CRISPR/CAS9 as a Gene Editing Tool to Delete the Transcriptional Repressor Inducible cAMP Early Repressor from the Zebrafish Genome

Genome editing has become an important tool in identifying the specific function and role of a gene in an organism. With the advent of genome editing using clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated (Cas) systems, editing a gene has become much easier and less expensive. This CRISPR/Cas9 system uses a small 20 nucleotide long guided RNA (gRNA), which along with Cas9 will bind to the target site and cleave it. In this research, CRISPR/Cas9 system was used to knock out the inducible cAMP early repressor (ICER) promoter sequence in zebra fish. ICER has anti-proliferative activity and acts as a tumor suppressor. In Ras-induced melanoma, ICER protein is being targeted to degradation. Knocking out the ICER gene will help us to establish the tumorigenicity of ICER in melanomas. A gRNA, specific towards the ICER promoter sequence was designed in an attempt to cleave it with Cas9. Plasmids pDR274(-)atgICER and pMLM3613 were used to generate gRNA and Cas9mRNA via in vitro transcriptions. To check the efficiency of designed gRNA in vitro, PAC-2 cell lines were transfected with a plasmid that expressed both gRNA and Cas9. The results demonstrate the ability of Cas9 to cleave the target sequence. In the future, this plasmid could be used to perform microinjection in zebra fish embryos with the generated gRNA and Cas9mRNA

Neuro-Protective Effect of Ethanolic Extract of Trombidium Grandissimum Koch Against Aluminium Chloride (Alcl3) Induced Neurotoxicity in Albino Rats

The current goal of this investigation is to assess the ethanolic extract of T. grandissimum (EETG) against AlCl3-initiated neurotoxicity, where AlCl3 (100 mg/kg/p.o) is administered for 42 days by provoking oxidative stress followed by neurotoxicity. The evaluation of behavioural parameters such as locomotor activity, elevated plus maze, and motor incoordination, as well as biochemical parameters such as reduced glutathione, lipid peroxidation, Superoxide dismutase (SOD), Catalase (CAT), total Protein, and Acetylcholinesterase in the brain, was performed. According to the study report, behavioural changes such as dropped locomotor movements, enhanced transfer latency, and muscle weakness were often noted. Significant rising of lipid peroxides and AChE activity in rat brain tissue was observed, and activity of antioxidant enzymes like SOD, Catalase, and Reduced Glutathione decreased substantially in alcl3 treated rats compared to control, whereas EETG at both dose and standard treated rats showed a significant reduction in LPO, AChE activity and significant elevation in SOD, CAT, and GSH in brain tissues compared to AlCl3 treated rats. The findings are further supported by histopathology of the brain. This is due to the extract's enormous amount of bioactive components. Through in-silico analysis, isolation, and characterisation, future research will be required to pinpoint the chemical component responsible for the extract's observed biological effect

An Optimized Congestion Control and Error Management System for OCCEM

In this paper, a new OCCEM congestion control mechanism is introduced in multi-homing mode. Congestion in each route can be avoided or can be controlled, based on an Active Queue Management (AQM) method. Also, routers compute probability of congestion for the sources on the paths and then notify them. Therefore, the sources can adjust their sending rates on each path effectively and if necessary, can switch to an alternate path to prevent congestion. Simulations have been conducted with Opnet linked with NS2. The simulation shows that the new method can decrease packet loss, increase the amount of transmissions and stabilize queue length, as compared with standard OCCEM. Keywords: OCCEM; AQM; congestion contro

Routing and Broadcast Development for Minimizing Transmission Interruption in Multi rate Wireless Mesh Networks using Directional Antennas

Using directional antennas to reduce interference and improve throughput in multi hop wireless networks has attracted much attention from the research community in recent years. In this paper, we consider the issue of minimum delay broadcast in multi rate wireless mesh networks using directional antennas. We are given a set of mesh routers equipped with directional antennas, one of which is the gateway node and the source of the broadcast. Our objective is to minimize the total transmission delay for all the other nodes to receive a broadcast packet from the source, by determining the set of relay nodes and computing the number and orientations of beams formed by each relay node. We propose a heuristic solution with two steps. Firstly, we construct a broadcast routing tree by defining a new routing metric to select the relay nodes and compute the optimal antenna beams for each relay node. Then, we use a greedy method to make scheduling of concurrent transmissions without causing beam interference. Extensive simulations have demonstrated that our proposed method can reduce the broadcast delay significantly compared with the methods using omnidirectional antennas and single-rate transmission. In addition, the results also show that our method performs better than the method with fixed antenna beams. Keywords: Multihop, Wireless, Mesh, Omnidirectional 

Hop-by-hop Channel - Alert Routing to Congestion Control in Wireless Sensor Networks

One of the major challenges in wireless sensor networks (WSNs) research is to prevent traffic congestion without compromising with the energy of the sensor nodes. Network congestion leads to packet loss, throughput impairment, and energy waste. To address this issue in this paper, a distributed traffic-aware routing scheme with a capacity of adjusting the data transmission rate of nodes is proposed for multi-sink wireless sensor networks that effectively distribute traffic from the source to sink nodes. Our algorithm is designed through constructing a hybrid virtual gradient field using depth and normalized traffic loading to routing and providing a balance between optimal paths and possible congestion on routes toward those sinks. The simulation results indicate that the proposed solution can improve the utilization of network resources, reduce unnecessary packet retransmission, and significantly improve the performance of WSNs. Keywords: Wireless sensor networks; Traffic-aware; Routing; Data transmission rate; Congestion; Gradien

Survey on Quality Analysis of Collaboration Motivation Approach in Wireless Ad Hoc Networks

In Wireless Ad Hoc Networks (WANETs), tasks are conducted based on the cooperation of nodes in the networks. However, since the nodes are usually constrained by limited computation resources, selfish nodes may refuse to be cooperative. Reputation system is one of the main solutions to the node non cooperation problem. A reputation system evaluates node behaviours by reputation values and uses a reputation threshold to distinguish trustworthy nodes and untrustworthy nodes. Although this system has been widely used, very little research has been devoted to investigating the effectiveness of the node cooperation incentives provided by the systems. We propose a protocol called Enhanced Reverse Ad Hoc On Demand Vector Routing Protocol (ERAODV), which uses Hybrid Reputation System (HRS). A Hybrid Reputation system is an enhanced version of Classical Reputation System (CRS). Unlike the CRS it takes into account all the reputation values from the node to determine whether it is trustworthy or not. Keywords: WANET; Reputation System; Price Based System; Quality Analysis; Multipath Routing

Sector Routing Protocol (SRP) in Ad-hoc Networks

Routing protocols for wireless ad-hoc networks face the challenge of dynamic topology due to node mobility, limited channel bandwidth and low transmission power. Both proactive and reactive protocols have trade-off in them. Proactive protocols have large overhead and less latency while reactive protocols have less overhead and more latency. The ZRP is a hybrid protocol that overcomes the shortcomings of both proactive and reactive routing protocol. ZRP divides the entire network into overlapping zones of variable size where routing inside the zone is performed using proactive approach and outside the zone is performed using reactive approach. Keywords: Ad-hoc Networks, Routing, Reactive, Proactive, ZRP

Antifungal activity of triterpenoid isolated from Azima tetracantha leaves.

The present study was designed to evaluate the antifungal activity of Azima tetracantha extracts and isolated compound (friedelin) against fungi. Antifungal activity was carried out using broth microdilution method and fractions were collected using (silica gel) column chromatography. The antifungal activity of Azima tetracantha crude extracts and isolated compound (friedelin) were evaluated using the micro dilution method. Hexane extract showed some antifungal activity. The compound also exhibited antifungal activity against tested fungi. The lowest MIC against Trichophyton rubrum (296) was 62.5 microg/ml and the MIC for Curvularia lunata was 62.5 microg/ml. These results suggest that Friedelin is a promising antifungal agent

Possible role of Toxoplasma gondii in brain cancer through modulation of host microRNAs

Background: The obligate intracellular protozoan parasite Toxoplasma gondii infects humans and other warm-blooded animals and establishes a chronic infection in the central nervous system after invasion. Studies showing a positive correlation between anti-Toxoplasma antibodies and incidences of brain cancer have led to the notion that Toxoplasma infections increase the risk of brain cancer. However, molecular events involved in Toxoplasma induced brain cancers are not well understood. Presentation of the hypothesis Toxoplasma gains control of host cell functions including proliferation and apoptosis by channelizing parasite proteins into the cell cytoplasm and some of the proteins are targeted to the host nucleus. Recent studies have shown that Toxoplasma is capable of manipulating host micro RNAs (miRNAs), which play a central role in post-transcriptional regulation of gene expression. Therefore, we hypothesize that Toxoplasma promotes brain carcinogenesis by altering the host miRNAome using parasitic proteins and/or miRNAs. Testing the hypothesis The miRNA expression profiles of brain cancer specimens obtained from patients infected with Toxoplasma could be analyzed and compared with that of normal tissues as well as brain cancer tissues from Toxoplasma uninfected individuals to identify dysregulated miRNAs in Toxoplasma-driven brain cancer cells. Identified miRNAs will be further confirmed by studying cancer related miRNA profiles of the different types of brain cells before and after Toxoplasma infection using cell lines and experimental animals. Expected outcome The miRNAs specifically associated with brain cancers that are caused by Toxoplasma infection will be identified. Implications of the hypothesis Toxoplasma infection may promote initiation and progression of cancer by modifying the miRNAome in brain cells. If this hypothesis is true, the outcome of this research would lead to the development of novel biomarkers and therapeutic tools against Toxoplasma driven brain cancers

Developement of Evolution Algorithm for Shop Scheduling problem

Tato diplomová práce je zaměřena na výzkum evolučních algoritmů (EA) v oblasti plánování zakázkové výroby a na vývoj nové strategie za účelem zlepšení výkonu. Sekvenční rovrhovací problem (JSSP) je jedním z nejsložitějších plánovacích problémů a nalezení optimálního řešení je vzhledem ke složitosti velmi obtížné. Byly přezkoumány existující evoluční algoritmy a pro řešení sekvenčního rozvrhovacího problému byl vybrán jeden z široce používaných genetických algoritmů. Pro porovnání efektivnosti EA jsou vygenerovány nejprve Aktivní plány pro pro eta lonové problémy JSSP na základě různých prioritních pravidel . Poté je přezkoumána struktura a hlavní parametry jednoduchého genetického algoritmu (SGA) a na základě toho je v SGA navržena a implementována nová strategie nahrazení (opakovaně použitelná substituční strategie - RSS). Implementace RRS v SGA zlepšuje výsledky a také byl experimentován její dopad na dva různé typy reprezentací chromozomů. Navržený MSGAJO je považován mezi testovanými za nejlepší genetický algoritmus, který dává nejlepší hodnoty promísení pro případy problému JSSP.This thesis is aimed at research of evolution algorithms (EA) in the field of the shop scheduling problems and to develop a new strategy in order to improve the performance. Job shop scheduling problem (JSSP) is one of the most complex scheduling problem and finding the optimal solution is very difficult due to their complexity. Existing evolution algorithms were reviewed and one of the best and widely used genetic algorithm is selected for solving job shop scheduling problem. Active schedules for JSSP were generated based on various dispatching rules with the help of most used problem instances to compare effectiveness of EA. Then the structure and the major parameters of simple genetic algorithm (SGA) is reviewed and based on that a new strategy for replacement (Reusable Replacement Strategy) is proposed and implemented in the SGA. The implementation of RRS in SGA improves the results and also its impact on two different type of chromosome representations were experimented. The developed MSGAJO is concluded to be the best genetic algorithm among tested to give the best makespan values for the JSSP problem instances