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There appeared to be a glimmer of hope for the restaurant industry last month, when despite ongoing negative restaurant sales and traffic performance in April, BlackBox Intelligence Executive Director, Victor Fernandez said that "there are some reasons to be cautiously optimistic about the second quarter, at least in terms of improvement over what we've seen in the recent past" adding that "the move of the Easter holiday meant that April's results were likely softer than they would have been without this shift, meaning spending in restaurants was probably a little stronger than the numbers show."
Alas, any trace of optimism was doused with the latest BlackBox snapshot report (based on weekly sales data from over 27,000 restaurant units, and 155 brands representing $67 billion dollars in annual revenue) which found that May was another disappointing month for chain restaurants by virtually all measures.
Same-store sales were down -1.1%, which represents a 0.1% decline from April. At the same time, same-store traffic "growth" also dropped by -3.0% in May, down 3.2% on a rolling 3 month basis. Although traffic results improved from prior month, the growth in check average was lower than it has been in recent months, causing the fall in sales growth vs. March and April.
More concerning is that the restaurant industry has not reported a month of positive sales since February of 2016, according to BlackBox.
The latest report from the National Restaurant Association found much of the same:as a result of softer sales and customer traffic levels and dampened optimism among restaurant operators, the National Restaurant Association's Restaurant Performance Index (RPI) registered a sizable decline in April. The RPI – a monthly composite index that tracks the health of and outlook for the U.S. restaurant industry – stood at 100.3 in April, down 1.5 percent from a level of 101.8 in March.
- The Current Situation Index, which measures current trends in four industry indicators (same-store sales, traffic, labor and capital expenditures), stood at 99.1 in April – down 2.3% from a level of 101.4 in March. April represented the sixth time in the last seven months with a reading below 100, which signifies contraction in the current situation indicators.
- The Expectations Index, which measures restaurant operators' six-month outlook for four industry indicators (same-store sales, employees, capital expenditures and business conditions), stood at 101.5 in April – down 0.7 percent from March. Although the Expectations Index remained above 100 – signaling the anticipation of generally positive business conditions in the months ahead – it declined to its lowest level in six months.
April's sharp decline in the RPI was the result of broadbased drops in both the current situation and expectations indicators. And, as BlackBox found, the Natl Restaurant Association confirmed that restaurant operators reported a net decline in same-store sales and customer traffic, which followed modestly stronger results in March. In addition, restaurant operators' six-month outlook for both sales growth and the economy retrenched from more positive readings in recent months.
Restaurant operators reported a net decline in same-store sales for the sixth time in the last seven months. Only 34% of restaurant operators reported a same-store sales increase between April 2016 and April 2017, down sharply from 57% of operators who reported higher same-store sales in March. 47% of operators said their sales declined in April, up from 30 percent who reported similarly in March.
Restaurant operators also reported softer customer traffic levels in April. Only 26% of restaurant operators reported an increase in customer traffic between April 2016 and April 2017, down from 41 percent of operators who reported higher traffic in March. Fifty-two percent of operators reported a decline in customer traffic in April, up from 38% in March.
* * *
Discussing the latest results, Fernandez said that "at this point, we believe the most likely scenario for the current quarter will be an improvement over recent quarters, while still suffering negative sales given the current consumer spending trends." Or, as we would put it, no actual improvement.
Looking at the macro picture, where there has recentlyy been an upturn in retail spending on most goods and services, adds to the confusion as it stands in stark contrast to the continued decline in sales growth at restaurants. Consumers appear to be maintaining their spending at restaurants - at a declining pace - but increasing it for other goods and services. This change in consumer spending patterns was identified about a year ago and how much longer it will continue is unclear.
Additionally, the restaurant operators' outlook for the economy is not as bullish as it was in recent months. 22% of restaurant operators said they expect economic conditions to improve in six months, down 15% points from the reading in December 2016. 12% of operators expect economic conditions to worsen in six months, while about two-thirds think conditions in six months will be about the same as they are now.
The details:
- May sales were weak across all segments. Only the fine dining segment was able to achieve very small positive same-store sales growth during the month. The second best performing segment during May was quick service. That soft performance notwithstanding, the best performing segments continue to be those with the lowest and highest average guest checks. "Dining experience on one end and value and convenience on the other seem to continue to be key components of restaurant sales performance based on current consumer spending trends," said Fernandez.
- The weakest performing segment in May was casual dining. This was a bit unexpected since the segment showed improved performance during the first four months of 2017 after lagging the industry for several years. Casual dining has added a modest number of new units, but same-store sales declines have contributed to its overall loss in market share.
- Despite weak sales results year-to-date, fast casual continues to win the market share battle. It gained the most share in the first quarter of 2017 compared with the same quarter a year ago. Aggressive expansion has driven total sales growth, but increased competition and market build-out have undoubtedly impacted same-store sales for the segment. The only other segment that gained market share year-over-year was quick service.
There is a silver lining: while overall sales continue to decline for most of the industry, there are pockets of opportunity that some brands have capitalized on to boost performance. Dine-in sales have been negative year-to-date, but to-go is up 2.9 percent, perhaps facilitated by recent introductions of smartphone-based ordering. Sales are also up in catering, delivery and drive-thru. From a day part perspective, breakfast and mid-afternoon sales offer continued opportunities for growth, while lunch and, especially, dinner sales continue to stumble.
Ironically, in addition to challenges from falling guest counts and consumer spending, strong challenges continue to confront restaurants in both staffing and retaining enough qualified workers. We say ironically, because as we showed after the latest jobs report, restaurant/fast food/waiter/bartender hiring remains the only strong spot in the US labor market. As the chart below shows, starting in March of 2010 and continuing through April of 2017, there have been 87 consecutive month of payroll gains for America's waiters and bartenders, an unprecedented feat and an all time record for any job category. Putting this number in context, total job gains for the sector over the past 7 years have amounted to 2.378 million or just under 15% of the total 16.4 million in new jobs created by the US over the past 87 months
And yet, according to BlackBox, restaurant operators are pessimistic regarding the difficulty of recruiting in the upcoming quarters. Part of the problem is that hiring for new restaurant positions has started to pick up again. The number of employees in the chain restaurant sector increased by 1.9 percent during April compared with a year ago, up from 1.5 percent growth recorded in March. The other issue affecting staffing is rising turnover. Turnover rates for both hourly employees and management staff increased again during April. "The turnover numbers that we are reporting are stunning", said Joni Thomas Doolin, CEO of TDn2K. "Many of the brands that we track are already facing unsustainable levels of staffing vacancies. Most alarming is the fact that over 70% of employees are leaving voluntarily as opportunities for better work increase".
Meanwhile the overall labor market nearing full employment doesn't hint at relief for operators any time soon. The consequences of turnover are well documented by TDn2K. Not only does it impact service levels and guest satisfaction, which correlate to traffic and sales, but it is also a huge source of additional costs hurting the bottom line. According to a recent study by People Report, it costs on average about $2,200 to replace a single restaurant hourly employee, while the cost of turnover for all levels of restaurant management is on average about $15,000 per manager.
"The companies who are leading in the marketplace are starting by winning in the workplace. Being a great employer has never been more important," stressed Doolin.
The summary: after 14 months of continuous declines for the restaurant industry, the end of the tunnel is nowhere in sight.


text: Media CenterEvents ICANN Regional Meeting 4/1/2008 Page Content The United Arab Emirates is hosting the regional meeting for the Internet Corporation for Assigned Names and Numbers (ICANN) in the period from 1 to 3 April 2008. The meeting will address a number of major issues, namely: Internationalized Domain Names IDN Challenges and opportunities in registry/ registrar business Current developments in the generic Top-Level Domain Space The meeting will furnish all stakeholders with an opportunity to exchange expertise and opinions. It constitutes a platform for the Arab region to get introduced to the latest related technologies and developments. Meetings will also tackle the challenges and difficulties facing Arabic Internet users. The Egyptian delegation will submit a paper on the second day of the meeting on Top Domain Names in local languages. For more info, please visit:http://public.icann.org/dubai08

subset: commoncrawl


text: HomeArtist Of The MonthArtist Of The Month – Armin Van Buuren (December 2018) Rajrishi Murthi December 2, 2018 Artist Of The Month, FeaturedBox This man needs no introduction. And rightfully so, this legendary figure continues to command his position as one of trance and electronic dance music's biggest and most renowned names over the past two decades. As you might have guessed by now, we're indeed talking about the one and only, Armin Van Buuren. What has turned out to be yet another massive year for electronic music throughout the globe, we're all set to bring this year's chapter to a close by putting the spotlight on unarguably, one of the greatest names ever to be a part of the scene. As the Dutch heavyweight gets ready to make his way back to the Indian sub-continent later this month for his headlining performance at Sunburn Festival, please welcome our Artist Of The Month for December 2018, Armin Van Buuren. While many recognise him for his iconic ASOT (A State Of Trance) radio show and some of his biggest global hits in recent times including 'Another You', 'I Need You', 'Sunny Days', 'Sex, Love & Water', 'Therapy', 'Blah Blah Blah' amongst others, Armin Van Buuren's beginnings date back to the late 90s when he kicked off his illustrious career with the release of his debut single, 'Push' in 1996. What followed soon after was the release of his breakout track, 'Blue Fear' in 1997. As Armin continued to push his way through during the earlier part of his career, listeners were treated to cuts like 'Communication' which also appeared on his 2003-released debut studio album '76'. In 1999 and 2000, Armin Van Buuren dished out several releases including cuts like 'Free' and 'Touch Me' under his Gimmick and Rising Star alias. With the debut of his Alibi and Major League projects with Tiesto in 2000 which saw them put out tracks like 'Eternity' and 'Wonder Where You Are?', he also shared his highly acclaimed collaboration with Ferry Corsten, 'Exhale' and debuted his alias Gaia with the release of '4 Elements', which now consists of two members, including Armin and Benno de Goeij. Come 2001, the Dutch act finally brought his now iconic radio show, A State Of Trance to life in March. Later that year, he also debuted the track 'The Sound Of Goodbye' as Perpetuous Dreamer. Armin Van Buuren went on release his debut album, '76' in 2003 which boasted of iconic cuts like 'Yet Another Day', 'Burned with Desire', amongst other tracks. During their period, he also introduced his hit remix of 'The Longest Day'. During that year, Armin also launched his very own label, Armada Music along with Maykel Piron and David Lewis. In 2005, Armin came out all guns blazing as he not only completed 200 episodes of A State Of Trance, but also went on to release his second LP, 'Shivers' which featured collaborations with Nadia Ali, Justine Suissa, Gabriel & Dresden amongst others. With releases like 'Control Freak', 'Love You More', 'Rush Hour' and many more which paved his way forward, 2007 witnessed Armin being crowned as the #1 DJ in the world as part of the DJ Mag Top 100 DJs rankings. Next came his third album, 'Imagine' which spawned tracks like 'Going Wrong', 'In and Out of Love' and 'Unforgivable'. With his ever growing success which continued to scale new heights, he went on to be crowned as the World's #1 DJ for the third consecutive year in 2009. 2010 brought upon the release of his highly acclaimed album, 'Mirage' which was also followed by the Armin Only World Tour. As his globally loved radio show, A State Of Trance crossed the mammoth landmark of completing 500 episodes in 2011, Armin delivered five exceptional live shows across the globe in celebration of this huge feat. While 2012 saw him reclaim his #1 spot on the DJ Mag Top 100 DJs rankings, listeners were also graced by several other memorable releases like 'Youtopia' and 'We Are Here To Make Some Noise'. In 2013, the iconic act served up his fifth studio album, 'Intense' which also housed some of his biggest hits till date including the Grammy nominated single, 'This Is What It Feels Like', 'Waiting For The Night', 'Beautiful Life' amongst others. With the success of other tracks like 'D# Fat' with W&W, 'Save My Night' and 'Ping Pong', Armin continued to dominate festival stages around the globe. One of dance music's busiest DJs, Armin Van Buuren headlines more than 150 shows in a calendar year at some of the world's biggest festivals and clubs. With appearances at festivals like Tomorrowland, Ultra Music Festival, Electric Daisy Carnival and many more, the Armada bossman continues to enthrall audiences with his energetic performances till date. And in case you needed some more proof regarding his love for music, then it certainly needs to be known that Armin still serves as A&R of Armada's very own sub imprints, Armind, Who's Afraid Of 138?! , and A State of Trance. In 2015, Armin marked the arrival of his sixth studio album 'Embrace' which was led by the release of his first single, 'Another You' with Mr Probz. Later that year, he also unveiled his mighty remix of Game of Thrones' official theme composed by Ramin Djawadi, alongside the release of other collaborations with the likes of Hardwell, Cosmic Gate, W&W, Jean Michel Jarre and many more. What followed next was the return of his iconic Armin Only world tour in support of his latest LP which took place throughout the globe throughout 2015 and 2016. Next came the unveil of his mammoth 'Old Skool' EP which housed in one of 2016's biggest tunes, 'Dominator' with Human Resource alongside other tracks like 'The Ultimate Seduction' and 'Pull Over'. Eventually, Armin brought 2016 to a fantastic close with the release of the official ASOT 800 anthem, 'I Live For That Energy' and 'Great Spirit' alongside Vini Vici and The Hilight Tribe. While the superstar act may have faced flak in recent years over his shift from trance to exploring more festival ready sounds, Armin's legacy has remained intact with the consistent onslaught of releases and live performances around the globe. In 2017, Armin Van Buuren took over the Amsterdam Arena to host his The Best Of Armin Only show to commemorate the 20th anniversary of his career which was also accompanied by the release of a compilation album. Furthermore, with releases like 'I Need You', 'Sunny Days', 'You Are' with Sunnery James & Ryan Marciano throughout 2017, he managed to secure yet another successful year before stepping into 2018. And now with 2018 set to come to a monumental close for Armin Van Buuren, the legendary name can certainly tick off yet another successful year which saw him release his fan favourite 'Blah Blah Blah' EP led by its titular cut, alongside other tracks like 'The Last Dancer', 'Just As You Are' and 'Popcorn'. With other brilliant releases like 'Sex, Love & Water', 'Our Origin' with Shapov, 'Wild Wild Son' and his most recent collaboration, 'Ready To Rave' with W&W, the Dutch act has made sure to keep the music coming for his fans around the globe. Additionally, he also delivered a record breaking seven hour long headlining performance at Untold Festival in Romania earlier this year. With all eyes on AVB himself as he gets ready to shut down 2018 with some of his highly anticipated end of the year performances, we can't wait to see what Armin has in store for us in 2019.


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On January 9, 2018, Spectra7 Microsystems Inc. closed a bought deal offering of 15,315 units at a price of $1,000 per unit for aggregate gross proceeds of $15.3 million, which included the partial exercise of the over-allotment option granted to the underwriter for 315 units. Each unit consisted of one 7.0% senior unsecured convertible debenture and 1,425 common share purchase warrants. The principal amount of each convertible debenture is convertible into common shares at a conversion price of $0.35 per common share at any time prior to January 9, 2021. Each warrant entitles the holder to acquire one common share at a price of $0.50 until January 9, 2021. The offering was underwritten by Canaccord Genuity Corp.
Cassels acted for Cannaccord with a deal team that included Andrea FitzGerald, Alex Pizale, Matt Gold and Xi Chen (Securities), Jim Morand (Tax) and Jason Arbuck (Financial Services).


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Julia Skinner is a doctoral student at Florida State University's School of Library & Information Studies. Her research interests include social media, library history, and services for sexual assault survivors. She is the instructor for two classes coming up with Library Juice Academy in August: Social Media for Libraries, and The Librarian as Scholar: Taking Part in Scholarly Communication. She agreed to do an interview here in order to give people a better sense of what will be covered in these classes and how they might benefit from them, and a little bit more about her and her varied interests.
Hi Julia! Thanks for agreeing to do this interview.
I think I'd like to start by asking you to talk a bit about your background in terms of your qualifications and interest in teaching these two classes.
Both of these are areas I'm really passionate about. I'm interested in social media both as a user and a researcher. My social media presence (@BookishJulia) is a hybrid of personal and professional, which is an approach I really like for my own work. I have also done some social media managing work, and am in the process of drafting a social media policy and editing it as I receive feedback. That process is really important, and it's something I can't wait to share with students. With that policy, you have the groundwork laid for what your institution's social media presence will look like, and it helps hold people accountable for what they can post and how often, as well as how they interact with others in that space. I also do research on social media as it is used during political protests and during natural disasters. I'm really interested in how social media can be a force for good that connects us to other people and that can be harnessed to share resources, which is one of the reasons why I think it's such a great tool for libraries looking to connect with patrons.
For scholarly communication, I have experience in a variety of areas. I was co-editor of a student-run journal (B-Sides) during my MLS, I have published a number of articles in academic journals, have interned at another journal (JELIS), and have served as a reviewer for a handful of journals as well. I am excited to teach this class because feel like the writing and publishing process can be intimidating for people who aren't in academia, or who are new to it. A lot of students don't get training in scholarly communications in their programs, so they aren't really sure how to approach that process when they want or need to begin publishing their work. I would love to see more practitioners share their research and ideas in the literature, because there is so much incredible work taking place in our field, but not all of it is getting the attention it deserves.
Cool. Regarding scholarly communication, it seems like knowing more about it could be useful both as a scholar and as a librarian in an academic setting. I think that that class will be most attractive to people who are interested in both sides of it.
Before we go further, I think I'd like to ask you to describe both of the classes – say what they cover and what you expect participants to take away from it at the end or be able to do.
I agree! I think learning about scholarly communication is something that is valuable for just about anyone. I designed both classes to give students activities and knowledge that they can immediately take out in the world and start using.
For the scholarly communication course, we'll be covering the manuscript submission process, the review and revision process, and writing conference abstracts. Since the course is short, we will be using existing manuscripts (either one the student is already working on or one that I provide them) so we can spend more time discussing the submission and review process. Hopefully by the end of the course, they will understand what each step entails, will have a list of journals to submit their work to (along with feedback to polish it up before it goes out the door), and will have a conference abstract prepared. I also want them to feel more comfortable with the overall process to the extent that when they get back journal reviews they don't feel intimidated or disheartened, so that all of us in LIS can learn from their work.
For the social media course, we'll be covering how to use social media effectively, and we'll also be writing a social media policy and thinking about how to implement the policy once it's written. I encourage a lot of interaction in the other course too, but since this one is about technology that is inherently social, I want everyone to be sharing what they're thinking and learning from each other. Some of the folks in the class are probably very comfortable with these technologies already, but that way those who are less familiar can get used to being social and engaged in an online space. I want the people who take this course to walk away with an understanding of what social media is and why it's important, and what an effective social media presence would look like for them and their organizations. I also want to underscore the importance of having agreed-upon guidelines to drive their social media presence, and have them walk away with a set of guidelines that they think will work for their situation, that include the feedback of everyone else in the class. The last week of class will be one where we think about implementing a policy and evaluating its effectiveness, which I hope will give them some ideas for how to broach the subject with peers and supervisors, and create a lasting and evolving social media presence.
Sounds very good. Let's talk a little bit about the scholarly communication course. I want to make sure it's clear that this course is about the process of writing, revising, submitting, etc., and not about the research process, which is a related but different set of skills that can vary according to what kind of paper it is we're talking about. We have another class on research methods for internal research within a library, but we don't have a class on research methods as it might be applied to research done for publication. For the purpose of your class, we are going to assume that participants have that covered in some other way, or that they are writing a kind of paper where it is not an issue. Does that sound correct to you, and do you have anything to add to that?
You're absolutely right. This course deals with publishing your work after it's completed. Research design is a completely different thing with its own set of considerations. I am assuming that when participants come into the class, that they have already had some exposure to research for publication, even if they don't feel like they're experts at it. If someone doesn't have that experience, I think they would still get a lot out of the class that they could apply after they learned about methods and started doing their own work, but having some knowledge of research methods (or some experience conducting research) will make it easier.
Yeah. Though with more humanities-based research that element isn't there so much. And of course, for librarians who aren't trying to get some research published but who just want to learn more about the process to enrich their professional knowledge as academic librarians, I think they also wouldn't have so much need to learn research design. But we do have a class that gets into it, with a focus on studying users or processes at your own institution.
Staying with the scholarly communication course for a bit, I'd like to ask you about your own experience writing for publication and going through the process. Have you been surprised by any aspects of it?
I have had one thing that has especially surprised me. One of the things that always sticks out to me is the things editors and reviewers focus on that I've done well or need to improve, because usually they are different than what I would target as the strongest and weakest points of a paper. I always end up learning a lot from the different perspectives that they bring to my work, and I think it highlights the importance of the review process because you get to understand your writing from so many angles.
That's cool. So it sounds like going through the process can be very educational, and one thing this course can do is to provide encouragement and give people a good start.
For sure! I think the process is a lot of fun, and once you have a sense of what to expect it makes it easier to be a part of.
I'd like to turn to the social media class. There are a lot of classes on social media out there. It seems like something that is different about this one is the emphasis on establishing guidelines and then evaluating their effectiveness. I like that – it seems very practical and nicely directed toward insitutional settings. I think at this point we can assume that everybody who has the responsibility of managing social media for a library has a personal social media presence. I am wondering what besides having guidelines is going to be different for an institutional social media presence, as opposed to a personal one?
Well, one thing that might be different is how widely you want to share your posts. I know on my Facebook account, I have limited my posts so they are only visible to friends.However, with your institutional account, you want your posts to be publicly available. One thing I bring up in the class is the overlap between personal and institutional approaches, which I think is especially useful since so many people (like me) have hybrid personal-professional accounts, and their experiences with those accounts can give them some ideas of what has and hasn't been working to increase engagement and visibility.
I also share different content in different spaces, which is something that might be appropriate for an institution's social media presence, but the type of content being shared is very different. On all my accounts, I share things of interest to personal and professional contexts, but I'm basically doing it to curate my own presence in an online space. For an institution, the only goal may not be to curate an image, but may also be to increase awareness or bring in more patrons, so the type of posting being done will be different.
Makes sense. Another thing I'm wondering about is how in an institutional setting there may be other people involved, at least in the sense that guidelines are going to have to be approved, and evaluation of how it is going may not be done only by you. Does that bring in different kinds of issues that you plan to get into?
Yes–one thing I stress is that it's a collaborative process, and that the policy we draft during the class can be a helpful jumping off point for everyone who's involved in making that decision. Having something specific in writing gives people a chance to point to things they like and things they want to change, so the final document ends up being something everyone agrees on.
You also have the potential to include multiple people in managing the accounts as well. I urge everyone to clearly outline who does what (e.g. posting), and who replaces or fills in for those people when they aren't available.
Good. Something that your class makes me think about is how by this point, it is commonplace for library institutions to have a social media presence, and it also seems like in most cases their presence is not doing much for them. Would you agree with that? And if so, do you think that it is clear what they should be doing differently? I am wondering what the state of knowledge is about social media for institutions, especially library institutions. I am thinking that a lot of people right now feel competent to jump in, but maybe there is a lot they should know about making their institutional social media presence actually work. What do you think about that?
I would definitely agree. I've learned a lot about social media and libraries by looking at examples of what libraries are doing, and there are huge differences in engagement, frequency of posting, types of posts, etc. It's not always clear, but even if it's not I think there's a lot of value in trying different approaches until you find something that sticks. In the class, I will be encouraging students to think about their own institution and use that knowledge to guide what they're doing. What works in one place won't work in another, and your target audience might be using different platforms or respond to different types of posts than the target audience of another library.
I would agree that a lot of people feel competent to jump in, particularly since creating an account is so easy. One thing that's a potential concern is spreading yourself too thin–you can easily create accounts, but will you have the time to continuously fill them with quality content? I think having at least some guidelines in place, if not a full-fledged policy, will help put everyone on the same page and help clarify what will be posted and how interactions will look before there's the opportunity for confusion.
One thing about social media is that it's a place where mistakes can be amplified quickly since content can be re-shared and because users have the ability to interact directly with you in a public setting. Hopefully this wouldn't happen, but an example would be if someone posted a complaint about the library and the social media manager ignored it or got defensive rather than apologizing and offering solutions. That person might go onto their account and complain to all their followers about the library, which then has a negative impact on many more people's perception of the library. I don't think managing social media needs to be intimidating, but I do think it's a matter of being clear about what you want from social media, experimenting until you find effective ways to get that, and being engaged with users.
Sounds good. Sounds like the class will involve a lot of good discussion and sharing of experiences.
I hope it will!
I'd like to finish up by asking you about some of your other interests, which I know are varied. First, you're working on a Ph.D. Do you have a dissertation topic? How is that going?
I'm just about to start my third year of my program, so I'm finished with my coursework, but am preparing for my prelim exam (my qualifying exam) before I start on my dissertation. With any luck, I'll get that out of the way later this year or early next year, and then I'll start doing that research. My dissertation topic is going to be one librarian who worked in the Harlem Public Library in the 1930s. She did a lot of cool stuff to make the library a community center, and from what I understand there are a lot of records on her work still around. Depending on what I find, I would also like to bring in some LIS theory to help me frame the stories of her and her patrons. This time period is interesting because it's a time of uncertainty and change, which is something that is a thread through all of my research areas, and I'm looking forward to seeing how that plays out in a library.
Well that sounds very cool. Can I link out to your CV from here? Given the breadth of your interests, I have to ask as a final question – If you could teach anything for Library Juice Academy, what would it be?
Thank you! You most certainly can link to my CV. Actually, the two courses I'm teaching are pretty ideal–they're both areas I have experience in and am passionate about, so I love the idea of getting to share what I know and learn alongside students as we go through the material. If I were going to teach a course in addition to them, it might be fun to do a research methods course that focuses on approaches that might not be covered in other methods courses, particularly since I'm doing research using a few different methods in a few different areas.
Sounds great! Thanks very much for this interview, and best of luck with your classes next month.
Thank you! I'm looking forward to them!


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I have not been to a Faith on Tap in quite awhile (which makes sense since I am no longer of the age group for this event), but hopefully I will stop by sometime because they were always interesting and fun.
"Being A Young Catholic in the 21st Century"
All Young Adults
You are invited to join us on
Monday, April 22 for our Next Faith on Tap
us on Monday, April 22, as Paul Morisi discusses the challenges that
face young adults in the church. Why do young adults leave the Catholic
Church ... and why do others stay? What does the Church offer young
adults in the 21st Century? Come out and join us as we discuss these
Faith on Tap will be held at the Wantagh Inn
3264 Railroad Avenue
Across from the RR Station
Doors will open at7:00
Come out early ~ have dinner ~ meet new friends ~
re-connect with old friends
Talk begins at 7:30


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Twice this year, I have stumbled upon co-workers and ex-coworker's living spaces on local blogs. Both are ENVY worthy. Both make me think that the internet is a strange bird. Ten years ago, I would never have seen inside the house of a casual acquaintance- and now, welcome inside, internet!
And – how much do I LOVE that side shelf in the master bedroom of Mike and Anne? A whole, whole lot.


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Town of Danville residents and business owners, join MCE staff at your Town Council Meeting for an insightful glimpse into Community Choice Aggregation (CCA). This local public utility model is making waves throughout the State of California, allowing cities, towns, counties and communities, to greatly reduce their energy related carbon footprints. Come learn how this is possible! Have your questions and comments ready.
For Board and Committee meetings, please click here.
You can find us at any of the community events or meetings listed below. If you need more information, feel free to contact us at email@example.com.
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text: We are selling a condo unit "One Central Park", located at Eastwood City - a Megaworld corporation developer. Our condo unit is for sale at pre-selling rate, this condo has never been use. The room is perfect for singles, couples, or small families because of the comfortable unit cuts of 46.5 square meters. The rooms … Continue reading For sale condo unit at pre-selling rate, One Central Park – Eastwood City


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100 Years on from the Suffragette movement.
The Birmingham Jazz Legends Festival 2018 has been announced and will mark Birmingham Jazz's contribution to the centenary celebrations of female emancipation.
The Festival features the front line of Jazz Suffragettes, today's players demonstrating their own emotional journeys and those of the past – celebrating jazz women. Each element of the festival will bring a woman performing jazz into focus.
Over the six days there will be fourteen gigs, featuring a variety of national bands and legendary names and the very best of local jazz talent.
Tickets are available from www.birminghamjazz.co.uk (booking fee applies) or you can call Phil Rose on 07887 526435 to arrange payment by cheque or cash (no fees).


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Ahead of our trip to face Manchester United at Old Trafford, we sat down with a man who has enjoyed his fair share of derby experiences - David Silva
The midfield maestro has featured in sixteen matches against United since arriving at the club in 2010, playing a major part in many of our derby day delights over that period.
READ: United v City: Boss reveals De Bruyne latest
WATCH: How the last Manchester Derby was won
From a goal and incredible assist in the 6-1 away win back in 2011 through to the early opening strike in November, the Spaniard has regularly been a key cog in the well-oiled City machine.
Watch the video above to recall some of David's favourite derby moments…


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There remains uncertainty and some controversy about the percentages and types of cells in human sensory nerve ganglia that harbor latent herpes simplex virus 1 (HSV-1) and varicella-zoster virus (VZV) DNA. We developed and validated laser-capture microdissection and real-time PCR (LCM/PCR) assays for the presence and copy numbers of HSV-1 gG and VZV gene 62 sequences in single cells recovered from sections of human trigeminal ganglia (TG) obtained at autopsy. Among 970 individual sensory neurons from five subjects, 2.0 to 10.5% were positive for HSV-1 DNA, with a median of 11.3 copies/positive cell, compared with 0.2 to 1.5% of neurons found to be positive by in situ hybridization (ISH) for HSV-1 latency-associated transcripts (LAT), the classical surrogate marker for HSV latency. This indicates a more pervasive latent HSV-1 infection of human TG neurons than originally thought. Combined ISH/LCM/PCR assays revealed that the majority of the latently infected neurons do not accumulate LAT to detectable levels. We detected VZV DNA in 1.0 to 6.9% of individual neurons from 10 subjects. Of the total 1,722 neurons tested, 4.1% were VZV DNA positive, with a median of 6.9 viral genomes/positive cell. After removal by LCM of all visible neurons on a slide, all surrounding nonneuronal cells were harvested and assayed: 21 copies of HSV-1 DNA were detected in ∼5,200 nonneuronal cells, while nine VZV genomes were detected in ∼14,200 nonneuronal cells. These data indicate that both HSV-1 and VZV DNAs persist in human TG primarily, if not exclusively, in a moderate percentage of neuronal cells.
Herpes simplex virus type 1 (HSV-1) and varicella-zoster virus (VZV) establish lifelong latent infections in human sensory ganglia, processes that have been investigated extensively but are still not fully elucidated. It is well established that during latency, infectious HSV-1 and VZV particles are not produced (40, 41, 56), but small subsets of their genes are expressed (11, 12, 13, 21, 25, 27, 31, 35, 44, 52). These latent viruses, however, are subject to single or multiple rounds of reactivation and can result in recrudescent disease (7, 59). For HSV, gene expression in latency is extremely restricted in that only the latency-associated transcripts (LAT) accumulate to levels high enough to be detected by in situ hybridization (ISH) (4, 11, 13, 44, 49, 50, 52). Although there is no confirmation that LAT encodes a protein that regulates HSV latency and reactivation, it has been documented that the 5′ portion of the LAT gene facilitates the efficient establishment and reactivation from latency in experimentally infected animals (3, 10, 16, 23, 38, 48, 54, 55).
The discovery and detection of LAT by ISH enabled an indirect estimate of the percentages of animal and human sensory ganglion neurons that are latently infected. ISH studies in our laboratory, for example, showed that LAT can be detected in 0.2 to 4.3% of the neurons in human trigeminal ganglia (TG) (11), a range similar to that reported for experimentally infected mice, rabbits, and guinea pigs (4, 22, 44). Because the in situ detection of LAT is merely a surrogate marker for HSV latency, it is possible that many more neurons are latently infected but their LAT expression or accumulation is too low to be detected by this technique. In accord with this possibility, a variety of tools have been used to better estimate the numbers of HSV-1 DNA-containing neurons in experimental animals, including in situ PCR (27, 33, 34, 42, 43), contextual analysis (45, 47), and laser-capture microdissection (LCM) (6). These studies revealed that, in mice and rats, neurons that are LAT positive by ISH represent but a fraction of those harboring HSV-1 DNA (6, 33, 42, 45).
In the past several years, using real-time DNA PCR assays, the latent HSV-1 DNA load in human TG was estimated to be hundreds to thousands of copies/μg of total ganglion DNA (9, 39), suggesting that a higher proportion of cells might be latently infected than are identified by ISH. The actual percentages of neurons that harbor latent HSV-1 DNA and the ranges of viral genome copy numbers in individual neurons, however, were addressed more directly only recently by PCR of dissociated human TG cells (5).
As complex as the analysis of HSV latency in human ganglia has been, studies of the cellular distribution of latent VZV DNA in human ganglia have been even more so, despite numerous attempts using ISH (12, 21, 25, 27, 31), in situ PCR (17, 27), immunohistochemical staining (8, 26, 32, 36), and PCR of dissociated ganglion cells (28, 30). There had long remained controversy, for example, as to which cells primarily harbor latent VZV. Our laboratory reported many years ago, for example, that by ISH, VZV transcripts were most evident in nonneuronal, satellite cells (12). Yet, aggregate data from many other laboratories have since built a compelling case that the primary cellular locus of VZV latency is the sensory neuron (17, 21, 25, 27, 28, 30), although the percentages of neurons reported to be VZV positive in these prior studies have ranged very widely, from 0 to 1.5% of cells in some studies to 20 to 30% of cells in others, depending on the methodologies employed (25, 30, 31).
To further refine estimates of the cellular distribution of latent HSV-1 infection and to clarify the cellular locus and distribution of latent VZV infection in human TG, we have been exploring the use of LCM, a method for visualizing and procuring single cells from conventional histological sections for analysis. LCM utilizes an infrared laser integrated into a microscope to activate a small area of a thermoplastic membrane attached to a transparent cap that is placed directly on the surface of the section mounted on a glass slide. When heated by the laser pulse, the membrane is activated, causing it to bind the targeted cells below it (Fig. 1) so that their DNA, RNA, or protein contents can be extracted for analyses (19). To date, LCM has been used primarily for analyses of gene expression profiles in cancer cells (14, 18, 20). By using LCM followed by real-time PCR assays (LCM/PCR) for HSV-1 and VZV DNA in human TG and comparing the results with those of ISH for LAT, we have documented the persistence of HSV-1 in a percentage of human neurons, of which only a subset accumulates detectable LAT. Moreover, we detect VZV DNA in a moderate percentage of neurons, with little or no viral DNA in nonneuronal cells.
TG and serum samples.TG were collected at autopsy at a postmortem interval of less than 24 h from 11 subjects, whose salient historical and clinical features are summarized in Table 1, and were immediately fixed in 10% formalin in phosphate-buffered saline at room temperature overnight and then embedded in paraffin. One TG of each subject from whom serum was not available for antibody assays was snap-frozen on dry ice and stored at −80°C for later extraction of total DNA and determination of the status of HSV-1 and VZV infection. From 7 of the 11 subjects postmortem serum samples were collected and frozen at −20°C for later serological testing.
Serological analyses.Titers of type-specific human serum antibodies against HSV-1 were determined by means of Western blot analysis in the laboratory of Rhoda Ashley at the University of Washington in Seattle, as described previously (1). Titers of serum antibodies to VZV were determined in the Department of Clinical Pathology, Clinical Center, National Institutes of Health, using the Captia VZV immunoglobulin G kit (Trinity Biotech USA, New York), according to the manufacturer's instructions. With these assays we determined that all subjects tested were seropositive for both VZV and HSV-1, with the exception of subjects 701 and 710, who lacked detectable circulating antibodies to HSV-1.
HSV-2 LAT transgenic mice.TG from the HSV-2 LAT transgenic mice we engineered and have described previously (57) were used to help define the efficiency of LCM in recovering individual neurons and the overall sensitivity of the assay. For these experiments, TG harvested from transgenic line 5238, which bears ∼40 copies of the transgene/cell, were processed as described above for human TG.
In situ hybridization.RNA sequences antisense to LAT, corresponding to nucleotides 119,505 to 120,669 of the HSV-1 genome, were transcribed in vitro from plasmid pG.1LAT using a SP6/T7 transcription kit (Roche Molecular Biochemicals), according to the manufacturer's instructions. This RNA product was then fragmented by partial alkaline hydrolysis. Briefly, a 100-μl reaction mixture containing 48 μl of RNA, 10 mM dithiothreitol, 60 mM Na2CO3, and 40 mM NaHCO3 was incubated at 64°C for 50 min. The reaction mixture was then chilled on ice, 5 μl of 10% glacial acetic acid and 10 μl of 4 M sodium acetate, pH 5.2, were added to it and mixed, 300 μl of cold (−20°C) ethanol was then added, and the mixture was incubated at −20°C overnight. After centrifugation at 18,000 × g, the pellet was washed with 70% ethanol and resuspended in 40 μl of RNase-free water.
To prepare the biotin-labeled RNA probe, 5 μg of RNA in 20 μl Tris-EDTA buffer was mixed with 20 μl of 1 μg/μl PHOTOPROBE biotin reagent (Vector Laboratories, Burlingame, Calif.), according to the manufacturer's instructions for the thermal coupling method. Briefly, the reaction mixture was covered with 30 μl of mineral oil and incubated at 95°C for 30 min. The aqueous phase was taken and mixed with 40 μl RNase-free water, 80 μl of 100 mM Tris-HCl pH 9.5, and extracted twice with 160 μl of 2-butanol to remove the unincorporated biotin. The aqueous phase recovered from this extraction procedure was supplemented with 10 μl of 10 M ammonium acetate, 2 μl of 1 M MgCl2, and 125 μl of cold (−20°C) ethanol and then incubated at −20°C for 15 min. After centrifugation at 18,000 × g for 15 min, the pellet was washed with 70% ethanol and resuspended in 40 μl Tris-EDTA buffer. This probe was stored at −80°C in aliquots sufficient for individual reactions.
ISH was then performed, as described previously (11), with some modifications required to adapt the procedure for the use of the biotinylated probe and so that the sections could be later subjected to LCM. Briefly, TG sections were deparaffinized twice in serial xylene baths for 5 min each; incubations were carried out at room temperature unless otherwise specified, followed by rehydration sequentially in 100%, 100%, 95%, 70%, and 50% ethanol for 2 min each and 1 min twice in water. Slides were then transferred to 0.2 N HCl for 20 min, rinsed in water, treated with prewarmed 25 μg/ml proteinase K, 10 mM Tris-HCl, pH 7.4, 2 mM CaCl2 at 37°C for 15 min, and finally rinsed in water. Acetylation was performed by incubating the slides in freshly prepared 0.25% (vol/vol) acetic anhydride/1.33% (vol/vol) triethanolamine at pH 8.0 for 10 min and finally rinsing the slides in water. Excess water was wiped from the slides, taking care to not touch the tissue sections themselves. Then, 30 μl of prehybridization solution (50% formamide, 2× SSC [1× SSC is 0.15 M NaCl plus 0.015 M sodium citrate], 50 mM Tris, 1 mM EDTA, 0.02% Ficoll, 0.02% polyvinylpyrrolidone, 500 μg/ml tRNA, 500 μg/ml bovine serum albumin) was added onto each section. One minute later the liquid was drained, another 20 μl of the prehybridization solution was added onto each section, and then the section was covered with a glass coverslip. After incubation at 42°C for 2 h, the coverslip was removed and 30 μl of hybridization solution (50% formamide, 20% dextran sulfate, 1 mM EDTA, 10 mM dithiothreitol, 300 mM NaCl, 50 mM Tris-HCl, pH 7.4, 0.02% Ficoll, 0.02% polyvinylpyrrolidone, 500 μg/ml tRNA, 500 μg/ml bovine serum albumin) with 500 ng/ml of heat-denatured (65°C for 20 min) probe was added. New coverslips were applied, the edges were sealed with rubber cement, and slides were incubated at 55°C overnight. The next day, after the sealant and coverslips were carefully removed, slides were washed twice with 2× SSC for 10 min each at room temperature, twice at 55°C for 20 min each with buffer containing 2× SSC, 1 mM EDTA, and 0.1% Triton X-100, and once more for 30 min at 55°C with buffer containing 0.1× SSC, 1 mM EDTA, and 0.1% Triton X-100. To remove any unhybridized RNA probe, sections were treated with an RNase mixture containing 40 μg/ml RNase A, 10 U/ml RNase T1, 10 mM Tris-HCl, pH 7.4, and 300 mM NaCl, incubated at 37°C for 40 min, and finally washed twice for 20 min each with 2× SSC at 55°C. To detect the hybridized probe, slides were incubated in a blocking solution of 1× casein in 100 mM Tris-HCl (pH 7.5), 150 mM NaCl, and 0.1% Tween 20 (TBST) for 40 min. Horseradish peroxidase-streptavidin (Vector Laboratories) was added to the blocking solution to yield a final concentration of 4 μg/ml, and slides were incubated for another 30 min. After three washes in TBST, the slides were developed using a TMB substrate kit for horseradish peroxidase (Vector Laboratories) for 5 min and then rinsed in water. Slides were dehydrated in graded 70%, 95%, 100%, and 100% ethanol for 1 min each and 1 min twice in xylene, and then air dried.
LCM.Serial 5-μm sections from different anatomical regions of each TG were cut from each tissue block and mounted on plain glass slides. In order to better represent the neurons in whole ganglia with limited numbers of cells that can be processed by LCM/PCR, cells from the first, middle, and last 10 slides of a ganglion were selected for LCM. The tissue sections were deparaffinized twice in xylene at room temperature for 5 min each and then air dried. LCM was performed using a PixCell II (Arcturus Engineering, Inc., Mountain View, Calif.). The laser beam was set to be 7.5 μm in diameter, with a power of 25 to 35 mW and for 30 to 40 ms in duration. These optimal settings were selected for capturing single neurons from human TG without apparent contamination with adjacent cells. After placing CapSure LCM caps (Arcturus Engineering, Inc.) on top of each tissue section, single neurons with visible nuclei were chosen under the microscope and a laser pulse was fired to capture that neuron onto the cap. Satellite cells, hundreds on each cap, were harvested only after all of the neurons in the desired area were removed.
To procure LAT-positive and LAT-negative neurons separately, sections were first subjected to ISH for HSV-1 LAT, as described in the previous section. The in situ-stained cells were then visualized and photomicrographed while the sections were still covered with xylene. After each section was air dried, LAT-positive and -negative cells were identified with the guidance of the photographs and captured using LCM.
DNA extraction of LCM samples.Caps with captured cells were inserted into 0.5-ml GeneAmp tubes (Applied Biosystems) containing 14 or 50 μl of DNA extraction buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA, 1% Tween 20, and freshly added proteinase K to yield 0.04%) for single neurons or multiple satellite cells, respectively. At 37°C overnight, the tubes were incubated upside down so that all captured cells were covered with the extraction buffer. After 1 min of centrifugation at 18,000 × g, the caps were removed and the tubes were sealed and heated at 95°C for 8 min to inactivate any remaining proteinase K. From each sample, 12 μl/reaction mixture was subjected to real-time PCR specific for HSV-1 glycoprotein G (gG) or VZV immediate-early gene 62, as described previously (39). The sequences of the primers and probes used are shown in Table 2. Each 25 μl of the PCR mixture contained 12.5 μl of 2× Universal TaqMan PCR master mix (Applied Biosystems), forward and reverse primers at a final concentration of 1.0 μM each, 0.2 μM of probe, and 12 μl of DNA extract from the LCM sample. In every PCR assay performed, four no-template control reactions and a standard panel of DNA copy-number reference samples were run. Each reference panel consisted of six dilutions of the pcDNA3.gG1 plasmid DNA for quantifying HSV-1 DNA, or pCMV62 DNA for VZV DNA, at 1.5, 5, 50, 500, 5,000, or 50,000 copies/reaction mixture. All standards were run in duplicate. All reactions were carried out in an ABI PRISM 7700 sequence detector (Applied Biosystems, Foster City, CA) with optimized thermal cycle conditions: 50°C for 2 min, 95°C for 10 min, and 40 cycles at 95°C for 20 seconds and 60°C for 1 min. By comparing the threshold cycle (Ct) of each sample to the Cts from the standard reference panel, the copy number of each experimental reaction was then determined. To estimate the cellular genome content of LCM samples of satellite cells, 24 μl out of 50 μl DNA extract from each cap with satellite cells was subjected to PCR in duplicate with primers and probes specific for human albumin, with human genomic DNA (Applied Biosystems) as copy number reference.
Statistical analyses.Analysis of variance and linear regression analyses were performed with the computer software package JMP version 3.0 (SAS Institute, Cary, NC).
LCM/PCR detection of viral DNA in single cells is sensitive, specific, and efficient.Extensive developmental work was undertaken to define the operational characteristics and limits of the LCM/PCR assays for detecting viral DNA sequences in single human neurons. We began by determining the optimal LCM pulse characteristics. In this regard, we determined microscopically that the diameter of human TG neurons in fixed tissue sections ranged from 25 to 60 μm. When the energy and diameter of the laser beam were set for each desired TG neuron, as specified in Materials and Methods, it activated an area of the thermal membrane slightly smaller than the diameter of that cell, such that the content of the targeted single neuron was captured while all surrounding satellite and other nonneuronal cells were left attached to the slide (Fig. 2).
Even though we performed LCM only on cells whose nuclei were visible in the section, because of both the depth of the tissue sections and potential inefficiencies in extraction and detection, we expected that the DNA recovered from each cell might represent only a fraction of its entire content. Thus, we set out to estimate the efficiency of cellular DNA recovery and detection by LCM/PCR. To this end, we first tested the LCM/PCR efficiency of single neurons recovered from TG sections of heterozygous HSV-2 LAT transgenic mice that we had engineered and estimated previously by Southern hybridization to harbor about 40 copies of the LAT transgene/cell (57) instead of directly testing single neurons from human TG, because the copy number of the cellular gene is too low to be detected. By LCM/PCR an average of 37.4 ± 3.6 (standard error of the mean) copies of the HSV-2 LAT genomic sequence were detected from DNA extracted from single TG neurons of the transgenic mice. The result was not affected by adding 10 ng of human genomic DNA or total RNA isolated from the transgenic mouse liver in an amount equivalent to about 5,000 copies of LAT RNA. Thus, by this method, it appeared that the combined efficiency of LCM/PCR in recovering DNA from a single neuron was roughly 93.4 ± 9%.
The combined efficiency of LCM/PCR of sensory neurons from human TG was also examined by subjecting DNA extracted from LCM caps containing known numbers (50 to 250) of neurons to PCR with primers and a probe for the human albumin gene (there are two copies of the gene per cell). Based on the numbers of albumin gene copies recovered, we estimated an efficiency of 64 ± 15% for LCM/PCR of neurons from human TG sections. Considering that the diameter of a human neuronal nucleus is greater than the thickness of a 5-μm tissue section, it is likely that portions of human nuclei were excluded from the sections. This is evident by observing a neuronal nucleus on both of two adjacent sections under microscopy. By comparing the diameters of human neuronal nuclei with that of mouse neuronal nuclei microscopically, we found that the diameter of the mouse neuronal nucleus is about three-fourths that of its human equivalent. This could explain the difference in efficiency of neuronal DNA recovery, being 64% for human sections and 93% for mouse sections, because the 5-μm section contained a greater percentage of the mouse nucleus. Nonetheless, by two independent techniques, we determined that LCM/PCR detects the majority of the DNA content of a single neuron.
The sensitivity and specificity of the PCR assay for HSV-1 gG and VZV ORF62 sequences were first determined by analyses of serial dilutions of plasmid standards. We routinely detected ≥5 copies of target sequences in a 25-μl reaction mixture with or without carrier DNA when all HSV-1-negative DNA samples were read as negative. Assay specificity was then further determined by testing DNA from single neurons recovered from HSV-1-seronegative subject 701 in parallel with every assay of samples from HSV-1-positive subjects. When the cutoff threshold was set at <5 copies/reaction mixture, 1 out of a total of 119 independent reactions with DNA from subject 701 was read as positive (equivalent to 12 copies of gG1), being the only occasion, or false positive, detected from HSV-1-seronegative samples. These data suggest that, as performed, the LCM/PCR assay yields a real but very low rate of false-positive reactions.
HSV-1 genome copy numbers vary widely in individual PCR-positive neurons.From 119 to 235 single neurons were microdissected from sections of each of six study subjects and analyzed for HSV-1 DNA by means of real-time PCR with primers and a probe specific for gG sequences. Among 970 single neurons from five HSV-1-positive subjects (three of whom were seropositive, while HSV-1 DNA was detected from the total TG DNA of subjects 702 and 711, from whom postmortem sera were not available), 61 cells were PCR positive for gG sequences, with copy numbers/cell ranging from 5 (the limit of assay sensitivity) to 3,955 and a median of 11.3 genome copies/positive cell (Table 3). The majority of positive neurons harbored less than 20 copies of HSV-1 genomes: 43% had ≤10 copies and 28% had ≥10 but ≤20 copies per neuron. In 14% of the DNA-positive cells, however, the viral DNA copy numbers were ≥100, and a few of them were ≥1,000 copies/ cell (Fig. 3).
More neurons contain HSV-1 DNA than are found to contain LAT.As indicated in Table 3, the percentages of neurons positive for HSV-1 DNA ranged from 2.0 to 10.5% in the five HSV-1-positive subjects. This range of percentages is higher than the 0.2 to 4% of LAT-positive neurons we reported previously in human TG using ISH (11), and similar percentages have been found by others in their ISH studies of infected animals (4, 22, 44).
To determine whether the percentages of LAT-positive neurons in the TG studied now differ from those prior observations, sections adjacent to ones from which single neurons were captured for LCM/PCR were subjected to ISH using a biotinylated cRNA probe antisense to HSV-1 LAT and visualized with streptavidin-conjugated peroxidase and its substrate, TMB (Fig. 4). Even though some of our preparatory work (data not shown) suggested that the nonradioactive probe used here is less sensitive than the radioactive probe we had used in our previous ISH studies, we found that the percentages of LAT-positive neurons were similar to those previously reported (11), being 0.4, 1.2, 1.5, and 0.2% in TG of subjects 702, 703, 704, and 711, respectively. In these four subjects the percentages of their neurons positive for HSV-1 DNA were 2.0, 7.3, 10.5, and 9.8%, respectively. The percentages of HSV-1 DNA-positive neurons were 5- to 41-fold higher than those positive for LAT by ISH (Fig. 5). This finding strongly suggests that there are neurons with HSV-1 DNA detectable by PCR that are not ISH positive for LAT.
HSV-1 DNA is detected in LAT-negative neurons.To determine that HSV-1 DNA is detectable in LAT-negative neurons in human TG, as reported in mice (6), single neurons were microdissected from sections of human TG from subjects 702, 703, 709, and 711 after undergoing ISH for LAT. As shown in Fig. 6, HSV-1 DNA was detected from 4, 6, 11, and 7% of LAT-negative neurons from these subjects, respectively, while among LAT-positive neurons the percentages of cells positive for HSV-1 DNA were 27, 21, 16, and 40%, respectively.
A comparison of the HSV-1 DNA content in single LAT-positive neurons with that in LAT-negative ones is shown in Table 4. Since the percentage of LAT-positive neurons in these sections proved very low (e.g., 4/1,000, or 0.4%), most of the HSV-1 DNA-positive cells (e.g., 63/1,000, or 6.3%) did not accumulate LAT to levels detectable by ISH. On the other hand, the HSV-1 genome copy numbers in the positive neurons of these two groups were comparable, with medians of 18 and 12 copies in LAT-positive and LAT-negative neurons, respectively.
Detection of VZV DNA in human TG neurons.From 10 subjects, 146 to 209 single neurons were captured from their TG sections by LCM and analyzed for VZV DNA content using PCR primers and a probe specific for the diploid VZV gene 62 (Table 2). The percentages of VZV DNA-positive neurons ranged from 1.0 to 6.9%, with the median VZV genome copy numbers among positive neurons ranging from 4.5 to 29.8 (Table 5). Overall, a total of 4.1% (71 out of 1722) of the neurons were found to harbor 2.6 to 5,773 copies of the VZV genome, with a median of 6.9 copies per positive cell. Among the VZV DNA-positive neurons, 70% harbored ≤20 viral genomes while 9% harbored ≥100 copies (Fig. 3).
HSV-1 and VZV DNAs are rarely detected in satellite cells.To determine if HSV-1 and VZV DNAs can be found in nonneuronal cells, hundreds of satellite cells were microdissected from regions of the TG sections from which all neurons had been removed (Fig. 2). A portion of the DNA extracted from each cap was used to determine the cellular genome copy number, while the remaining DNA was assayed by PCR for gG1 or ORF62 sequences. In this manner, among 13 LCM caps prepared from sections of subjects 702, 704, 709, and 711, a total of ∼5,200 satellite cells were assayed and 21 copies of HSV-1 gG sequences were detected. In an additional set of 21 caps from subjects 701, 709, 710, 711, 714, and 726, in which a total of ∼14,200 cellular genomes were detected, only one sample was positive for nine copies of the VZV genome.
We developed and characterized combined LCM/PCR assays by which we could efficiently and specifically detect and quantify HSV-1 and VZV genomic sequences in single human neuronal cells. Using these assays on thousands of individual neurons and aggregates of nonneuronal cells, we have been able to further refine our understanding of the cellular distribution and estimates of the numbers of copies of latent viral DNA sequences in human TG cells.
Because LCM/PCR detects viral DNA while ISH detects LAT RNA, which is a highly stable and amplified species, the limits of their detection in our assay systems are dependent on the viral DNA load and the LAT accumulation levels in a particular neuron. Neurons that harbor copy numbers of viral DNA that are close to, or lower than, the sensitivity of the LCM/PCR assay may accumulate different amounts of LAT, ranging from a few copies to hundreds of copies of LAT per viral genome. Those neurons may not be able to be identified by the LCM/PCR methods for viral DNA, but those accumulating high levels of LAT can be detected by ISH for LAT. Thus, ISH may be more sensitive for detecting latency in such neurons. When the viral DNA load exceeds the sensitivity limitation of LCM/PCR, this method is more sensitive than ISH for LAT, as we verified by showing that the majority of HSV-1 DNA-positive neurons are ISH negative for LAT.
By using the LCM/PCR method, we determined that a mean of 6.3% of human TG neurons are HSV-1 DNA positive (Table 3). These numbers are lower than the percentages reported for experimentally infected mice, i.e., 7.3 to 32% by contextual analysis (45, 47) or 35% by LCM/PCR (6). It is most probable that the differences between what we observed in human cells and what has been observed in mice is due to the high virus inocula and route of infection used in mice, as have been discussed in the literature (45), and the short interval between infection and death in experimental mice, rather than representing technical differences among the assays employed. This explanation is supported by the independent observation in dissociated ganglionic cells that only some 3% of human TG neurons are PCR positive for HSV-1 DNA (5).
The HSV-1 genome copy number in individual neurons varied over 3 orders of magnitude (Fig. 3). The majority of HSV-1 DNA-positive neurons harbored, however, ≤20 copies of HSV-1 genome. Considering that the method we used was not able to identify neurons with less than 5 copies of viral DNA, the percentages of latently infected neurons and those harboring ≤20 copies of viral DNA should be even higher than we estimated here. On the other hand, 3% of the viral genome-positive neurons contained ≥1,000 copies of viral genome. This result is very similar to what has been found in mice by PCR of dissociated ganglion neurons (45) or by LCM/PCR (6). Our finding here by means of LCM/PCR that indicated that there are only a median of 11.3 HSV-1 genome copies/DNA-positive neuron proved to be concordant with what we ourselves estimated several years ago by real-time PCR of total DNA isolated from latently infected human TG, namely, that there are about 28 or more HSV-1 genome copies in LAT-positive neurons (39). Our findings here also are in concordance with estimates made by PCR of dissociated human TG in which HSV-1 DNA was found in 3% of the neurons and the viral DNA copy numbers ranged from 2 to 50 (5). It is not clear, however, whether the presence of higher levels of latent genomes in a cell indicates that they are undergoing, or are more inclined to undergo, reactivation. In this regard, we had shown in experimentally infected guinea pigs that higher total latent viral loads in a ganglion are associated with higher rates of genital disease reactivation (24, 29), but it was not known in those experiments whether more neurons were latently infected or whether neurons carried more copies of latent HSV DNA. It has been reported that in mice the HSV-1 strain KOS, which reactivates poorly, has less viral genome copies per positive neuron than the two other strains, 17syn+ and McKrae, which reactivate more frequently (46).
The combination of ISH and LCM/PCR techniques enabled us for the first time to analyze both LAT accumulation and HSV-1 DNA content in the very same neuron in human TG. By so doing, we demonstrated that the percentages of HSV-1 DNA-positive neurons in human TG were higher than the percentages of neurons that were found to be positive for LAT by ISH, consistent with recent parallel LCM and ISH studies of infected mice (6). In our human subjects, the percentages of DNA PCR-positive neurons were between 5.1 and 41 times more than the percentages of neurons that we found to be ISH positive for LAT (Fig. 5). In other words, neurons positive for LAT by ISH only represent a fraction of the entire pool of latently infected neurons. This finding may reflect a greater sensitivity of PCR for viral DNA than of ISH for LAT. The novel and more important message from our study is that in human TG not all latently infected neurons accumulate LAT to a high level, raising the questions of why and how the accumulation of LAT is regulated during latent infection. Given the observation in animals that HSV mutants impaired for LAT expression are able to establish and maintain latency in sensory neurons (2, 15, 37, 54, 57, 58), it is not entirely surprising to find latently infected human TG neurons in which LAT is not detectable by ISH. Our observations indicate that even in humans, the natural hosts for HSV-1, persistent substantial accumulation of LAT is not essential for HSV-1 to maintain latency. One is inclined to ask whether LAT accumulation is stable over the long term, which in the case of our human subjects amounted to decades from the likely times of their primary infections to death, and whether alterations, if any, in LAT accumulation are associated with periods of virus reactivation.
That our LCM/PCR method can only recover and reliably detect about 64% of the entire DNA content of a single human neuron and the fact that we detected HSV-1 DNA in only 16% to 40% of LAT-positive cells (Fig. 6) indicate that the percentages of latently infected neurons and the copy numbers of viral DNAs in neurons are likely even greater than we estimated from the LCM/PCR data. Moreover, it is reasonable to assume that there are latently infected neurons containing quantities of latent viral DNA below the assay sensitivity threshold of 5 copies/cell. Therefore, we conclude that HSV-1 latent infection in human TG is more pervasive than previously estimated by ISH for LAT.
HSV-1 LAT has not been detected in satellite cells in latently infected human or experimental animal TG by ISH, in situ PCR, and PCR of dissociated TG cells (11, 13, 34, 44, 45, 51-53). There was also no LAT ISH signal seen over nonneuronal cells in all of the sections in this study. Thus, the 21 copies of HSV-1 gG sequences detected in ∼5,200 satellite cells may reflect the very low level of false-positive reactions we identified in LCM/PCR studies and/or may be due to a low-level contamination with neurons amid the satellite cell samples.
Seventeen years ago, our laboratory reported that by ISH VZV RNA was only detected in satellite cells of human TG (12). Since then, the cumulative data from other laboratories have built a compelling case that VZV persists predominantly in neurons (17, 21, 25, 27, 28, 30). By real-time PCR of 1,722 single human TG neurons harvested by LCM, we detected VZV DNA in 4.1% of them. That we found VZV DNA in these few neurons is in accordance with recent data using dissociated human TG (28, 30) but is far less than the percentages reported by Lungu et al. using an immunohistochemical technique (32). This suggests either that the immunohistochemical method is far more sensitive than the LCM/PCR technique or, alternatively, that the immunohistochemical assays themselves may have yielded false-positive reactions.
Despite our prior report of VZV RNA in nonneuronal cells (12), we detected here only nine copies of the VZV genome in 14,200 satellite cells. It is not known whether this indicates that there is a small reservoir of latent VZV DNA in nonneuronal cells, spread of subclinically reactivating virus, or that the genome copies detected were due to the recovery of a satellite cell sample which could have been contaminated with very small numbers of latently infected neurons or represented the low level of false-positive reactions that we determined can occur in LCM/PCR studies. Nonetheless, we can safely conclude today that VZV persists in human TG predominantly, if not exclusively, in sensory neurons.
We thank David Kleiner and his colleagues for helping us harvest the tissue and serum samples, Thomas Fleisher and Rhoda Ashley for performing the serological analyses, Yo Hoshino for performing statistical analyses, and Jeffrey Cohen for ongoing discussions and critical reading of the manuscript.
This research was supported by the Intramural Research Program of the NIAID, NIH.
- Received 9 June 2005.
- Accepted 15 August 2005.
- ↵*Corresponding author. Mailing address: Medical Virology Section, Laboratory of Clinical Infectious Diseases, NIAID/NIH, Building 10, Room 11N-228, 10 Center Drive, Bethesda, MD 20892. Phone: (301) 496-7895. Fax: (301) 496-7383. E-mail: kwang{at}niaid.nih.gov.
↵† Present address: Miami-Dade County Medical Examiner Department, Miami, FL 33136.


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Item-Num.: CS0930
Summer Mens Tie in Beach Glass
Product Details
For a necktie that adds sublime elegance to your sharp suits and blazers, this Summer Men's Tie in Beach Glass is the perfect pick! A sunwashed shade of mint green is woven with silvery white threads for a boxed patterned texture, making it pop against shirts in black and charcoal. Team it with a dark gray collared shirt, a black blazer, and a silver pocket square for a charmingly sophisticated look. Handcrafted and jacquard woven from premium microfiber. Designed by Puccini.
Length: 58", Width: 3.25"
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Students will take a playing quiz on the Notes in Review starting on Monday, August 26th. Students will be graded on posture, correct notes, and their ability to keep a steady beat while playing. The quiz is 10% of their grade.
Welcome To Band! > 6th Grade Band > Playing Tests/Playing Quizzes >


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National Cancer Institute Guidelines for Exercise
In June 2010 a panel of thirteen researchers who are cancer experts came up with guidelines for exercise for cancer patients and survivors. The panel was formed to develop guidelines on exercise that were safe and effective. This is because there are currently over twelve million cancer survivors in the United States in need of advice!
Exercise is currently recommended for patients with breast, prostate, colon, gynecologic, and hematologic cancers, which pretty much covers all cancers.
The group was formed because the benefits of exercise were well documented for a number of cancers. In nearly all types of cancer, exercise was found to help with issues such as fatigue and physical functioning. These are factors that can directly influence the quality of your life. The guidelines wren also formed because it was discovered that patients on chemo or radiation could actually accomplish a lot more than anyone ever thought.
This panel also found that exercise was absolutely essential for a person's self esteem. This is especially true with individuals who have had surgery that alters their appearance. Activity can improve the person's body image. This is because changes in body composition are so typical with these types of cancers. Gastrointestinal, head and neck cancers can cause severe body wasting and exercise can help survivors of this type of wasting get their sense of self back and just feel more confident.
breast cancer can cause weight gain. In these cases the exercise can be useful for losing the pounds (sometimes caused by the drugs that are prescribed.) Breast cancer patients often need to build more body mass and lose some fat.
The idea that people undergoing chemo and radiation should be exercising rather than resting full time in bed is relatively new. At the NCI, which of course is a huge government operation, they are busy trying to figure out how physical activity can be integrated into cancer treatment and into the daily lives of survivors. Plans are also in the works for training professionals so that they can provide cancer-specific recovery. It is also being suggested that more exercise studios should be in hospitals and that mass workout sessions be part of oncology units. Investigations are also being made into the effectiveness of yoga and other gentler forms of exercise on cancer recovery.
There is no harm in making attempts to be fitter, whether that means taking a walk several times a week or going out to a gym to get some real training from a fitness professional. The idea is that once you do get our body in motion, you will start getting better; a fact that is being now well recognized by medical communities all over the world including the National Cancer Institute.
What is a Medical Fitness Trainer?
A medical fitness trainer is necessary if you want to get fit after an illness. People recovering from surgery or cancer definitely could use one. This type of trainer is also called a clinical exercise physiologist.
Currently exercise is recognized as a way of helping sick people recover from an illness faster. This type of guided physical fitness can also help you get through the symptomatic hell that often comes with being treated for cancer.
Working out at places like 24HourFitness can help decrease symptoms, eliminates the need for medication and also reverse conjunct conditions such as diabetes, obesity and heart disease. Aside from cancer, exercise is also beneficial people with depression, osteoporosis, cerebral palsy, chronic heart failure, fibromyalgia and other disorders. You can find coupons for 24HourFitness for a discount on a site like CouponKid.com
If you are a breast cancer survivor you might also be interested in the fact that exercise and a change of diet alone has been known to stop the development cancer right in its tracks.
To work out you do not necessarily need this type of trainer. Many ordinary fitness trainers are capable of putting you through the same type of training if you want someone to keep an eye on you while you are working out.
The reason that you want to work out is because "energy creates more energy." People who are ill and tired tend to feel even more exhausted after hours and hours of lying down. Ill people are often told to rest when really it is not in their best efforts to do so. If you lie in bed trying to recover it can lead to further weakening of your body and therefore a further health decline. The idea is to keep the balance and use the physical activity along with appropriate periods of rest to help your body recuperate. Stopping exercise to humor the disease is a big mistake.
Exercise as part of chronic disease management is gaining wider recognition. However when you don't feel well you may not feel as motivated to exercise. That is where a professional trainer can be of help.
A good fitness trainer will work with you and not push you too hard. He or she will be sympathetic to the type of chronic pain caused by cancer and its treatment and acknowledge if you are limited by fatiguing chemotherapy treatments.
It is widely recognized that a diseases like lung cancer or ovarian cancer is a formidable enemy but you have much more of a chance of stacking the odds in your favor if you are willing to go through a rigorous routine and then stick with the program.
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Please find below a list of selected hotels which can be booked through the hotel's websites or through standard booking systems.
Overview of the selected hotels
Please find bellow recommended accommodation during your stay in Vienna.
The conference venue is Wirtschaftsuniversität Wien marked in Red.
The hotels are placed in three categories:
- Hotels marked in Orange are 5-15 minutes walking distance to the conference venue.
- Hotels marked in Blue are 5-10 minutes walking distance to a U2 underground station which will directly take you to the conference venue.
- Hotels marked with Purple are in the central area of Vienna, where most cultural attractions are located. From these hotels you would need 15-30 minutes with public transport to get to the conference venue.


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Bernard Francis Boyle, Sr., died without warning, age 72, on Wednesday March 30, 2016 in Mesa, Arizona, where he resided with his loving partner Kay Goodfellow.
He was born August 28,1943 in Baltimore, Maryland.
He is survived by his daughter, Malissa Lynn Boyle, his 3 sons, Bernard Francis Boyle Jr., Michael Preston Boyle, and Christopher Scott Milbrett. He is also survived by his 8 grandchildren, and 4 great grandchildren, and we can't forget his best pal, Britches (his Boston Terrier).
Bernard was attending journalism college classes, made the dean's list, and was striving blissfully ""we are so proud "", he was also 30 pages into writing his first book.
Bernard will be laid to rest at the Veteran's Cemetery of Arizona, the family will be holding private services.
You are forever in our hearts…
Offer Condolence for the family of Boyle, Bernard FrancisPrint Obituary & Condolences


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SSPC /FROSIO Approved Training Licensee
Attend the job oriented SSPC / FROSIO training with HTS COATINGS. Upon successful completion of Coating / Painting Inspector training and certification program, student can work as follows:-
Coating or Painting Inspector
Anticorrosion Supervisor
Corrosion Control Engineer
QC / QA Inspector
Protective Coating Specialist
Painting Failure Analysis Expert
Present Challenges for Students & Professionals
Wrong selection of Academic courses
Inappropriate decision to select right core subjects
Inattention to Job oriented course for career growth
Inferior quality of Education in school and college levels
Not obtaining suitable jobs due to heavy competition
Upon placed, salary level is too low than expected
Selection of Job oriented course is always challenging
HTS Coatings will assist you to overcome all your challenges within a week
Get Certified with SSPC or FROSIO and work in Oil and Gas industries immediately
SSPC: The Society for Protective Coatings founded in 1950 as the Steel Structures Painting Council. SSPC is a non-profit professional society concerned with the use of coatings to protect industrial steel structures.
In 1997, the name of the association changed to SSPC: The Society for Protective Coatings. SSPC to better reflect the changing nature of coatings technology and the ever-expanding types of construction materials.
.SSPC is a 501(c)3 non-profit organisation focused on the protection and preservation of concrete, steel, and other industrial and marine structures. SSPC provides solution to the surface through the use of high-performance protective, marine, and industrial coatings.
SSPC is the leading source of information on surface preparation, coating selection, coating application, environmental regulations, and health and safety issues that affect the protective coatings industry.
SSPC PCI certified students could work as a Coating or Painting Inspector in anticorrosion industries anywhere in this world.
SSPC core products and services include:
Coatings industry standards
Training & certification courses
Painting contractor certification programs
Conferences and events
Technical publications
.FROSIO is a member organisation established in 1986. FROSIO has been introduced to overcome the growing demand of qualified and certified coating personnels in all the levels such as Management, Execution and Quality.
FROSIO acts through the formulation of quality requirements for surface treatment and Coating inspectors. FROSIO approval and certification scheme has achieved high international recognition.
FROSIO certified inspector is a specific requirement within different standards and specifications. Various levels of FROSIO certification is being issued to trainees, mid-level and higher level experienced candidates.
In India, HTS COATINGS is one only Training Licensee whom delivering FROSIO surface treatment and coating programs. Certified students can work as Frosio Coating Inspector anywhere on this planet.
When it comes to knowledge, HTS COATINGS is a recognised leader in Industrial Training.
With over 30 years of experience serving the Middle East and Gulf region, HTS founder has acquired the safety certifications, qualifications, and highly trained personnel to be among the safest coating professionals and specialists in our area.
Our company has created a robust program that has become the model for blasting and painting training in our industry.
As of 2019, we have trained more than 3000 sandblasters and airless spray painters. Presently they all working in India, the Middle East and Gulf region.
HTS approved by NSDC / PCSC / SKILL INDIA sectors and all students could benefit from our skills training programs in Chennai, India.
Please get in touch with HTS Coatings and we will provide numerous coating training related solutions to corporates, contractors and specialised service providers.
We have many modules for Coating, painting related skilled training programs such as Grit Blasters, Airless Spray Painters, Coating supervisors, Painting Foreman etc.,


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Sunday, November 12, 2006
Obsessive Moderate
I'm an obsessive moderate. I require balance.
I need the weather somewhere between hot enough to provide heat stroke and cold enough to be wary of frostbite. Both come rather easily to me. I've been on the edge of both and I don't appreciate edges. I prefer slow inclines and easy descents. Up and down is fine, as long as they are in smooth, relaxed motions. My mood tends to stay fairly stable; I'm not easily rattled, easily excited, or easily angered (more so than before, but military life and parenting and rude drivers in congested traffic will tend to strain patience levels). I need my schedule -- the routine that provides a comfort zone. It may be a lenient schedule, as a strict disciplinarian type, I am definitely not, but it matters to my mental state. During this schedule, I need a slow wake-up time, a productive "play" time, a time slot for some kind of physical movement to get the blood circulating, time to get business-type work done, and then time to do "my" stuff (most often that means writing time). I like to read before bed, pulling me from the "real" world into a transitional phase before I give in to the dream world. Interruptions in this balance I've created for myself will surely set me on edge. Too many of these interruptions will push me over the edge.
And you thought this month would be all about Nanowrimo.
It is. By my counter to the left, that little blue box that shows my progress or lack of, it's quite apparent I have more "lack of" progress than actual progress so far. There have been too many interruptions. I have time at night to write, yes. What I don't have is the balance in my brain that allows me to push other thoughts away enough to write. To be on track, I need to hit 20,000 words by tonight, which means adding over 10,000 words. I somehow don't see that happening. On good days when I do little else, I can write between 5,000 and 6,000 a day. Those days are rare. I average 1,000 words an hour when it's going well.
However, it's still quite achievable to hit the 50K mark by the 30th. I need only to average just over 2,000 words a day. Yes, quite achievable.
Yesterday, I relished in the luxury of a mid-November warm-up. Running around in only a light cotton blouse (and my jeans, of course) with the car windows down and the sun streaming upon my skin, I knew it was likely the last day of the year for such a luxury and therefore, absorbed it into my soul as well as I could. I took the photo shown in this post as well as a few others, appreciating the beauty of "my" trees.
Today, after a full night of streaming rain and the light howl of an occasional gust of wind, I look out and watch the leaves being torn from the branches amongst the gray of a wintery sky. Nature is indeed balancing itself. While I sigh over the lack of color that I will miss until spring, I enjoy the starkness of the tree trunks, their lines strong and graceful, branches dancing with the wind and resting with its calm.
And I know this is a good writing day.
Yesterday I played in the weather like the squirrels chasing each other up and down the trunks and leaping from branch to branch. Today I am settled in and ready to turn my attention to the solitude of the keyboard clicking and the coffee relaxing and invigorating me all at once.
"And the self-same well from which your laughter rises was often-times filled with your tears.
Kahlil Gibran
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Bonsai Art Wood Díszgalagonya Pink Flowers Plant
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