few reports are available related to the regional transmission of E. granulosus. A retrospective analysis was conducted of surgically confirmed CE patients who were treated at Shoukat Khanum Memorial Cancer Hospital and Research Centre in Lahore, Punjab Province, Pakistan from 2007 – 2018. In total, 536 CE patients were evaluated during the study period. Cases originated from the provinces of Khyber Pakhtunkhwa (n=336), Punjab (n=147), Baluchistan (n=18), Sindh (n=3), Islamabad (n=2), Gilgit Baltistan (n=1), and Azad Jammu and Kashmir (n=1). An additional 28 cases were from Afghanistan. The highest number of CE cases was reported in 2013 (n=90). Females made up a larger proportion of cases (n=310; 57.8 %) than males (n=226; 42.2 %). Most patients were members of the Pashtun (n=197; 36.7 %), Hindku (n=142; 26.5 %), and Punjabi (n=118; 22.0 %) ethnic groups. The largest number of cysts was obtained from the liver (137/536; 25.6 %). This study showed that CE is likely present throughout Pakistan. In order to control the disease, a comprehensive control program and regional surveillance are needed.

Present study is aimed to calculate the water quality index (WQI) of Surjani Town by using weighted arithmetic index method to assess its suitability for drinking purpose. For calculating WQI, 12 parameters (pH, TDS, EC, Hardness, Na, Ca, K, Mg, HCO3, SO4, NO3 and Cl) have been taken into account. Except pH, all physicochemical parameters are exhibiting the maximum prominence in WQI quality rating scale (Qn) which suggests that groundwater is highly polluted. The computed value of water quality index (WQI= 331.62) is also found to be high which is comparable with enormously high concentrations of physicochemical parameters that are violating the WHO admissible limit for drinking purpose. Results revealed that the groundwater is under the influence of anthropogenic activity from nearby Jam Chakro solid waste dumping site. It is concluded that groundwater is highly deteriorated which is immensely inappropriate for drinking purpose according to WQI rating score.

In wireless communication many packets may be lost during handoff. In conventional wired networks Transmission Control Protocol (TCP) is used. But in case of wireless communication TCP needs to be modifies according to the environment because if a packet drops during communication it reduces the congestion window sizes excessively due to congestion. To overcome this problem many extension in TCP has been proposed. The purpose of this research is to analyze the performance of TCP and Freeze -TCP in the wireless environment. The simulation was done using Network Simulator-2 (NS-2) for both the TCP and Freeze-TCP for managing the mobility and handoffs and the result was compared according to the performance. The simulation is categorized in to two scenarios Varying Window Size, Varying Segment Size on the basis of window and segment sizes. Each scenario consist of three window sizes and three segment sizes. For each and every scenario the performance of TCP and Freeze-TCP are analyzed and compared. The result shows that for managing the handoffs in mobile wireless communication the F-TCP performs better than that of simple TCP.

In this paper modified opportunistic routing (MORR) protocol is propose, which focuses on addressing routing issues for wireless sensor networks (WSNs) for prolonging network lifetime and load balancing by selecting forwarder nodes, which have maximum residual energy so that network lifetime is enhanced. Furthermore, throughout the selection of forwarder node, channel interference is also taken into account to offer consistent communication. Consequently proposed scheme minimizes the chances of collision and packet duplication at the network layer by controlling number of forwarders and transmitters. Simulation results show that our scheme outperforms in term of average number of transmissions per packet, packet delay, energy consumption and residual energy as compared to baseline solution.

Underwater Wireless Sensor Networks (UWSNs) have been considered as an emerging and promising method for exploring and monitoring deep ocean. The UWSNs face many challenges such as noise, high transmission delays, high deployment cost, movement of nodes, energy constraints, etc. In UWSNs nodes are sparsely and unevenly deployed, that may results in void hole occurrence. Secondly low propagation speed in UWSNs causes high end-to-end delay and energy consumption. In this paper, we propose two schemes: Adaptive Transmission Range in WDFAD-DBR (ATR-WDFAD-DBR) and Cluster Based WDFAD-DBR (CBWDFAD-DBR). In aforesaid scheme to reduce the probability of void hole this scheme adjusts its transmission range when it finds a void node and then continues to forward data towards the sink. In later, to minimize end-to-end delay and energy consumption network is divided into clusters. Simulation results show that our schemes outperform compared with baseline solution in terms of average Packet Delivery Ration (PDR), average energy tax, end-to-end delay and Accumulated Propagation Distance (APD).

Introduction: Little is known about the role of P2Y12 oligomerization in receptor function and whether P2Y12 receptor mutations associated with human bleeding disorders may be explained by alterations in oligomerization. Objectives: 1) To determine whether P2Y12 homo- and hetero-oligomers are constitutive or dynamically regulated. 2) To explore whether P2Y12 mutants R256Q and R265W (previously detected in patients with abnormal bleeding, but with unaltered ADP binding) have different oligomerization affinities or kinetics and determine whether differences in P2Y12 oligomerization explain the functional defects. Methods: We employed a Venus-based Bimolecular Fluorescence Complementation (BiFC) approach in HEK293T cells transiently co-expressing P2Y12 or its mutant forms (R256Q or R265W) tagged with either the N-terminal (P2Y12-VN) or C terminal fragment (P2Y12-VC) of Venus, to characterize their homomeric interactions, in live cells using confocal microscopy and quantitative flow cytometry assays. Results: Agonist-independent formation of P2Y12 receptor homo-oligomers were detected on cell membranes. Time lapse imaging showed movement of P2Y12 receptor pairs from the endoplamic reticulum and Golgi network to the plasma membrane, suggesting that they are constitutive and required for export. Co-expression of P2Y12-VN with increasing amounts of P2Y12-VC demonstrated a dose-dependent increase in the fluorescence intensity of Venus, and reached saturation at a ratio of 1:3. Interestingly, the fluorescence intensities of homomeric P2Y12-R256Q-VN and R256Q-VC and, separately, P2Y12-R265W-VN and P2Y12R265W-VC were almost 4 times stronger than that of the wild type receptor as quantified in flow cytometry-based BiFC. Similar results were obtained in confocal microscopy. This suggests that these P2Y12 mutants form an increased number of dimers or oligomers with increased self-affinities. Conclusion: We demonstrate that P2Y12 forms constitutive homo-oligomers. Two mutations associated with bleeding disorders in patients have altered receptor-receptor interactions. Future investigation will explore the effect of mutations and receptor oligomers on G protein coupling and receptor: G protein stoichiometry.

It is now well accepted that protease activated receptor (PAR) 1 and PAR4 have differential roles in platelet activation. PAR4, a low-affinity thrombin receptor in human platelets, participates in sustained platelet activation in a P2Y12-dependent manner; however, the mechanisms are not defined. Our previous studies demonstrated that thrombin induces the association of PAR4 with P2Y12, together with arrestin recruitment to the complex. Here we show that PAR4 and P2Y12 directly interact to coregulate Akt signaling after PAR4 activation. We observed direct and specific interaction of P2Y12 with PAR4 but not PAR1 by bioluminescent resonance energy transfer when the receptors were coexpressed in human embryonic kidney 293T cells. PAR4-P2Y12 dimerization was promoted by PAR4-AP and inhibited by P2Y12 antagonist. By using sequence comparison of the transmembrane domains of PAR1 and PAR4, we designed a mutant form of PAR4, "PAR4SFT," by replacing LGL194–196 at the base of transmembrane domain 4 with the corresponding aligned PAR1 residues SFT 220–222. PAR4SFT supported only 8.74% of PAR4-P2Y12 interaction, abolishing P2Y12-dependent arrestin recruitment to PAR4 and Akt activation. Nonetheless, PAR4SFT still supported homodimerization with PAR4. PAR4SFT failed to induce a calcium flux when expressed independently; however, coexpression of increasing concentrations of PAR4SFT, together with PAR4 potentiated PAR4-mediated calcium flux, suggested that PAR4 act as homodimers to signal to Gq-coupled calcium responses. In conclusion, PAR4 LGL (194–196) governs agonist-dependent association of PAR4 with P2Y12 and contributes to Gq-coupled calcium responses. PAR4-P2Y12 association supports arrestin-mediated sustained signaling to Akt. Hence, PAR4-P2Y12 dimerization is likely to be important for the PAR4-P2Y12 dependent stabilization of platelet thrombi.

Energy conservation is one of the most important factors in Wireless Sensor Networks (WSNs) for reliability since nodes have limited resources of energy. There is a need to design such routing protocols, which efficiently use available energy and prolong network lifetime and stability period. We implement sink mobility in Clusterless Stable Election Protocol (CL-SEP) and proposed Tunnel Routing Protocol (TRP) for WSNs, which is two level heterogeneous. From the simulation results, it is seen that the proposed protocol outperforms the conventional SEP in stability period, network lifetime and throughput. It efficiently utilizes the available energy of the network by using Moving Sink (MS) and prolongs network lifetime and stability period.

Sink Mobility is becoming popular due to excellent load balancing between nodes and ultimately resulting in prolonged network lifetime and throughput. A major challenge is to provide reliable and energy-efficient operations are to be taken into consideration for differentmobility patterns of sink. Aim of this paper is lifetime maximization of Delay TolerantWireless Sensor Networks (WSNs) through the manipulation of Mobile Sink (MS) on different trajectories. We propose Square Routing Protocol with MS (SRP-MS) based on existing SEP (Stable Election Protocol) by making it Cluster Less (CL) and introducing sink mobility., 26th IEEE Canadian Conference on Electrical and Computer Engineering (CCECE2013), Regina, Saskatchewan, Canada, 2013

Transmembrane (TMEM)16f is member of the TMEM16 family of ion channels, recently shown to be essential for optimal Ca2+-dependent phospholipid scrambling in platelets, platelet-dependent pro-coagulant activity and thrombosis. However, relatively little is known about the effect of TMEM16f on platelet signaling, functional activity, and microparticle formation. TMEM16f-/- mice were obtained from Andrea Vortkamp and the platelets of these mice were isolated, stimulated with either dual agonist (thrombin + convulxin) or calcium ionophore (A23187), and platelet surface exposure of phosphatidylserine (PS) and microparticle generation were assessed by both flow cytometry and high resolution immunofluorescent confocal microscopy. To measure PS exposure, annexin V binding to platelet surfaces was quantified: In response to thrombin and convulxin, only 10.65% ± 1.35% of TMEM16f-/- platelets exposed PS after dual agonist exposure, compared to 16.1% ± 2.3%PS+ WT platelets when analyzed by high resolution microscopy. When analyzed by flow cytometry, dual agonist exposure of WT platelets yielded 52.4% ± 7.0% of maximal annexin V binding achieved by 10μM A23187; in contrast, there was no significant increase in annexin V binding detected in TMEM16f-/- platelets treated with dual agonist. Responses to calcium ionophore were also reduced in TMEM16f-/- platelets relative to WT (17.3% ± 8.3% PS+ TMEM16f-/- platelets in response to 1μM A23187 compared to 40% ± 1.4% PS+ WT platelets, by high resolution microscopy). Microparticle generation from TMEM16f-/- platelets compared to WT platelets was also evaluated by high resolution immunofluorescent microscopy and flow cytometry. On average, approximately 5.7 ± 0.33 microparticles were generated by untreated WT platelets however thrombin and convulxin treated platelets yielded 10.66 ± 0.92 microparticles per platelet whereas no significant increase in microparticle generation was observed in TMEM16f-/- platelets after dual agonist treatment. These data suggest that TMEM16f is required not only for PS exposure on platelet membranes, but also for the shedding of PS+ microparticles. TMEM16f-/- mice also have a significant defect in stable occlusive thrombus formation following 10% ferric chloride injury of the carotid artery (0/6 TMEM16f-/- mice form stable thrombi compared to 5/6 WT mice forming stable thrombi under the same conditions). TMEM16f-/- platelets had a slight (15%) reduction in total fibrinogen binding stimulated by PAR4 peptide agonist compared to WT, but showed no significant differences in aggregation to PAR4 agonist peptide or ADP compared to WT control platelets, suggesting that the defect in thrombus formation is likely due to PS- or microparticle-dependent procoagulant activity. This hypothesis is supported by preliminary results demonstrating that when microparticles isolated from ionophore-stimulated WT platelets were injected into TMEM16f knockout mice, 3/3 TMEM16f-/- mice formed stable thrombi, compared to 5/6 WT mice injected with vehicle control. In conclusion, TMEM16f-/- mice are deficient in platelet PS exposure, platelet-derived microparticle formation, and injury-induced thrombus formation, and platelet-derived microparticles appear to contribute to the defect in thrombus formation. Disclosures: No relevant conflicts of interest to declare.

Abstract 3253 We have previously shown that the PAR4 thrombin receptor and the P2Y12 receptor for ADP co-immunoprecipitate from thrombin-stimulated platelets, but not unstimulated platelets. Here we show that PAR4 and P2Y12 directly interact when expressed in HEK293T cells using Bioluminescence Resonance Energy Transfer (BRET) studies, identify a region in the fourth transmembrane (TM4) domain of PAR4 required for its interaction with P2Y12 and, using a mutant variant of PAR4 in the TM4 domain, demonstrate that PAR4 interaction with P2Y12 is required for arrestin recruitment to PAR4. Luciferase-tagged P2Y12 and YFP-tagged PAR4 were expressed in HEK293T cells and demonstrate specific, saturable interaction by BRET analysis of dimer-dependent light emission at 530 nM. PAR4 and P2Y12 can be co-immunoprecipitated from PAR4-stimulated platelets or HEK293T cells stably expressing both receptors, but the receptors fail to co-precipitate from platelets or HEK293T cells pre-treated with apyrase or P2Y12 antagonist. The PAR1 thrombin receptor and the DP1 prostanoid receptor fail to interact with P2Y12 when evaluated by saturation BRET in HEK293T cells expressing the receptors or by co-immunoprecipitation studies in human platelets. Untagged PAR4 was able to compete with YFP-tagged PAR4 for interaction with luciferase-tagged P2Y12, whereas PAR1 was not. Based upon sequence comparison of PAR1 and PAR4 in the transmembrane domains, the most frequently identified sites of G protein-coupled receptor (GPCR) associations, we have identified 3 amino acids at the base TM4 of PAR4 (LGL194-196) that differ significantly from the SFT(220–222) expressed at the aligned sequence of TM4 in PAR1. Mutation of residues LGL194-196 of PAR4 to SFT results in a mutant variant of PAR4 (PAR4-TM4m) that, when expressed in HEK293T cells, supports only 8.8% of the interaction with P2Y12 compared to the native PAR4 according to saturation BRET (average of n=3). However, homo-dimerization of PAR4 does not require this determinant, as luciferase-tagged PAR4-TM4m mutant supports saturation BRET with YFP-tagged PAR4. The PAR4-TM4m variant expressed in HEK293T cells also fails to co-immunoprecipitate with P2Y12 and with arrestin-2, whereas non-mutant PAR4 supports both of these interactions. This is the first demonstration that direct heterodimeric interaction of two GPCRs expressed in platelets is required to recruit the molecular scaffold arrestin-2. Based upon our previous results demonstrating that arrestin-2 is required to recruit PI3K to PAR4 and for sustained, maximal activation of Akt, we hypothesize that the co-recruitment of P2Y12 and arrestin-2 to activated PAR4 in platelets is required for sustaining PI3K-dependent responses to both Rap1 and Akt activation with potential consequences for maintaining platelet plug stability. Disclosures: No relevant conflicts of interest to declare.