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The national Q1 results of multiple sources are flooding in, and the prognosis looks decidedly positive for labor in the United States for the coming months.
Recent research data suggests that hiring among the private sector was slightly up, compared to what was expected. News of the increase, in the manner of more than 200,000 jobs, bodes well for the US labor market as WSJ analysts also predict 190,000 new jobs. According to the report, medium-sized businesses were among the biggest winners, as they hired 82,000 in January 2016, a definite uptick from December's 77,000. Further, with unemployment rates of 5 per cent holding at their lowest levels in 7 ½ years (according to Reuters) the job market is looking strong for America.
As a business grows from strength to strength, becoming more successful and moving from a small business to a medium sized business with hundreds of employees and beyond, strategic and timely investments, particularly in IT and HR infrastructure and technology, have the power to yield even greater gains in terms of profitability.
Rather than resisting technology due to fear or lack of knowledge, it is important to start thinking about how you can embrace it and use it to enhance your business.

subset: commoncrawl


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*Still interested in the bundle but want to swap out a design? Email us at collegemagazineshop.com.
We Also Recommend


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5 Lessons Learned: Homes
May 31, 2018
Homes for Sale
If one has dreamed of buying his or her own home for many years, one might be feeling so excited today, as he or she plans to find that dream home. One might also feel a little bit of nervousness, however, as there is no doubt that he or she knows that this financial step is one that is very large, one that does require commitment. One will be glad to know that when he or she is able to find the best house to buy, the benefits will be so much more than the challenges that he or she might face. Those who buy their own homes, then, especially if it is in a good area, can be sure that, when they do so, they will be able to enjoy all of these benefits.
Buying a house in a good area is certainly something that will be beneficial to you in a lot of ways, one of which is the fact that when you do so, you can be sure that you are making, for yourself and for the future, one of the very best investments that you can ever make today. The reason why this is true is because property is very valuable today, and what is more, its value is expected to rise even more as the years go on. Those who wish to sell the homes that they buy someday, then, can be sure tht they will be able to enjoy gains through doing so.
If one decides to buy a house in a good area, he or she will also be glad to know that, through owning such a house, security can be enjoyed as time goes on. One might be living in a house that is merely rented, and if this is so, he or she might know that this is a situation that is but temporary, meaning that nothing is secure or stable, and everything can change without warning. You can be sure that, on the other hand, if you buy your own home, you do not need to worry about your landlord changing his mind and forcing you to move, and this will give you security.
Buying your own house in a good area is also something that you should do, as when you do so, you can be sure that it will give you great satisfaction. You might just imagine the feeling of owning your own home, one that is beautiful and which fits your own tastes perfectly – you might just feel how wonderful this will be, and what great pride you can feel in it.
Buying a house in a good area, then, is certainly something that you should do, as when you do so, you can be sure that the benefits and advantages which are sure to come your way will be wonderful and worthwhile altogether.


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There is a situation that has had little press but is a bit disturbing….well to me at least…..
Pres. Trump is snuggling up to those authoritarian leaders on the planet…..and that is in line with previous admins…..the nation has snuggled up with the likes of Saddam, Mubarak, the Saudis, Noriega, etc…..so this is something in-line with our policies in the past. But is it the best policy?
Trump took office in mid-January and soon met with the King Of Jordan…..an authoritarian figure in his own right……but a solid ally to the US…
the Trump administration — which has defined its top priority as defeating the Islamic State — needs stability in Jordan and a close strategic relationship with Amman. The kingdom is a major base of U.S. air operations for the campaign in Syria, a key intelligence sharing partner, and a leading Arab ally in the battle of ideas against radical Islam. Recognizing this importance, the United States provided nearly $1.6 billion in military and economic assistance to Jordan last year — a figure that represented more than 10 percent of the kingdom's annual budget and made it the second-largest recipient of U.S. foreign assistance dollars in the world.
Then along came the strongman of Egypt, al-Sisi……slinking into DC…….since coming to power Sisi has become a bigger dictator than Mubarak…..
The president spoke in these meetings about Egypt's keenness in promoting democracy and protecting human rights, stating that Egypt is undertaking great efforts to respect civil and human rights, while strengthening security and stability as well.
Speaker Paul Ryan released a statement after his meeting with Al-Sisi, saying that "the strategic partnership between the United States and Egypt is essential to securing regional stability and defeating terrorism." He also added that the congress is keen to cooperate with its Egyptian counterparts during the upcoming period.
Again the US will accept authoritarian rule as long as their is a proxy in there to help with the US hegemony….respecting human rights while throwing protesters, journalists and bloggers in prison…..
Then there is the Turkish president and his referendum recently…..basically the referendum was an assault on rights of the people…..seriously limiting democracy in the country….and Trump was the first to call Erdogan and congratulate him on his win…..
Trump has acted according to type in singling out these categories, too, for his hostility, to the point of calling the press "enemies of the people" and calling for the imprisonment of his political rival, Hillary Clinton, an idea he has returned to from time to time.Turkey shows us what can happen when rhetoric passes to action. Let's take the example of the media and higher education. Over 100 news sites have been closed, and over 100 journalists have been imprisoned; Turkey is now the country with the highest number of confined journalists in the world. Many university professors, accused of anti-government sentiments or insulting the leader, have been escorted out of their homes and workplaces in handcuffs, as though they are criminals — which, for Erdogan, they now are.
President Trump called Philippine leader Rodrigo Duterte and expressed his commitment to their alliance and in developing "a warm, working relationship," a Filipino official said Sunday. Presidential spokesman Ernie Abella said Trump mentioned he was looking forward to visiting the Philippines in November to attend an East Asia summit and that Trump invited Duterte to the White House, reports the AP. "The discussion … was warm, with President Trump expressing his understanding and appreciation of the challenges facing the Philippine president, especially on the matter of dangerous drugs," Abella said. A White House statement described Saturday's call as "very friendly" and said the US-Philippine alliance "is now heading in a very positive direction." The remarks reflect a departure from Duterte's antagonistic stance toward former President Obama, whom he once told to "go to hell."
During Obama's final months in office, the Philippine president moved to build closer ties with China and Russia while threatening to end his nation's longstanding alliance with the US. On Sunday, three Chinese navy ships were welcomed in Davao city, Duterte's hometown, by his daughter, city Mayor Sarah Duterte. Abella said without elaborating that North Korea also came up in Trump's talk with Duterte. CNN notes that Duterte suggested earlier Saturday that the Trump administration should back away from standoff with North Korea to avoid risking a nuclear holocaust that could smother Asia. "It would be good for America to just restrain a little bit and if I were President Trump, I'll just back out, not really in surrender and retreat, but just to let the guy realize that, 'Ah, please do not do it.'" Duterte said. "It behooves America, who wields the biggest stick, just to really be prudent."
Duterte is a murderous little toad…..I do not think the America needs friends like him…….we can do so much better.
For the love of the "Little Dictator"……why are these people high on the "approval" list for our president?
Have an opinion? Then let it fly.


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Study Confirms All Poker Pros Are A Bunch Of Luckboxes
LAS VEGAS – In a shocking turn of events, a statistical study has confirmed that every single poker professional is a freaking luckbox. None of the so-called "expert plays" that professionals perform made the slightest bit of difference in the long run.
"We were a bit surprised," said Jay O'Connor, a UNLV statistician who co-authored the study. "However, these findings jibe with anecdotal reports from many others in the poker community, particularly dealers."
The study analyzed thousands of poker tournaments and their outcomes, paying close attention to tournaments in which a known professional went deep. Industry observers believe that this study is the first academic paper to refer to someone being "sodomized by the deck" and "having a horseshoe jammed up their [rectums]."
Pushback from pros
The findings are causing a rift at the table. Top professional Fedor Holz found the findings ludicrous.
"I work very hard at perfecting my game," he said from another final table. "My preparation is why I always seem to get cards, not some mystical luck of the draw."
Mr. Holz then proceeded to suck out on the river and knock out yet another competitor, adding to his mountainous chip stack.
Ironically, the dealer at Holz's table was one of the key interviewees for Professor O'Connor's study. Joe Fontana, 41, had plenty to say about poker millionaires and his own lost hopes.
"I've seen so many of these guys play, and they always have the hand," he said. "If I ran like that, I wouldn't have to wear this sweaty outfit every night and get cards thrown at me."
As he said this, pro Brian Rast hit a bona fide one-outer to take down a huge pot.
When asked about the hand, he merely shrugged and called it standard. For him, it probably is.
Also read: Several 2nd Place Finishers Come Forward With Chip-Dumping Allegations Against Mike Leah, Hashtag #IChoppedtoo Trends
Article credit: Bart Shirley


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As a first step in setting out how this argument will proceed, let us note a crucial ambiguity in Kerr's conclusion from the propositions of the 'nouvelle theologie' that the 'God exists' of the philosopher's reason 'means something radically different' from the 'God exists' affirmed by the Christian 'under the conditions of faith'. This is partly, but only partly, true, and to see in what sense it is true and in what sense false, we can ask: why does the Vatican Council, in distinguishing what it calls 'two orders of knowledge', distinguish them not only in respect of their source - the one being the product of reason, the other of divine faith and revelation - but also in respect of their 'object'? The question matters, for long before the 'nouvelle theologie' - at least since Pascal - there has been a quite generalised scepticism abroad whether, even supposing you could demonstrate a 'God of reason', that God of reason could be demonstrated to be the same God as the 'God of faith'.24 The answer to that question lies in the council's implicit reliance upon an ancient scholastic distinction between the 'material' and the 'formal' objects of knowledge: we can be acquainted with the same material object by sight and by touch; but sight acquaints us with it in respect of its colour, touch in respect of its sensitivity to temperature; so what they acquaint us with is the same thing materially -I see what is warm, I feel what is red - but differing formally: it is not as warm that I see it, not as red that I feel it. Hence, the knowledge they yield is in either case determined by its formal object, the material object being the same for both.
24 'Le Dieu des Chrestiens ne consiste pas en un Dieu simplement autheur des veritez geometriques et de l'ordre des elements; c'est la part des Payens et des Epicuriens . . . Mais le Dieu de l'Abraham, le Dieu d'Isaac, le Dieu de Jacob, le Dieu des Chrestiens, est un Dieu d'amour et de consolation.' Blaise Pascal, Pensées, ed. H. F. Stewart, London: Routledge & Kegan Paul, 1950, pp. 6-8.
This is not a wholly implausible way of construing the relationship between the God of the philosophers and the God of faith - the same God can be known under different descriptions, as 'warm' and 'red' are, and within different relations of knowing, as touching and seeing stand in differing relations of immediacy to their objects. And in fact the analogy with the different formal objects of the senses has, within the history of the subject, been employed directly, especially in the late Middle Ages. Giles of Rome, who (rather unconvincingly) thought of himself as a disciple of Thomas Aquinas, explained that the God of the philosophers is known as it were 'by sight', and the God of the theologians by 'touch' and 'taste'; for the philosophers know God 'at a distance' and intellectually across a gap crossed not by means of direct experience but by means of evidence and inference, and so through a medium, as sight sees; whereas, through grace and revelation, the theologian is in an immediate and direct experiential contact with God, as touch and taste are with their objects - touch and taste being analogies for the immediacy of love's knowledge.25 There is something to be said for this way of construing the relationship between the 'God of the philosophers' and the 'God of faith', for to do so is at least to acknowledge that the manner in which an object is perceived - the cognitive relation to it in which one stands - is determined by the descriptions under which it is perceived, while allowing that what is perceived in either case is one and the same object. As the philosophers say, the descriptions under which an object is perceived may be 'intentionally' distinct but 'extensionally' equivalent: the Morning Star is the same star as the Evening Star, though 'Morning Star' does not mean the same as 'Evening Star'. So it is, on Giles's analogy, with the natural and revealed knowledge of God.
Not every theologian, however, could have welcomed Giles's polarisation of philosophical detachment - 'seeing' - in opposition to theological experientialism - 'touching' and 'tasting' - and Thomas Aquinas nowhere does, providing us with a probably more helpful, because less polarised, account of sameness and difference of 'object'. What I see at a distance is a dark patch I can distinguish as a human being moving towards me. When it is close enough to me, I can see that it is Peter. When the object was at a distance what I saw was Peter, but it was not as Peter that I
25 'If we wish to speak of the contemplative life in terms drawn from the senses, we could, in a manner of speaking . . . say that the contemplation of the philosophers gives delight to hearing and sight; whereas the spiritual contemplation of the theologians gives delight to taste, smell and touch.' See my Eros and Allegory: Medieval Exegesis of the Song of Songs, Kalamazoo: Cistercian Publications, 1995, p. 364. I have translated this passage from Giles's text, misattributed to Thomas Aquinas, in the Venice edition (1745) of Thomas's Opera Omnia I.
saw him. Thus the God of reason in relation to the God of faith.26 The God the philosopher knows is the God of Abraham, Isaac and Jacob and of Jesus Christ; but the philosopher cannot, otherwise than by the reports of faith, know her God as the God of faith.27 This is the meaning of that famous, and famously derided, formula which Thomas Aquinas appends at the end of each of his 'five ways': et hoc omnes dicunt Deum.28 As Thomas concedes, the proofs of God prove very little indeed, but just enough: as 'proofs' they fall into that class of'demonstrations' which merely show that something exists by way of explanation (demonstratio quia), from which, no doubt some properties are derivable which must hold true of whatever thus far explains. But they are not explanations of 'effects' by way of what we demonstrate about them from the nature of their cause (demonstratio propter quid)29 because in any case (as we shall see30) we do not and cannot know the nature of God, we do not know what God is. Haldane explains:31 we can know from the fact that the water pressure to my shower is lower than in the rest of the system that there is a blockage in the inflow pipe to my shower-head. But just because I do not thus far know that what is obstructing the water supply is a small piece of masonry, as the plumber later discovers, it does not follow that what I know as 'blockage' is not what the plumber discovers to be a small piece of masonry, even though 'blockage' and 'small piece of masonry' do not mean the same. In parallel it should not be supposed that, having demonstrated the existence of a 'prime mover' or of a 'necessary being', Thomas imagines that 'all people' know God under such descriptions, still less that they worship God under such descriptions, even less still that they could love God under such descriptions. For this reason it is undoubtedly true that, as Kerr says, the 'God exists' of the philosopher does not mean the same as the 'God exists' known under the conditions of faith. And of course, in affirming that the God of his 'five ways' is what all people call by that name, he is by no means affirming that they do mean the same. The Latin et hoc omnes dicunt Deum should be translated not as 'this is how all people speak of God' or even that 'this is what all people mean when they speak of God', for manifestly they do not, and Thomas knows this: it should rather be translated as 'and this is the God all people speak of. The descriptions of the philosopher and of the ordinary believer are, as I have put it, extensionally equivalent; but of course they do not mean the same thing. How, then, do we know that these 'Gods' are extensionally equivalent, are one and the
26 ST la q2 al ad1. 27 ST Ia q2 al ad 1. 28 ST la q2 a3 corp.
31 J. J. C. Smart and John J. Haldane, Atheism and Theism, Oxford: Blackwell, 1996, p. 143.
same God? Only by faith: reason alone could not know that - it is the plumber, after all, not I, who knows that the blockage is a small piece of masonry.
Therefore, all the decrees of the first Vatican Council quoted above are statements of, or articulations of statements of, faith alone, for 'human beings are totally dependent on God as their creator and lord, and created reason is completely subject to uncreated truth'. So much by way of initial clarification.
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After producing two playoff teams last season, the AL West is back with a World Series favorite and a few teams looking to surprise. Oakland came out of nowhere last season to capture a Wild Card spot as the Astros once again won the division crown and returned to the ALCS. Houston enters 2019 as […]
Category: MLB Previews 2019
NL West Preview
The NL West was provided a bit of a shake up this offseason with Manny Machado heading to the San Diego Padres in free agency. This was a clear signal the Padres are serious about winning and will be a team on the rise starting now, but will it be enough to make the playoffs, […]
NL East Preview
The NL East looks a lot different than it did a year ago. Bryce Harper switched teams as the Phillies went all in, but does that make them the favorite to win the division?
AL East Preview
The AL East has the reigning World Series champs, but can they do it again? Or will the Yankees top their biggest rival for the division crown in 2019?


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DALLAS – As Muslim literature grows globally, a new publishing house is offering Muslim authors a rare chance to publish their fiction and afford readers the opportunity to experience diverse voices.
"I launched Djarabi Kitabs Publishing in an intent for my self-publishing works at the time to look more professional and avoid being censored by the industry," the company's founder, Papatia Feauxzar, told AboutIslam.net.
Feauxzar further explained that she expanded her publishing house to Muslim authors encountering similar issues.
According to its founder, Djarabi Kitabs Publications promises to infuse more diversity and provide opportunities for Muslim authors. It joins a growing list of Muslim publishers seeking authors who write about Muslim experiences.
It came to provide a solution to many Muslim authors who have a constant challenge in finding publishers willing to publish Muslim fiction.
During the Muslim Fiction and Mainstream Publishing Twitter chat, Muslim authors expressed how the lack of diversity in the industry, even in Muslim publishing, limits opportunities to publish their books.
Injecting Diversity and Inclusion
Muslim authors frequently run into obstacles to have their work published.
A major issue is that the publishing industry confines what is considered "Muslim" to specific parameters of exoticness and foreignness, resulting in a cognitive dissonance when reading works of Muslim authors conveying stories outside of those parameters.
"As an American Muslim author, I have had a challenging time finding publishers who value and celebrate a Muslim voice that is neither "foreign or exotic," Sahar Abdulaziz told AboutIslam.net.
Right now, the attention is on immigration stories or stories that present Islam or Muslim women as something exotic and "other."
Exposing the world to different Muslim authors is one of the objectives of the new Muslim publisher.
"Muslims are diverse and so are their art forms," Feauxzar told AboutIslam. "Therefore, I'm accepting any genre of writing."
Brave New Publishing
Another obstacle for some Muslim authors is finding Muslim publishing houses willing to print and distribute novels with certain content.
There is a divide in American Muslim fiction about including explicit content or hard hitting issues. Many Muslim authors with novels containing societal issues like domestic violence, sexual assault, incest, child sexual assault, terrorism and racism as well as the supernatural find it difficult to find a Muslim publisher willing to take on their work.
Aversions to sexual content are even stronger that to violence and gore among Muslim fiction authors and publishers.
Feauxzar told AboutIslam that Djarabi Kitabs is willing to push the envelope with authors and consider well-written novels with typically problematic content.
"I want my publishing house to stay true to the writer's voice and yet be polished and outstanding. I will make sure that each Muslim voice I publish will not be censored to please the reader. If a subject makes readers and other publishers shy, Muslim writers can trust and rely on Djarabi Kitabs Publications to get their work out there so we can have the open and needed discussions about these rampant issues."
Abdulaziz, known for controversial content in her novels, told AboutIslam that she appreciates Djarabi Kitabs' bravery and willingness to distribute novels with contentious plots.
"The controversial topics [in my books] might upset the status quo–shake the established apple cart. Many Muslim publishers equally shy away from anything controversial or provocative. However, Djarabi Kitabs Publishers immediately embraced the potential for controversy as long as the intention of the work is to shed light on the truth. This is an important distinction and a wonderful addition to our Muslim shelf life. I am proud to be a part of the team."
Muslim Authors at Home with Their Own
Djarabi Kitabs has a lineup of Muslim authors happy to have a publishing house that understands their experiences.
Social disconnect can be challenging for authors, editors, and publishers to produce a polished book that honors the author's voice.
Djarabi Kitabs' new author Zaheera Jina expressed appreciation for being under the roof a Muslim publishing house.
"The process becomes easier because we relate and understand the finer details of religion."


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Nitrogen Generator Systems
Nitrogen generator systems are critical in ensuring that vessel cargo is safely transported under a protective nitrogen padding system to prevent potential chemical reactions and possible ignitions of fire.
Hence, it is important to maintain N2 systems on vessels regularly to ensure smooth operation and immediate rectification when needed. As N2 system specialists, we are proud to represent two major marine N2 system makers as their authorised after-sales service centre in Singapore.
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Since Air Products and GIANTECH inked a service agreement in 2012, Air Products' customers have been able to enjoy convenient service and prompt technical support from GIANTECH here in Singapore.
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As part of this initiative, GIANTECH offers a Health Check Attendance Service (HCA) to vessels using Air Products' N2 systems and TMC air compressors. This annual maintenance plan aims to provide regular maintenance checks and prevent unnecessary system downtime. The ultimate aim is to provide quality services, so as to provide peace of mind to ship managers.
GIANTECH also keeps a ready stock of common running spares in our Singapore warehouse, including sensors and filter elements. We also supply membrane replacements for Air Products' N2 Systems.
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Wilhelmsen Technical Solutions provides marine equipment and engineering services to global markets at cost-efficient prices. Their main objective is to supply effective and safe solutions to cargo carriers. Constantly seeking to better meet customer demands and expand its services, Wilhelmsen Technical Solutions acquired Unitor to provide reputable and trustworthy N2 systems. As part of continuing efforts to grow the brand's capabilities, Wilhelmsen Technical Solutions has joined the Survitec Group in 2018.
GIANTECH's ongoing collaboration with Maritime Protection for inert gas systems led to the introduction of GIANTECH to Wilhelmsen Technical Solutions, and now, the Survitec Group. Consequently, both parties saw an opportunity for a partnership since GIANTECH also has experience and knowledge of N2 systems which Unitor specialises in.
Since 2013, GIANTECH has been undergoing intensive training under Wilhelmsen Technical Solutions – Unitor to enhance their technical skills and knowledge, with GIANTECH's engineers attending practical onboard training sessions with Unitor engineers. By undergoing regular training and staying up to date with Unitor's N2 system, GIANTECH constantly enhances its capabilities with regards to servicing Unitor products.
With strengthened competence, GIANTECH now provides the professional Health Check Attendance Service for vessels fitted with the Unitor N2 System. This cost-efficient, preventive measure aims to prevent unexpected and serious damage to vessels.
GIANTECH supports/provides:
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The 2nd major equipment, Kashiwa Nitrogen Gas Generator System, supplies extremely clean inert gas to prevent explosion in vessel cargo tanks. In collaboration with MEDAL, L.P (USA) and other membrane manufacturers, Kashiwa can supply systems of various capacity to match the flag and class requirements of different vessels.
Additionally, Kashiwa also supplies Marine PSA Nitrogen Generator in conjunction with KURAREY Chemical Co., Ltd.
GIANTECH also provides spare parts by Kashiwa to vessels trading in Singapore, Malaysia, India and many other global locations.
GIANTECH supports/provides:
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text: December 26, 2017Photo Story: Farolito Walk on Canyon Road, Santa Fe, NM Scroll down to content Share this:ShareTweetPrintEmailLike this:Like Loading... Related Published by BreCox Nutshell - Art Therapy/Counseling Grad Student, Board Member on New Mexico Art Therapy Association, Published Photojournalist/Writer, SCA member in the Kingdom of Outlands, Papercrafting/FSJ Coach, Air Force Veteran, Alum of Northern Kentucky University, INFJ, Progressive, Tech Geek, Dog Mom, Homegrown Ky girl Hello! Thanks for stopping by. Pull up a chair and stay a while. View all posts by BreCox


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A group of peaks in the north-western park of Nepal, called the Kanjiroba Himal, has been the obsession John Tyson for over 40 years. A modest school master, by profession, he returned to the Himalayas, again and again. His quest was to venture where no one had dared to explore in order to map uncharted territory.
The risk of such exploits can be great, even leading to the ultimate price. However, in 1961 John faced these risks in an expedition to this remote region. In 1998 at 70 years old, he returned to the peak of Kanjiroba Himal, for the last time.


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Trust McKinney Buick GMC for quality service, maintenance, or auto repair in TX. From routine maintenance to larger repairs, nobody knows your vehicle better than our Certified Service experts. Schedule an appointment today to take advantage of our current service offers on oil changes, brake repair, batteries, tires, and more.


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Season 2 of Stranger Things on Netflix was a joy to watch. It's always a tricky challenge to make the second season of a beloved show as good as the first season. It's even more difficult to make it better. I'm not saying season 2 is absolutely better (season 1 was such a fun ride that it will always edge out the rest), but in my opinion, it's as good as the first one and better in a couple of places.
The new season feels like it allowed every character it's moment to shine, even with the addition of a couple of new characters. We got to see the core group of kids as individuals with their own unique family lives and quirks. The scenes with Steve and Dustin were really fun, Eleven got to have a massive adventure and Hopper was again the anchor of the cast, and his little dancing scene has gone viral as hilarious memes…
Jim Hopper dancing to You Make My Dreams - Daryl Hall & John Oates #StrangerThings @noah_schnapp https://t.co/DSTJYL1jMj—
Hopper Dancing To (@hopperdancingto) October 29, 2017
I could go on and on…Paul Riser is always fun to watch…Sean Astin plays the role of "red shirt" Bob perfectly and the conspiracy theory character made me laugh out loud…I'll wrap it up by saying I highly recommend it. As a final note, here's the awesome soundtrack to the series, which will take you back to the mid-eighties in style.


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T-cell-mediated immune effector mechanisms play an important role in the containment of human immunodeficiency virus/simian immunodeficiency virus (HIV/SIV) replication after infection. Both vaccination- and infection-induced T-cell responses are dependent on the host major histocompatibility complex classes I and II (MHC-I and MHC-II) antigens. Here we report that both inherent, host-dependent immune responses to SIVmac251 infection and vaccination-induced immune responses to viral antigens were able to reduce virus replication and/or CD4+ T-cell loss. Both the presence of the MHC-I Mamu-A*01 genotype and vaccination of rhesus macaques with ALVAC-SIV-gag-pol-env (ALVAC-SIV-gpe) contributed to the restriction of SIVmac251 replication during primary infection, preservation of CD4+ T cells, and delayed disease progression following intrarectal challenge exposure of the animals to SIVmac251 (561). ALVAC-SIV-gpe immunization induced cytotoxic T-lymphocyte (CTL) responses cumulatively in 67% of the immunized animals. Following viral challenge, a significant secondary virus-specific CD8+ T-cell response was observed in the vaccinated macaques. In the same immunized macaques, a decrease in virus load during primary infection (P = 0.0078) and protection from CD4 loss during both acute and chronic phases of infection (P = 0.0099 and P = 0.03, respectively) were observed. A trend for enhanced survival of the vaccinated macaques was also observed. Neither boosting the ALVAC-SIV-gpe with gp120 immunizations nor administering the vaccine by the combination of mucosal and systemic immunization routes increased significantly the protective effect of the ALVAC-SIV-gpe vaccine. While assessing the role of MHC-I Mamu-A*01 alone in the restriction of viremia following challenge of nonvaccinated animals with other SIV isolates, we observed that the virus load was not significantly lower in Mamu-A*01-positive macaques following intravenous challenge with either SIVmac251 (561) or SIVSME660. However, a significant delay in CD4+ T-cell loss was observed in Mamu-A*01-positive macaques in each group. Of interest, in the case of intravenous or intrarectal challenge with the chimeric SIV/HIV strains SHIV89.6P or SHIVKU2, respectively, MHC-I Mamu-A*01-positive macaques did not significantly restrict primary viremia. The finding of the protective effect of the Mamu-A*01 molecule parallels the protective effect of the B*5701 HLA allele in HIV-1-infected humans and needs to be accounted for in the evaluation of vaccine efficacy against SIV challenge models.
The rate of human immunodeficiency virus type 1 (HIV-1) infection in developing countries has significantly increased in the last few years, and there is therefore an urgent need for the development of an effective vaccine. Studies in animal models have demonstrated the potential role of HIV-based neutralizing antibodies (Ab) in protecting against HIV infection (4, 34). The induction of neutralizing Ab against primary isolates by various vaccine modalities, however, has proven to be difficult, and restriction of viral replication by cell-mediated immune effector mechanisms appears to be a more realistic goal at present. The contribution of cell-mediated immune responses in controlling HIV-1 replication has been inferred in both acute and chronic HIV-1 infections (8, 27, 40, 42, 47) and clearly demonstrated in the simian immunodeficiency virus macaque (SIVmac) model (22, 35, 52).
Poxvirus-based vaccine candidates with various degrees of attenuation are known to induce cell-mediated immune responses and have been shown to prevent infection following challenge exposure to viruses with low virulence, such as some strains of HIV-2 (1, 3, 15) or SIVmac (21). They were also found to reduce viral burden following challenge exposure to highly pathogenic SIVmac isolates (7, 17, 41, 53). In addition, among the poxvirus vaccine candidates, NYVAC and ALVAC have also been demonstrated to be able to induce virus-specific CD4+ and CD8+ T-cell responses in SIV-infected macaques treated with antiretroviral therapy (18; our unpublished results).
ALVAC (canarypox virus)-based immunogens have been extensively evaluated as veterinary and human vaccine candidates (44) (unpublished results) with three such vaccines being registered with regulatory agencies. These are ALVAC-rabies and ALVAC-feline leukemia virus vaccines for cats and ALVAC-canine distemper virus vaccine for dogs. ALVAC-based HIV-1 vaccine candidates have been tested in more than 1,200 human volunteers and have been shown to be safe and immunogenic (11, 45). Preventive immunization of macaques with a canarypox vector-based HIV-2 immunogen was found to protect macaques from a nonpathogenic HIV-2 challenge (15). The relative efficacy of this vaccine modality, however, has not been assessed previously in the highly pathogenic SIVmac251 macaque model, in which disease progression and survival can be evaluated.
The usefulness of the SIVmac251 model in the evaluation of vaccine immunogenicity has been further enhanced by knowledge of macaque major histocompatibility complex (MHC) Mamu-A*01 status and SIV-specific epitopes restricted by this allelic form (2, 28–30). A study was therefore designed to assess whether immunization with an ALVAC-based vaccine candidate expressing the SIVmac251 Gag, Pol, and Env components and subsequent boosting with subunit gp120 boost could confer immunity and prevent or contain SIVmac251 replication following a mucosal exposure to SIVmac251. The results indicate that vaccination with ALVAC-SIV-gpe modified significantly the natural course of SIVmac251 (561) infection in Mamu-A*01-negative macaques (i.e., delayed the CD4+ T-cell loss) and that some Mamu-A*01-positive macaques naturally controlled viral replication. The MHC class I (MHC-I) Mamu-A*01 effect was also investigated following intravenous challenge with SIVmac251 (561) and SIVSME660 as well as with two simian/human immunodeficiency virus (SHIV) strains. The results indicate that the route of challenge exposure to SIV isolates influences the natural restriction of viremia in Mamu-A*01-positive animals regardless of the strain used and that in macaques infected with two independent SHIV isolates viremia restriction does not occur regardless of the challenge route.
Vaccines, immunization protocol, and challenge virus stock.Sixty-four macaques were used in this study. The ALVAC-SIV-gpe (vcp180) was engineered to express the gag, pol, and env genes of SIVmac251(K6W) (14) from the I3L and the H6 promoters (43). The H6 env and the I3L gag and pol cassettes were inserted in the ALVAC C3 locus in a head-to-head (5′-to-5′) configuration. Prior to amplification and purification of the vcp180 virus, the expression of the SIVmac251 genes was assessed in chicken embryo fibroblasts (data not shown). Groups A and B received 108 PFU of the ALVAC-SIV-gpe vaccine candidate by the intramuscular route at weeks 0, 4, 26, 52, and 113 or 143, and only animals in group B were inoculated intramuscularly with gp120 at weeks 26, 52, and 113 or 143 (Fig. 1). Groups C and D received 108 PFU of ALVAC-SIV-gpe gp160 at the same intervals as groups A and B by the intramuscular and, in addition, the intrarectal and intranasal routes (Fig. 1). Group D also received SIV gp120 (300 μg) adjuvanted in QS-21 (100 μg) at the same time intervals as group B (Fig. 1). The first control group received 108 PFU of ALVAC parental virus by the intramuscular route (group E), and the second was constituted of naive animals (group F). Group G received inoculations of gp120 (300 μg) in QS-21 adjuvant (100 μg) at weeks 0, 8, and 55. The SIV gp120 used for immunization was purified from the serum-free culture supernatant of SIVmac251 chronically infected Hut 78 cells by immunoaffinity column chromatography using anti-gp120 Ab as described previously (25). The macaques were challenged at week 117 of 147; animals were challenged intrarectally with 30 mucosal infectious doses of the SIVmac251 (561) isolate. The SIVmac251 challenge stock was prepared by culturing phytohemagglutinin (PHA)-activated peripheral blood mononuclear cells (PBMC) from a Mamu-A*01-positive infected macaque (561L) exposed to SIVmac251 by the vaginal route. The SIV challenge stock [SIVmac251 (561)] was titered in vivo in rhesus macaques by inoculating six animals with different dilutions of virus stock via the rectal route. Since six of six animals inoculated with the virus (0.5 ml diluted to 1.5 ml with RPMI medium) became infected, as evidenced by high plasma viremia and a drop in CD4 counts, this dose of virus was selected for all challenge studies.
SHIVKU2 challenge virus stock.The SHIVKU2 challenge stock (23) was prepared by culturing PHA-activated PBMC from an infected macaque that was inoculated intravenously with SHIVKU2. The SHIVKU2 challenge stock was titered in vivo in rhesus macaques by rectal inoculation of animals with different dilutions of virus stock. Since six of six animals inoculated with the virus (0.5 ml diluted to 1 ml with RPMI medium) became infected, as evidenced by high plasma viremia, this dose of virus was selected for all challenge studies.
Immunological assays.Serum samples were tested for SIV-specific Ab responses using an enzyme-linked immunosorbent assay (ELISA) described elsewhere (7). Serum titers were determined as the highest dilutions of immune serum producing ELISA values (A450) greater than or equal to two times the binding detected with a corresponding dilution of preimmune serum.
To assess SIV-specific serum-neutralizing activity, two types of assays were conducted with sera from the vaccinated animals. In the first assay, sera were tested for their ability to neutralize a T-cell-line-adapted stock of SIVmac251 grown in H9 cells and assayed in CEMx174 cells as described previously (38). In the second assay, neutralization of the challenge stock of SIVmac251 (561) was examined in PHA-activated human PBMC by measuring a reduction in viral p27 Gag antigen synthesis (31). CD4+ T-cell count in the PBMC of challenged animals was determined by standard flow-cytometric analyses (FAST Systems, Inc., Gaithersburg, Md.).
CTL assay and tetramer staining.PBMC (8 × 106) from macaques were cultivated in vitro with paraformaldehyde-fixed, autologous B-lymphoblastoid cell lines (B-LCL) infected with vaccinia virus encoding SIV Env and SIV Gag components. On day 3 of culture, 20 U of recombinant human interleukin-2 per ml was added to the cultures. On day 12 of culture, the lymphocytes were centrifuged over a Ficoll-diatrizoate gradient and assessed as effector cells in a standard 51Cr-release cytolytic assay. Target cells were B-LCL (106) cultured overnight with vaccinia virus encoding SIV Env, SIV Gag, or control antigen at a multiplicity of infection of 10 PFU/cell. B-LCL were then washed and labeled with 100 μCi of sodium 51chromate for 1.5 h. After being washed, 104 target cells were added per well in 96-well U-bottom plates in 100-μl volumes. Effector cells were added in another 100-μl volume at various concentrations to give effector-to-target ratios of 20:1, 10:1, 5:1, and 2.5:1. Plates were incubated at 37°C for 4 h. Fifty microliters of supernatant was transferred to counting plates and 200 μl of scintillation fluid was added and analyzed in a Wallac 1450 MicroBeta liquid scintillation counter. Specific release was calculated according to the following formula: (experimental release − spontaneous release)/(100% release − spontaneous release) × 100 (Table 1). The Mamu-A*01-positive rhesus monkeys were evaluated for p11C-specific cytotoxic T-lymphocytes (CTLs) using Mamu-A*01/p11C tetramer staining of unstimulated peripheral blood CD8+ T lymphocytes. Soluble tetrameric Mamu-A*01/peptide complexes were prepared, and 1 μg of phycoerythrin-labeled tetrameric Mamu-A*01/peptide complexes was used in conjunction with fluorescein isothiocyanate-labeled anti-human CD8α (Leu2a; Becton-Dickinson, San Diego, Calif.), energy-coupled dye-labeled anti-human CD8αβ (2ST8-5H7; Beckman Coulter, Fullerton, Calif.), and allophycocyanin-labeled anti-rhesus monkey CD3 (FN18) monoclonal Ab to stain p11C-specific CD8+ T cells. One hundred microliters of whole blood from the vaccinated monkeys was directly stained with these reagents, lysed, washed, and fixed.
Virological assay.Animals were bled periodically following challenge, and viral load in plasma was assessed using a nucleic acid sequence-based amplification assay to quantify SIV RNA (48). In addition, PBMC collected from animals 21 days following virus challenge were subjected to quantitative virus isolation by coculturing with CEMx174 cells to confirm virus transmission.
Study design.The experimental vaccination regimen included seven groups (A through G) of rhesus macaques (Fig. 1). The experimental groups A through D were immunized five times with ALVAC-SIV-gpe (108 PFU) and, at the time of the last three immunizations, animals enrolled in groups B and D were inoculated simultaneously with native SIVmac251 gp120. Control animals received either five immunizations with ALVAC vector (group E) or were left naive (group F). Lastly, group G animals received three inoculations with gp120. ALVAC-SIV-gpe was administered either by the intramuscular route (groups A and B) or by a combination of the intramuscular, intranasal, and intrarectal routes (groups C and D), whereas gp120 was administered by the intramuscular route, as described in Materials and Methods. A total of 19 Mamu-A*01-positive animals were included in the study and were distributed among the groups as summarized in Fig. 1.
Humoral immune response elicited by ALVAC-SIV-gpe.Serum Ab to SIVmac251 were measured in half of the animals from each group by ELISA using whole disrupted SIVmac251 virus spiked with purified gp120. Ab titers were negligible after two inoculations with ALVAC-SIV-gpe (data not shown). However, after the third immunization (week 26), the Ab titers increased and were highest in groups B and D, which also received the QS-21-adjuvanted gp120 (Fig. 2A). Although the Ab titers declined between immunizations, a steady level of Ab was present throughout the course of vaccination (Fig. 2A). A similar pattern was observed with the rest of the animals.
Both immunoglobulin G (IgG)- and IgA-specific Ab responses to SIV gp120 and p27 antigens were measured by ELISA in mucosal secretions such as rectal wash, saliva, and vaginal wash collected at week 115 in six animals from groups A through E. Neither IgG- nor IgA-specific Ab to SIV p27 were detected in animals from any of the vaccine groups (data not shown). In contrast, serum IgG-specific Ab response to gp120 was detected in most animals from groups B and D boosted with gp120 regardless of the route of administration of ALVAC-SIV-gpe. A low level of IgA-specific Ab to gp120 was detected in the saliva or vaginal secretions of two animals from group D only (data not shown).
Neutralizing Ab titers in sera from all ALVAC-vaccinated animals (groups A to D) collected at month 12 and 2 weeks following the final boost were measured against those of the laboratory-adapted SIVmac251 isolate in CEMx174 cells as well as the primary challenge stock of SIVmac251 (561) in human PBMC. For animals receiving the gp120 boost (group G), sera collected 2 weeks following the final boost were similarly assayed for neutralizing Ab titers. Neutralizing Ab titers to the laboratory-adapted SIVmac251 were detected in most animals receiving the vaccine candidate but were highest in animals boosted with the gp120 subunit preparation (Fig. 2B), whereas none of the serum samples neutralized the SIVmac251 (561) primary challenge stock in a human-PBMC-based assay (titers <1:5; 80% reduction in p27 synthesis was considered positive) (data not shown).
CTL activity induced by ALVAC-SIV-gpe.T-cell-mediated cytolytic activity was assessed in the blood of six of the immunized animals in each group as well as in control animals from group E following the third, fourth, and last immunizations. Cumulatively, 16 of 24 (67%) animals demonstrated an Env-specific CTL activity at at least one time point analyzed. Thirteen animals recognized Env target cells after the third immunization and, of those, nine were also positive following either the fourth or the fifth immunization (Table 1). Overall, 22% of the vaccinees demonstrated CTL activity against either Env or Gag at all times, 42% were positive two times, and 67% were positive at any single point. Among the control animals in group E, only one animal scored positive at a single time point. There was a trend suggesting that the relative frequency of measurement of CTL responses in peripheral blood in animals vaccinated by the systemic route was higher than in animals vaccinated also by the mucosal route (10 of 12 versus 6 of 12, respectively) (Table 1).
A surprising finding was that only 2 of 24 animals had detectable cytolytic activity against the SIV Gag protein, whereas 16 of 24 animals had cytolytic activity against the SIV Env protein. A possible explanation is that CTLs specific for Gag may have existed at a frequency below the level of detection by the assay. In fact, staining with tetramer for the peptide p11C epitope following specific peptide stimulation in vitro demonstrated that most of the vaccinated Mamu-A*01-positive animals (Table 1) indeed had memory CTL response to the Gag-immunodominant p11C epitope prior to viral challenge exposure (data not shown).
Mock-vaccinated Mamu-A*01-positive macaques naturally restrict SIVmac251 (561) replication.Following intrarectal challenge exposure to SIVmac251 stock 561, all 22 macaques from groups E, F, and G became viremic and seroconverted to viral antigens. Analysis of the plasma virus load during primary infection, set point, and chronic infection did not reveal significant differences among the groups, as summarized in Table 2. These data demonstrate that neither the gp120 subunit immunization nor the ALVAC mock vaccination influenced the virological outcome. Therefore, all these animals were gathered together to increase the statistical power of the subsequent analysis.
Interestingly, measurement of viral RNA in the Mamu-A*01-positive and -negative mock-vaccinated macaques revealed that among the Mamu-A*01-positive macaques four of five controlled viremia at set point up to week 17, as demonstrated for each animal in Fig. 3A. In fact, when the virus load in all the control animals (groups E, F, and G) was stratified according to the Mamu-A*01 status (15 were Mamu-A*01 negative and 7 were Mamu-A*01 positive), statistical analysis of the viral-load data demonstrated that in the seven mock-vaccinated Mamu-A*01-positive animals virus load was significantly lower than in the Mamu-A*01-negative macaques during primary viremia (0 to 28 days), at set point (2 to 3 months), and thereafter (for primary viremia, P = 0.0066 by the Wilcoxon rank sum test; for set point, P = 0.0007 by the Wilcoxon-Gehan test; for median viremia, P = 0.0068 by the Wilcoxon-Gehan test). In fact, most Mamu-A*01-positive animals had nondetectable viremia at set point and thereafter, as demonstrated for each animal in Fig. 3A and collectively in Fig. 3B. All together, these data indicate that the Mamu-A*01-positive macaques naturally restricted replication following intrarectal exposure to SIVmac251 (561).
Effect of ALVAC-SIV-gpe vaccination in Mamu-A*01-positive animals.Because of the natural ability of Mamu-A*01-positive macaques to control SIVmac251 (561) intrarectal infection, the statistical analysis of the relative efficacy of ALVAC-SIV-gpe vaccine was assessed independently in the Mamu-A*01-positive and the Mamu-A*01-negative animals. Genetic characterization of all animals enrolled in this study demonstrated the presence of 12 Mamu-A*01-positive animals among the 42 ALVAC-SIV-gpe-vaccinated macaques (Fig. 1). Unknown to us at the beginning of the study, the vaccinated Mamu-A*01-positive animals were interspersed unevenly in the experimental groups A through D (Fig. 1). Since in retrospect we observed a significant containment of viremia in the Mamu-A*01-positive animals (Fig. 3B), the overall evaluation of relative vaccine efficacy was assessed independently in Mamu-A*01-positive and -negative vaccinated and control macaques.
In Mamu-A*01-positive vaccinated and control macaques, the overall difference in viremia during primary infection (0 to 28 days) was not significant (Table 3), even though the vaccinated macaques appeared to control viremia faster than the nonvaccinated Mamu-A*01-positive animals (Fig. 4A). In fact, the quantitation of CD3+ CD8+ peptide p11C tetramer response in the blood of animals following challenge exposure demonstrated a faster appearance of this response in the vaccinated animals than in control animals (Fig. 4B), consistent with a secondary response. This difference was found to be significant in the interval from day 13 through day 28 postchallenge (P = 0.0075) by the application of repeated-measures analysis of variance to the arc-sine-transformed tetramer data (Fig. 4B). These data further support the importance of cell-mediated immunity in controlling SIV replication, as demonstrated by other studies (22, 29, 35, 52), and indicate that the ability of Mamu-A*01-positive animals to restrict viral replication may mask the relative efficacy of vaccines. No significant difference in the early CD4+ T-cell drop was observed between these two groups (data not shown). Accordingly, no deaths were observed in the Mamu-A*01-positive group regardless of the vaccination status (Fig. 5C).
ALVAC-SIV-gpe vaccination decreases primary viremia and CD4+ T-cell loss in Mamu-A*01-negative macaques.The effects of ALVAC-SIV-gpe vaccination on viral load and the natural course of SIVmac251 infection were evaluated separately in the Mamu-A*01-negative macaques. Of the 30 vaccinated and 15 mock-vaccinated Mamu-A*01-negative macaques, all became infected except 2 vaccinees. The 30 vaccinated animals experienced lower viremia in primary infection (first 28 days) than the 15 control macaques (P = 0.0078) (Table 3). When the peak viremia in the 30 vaccinated animals was compared to that in 11 control macaques (excluding 4 macaques that received gp120), this difference was even more significant (P = 0.0034) (Fig. 5A). The set-point viremias in the control and vaccinated macaques did not differ significantly (Table 3), but, remarkably, the analysis of the absolute CD4+ T-cell count (Fig. 5B) indicated that the 30 vaccinated animals were protected from acute loss of CD4+ T cells within the first 2 months of infection (P = 0.0099) and during the 1-year follow-up (P = 0.03). Among the vaccinated Mamu-A*01-negative macaques, fewer deaths occurred than among Mamu-A*01-negative control animals, but this difference did not reach statistical significance (Fig. 5C).
The contribution of boosting with the gp120 subunit preparation and the route of immunization of the ALVAC-SIV-gpe vaccine effect were also analyzed by comparing virus load and CD4+ T-cell count in Mamu-A*01-negative animals from groups A and C to those in animals from groups B and D. As demonstrated in Table 2, neither the monomeric gp120 subunit preparation nor the combination of the routes of immunization appeared to significantly contribute to the effect of ALVAC-SIV-gpe vaccination. Accordingly, no significant difference in the CD4+ T-cell counts was observed between the vaccinated groups of animals (data not shown).
Collectively, these data demonstrate that a decrease in virus load during primary and chronic infection and preservation of CD4+ T cells occurred in Mamu-A*01-positive animals regardless of their vaccination status. In Mamu-A*01-negative animals, however, a vaccine effect was also observed, and those macaques experienced a significantly lower level of viremia during primary infection than did control macaques. This effect was presumably associated with a better preservation of CD4+ T cells and was associated with fewer deaths in the vaccinated animals than in control animals within 1 year of SIVmac251 (561) infection.
Preservation of CD4+ T cells in Mamu-A*01-positive macaques following intravenous challenge with either SIVmac251 (561) or SIVSME660.We further investigated whether the inherent ability of Mamu-A*01-positive macaques to restrict viral replication observed in our study was dependent on the route of challenge. To do so, virus load and CD4+ T-cell counts were analyzed for up to 24 weeks postchallenge in 5 Mamu-A*01-positive and 12 Mamu-A*01-negative naive macaques previously infected intravenously with the same 561 stock of SIVmac251. In addition, to assess whether the Mamu-A*01 effect could also be extended to other SIV isolates, we retrospectively analyzed the virus load and CD4+ T-cell count in Mamu-A*01-positive and -negative control macaques infected previously with SIVSME660 (41, 53, 54). Analysis of the virus loads in the blood of macaques following intravenous exposure to SIVmac251 (561) or SIVSME660 did not reveal a significant reduction in virus load within the first 20 weeks of infection in animals from either group (data not shown). However, longitudinal analysis of the absolute CD4+ T-cell count during the same period indicated a better preservation of the absolute CD4+ T-cell count in Mamu-A*01-positive macaques infected with either virus (Fig. 6). In fact, in animals infected with the SIVmac251 (561) strain, repeated-measures analysis of variance on the square-root-transformed CD4+ T-cell count over a 6-month interval revealed that the differences in CD4+ T-cell counts were significant at the P level of <0.001 for each of the intervals analyzed (Fig. 6, top panel). Similarly, in macaques inoculated intravenously with SIVSME660, the loss in CD4+ T-cell counts differed significantly between Mamu-A*01-positive and -negative animals (Fig. 6, bottom panel) and was delayed in Mamu-A*01-positive macaques (weeks 8, 12, and 16; P < 0.052 for each by the Wilcoxon rank sum test and P < 0.0001 by the repeated-measures analysis of variance). All together, these results need to be accounted for in the evaluation of a vaccine effect when preservation of CD4+ T cells is included as a parameter of vaccine protection.
The Mamu-A*01 effect is not evident following infection with two chimeric SIV/HIV-1 strains.Other macaque models used in the evaluation of HIV-1 vaccine candidates include the use of chimeric SHIVs. To investigate whether the Mamu-A*01 molecule could also have a protective effect following SHIV infection, the viremia and CD4+ T-cell counts of Mamu-A*01-positive and -negative macaques following challenge with either SHIV89.6P by the intravenous route or SHIVKU2 by the intrarectal route were compared. Viremia levels and CD4+ T-cell numbers did not differ among Mamu-A*01-positive and -negative animals following SHIV89.6P intravenous challenge (Fig. 7, upper panels) or SHIVKU2 intrarectal challenge (Fig. 7, lower panels).
In this study, we have demonstrated that prophylactic ALVAC-SIV-gpe vaccination of macaques followed by intrarectal challenge exposure with the highly pathogenic SIVmac251 strain was associated with lower primary viremia, better preservation of CD4+ T-cell counts during primary and chronic infection, and prolonged survival. The neutralizing Ab response induced by a monomeric gp120 subunit preparation did not contribute to the observed protection from disease, presumably because none of the animals developed an Ab response capable of neutralizing the primary-challenge stock virus. However, vaccination with the ALVAC-SIV-gpe induced SIV-specific CD8+ T-cell virus-specific cytolytic activity in 67% of the vaccinated animals and was associated with a significant secondary response to the Gag p11C peptide in the Mamu-A*01-positive macaque following virus challenge exposure. Similarly, the vaccinated Mamu-A*01-negative macaque did better than control nonvaccinated macaques.
It is to be noted that the viral challenge stock used in this study infected 100% of the control macaques (22 control animals and 6 animals in the titration study). It is quite likely that the infectivity of this stock as well as the size of virus inoculum used during challenge may far exceed that of HIV-1 transmission, as the recently estimated frequency of transmission of HIV-1 in humans appears to be relatively low (46, 50).
The immunogenicity of ALVAC-based HIV vaccine candidates has been extensively studied in human volunteers enrolled in clinical trials of phases I and II, and the immunogenicity of this vaccine in humans mirrors that observed in the macaques studied here (6, 10, 12, 13, 16, 24, 51). However, despite clear evidence of immunogenicity, enthusiasm to proceed to extended efficacy trials has been tempered by incomplete knowledge of vaccine-induced immune parameters and protection against lentivirus exposure. The study presented here indicates that although vaccinated macaques were not protected from infection they did demonstrate significant differences in the virological and clinical outcome following exposure to a pathogenic SIVmac251 challenge. It should be noted that despite such uncertainties as to whether results obtained in the macaque model can be extrapolated to humans, recent studies seem to indicate that it may be appropriate to do so. For instance, therapeutic intervention in SIVmac251 primary infection (18, 20, 32, 33, 39, 55) has paralleled closely results obtained in primary HIV-1 infection in humans (49), validating the SIV macaque model. Furthermore, the observation presented in this communication (that Mamu-A*01-positive macaques are better able to control viral replication than macaques carrying other MHC-I molecules) highlights additional similarities to HIV-1 infection of humans. In HIV-1 infection, the maximum heterogeneity of HLA class I molecules or the single HLA-B*5701 molecule has been associated with a more benign clinical course (9, 37) whereas HLA A1-B8-DR3, B27, CW7, and B*35-Cw*04 molecules are correlated with faster disease progression (9, 26, 36, 56). Mamu-A*01-positive rhesus macaques appear to develop early robust multiepitope virus-specific CD8+ T-cell responses (2) (B. R. Mothe et al., submitted for publication), which persist in various compartments (19) and may contribute to their ability to restrict viral replication, as observed in this study, and this further underscores the importance of the breadth of CD8+ T-cell immune response to the containment of viral replication. Finally, this study indicates that the genetic background of macaques used in preclinical studies of the relative efficacy of the SIVmac251 model needs to be accounted for in comparative studies. In this context, it is to be noted that upon intravenous challenge the Mamu-A*01-related protective effect was more evident on the loss of CD4+ T cells than on viremia. This effect did not appear to be restricted to our viral stock, since it was observed also with the SIVSME660 strain. Interestingly, however, in other studies in which intravenous transmission of SHIV89.6P viruses was assessed in naive Mamu-A*01-positive and -negative macaques (5) and in our studies following intrarectal challenge with SHIVKU2, the Mamu-A*01 effect was not evident, regardless of the route of challenge. These differences between SIV and SHIV strains are unclear but may be explained by a differential response of rhesus macaques to HIV-1 proteins (Env, Tat, Nef, Rev) present in the SHIV chimeric virus. In summary, our findings demonstrate that the ALVAC-based SIV-gpe vaccine protected macaques from disease induced by this highly pathogenic virus and warrant more testing of the efficacy of an ALVAC-based HIV vaccine candidate in humans.
We thank David I. Watkins for helpful discussion, Sharon Orndorff for technical coordination, and Steven Snodgrass for editorial assistance.
This work was supported in part by National Institutes of Health grant AI-85343 (N. L. Letvin and D. C. Montefiori), National Institute of Allergy and Infectious Diseases contracts N01-AI55271 and N01-AI55260 to Advanced BioScience Laboratories, Inc., and National Institute of Allergy and Infectious Diseases contract NIH-NIAID-AI-65312 (M. G. Lewis).
- Received 29 May 2001.
- Accepted 3 October 2001.
- ↵*Corresponding author. Mailing address: National Cancer Institute, Basic Research Laboratory, 41/D804. Bethesda, MD 20892. Phone: (301) 496-2386. Fax: (301) 402-0055. E-mail: franchig{at}mail.nih.gov.
- American Society for Microbiology


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What is it?
Out with the old, in with the new. That's the message Mercedes-Benz is extolling with the SLC. Don't be fooled, though. Despite the change in name, the two-seat roadster is nothing more than a mildly facelifted version of the third-generation SLK, which first hit UK showrooms back in 2011. Not so new, then.
Read our full Mercedes-AMG SLC 43 review here
Among the subtle mid-life styling changes brought to Mercedes-Benz's compact roadster are a redesigned front bumper with larger ducts for improved engine bay cooling, an altered grille design and revised headlights with integrated LED daytime running lights and fresh internal graphics. At the rear, there are new LED tail-light graphics and a reprofiled bumper with integrated tailpipes and a more pronounced diffuser element.
Revisions to the operating procedure of the SLC's folding hard-top now allow it to be opened without having to manually set the luggage cover in place. It also now operates at speeds of up to 25mph, allowing it to be opened and closed while setting off rather than at a standstill, as with the older SLK. Buyers can continue to order the optional Magic Sky Control glass roof, which employs an electrochromic process to vary the amount of light entering the cabin.


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Finding a place to start a business venture is often a hurdle for aspiring entrepreneurs. Mahima P S, a budding entrepreneur from Ollur in Thrissur district, is proving that it's possible to overcome such challenges if you have imagination and confidence at your disposal.
The startup business
Mahima proposes to make available the services of people skilled in video editing, album designing, videography, and photography. Apart from taking up technical jobs at the studio, the startup will undertake field work for clients.
Why the business model
Mahima has an industry experience of over seven years in album designing. She is confident of the rapport that she is able to build with her clients; it has proven beneficial in her career where a clear understanding of the client requirements plays a crucial role, she says. Her husband is currently working as videographer in another company and can spare time for the startup as well. Mahima says she has always wanted to be an entrepreneur. This venture will give the additional satisfaction of providing job opportunities to others in the field, she says.
Capital required
The studio functions in a rented space of 350 sqft. The equipment cost including that of computers will come up to Rs 6.5 lakhs. Rolling out the services will incur an additional expense of Rs 50,000.
Facing Competition
Mahima is aware that her startup will face stiff competition in the industry which has countless well-established firms. She is confident that her skills in art work and the industry experience will come to her aid in facing the challenges. Her knowledge of the consumer base and people skills will be put to use for gaining foothold in the industry, she says. Liaisoning with agencies in Bangaluru and Chennai is already underway to ensure regular flow of work. The initial plan is to focus on undertaking work related to wedding videos. During the wedding seasons, the focus will be on field work and during off seasons, the studio work will be in full swing, she says.
Freshers are welcome aboard
Mahima says that experience in the profession is of little consequence in a startup like this. The co-ordination is where a little inside knowledge of the industry will be help, which she can manage on her own. The team working on the field could very well be made up of freshers with a creative bend of mind. Original thinking and creativity are the two vital aspects here, she says.
As per the industry rates, page designing and printing can be priced at Rs 400-700 depending on the quality. A client who requires page design alone will be charged Rs 70-120 per page. Mahima says that the projected returns should come to around four lakhs per month with a net income of Rs 80,000.
Mahima can be contacted at - M D Media, Ollur P. O., Kesavapadi, Marathakkara, Thrissur.


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4 Posts
Discussion Starter • #1 •
When I turn the key the fuel pump does not work.To get me home we lifted the rear seat cut to 2 wires.Connected to a 12volt battery brown to -ve, cotton covered to +ve.This made the fuel pump run all the time.Returned home reconnected fuelpump still not working. I have a 190E im breaking (engine dead fuel pump works fine),so swaped black fuelpump relay by battery.Still dead. The red relay with the 10Amp fuse could not be swaped as its a different type.Then one at a time swaped the two relays in the fusebox.The fuel pump still will not work HELP?? Please..


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The Test Case trailer: Nimrat Kaur plays an army captain in the upcoming ALTBalaji web series
Nimrat Kaur plays Captain Shikha Sharma in The Test Case directed by Nagesh Kukunoor and Vinay Waikul.
In the show, Shikha Sharma is the only woman to be enrolled in the Special Forces Training Centre. Her dream is to become the first female commandant of the force, but it turns out to be more difficult than she ever thought it could be. She meets her peers and seniors — all of whom are male — who mock her and call her "military barbie".
The environment at the camp is hostile, and she needs to prove herself worthy of being at there. We see her struggle to fight through military obstacle courses in an attempt to prove that women, too, can be an invaluable asset to the army. As the trailer reveals, something goes horribly wrong with Shikha while training to fight the enemy across the border. Things then take an interesting turn as military secrets starts getting leaked, and Shikha comes under the scanner.
Nimrat brings forward a strong female character in the Ekta Kapoor produced show with dialogues such as, "this is my story sir, and there is only one hero in this story, and that's me."
The Test Case, also stars Akshay Oberoi, Atul Kulkarni, Rahul Dev and Sumit Suri, and will be available on the streaming platform ALTBalaji from 26 January.
You can watch the trailer here.
Find latest and upcoming tech gadgets online on Tech2 Gadgets. Get technology news, gadgets reviews & ratings. Popular gadgets including laptop, tablet and mobile specifications, features, prices, comparison.
Updated Date: Jan 17, 2018 17:50:52 IST


text: 2 Sometimes we need simple reminders to keep ourselves in check. Summer brings outdoor fun, travel, vacations and sometimes long and busy days where we can easily slack off in the nutrition department. This is a dish I make in the summer with fresh ingredients from the local farmers market. Fresh […] Food Prep: Summer Fare25 Jun, 2015 in FUEL "recipes" tagged eating healthy / farmers market / Food / food choices / Food prep / healthy eating / healthy lifestyle / kale / mindful eating / protein / summer food / tomato by yesrising


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Sealing belt
A sealing belt made of laminated PTFE glass fabric and fitted with an even weld will ensure that your packaging foil is transported without sticking to the heat source. This allows for a continuous sealing process. But what exactly is a sealing belt and what properties should you pay attention to?
First of all: if you are in the market for a sealing belt, the best approach is contacting Hardick. Hardick specialises in customer-specific PTFE products and can definitely get you the sealing belt that you need. We have our own production facility and many of the raw materials in stock, so we can ensure you a swift delivery.
Sealing belt with non-stick properties
A sealing belt is specially designed to be used when direct heat is applied to seal products and packaging. If this heat seals the product, you don't want the product to stick to your belt. That is why it must have non-stick properties. If the products do stick to the belt, this will result in expensive and time-intensive delays.
Our sealing and PTFE conveyor belts are widely applicable. They have been successfully implemented in the production of food products, bake and fry products, and extruded rubber products for car industries. However, they are still mainly used in the packaging industry, due to their non-stick properties.
Excellent service
If you purchase your products from Hardick, you are not just getting the best quality. You can also count on the best service. In the past few decades, we have become widely known for our excellent service, which also plays an important part in our ISO 9001 certification. We'll help you whenever you need, with whatever you need.
This especially applies to emergencies. In case of emergency, you can contact our service team. This team is available 24 hours a day and 7 days a week. We'll find out what exactly is going on and we will help you right away. If a band is broken, for instance, we can produce a new one within hours.
Hardick is your partner for any sealing belt
Of course, we understand that you require a full-service partner. And that is exactly what Hardick is. With a global network of transporters, a large stock of high-quality products and experienced staff that has – for the most part – worked with us for over ten years, you can really count on our expertise.
Whether you are in the market for a sealing band or if you need PTFE welding equipment; we're here to help. Contact us through our website, send us an e-mail at firstname.lastname@example.com or call us. Our phone number is 0031 53 435 32 35. We'd love to tell you more about our products and services, and about what we can do for you.


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Going to School
Bihar and JharkhandSECTORS
We undertook the baseline evaluation of the impact of Going to School's 'Be! Schools' program which teaches skills to students across Bihar and Jharkhand.
Surveys were administered to students in Grade 9 in Rashtriya Madhyamik Shiksha Abhiyan and Kasturba Gandhi Bharatiya Vidyalaya schools in Bihar and students of Grade 7 and 8 in Kasturba Gandhi Bharatiya Vidyalaya, Tribal Residential and Congregation schools in Jharkhand. The study, a baseline evaluation of the outcomes of the client's intervention to promote development of skills in schools was conducted across 480 schools in 40 districts.
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I picked this foliage bouquet on Oct. 28. That night the overnight low matched the date: 28 degrees F. (-2.22 C. Thirty-two degrees is freezing under this system.) Since then we've had nights with temperatures in the single digits. But a number of these plants are still going strong in the garden. Strong enough that I have pulled a few things out as they faded and replaced them with fresh ones.
As I was taking these photos I realized this bouquet needed some brighter yellow-greens but I could not really find any left in the garden to cut.
The arrangement includes dramatic leaves like Hosta, Arum italicum and Hellebore along with lots of ferns and Heuchera. I also cut the last two stems of Alstroemeria 'Inca Ice' that had buds on them.
Those long brown seed pods are from my Locust trees and there is also a stem of Hosta flowers that have gone to seed.
Here's what this grouping looked like after two weeks of dry indoor air. I did replace the Amsonia fronds but my favorite foliage is the brown and crispy Hosta leaf.
Surprisingly the buds on the Alstroemeria actually continued to plump up and then open after being cut. The color is a bit paler than the ones cut directly from the garden in full flower, but otherwise they look pretty good. I'll keep this bowl of foliage going as long as possible. Usually it lasts a month or so with a few additions and subtractions along the way.
I updated this grouping one last time, adding the remaining Alstroemeria from this vase. Their faded color nicely matches that ones that were cut in bud. It would seem as though I have a Thanksgiving bouquet at the ready, just waiting for the table to be set!
To see how other gardeners put together a vase this morning, visit Cathy at Rambling in the Garden who hosts this most satisfying meme.


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As a Pennsylvania resident, you may cancel this transaction, without any penalty or obligation, within ten (10) days from the above date (the date you received this notice).
If you cancel, any property traded in, any payments made by you under the contract or sale, and any negotiable instrument executed by you will be returned within ten (10) business days following receipt by Bluegreen of your cancellation notice, and any security interest arising out of the transaction will be canceled.
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If you do make the goods available to Bluegreen and Bluegreen does not pick them up within twenty (20) days of the date of your notice of cancellation, you may retain or dispose of the goods without any further obligation. If you fail to make the goods available to Bluegreen, or if you agree to return the goods to Bluegreen and fail to do so, then you remain liable for performance of all obligations under the contract.
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