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250-323 | Data Protection Administration for(R) Windows using NBU 5.0

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250-323 - Data Protection Administration for(R) Windows using NBU 5.0 - braindump

Vendor Symantec
Exam Number 250-323
Exam Name Data Protection Administration for(R) Windows using NBU 5.0
Questions 201 Q & A
Recent Update February 18, 2019
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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
Q&A : 201 Real Questions

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Symantec Data Protection Administration for(R)

Heavy pressure on Symantec because it Seeks a new CEO | killexams.com Real Questions and Pass4sure dumps

First identify: last name: email tackle: Password: confirm Password: Username:

Title: C-stage/President manager VP staff (associate/Analyst/and so forth.) Director

characteristic:

role in IT decision-making technique: Align company & IT dreams Create IT strategy check IT wants manage seller Relationships evaluate/Specify brands or companies different function Authorize Purchases now not concerned

Work cellphone: business: company dimension: trade: highway handle city: Zip/postal code State/Province: country:

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Symantec Helps find Cyber Espionage pastime focused on satellite tv for pc, Telecom, Geospatial Imaging and defense agencies in the US and Southeast Asia | killexams.com Real Questions and Pass4sure dumps

Symantec Corp. (NASDAQ: SYMC), the world’s main cyber protection company, these days introduced that Symantec’s synthetic intelligence-based mostly centered attack Analytics (TAA) expertise helped researchers expose a new assault crusade from a group called Thrip, which has infiltrated satellite tv for pc communications, telecoms, geospatial imaging, and protection agencies in the u.s. and Southeast Asia. TAA’s superior AI know-how was instrumental in the discovery of the assault, alerting Symantec’s attack Investigations crew to recreation that on the surface looked innocuous however set them on the course to uncovering the newest campaign conducted via the Thrip group. Symantec has been monitoring Thrip considering the fact that 2013, and has found out new equipment and techniques used through the group in this most fresh set of attacks.

TAA leverages AI and advanced computer researching to sweep through Symantec’s records lake of telemetry in order to spot patterns associated with centered attacks. This know-how almost automates what in the past took thousands of hours of analyst time and is obtainable in Symantec’s advanced probability insurance policy (ATP) product. From an initial alert caused via TAA in January 2018, Symantec researchers had been able to observe a path that enabled them to determine that the crusade originated from machines primarily based in mainland China. the use of these strategies, TAA detected suspicious behavior regardless of the community’s use of legit operating device points and network administration tools in an attempt to ward off detection. TAA also uncovered the use of customized malware in these attacks, as well as opting for the kinds of groups targeted. Cyber espionage is the group’s doubtless intent, however given the group has revealed a technique of compromising operational techniques, it may adopt a extra aggressive, disruptive stance may still it decide to accomplish that.

"here's likely espionage,” stated Greg Clark, Symantec CEO. "The Thrip neighborhood has been working for the reason that 2013 and their latest campaign uses ordinary operating system equipment, so focused businesses won’t observe their presence. They operate very quietly, mixing in to networks, and are most effective found out the usage of artificial intelligence that can determine and flag their actions. Alarmingly, the group appears keenly interested in telecom, satellite operators, and protection businesses. We stand ready to work with appropriate authorities to handle this serious possibility.”

Symantec has sharpened its efforts on community-resident malware, as the many vulnerabilities which are broadly standard in IOT gadgets current a new assault floor of excessive pastime.

Thrip’s attack on telecoms and satellite tv for pc operators exposes the probability that the attackers could intercept and even alter communications site visitors from enterprises and patrons. This has added to growing to be privateness concerns which have been very visible currently with the deployment of the new GDPR regulations as smartly because the VPNFilter attacks on information superhighway routers. Symantec has responded by using opening a brand new privateness center and records insurance policy lab with the intention to provide buyers with greater control over their facts, and organizations with equipment to support them responsibly manage the records they tackle. Symantec also offers a wide selection of privateness options, corresponding to Symantec VIP and Norton WiFi privacy.

Symantec has been conserving purchasers from Thrip-connected pastime since 2013. right here protections are in place to give protection to shoppers in opposition t Thrip:

File-primarily based insurance plan

  • Trojan.Rikamanu
  • Infostealer.Catchamas
  • Hacktool.Mimikatz
  • Trojan.Mycicil
  • Backdoor.Spedear
  • Trojan.Syndicasec
  • customers of Symantec’s DeepSight Intelligence Managed Adversary and hazard Intelligence (MATI) carrier have acquired distinctive experiences on "ATG14” (also referred to as Thrip), which detail strategies of detecting and thwarting activities of this adversary.

    For greater tips, please consult with the Symantec danger Intelligence blog.

    About Symantec

    Symantec organisation (NASDAQ: SYMC), the realm's leading cyber safety enterprise, helps agencies, governments and americans secure their most critical records wherever it lives. groups across the world seem to be to Symantec for strategic, integrated options to protect against refined assaults throughout endpoints, cloud and infrastructure. Likewise, a worldwide group of more than 50 million americans and households count on Symantec's Norton and LifeLock product suites to protect their digital lives at home and across their gadgets. Symantec operates probably the most world's greatest civilian cyber intelligence networks, allowing it to see and protect in opposition t essentially the most advanced threats. For more information, please consult with www.symantec.com or join with us on fb, Twitter, and LinkedIn.


    Trump blames Senate for now not approving nominees but ignores his poor record | killexams.com Real Questions and Pass4sure dumps

    Joe Davidson

    Columnist focusing on federal government concerns

    In his State of the Union address, President Trump blamed the Republican-controlled Senate for foot-dragging on his political appointees.

    “This new period of cooperation can birth with at last confirming the greater than 300 particularly certified nominees who're nevertheless stuck within the Senate — some after years of ready,” he pointed out Tuesday. “The Senate has didn't act on these nominations, which is unfair to the nominees and to our country.”

    The Republican president didn’t point out the numerous positions that stay unfilled because he hasn’t sent nominees to the Senate.

    Of 705 key political positions requiring Senate affirmation, a hundred and forty four don't have any nominee, based on an appointments’ tracker posted by The Washington publish and the Partnership for Public service, a nonpartisan respectable-government organization. About 54 % of Trump’s civilian nominees, not including judges, had been validated. That compares with President Barack Obama’s seventy seven % on the equal aspect in his presidency.

    “This administration has been the slowest at filling the jobs the first time around and the administration with probably the most turnover,” noted Max Stier, the Partnership’s president and chief govt. “The turnover is totally costly not handiest for these positions, however there also usually is a cascade have an effect on” on positions lessen within the companies.

    in lots of organizations, positions are stuffed with the aid of “appearing” individuals who do the work crucial until a confirmed respectable is in location. whereas the appearing leaders have full authority on paper, they face obstacles in follow. Stier in comparison appearing officials to change academics who might possibly be fantastic educators, but they aren’t going to strategy their roles with a protracted-time period center of attention or as massive issue solvers. “They’re no longer going to be perceived as having the same level of authority,” Stier introduced, “as someone who turned into nominated and validated by the Senate.”

    There aren't any officers, performing or otherwise, in two essential slots on the advantage systems protection Board (MSPB). It’s a small, quasi-judicial agency with a vital mandate: to shield the advantage ideas that are designed to foster a nonpartisan federal body of workers. The board hears appeals of disciplinary actions taken towards federal personnel.

    “The MSPB performs a vital function to the satisfactory of self-governance in our democratic type of govt,” noted Steve Katz, who turned into chief counsel to the MSPB chairman from 1993 to 1997. As an independent, bipartisan company, it “exists to protect the political neutrality of the federal team of workers.”

    but the three-member board has had only one member for greater than two years. The term of Mark A. Robbins, the appearing chairman, ends March 1. with no quorum the board can accomplish nothing.

    Doing nothing is verified by MSPB data. On Jan. 7, 2017, there were eleven petitions for evaluate waiting board action. through Jan. 31 this 12 months, that quantity jumped 155 times to 1,708.

    however not like many different positions in the Trump administration, there are nominees for the MSPB board. they're scheduled to be voted on with the aid of the Senate place of origin protection and Governmental Affairs Committee subsequent week. The nominees didn’t get throughout the committee final 12 months. They may make it this time, youngsters, since the Republicans have a larger majority.

    In November, all seven Democrats on the panel voted against Andrew F. Maunz, a Social protection Administration lawyer. All seven Republicans present voted for him, creating a tie. Committee Chairman Ron Johnson (R-Wis.) then decided not to carry up the other two nominees for a vote. That meant the board endured to be stranded.

    The existing circumstance “is disastrous for whistleblowers,” said Tom Devine, felony director of the nonprofit government Accountability assignment. A functioning MSPB is “their leading opportunity to attain brief relief for circumstances that commonly take over five years to make a decision. 2d, it's a loud and clear sign that Congress is not serious about the due method infrastructure to enforce the free speech it unanimously has handed 4 instances within the Whistleblower coverage Act. Third, the MSPB has the gathered experience and competencies accurately to overview preliminary Administrative choose rulings, in contrast to appellate court docket judges who may additionally most effective hear one or two govt or company whistleblower instances per yr.”

    Johnson doesn’t sound confident in regards to the vote on Wednesday.

    “i'll proceed to work with my colleagues and the administration to support be certain the advantage techniques coverage Board has the three confirmed participants essential to function properly,” he said by email. “If that is not viable, i will guide legislations that extends the time period of the ultimate remaining board member as a brief-term solution.”

    while Trump’s listing on filling govt branch political positions isn't good, he is very proud of his success in placing many judges on the federal bench. “President Trump has put in a historical variety of federal appeals court docket judges for this element of a presidency,” referred to an article posted Monday via my colleague Ann E. Marimow. On Thursday, the Senate Judiciary Committee advanced forty four judicial nominees, despite complaints by Democrats.

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    Postexposure Protection of Macaques from Vaginal SHIV Infection by Topical Integrase Inhibitors | killexams.com real questions and Pass4sure dumps

    Abstract

    Coitally delivered microbicide gels containing antiretroviral drugs are important for HIV prevention. However, to date, microbicides have contained entry or reverse transcriptase inhibitors that block early steps in virus infection and thus need to be given as a preexposure dose that interferes with sexual practices and may limit compliance. Integrase inhibitors block late steps after virus infection and therefore are more suitable for post-coital dosing. We first determined the kinetics of strand transfer in vitro and confirmed that integration begins about 6 hours after infection. We 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. We showed that gel containing the strand transfer inhibitor L-870812 protected two of three macaques when applied 30 min before SHIV challenge. We next evaluated the efficacy of 1% raltegravir gel and demonstrated its ability 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. We documented rapid vaginal absorption reflecting a short pharmacological lag time and noted that vaginal, but not plasma, virus load was substantially reduced in the breakthrough infection after raltegravir gel treatment. We 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 need to develop efficacious biomedical interventions to limit 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 have 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 be optimally formulated with a single drug or in combination for both vaginal and rectal application (4–6, 10).

    The CAPRISA 004 trial 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 dependent on adherence and was found to increase to 54% in women who reported frequent gel use (>80% adherence) compared to only 28% in women who were <50% adherent. A subsequent study (VOICE) evaluating the same 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 lack of efficacy (3). These data and similar findings from oral PrEP studies highlight the challenges of identifying intervention strategies that have both high biological efficacy and enhanced user compliance (4).

    Coitally used topical gels are important HIV prevention methods. However, to date, all such products rely on a preexposure dose that can interfere with sexual practices, and may be limited by partner’s acceptance and the need to anticipate sex. We reasoned that a topical product that can be applied by women after sex will have less impact on sexual practices, have better user control, and circumvent the need for sex anticipation and partner acceptance. These advantages can all potentially enhance compliance and effectiveness. However, all topical ARV-based gels in development use entry or reverse transcriptase (RT) inhibitors that block 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) have provided important 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 need to adequately dose mucosal tissues with RT inhibitors before virus exposure and suggest that drugs from different classes that target later steps in virus infection may be more efficacious for PEP.

    HIV or SIV integration into the host DNA is the last step before an irreversible infection of a cell takes place. Integration is a multistep process catalyzed by the viral integrase that follows reverse 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 be inhibited at various stages by three main classes of integrase inhibitors: 3′-processing inhibitors (13), strand transfer inhibitors (14), and macromolecular complex LEDGF/p75-IN inhibitors (15). Strand transfer inhibitors are the most developed and have 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 ability to chelate the Mg2+ ions in the catalytic core domain of integrase, thereby disrupting the interaction between the integrase/viral complementary DNA preintegration complex 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, all 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 be effective in reducing SIV replication in macaques (16). For PEP, blocking the last step in virus integration by strand transfer inhibitors can be 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 range (1 to 4 nM), which provides an opportunity to test in macaque models the concept of topical prophylaxis by this class of drugs for both pre- and postexposure protection (16, 17).

    Here, we used a repeat 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 use of pigtailed macaques, which have a menstrual cycle and vaginal anatomy similar to women, an inoculum dose with viral RNA levels in the range 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 have the added advantage of increasing statistical power without requiring large groups of animals because they allow measurement of protection over more than one transmission event per animal. We also 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 active for up to 1 year at room temperature. To investigate whether biological factors present in the vagina, including mucus, enzymes, and microflora, may compromise the stability and potency of RAL gel, we assessed the antiviral activity of RAL gel incubated in cervicovaginal fluids (CVFs). Figure 1 shows that CVF had no impact on RAL activity because no difference 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 reverse transcription and integration and to inform the dosing modality in vivo, we 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. Figure 2 shows that TFV maintained high (>95%) protection up to 2 hours after infection, but was only ~50% protective when added at 5 hours after infection. In contrast, RAL provided high protection (>90%) at 6 hours and remained above 50% protection for up to 10 hours after infection. These data suggest that strand transfer is delayed by at least 4 hours after reverse transcription, providing a longer window to block infection with integrase inhibitors than inhibitors of reverse 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). All controls received the same placebo gel and were challenged with the same virus stock under identical experimental conditions. In this model, pigtailed macaques are more susceptible to infection during the late luteal phase of the menstrual cycle (10, 23). As previously described, the risk for infection was calculated per month or menstrual cycle (10). Figure 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 function 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 include 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. Figure 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. All 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. All 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 suggest that topically applied integrase inhibitors can prevent 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 also important for determining impact on acute viremia and emergence of drug resistance in animals that fail gel prophylaxis and continue receiving gel treatment. We measured L-812 and RAL concentrations in plasma 30 min after each gel administration to determine systemic drug exposure at time of challenge and also assessed the impact of the menstrual cycle on vaginal drug absorption. We have previously noted 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 phase when the vaginal epithelium is thin (24, 25). We analyzed longitudinally both progesterone and drug concentrations in plasma in all macaques that were treated with L-812 or RAL gel (Fig. 4, A and B). With the exception of one (PMl), all macaques experienced at least two menstrual cycles during the study period. Plasma drug concentrations 30 min after vaginal dosing showed similar trends in all animals with substantially higher concentrations consistently detected after peak progesterone. The mean 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 confirm 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 do not show a clear 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 show progesterone concentrations. (B) Six macaques dosed twice weekly with RAL gel (dotted line); shaded areas show progesterone concentrations. All 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. Figure 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 difference 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; large circles) or placebo (n = 4, black dotted lines). Filled symbols indicate samples sequenced for drug resistance in plasma and vaginal fluids. The broken line denotes the limit 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 important 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 all revealed wild-type genotypes (Fig. 5A). To examine drug resistance emergence at the site of gel application, we also genotyped SHIV RNA isolated from vaginal secretions in the RAL breakthrough infection (fig. S1). Vaginal fluids with detectable SHIV (n = 4) were genotyped, and all found to be wild type (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, our data support exploring new post-coital modalities with integrase inhibitors for enhanced acceptance and compliance in women.

    We used single-round infections to determine the kinetics of reverse transcription and integration and show 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 take longer in less activated primary cells that are commonly present in vivo in which reverse transcription and integration proceed more slowly than in transformed cell lines (26, 28). Thus, it is likely that the dosing window in vivo may be 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 decay to suboptimal levels (19). We also 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 lack of intracellular activation suggest that the pharmacological lag time for in vivo activity may be 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 also document in pigtailed macaques changes in plasma drug levels during the menstrual cycle and observe higher concentrations during the progesterone-dominated luteal phase. These results reflect a higher vaginal permeability peaking during the luteal phase 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 also explains the cyclical changes in plasma concentrations of TFV and FTC we have previously observed in pigtailed macaques dosed with gels containing TFV and FTC (6). We also examined the relationship between plasma drug concentrations and vaginal efficacy because Cranage et al. previously found a positive association between TFV concentration in plasma and the degree of rectal protection by rectal TFV gel (9). With only two breakthrough infections, we were unable to discern any trend because one animal had high 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 show the usefulness of the pigtailed macaque model that has a menstrual cycle similar to that of women, and support 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, we found 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 have 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 large surface area of the vagina for a significant period of time (6, 7, 10). Previous work with TFV gels in macaques has shown that the pharmacologically active TFV-diphosphate (TFV-DP) concentrations in vaginal lymphocytes predict 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 also apply to RAL. However, confirming this correlate will require measuring intracellular RAL concentrations in vaginal lymphocytes, which could be 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 also used for treatment. Prolonged systemic treatment of SHIV-infected macaques with L-812 has been found to select drug-resistant variants with the integrase mutation (N155H) (20). We show 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, we found 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 differ from those in macaques that fail oral PrEP in which reductions in virus loads and selection of drug-resistant variants have been observed (7, 9). Given that the antiviral activity from gels remains largely concentrated in the vaginal compartment, we also evaluated the viral population in the genital secretions from the RAL breakthrough infection. We detected only wild-type SHIV, but noted 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 also have reduced drug resistance selection. These data are only from one animal and require confirmation, but they do 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 include 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 suggest little 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, all of which can increase 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, we show preclinical in vivo data that support the use of topical integrase inhibitors for HIV prevention. We highlight the advantage of using this class of drugs in new 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 small molecules that do 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 also 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 room 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 all 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 all 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). All 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. All infected macaques continued to receive study gel (placebo, L-812, or RAL) twice weekly for up to 8 weeks after infection. All experiments were done under highly controlled conditions by the same personnel with the same virus stock, inoculum dose, and procedures as described in previous studies (6). These studies adhered to the Guide for the Care and Use of Laboratory Animals (Institute for Laboratory Animal Research, 1996); all procedures were approved by the Institutional Animal Care and Use 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 reverse 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 standard sequence analysis of SIVmac239 integrase reference sequence base pairs 3624 to 4332 (amino acids 1 to 234). CVF specimens were collected by instilling and recollecting about 5 ml of sterile 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 standard (RAL was used as an internal standard 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 method 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 run 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 limit of quantification of 5 ng/ml for both analytes and standard 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 Center 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 phase 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 proportion 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 all 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.

  • Acknowledgments: We thank S. Ehnert, C. Souder, E. Strobert, and the animal care staff at the Yerkes National Primate Center (Emory University) as well as J. Mitchell, E. Sweeney, and S. Bachman at the CDC for monitoring, maintaining, and performing animal procedures using our macaque cohort. The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the CDC. Use of trade names is for identification purposes only and does not constitute endorsement by the U.S. CDC or the Department of Health and Human Services. Funding: Partially supported by Interagency Agreement Y1-AI-0681-02 between CDC and NIH. Author contributions: C.D., F.N., J.S., D.H., J.G.G.-L., and W.H. designed the research. C.D. and S.S. executed the macaque studies. C.D., S.S., U.M.P., R.W., and A.T. performed in vitro experiments and provided assistance in processing macaque samples and data analysis. J.L. performed drug resistance testing. A.M. and C.-P.P. performed drug analysis. D.L.H. performed statistical analyses. D.H. provided drug. C.D. and W.H. wrote the manuscript. Competing interests: W.H. and J.G.G.-L. are named on patent application 11/669,547 filed by the U.S. government entitled “Inhibition of HIV infection through chemoprophylaxis.” The authors declare that they have no competing interests.

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    Implementing Proxy Server | killexams.com real questions and Pass4sure dumps

    Designing a Proxy Server Implementation

    Before you can design a Proxy Server implementation and install Proxy Server, you need to be knowledgeable on a number of concepts:

  • IP routing concepts

  • Firewalls concepts

  • Packet filtering concepts

  • Files and protocols utilized in Web applications

  • To design a Proxy Server implementation, there are a number of factors that has an impact on the Proxy Server design:

  • The characteristics of data that will pass to the Proxy Server. Data characteristics should include factors such as the quantity of data which you expect the Proxy Server to handle, and whether data confidentiality needs to be ensured.

  • The type of firewall that the Proxy Server will interface with.

  • Decide whether the Proxy Server will be located within the DMZ or on the edge of the network.

  • Determine the correct sizing of Proxy Server(s).

  • The resources located on the private network which Internet users should be able to access.

  • The physical layout of the servers

  • The connections to and from proxy servers.

  • The volume of expected network traffic.

  • Bandwidth between sites.

  • Who needs access to the Proxy Server, and what type of access is required.Implementing Proxy Server

  • Determine the time which users should be able to access the Proxy Server.

  • Future network expansion.

  • Existing proxy server configuration: Here, factors such as the location of an existing proxy server, the WAN connections being used, and the protocols used in the private network should be considered.

  • After you have looked at all the factors which impact the Proxy Server design, you have to determine, or select between a few additional design elements:

  • The type of connection that the Proxy Server must support:

  • The type of services that the Proxy Server must provide:

  • Web Proxy

  • WinSock Proxy

  • Socks Proxy

  • Packet filtering

  • Reverse Web Proxy

  • The types of connection technology that the Proxy Server must support:

  • Digital Subscriber Line (DSL)

  • Integrated Services Digital Network (ISDN)

  • Public Switched Telephone Network (PSTN)

  • T1

  • X.25

  • The type of Proxy Server clients that the Proxy Server must support.

  • The type of routing which each router should support:

  • Dynamic routing

  • Static routing

  • Whether multiple proxy servers will be implemented to improve performance and provide high levels of availability. Proxy Server provides a feature called proxy arrays. A proxy array is a solution whereby one or multiple proxy servers operate as a single cache for client requests. Benefits provided by the proxy array feature include scalable performance, and fault tolerance.

  • You can create a Proxy Server design where frequently requested content is cached. Proxy Server can locally cache Internet sites and files which are frequently requested. Subsequent requests for these Internet sites are then serviced from the local cache. Cached information is accessed by users from a location on the Local Area Network (LAN). This design has a number of benefits. For instance, bandwidth utilization to the Internet ends up being reduced because cached information does not need to be downloaded from the Internet. All of this leads to an improvement in the service experienced by users.

    With passive caching, Proxy Server stores objects in the Proxy Server cache with each object obtaining a Time To Live (TTL) value. Before Proxy Server forwards requests to the Internet, it first checks the Proxy Server cache to determine if the request can be serviced from there. Active caching works together with passive caching. With active caching, Prox Server automatically generates requests for specific objects in the Proxy Server cache so that frequently requested objects remain cached.

    The requirements for creating a Proxy Server design that caches content are listed here:

  • Web content is only cached on NTFS partitions. This basically means that you need to have, minimally, one NTFS partition that is capable of storing frequently accessed Web content.

  • You should have two network adapters so that private network traffic with Internet traffic can be separated. This results in network congestion being reduced.

  • You should place proxy servers using their purpose or function as the basis to determine placement. This concept is illustrated here:

  • If the Proxy Server is to provide connectivity between the private network and the Internet;

  • If the Proxy Server is to cache Web content so that frequently accessed content can be accessed from the local cache;

  • If the Proxy Server is to is to provide both connectivity between the private network and the Internet and cache Web content;

  • The following information has to be defined for every interface in the Proxy Server implementation:

  • The type of connection (persistent/nonpersistent) between the router interface and the network.

  • The following IP information for interfaces that are connected to IP network segments:

  • IP address configuration.

  • IP subnet mask configuration.

  • The following IPX information for interfaces that are connected to IPX network segments:

  • IPX network number.

  • IPX frame type

  • Another component that should be included when you plan your Proxy Server implementation is to determine the client operating systems that Proxy Server should support. Proxy Server can support a number of different client operating systems.

    You should define Proxy Server client support based on what your Proxy Server implementation should provide:

  • Define Windows Proxy Server client support for the following reasons:

  • All Windows operating systems should be supported.

  • IP traffic needs to be redirected through Proxy Server Support IPX to IP gateways.

  • Clients should utilize the local address table (LAT) to determine the destination IP addresses.

  • Define default gateway support for the following reasons:

  • Define Microsoft Internet Explorer 5.0 support for the following reasons:

  • All operating systems that include Internet Explorer 5.0 should be supported.

  • Only HTTP and FTP traffic will pass through Proxy Server.

  • The Internet Explorer Administrator Kit (IEAK) is to be used to administer Proxy Server client configuration.

  • Define SOCKS support for the following reasons:

  • Support for Unix and Macintosh is ensured.

  • All operating systems that utilize SOCKS standard should be supported

  • IPs supported by SOCKS applications should be redirected.

  • Another planning component that should be included when you design your Proxy Server implementation is to determine the level of data protection that should be configured.

  • Inbound and outbound packet filters can be configured to filter and restrict traffic, based on the criteria defined for the different IP traffic types.

  • Domin filters can be configured to restrict Internet access to only certain IP addresses or FQDNs. In a domain filter, you can include a number of Internet sites and then define the action that the domain filter should take when a request is received for one of these sites: Reject packets for these specific Internet sites and forward all other packets OR forward packets to these specific Internet sites and reject all other packets. Domain filters can restrict outbound traffic, based on a single computer or the IP address of a cluster, an IP address range or a FQDN.

  • You can utilize Proxy Server user authentication to specify Internet access, based on user or group account.

  • Through Web publishing, you can restrict inbound traffic based on the URL requests of Internet users.

  • The default configuration of Proxy Server is to drop the URL requests of Internet users. This means that Internet users do not have access to Web and FTP servers hosted within the private network, by default. You can though define URLs where requests for these URLs should be passed to Web and FTP servers on the private network. Proxy Server will allow URL requests when you define them in the Web Publishing list.

    For URLs that are requested which are defined in the Web Publishing list, Proxy Server passes the requests to the Web and FTP servers on the private network.

    For URLs that are requested which are not defined in the Web Publishing list, Proxy Server performs either of the following:

    There are also a number of techniques that optimize Proxy Server performance, which you should consider implementing:

  • Caching Web content improves performance. Cached information is accessed by users from a location on the Local Area Network (LAN). This means that bandwidth utilization to the Internet ends up being lowered because cached information does not need to be downloaded from the Internet. All of this leads to an improvement in the service experienced by users.

  • Proxy Server also provides a feature called proxy arrays. A proxy array is a solution whereby one or multiple proxy servers operate as a single cache for client requests. Benefits provided by the proxy array feature include scalable performance, and fault tolerance.

  • Network Load Balancing (NLB) can be used to distribute the processing load of inbound traffic over multiple proxy servers. This leads to high availability and performance optimization.

  • Round Robin DNS can also be used to load balance inbound traffic across multiple proxy servers, thereby also providing high availability and performance optimization.

  • The advantages of using proxy arrays as a Proxy Server optimization method when you implement Proxy Server are listed here:

  • Because Web content is cached over multiple servers, no single server hosts all Web content.

  • If a server in the proxy array fails, failover is immediately provided.

  • The advantages of using Network Load Balancing (NLB) as a Proxy Server optimization method when you implement Proxy Server are listed here:

  • You can add or remove proxy servers residing in the NLB cluster.

  • Load balancing occurs dynamically over all proxy servers residing in the NLB cluster.

  • Because load balancing and the addition or removal of proxy servers occurs dynamically, availability and performance is improved.

  • The NLB cluster is automatically reconfigured when a proxy server happens to fail.

  • The advantages of using Round Robin DNS as a Proxy Server optimization method when you implement Proxy Server are listed here:

  • Load balancing is performed on all proxy servers in the round robin DNS.

  • Round Robin DNS can operate on all operating system platforms.

  • Performance is improved because traffic is basically load balanced over all proxy servers.

  • If you need to provide the highest possible level of server availability for your Proxy Server implementation, you should use Microsoft Windows Clustering. Using Microsoft Windows Clustering provides the following benefits for your Proxy Server implementation:

  • The Proxy Servers all share a common cache.

  • If a server in the proxy array fails, failover is immediately provided.

  • Because the cache does not need to be built again when a server fails, restore occurs quite faster.

  • To optimize Internet access, you can include the following Proxy Server caching methods in your Proxy Server design:

  • As mentioned previously, with passive caching, Proxy Server stores objects in the Proxy Server cache with each object obtaining a Time To Live (TTL) value. Before Proxy Server forwards requests to the Internet, it first checks the Proxy Server cache to determine if the request can be serviced from there. When the Proxy Server cache becomes full, Proxy Server removes objects from the cache, based on a combination of factors: object size, object age, and object popularityThe advantages of using passive caching in your Proxy Server implementation are:

  • With Active Caching, Proxy Server automatically generates requests for specific objects in the Proxy Server cache so that frequently requested objects remain cached. Proxy Server determines which objects should be flagged for active caching by considering object popularity, Time To Live (TTL) value of objects, and server load to determine the level of active caching performed.The advantages of using active caching in your Proxy Server implementation are:

  • Determining Proxy Server Hardware and Software Requirements

    Proxy Server has a few minimum hardware and software implementation requirements. However, depending on the size of the organization, existing hardware and software, future network expansion, and expected traffic volumes; the Proxy Server implementation requirements between organizations would differ. For each different network environment, there are different requirements for a Proxy Server implementation.

    The requirements listed below merely serves as a guideline on the hardware requirements for a Proxy Server implementation:

  • Processor; Intel 486 or faster supported RISC-based microprocessor

  • Disk space; 10 MB available disk space for Proxy Server

  • For caching; 100 MB plus an additional 0.5 MB for each Web Proxy service client.

  • RAM; at least 24 MB. For RISC-based systems, this increases to 32 MB.

  • An NTFS formatted partition to store the Proxy Server cache.

  • A network adapter card for connection to the LAN.

  • A network interface configured for the Internet.

  • When planning a Proxy Server implementation, you have to decide on the hardware that you will used to establish connections to the Internet:

  • ISDN lines can be used to establish connections to the Internet. ISDN is a digital dial-up service that utilizes telephone cabling and other technology to provide Internet connections. The different types of ISDN services are ISDN Basic Rate Interface (BRI) and ISDN Primary Rate Interface (PRI).The main characteristics of ISDN Basic Rate Interface (BRI) are listed here:

  • BRI connections work well for small companies

  • BRI connections are available from quite a number of telephone companies.

  • ISDN BRI can offer 128 Kbps of bandwidth.

  • Provide e-mail for a maxmum of 20 concurrent users.

  • Provide large FTP downloads for only 3 to 4 simultaneous users.

  • Provide Web browsing for 6 to 8 concurrent users.

  • The main characteristics of ISDN Primary Rate Interface (PRI) are listed here:

  • ISDN PRI can offer 1.544 Mbps transmission speed.

  • Provide e-mail for a maximum of 120 concurrent users.

  • Provide large FTP downloads for only 40 to 50 simultaneous users.

  • Dial-up modem connections are ideal if your organization only consists of a small number of users that do not need to connect to the Internet on a regular basis. This is due to dialup modem connection only being able to meet the bandwidth requirements of a small number of users. Modems can be installed on a computer, and then shared through the Windows Internet Connection Sharing (ICS) service.A few characteristics of dial-up modem connections are:

  • A dial-up modem connection can only reach up to 53 Kbps.

  • Provide e-mail for a maximum of 10 concurrent users.

  • Provide large FTP downloads for only 1 to 2 simultaneous users.

  • Provide Web browsing for 2 to 3 concurrent users.

  • You also have to decide on the hardware which will be utilized to connect the server to the Internet:

  • Analog modem: Analog modem run at 28.8 or 33.6 Kbps speeds. An analog modem is ideal for a single user connecting to the Internet, and for a networked server gateway.

  • ISDN adapters: This is the popular choice. The ISDN adapters dial an ISDN access number and then maintain the particular connection.

  • Routers: Routers are networking devices that connect networks.

  • Installing Proxy Server

    You should verify a number of things before you actually install Proxy Server:

  • 10 MB available disk space for Proxy Server and 100 MB plus an additional 0.5 MB for each Web Proxy service client.

  • An NTFS formatted partition to store the Proxy Server cache.

  • TCP/IP should be installed on the computer.

  • The internal network interface should be bound to the TCP/IP or IPX/SPX protocol being used on the LAN.

  • You should configure the software for two network adapter cards before you attempt to install Proxy Server.

  • When Proxy Server is installed, the following changes are made to the computer on which you are installing it:

  • The Web Proxy service is installed.

  • The WinSock Proxy service is installed.

  • The Socks Proxy service is installed.

  • Each of these services is added to the Internet Service Manager administration tool.

  • The local address table is installed.

  • On the NTFS volume, the cache drive is created.

  • The client installation and configuration software is copied.

  • The Mspclnt shared folder is created.

  • The Proxy Server Performance Monitor counters are installed.

  • The HTML online documentation is installed.

  • How to install Proxy Server
  • On the Proxy Server installation CD, proceed to run Setup.

  • Click Continue on the Welcome to the Microsoft Proxy Server Installation program screen.

  • The Microsoft Proxy Server Setup page opens.

  • Specify the 10-digit product key provided on the CD-ROM case. Click OK.

  • The Microsoft Proxy Server Setup dialog box displays the default destination folder and the Installation Options button. Click the Installation Options button.

  • The Microsoft Proxy Server – Installation Options dialog box opens, displaying all components as being selected. Click Continue.

  • Setup now stops the Web services.

  • The Microsoft Proxy Server Cache Drives dialog box opens. Caching is by default enabled.

  • The local drives of the server are listed in the Drive box.

  • Select the drive which should be used to store cached data. In the Maximum Size (MB) box, enter th appropriate value. Click Set, and then click OK.

  • The Local Address Table Configuration dialog box opens.

  • Click the Construct Table button.

  • The Construct Local Address Table dialog box opens.

  • Select Load from NT internal Routing Table to select the network adapter cards thats IP addresses must be added to the local address table.

  • Select the Load known address ranges from the following IP interface cards option, and then select the network adapter. Click OK.

  • Click OK to acknowledge the message displayed, indicating that the IP addresses have been loaded into the local address table.

  • The Local Address Table Configuration dialog box opens, displaying IP addresses in the Internal IP Ranges box.

  • Check that the addresses defined are correct, and then click OK.

  • The Client Installation/Configuration dialog box opens.

  • Enter the appropriate information and verify that the correct computer name is displayed in the Computer name field and Proxy field.

  • If you enable the Automatically configure Web browser during client setup checkbox, the Web browser network configuration setting of the client is changed so that client requests are sent to the Proxy Server, and not to the Internet.

  • Click Configure.

  • You can either run the default script to configure the client Web browser, or alternatively, you can use a custom URL.

  • Click Properties located beneath Browser automatic configuration script.

  • The Advanced Client Configuration dialog box opens.

  • Specify whether the Proxy Server is utilized for local servers.

  • Specify the IP addresses that should be excluded from Proxy Server.

  • Specify the domains that should be excluded from Proxy Server.

  • Specify a backup to the proxy server.

  • Click OK.

  • The Access Control dialog box opens.

  • Verify that access control is specified for the Web Proxy service and for the WinSock Proxy service and then click OK.

  • The Proxy Server Setup files are copied to the computer.

  • When the Setup Information dialog box opens, click OK. The Setup Information dialog box displays information on the packet filtering feature. The packet filtering feature is not automatically enabled when Proxy Server is installed. Click OK.

  • A Proxy Server 2.0 Setup was completed successfully message is displayed.

  • How to install WinSock Proxy Client on client computers

    When you install WinSock Proxy Client on client computers, the following changes are made:

  • The Proxy Client program group is created

  • The local address table file, Msplat.txt, is installed on the client. Proxy Server will update this file.

  • Mspclnt.ini is also copied to the client.

  • The WSP Client icon is added to Control Panel. This only occurs for Windows 3.x, Windows 95, and Windows NT clients.

  • Remote WinSock from WinSock Proxy Client replaces Winsock.dll. This would enable the computer to only access Internet sites using the WinSock Proxy service.

  • To install WinSock Proxy Client on a client computer;

  • Open Internet Explorer

  • In the Address box, enter http://proxycomputername/msproxy.

  • The WinSock Proxy Client 2.0 Installation page is displayed.

  • To install WinSock Proxy Client, click WinSock Proxy 2.0 client.

  • Click the Open it option and click OK.

  • The Microsoft Proxy Client Setup dialog box opens.

  • Click Continue to proceed with the installation.

  • Click Install Microsoft Proxy Client to start copying Setup files to the client computer.

  • Click OK.

  • The Setup – Restart System dialog box opens.

  • Click the Restart Windows Now option.

  • How to add or remove Proxy Server components
  • On the Proxy Server installation CD, proceed to run Setup.

  • Click Add/Remove on the Setup screen.

  • Follow the instruction displayed to add or remove Proxy Server components.

  • How to restore Proxy Server settings or files
  • On the Proxy Server installation CD, proceed to run Setup.

  • Click Reinstall on the Setup screen.

  • Follow the instructions displayed to restore Proxy Server settings/files.

  • How to remove Proxy Server from the server
  • On the Proxy Server installation CD, proceed to run Setup.

  • Click Remove All on the Setup screen.

  • Click Yes to acknowledge that you want to remove Proxy Server.

  • Proxy Server is then removed from the server.

  • How to disable WinSock Proxy Client
  • Open Control Panel.

  • Double-click WSP Client.

  • Deselect the Enable WinSock Proxy Client checkbox.

  • Restart the computer.

  • How to re-enable the WinSock Proxy Client
  • Open control Panel.

  • Double-click WSP Client.

  • Check the Enable WinSock Proxy Client checkbox.

  • Restart the computer.

  • Administering Proxy Server using the Internet Service Manager

    You can use the Internet Service Manager to configure properties for the Web Proxy, WinSock Proxy, and Socks Proxy services of Proxy Server.

    To open the Internet Service Manager;

  • Click Start, click Programs, click Microsoft Proxy Server, and then click Internet Service Manager.

  • You can open the properties of specific Proxy Server service by double-clicking the computer name displayed alongside the particular service name.

  • There are some properties settings which are common for all three Proxy Server services, and there are others that are relevant for only a particular Proxy Server service. This concept is illustrated here.

  • The properties settings which can be configured for the Web Proxy service are listed here:

  • Service

  • Permissions

  • Caching

  • Routing

  • Publishing

  • Logging

  • The properties settings which can be configured for the WinSock Proxy service are listed here:

  • Service

  • Permissions

  • Protocol

  • Logging

  • The properties settings which can be configured for the Socks Proxy service are listed here:

  • Service

  • Permissions

  • Logging

  • The configuration settings which you can view and configure on the Service tab for each of the three Proxy Server services are listed below:

  • View the product release.

  • Verify the product ID.

  • Add additional information on the service.

  • Add additional information on the server.

  • View the current sessions.

  • Navigate to the Shared services tabs.

  • Navigate to the Configuration tabs.

  • The configuration settings which you can view and configure on the Permissions tab for the Web Proxy and WinSock Proxy services are listed below:

  • Select or disable the Enable access control checkbox.

  • Select the Protocol when defining user or group permissions. Permissions are basically assigned for each protocol.

  • Define user and group permissions for using the Internet protocols.

  • The configuration settings which you can view and configure on the Permissions tab for the Socks Proxy are:

  • Specify the source and destination for an entry, and then define whether requests should be allowed or defined.

    The configuration settings which you can view and configure on the Caching tab for the Web Proxy and service is listed here:

  • Select the Enable caching checkbox and then select between the following Cache expiration policy options:

  • Select the Enable active caching checkbox and then select between the following options:

    Faster user response is more important.

  • The configuration settings which you can view and configure on the Routing tab for the Web Proxy and service is listed here:

  • For upstream routing, you can select between the following options:

  • Use direct connection.

  • Use Web proxy or array.

  • If you select the Enable backup route checkbox, you can select between the following options:

  • Use direct connection.

  • Use Web proxy or array.

  • You can also select the Resolve Web proxy requests within array before routing upstream checkbox on this tab.

  • The configuration settings which you can view and configure on the Publishing tab for the Web Proxy and service is listed here:

  • Enable/disable Web publishing.

  • Configure computers to publish information on the Internet via the Proxy Server.

  • Specify what should happen to incoming Web server requests:

  • The configuration settings which you can view and configure on the Logging tab for all three Proxy Server services are listed below:

  • Enable/disable logging. When enabled, the following types of information will be logged:

  • Server

  • Client

  • Connection

  • Object

  • Specify the Log to file option, or the Log to SQL/ODBC database option.

  • Specify when a new log should be opened.

  • Specify the log file directory.

  • How to disable IP routing (control access to the private network)
  • Open Control Panel

  • Double-click Network.

  • The Network dialog box opens.

  • Click the Protocols tab.

  • Select TCP/IP, and click Properties.

  • The TCP/IP Properties dialog box opens.

  • Switch to the Routing tab.

  • Ensure that the Enable IP Forwarding checkbox is not selected (blank).

  • Click OK.

  • How to configure publishing configuration settings for the Web Proxy service
  • Open Internet Service Manager.

  • Double-click the computer name alongside the Web Proxy service.

  • The Web Proxy Service Properties dialog box opens.

  • Click the Publishing tab.

  • Select the Enable Web publishing checkbox.

  • If you want to drop all incoming Web server requests, click the Discard option.

  • If you want to forward all incoming Web server requests to IIS on the Proxy Server computer, click the Sent to the local Web server option.

  • If you want forward all incoming Web server requests to a specific downstream server, click the Sent to another Web server option.

  • If you want to configure the default Web server host, click Default Mapping.

  • The Default Local Host Name dialog box opens.

  • Provide the name of the default server. Click OK.

  • Click Apply and click OK.

  • How to enable dynamic packet filtering
  • Open Internet Service Manager.

  • Double-click the computer name alongside the Web Proxy service.

  • The Web Proxy Service Properties dialog box opens.

  • Click the Security button on the Service tab.

  • Click the Packet Filters tab.

  • On the Packet Filters tab, click the Enable packet filtering on external interface checkbox.

  • Select the Enable dynamic packet filtering of Microsoft Proxy Server packets checkbox.

  • Click OK.

  • Click OK in the Web Proxy Service Properties dialog box.

  • How to create a packet filter using predefined protocol definitions
  • Open Internet Service Manager.

  • Double-click the computer name alongside the Web Proxy service.

  • The Web Proxy Service Properties dialog box opens.

  • Click the Securty button on the Service tab.

  • Click the Packet Filters tab.

  • On the Packet Filters tab, click Add.

  • When the Packet Filter Properties dialog box opens, click the Predefined filter option.

  • Select a protocol from the available Protocol ID list.

  • In the Local host area of the Packet Filter Properties dialog box, select the appropriate option to allow packet exchange with a host.

  • In the Remote host area of the Packet Filter Properties dialog box, specify one host or the Any host option.

  • Click OK.

  • How to create a packet filter using custom protocol definitions
  • Open Internet Service Manager.

  • Double-click the computer name alongside the Web Proxy service.

  • The Web Proxy Service Properties dialog box opens.

  • Click the Security button on the Services tab.

  • Click the Packet Filters tab.

  • On the Packet Filters tab, click Add.

  • When the Packet Filter Properties dialog box opens, click the Custom filter option.

  • Select a protocol from the available Protocol ID list.

  • Select a direction from the Direction list.

  • Select an option from the available options in the Local port area.

  • Select either the Any option or Fixed port option in the Remote port area.

  • In the Local host area of the Packet Filter Properties dialog box, select the appropriate option to allow packet exchange with a host.

  • In the Remote host area of the Packet Filter Properties dialog box, specify one host or the Any host option.

  • Click OK.

  • How to change the packet filter list entries
  • Open Internet Service Manager.

  • Double-click the computer name alongside the Web Proxy service.

  • The Web Proxy Service Properties dialog box opens.

  • Click the Security button on the Service tab.

  • On the Packet Filters tab, click the Enable packet filtering on external interface checkbox.

  • Select the Enable dynamic packet filtering of Microsoft Proxy Server packets checkbox to enable dynamic packet filtering.

  • Click the Edit button.

  • The Packet Filter Properties dialog box opens. Change the necessary settings and then click OK.

  • If you want to remove a filter, click the Remove button.

  • Click OK.

  • How to configure Proxy Server logging
  • Open Internet Service Manager.

  • Double-click the computer name alongside the Web Proxy service.

  • The Web Proxy Service Properties dialog box opens.

  • Click the Security button on the Service tab.

  • Click the Logging tab.

  • Click the Enable logging using checkbox.

  • Select the appropriate format in the Format list box.

  • Click OK.

  • How to back up a Proxy Server configuration
  • Open Internet Service Manager.

  • On the View menu item, click Servers View.

  • Double-click the computer name, and then double-click Web Proxy (Running).

  • The Web Proxy Service Properties opens.

  • Click the Service tab.

  • In the Configuration area, click Server Backup.

  • When the Backup dialog box opens, verify the information shown on where the backup file will be saved

  • Click OK to create a back up of the Proxy Server configuration.

  • How to restore a Proxy Server configuration
  • Open Internet Service Manager.

  • Double-click the computer name, and then double-click Web Proxy service.

  • The Web Proxy Service Properties opens.

  • Click the Service tab.

  • In the Configuration area, click Server Restore.

  • When the Restore Configuration dialog box opens, click the Browse button to select the Proxy Server configuration file.

  • Select the Proxy Server configuration file that you want to use for the restore.

  • Click Open.

  • Select the Full Restore option.

  • When the Restore Configuration dialog box opes, click OK to start the restore of the Proxy Server configuration.



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