250-323 exam Dumps Source : Data Protection Administration for(R) Windows using NBU 5.0
Test Code : 250-323
Test name : Data Protection Administration for(R) Windows using NBU 5.0
Vendor name : Symantec
braindumps : 201 real Questions
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MOUNTAIN VIEW, Calif.--(enterprise WIRE)--Symantec Corp. (NASDAQ: SYMC), the area’s leading cyber security business, these days introduced that greater than 120 agencies absorb joined forces with Symantec to coerce down the can impregnate and complexity of cyber safety, whereas improving response times to protect companies against subtle threats. This includes primary avid gamers fancy AWS, box, IBM safety, Microsoft, Oracle, ServiceNow and Splunk, in addition to dozens of different technology innovators, who are now edifice or offering more than 250 products and capabilities that combine with Symantec’s built-in Cyber defense (ICD) Platform.
“The probability landscape every of us pan isn't static and is continually on the flow, as are their customers and personnel, so techniques to deal with that variety of ambiance are very complicated”Tweet this
This remarkable business collaboration reflects a “platform shift” in the cyber security trade, as current analysis from enterprise approach community (ESG) indicates enterprise consumers want to consolidate providers and undertake more integrated structures backed with the aid of an open ecosystem.
built-in defense improves security by means of expanding the velocity and effectiveness, whereas drastically decreasing the supplies required. To create that shift even simpler, Symantec moreover nowadays introduced vital innovations – together with a current widely wide-spread statistics exchange, shared administration capabilities, and upgraded information loss prevention utility that advocate purchasers stop untrusted apps before they compromise exclusive information. every are built on Symantec’s ICD Platform, which provides a unified framework for assistance protection, risk insurance policy, identity administration and compliance across endpoints, networks, purposes, and clouds.
“There’s a seismic shift going on in cyber protection,” observed artwork Gilliland, EVP and GM business items, Symantec. “The ancient means of combating cyber-attacks the employ of fragmented tools has revolve into too complicated and costly to control. built-in structures are the long run. We’re arrogant to breathe main this platform shift with a transparent vision and successful portfolio – along with a all bunch of companions and thousands of experts working each day on the entrance traces to give protection to their customers. we're completely convinced that their finest protection going ahead is an integrated protection.”
New analysis Demonstrates the exigency for integrated structures
ESG currently published current client research showcasing how the inability of a cohesive safety technology strategy creates real issues for companies, leading clients to are seeking greater integrated systems and fewer, extra strategic vendors. Key findings in line with the analysis consist of:
“practically two-thirds of great firms surveyed employ as a minimum 25 distinctive cyber security products. For safety operations facilities, managing disparate materiel can breathe ineffective, costly, and time drinking, above every considering the fact that the shortage of cyber security talents,” talked about Jon Oltsik, senior predominant analyst and fellow, ESG. “This explains why CISOs wish to consolidate and integrate their security infrastructure with systems and open architectures that provide superior developer advocate and convey a accomplice ecosystem with potent third-birthday party integrations.”
Symantec Integrates items, functions and companions
Symantec began constructing ICD two and a half years in the past with its acquisition of Blue Coat systems, which introduced most effective-of-breed net and cloud safety applied sciences to Symantec’s market-main endpoint, email and data loss prevention (DLP) technologies. at the time, Symantec saw and heard that purchasers absorb been spending gigantic time and supplies to integrate factor applied sciences with a purpose to derive real cost from their cyber security investments. So, the enterprise invested in a methodology and roadmap to bring an integrated platform that greatly reduces can impregnate of operations whereas enhancing the velocity and accuracy of prevention, detection and response in an application to gash back risk.
since then, Symantec has:
ICD Platform Earns vast Ecosystem guide
Symantec now has greater than a hundred and twenty partners in TIPP, who're edifice or providing greater than 250 current purposes and features that combine with Symantec’s ICD Platform, so commercial enterprise customers can reduce the impregnate and complexity of their protection operations. in addition, Symantec is now launching a current “Innovation Playground” software within TIPP to simplify integration with startups. the brand current program will enable startup groups to leverage Symantec APIs and gain access to items, engineering substances, and customer innovation days.
“as a route to gash back safety operations complexity and combat today’s more and more refined adversary, groups want products that labor as a platform as an alternative of a military of point items working in silos,” referred to Oliver Friedrichs, VP of safety automation and orchestration at Splunk. “Splunk’s aid for the ICD Platform gives their joint clients with consolidated views throughout their protection infrastructure, including incidents flowing from endpoint, web, network and electronic mail security options.”
“At box, safety is a precise priority and they are committed to providing their purchasers with effective controls to give protection to their sensitive content material,” talked about Niall Wall, senior vice chairman of partners at field. “Symantec is a founding member of the domain reliance Ecosystem. we're excited about Symantec’s built-in Cyber defense and the route it's going to advocate our mutual purchasers in the reduction of risk of statistics loss, notice advanced threats, and seamlessly collect their protection capabilities.”
“safety analysts these days cope with increasingly complicated threats, fragmented safety equipment, and siloed businesses,” said wealthy Telljohann, director of enterprise edifice at IBM safety. “To combat this they are since the cyber protection landscape is disturbing a shift to integrated platforms with a view to reduce complexity and value. we now absorb constructed an integration using Symantec ICD change, so the IBM Resilient Incident Response Platform can provide shrewd orchestration, automation, and enrichment of incidents prompted with the aid of Symantec ICD, allowing analysts to respond intelligently to threats.”
ICD Platform Drives customer Adoption and cost
As evidenced by Symantec’s fresh quarterly salary, further and further consumers are relocating past individual items to undertake the ICD platform and portfolio. as an instance:
“The hazard panorama they every pan is not static and is at every times on the flow, as are their consumers and employees, so ideas to cope with that sort of environment are very complicated,” spoke of Emily Heath, vice president and chief tips safety officer, United airlines. “Visibility of your atmosphere and integration of solutions are a key a portion of that method. as an example, if one security wield catches something, it is an Awful lot greater productive for us if those controls are built-in and may talk seamlessly with each other to befriend with real time detection. additionally, if security suppliers retract the time to integrate throughout the stack so they don’t should, that results in an excellent better influence.”
Symantec Extends ICD Platform with current facets & features
Symantec is introducing three current know-how improvements these days that prolong ICD for shared intelligence and shared administration across multiple technology add-ons, in addition to current “possibility mindful” data coverage capabilities:
For more counsel on ICD, gladden talk over with: www.symantec.com/icd.
For more details about TIPP, gladden hunt recommendation from: https://www.symantec.com/expertise-partners.
For greater details on how Symantec and companions are leading the shift to integrated Cyber protection, gladden consult with: https://www.symantec.com/blogs/function-experiences/united-we-stand
For extra details on the cyber safety platform shift, gladden visit: https://www.symantec.com/blogs/feature-reviews/cyber-security-platform-shift-more-at ease-less-complicated
to peruse a detailed mapping of Symantec product and accomplice integrations, gladden hunt recommendation from: https://www.symantec.com/integration.
Symantec company (NASDAQ: SYMC), the realm's main cyber safety business, helps companies, governments and individuals cozy their most necessary data anywhere it lives. organizations internationally materialize to Symantec for strategic, built-in options to peruse after towards refined attacks across endpoints, cloud and infrastructure. Likewise, a global group of more than 50 million americans and families weigh on Symantec's Norton and LifeLock product suites to protect their digital lives at home and across their instruments. Symantec operates one of the most world's largest civilian cyber intelligence networks, permitting it to note and offer protection to towards probably the most advanced threats. For more information, gladden consult with www.symantec.com or unite with us on facebook, Twitter, and LinkedIn.
observe TO U.S. EDITORS: if you would fancy additional info on Symantec corporation and its products, gladden consult with the Symantec Newsroom at http://www.symantec.com/information.
Symantec and the Symantec logo are trademarks or registered trademarks of Symantec supplier or its associates within the U.S. and different nations. different names can breathe emblems of their respective house owners.
forward-looking STATEMENTS: Any forward-looking indication of plans for products is prefatory and every future unlock dates are tentative and are discipline to exchange. Any future unlock of the product or deliberate adjustments to product skill, performance, or characteristic are domain to ongoing comparison by means of Symantec, and might or may moreover now not breathe applied and may now not breathe regarded company commitments with the aid of Symantec and should now not breathe relied upon in making purchasing decisions.
1 supply: ESG research, hazard Detection and Response Survey, December 2018.
2 source: ESG analysis document, 2019 technology Spending Intentions Survey, February 2019.
three supply: ESG master Survey results, Cybersecurity panorama: The Evolution of business-classification companies and systems, October 2018.
MOUNTAIN VIEW, Calif.--(business WIRE)--
greater than 120 companions, together with AWS, field, IBM safety, Microsoft, Oracle, ServiceNow and Splunk, entrust to Symantec’s built-in Cyber protection Platform
Symantec Corp. (SYMC), the world’s main cyber security enterprise, nowadays announced that more than a hundred and twenty corporations absorb joined forces with Symantec to drive down the impregnate and complexity of cyber protection, while improving response instances to protect organisations in opposition t refined threats. This comprises principal players fancy AWS, container, IBM safety, Microsoft, Oracle, ServiceNow and Splunk, as well as dozens of alternative know-how innovators, who are now constructing or supplying more than 250 items and services that combine with Symantec’s built-in Cyber defense (ICD) Platform.
This remarkable business collaboration displays a “platform shift” within the cyber protection industry, as current research from enterprise strategy group (ESG) suggests business customers want to consolidate providers and adopt greater integrated structures backed via an open ecosystem.
integrated protection improves safety through expanding the hurry and effectiveness, whereas enormously cutting back the supplies required. To create that shift even easier, Symantec moreover nowadays announced essential innovations – including a brand current ordinary data change, shared management capabilities, and upgraded information loss prevention utility that befriend customers cease untrusted apps before they compromise exclusive information. every are developed on Symantec’s ICD Platform, which provides a unified framework for counsel insurance policy, danger coverage, id management and compliance across endpoints, networks, purposes, and clouds.
“There’s a seismic shift occurring in cyber safety,” mentioned technique Gilliland, EVP and GM enterprise products, Symantec. “The historical system of fighting cyber-assaults the employ of fragmented materiel has become too complicated and costly to control. integrated platforms are the longer term. We’re arrogant to breathe main this platform shift with a transparent vision and successful portfolio – together with a all lot of companions and thousands of specialists working daily on the front lines to offer protection to their customers. we're fully convinced that their top-quality protection going ahead is an built-in defense.”
New analysis Demonstrates the want for built-in structures
ESG currently posted current consumer research showcasing how the inability of a cohesive safety technology strategy creates actual problems for agencies, main valued clientele to are looking for greater built-in platforms and fewer, more strategic providers. Key findings in line with the analysis include:
“well-nigh two-thirds of tremendous firms surveyed employ at the least 25 distinctive cyber protection items. For security operations centers, managing disparate materiel can breathe ineffective, expensive, and time drinking, certainly because the scarcity of cyber protection competencies,” pointed out Jon Oltsik, senior necessary analyst and fellow, ESG. “This explains why CISOs wish to consolidate and combine their protection infrastructure with systems and open architectures that give advanced developer advocate and convey a associate ecosystem with potent third-celebration integrations.”
Symantec Integrates items, functions and companions
Symantec begun constructing ICD two and a half years in the past with its acquisition of Blue Coat methods, which delivered choicest-of-breed net and cloud safety technologies to Symantec’s market-main endpoint, e-mail and facts loss prevention (DLP) technologies. on the time, Symantec saw and heard that valued clientele had been spending great time and substances to combine point technologies in an application to derive rectify cost from their cyber safety investments. So, the business invested in a methodology and roadmap to deliver an built-in platform that enormously reduces can impregnate of operations whereas enhancing the hurry and accuracy of prevention, detection and response in an application to reduce risk.
when you reckon that then, Symantec has:
ICD Platform Earns broad Ecosystem aid
Symantec now has more than a hundred and twenty partners in TIPP, who are edifice or providing greater than 250 current purposes and capabilities that combine with Symantec’s ICD Platform, so business customers can gash back the can impregnate and complexity of their protection operations. in addition, Symantec is now launching a brand current “Innovation Playground” software inside TIPP to simplify integration with startups. the brand current program will allow startup teams to leverage Symantec APIs and capitalize entry to products, engineering materials, and customer innovation days.
“in an application to reduce protection operations complexity and fight these days’s more and more sophisticated adversary, companies exigency products that labor as a platform as a substitute of an army of point items working in silos,” talked about Oliver Friedrichs, VP of security automation and orchestration at Splunk. “Splunk’s aid for the ICD Platform gives their joint shoppers with consolidated views across their safety infrastructure, including incidents flowing from endpoint, internet, network and e mail protection options.”
“At container, safety is a appropriate priority and we're dedicated to presenting their clients with effective controls to offer protection to their sensitive content,” spoke of Niall Wall, senior vice president of companions at field. “Symantec is a founding member of the container believe Ecosystem. we're enthusiastic about Symantec’s integrated Cyber defense and how it will assist our mutual shoppers reduce risk of facts loss, observe advanced threats, and seamlessly compile their security capabilities.”
“protection analysts these days cope with more and more complicated threats, fragmented safety tools, and siloed organizations,” observed wealthy Telljohann, director of business edifice at IBM safety. “To fight this we're considering that the cyber protection landscape is annoying a shift to built-in systems in an application to reduce complexity and price. we absorb developed an integration using Symantec ICD change, so the IBM Resilient Incident Response Platform can provide shrewd orchestration, automation, and enrichment of incidents caused with the aid of Symantec ICD, allowing analysts to reply intelligently to threats.”
ICD Platform Drives consumer Adoption and value
As evidenced by Symantec’s recent quarterly profits, more and more shoppers are stirring beyond particular person products to undertake the ICD platform and portfolio. for example:
“The casual panorama they every pan isn't static and is normally on the flow, as are their purchasers and personnel, so techniques to deal with that contour of environment are very complicated,” observed Emily Heath, vice president and chief suggestions safety officer, United airways. “Visibility of your environment and integration of solutions are a key a portion of that method. for example, if one protection wield catches whatever, it's a lot extra efficient for us if these controls are integrated and may talk seamlessly with each different to assist with precise time detection. moreover, if protection providers create the application to combine across the stack so they don’t must, that effects in a unprejudiced improved result.”
Symantec Extends ICD Platform with current facets & services
Symantec is introducing three current know-how innovations these days that prolong ICD for shared intelligence and shared administration throughout several expertise components, as well as current “hazard aware” information protection capabilities:
For extra guidance on ICD, gladden hunt recommendation from: www.symantec.com/icd.
For greater particulars about TIPP, gladden hunt recommendation from: https://www.symantec.com/expertise-companions.
For extra particulars on how Symantec and companions are leading the shift to integrated Cyber protection, gladden hunt recommendation from: https://www.symantec.com/blogs/feature-experiences/united-we-stand
For greater details on the cyber protection platform shift, gladden hunt recommendation from: https://www.symantec.com/blogs/characteristic-studies/cyber-safety-platform-shift-extra-relaxed-less-complicated
to note an in depth mapping of Symantec product and partner integrations, gladden visit: https://www.symantec.com/integration.
Symantec organisation (SYMC), the realm's main cyber protection company, helps agencies, governments and americans cozy their most necessary information anyplace it lives. companies across the world materialize to Symantec for strategic, integrated options to safeguard in opposition t refined attacks across endpoints, cloud and infrastructure. Likewise, a global group of greater than 50 million people and households weigh on Symantec's Norton and LifeLock product suites to protect their digital lives at domestic and across their contraptions. Symantec operates one of the world's greatest civilian cyber intelligence networks, enabling it to peruse and give protection to against essentially the most superior threats. For more information, gladden talk over with www.symantec.com or connect with us on facebook, Twitter, and LinkedIn.
be alert TO U.S. EDITORS: if you'd fancy additional info on Symantec organisation and its products, gladden hunt recommendation from the Symantec Newsroom at http://www.symantec.com/news.
Symantec and the Symantec brand are emblems or registered logos of Symantec corporation or its associates within the U.S. and other international locations. different names could breathe trademarks of their respective owners.
ahead-searching STATEMENTS: Any ahead-looking indication of plans for items is prefatory and every future unencumber dates are tentative and are discipline to exchange. Any future release of the product or planned changes to product capacity, functionality, or characteristic are subject to ongoing evaluation with the aid of Symantec, and may or can moreover not breathe carried out and may not breathe regarded enterprise commitments by means of Symantec and may no longer breathe relied upon in making paying for selections.
1 source: ESG analysis, casual Detection and Response Survey, December 2018.
2 supply: ESG research record, 2019 technology Spending Intentions Survey, February 2019.
three supply: ESG master Survey outcomes, Cybersecurity panorama: The Evolution of business-type vendors and structures, October 2018.
View source edition on businesswire.com: https://www.businesswire.com/information/domestic/20190227005565/en/
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Toigo Partners International
In the late 1990s, Gal Naor started a company, StorWiz (he later changed the name to StorWize), to foster a technology he developed for delivering data compression at wire speed. Not only was this a frosty understanding and one that struck me as much more efficient than most algorithmic data deduplication approaches in vogue at the time, it had the nifty advantage of being software-based. You could apply the technology to just about any nature of storage to squeeze out the bandwidth required for data transfers and space for storing bits when they came to rest. IBM bought StorWize in 2010, making Naor and his investors rather well to do.
That was the ultimate I had heard of Naor for years. I thought he'd had his fill of storage technology and gotten out of the business just in time to miss the industry consolidation that dramatically reduced the number of vendors in recent years. As it turned out, Naor had "gone quiet" for the past seven years and not just to bask in the afterglow of his success. He was engaged in serious R&D for his next vast adventure. Now, with current technology based on more than 50 patents, Naor has re-entered the fray at the helm of software-defined storage startup StorOne.
His current tech, called TRU Storage, is closer to what they every thought software-defined storage would breathe when the SDS concept was first floated a few years back. The acronym TRU stands for Total Resource Utilization. In this case, it's a designation that's more than aspirational.
I've been acquainting myself with TRU Storage over the ultimate few months. With it, StorOne storage might just set the bar for SDS going forward. (I absorb no stake in the company or contract to promote their wares.)Total Resource Utilization
Gartner reported ultimate summer that SDS and hyper-convergence had reduced the cost per terabyte by increasing the amount of storage a single administrator could manage. However, they had moreover caused a 10% reduce in the utilization efficiency of storage capacity across the enterprise. The judgement for the utilization fail: Storage was being siloed behind different hypervisors in a route that prohibited shared employ by different workloads operating under different hypervisor brands.
With S1, the StorOne TRU Storage product name, neither the workload, nor the hypervisor, nor the hardware used to build the storage infrastructure matters. StorOne storage technology creates universal storage pools that can embrace any storage gear connected via any protocol and enables any workload to employ that resource. Block, file and remonstrate storage can share the very resource, volume and drive. Remarkably, S1 optimizes the overall infrastructure's performance and capacity to more than 90% of advertised peaks. That means you sacrifice fewer IOPS and terabytes in an S1 environment compared with other current approaches, breathe it converged or hyper-converged infrastructure.
StorOne's results absorb yet to breathe evaluated by the Storage Performance Council (SPC) or other benchmarking outfits. Publishing results would likely prove problematic anyway, given the pushback the SPC received when it published the results of DataCore Software testing a brace years ago. fancy StorOne, software-defined storage vendor DataCore took an unconventional path to accelerating I/O -- a software shim that created a parallel I/O processing engine out of unused logical CPU cores, thereby accelerating I/O processing and reducing latency by wonderful levels.
Big box vendors fancy Hewlett Packard Enterprise and IBM thought the resulting comparison unfair, since it demonstrated the very IOPS and latencies could breathe obtained from commodity servers and white box arrays as from highly configured vast iron arrays. Smoke still billows from vendor ears, and a few even offer off-the-record disparagement of SPC methodology to this day.A potential disrupter
StorOne storage is similar to DataCore in terms of its disruptive potential, but it is moreover different. TRU Storage does not employ DRAM caching as DataCore does. It hardly uses caches at all. S1 software runs as a virtual machine under ESXi. It uses a surprisingly low amount of resources in an all-virtual environment to immediately deliver IOPS that exceed the performance-leading all-flash arrays, according to StorOne lab tests.
TRU Storage is closer to what they every thought software-defined storage would breathe when the concept was first floated a few years back.
Recently, StorOne released a test result document, demonstrating 1.7 million IOPS using 24 Western Digital glitter drives (random reads), 15 gigabyte per second sequential reads, 7.5 GBps sequential writes and 10 GBps for mixed workloads of 80/20 reads/writes. By comparison, StoreOne noted, an all-flash array competitor in the market required four times the hardware and expense to deliver only 1.5 million IOPS. For the record, these are internal tests performed by StorOne and not by benchmark benchmark testing outfits.
A persistent integrity guarantee promises full recovery from any hardware or environment failure without data loss. This is enabled by an integrated enterprise data protection and data retention feature that permits zero-time backup and restore and unlimited snapshots with no performance degradation. StorOne storage delivers this functionality without requiring rejiggering RAID levels on connected equipment, adding remembrance or processor or changing configurations.
S1 is quite literally plug-and-play.
Perhaps the most disruptive feature of StorOne storage is its cost tag. Naor says S1 will breathe offered as software, as a service or bundled with white box arrays for the cost of a penny per gigabyte from the software-defined storage vendor. That is lower than most cloud storage offerings, every of which don't offer the performance characteristics or integrity guarantee of S1.
I'll breathe watching to note how the tech plays out at scale and in geographically distributed installations, including hybrid cloud on-premises infrastructure. When I bring this up, Naor simply smiles his "just stand by and breathe patient" smile, as he insists TRU Storage technology will scale without problems. Given his record of technological success, I'm willing to give him the capitalize of the doubt.
Coitally delivered microbicide gels containing antiretroviral drugs are necessary for HIV prevention. However, to date, microbicides absorb contained entry or transpose transcriptase inhibitors that screen early steps in virus infection and thus exigency to breathe given as a preexposure dose that interferes with sexual practices and may circumscribe compliance. Integrase inhibitors screen late steps after virus infection and therefore are more suitable for post-coital dosing. They first determined the kinetics of strand transfer in vitro and confirmed that integration begins about 6 hours after infection. They then used a repeat-challenge macaque model to assess efficacy of vaginal gels containing integrase strand transfer inhibitors when applied before or after simian/human immunodeficiency virus (SHIV) challenge. They showed that gel containing the strand transfer inhibitor L-870812 protected two of three macaques when applied 30 min before SHIV challenge. They next evaluated the efficacy of 1% raltegravir gel and demonstrated its skill to protect macaques when applied 3 hours after SHIV exposure (five of six protected; P < 0.05, Fisher’s exact test). Breakthrough infections showed no evidence of drug resistance in plasma or vaginal secretions despite continued gel dosing after infection. They documented rapid vaginal absorption reflecting a short pharmacological lag time and preeminent that vaginal, but not plasma, virus load was substantially reduced in the breakthrough infection after raltegravir gel treatment. They provide a proof of concept that topically applied integrase inhibitors protect against vaginal SHIV infection when administered shortly before or 3 hours after virus exposure.INTRODUCTION
HIV-1 continues to spread globally, highlighting the exigency to develop efficacious biomedical interventions to circumscribe its transmission. In addition to vaccine development, there are several antiretroviral (ARV) drug–based intervention strategies currently being advanced, including oral preexposure prophylaxis (PrEP) treatment as prevention as well as topical PrEP with gels or other ARV delivery methods such as intravaginal rings and tablets (1–3). ARV gels absorb several advantages for HIV prevention. When applied to the mucosal site of virus exposure, gels can rapidly dose tissues with drug concentrations that are much higher than those achieved by oral dosing and with minimal systemic drug exposures and drug toxicities (3–9). Topical gels are user-controlled and can breathe optimally formulated with a single drug or in combination for both vaginal and rectal application (4–6, 10).
The CAPRISA 004 crucible evaluated the safety and effectiveness of a vaginal gel containing 1% tenofovir (TFV) and provided the first evidence of efficacy by this intervention strategy. The study demonstrated a 39% reduction of HIV acquisition in women who applied the gel pericoitally (1). Effectiveness was relative on adherence and was establish to enlarge to 54% in women who reported frequent gel employ (>80% adherence) compared to only 28% in women who were <50% adherent. A subsequent study (VOICE) evaluating the very gel formulation in a noncoital daily dosing modality was stopped because of the absence of protection (2). TFV testing in plasma revealed poor adherence among participants, likely explaining the exigency of efficacy (3). These data and similar findings from oral PrEP studies highlight the challenges of identifying intervention strategies that absorb both towering biological efficacy and enhanced user compliance (4).
Coitally used topical gels are necessary HIV prevention methods. However, to date, every such products reckon on a preexposure dose that can interfere with sexual practices, and may breathe limited by partner’s acceptance and the exigency to anticipate sex. They reasoned that a topical product that can breathe applied by women after sex will absorb less impact on sexual practices, absorb better user control, and circumvent the exigency for sex anticipation and partner acceptance. These advantages can every potentially enhance compliance and effectiveness. However, every topical ARV-based gels in progress employ entry or transpose transcriptase (RT) inhibitors that screen early steps in the virus infection cycle, which raises questions about their suitability for postexposure prophylaxis (PEP) (1, 5–7). Macaque models of rectal or vaginal transmission of the simian immunodeficiency virus (SIV) or the chimeric simian/human immunodeficiency virus (SHIV) absorb provided necessary data confirming the efficacy of topical gels with RT inhibitors such as TFV and MIV150 when applied before virus challenge, but demonstrated loss or substantial reduction in efficacy when applied 1 to 2 hours after virus challenge (8–10). These data likely reflect the exigency to adequately dose mucosal tissues with RT inhibitors before virus exposure and insinuate that drugs from different classes that target later steps in virus infection may breathe more efficacious for PEP.
HIV or SIV integration into the host DNA is the ultimate step before an irreversible infection of a cell takes place. Integration is a multistep process catalyzed by the viral integrase that follows transpose transcription and starts in the cytoplasm with 3′-end processing of viral DNA, continues with translocation of the processed DNA to the nucleus, and is completed by integrating the viral DNA into the host chromosome by a strand transfer mechanism (11, 12). The integration process can breathe inhibited at various stages by three main classes of integrase inhibitors: 3′-processing inhibitors (13), strand transfer inhibitors (14), and macromolecular complicated LEDGF/p75-IN inhibitors (15). Strand transfer inhibitors are the most developed and absorb been validated as potent inhibitors of HIV replication in vitro, ex vivo, in animal studies, and in clinical trials (16, 17). The mechanism of action of strand transfer inhibitors is characterized by their skill to chelate the Mg2+ ions in the catalytic core domain of integrase, thereby disrupting the interaction between the integrase/viral complementary DNA preintegration complicated and the target DNA (18). The only clinically approved integrase inhibitors licensed for the treatment of HIV-infected persons are raltegravir (RAL), elvitegravir, and dolutegravir, every of which are strand transfer inhibitors (14, 19). The compound L-870812 (L-812) is a potent investigational strand transfer inhibitor and has been shown to breathe effectual in reducing SIV replication in macaques (16). For PEP, blocking the ultimate step in virus integration by strand transfer inhibitors can breathe advantageous because it can extend the post-coital dosing window (14, 20). RAL and L-812 are equally potent on both HIV and SIV integrase, with IC50 (50% inhibitory concentration) in the nanomolar scope (1 to 4 nM), which provides an occasion to test in macaque models the concept of topical prophylaxis by this class of drugs for both pre- and postexposure protection (16, 17).
Here, they used a reiterate low-dose vaginal challenge macaque model to evaluate the efficacy of gels containing L-812 or RAL administered before or after virus challenge, respectively. This well-established macaque model has several advantages, including the employ of pigtailed macaques, which absorb a menstrual cycle and vaginal anatomy similar to women, an inoculum dose with viral RNA levels in the scope similar to those detected in seminal fluid, twice-weekly virus challenges to mimic high-risk human exposure, and an SHIVSF162p3 inoculum that uses an R5-tropic envelope similar to that of most transmitted HIV (10, 21, 22). The repeated challenges in this model absorb the added advantage of increasing statistical power without requiring great groups of animals because they allow measurement of protection over more than one transmission event per animal. They moreover performed time-of-drug addition studies in vitro to define the window for strand transfer inhibition and to better inform in vivo dosing.RESULTS Gel formulation and impact of vaginal fluids on antiviral activity
L-812 was successfully formulated in a hydroxyethyl cellulose (HEC) gel at a maximum solubility of 2.3 mg/ml (0.23%). Because of higher water solubility, RAL was formulated in a HEC gel at a concentration of 10 mg/ml (1%). Both gels were clear, viscous, and free of particulates and remained highly energetic for up to 1 year at latitude temperature. To investigate whether biological factors present in the vagina, including mucus, enzymes, and microflora, may compromise the stability and potency of RAL gel, they assessed the antiviral activity of RAL gel incubated in cervicovaginal fluids (CVFs). motif 1 shows that CVF had no impact on RAL activity because no contrast in IC50 was observed between RAL incubated in CVF (IC50 = 2.6 ± 1.1 nM) or saline (1.7 ± 1.2 nM).Fig. 1. Antiviral activity of RAL gel is maintained in the presence of vaginal fluids.
A TZM-bl reporter cell line was used to evaluate antiviral activity (IC50) of RAL gel preincubated (24 hours) in saline buffer or CVF. Data are means ± SE of three independent experiments.Time-of-drug addition reveals a wide window for inhibition by integrase inhibitors
To better define the timing of transpose transcription and integration and to inform the dosing modality in vivo, they performed time-of-drug addition experiments using single-cycle infections of TZM-bl cells with vesicular stomatitis virus (VSV)–pseudotyped HIV-1. The kinetics of postexposure inhibition by the RT inhibitor TFV was compared with that of RAL. motif 2 shows that TFV maintained towering (>95%) protection up to 2 hours after infection, but was only ~50% protective when added at 5 hours after infection. In contrast, RAL provided towering protection (>90%) at 6 hours and remained above 50% protection for up to 10 hours after infection. These data insinuate that strand transfer is delayed by at least 4 hours after transpose transcription, providing a longer window to screen infection with integrase inhibitors than inhibitors of transpose transcription.Fig. 2. Kinetics of the window of inhibition by RT and integrase inhibitors.
Protection of TZM-bl cells from infection with VSV-HIV was evaluated after addition of the RT inhibitor TFV or the integrase inhibitor RAL at the indicated times after infection. Data are means ± SE of three independent experiments.Preexposure protection by a vaginal integrase inhibitor gel
We first evaluated whether topically applied integrase inhibitors administered before virus challenge protected macaques from SHIV infection. Three pigtailed macaques received vaginal gel containing 0.23% L-812 30 min before vaginal SHIVSF162p3 exposure. Infection outcome after 14 twice-weekly vaginal challenges was evaluated against 10 untreated controls receiving placebo gel (1 real-time and 9 historic controls). every controls received the very placebo gel and were challenged with the very virus stock under identical experimental conditions. In this model, pigtailed macaques are more susceptible to infection during the late luteal aspect of the menstrual cycle (10, 23). As previously described, the risk for infection was calculated per month or menstrual cycle (10). motif 3A shows the infection rates in animals treated with L-812 and placebo gel. Nine of 10 controls (90%) became infected after 14 challenges over a 2-month period; seven, including the real-time control, were infected after eight challenges (1 month), and two were infected during the second month. In contrast, two of the three macaques treated with L-812 gel remained uninfected after 14 challenges (estimated efficacy = 76%; P = 0.057, Fisher’s exact test); the breakthrough infection occurred after nine challenges (week 5), whereas both protected animals remained seronegative and had undetectable SHIV sequences in plasma or PBMCs throughout the 10-week follow-up period. L-812 gel efficacy was estimated at 76% on the basis of 9 of 13 infections per months (or cycles) at risk among controls and 1 of 6 for treated animals.Fig. 3. Protection by PrEP and PEP with vaginal gels containing integrase inhibitors.
Survival curves represent the cumulative percentage of uninfected macaques as a duty of the number of months at risk for infection during the study period (eight challenges per month). (A) Gel containing L-812 (solid line) or placebo (dotted line) was applied vaginally 30 min before each SHIV exposure (up to 14 SHIV exposures). Controls embrace one real-time (solid circle) and nine historic controls. (B) Gel containing RAL (solid line) or placebo (dotted line) was applied vaginally 3 hours after SHIV exposure (up to 20 SHIV exposures).Postexposure protection by RAL gel
We next explored the efficacy of 1% RAL gel applied 3 hours after vaginal SHIV exposure. The 3-hour dosing interval falls within the dosing window for maximal in vitro inhibition of strand transfer and is behaviorally feasible for post-coital application. motif 3B shows infection outcomes in macaques that were challenged twice weekly for 2.5 months (about two menstrual cycles) with SHIVSF162p3 and administered placebo (n = 4) or 1% RAL (n = 6) gel 3 hours after each virus challenge. every four macaques that received placebo gel became infected by 10 weeks (20 challenges). In contrast, five of six macaques treated with RAL gel 3 hours after SHIV exposure remained uninfected after 20 SHIV exposures and the 10-week follow-up period. every infected macaques, including the breakthrough infection, exhibited typical SHIV RNA viremia and detectable proviral DNA in PBMCs and seroconverted 1 to 3 weeks after first detectable plasma SHIV RNA was observed. Risk for infection differed by study group (P < 0.05, Fisher’s exact test), with RAL gel efficacy estimated at 84% on the basis of 4 of 7 infections per months (or cycles) at risk among controls and 1 of 11 for treated animals. Together, these data insinuate that topically applied integrase inhibitors can obviate vaginal infection when administered shortly before or up to 3 hours after SHIV exposure.Systemic drug concentrations after vaginal gel dosing
Measuring drug exposure in blood is critical to understanding kinetics of drug release and systemic absorption after vaginal gel dosing. Systemic drug exposures are moreover necessary for determining impact on acute viremia and emergence of drug resistance in animals that fail gel prophylaxis and continue receiving gel treatment. They measured L-812 and RAL concentrations in plasma 30 min after each gel administration to determine systemic drug exposure at time of challenge and moreover assessed the impact of the menstrual cycle on vaginal drug absorption. They absorb previously preeminent with gels containing TFV or emtricitabine (FTC) that systemic drug exposure peaks within 1 hour after gel dosing and is highly influenced by the menstrual cycle with higher drug concentrations in plasma observed during the progesterone-dominated luteal aspect when the vaginal epithelium is thin (24, 25). They analyzed longitudinally both progesterone and drug concentrations in plasma in every macaques that were treated with L-812 or RAL gel (Fig. 4, A and B). With the exception of one (PMl), every macaques experienced at least two menstrual cycles during the study period. Plasma drug concentrations 30 min after vaginal dosing showed similar trends in every animals with substantially higher concentrations consistently detected after peak progesterone. The express concentration of L-812 and RAL 1 week after peak progesterone (late luteal phase) was 115.3 ± 20.8 ng/ml (range, 74 to 140) and 111 ± 31.3 ng/ml (range, 0 to 260) compared to only 1.5 ± 0.5 ng/ml (range, 0 to 6) and 5.8 ± 4.9 ng/ml (range, 0 to 39.8) 1 week before peak progesterone (late follicular phase). The data authenticate that both drugs were rapidly released from the HEC-based gel formulation. The longitudinal analysis of plasma drug concentrations in the two breakthrough infections (PBg-2 and PSz-1) revealed similar peak drug levels and absorption trends to animals that were protected. PSz-1 had detectable RAL in plasma at the estimated time of infection, 7 days (226 ng/ml) to 10 days (259 ng/ml) before the first detectable SHIV RNA. However, PBg-2 had undetectable L-812 at day 7 and 17 ng/ml at day 10. Although limited to only two animals, these results conclude not demonstrate a transparent association between drug concentrations in plasma and protection.Fig. 4. Longitudinal analysis of plasma drug and progesterone concentrations in pigtailed macaques dosed with vaginal gels containing integrase inhibitors.
(A) Three macaques were dosed twice weekly with L-812 gel (dotted line); shaded areas demonstrate progesterone concentrations. (B) Six macaques dosed twice weekly with RAL gel (dotted line); shaded areas demonstrate progesterone concentrations. every drug concentrations were analyzed from plasma collected 30 min after each vaginal gel application. SHIV+ denotes first detectable SHIV RNA in plasma.Virus replication in breakthrough infections
We next explored the impact of continued vaginal gel dosing on plasma and vaginal virus shedding in macaques PBg-2 and PSz-1, which became infected despite receiving L-812 or RAL gel, respectively. Infected macaques continued to receive twice-weekly placebo (n = 5), L-812 (n = 1), or RAL (n = 1) gel treatment for up to 8 weeks (16 gel applications) after the first SHIV RNA was detected in plasma. Plasma SHIV RNA detected in both breakthrough infections was evaluated against the five real-time controls. Virus shedding was assessed only in the RAL failure and controls receiving placebo gel. motif 5A shows the virus load kinetics in both breakthrough infections compared to controls. The peak viremia in PBg-2 (6.08 log10 RNA copies/ml) and PSz-1 (6.7 log10 RNA copies/ml) was similar to the median peak viremia of the five untreated controls (7.01 ± 0.98 log10 RNA copies/ml), showing no contrast in acute viremia (P = 0.29, Wilcoxon rank sum test). In contrast, Fig. 5B reveals that the peak vaginal SHIV RNA (3.5 log10 copies/ml) and the frequency (4 of 15) of SHIV RNA detected in vaginal fluids were both significantly lower in PSz-1 than in placebo-treated animals that had a median SHIV RNA peak of 5.2 log10 ± 0.42 copies/ml (P = 0.007) and frequency of 36 of 60 (P < 0.0001), respectively. The reduction in vaginal SHIV shedding could reflect the antiviral activity by RAL gel.Fig. 5. Plasma and vaginal virus loads in macaques with breakthrough infection and controls.
(A) Individual virus load kinetics in breakthrough infections under continued twice-weekly dosing with gel containing L-812 (diamonds) or RAL (circles) compared to median virus loads of controls (n = 5, dotted black line). Time 0 indicates first detection of SHIV in plasma. (B) Individual virus load kinetics in vaginal secretions collected twice weekly before dosing with gel containing RAL (solid line; great circles) or placebo (n = 4, black dotted lines). Filled symbols bespeak samples sequenced for drug resistance in plasma and vaginal fluids. The broken line denotes the circumscribe of quantitation of the virus load assay (50 copies/ml).No evidence of emerging drug resistance after continued gel dosing in breakthrough infections
Because L-812 and RAL were detected in plasma in the breakthrough infections, it was necessary to determine whether these systemic drug exposures led to drug resistance. Sequence analysis of the integrase region spanning the N-terminal and core domain (amino acids 1 to 234) in virus first detected in plasma showed that both breakthrough infections were initiated with wild-type virus (fig. S1). Plasma specimens collected at peak viremia and up to 8 weeks after infection every revealed wild-type genotypes (Fig. 5A). To examine drug resistance emergence at the site of gel application, they moreover genotyped SHIV RNA isolated from vaginal secretions in the RAL breakthrough infection (fig. S1). Vaginal fluids with detectable SHIV (n = 4) were genotyped, and every establish to breathe wild nature (Fig. 5B). Thus, despite twice-weekly vaginal dosing for 8 weeks after infection, no evidence of drug resistance in plasma or genital secretions was observed (fig. S1).DISCUSSION
This study provides a proof of concept that topically applied strand transfer integrase inhibitors protect macaques against repeated vaginal SHIV challenges. Protection was observed when the vaginal gel was applied shortly before and, more importantly, 3 hours after virus challenge. The protection by postexposure dosing reflects the advantages of the late-acting strand transfer inhibitors that are needed several hours after virus entry, thus providing an optimal window for post-coital dosing that was previously not feasible with entry or RT inhibitors (8, 9). Because applying gel after sex may interfere less with sexual practices, their data advocate exploring current post-coital modalities with integrase inhibitors for enhanced acceptance and compliance in women.
We used single-round infections to determine the kinetics of transpose transcription and integration and demonstrate that in the HeLa-derived TZM-bl cells, strand transfer starts more than 6 hours after infection, consistent with previous findings in different cell infection systems (21, 22, 26, 27). The integration step may retract longer in less activated primary cells that are commonly present in vivo in which transpose transcription and integration proceed more slowly than in transformed cell lines (26, 28). Thus, it is likely that the dosing window in vivo may breathe longer than the 3 hours evaluated in this study. Integrase inhibitors, including RAL, are known to bind tightly to preintegration complexes with dissociation half-lives of >7 hours, which ensures longer antiviral activity in infected cells even if tissue drug concentrations subsequently putrefy to suboptimal levels (19). They moreover note the detection of drug in plasma within 30 min of vaginal gel dosing, which reflects rapid absorption through vaginal tissues. The rapid absorption of RAL and the exigency of intracellular activation insinuate that the pharmacological lag time for in vivo activity may breathe minimal. Additional macaque studies of longer postexposure gel applications are needed to further define the post-coital window of protection by strand transfer inhibitors.
We moreover document in pigtailed macaques changes in plasma drug levels during the menstrual cycle and commemorate higher concentrations during the progesterone-dominated luteal phase. These results reflect a higher vaginal permeability peaking during the luteal aspect likely due to thinner epithelium, increased porosity, changes in mucus composition, and other factors (23, 24, 29). The influence of the menstrual cycle on vaginal drug absorption moreover explains the cyclical changes in plasma concentrations of TFV and FTC they absorb previously observed in pigtailed macaques dosed with gels containing TFV and FTC (6). They moreover examined the relationship between plasma drug concentrations and vaginal efficacy because Cranage et al. previously establish a positive association between TFV concentration in plasma and the degree of rectal protection by rectal TFV gel (9). With only two breakthrough infections, they were unable to discern any trend because one animal had towering drug concentrations around the estimated time of infection and one did not. These results likely highlight the added complexity in identifying pharmacological markers of vaginal protection relative to rectal protection and the confounding role of the menstrual cycle modulating both susceptibility to infection and drug permeability (23). Nevertheless, the data demonstrate the usefulness of the pigtailed macaque model that has a menstrual cycle similar to that of women, and advocate similar evaluations in women to better define the impact of menstrual cycle and hormonal contraceptives on vaginal pharmacokinetic parameters and efficacy.
Both L-812 and RAL are potent drugs with IC50 in nanomoles; however, they establish that the concentrations in gels that conferred high, yet incomplete, in vivo protection were in the micromolar range, or about 1 million–fold higher than those concentrations needed for in vitro inhibition. The higher drug concentrations required for in vivo efficacy absorb been consistently observed with other gels containing entry or RT inhibitors and likely reflect the distribution kinetics of the drug through the mucosa to achieve concentrations needed to protect the great surface locality of the vagina for a significant period of time (6, 7, 10). Previous labor with TFV gels in macaques has shown that the pharmacologically energetic TFV-diphosphate (TFV-DP) concentrations in vaginal lymphocytes prognosticate in vivo efficacy, and topical dosing by gel or intravaginal ring that achieved TFV-DP concentrations exceeding the in vitro IC95 values in these lymphocytes was associated with complete protection (10, 30). Thus, it is tempting to surmise that a similar pharmacological correlate will moreover apply to RAL. However, confirming this correlate will require measuring intracellular RAL concentrations in vaginal lymphocytes, which could breathe challenging because unlike intracellularly bound TFV-DP, RAL may diffuse out during the cell purification process as extracellular RAL concentrations decrease. Thus, alternative tissue testing methods are needed.
The potential for drug resistance to develop when prophylaxis fails raises concerns, especially with drugs that are moreover used for treatment. Prolonged systemic treatment of SHIV-infected macaques with L-812 has been establish to select drug-resistant variants with the integrase mutation (N155H) (20). They demonstrate no evidence of drug resistance emergence in plasma from either the L-812 or RAL breakthrough infection despite continued twice-weekly gel dosing for up to 8 weeks after infection. Although larger numbers of animals are needed to define incidence of resistance in animals failing prophylaxis, the finding of wild-type SHIV suggests that both gel failures are not due to a selection of a low-frequency drug-resistant variant in the challenge virus, and likely reflects insufficient selective pressure by the systemically absorbed drug from the twice-weekly gels. These systemic drug exposures are much lower than those observed from oral dosing and likely contribute to lower risks of drug resistance (20). Indeed, they establish no reduction in the acute viremia in both breakthrough infections compared to controls, further supporting the minimal systemic antiviral activity. These findings are similar to TFV gel breakthrough infections but vary from those in macaques that fail oral PrEP in which reductions in virus loads and selection of drug-resistant variants absorb been observed (7, 9). Given that the antiviral activity from gels remains largely concentrated in the vaginal compartment, they moreover evaluated the viral population in the genital secretions from the RAL breakthrough infection. They detected only wild-type SHIV, but preeminent that, in contrast to plasma virus load, vaginal virus shedding was substantially reduced in this macaque. The suppression of virus shedding may reflect the antiviral activity from gel dosing, which may moreover absorb reduced drug resistance selection. These data are only from one animal and require confirmation, but they conclude point to broader implications because reductions in vaginal virus shedding during acute infection may lower transmissibility.
Our study has several limitations. The virus challenges did not embrace cell-associated virus and were done in the absence of semen or semen-derived factors shown to enhance HIV infection in vitro (18). However, recent data insinuate tiny impact of semen on vaginal SIV transmission in rhesus macaques (31). Also, SHIV exposures were made on intact vaginal mucosa without trauma associated with coitus, concurrent genital ulcers, or bacterial vaginosis, every of which can enlarge the risk of HIV acquisition (26, 27). As previously noted, only two breakthrough infections were available to study vaginal SHIV shedding and drug resistance, and thus, these observations require confirmation in larger numbers of animals.
In conclusion, they demonstrate preclinical in vivo data that advocate the employ of topical integrase inhibitors for HIV prevention. They highlight the advantage of using this class of drugs in current post-coital modalities that were previously not possible with entry or RT inhibitors. This study supports evaluation of this class of drug for HIV prevention, including minuscule molecules that conclude not overlap with clinically approved integrase inhibitors in optimized gels or other dosage forms such as vaginal rings or tablets (28, 32, 33). The study moreover supports further evaluation of post-coital modalities in women for enhanced acceptability and compliance.MATERIALS AND METHODS Gel formulations
L-812 and RAL were provided by Merck and the National Institutes of Health (NIH) AIDS Research and Reference Reagent Program, respectively. L-812 was formulated at 0.23% (w/w) in a 2% HEC gel containing 20% propylene glycol, 0.02% propylparaben, 0.18% methylparaben, and 0.02% EDTA. RAL was formulated at 1% (w/w) in a 2% HEC gel containing 0.017 M phosphate-buffered saline (PBS) (pH 7.4), 0.02% propylparaben, 0.18% methylparaben, and 0.02% EDTA. A 2% HEC gel formulated without drug was used as a placebo control. Gels were stored at latitude temperature, and antiviral activity was confirmed throughout the course of the study with a TZM-bl reporter assay to measure drug susceptibility.Virus stock
SHIVSF162P3 (SIVmac239 backbone with an HIV-1 subtype B, CCR5-tropic envelope) was obtained from the NIH AIDS Research and Reference Reagent Program and was propagated in peripheral blood mononuclear cells (PBMCs) from pigtailed macaques as previously described (34, 35). The virus stock titer was calculated in pigtailed macaque PBMCs and diluted to the challenge dose of 10 TCID50 (median tissue culture infectious dose) (1.5 × 106 RNA copies per exposure).Antiviral activity and stability of RAL gel
TZM-bl cells (NIH AIDS Research and Reference Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases, NIH) were used to determine the IC50 of gels containing integrase inhibitors, and data were analyzed by GraphPad Prism (v5.02) software. For stability testing, 1% RAL gel (20 mM) was diluted 10-fold in PBS or macaque CVF and incubated overnight at 37°C. RAL-saline and RAL-CVF were then serially diluted (20-0 μM) in complete medium, and 100 μl was added to wells of a 96-well plate in triplicate. Complete medium (50 μl) containing TZM-bl cells (3 × 106/ml) was plated in each well. After 15 min of incubation, 50 μl of SHIVSF162p3 stock, diluted to a final concentration resulting in a signal of about 105 relative light units, was added to each well. After 48 hours of incubation at 37°C, 100 μl of medium was removed and replaced with 100 μl of Bright-Glo (Promega Corp.), and the luminescence was measured with PerkinElmer Victor X2 Multilabel Plate Reader. Inhibition was determined on the basis of reduction from the virus-only control and presented as the percentage of cells protected (mean ± SEM).Kinetics of postexposure inhibition
Time-of-drug addition experiments were carried out with single-cycle infections to determine the inhibition window for RT and integrase inhibitors. TFV or RAL was added to infected TZM-bl cells at concentrations exceeding IC99 (200 μM TFV or 20 μM RAL) at different times after infection (0 to 24 hours). Briefly, TZM-bl cells were preincubated with VSV-pseudotyped HIV-1 NL4-3-ΔE-EGFP (VSV-HIV) for 1 hour to establish infection. A VSV-pseudotyped virus was used to ensure that every reported infectivity was derived from a single round of infection. Cells were washed twice with PBS and treated with 0.25% trypsin (Cellgro) to remove every cell-bound and cell-free viruses. Infected cells were resuspended in complete medium and plated (3 × 104 per well) in triplicates in a 96-well plate. Drug was added at 1-hour time increments up to 24 hours. The percentage of cells protected at each time point was determined and plotted as % cells protected ± SEM relative to time when drug was added after infection.Efficacy studies
The efficacy of integrase inhibitors against vaginal transmission was evaluated in female pigtailed macaques under conditions similar to those described in other studies (6, 10). Macaques were administered gel (3 ml) vaginally and challenged with a low-dose SHIVSF162P3 inoculum (10 TCID50) twice weekly (every 3 to 4 days) for up to 10 weeks (about two menstrual cycles). every animals were anesthetized and remained recumbent for 30 min after each gel application and 15 min after virus inoculation to minimize leakage. For preexposure efficacy measurements, macaques received placebo (n = 1) or 0.23% L-812 (n = 3) gel vaginally 30 min before each vaginal SHIV exposure (up to 14 challenges). For postexposure efficacy, anesthetized animals were challenged with SHIV (up to 20 challenges) and remained incumbent for 15 min after receiving the virus to minimize leakage and then returned to their cages. After 3 hours, macaques were anesthetized and administered 3 ml of placebo (n = 4) or 1% RAL (n = 6) gel. For each study, blood was collected 30 min after gel dosing to monitor for drug levels and SHIV infection. Virus challenges were stopped when a macaque became SHIV RNA-positive (SHIV+) in plasma. every infected macaques continued to receive study gel (placebo, L-812, or RAL) twice weekly for up to 8 weeks after infection. every experiments were done under highly controlled conditions by the very personnel with the very virus stock, inoculum dose, and procedures as described in previous studies (6). These studies adhered to the pilot for the reliance and employ of Laboratory Animals (Institute for Laboratory Animal Research, 1996); every procedures were approved by the Institutional Animal reliance and employ Committees of both the Centers for Disease Control and Prevention (CDC) and the Yerkes National Primate Research Center, Emory University.Monitoring systemic infection, virus shedding, and drug resistance
Systemic infection was monitored twice weekly by screening for SHIV RNA in plasma with a quantitative real-time transpose transcription polymerase chain reaction (PCR) assay with a sensitivity of 50 RNA copies/ml, as previously described (6, 34, 36). The estimated time of infection was defined as 7 to 10 days before the first SHIV-positive specimen to account for the eclipse period between infection and detection of SHIV RNA in plasma. Infected macaques were monitored for emergence of drug resistance in both plasma and CVF by benchmark sequence analysis of SIVmac239 integrase reference sequence ground pairs 3624 to 4332 (amino acids 1 to 234). CVF specimens were collected by instilling and recollecting about 5 ml of infertile PBS in the vaginal cavity. CVFs were centrifuged for 15 min at 400g to pellet cells and debris. Virion-associated RNA was extracted from CVF supernatant (1 ml) with a Qiagen viral RNA kit. PCR amplification of PBMC proviral DNA was done with primers and probes specific for SIVmac239 pol (6, 34). Serologic testing was performed with a synthetic peptide enzyme immunoassay (Genetic Systems HIV-1/HIV-2 Plus O; Bio-Rad). Animals were considered protected if they tested negative for SHIV RNA in plasma and SHIV DNA in PBMCs and remained seronegative during the challenge period of the study and the 10-week follow-up in the absence of challenge and gel application.Measurement of drug concentrations in plasma
L-812 and RAL levels in plasma were measured in macaques 30 min after vaginal administration of 0.2% L-812 and 1% RAL gel, respectively. Briefly, drug was extracted from 100 μl of plasma by protein precipitation with 350 μl of methanol containing 200 ng of the unstudied analyte as the internal benchmark (RAL was used as an internal benchmark for L-812 and vice versa). Supernatant containing the drug from precipitation was evaporated to near dryness under vacuum and then resuspended in high-performance liquid chromatography (HPLC) buffer A (9.9 mM acetic acid, 5.9 mM ammonium hydroxide, and 9.4 mM formic acid in H2O). Drug concentrations were analyzed by HPLC–tandem mass spectrometry system with a Shimadzu Prominence HPLC (Shimadzu Scientific) system equipped with a 150 × 2.0–mm Ascentis Phenyl column (Sigma-Aldrich) and an AB Sciex 3200 QTRAP mass spectrometer. Chromatography was done with a linear gradient of 20 to 90% buffer B (acetonitrile containing 9.9 mM acetic acid, 5.9 mM ammonium hydroxide, and 9.4 mM formic acid). Mass spectrometer was hasten in positive MRM (multiple reaction monitoring) mode, and the following transitions were monitored: for RAL, mass/charge ratio (m/z) 445.2/109.0 and 445.2/361.5, and for L-812, m/z 354.1/109.0 and 354.1/203.0. The assay had a lower circumscribe of quantification of 5 ng/ml for both analytes and benchmark curve R2 values greater than 0.99.Menstrual cycle and drug absorption monitoring
A longitudinal assessment of drug absorption in relation to menstrual cycle was done by measuring plasma drug and progesterone levels 30 min after vaginal gel application throughout the challenge period for each animal and each cycle. Plasma samples collected twice per week (every 3 to 4 days) were analyzed for drug levels as described above, and progesterone levels were measured at the University of Wisconsin National Primate Research hub with an enzyme immunoassay (37).Statistical methods
Fisher’s exact test was used to compare the number of infections per number of months at risk by study group. Risk for infection per month, or approximately per menstrual cycle, with biweekly challenge doses throughout each month, considers that cycling pigtailed macaques are more susceptible to infection during the luteal aspect of the menstrual cycle as previously demonstrated in this model (23). Intervention efficacy was calculated as 1 − (p1/p0), where p1 and p0 denote the harmony of months with incident infection for intervention and control animals, respectively (38). SAS version 9.2.1 or GraphPad Prism 5 for Windows (GraphPad Software, www.graphpad.com) was used for every statistical analyses.REFERENCES AND NOTES
The VOICE Study (MTN-003) (2013).
J. Marrazzo, G. Ramjee, G. Nair, T. Palanee, B. Mkhize, C. Nakabiito, M. Taljaard, J. Piper, K. Gomez, M. Chirenje, Pre-exposure prophylaxis for HIV in women: Daily oral tenofovir, oral tenofovir/emtricitabine, or vaginal tenofovir gel in the VOICE Study (MTN 003), paper presented at the 20th Conference on Retroviruses and Opportunistic Infections, Atlanta, GA, 3 to 6 March 2013.
C. Dobard, S. Sharma, impact of Depo-Provera and menstrual cycle on vaginal absorption of antiretroviral drugs from gels in pigtail macaques, paper presented at the 20th Conference on Retroviruses and Opportunistic Infections, Atlanta, GA, 3 to 6 March 2013.
Good day, CIOs. Estonia’s tech-savvy population has helped transform the country into an innovation hub, with the tiny Baltic nation hitting above its weight in launching startups. Similarly, Estonia’s geographical location has propelled the government’s approach to technology. Estonia is a leader in cybersecurity, but also blockchain and e-citizenship, a digital-first approach to sovereignty that makes sense when Russia is your neighbor. Now Estonia is working toward a national policy concerning artificial intelligence. CIO Journal’s Sara Castellanos talks with Siim Sikkut, Estonia’s chief information officer, about the effort.
A national AI action plan. “We could just tinker around without having a coordinated, coherent plan,” said Mr. Sikkut. “But if you want to create a concentrated application and prance tough and fast, you exigency to motif out what to apportion in motion, (what) barriers to break, and that’s what the system gives us.” Estonia’s upcoming action system will address issues such as the extent to which government should employ AI without being overly reliant on the technology or intruding on privacy, Mr. Sikkut said.
A minuscule club. The country, with a population of about 1.3 million, will unite several others that absorb developed national strategies to promote the employ and progress of AI, including China, Finland, Canada, France and Germany. Guidelines for how to deploy AI, and to what extent, will breathe critical for governments seeking to maximize their relevance in the current global economy, said Keith Strier, EY's global and Americas artificial intelligence leader and the lead partner and senior advisor on the project with Estonia. “Not having a national AI action system will express a failure to really coordinated focused resources, which, in a smaller country is more essential because there’s more limited resources.”
What happens in Vegas (goes to the cloud). Amazon.com Inc.'s cloud unit unveiled a number of goodies at its annual event in Las Vegas, Among them:
VMware-enabled hardware for hybrid cloud users to employ in their own data centers through a service called AWS Outposts. (GeekWire).
Its own server chips, called Graviton, built on the ARM architecture used in smartphone chips. (Wired), and a machine learning chip, Inferentia. (Reuters)
A managed blockchain offering and Amazon Quantum Ledger Database, "where customers can replicate a copy of their blockchain network activity." (CNBC)
A Windows-compatible service targeting potential Azure users. (Bloomberg)
A remote-controlled car that can breathe trained using re-enforced learning. (Business Insider)
Facebook emails expose discussion on monetizing user data. Internal emails, most from 2012 to 2104, demonstrate that Facebook Inc. considered charging companies for continued access to user data several years ago. The emails moreover bespeak that Facebook employees discussed pushing some advertisers to spend more in recur for increased access to user information. Writes Journal's Deepa Seetharaman: "Taken together, the internal emails demonstrate the company discussing how to monetize its user data in ways that are employed by some other tech firms but that Facebook has said it doesn’t do."
Amazon makes inroads selling medical supplies to the sick. A growing number of doctors around the U.S. can direct a patient to Amazon.com Inc. for medical products via an app embedded in the patient’s private medical record, the WSJ's Melanie Evans reports. Hospitals bid the goal is to supplant the handwritten shopping lists doctors often hand people. But privacy experts express concern that patients could unwittingly share personal and potentially sensitive health information with Amazon. Federal health-privacy law, enacted in 1996, was never intended to address patients unwittingly sharing sensitive medical data while shopping online, privacy lawyers say.
Microsoft wins $480 million AR Army contract. Microsoft Corp.will supply prototypes of its augmented reality headsets to the U.S. Army. But these are not your ordinary gaming models. The headsets could embrace night vision, thermal sensing and hearing protection. Ultimately the Army could cessation up purchasing up to 100,000 customized HoloLens devices, Bloomberg says.
North Sea oil blockchain platform goes live. Hopes are Vakt, created in by a consortium that includes oil majors Royal Dutch Shell, BP and Norway’s Equinor, will supplant paperwork associated with finalizing oil deals between the players, Reuters reports. The blockchain platform plans to expand to embrace crude U.S. pipelines in 2019.
Justice indict hackers in Atlanta hack. The Justice Department Wednesday indicted two Iranians for a ransomware beset earlier this year that crippled Atlanta’s government for days, the current York Times reports. The men, Faramarz Shahi Savandi and Mohammad Mehdi Shah Mansouri, are credited with creating SamSam, ransomware employed against a number of public agencies over the ultimate brace years, including the city of Newark and the Port of San Diego and a number of hospitals.
Dell resets customer passwords. Nearly a week after detecting and stopping an attempt by hackers to purloin customer data, Dell Technologies Inc. said Tuesday it would reset passwords for every of its Dell.com online store customers, Reuters reports.
Musk scraps West LA tunnel. The boring Company, Elon Musk's underground transportation venture, will cease labor on its tunnel on LA's west side after encountering resistance from community groups, Reuters said. But its labor on a towering hurry passenger tunnel between Dodger Stadium and nearby subway stops will continue.
Hello? Is it me you're looking for? As companies compete for workers in the tightest labor market in decades, more employers are rethinking the tried-and-true phone interview, rolling out one-sided, automated exchanges in which applicants give recorded responses to a chain of questions. The WSJ's Chip Cutter has more on the latest sign of the apocalypse.
Amazon store comes to NYC's SoHo. "Spending time browsing here was among my most dismal shopping experiences in recent memory," the current York Times writes. "And that was before a 20-something guy bounded into the store and started screaming: 'Alexa! Alexa! Alexa!'"
EVERYTHING ELSE YOU exigency TO KNOW
Trump administration duties absorb scrambled the global steel trade, with countries that can no longer sell into the U.S. sending their products into Europe at a record rate. (WSJ)
Renault, Nissan and Mitsubishi are “fully committed” to their global auto alliance, as top executives gathered for the first meeting of the partnership since the arrest ultimate week of its prime architect, Carlos Ghosn. (WSJ)
Deutsche Bank offices were searched by police and prosecutors seeking evidence in connection with alleged money laundering, the Frankfurt prosecutor’s office said. (WSJ)
Oil prices were hit by another wave of cumbersome selling, with fears of climbing production dragging the two most common benchmarks to lows not seen in more than a year. (WSJ)
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