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FireEye's and Cisco's endpoint protection solutions both made eSecurity Planet's record of dependable endpoint detection and response (EDR) solutions, and every product offers massive benefits to industry users. What follows is a glance at every solution's key facets and up to date enhancements, together with an analysis of their strengths and weaknesses.
both options possess enthusiastic supporters, with the immense majority of users epigram they'd insinuate both products to others. Key strengths of FireEye's offering involve the probability intelligence it receives from the business's Mandiant subsidiary, as neatly as the solution's new MalwareGuard detection and prevention engine. FireEye's managed detection and response provider is one key differentiator for valued clientele brief on resources. Cisco's cloud-managed admit merits from Talos possibility intelligence, and a lot of users cite the providing's ease of deployment as a key energy.
Overview: Cisco AMP for Endpoints (advanced Malware insurance device for Endpoints) is a cloud-managed solution designed to avoid, become aware of and reply – combating breaches and blocking off malware at the aspect of entry, then detecting, containing and remediating advanced threats. The admit immediately correlates data, telemetry statistics, deportment and pastime to simplify investigations and abridge mitigation time, and the AMP Cloud is consistently up to date with tips from Cisco Talos and risk Grid to provide real-time danger intelligence.
contemporary tendencies: fresh enhancements to AMP for Endpoints consist of an stronger machine Trajectory characteristic that suggests a extra in-depth view of the endpoint and helps users find spikes in endpoint activity. a brand new hazard severity characteristic offers activities severity tags of critical, high, Medium or Low. a new Overview page offers a quick overview of a company's endpoint protection state, and the Casebook built-in case management device assists in gathering and pivoting on observables, assigning names to investigations, taking notes, and extra.
Analysts' take: Gartner says the leading strengths of Cisco AMP prevaricate in random intelligence and hold handicap of prevention, with step by step enhancing integration and data sharing between AMP and other Cisco security choices such as networks, firewalls, and many others. still, the analysis hard says that whereas the statistics it gives is relatively comprehensive, the admit requires distinct clicks across distinct monitors to pick up a full figuring out of the condition of an endpoint or of the considerations being led to by way of malware.
Overview: FireEye Endpoint security makes exhaust of a signature-based endpoint protection platform (EPP) engine to dam regular malware, MalwareGuard computer studying to locate threats for which a signature doesn't yet exist, a habits-based analytics engine to allow EDR capabilities against superior threats, and a true-time indicators of compromise (IOC) engine to locate hidden threats.
contemporary tendencies: fresh additions to the providing consist of a signature-based prevention engine to clear out prevalent malware, viruses and worms; and the desktop studying-based mostly MalwareGuard to give protection to from prior to now unknown threats. The MalwareGuard desktop getting to know mannequin is informed with each inner most and public statistics sources, together with data gathered from greater than 15 million endpoint agents, beset analyses in keeping with over a million hours spent responding to assaults, more than 200,000 consulting hours every year, and adversarial intelligence gathered from a worldwide community of analysts speaking 32 languages.
Analysts' take: Gartner studies that FireEye's offering merits from risk intelligence from Mandiant's infringement investigation crew and iSIGHT probability Intelligence provider in addition to from FireEye products' shared hazard indicators, and that its managed detection and response provider is alluring to consumers which are brief on resources. still, the analysis hard says some purchasers file that the solution produces excessive erroneous high property quotes when first carried out.
listed here are eSecurity Planet's scores of every answer's key aspects.
consumers of each vendors document tough efficiency. essentially the most fresh Forrester Wave record on EDR options gave FireEye a rating of 3.08 out of five and Cisco a rating of 2.84 out of 5. The rating is in keeping with various criteria including configurability, agent effectiveness, forensic capabilities, deployment alternatives and response movements.
In fresh trying out, Forrester rated FireEye's detection capabilities at three.0 out of five, and its response capabilities at three.four out of five. Cisco's detection capabilities obtained a score of 1.eight out of five, and its response capabilities 2.2 out of five. Cisco additionally bought a informed score and ninety four.5% safety effectiveness rating from NSS Labs. Any NSS consequences from FireEye possess yet to breathe launched.
consumers of both Cisco and FireEye often document satisfaction with pricing and value for money, even though in both circumstances some discipline the solution is extra costly than others.
FireEye clients record exceedingly smooth deployments. a number of Cisco reviewers peculiarly noted the installation manner as a key energy, asserting it became primary and straightforward.
FireEye users file nice experiences with customer support. Some Cisco reviewers expressed frustration with the level of aid received.
both options present both cloud-primarily based and on-premises deployment options.
Gartner Peer Insights clients gave FireEye four.4 stars out of 5 and Cisco four.2 stars out of five, although 82 percent observed they would insinuate FireEye to others, and 88 % spoke of the very of Cisco.
IT relevant Station reviewers were additionally nice on each FireEye and Cisco.
Cisco reviewers observed "implementing AMP turned into elementary," that it "often would not influence performance in a extremely remarkable way," and that "the cloud administration simplifies administration." nevertheless, others said "computer virus fixes and enhancements are tremendous slow," and that early on in the deployment, "the erroneous positives possess been a bit irritating."
FireEye reviewers noted the admit "gives a superb toolkit for monitoring, detecting and responding to threats on the endpoint stage across the commercial enterprise," and that "assist is very good." still, others said the admit is "on the high priced facet," and has brought about some "difficulties with firmware updates and habitual product bugs."
Cisco AMP for Endpoints supports cloud and on-premises deployments. agents are available for home windows, Mac and Linux, in addition to Android and iOS.
FireEye Endpoint protection supports cloud, on-premises and hybrid deployments. agents can breathe found for windows, Mac and Linux.
Cisco AMP for Endpoints is licensed on a one, three or 5-yr subscription time period, with pricing in response to the variety of endpoints covered. The longer the subscription term and the extra endpoints covered, the lower the freight per user.
FireEye Endpoint protection is purchased through a subscription mannequin in line with the stage of insurance policy and investigation apparatus available – the elementary edition begins at $39 per endpoint, and the more advanced vigour version begins at $fifty eight.50 per endpoint, with volume discounts obtainable for both. Free trials are available.
The latest edition of Cisco integrated Communications system (ICS) 7750, an IP PBX offering targeted at groups or industry offshoot places of work with up to 1000 employees, now supports Cisco's solidarity Unified Messaging four.0, referred to Bruce Laird, vp and Popular manager of the built-in Communications neighborhood at Cisco. solidarity four.0 permits users to manage voice mail, email and faxes from a solitary machine. up to now, ICS 7750 supported simplest the voice-mail function of the team spirit utility, Laird stated.
ICS 7750 now additionally supports Cisco client Response admit (CRS) 3.0, Laird said. including CRS capabilities to ICS 7750 permits midsize agencies and industry department workplaces to hasten "relatively refined" cognomen centers with as much as 50 brokers, he spoke of.
messaging platform and compose contact with middle application.>>%A0IP telephony content material has been added to e-learning software and web enterprise Roadmap sales application.>>%A0Special rent kit extends terms to five years and contains hardware, software and capabilities.
Cisco furthermore delivered reduced-price admit bundles along with the up to date points. every package comprises the newest version of Cisco CallManager, Cisco solidarity Voice Messaging and Cisco Auto Attendant. The packages may furthermore breathe upgraded to back as much as 500 clients.
The entry-degree expense for ICS 7750 voice packages, supporting up to 50 users, is now $14,995, Laird mentioned. For businesses with centralized cognomen management, Cisco additionally unveiled new voice-package pricing for its offshoot routers. With the programs, Cisco 1700 and 2600 collection router platforms can serve branch-workplace IP telephony administered from a company's headquarters.
For channel partners, Cisco introduced ICS 7750 classes to its e-discovering portfolio, Laird pointed out.
additionally, the networking hardware vendor is adding ICS solutions content to the Cisco web enterprise Roadmap, an internet-based mostly solutions-selling software for Cisco partners, he talked about.
Slated to breathe posted by means of mid-February, the brand new IP telephony materials will consist of network blueprints, case reports, road maps and how-to-sell and why-to-buy displays, Laird noted.
besides the particular pricing and training programs it plans to present companions, Cisco furthermore created a leasing application that is more pleasant to midsize agencies, he talked about.
Cisco methods Capital is now offering a 5-12 months rent on IP telephony solutions, enabling for the hardware, application and functions contracts to breathe covered within the rent plan, he mentioned. as much as 36 months of Cisco's SmartNet provider device will furthermore breathe blanketed in the hire software activity-free, Laird introduced.
Ethan Simmons, a accomplice at Boston-primarily based admit company NetTeks, referred to he mainly appreciates the brand new hire terms.
"Cisco's leases possess been short-term, which is indifferent within the facts world, however businesses hire telephone programs for five to seven years," Simmons referred to. "Any time the lease alternatives are enhanced, it helps sell more methods."
shoppers commonly quiz us whether synthetic intelligence (AI) will breathe the benevolent of online game changer that analysts are predicting. From what I'm seeing, the reply is an unconditional sure.
The trick is determining the appropriate exhaust circumstances. whereas any computer can cipher pi to 1,000,000 locations faster than i can sneeze, it takes an AI compute house to character through billions of items of statistics to reply a solitary query. One drawback: AI doesn't, via itself, recognize what questions to quiz and what to sequel with the answers. but if that you would breathe able to body the query the perquisite means, AI can devour planet numerous suggestions and find massive patterns. believe of it this way: AI can winnow via massive haystacks to find a needle—however first a human needs to define a needle.
Cisco IT whole started their journey to AI through many disconnected teams. in the course of the grapevine I'd heard about greater than 40 initiatives, every funded by means of the group that's using it (e.g., marketing, InfoSec, contact center). diverse groups in Cisco – in IT, in Engineering, in advertising – possess already centralized their AI efforts to expand their scope.
listed here are probably the most methods we're placing AI to work nowadays.
Malware detection turned into their first foray into AI. They bought Stealthwatch technology in 2015. It sifts through billions of facts aspects about how traffic moves via their community to notice anomalous conduct that could point out Day Zero malware. The hard half is teaching the AI engine what's habitual and what's no longer. here's an analogy: in case you possess a cocktail party with thousands and thousands of visitors (it's a large apartment), how are you able to spot the learning thieves? Most protection defenses behold for signatures – during this case, mug-pictures of customary criminals as they Come in the course of the door. however to seize the thieves who are nevertheless unknown, you must behold at their habits. it's commonplace for a party guest to ramble via your house, speaking to visitors and hovering around the bar; but it's now not generic for them to head automatically to the locked latitude containing the secure and commence searching behind the images. When the equal of that happens on their community, Stealthwatch raises an warning or takes action to segregate the probability. They continuously find Day Zero assaults during this way. To sequel that, they possess to quiz Stealthwatch to sift through over 28 billion Netflow statistics day by day, and continue to update it on what's average, suited deportment and what conduct is assign of malware attacks. however makes it practicable for us to espy issues that no other device can.
greater than 50% of network site visitors is now encrypted, and malware hidden in encrypted site visitors can sneak through ordinary defenses. however encrypted site visitors is so challenging to crack (pun meant) that you furthermore want laptop learning—telling the AI application to locate malware devoid of telling it how. the exhaust of an AI software called Encrypted traffic Analytics, a new Stealthwatch upgrade, we've learned some of the clues, akin to packet lengths, arrival instances, and prefatory handshake facts packets that signify malware even when the movement continues to breathe encrypted.. ETA has discovered malware in encrypted streams that might possess slipped appropriate with the aid of signature analysis and even AI-primarily based habits evaluation.
Our midsize workplaces pick up two circuits: MPLS and VPN-over-internet. as a substitute of leaving the secondary circuit idle many of the time, final yr they began the usage of Cisco software defined-WAN (SD-WAN) to intelligently provision relaxed WAN links and route application-certain site visitors to the circuit that's most profitable for the job. The determination depends upon the class of the traffic (voice, video, e-mail, and many others.) and latest community circumstances. (For extra, hold a behold at this weblog through my colleague Carol Goh.) Now we're making the determination even more suitable through the exhaust of AI to predict future circuit conduct. discipline we're a few broadcast a reside 60-minute webinar. If the MPLS circuit is performing wonderful perquisite now however signs point out it might humble in 30 seconds (or 17 minutes), it's smarter to possess the SD-WAN supervisor route site visitors to the backup circuit.
Cisco software-defined access (SD-A) includes an AI-driven statistics collection and analysis platform. Cisco IT has already deployed three main Cisco DNA-core (DNA-C) clusters, one in every of the three global regions (Americas, Europe, Asia). These AI clusters are amassing colossal amounts of suggestions concerning change traffic and efficiency, monitoring traffic from every software and person. (This has required us emigrate a few thousand switches to yeast 9000 fashions, which act as sensors to stream telemetry information to the Cisco DNA-C for analysis.) dote whole AI device, Cisco DNA-C benchmarks ordinary deportment and performance. It identifies when performance is degrading and consults over 100 generic Cisco IT community considerations. If it finds the perquisite pattern, it will alert a community engineer, element out precisely the site in the community route there is a problem, and recommend an admit in accordance with that pattern. The significant Cisco DNA-Controller can then instantly compose advised alterations to whole vital network gadgets.
We've discovered the instant Assurance a fraction of Cisco DNA-C to breathe extraordinarily advantageous. it could stitch together the course of an individual running across the pile floor, connecting from one access aspect to the next, and spot exactly where and when their voice or video session starts to hasten into issues, as well as making a election on where within the customer device, entry aspect or wired network the issue root occasions can be. If it suits probably the most hundred-plus regular considerations, it'll advocate a fix and walk the community engineer through that repair.
AI apparatus akin to the networking tools described for WAN and LAN are additionally at work within the much more advanced environment of the information center. With hundreds of distinct sets of utility efficiency and security guidelines in area, enforced by the digital overlay fabric across ACI, it's now not handy to deploy new utility policies without considerations. Cisco network Automation Engine (CNAE), a brand new AI device, will instantly assess for brand new coverage conflicts among the many millions of distinctive capabilities connections to espy the site issues might Come up, and recommends diverse guidelines to achieve the preferred outcome. This continues software protection and performance at a highest, with minimal provisioning lengthen as a result of misconfiguration anywhere in the facts core. Cisco it's running CNAE in the greatest of their three ACI-material data centers nowadays.
Our small and medium enterprise purchasers often sequel their product research on cisco.com. over the years we've experimented with quite a few methods to keep up with internet friends. e mail or cell keep up isn't primarily effectual and may display to breathe spam.
Now we're the usage of AI to find the next most appropriate action in line with the client's company want and former interactions. Working with their advertising Analytics crew, they developed a platform that collects and analyzes counsel from cisco.com and Salesforce to learn the way valued clientele were contacted, what content material they were given, and whether the motion successfully moved the consumer up the sales chain (for instance, inspiring them to attain out to us, watch a product video, location an order, and so on.) consequently, they now comprehend which valued clientele are more likely to respond to clear sorts of contacts, and at what point in the purchase decision. prefatory effects from 25 pilots in 7 nations are very amazing: four times more suitable customer response price, 7-10 times fewer outbound communications that don't influence in a response, and reduce expenses. Even stronger, we've viewed that the longer they tune the records, the greater the response rates over time.
Self-provider for callers and brokers. Cisco IT worked with the contact heart crew to build Cisco answers, an AI-driven handicap device.
intelligent routing: When customers contact us by way of voice, e-mail, or chat. they exhaust AI to predict what they want, after which associate them to the top of the line attainable agent with the perquisite advantage. First-name resolution and customer delight possess both greater.
business insights from recorded consumer calls. dote loads of groups, they listing contact heart interactions for agent working towards. These recordings are additionally a gold mine of suggestions for advertising, product building, and extra. With near 100,000 of calls/day, a human couldn't sustain up—however AI can. We've started the usage of Verint speech analytics to find traits in their recorded contact middle interactions. as an instance, if they espy a spike within the phrase "application described" in communications with individuals in a inescapable location, they could step up their advertising classes for application defined Networking in that vicinity.
other grassroots AI tasks are cropping up in whole places Cisco—in income, advertising, provide chain, and others. hold give chain. The Cisco server you order these days likely isn't developed yet as a result of they exhaust simply-in-time (JIT) manufacturing. To compose it work, they want the remedy accessories reachable, just after they exigency them. The more accurately they can predict exact a month or two out, the much less risk that we'll under order whatever dote chips, delaying cargo—or over order, tying up capital and growing the risk of loss or damage. It's looking dote AI will aid us reduce stock requirements for americaserver reminiscence chips via a factor of ten.
The exhaust circumstances above are only a sampling of the alternative ways we're leveraging the vigor of AI to automate extremely involved choice-making strategies. they now possess the records. they now possess the compute vigor. What they exigency greater of is people who breathe aware tall statistics, AI, and laptop discovering, and we're at the moment re-skilling and hiring to immediate the IT skill hole.
How are you at present using AI in IT operations, and what are your hopes—purposeful and grand? I invite you to share your recommendations under.
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PHOENIX, Dec. 18, 2018 /PRNewswire/ -- Execs In The Know, a community of senior brand leaders dedicated to customer experience, possess announced the details of their national event – Customer Response apex (CRS) New Orleans. The apex will breathe held February 4th-6th, 2019. Customer engagement, AI, emotional loyalty, advanced technologies, and other CX topics will breathe covered over three days of discussion, case studies, panels, and small group presentations. The conference will involve keynotes from executives at Tesla, HomeAway at Expedia, Inc., and Peloton.
The Customer Response apex brings customer service senior leaders across varied brands and verticals together, with the objective of bringing excellent customer flavor alive in their own organizations. The three-day apex will feature top-tier brands sharing insights, engaging in networking opportunities, and presenting real-world CX solutions.
"I am continually impressed with the individuals that associate us at their Summits," said Chad McDaniel, President and CEO of Execs In The Know. "We attract innovative brand leaders to their community, making it a diverse and dynamic group of people whose main goal is to mosey the needle and achieve the next level of CX greatness. For those who possess not been a fraction of a CRS, I encourage leaders to associate us in New Orleans—there's no event dote this one."
Other leading corporate brands that will talk at the customer keeping event involve Quicken Loans, Waymo, Square, Symantec, Groupon, Marriott, and more.
To learn more, visit the event website at https://www.execsintheknow.com/events/crs-new-orleans/.
Execs In The Know is a global community of customer flavor professionals focused on excellence in customer experience. Execs In The Know gives brands a platform to share and gain insights, benchmark their brand, linger on top of the latest trends in CX, and create lasting relationships with their peers – "Leaders Learning From Leaders."
Execs In The Know holds numerous live events each year including Customer Response Summit, matter Matter Briefings, Lunch & Learns, and Dinners. They furthermore present industry content and thought leadership, webinars, reports, and various sociable communities. Learn more at www.execsintheknow.com.
In the early 90's Microsoft was facing fierce competition in the operating system (OS) market. Included among the gathering storm of competition was a company called Digital Research, which released a product called DR-DOS 6.0 into the operating system space. This system came with some fantastic built in features and allowed users to sequel Interesting things which were considered ahead of the curve at that time.
Microsoft sensed a threat. Customers now had other viable options in the OS space.
As Microsoft weighed whole of their options, one thing they acted on quickly was an organized pains to compose the public nervous about using competitor products. Management employed tactics based on a well-worn sales style called F.U.D. – an approach centered around stoking a sales prospects' Fear, Uncertainty, and Doubt.
They implemented F.U.D. by coding their operating software to parade a "non-fatal error" message to anyone who used DR-DOS on Windows 3.0. The understanding was that users would cerebrate that something was wrong with their system and that doubt would give the Microsoft sales team an edge they could exploit.
Ultimately, Microsoft's less-than-honest approach was exposed and they had to pay just under $300 million in settlement fees to Digital Research.
More modern examples of F.U.D. normally Go dote this: "Buy their software so you don't linger up dote Target and possess 70 million of your users' data hacked." Or, "You can Go with them because their expense is lower, but their customers possess experienced significant data loss." It may breathe effectual in the cyber world, but outside of the computer and data security industries, examples of F.U.D. are often plagued with horror stories dote the one above from Microsoft.
This has given F.U.D. an unnecessarily soiled name. The sales principles espoused within F.U.D. possess merit, and they don't possess to breathe associated with underhanded selling or lying claims.
All that is really needed is a renewed approach to F.U.D. in a way that aligns with the unique sales challenges of 2019 and dispenses with things that erode prospect trust.
When sales pros cerebrate of using scare to their handicap in their outreach efforts, one of the first thoughts to pop up is something akin to smear campaigns in politics. This is misguided. You don't possess to create scare in a prospects' minds by bashing a competitor, making a fake mistake message, or using a slippery slope argument. scare is already present in every solitary prospect.
Commonly, scare shows up in more traditional ways, such as concerns over losing market share or being ultimate to adopt a new technology. If you're sales prospects work for cloud-based storage firms, for example, referencing ways in which you can serve them compete against Amazon Web Services (AWS) — probably one of their top priorities — is going to resonate.
"Hi (Prospect), I noticed you're fighting off a near monopoly in your space from AWS, and LinkedIn shows that your finance team has grown over 12% so far his year alone. I'd keeping for to introduce you to how they are helping finance departments dote yours automate and consolidate the approvals process. The time saved could breathe used to fight back against the evil empire." (pop culture reference not needed).
The prospects' scare of competing with the market leader are not played on here, they are simply acknowledged. Crafting messages in this way avoids the unethical and nefarious parts of F.U.D., while offering an approach made for prospects in 2019.
Another point worth emphasizing perquisite here – you're going to possess to work to personalize prospect pain. The cloud-storage company in the illustration above could breathe going after different verticals than AWS and their true top competitor is HP's agile or PureStorage.
A recent study that analyzed data from over 100,000 sales prospecting chilly calls showed that referencing companies who you work with that are not similar in size to the prospects' company leads to a 22% lower immediate rate. Referencing Microsoft as a customer may compose startups feel dote you target a different character of company. Imagine what referencing the wrong throe points or misguided (assumed) concerns to a sales prospect will sequel to your immediate rates.
Sales is essentially the transference of certainty. In sales development efforts, it's sensible to assume that every prospect you talk with on the phone is uncertain of whether they want to meet with you. In fact, they're often uncertain as to whether they want to even give you 30 seconds to talk before… CLICK.
Your opening on a chilly sales call is crucial.
Within the first 10-20 seconds, sales development pros must establish rapport and quickly mosey to parade value. One effectual way to sequel this is by first asking if a prospect has a moment to talk. By asking this in a inescapable way, using a consultative tone, you're showing you respect their time and admit that you're calling them out of the blue. When they give you the "OK", you're now clear to initiate a discussion with them without getting immediately shut down.
At this stage, your best mosey is to establish credibility. This can furthermore breathe seen as the dissolution of doubt. Your prospect may not possess heard of your company and they're uncertain as to whether or not you can serve them in the least. Mentioning some well-known names you're working with perquisite away, before going into a discussion of your products and services, can Go a long way towards pile credibility.
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SAN JOSE, CA--(Marketwired - Jul 28, 2014) - Cypress Semiconductor Corp. ( NASDAQ : CY ) today announced that Raytrix GmbH has selected Cypress's EZ-USB® CX3™ USB 3.0 camera controller for its new 3-D camera. The EZ-USB CX3 controller, which is now in full production, delivers 5-Gbps performance to the innovative R42 3-D Light bailiwick Camera. The R42 supports up to 40-Megapixel resolution and 4,000 images, leveraging the CX3 solution's throughput to provide 3-D images with extended depth-of-field. The all-around camera is small enough for convenient exhaust in a variety of applications, including machine vision, motion tracking and surface inspection.
EZ-USB CX3 is the industry's most supple USB 3.0 camera controller. It allows developers to add USB 3.0 connectivity to any image sensors supporting the Mobile Industry Processor Interface (MIPI) Camera Serial Interface character 2 (CSI-2) standard. CX3 is ideally suited for high-definition (HD), high-speed image-capturing applications and is capable of streaming uncompressed video at 1080p at 30 frames per second or 720p at 60 fps. The CX3 solution is supported by the new Denebola reference design kit from Cypress's partner, e-con Systems. The kit accelerates time to market for HD cameras with a ready-for-production reference board that leverages CX3's SuperSpeed USB performance to seamlessly process large image files and stream HD video without compression, which degrades image quality.
"The EZ-USB CX3 USB 3.0 controller delivered exactly what they needed for the R42 camera," said Dr. Lennart Wietzke, founder of Ratrix GmbH. "CX3's SuperSpeed USB 3.0 performance enables the R42 to provide images with unparalleled 3-D depth. And the unique EZ-USB flexibility, along with the tough technical back from Cypress, shortened their design cycle."
"Cypress has been a technology leader in USB since 1996, with unparalleled Low-Speed and High-Speed USB solutions. With the addition of the CX3 camera controller to their unique FX3, FX3S and HX3 solutions, they now possess the industry's strongest portfolio of SuperSpeed USB as well," said impress Fu, Senior Director of the USB 3.0 industry Unit at Cypress. "We're excited at the customer response to CX3, as shown by its design win in Raytrix's groundbreaking R42 3-D camera."
Fu added, "The robust reference design kit from e-con Systems extends their design back for CX3. The kit streamlines the design process, making it easier than ever to bring top property HD cameras to market quickly."
"We worked closely with Cypress to develop their new Denebola reference design kit around the CX3 camera controller," said Ashok Babu, President of e-con Systems. "The USB 3.0 camera market is gaining momentum with machine vision, industrial, scientific, medical and consumer products, and CX3 is a tough apt for these applications. With Cypress's circumstantial collateral, designers can exhaust their kit to efficiently create USB Video Class cameras that 'plug and play' with major operating systems."
About the Denebola Reference Design Kit Developed by Cypress partner e-con Systems, the Denebola (See3CAM_CX3RDK) USB 3.0 reference board employs the CX3 controller to interface to an OmniVision OV5640 CMOS image sensor through a 2-lane MIPI CSI-2 interface. It enumerates as a USB Video Class (UVC) compliant camera and works with the UVC device drivers of major operating systems. The RDK includes built-in SPI glint and I2C EEPROM for CX3 firmware storage, as well as a JTAG and RS232 interface for firmware debugging. Designers can exhaust the kit's GPIO headers to connect additional interface boards to add camera control functions such as pan, tilt, zoom, lighting and sync in/out. In addition, designers can furthermore easily supplant the OV5640 CMOS image sensor with any other image sensor by quickly developing just a image sensor daughter board that can breathe plugged on to the Denebola kit and modifying the firmware. E-con furthermore provides additional camera daughter boards as accessories for this Denebola kit. For more information about the Denebola kit or to purchase it, visit www.e-consystems.com/CX3-Reference-Design-Kit.asp.
About EZ-USB CX3 The EZ-USB CX3 programmable USB 3.0 camera controller supports MIPI CSI-2 version 1.01, up to 4 data lanes at hasten up to 1 Gbps per lane, for a total bandwidth of 4 Gbps. CX3 supports many image formats including RAW8/10/12/14, YUV422 (CCIR/ITU 8/10-bit), and RGB888/666/565. Based on the proven EZ-USB FX3™ peripheral controller platform, CX3 comes with an ARM9 CPU and 512 KB SRAM, providing 200 MIPS of computational power for on-chip image data processing. CX3 supports multiple peripheral interfaces such as I2C, SPI, and UART, which can breathe programmed to back Pan, Tilt and Zoom or other camera control functions.
Product Availability The CX3 controller is in full production and is certified by the USB-IF. For more information on EZ-USB SuperSpeed solutions, gratify contact Cypress at usb3@cypress.com.
About Raytrix GmbH German company Raytrix GmbH was founded by Christian Perwass and Lennart Wietzke motivated by the vision to develop and introduce innovative visual media technologies to the industry-, scientific-, and entertainment-market. They are the developer and manufacturer of innovative 4D lightfield cameras and image processing software with incredibly high resolution. Furthermore they develop new computational photography and software products with a straightforward contact to their clients. Learn more at www.raytrix.de.
About Cypress Cypress delivers high-performance, mixed-signal, programmable solutions that provide customers with rapid time-to-market and exceptional system value. Cypress offerings involve the flagship PSoC® 1, PSoC 3, PSoC 4, and PSoC 5LP programmable system-on-chip families. Cypress is the world leader in capacitive user interface solutions including CapSense® palpate sensing, TrueTouch® touchscreens, and trackpad solutions for notebook PCs and peripherals. Cypress is a world leader in USB controllers, which enhance connectivity and performance in a wide scope of consumer and industrial products. Cypress is furthermore the world leader in SRAM and nonvolatile RAM memories. Cypress serves numerous major markets, including consumer, mobile handsets, computation, data communications, automotive, industrial, and military. Cypress trades on the NASDAQ Global Select Market under the ticker symbol CY. Visit Cypress online at www.cypress.com.
Cypress, the Cypress logo, EZ-USB, TrueTouch, PSoC and CapSense are registered trademarks and CX3 and FX3 are trademarks of Cypress Semiconductor Corp. whole other trademarks are the property of their respective owners.


text:
Q: Cant connect from python client to meteor server using python-ddp I am trying to connect meteor server using python-ddp package
https://github.com/hharnisc/python-ddp
When i run the py file,just nothing happens and programme terminates.Server is running port and everything is correct.
   from DDPClient import DDPClient

client = DDPClient('ws://127.0.0.1:3000/websocket')
client.connect()
def connected():
    print("connected")

client.on('connected', connected)

I guess python-ddp package is outdated and thats why its not working ? Is there anyone who is using this package to communicate with meteor server  ?
Do you guys recommend any other package/way to communicate with meteor using python client ?
thanks

redpajama_set_name: RedPajamaStackExchange


Abstract: The goal of this work is studying of the possibilities and properties of the data transfer via power lines. The work deals with remote control of the electrical appliances and problems of data transfer via power line 230 V. The designed devices which provide relative communication via power line are used within the frame of the one family house. Designed devices - concretely input-output unit, PC interface and GSM gate work on a bus principle. The bus is constructed by power line 230 V. Designed devices are composed of microcontrollers AVR and modem TDA5051AT provides their relative communication via power line. Communications protocol with which devices work was developed so that corresponds with hardware resolution of particular devices. Particular devices were practically realized and their activity was checked.
ADÁMEK, Milan, Pavel MARTINEC a Petr NEUMANN. Utilization of power line for remote control of electrical appliances. In: International Conference on Systems - Proceedings [online]. Corfu Island: World Scientific and Engineering Academy and Society (WSEAS), 2010, s. 427-430. [cit. 2019-04-25]. ISSN 1792-4235. Dostupné z: http://www.wseas.us/e-library/conferences/2010/Corfu/SYSTEMS/SYSTEMS2-13.pdf.


text:
Trapping cDNAs encoding secreted proteins from the salivary glands of the malaria vector Anopheles gambiae
Bruno Arcà, Fabrizio Lombardo, Margareth de Lara Capurro, Alessandra della Torre, George Dimopoulos, Anthony A. James, and Mario Coluzzi
PNAS February 16, 1999 96 (4) 1516-1521; https://doi.org/10.1073/pnas.96.4.1516
Bruno Arcà
Fabrizio Lombardo
Margareth de Lara Capurro
Alessandra della Torre
George Dimopoulos
Anthony A. James
Mario Coluzzi
Communicated by Fotis C. Kafatos, European Molecular Biology Laboratory, Heidelberg, Germany (received for review August 7, 1998)
Figures & SI
The signal sequence trap method was used to isolate cDNAs corresponding to proteins containing secretory leader peptides and whose genes are expressed specifically in the salivary glands of the malaria vector Anopheles gambiae. Fifteen unique cDNA fragments, ranging in size from 150 to 550 bp, were isolated and sequenced in a first round of immunoscreening in COS-7 cells. All but one of the cDNAs contained putative signal sequences at their 5′ ends, suggesting that they were likely to encode secreted or transmembrane proteins. Expression analysis by reverse transcription–PCR showed that at least six cDNA fragments were expressed specifically in the salivary glands. Fragments showing a high degree of similarity to D7 and apyrase, two salivary gland-specific genes previously found in Aedes aegypti, were identified. Of interest, three different D7-related cDNAs that are likely to represent a new gene family were found in An. gambiae. Moreover, three salivary gland-specific cDNA fragments that do not show similarity to known proteins in the databases were identified, and the corresponding full length cDNAs were cloned and sequenced. RNA in situ hybridization to whole female salivary glands showed patterns of expression that overlap only in part those observed in the culicine mosquito A. aegypti.
One century after the discovery of the role of mosquitoes in the transmission of Plasmodium parasites, malaria still kills >1.5 million people every year, mostly in sub-Saharan Africa. Malaria control strategies are facing increasing complications because of the acquired resistance of Plasmodium parasites to antimalarial drugs and the growing insecticide-resistance in vector populations. In the last years, research efforts have been focusing on the development of tools for the genetic alteration of mosquito vectors, with the final goal to block the parasite life cycle within mosquitoes, making them incapable of transmitting malaria (1, 2). Recent reports of the successful transformation of the yellow fever mosquito Aedes aegypti (3, 4) make more imminent the prospect of genetically engineering mosquitoes that are refractory to the growth and development of the malaria parasite.
The development of transgenic mosquitoes refractory to malaria transmission requires not only the development of appropriate germ line transformation techniques but also the identification of potentially vulnerable steps in the life cycle of Plasmodium in mosquito and the isolation of promoters able to drive the tissue-specific expression of a chosen gene. Mosquito infection by malaria-causing parasites initiates with the ingestion of gametocytes during blood-feeding on an infected host. Fertilization takes place in the insect midgut, and the mobile zygotes, ookinetes, cross the peritrophic matrix, invade the midgut epithelium, and differentiate into oocysts. Sporozoites develop within the oocysts and then are released into the haemolymph. Haemocoel sporozoites selectively invade salivary glands, where they may be transmitted to a vertebrate host on the next blood feeding (5, 6). The strategy is to produce a transgenic mosquito in which parasite development is blocked at one, or better, two different stages: for example, at the midgut level, interfering with ookinete invasion or oocyst differentiation, and at the salivary gland level, blocking invasion of gland cells or the transmission of infective sporozoites from the glands.
We have focused our initial efforts on the salivary glands of Anopheles gambiae, the most efficient malaria vector in sub-Saharan Africa. The mechanisms by which haemocoel sporozoites locate, recognize, and penetrate salivary glands are presently unknown. However, several observations suggest that specific interactions between ligand(s) on the parasite surface and receptor(s) on the basal lamina and/or plasma membrane of gland cells may be involved in the invasion process. Specific recognition is suggested by the refractoriness to sporozoite invasion of salivary glands exhibited by some mosquito species, presumably because they lack the appropriate receptor (7). Also, the invasion-blocking activity exhibited by antisalivary gland antibodies and by some lectins suggests that sporozoites may interact with glycosylated salivary gland receptor molecules (8). The isolation and characterization of the sporozoite receptor(s) on the mosquito salivary glands may lead to a way to block these interactions: for example, mislocalizing the receptor by expressing its binding domain as a haemolymph-secreted peptide (1).
Salivary glands of mosquito vectors are also important because they express genes whose products are involved in the ability of mosquitoes to feed efficiently on blood. Several secreted factors able to counteract the host haemostatic response, such as vasodilators, inhibitors of platelet aggregation, and anticoagulants, have been identified by classical biochemical techniques in the salivary glands of mosquitoes and other haematophagous insects (9, 10). Six salivary gland-specific genes, four of which are expressed specifically in the female glands, have been isolated and characterized from the mosquito Ae. aegypti (refs. 11 and 12; B. T. Beerntsen, D. E. Champagne, J. Coleman, Y. A. Campos, and A.A.J., unpublished work).
We used a cloning procedure suited to the isolation of sequences that encode secreted and membrane-bound proteins to screen a 5′ end-enriched cDNA library from the salivary glands of the mosquito An. gambiae, and we report here on the identification of fifteen cDNA fragments. All of them, with one exception, appear to have a signal peptide fused to the reporter gene and are likely to encode secreted or membrane-linked proteins. At least six of these cDNA fragments are expressed specifically in the glands, and four of them appear to be female-specific because the corresponding cDNAs cannot be detected in males. These represent the only salivary gland-specific genes identified so far in An. gambiae.
Mosquito Colony and Tissue Dissections.
The An. gambiae strain used for dissections and polytene chromosome preparations was the homokaryotypic GASUA reference strain (Xag, 2R, 2La, 3R, 3L) selected in our insectary in 1989 from a 2Rd-2La polymorphic colony that, in turn, originated from adult females collected in the village of Suakoka Liberia in 1986. Salivary glands from 0- to 3-day-old An. gambiae adult females were dissected in PBS and were kept on ice for a maximum of 1 hour. After a low speed centrifugation step, the medium was removed, and the glands were frozen in liquid nitrogen and were stored at −80°C. Anterior portions of the thorax containing the salivary glands were dissected from 4-day-old females by a surgical blade, were placed in an Eppendorf tube on dry ice, and were stored at −80°C.
RNA Purification.
General nucleic acids manipulations were performed according to standard procedures (13, 14) if not otherwise specified. Poly(A)+ RNA from the salivary glands of 0- to 3-day-old females and from thoracic tissues of 4-day-old females was purified by, respectively, Dynabeads oligo(dT)25 (Dynal, Oslo) and the Oligotex Direct mRNA Kit (Qiagen, Chatsworth, CA).
cDNA Library Construction.
The thoracic cDNA library was constructed by using 7 μg of poly(A)+ RNA and the ZAP Express kit (Stratagene). A PCR-based method was used for the screening of the thoracic cDNA library (15). The signal sequence-enriched cDNA library was constructed as described (16) with minor modifications. First-strand cDNA was synthesized by using 5 ng of random hexamers and ≈1 μg of salivary gland poly(A)+ RNA. Reaction conditions for the addition of 10–20 deoxycytosine residues to the 3′ end of the first strand cDNA were determined empirically by using a 5′ end-labeled 25-mer oligonucleotide. Second strand cDNA was synthesized by the ESLG primer containing an EcoRI linker and an oligo(dG) (5′-GCGGCCGCGAATTCTGACTAACTGAC(G)17-3′). After second strand synthesis, the SacI adapters were ligated, and fragments in the range of 200 to 600 bp were size-selected by gel electrophoresis. These fragments were amplified (25 cycles of 94°C, 30 sec; 49°C, 1 min; and 72°C, 3 min) by using the primers LLHES (5′-GAGGTACAAGCTTGATATCGAGCTCGCGG-3′) and ESL (5′-GCCGCGAATTCTGACTAACTGAC-3′). Amplified cDNA fragments were purified by gel electrophoresis, were digested with SacI and EcoRI, and were cloned directionally into the EcoRI- and SacI-digested pcDL-SRα-Tac(3′) vector (kindly provided by T. Honjo, Kyoto University). An aliquot of the cDNA library was used to transform XL1-Blue super-competent cells (Stratagene).
Screening and Immunostaining.
Forty-nine individual colonies were transferred from the master plate to an agar plate in a matrix format of seven rows by seven columns and were designated as a pool. Five-hundred nanograms of plasmid DNA preparation from each pool were used for transfection of 105 COS-7 cells by the LipofectAmine Reagent (GIBCO/BRL). Forty-eight to seventy-two hours after transfection, the cells were stained with the fluorescein isothiocyanate-conjugated anti-human interleukin 2 receptor (Anti-Tac) mAb (Dako), and the immunofluorescent positive pools were identified microscopically. Positive pools were divided in 14 smaller pools and were rescreened, leading to the identification of single positive clones. Pools were judged as positive or negative by comparison to transfections with the positive and negative control plasmids pcDL-SRα-hGCSF(5′)-Tac(3′) and pcDL-SRα-hRAR(5′)-Tac(3′) (16).
Signal Peptide Prediction and Sequence Comparison.
cDNA clones were sequenced by the T3, T7, SRA (5′-TTTACTTCTAGGCCTGACG-3′), TAC (5′-CCATGGCTTTGAATGTGGCG-3′), and SLG9 (5′-GACTAACTGACGGGGGGGGG-3′) primers. Signal peptide prediction analysis was performed by using the psort worldwide web server (http://psort.nibb.ac.jp/). Sequence comparison was performed by using the wisconsin package 9.1 (Genetics Computer Group, Madison, WI) and the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov/) and the European Bioinformatics Institute (http://www.ebi.ac.uk/ebi_home.html) worldwide web servers.
In Situ Hybridization to Polytene Chromosomes.
Polytene chromosome spreads were obtained from half-gravid female mosquitoes (17), and probe-labeling and in situ hybridization procedures were performed as described (18). Sites of hybridization were localized to specific bands within numbered and lettered subdivisions of the polytene chromosome map (M.C., A. Sabatini, M. A. Di Deco, and V. Petrarca, unpublished work) revised from that of Coluzzi and Sabatini (19).
Expression Analysis by Reverse Transcription (RT)–PCR.
About 100 ng of DNase-treated total RNA (DNAaseI RNase-free, Boehringer Mannheim) was used for the RT-PCR amplifications with gene-specific primers and the SuperScript one-step RT-PCR system (GIBCO/BRL). After the reverse transcription step (50°C for 30 min) and heat-inactivation of the reverse transcriptase (2 min at 94°C), 35 cycles were performed as follows: 94°C, 30 sec; 55°C, 30 sec; and 72°C, 45 sec. The sequence of the gene-specific oligonucleotide primers is not reported because of space limitations and will be made available on request.
In Situ Hybridization to Salivary Glands.
The cDNA clones of interest were subcloned as SacI-EcoRI fragments into the polylinker of the plasmid vector pBCKS(+) (Stratagene). Antisense and sense RNA probes were synthesized by using digoxigenin-substituted ribonucleotides. Salivary glands were dissected from 3- to 7-day-old adult female mosquitoes and were fixed in 4% paraformaldehyde for 30 min in PBS supplemented with 67 mM EGTA. The glands were dehydrated in four washes of methanol followed by five washes in 100% ethanol and were stored at −20°C in ethanol for several days before use. Glands were postfixed for 30 min in 5% formaldehyde in PBS with 0.1% Tween 20, were digested for 10 min in 33 μg/ml proteinase K, were rinsed twice in PBS with 0.1% Tween 20 to stop proteinase digestion, and, finally, again were postfixed for 30 min in 5% formaldehyde. Hybridizations were carried out in the presence of formamide, and an alkaline wash with 0.1 M Tris (pH 9.5) was used to remove background. Preparations were visualized and photographed with compound microscopy.
Signal Sequence Trap (SST).
To identify cDNAs encoding secreted and membrane-bound proteins specifically expressed in the salivary glands of the malaria vector An. gambiae, we used the SST method. The SST technique originally was developed by Tashiro et al. (16) to take advantage of the observation that secreted and transmembrane proteins are targeted to the secretory pathway by amino-terminal signal peptides that are encoded by signal sequences, usually located within 400 base pairs of the 5′ termini of the mRNAs. In the SST system, a 5′ end-enriched cDNA expression library is cloned directionally between a promoter that confers high level of expression in COS-7 cells and a "reporter" gene, the α chain of the human interleukin 2 receptor (Tac). cDNAs that are cloned in-frame with Tac and contain signal sequence produce fusion proteins that can be expressed on the surface of transfected COS-7 cells and are easily detectable by immunostaining with an anti-Tac monoclonal antibody.
Salivary glands dissected from An. gambiae females were used to construct a 5′ end-enriched cDNA expression library. Fragments from 200 to 600 bp in length were cloned directionally in the pcDL-SRα-Tac(3′) vector (16). Twenty-two pools (each one consisting of 49 clones) were screened initially by transfection in COS-7 cells, and 21 of them were positive after immunostaining with the anti-Tac mAb. Six pools were chosen randomly, were divided into subpools, and were rescreened in COS-7 cells, leading to the isolation of 18 immunofluorescent positive clones. Sequence analysis showed that 15 cDNA fragments were unique and that their size ranged from 164 to 523 bp in length (Table 1). Three additional cDNA fragments represented shorter versions of the clones aB1, cE5, and iC5 and were not characterized further. The putative peptide sequences were deduced from the ORFs fused to Tac, assuming that the first methionine was the start codon. Scores obtained by the von Heijne (20) and McGeoch (21) methods are reported in Table 1. At least 12 of the 15 cDNA fragments have large positive scores with both methods, suggesting that they likely encode secreted or membrane-bound proteins.
Properties of the cDNAs isolated by SST
Only half of the clones gave significant similarity to previously identified proteins when the sequences of the cloned cDNA fragments were used for database searches (Table 1). Five different cDNAs showed similarity to D7 and apyrase, two genes that are specifically expressed in the female salivary glands of the yellow fever mosquito Ae. aegypti (22–24). The cDNA fragment dG5 encoded an opsin; the cF6 clone shared similarity with venom allergens from ants and wasps (25), and we will refer to it as gVAG.
Anopheles gambiae Apyrase and 5′-Nucleotidase.
The clones cF3 and iC6 both showed similarity to the Ae. aegypti apyrase and to several members of the 5′-nucleotidase family. An amino acid alignment of the An. gambiae cF3 and iC6 cDNA fragments with the Ae. aegypti apyrase is shown in Fig. 1A. The two clones show, respectively, a 45 and 56% identity and a 70 and 67% similarity to the Ae. aegypti gene products. The clones also share high similarity, 50 and 65%, respectively, with members of the 5′-nucleotidase family (data not shown).
Alignment of the Ae. aegypti apyrase (A) and D7 (B) proteins to the translated An. gambiae cDNA fragments identified by the SST. Asterisks mark identities in all of the sequences. Dots identify conservative substitutions. Identities in at least two of the aligned sequences are shaded. In B, the putative signal peptides of the D7-related cDNA fragments are boxed with a thick line.
D7-Related.
The cDNA fragments dB1, iB6, and iC5 show similarity to D7, a secreted protein that is expressed abundantly and exclusively in the salivary glands of Ae. aegypti adult females (22). The D7 function is unknown, but the stage-, sex-, and tissue-specific patterns of expression suggest that it may have a role in blood feeding. Because of the sequence similarity to the Ae. aegypti gene, we propose to refer to these cDNAs as D7-related and to identify them individually as D7r1 (dB1), D7r2 (iB6), and D7r3 (iC5). Fig. 1B shows an alignment of the deduced peptide sequence of these three partial cDNAs with the Ae. aegypti D7. D7r1, with a 35% identity and a 52% similarity, is the most similar to D7 whereas D7r2 and D7r3 show, respectively, a 23 and 26% identity (44 and 45% similarity) to the Ae. aegypti protein. D7r2 and D7r3 share 62% identical amino acid residues between themselves and are by far the most closely related to each other. These three An. gambiae cDNA clones hybridize relatively closely to chromosomal positions on the right arm of the third chromosome in the divisions 30A (D7r2) and 30B (D7r1 and D7r3) (Table 1).
cDNA Clones Expressed Specifically in the Salivary Glands.
To evaluate the tissue-specific expression of the 15 cDNAs identified by SST, we amplified the corresponding messenger RNAs by RT-PCR using, as template, total RNA extracted from female salivary glands and carcasses (adult females with salivary glands removed) and from adult males. As summarized in Fig. 2, at least six cDNAs were specifically expressed in the salivary glands, and five other clones may be expressed at higher level in female glands than in other tissues or in males.
RT-PCR analysis of the 15 An. gambiae cDNA fragments. sg, salivary glands of adult females; c, carcasses (adult females with salivary glands removed); m, adult males. The different cDNA clones are indicated on the right side. rpS7, ribosomal protein S7. The specificity of expression is annotated as follows: ●, specifically in female salivary glands; ○, in female salivary glands and males; +, enriched in female salivary glands.
RT-PCR analysis clearly showed that cF3 is specifically expressed in female glands whereas iC6 is detectable also in carcasses and in adult males (Fig. 2), suggesting that cF3 is most likely the An. gambiae apyrase whereas iC6 may represent a 5′-nucleotidase. The three D7-related cDNA fragments all showed high levels of expression in female salivary glands. However, although D7r1 and D7r2 are specifically expressed in female glands, D7r3 is detectable also at a very low level when RNA from carcasses is used as template (Fig. 2).
Finally, three cDNAs, aA5, aB1, and cF1, were salivary gland-specific (Fig. 2). The aA5 cDNA was detectable only when female salivary gland RNA was used as template whereas aB1 and cF1 were detectable also in males. The simplest interpretation is that aA5 is expressed only in female glands and aB1 and cF1 are expressed also in male glands. As previously mentioned, these three cDNA fragments did not show significant similarity to any other protein in the databases. We screened a thoracic cDNA library and cloned the full length cDNAs corresponding to aA5, aB1, and cF1. They are, respectively, 1,372, 436, and 721 bp in length and encode putative proteins of 401, 114, and 189 amino acids. We propose to refer to these cDNAs as gSG1 (aA5), gSG2 (aB1), and gSG3 (cF1), for gambiae salivary genes 1, 2, and 3 (Fig. 3). In all three cases, the additional sequence information did not allow further insight into their possible function. gSG2 is a small protein, rich in glycine and phenylalanine, with a highly hydrophobic profile (data not shown). The gSG3 putative protein contains a serine-rich domain toward the amino terminus and, at the carboxy terminus, a threonine-rich region composed of repeats of the motif EEATTT or closely related hexapeptides (Fig. 3). Other, less obvious, repeats are found between the serine- and threonine-rich regions. The functional significance of these motifs is still to be elucidated.
Peptide sequences deduced from the gSG1, gSG2, and gSG3 cDNA clones are shown from the top to the bottom. The putative signal peptides at the amino terminus are underlined. In the gSG3 sequence, the serine-rich region is boxed and shaded, the motif RP(P/W)(F/W) is underlined by a thick line, and the repeated motifs at the carboxy terminus are boxed.
Hybridization in Situ to Salivary Glands.
A selected set of cDNAs, which includes the six salivary gland-specific clones, were expressed as antisense and sense RNA probes and were hybridized in situ to female salivary glands to determine the localization of expression of their corresponding genes (Table 2). In all cases, signals were obtained only with antisense RNA probes and not with control sense RNA probes. The apyrase-like cDNA cF3 hybridizes to mRNA localized in the distal–lateral lobes with the signal extending along the length of the proximal–lateral lobes (Fig. 4). In addition, hybridization was seen with this probe to mRNA in some of the proximal regions of the medial lobe. The three D7-related cDNAs hybridize to mRNA in the distal-lateral lobes, with occasional hybridization to the medial lobe. The gVAG probe also hybridizes to the distal-lateral lobes of the female glands. The cDNA gSG1 is peculiar because it is specifically expressed in female glands and hybridizes to mRNA in subsets of cells in the medial lobes of female glands. Finally, two cDNAs, gSG2 and gSG3, with sequences that are expressed in both males and females, are expressed specifically in the proximal–lateral lobes of female glands. The gSG2 cDNA also hybridizes occasionally to the proximal medial lobe.
Expression patterns of genes in adult female salivary glands
Hybridizations in situ to adult female salivary glands with antisense probes as indicated. C, control glands hybridized with a sense probe; pL, proximal-lateral lobes; dL, distal-lateral lobes; M, medial lobe. The arrow points to the region of the proximal–lateral lobes that hybridizes to the apyrase-like cF3 probe. The gSG2 probe hybridizes to the proximal–lateral lobes (upper gland) and to the proximal–medial lobe (lower gland). Note limited signal over the medial lobe in the hybridization with the gSG1 probe.
We constructed a 5′ end-enriched cDNA library from the salivary glands of the malaria vector An. gambiae and used the SST technique to screen for cDNAs coding for secreted and type I membrane proteins. The SST method worked efficiently with the salivary glands, and 6% of the clones analyzed were positive. Taking into consideration that only one cDNA fragment in every three is expected to be cloned in frame with the reporter gene, the average number of cDNAs containing a signal peptide is estimated to be 18%. This number is relatively high but perhaps not surprising for a tissue involved in intense secretory activity. Moreover, the 5′ end-enriched salivary gland cDNA library was shown to be complex, with 15 of the 18 clones isolated being unique. In addition, insect signal peptides were efficiently recognized in COS-7 cells, as shown both by the signal peptide prediction methods and by the sequence comparison analysis. At least 6 of the 15 cDNA clones identified were expressed specifically in the salivary glands.
Two cDNA fragments showed similarity to the Ae. aegypti apyrase and to several members of the 5′-nucleotidase family (24). Apyrase activity is generally rare in animal tissues, and salivary glands of haematophagous insects are the richest source known. The apyrase is a secreted protein that hydrolizes ATP and ADP to AMP and Pi. Its function is to help mosquitoes blood-feed efficiently by inhibiting the ADP-induced platelet recruitment and aggregation. 5′-nucleotidases are membrane-linked ubiquitous proteins that hydrolyze extra cellular nucleotides to nucleosides making them permeable to cell membranes. It has been suggested that the apyrase, a secreted protein, evolved by gene duplication and subsequent divergent evolution from the membrane-bound 5′-nucleotidase by loss of a carboxy terminal domain involved in membrane anchoring (24). We were able to show by RT-PCR that one of these cDNAs, cF3, is expressed only in the female salivary glands of An. gambiae whereas the other one, iC6, is also detectable, at a lower level, when RNA from carcasses or from adult males was used as template. These patterns of expression strongly suggest that cF3 may be the An. gambiae apyrase and that iC6 may be a 5′-nucleotidase. The identification of cDNA fragments corresponding to these genes in An. gambiae offers the opportunity to investigate the evolutionary relationship between these two genes within the same organism.
We also have identified three members of a family of genes related to the Ae. aegypti D7. Two cDNAs, D7r1 and D7r2, were expressed specifically in female salivary glands whereas the third, D7r3, also was detectable at a very low level when RNA from carcasses was used as template. It is possible that D7r3 is expressed in female glands and, at a much lower level, in some other female tissue. However, because of the apparently higher abundance of the corresponding mRNA as compared with D7r1 and D7r2 (Fig. 2), we cannot rule out the possibility that the weak band detectable from carcass RNA preparations is attributable to a small contamination with gland tissue. In any case, the redundancy in copy number of these D7-like genes suggests that they must play some essential role in An. gambiae, most probably in connection with blood feeding.
The Ae. aegypti D7 gene is encoded in five exons separated by small introns with the signal peptide that is encoded by exon 1 (22). As shown in Fig. 1B, the An. gambiae D7-related cDNA fragments can be aligned for their entire length to the carboxy terminal half of the D7 protein with the similarity starting immediately after the signal peptides and involving a region corresponding to exons 4 and 5. If their function is homologous to the one that D7 plays in Ae. aegypti, then the domain(s) encoded by exons 4 and 5 of D7 must be especially important. The cloning of the full length cDNAs and their molecular characterization may help elucidate the function and the structure of this intriguing protein family.
The efficacy of the SST technique as applied to the salivary glands of An. gambiae is confirmed further by the identification of the three salivary gland-specific cDNAs gSG1, gSG2, and gSG3. Two of them are expressed both in male and female glands, but their sequence is unrelated to Maltase-like I (26) and Amylase I (27), the only genes previously identified as expressed in both male and female Ae. aegypti salivary glands.
The cDNA clones bB2, cB1, gVAG, and dF2 showed a pattern of expression that may be compatible with higher expression in female glands than in other tissues or in males. The gVAG cDNA fragment showed similarity to allergens from ants and wasps (25). These allergens belong to a family of secreted proteins whose functions are still poorly understood and is sometimes named the CAP family (28) because it counts members from mammals (cysteine-rich secretory proteins), insects (AG5/AG3), and plants (pathogenesis-related proteins). Finally, the dG5 cDNA fragment, which shows very high similarity to opsins, most likely represents a cloning artifact caused by tissue contamination that occurred during dissection because opsin genes are characteristically expressed in the eye.
Hybridization of cDNA probes to whole female salivary glands produced a number of interesting results. In general, the pattern of expression was consistent with what is seen in the culicine mosquito, Ae. aegypti (11). The cDNAs corresponding to genes expressed both in males and females, gSG2 and gSG3, are expressed in the proximal–lateral lobes. This expression pattern reflects that of two genes, Maltase-like I (26) and Amylase I (27), that are expressed in both male and female Ae. aegypti and whose products most likely are involved in sugar feeding. The proximal–lateral lobes of female salivary glands are postulated to overlap functionally the complete male salivary glands (11). Expression of the gene corresponding to gSG2 in the proximal–medial lobe was not anticipated based on its sex-specific expression and may reflect some fundamental differences in the organization of the salivary glands between anopheline and culicine mosquitoes.
Genes expressed specifically in the female salivary glands of Ae. aegypti are expressed only in the distal–lateral and medial lobes (11). Consistent with this observation, the three cDNAs D7r1, D7r2, and D7r3, related by sequence structure to the D7 gene of Ae. aegypti (22), are expressed in the distal–lateral lobes of the female salivary glands. However, we could detect clear signal corresponding to expression in the medial lobe with the D7r1 cDNA only. Similarly, the cDNA cF3, with a conceptual translation product similar to the apyrase gene of Ae. aegypti (23, 24), also shows expression in the distal-lateral lobes and not in the medial lobe. In addition, it appears to be expressed in some regions of the proximal–lateral lobes. This pattern is unexpected from the results of Ae. aegypti, but, considering the long evolutionary separation of the families to which these species belong, perhaps it is not so unlikely that the biology and gene expression in the glands is different.
The expression pattern of the gSG1 cDNA is remarkable in that it is expressed only in the medial lobe of female salivary glands. The sialokinin gene of Ae. aegypti was shown by hybridization in situ to be expressed only in the medial lobe (B. T. Beerntsen, D. E. Champagne, J. Coleman, Y. A. Campos, and A.A.J., unpublished work). However, there is no similarity between the conceptual translation product of gSG1 and the sialokinin gene. Furthermore, we were never able to observe hybridization of the gSG1 probe to the whole medial lobe. Although it is possible that gene expression in the medial lobe cannot be fully evaluated because it is refractory to our hybridization technology, it is possible that only variable but distinct regions of the medial lobe actually express the corresponding gene.
In conclusion, we applied the SST method to the salivary glands of An. gambiae, and we were able to identify at least six genes specifically expressed in this organ. These represent the first salivary gland-specific cDNAs identified from the most efficient malaria vector in sub-Saharan Africa. The isolation of the corresponding genes should provide tissue-specific promoters that may be helpful for the development of future vector control strategies. The efficiency of the trapping method and the low number of clones that were screened suggest that a more exhaustive screening of the library should allow for the identification of several other tissue-specific genes. This will certainly result in a better knowledge of the salivary gland functions and will allow a deeper understanding of the molecular interactions occurring both between Plasmodium and mosquito and between mosquito and the vertebrate host.
We thank Prof. Christos Louis (Institute of Molecular Biology and Biotechnology, Crete, Greece) for the suggestions, the discussions, and the encouragement since the early stages of this work; Prof. F. C. Kafatos (European Molecular Biology Laboratory, Heidelberg) for helpful comments on an earlier version of the manuscript; Prof. Tasuku Honjo (Kyoto University) for kindly providing the pcDL-SRα-hRAR(5′)-Tac(3′) and pcDL-SRα-hGCSF(5′)-Tac(3′) plasmid vectors and the detailed protocols for the SST technique; Dr. Manlio Di Cristina for the help with cell culture; Gianni Pietrangeli and Maria Calzetta for the expert technical assistance; and L. Olson for helping in typing the manuscript. B.A. and A.d.T. were supported by postdoctoral fellowships of the Fondazione "Istituto Pasteur-Cenci Bolognetti". G.D. was supported by a Training and Mobility of Researchers postdoctoral fellowship. This work was supported by the European Union Grant ERBFMRXCT960017 to M.C. and grants from the Burroughs-Wellcome Fund and John D. and Catherine T. MacArthur Foundation to A.A.J.
↵† To whom reprint requests should be addressed at: Istituto di Parassitologia, Universitá di Roma "La Sapienza," Piazzale Aldo Moro 5, 00185 Rome, Italy. e-mail: b.arca{at}caspur.it.
Data deposition: The sequences reported in this paper have been deposited in the European Molecular Biology Laboratory database (accession nos. AJ000034–AJ000037, Y17688–Y17690, Y17699–Y17705, Y17717, and AJ130949–AJ130951).
reverse transcription;
SST,
signal sequence trap
Received August 7, 1998.
Accepted November 30, 1998.
Copyright © 1999, The National Academy of Sciences
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You are going to email the following Trapping cDNAs encoding secreted proteins from the salivary glands of the malaria vector Anopheles gambiae
Bruno Arcà, Fabrizio Lombardo, Margareth de Lara Capurro, Alessandra della Torre, George Dimopoulos, Anthony A. James, Mario Coluzzi
Proceedings of the National Academy of Sciences Feb 1999, 96 (4) 1516-1521; DOI: 10.1073/pnas.96.4.1516

redpajama_set_name: RedPajamaCommonCrawl


text:
Pulis rules out making a snap decision over his Middlesbrough future
Pulis waiting on his future
By Scott Wilson scottwilsonecho Chief Sports Writer
SHEFFIELD, UK 13TH FEBRUARY Tony Pulis the Middlesbrough manager during the Sky Bet Championship match between Sheffield United and Middlesbrough at Bramall Lane, Sheffield on Wednesday 13th February 2019. (Credit: Mark Fletcher | MI News)
TONY PULIS will not be making a snap decision over his future no matters what happens to Middlesbrough's play-off hopes in the final game of the season tomorrow afternoon.
Boro head to Rotherham tomorrow lunch-time knowing a victory could see them leapfrog Derby to claim the final play-off spot if the Rams fail to win their final fixture at home to West Brom.
The Teessiders would scrape into the play-offs with a draw if Derby were to lose by seven or more goals and Bristol City were to fail to beat Hull City, but the reality is that Boro have to win at the New York Stadium to have any chance of finishing in the top six. If Derby win, however, their result will be irrelevant.
If Boro were to fail to make the play-offs, attention would immediately switch to Pulis' future as manager. The 61-year-old's contract is due to expire this summer, and while there is an automatic one-year extension clause that would be triggered if all parties agreed, he has consistently refused to discuss his plans beyond the end of the current campaign.
That stance did not shift yesterday, with Pulis insisting he is yet to decide on his future. Whatever happens tomorrow, the Boro boss intends to sit down with Steve Gibson once the dust has settled to discuss his plans.
"Whatever happens on Sunday, I won't be coming in and making a decision on my future," said Pulis. "I'll wait to have a conversation with Steve. I'm hoping he'll take me out for something to eat, so we'll see what happens from there.
"I haven't made my mind up until I speak to Steve. If people want to take that a certain way, that's up to them. I'll be talking to Steve. I have tremendous respect for the chairman, as a chairman and a as a person. So I won't be discussing it with anyone else – not even my wife at the moment.
"The supporters should realise how fortunate and lucky they are to have someone like Steve. My respect has always been for the person who has been doing all this, and that's Steve Gibson. I'll speak to Steve before I make any decision."
However, whether he remains in charge or not, Pulis insists there can be no return of the 'boom and bust' mentality that saw Boro squander a fortune in the immediate aftermath of their relegation from the Premier League.
"We've got to decide which way we're going to go," he said. "There's been a lot of money spent here, and it needed to change. Some people don't see it, but you've only got to open your eyes to see what we've done this year and how we've done things."
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