text:
Click for "Microbes After Hours" videos
Here is a transcript of TWiM episode #49, "Grape-like clusters". Thanks to Frank Shinneman for transcription.
The transcript is also available as a pdf file - click here to download.
Hosts: Vincent Racaniello, Elio Schaechter, and Michael Schmidt
Aired 16 January 2013
Vincent Racaniello: This week in Microbiology is brought to you by the American Society for Microbiology, at microbeworld.org/twim. This Week in Microbiology, episode 49, recorded January 10th, 2013. Hello, everyone. I'm Vincent Racaniello and you're listening to TWIM, the podcast that explores unseen life on earth. This is our first episode of 2013. Happy New Year, everybody.
Joining me today from Small Things Considered, Elio Schaechter.
Elio Schaechter: Howdy.
Vincent: Happy New Year.
Elio: Happy New Year to you and everybody who's listening.
Vincent: That's right. Another one. We started another year of TWIM among other things, of course. Also joining us today from the Medical University of South Carolina, Michael Schmidt.
Michael Schmidt: Happy New Year to everyone.
Vincent: Everyone have a good Holiday? Elio, you were in the desert right?
Elio: Absolutely.
Vincent: I have this vision of you standing in the desert with nothing around. But there must be houses or something there, right?
Elio: There's a town called Borrego Springs. But if you go a ways, you're right. Actually it's not like the Sahara, big dunes desert. There are some dunes around there but really there's quite a bit of vegetation in between. So if you look from an airplane you would probably see half-sand and half-vegetation. There's really little bushes. The creosote bush is the main one in the western desert. It's abundant, as well, but there's really, it is a desert by any definition.
Vincent: So do you walk around there? Nice to walk around, yes.
Elio: Yes.
Vincent: Michael, you were in Chicago, right?
Michael: And I was fortunate that the snow that was snowing didn't stick and I didn't have to shovel.
Vincent: You didn't want to shovel, huh?
Michael: No. Shoveling—my 91-year-old father still threatens to shovel and my brothers and I scold him for wanting to go out and shovel.
Vincent: Michael, You're a young guy. You can go out and shovel.
Michael: No. I would have gone out and shoveled.
Vincent: What are you 40 years old?
Michael: No, I'm older than that.
Vincent: I just became old.
Michael: Yes. Yes. I wish you a Happy Birthday.
Elio: I saw a T-shirt recently that said, "This is the oldest I've ever been". I can't argue with that.
Vincent: That's great. I like that. Elio wished me "Happy Birthday" by email. I said, "Well, I usually complain to most people but I can't complain to you." You would laugh at me, right?
Elio: Absolutely. Youngsters!
Vincent: We all look young, right. You look pretty young, actually.
Elio: Thank you.
Vincent: We have a couple follow-ups to last time. Actually, I have an email which I think I'll bring up as a way of introducing a follow-up to Jo's paper on subtle gender bias. We received an email from Robin, who writes:
The matriarch of a hunter-gatherer band might prefer males to be assigned to specialized training in hunting skills so the sub-conscious bias probably goes back a couple of million years to the early days of homo before sapiens. Who knows what the origin of the sub-conscious bias is.
Michael: True.
Elio: Let's just make sure that people who listening who may not have heard the last TWiM know what you're talking about. I wasn't here but I heard it afterwards and I was sorry I missed it in person because it was a fabulous TWiM. I mean both papers were incredible. And Jo presented a paper of her own, published in PNAS, which she did a remarkable little study that showed that in fact that there is an unconscious bias in people, both males and females. She did this—do you want to say how she did it, Vincent?
Vincent: This was specifically in science faculty. They identified 120 or so science faculty, different fields of science, at major universities, and sent them a CV or resume of a person applying for a lab position.
Elio: Lab manager.
Vincent: Lab manager position. They all received the same resume. Half had a male name and half had a female name on it: John or Jennifer. They were asked to rate this individual—would they hire them, how much would they pay them and would they mentor them. The results clearly show that the male applicant was vastly favored in all three categories over the female by both male and female science faculty.
Elio: One thing is, although the results are certainly statistically significant and cause for alarm, I don't doubt that, still numerically the difference was about 20% on average. Wasn't it?
Michael: Yeah.
Elio: Which means it's important. It's terribly important. But it not insurmountable. It's not like a factor of two or something like that.
Vincent: I think it can be surmounted and they suggested some ways of doing that.
Elio: Right. So it's a problem, and it's an important problem, but it's not a gigantic difference that they found. I was really taken by the experimental design. Experimental scientists, current or past, would have a hard time designing such an experiment and this was brilliant. Because it really had every factor conceivable that would ensure validity. I thought this was, in itself, quite dramatic. The ability that she showed, she and her colleagues. Jo worked with a bunch of social scientists on this, yes.
Vincent: Yes.
Michael: The leaders in the field, the ones that look at unconscious gender bias.
Elio: So they really put together a tricky but very, very successful instrument. They did, as Jo mentioned, have to be tricky. They didn't tell the recipients what the purpose of this study was about. The recipients were just told straight—"Rate them because we're trying to study how to best place our students" or something like that. It was something which was not quite true and they have to live with that. Jo mentioned this that she was uncomfortable with it. On the other hand, I think that most people of goodwill, at least, would not resent it at all and would say, "Well, this is what you have to do".
Michael: And they found out at the end. It wasn't like they were hood-winked. At the end of the survey they found out what actually was going on. So it's not like they found out well after the fact. They found out literally when they completed the survey.
Elio: It's probably an ethical question there. I wonder if this is any different than giving people a placebo or not giving a placebo and not telling them until afterwards. There's some vague analogy here that I perceive.
Vincent: And in both cases the only way you can get an answer, you have to have a placebo, right? And in this case…
Michael: You know in the new work for the discovery and testing of new antibiotics we can no longer give placebos because it's not ethically appropriate. It's non-inferiority trials that are being conducted today. So Elio's got a point in terms of whether or not we had─ she needed to look at something like a non-inferiority trial. I'll share with you one of the things that I did after that particular episode. I was so intrigued with the paper, when I got home I wrote an email to the dean of the College of Medicine recommending the paper that she would then recommend it to the chairs of the college and recommend it as a departmental journal club for the faculty to consider in terms of looking at this whole issue of gender equity. I also forwarded it on to the folks at my institution who are concerned with this use of gender equity and really encourage folks to take a look at the paper and to think about it. That's, after all, what journal clubs are for is to get people to begin to think. I think that was one of my take-homes from that really intriguing paper is to go out and become, if you will, an evangelist to say, "Hey, this is out there and we need to think seriously about it and develop a strategy to help our female colleagues excel".
Elio: Yes. Good for you. Did you get a response from the dean?
Michael: I got a response from the dean. She liked the idea and she forwarded it on to the committee that worries about those issues. They are currently noodling on that idea trying to figure out how to roll it out and see what happens. They have seminars that they often offer on this particular topic, all college seminars. So it may be something that gets presented or at least gets discussed.
Elio: I think it's great and I think we all should probably try to do something like that because it seems to me that awareness on the part of everybody, males and females, awareness of this being a problem is the first step. I hardly knew this. I kind of vaguely would have thought that there is an unconscious bias but I thought, "If we don't have a conscious bias, it should be able to take care of it," but in reality it's more pervasive than that. People should know about it. So let's all do that. Let's all send it out to various deans and people. Okay.
Vincent: Well, I blogged about it and I got a lot of quite interesting comments. I felt the same way, I wanted to do something in addition and my blogging platform reaches a lot of people, and Twitter and Google+, so I think it's important. I think you're right, Elio, that awareness will probably will go a good way although not all the way.
Elio: Right. Now I actually─ I liked what you wrote in your blog so much that I asked your permission to re-print it in our blog so it's going to have another shot in February, to keep the issue alive.
Vincent: I think that Jo, unfortunately, should go around talking about this.
Elio: She probably is.
Vincent: She is but she could do it an awful lot. She could spend her whole life doing that. But that really helps too, I would love to have her come here and give a talk on this. I think you have to let people know this. They're not just going to read just the paper. You have to emphasize it. Okay. Was there something else we wanted to a…?
Michael: I wanted to ask you, Vincent, since you're our resident influenza expert. I wanted to gauge your opinion. The press has been making a lot out of this year's influenza attack rate. There have been some intriguing hypothesis that have been promulgated in the scientific and non-scientific literature that one of the reasons the attack rate may be so high is because the mid-west was so dry this past year and that's why it's not attacking certain states. That hypothesis has appeared to have died over the last few weeks because the attack rate now looks pretty much countrywide. Would you care to comment on what you—because Boston is really having a tough time of things—in our local community we're restricting visitation and hospitals to only immediate family and so any thoughts or comments?
Vincent: Yeah. Interesting. Actually a reporter from USA Today called me this morning and asked me the same question. He said, "You're good at debunking myths. What's going on here?" First of all, the strain circulating this year, the predominant strain, is H3N2 and that's different from the last few seasons where H1N1 strains were circulating. H3N2 tends to be a little bit more lethal than H1N1. So we may see more deaths this year. It's too early to tell what the mortality will be. So that's one issue. There's no doubt about thatH3N2 is predominating and who knows why. We can't really tell why.
The onset of the first peak of illness or infectious—CDC does a wonderful job of collecting samples from all over the country and then doing diagnostic tests looking for whether they have influenza in them and, if so, what type it is. They have a wonderful website cdc.gov/flu/weekly. There, you can see every week they put of a graph and you can compare it to previous years. This year the onset of the first peak of flu is earlier than usual. I wouldn't say there are that many more cases than previously. For example, I'm looking the graph for 2010/11 right now and the peak number of positive specimens is about 6,000. That was in week six of the—which would be January, of course, so the peak there that year was a bit later. I don't know if we have peaked so far this year. We may have. If you look at the CDC numbers for this week the numbers are going down compared to the last few weeks but that is subject to revision. We could have another peak in January or so. So, we had a quicker onset. I don't' think the numbers are any higher.
If you look at New York City ─ I'm looking at a graph from the New York City Department of Public Health here. Now, this isn't as good as the CDC because this is just influenza-like illness surveillance data. These are people who have certain words like: "cough and fever" or "sore throat and fever" or "flu" and they say, "Okay, that's influenza-like illness but it may not all be flu." There is quite a spike in New York which is a little steeper than in previous years. Although in 2010/11 again there was quite a substantial spike of illness at about the same time as now. So my feeling is this is a little bit more serious of a flu year. It's nothing off the charts. There have been some recent years that have probably had as many or more cases. Maybe the onset is not as early but I don't think there's anything unusual. I think there's still time for people to get their influenza vaccines which they should do.
Now, unfortunately, as an example, I read an article on Forbes today, by a professor of epidemiology at Yale University. It's about Tamiflu, this antiviral for flu. He introduces this article: "Influenza-like illness is sweeping the country with CDC reporting that most areas of the country experience high rates". Remember that's still like influenza-like illness. It's not diagnosed again. So some of it may not be flu. "I should know", he writes, "my family is in the midst of it despite having been vaccinated". Now that makes people think: "Huh, this guy got vaccinated and he still got flu". But he might not have flu because there are other viruses that can cause an influenza-like illness or make you think you have flu. So just because you have the vaccine and got some kind of respiratory illness is no reason for others not to get the vaccine. Remember, the vaccine is not 100% effective so you might get it and still get influenza anyway. It's not the best vaccine that we have.
Elio: By the way, this year's vaccine that's going around against H3N2?
Vincent: Yes, in fact that's a good point. The match between the vaccine and the circulating strains is very good this year. They made a good choice which they don't always do. It's a hard choice to make. So that's my take Michael.
Michael: I think it's good. The watchword that I always tell folks about Tamiflu is that you have to take it within two days of onset of flu symptoms because afterwards the horse has left the barn and there's no point in closing the door because you just literally have too much viral load. The symptoms that you're going to feel or how you feel from the flu is just too far gone. It's no longer going to be effective because the way Tamiflu works is it prevents the virus from being released from the replicating cells. You sort of lock the virus in so it can't get out.
Vincent: The only thing Tamiflu will do is shorten the duration of symptoms. It's important to remember that when they did the clinical trial for that, that is what they measured, that's where they got an effect and that's what it's licensed for. So don't think Tamiflu is going to make you miraculously healed. It will just shorten the number of days that you feel lousy.
Michael: It's not penicillin, which is the generic concept of how antimicrobials should work. Because when penicillin was first released to treat bacterial illness it was truly a wonder drug that really stopped everything cold in its tracks.
Vincent: Right. Yes. Tamiflu's not a wonder drug.
Elio: By the way, Vincent, you did discuss this in TWiV, did you?
Vincent: This flu story? Not yet, no.
Elio: So when you do, our listeners of TWiM should go over to TWIV to hear the thing in better detail.
Vincent: Yeah, I don't know when. Maybe in the next few weeks. If we do it I'll just announce it on here again. I was thinking of doing an episode on this whole issue at some point. Because it's relevant now, obviously, people are worried. It's on the front page of the New York Times today.
Michael: It was on the front page of our paper.
Elio: Ditto.
Vincent: Okay. Let's move on to our two papers today. The first one has to do with Staphylococcus. Those of you who are faithful listeners should remember that not too long ago, episode number 42, we talked about this three-star pathogen, the meeting that Michael attended in Lyon, right?
Michael: Correct.
Vincent: So I remind everyone to go there and bone up on your Staph biology.
This is a paper from Nature. It is called: CCR5 is a Receptor for Staphylococcus aureus leukotoxin ED. It's from right here in New York City: New York University School of Medicine. This again has to do with Staph aureus. You all should know from listening to that episode this is a significant pathogen that causes all sorts of diseases: skin infections, soft tissue infections, bacteremias, pneumonia, sepsis.
The important aspects for this paper are that these bacteria make a variety of protein products that are secreted and allow the bacterium to do its nasty business: it makes super-antigens, it makes antibody-binding proteins, which we talked about last week, those proteins that bind—or two weeks ago—FC receptors of antibodies, FC portions of antibodies. They also make pore-forming cytotoxins. These are the leukotoxins that we're going to talk about today.
These leukotoxins are soluble proteins that are secreted and they recognize the membrane of a cell. Typically for these leukotoxins they're leukocytes or white blood cells like lymphocytes or macrophages. They make a pore in the membrane, basically make a pore, poke a hole in the cell membrane, this is the eukaryotic cell. That, of course, is bad for the eukaryotic cell and eventually the cell will die. Staph aureus makes four different of these pore-forming toxins. What we don't really understand about them is what they target on the host cell, what do they recognize. This is important information to know so you can understand the mechanism by which they work and maybe to design ways to subvert their activity.
So what they did in this paper, was to express these four toxins. They did three of them and they simply added them to cells in culture as a start. They say, "Do these toxins kill the cells?" It's very easy to tell if a cell in a culture is killed or not. There are a variety of assays. They actually used flow cytometry here. You can add the toxin, you wait a bit, and then you say, "Are the cells living or dead?" You can express that as the percentage of dead cells.
What they found, interestingly, with one of these leukotoxins, LukED, now this is a toxin composed of two protein subunits, the E-subunit and the D-subunit. It would kill cells, a T-cell line actually from humans, which expresses a chemokine receptor called CCR5. They had another T-cell line which did not have this chemokine receptor, CCR5, which was not killed by the leukotoxin. Alright, so two cell lines, both T-Cell lines from humans, one is killed by the leukotoxin that expresses CCR5. The other does not. It doesn't express CCR5. So that suggests, of course, that CCR5 is the target.
So what they did was to prove that they knocked the expression of CCR5 in a different cell line with small interfering RNA and they showed that, in fact, that confers resistance to this leukotoxin. On the other hand, if they start with the cell line that does not have CCR5 and then they express it that confers killing on those cells by the leukotoxin ED. And they also show that both subunits are needed and that none of the other chemokine receptors are able to make a cell susceptible to this toxin. So it looks like, at least in cultured cells, CCR5 is recognized by LukED. Now those of you who dabble in virology may recognize CCR5. I bet you do Michael.
Michael: I was wondering whether you were going to bring up our friend HIV to introduce…
Elio: You're friend! You're friend!
Michael: HIV is this really remarkable virus that I think has done more for immunology than any other virus because it's really helped us tease apart the immune system. Remember, during the pandemic phase of HIV, there was a lot of talk about CCR5 and delta 32. They go back to plague. They go back to smallpox in looking at CCR5. So I'm going to be very much interested in listening to what Vincent has to say about that.
Vincent: So CCR5, besides being a chemokine receptor on certain immune cells—and chemokines are proteins that are elaborated during an immune response. Their function is to recruit cells to the infected areas, say, so they bind receptors and that's how they exert their activity. CCR5 is one of these receptors. So it has a function in the cell. It also happens to be one of the two receptors that HIV needs to get into cells. So the most famous receptor for HIV is CD4 which is a molecule on T cells. But it requires a second protein, and there are two possible second proteins and one of them is CCR5. So there are some people, as Michael alluded to, that have a mutation in the gene for CCR5, it's called "delta 32". They don't produce this protein and they are naturally resistant to infection with HIV. There are about, I think, 10% of the people in the world have this allele this delta CCR5.
Michael: I thought it was only Europe. I thought it was only people of European extraction but I may be wrong.
Vincent: It is. So that 10% is basically all pretty much in Europe. We don't know what's selected for but as you suggest that some people think that it may have to do with the plague. There was once a hypothesis that smallpox selected for it but that's been debunked because the mutation's been around for a long time and in places where smallpox was not. We don't actually know what's selected for but it's in the human population. That will come into this story in a moment. Anyway, CCR5, because it's a co-receptor for HIV there has been a drug developed. There's several drugs developed that target CCR5. One of them is called "Maraviroc". It binds to CCR5 and it prevents HIV infection. They show in this paper that this drug blocks killing of cells by LukED, by the purified leukotoxin. So I think this is amazing that the drug that blocks HIV also blocks LukED killing. It's pretty neat. So far we've been working with purified proteins. Then, of course, they ask, "Does Staph aureus bacteria kill CCR5 expressing cells?" They have to jack up the expression of LukED in staph aureus because normally it's not made at very high levels. So they do that with a genetic trick. They find, in fact, that if you add those Staph aureus to cells expressing CCR5, those cells are killed. That killing can be blocked again by Maraviroc, that drug that binds to CCR5. If there's no CCR5 present, the bacteria will not kill the cells.
Michael: So this leads me to bring up the question of—you know, the epidemiologists were the ones who tumbled into this CCR5 and HIV and the correlation to Black Death or smallpox. So the immediate question that I have is, during those times, was it a co-infection when you had plague or when you had smallpox where you actually had frank staphylococcal aureus pneumonia, and it was CCR 5 and the Staph aureus was the true selective pressure rather than the Yersinia pestis or the Variola. Here we have these authors who, unequivocally, mechanistically demonstrate via the knockdowns with the silencing RNA as well as the delta 32 mutation which tumbled out of the work in the '90s with HIV. It really is a mechanistic argument driving home that Staph aureus does indeed use CCR5. If you're expecting natural selection to work on individuals before they're able to deposit their genes to the next generation, you can well imagine how getting deleted for CCR5 would be a selective advantage if you actually got a co-infection that compromised your lung and the staph sort of snuck in. But if you were deleted with CCR5, you may be able to survive plague.
Vincent: Yeah, I think that's a reasonable hypothesis. Sure. In fact, they say later that this adds now another pathogen that could have selected for the delta 32 mutation.
Elio: It's more common. Being it's more common that makes some sense, doesn't it?
Vincent: It could, yeah.
Michael: In the most recent New England Journal the Supreme Court has now said that it's possible to talk about off-label use of drugs because they're now—off-label discussions are subject to free speech. It's funny but true. The Supreme Court has decided that off-label discussions of drugs is subject to the free speech clause of our constitution. Maraviroc, which is used for HIV, could we use Maraviroc and some of those horrific cases where no drugs are working against the staph pneumonia.
Vincent: Well, I suspect someone's going to do a clinical trial, right, Michael?
Michael: Hopefully.
Vincent: Yeah, because it makes perfect sense. As we'll see in a moment it does have some effect in a model for disease. Now they showed very nice experiments that this leukotoxin, in fact, the LukE part of the leukotoxin binds directly to CCR5, so LukE but not LukD. Again, there are two polypeptides that make up this toxin. LukE binds directly with CCR5 at the end terminus and so this begins to give you some idea of how the toxins are targeted to the membrane of leukocytes. They bind directly to CCR5 and then, of course, that's followed by making a pore in the membrane. We also would like to know what cells are killed by LukED. So they do some very nice studies to identify the particular subsets of T-cells that are depleted when you add this leukotoxin. Again, it's important for us to understand the pathogenic effects of infections.
The last experiment is really striking to me. They ask in a mouse model for Staph aureus infection, what is the effect of removing CCR5. They have mice that lack the gene for CCR5. This is, as you might have guessed, from the delta 32 mutation in people, this is not an essential gene. They infect wild type mice and CCR5 null mice with staph aureus. The CCR null mice, the bacteria can get in to the infection site and they elicit an immune response but there is less bacteria in the kidneys of these animals and there's less mortality associated with infection in mice lacking CCR5.
Elio: Actually, this struck me so much. I was wondering if the title of the paper couldn't be a parallel title that says: Under some conditions leukotoxin ED is sufficient to cause death. It still is a difficult thing. Here's a multifactorial pathogen par excellence. Staph has half-a-dozen to a dozen major classes of what I call "virulence factors." In this case, under these conditions using mice, et cetera, it looks like that's all it takes. In other words, if you were to clone this toxin into an otherwise nonpathogenic organism, it could cause disease and kill mice.
Vincent: Yeah, I think that's a good experiment to do. Yes.
Elio: It would be. Yes, the kind of thing that Victor Nizet lab's likes in doing. Things like put the Lactococcus and puts the strep genes in it and stuff like that. It may not be sufficient but it comes close to defining a situation where you can make—tease out from this multifactorial, complex situation, you can tease out something relatively straightforward. It's not as simple as I say it but it would be very interesting thing to do. So the title could be: "Leukocidin ED is a major or, if not, sole virulence factor in staphylococcal infection." I don't blame them for not doing that, by the way.
Michael: It also suggests probably why folks making vaccines against Staph aureus have had such a terrifically hard time. If you think about it, if Staph aureus is able to infect T cells. T cells is a very important arm of the immune system, you can begin to understand how Staph aureus can literally take out that arm of the immune system with its toxin. As Elio alluded to, it may not be sufficient by itself, which is why Nizet has to do that fancy experiment, but it may suggest how staph is able to escape the vaccines where they've pulled out antigens and actually used them to vaccinate animals. In the animal, it works, but the animals don't have CCR5 unless it happens to be a humanized mouse. So I wonder—I'll have to go back and look at some of those vaccine papers and see what they used. Because, I think, this really will help the vaccinologist understand why the staphylococcal vaccines may have failed.
Vincent: Right. Basically to summarize this, this effect in mice, it's quite a substantial effect, about 80% survival as opposed to 20% survival when you have CCR5 present. So this toxin, LukED targets, remember, T-cells; it targets macrophages and dendritic cells, I didn't mention that. So basically if you take away the targeting of these cells the mice do a lot better so that tells us we have to look to see the role of these cells in protection against infection. The other interesting point they make is that the gene encoding this leukotoxin, LukED gene, is present in over 70% of strains that cause infections in the US and Germany. That's good. On the other hand, it is not present in some strains that cause hospital-acquired infections in the UK, for example. So it's not going to be the answer to everything, right, Michael?
Michael: Correct.
Vincent: It's going to be a good subset though perhaps. So you might consider treating some of these infections—I guess you'd have to type the infections for the presence of LukED before you would treat with Maraviroc, right, Michael?
Michael: Right. It would be the prudent thing to do. With the advent of PCR being so quick you could ask the simple question, does it have the LukED gene? You could just do a quick PCR and answer the question based on the strain. That's routinely done for a large number of viruses in the hospital today because they no longer culture viruses. But in the case of bacteria, a prudent use of an antimicrobial is always the key to preventing the emergence of resistance.
Vincent: Right. So that's what I think is quite a nice piece of work.
Elio: I thought so too. I was well quite taken. My bias about the title reveals that I'm on the bug's side and not the host's side. But it's really quite a paper.
Vincent: They may have tried to broaden the title but the journal may have discouraged them, for the reviewers, it's difficult, reviewers. Okay, let's move on to a topic close to all of us, wine.
Michael: Well since it's after the first of the New Year, I'll tell you how I discovered this paper. When I was first reading the CCR5 paper on Staph I did a quick search and I stumbled into another paper on community-acquired Staphylococci in PLOS ONE and this paper happened to be in the same issue. So I started to read this paper—and the paper I'm going to discuss is: The Vineyard Yeast Microbiome, A Mixed Model Microbial Map.
Vincent: You have to tell us where it's from Michael.
Michael: I am. It's from, and this is the dream job for most microbiologists. It's from the Institute for Wine Biotechnology at Stellenbosch University in Stellenbosch, Western Cape, South Africa. The paper is by Setati, Jacobson, Andong and Bauer.
Elio: I'm going to dare you, Michael, to make a connection between this paper and the previous one. Let's see what you can do.
Michael: Oh, god. The challenge flag has been thrown down. The easy one is, this is about grapes and if you take the name of "staphylococci" apart it means "grape-like cluster". That's the easy one. But, in reality, this is about vineyards. Anyone who has ever talked to an enologist, someone who is enamored with wine, they will use this fancy French word that I'm going to probably butcher in my pronunciation, so I apologize to my French colleagues, called "terroir."
Terroir is this remarkable concept. It has to do with the geography of where the vines are. It takes into account everything: the microbes that are in the soil, the microbes that are on the grapes, the microbes that are in the wine. It really takes into account everything because a lot of vintners use a natural crush where they don't add any yeast starter culture to control the fermentation of the grapes that then result in wine. This is a substantial economic issue for people who grow grapes for a living because you get paid on how well your grapes perform. That is, the wine that results from the crushing of the juice and the flavor, the bouquet, the complex odors that come out of wine that then are all a component of the flavor profile that we so enjoy if you happen to enjoy wines. You can do this experiment at home. You can take the famous bottle of Two-Buck Chuck that you can buy at one of those grocery stores out in California. In South Carolina we don't have any two dollar bottles of wine. That's Elio's advantage living in California. He can get a decent $2.00 bottle of wine.
Elio: Actually, they are selling in quite a few other places, as well, in the east.
Vincent: Yeah, we have it in the east but it's three bucks now.
Michael: Yeah, unfortunately, South Carolina we don't have anything as inexpensive.
Vincent: But I could send you a case Michael.
Michael: I think that's illegal in South Carolina too—sending, mailing wine.
Vincent: My name is Racaniello.
Michael: The bootlegger! So I was really intrigued by—and again this is the proper crafting of a good abstract. As I was reading the abstract, the authors used this term "Theory of Sampling". We know that terroir is important for wine but the other thing folks should appreciate is terroir or...
Elio: Try terroir.
Michael: Terroir, there's no L in it. Terroir is important is consider coffee and consider chocolate. Something that most folks cannot live without. Chocolate is also similarly a natural fermentation that takes place in the rain forest and we don't even have a starter culture for making chocolate. It's a natural fermentation and it's one of the principal reasons for preserving the rain forest is if we lose the rain forest we lose chocolate because there is no starter culture for chocolate while there is a starter culture for wine. It has all sorts of other applications. In thinking about this, the authors, because the principal point of this paper is they're asking a very straight-forward question: Does the quality of grape that they will then use in wine, is it influenced by the farming practice? They evaluated three farming practices. The first, of course, is conventional farming where pesticides may be involved. The second is biodynamic, which is their phrase, which I think in the United States we can probably appreciate as being organic where they don't use any commercial or chemical-type company insecticides. They use compost and they use additional minerals to augment the soil. Then they used a third method of farming called the "Integrated Production of Wine," which is a mixture of organic farming as well as the conventional farming. There is no set of codified rules that say, "You must do this. This and that." Basically their Integrated Production of Wine results that they use mulches and oak cover crops to improve moisture and soil fertility. They have bait traps to accumulate insects and they used ducks of all things…
Elio: Used Ducks. Yeah I like the duck part.
Michael: Yeah, to go around and eat the system. Now imagine their problem. The first rule of agriculture, that I learned, is that farmers are barely making ends meet. So consequently these folks don't have a lot of money to do an exhaustive sampling of their territory. They're evaluating plots of land in the same geographic region and they're asking the question: Does the microbial diversity differ based on the farming. The way folks traditionally determine whether or not you have reached saturation is you sample until you see a plateau in the number of species that you're sequencing your 16S, your 18S, whatever you happen to be using, reaches that plateau. Which, as you can well imagine, would be very expensive because how many sequences, how many bacteria do you have to isolate, how many yeast do you have to isolate in order to recognize whether or not you have a representative sample. So these authors did something brilliant. They employed the theory of sampling. Without getting into too much math you can go into their paper and they explain the math in gory detail but I'm going to try a metaphor. Since we just finished an election cycle in the United States, did you ever wonder how the polling people know who are going to win an election? What they do is they survey likely voters. They employ this theory of sampling. What they do is they take a particular geographic region and they convert it into a two-dimensional plot and they run it through this algorithm that effectively tells them where to sample. In the case of Vincent's neighborhood, where people live at many different levels, they tell you which floor to sample. Now imagine them. They're looking at grapes. Grapes don't just grow in the ground like a potato. They grow at all different levels, they may or may not have leaves shading them and the yeast that…
Elio: The usually grow on trellises, don't they?
Michael: They grow on trellises. But some of the trellises may be in full sun. Some of the trellises may have leaf cover where they're covering the grapes. What these authors did is they used the grape as the perfect collection vehicle. They had to understand where to pick the grapes. So they had five trapezoidal plots of land that were evaluating these three different farming methods. What they did is they linearized them so that they could then determine in incremental sampling. What happens is the the computer spits out a 2D map telling them at what height and at what location to accumulate the grape. They then accumulated the grapes. They rinsed them in normal saline and there was a comment. The neat thing about PLOS ONE is people can write into the authors in the comment section and say: "By using saline aren't you osmotically stressing the yeast and killing them?" These authors simply looked at yeast. They didn't care about bacteria in this instance because what they were looking at are the epiphytic yeast. An epiphyte, of course, is just something that is growing on something. It's not actually eating the grape, it's just on the grape. The yeast are somehow making their living out of the air sort of like an orchid.
So they asked the first question of their paper is: Did the farming methodology change the density of the bacteria based on the farming practices?
Elio/Vincent: Yeast.
Michael: Yeah, excuse me. I got bacteria on the brain today. The yeast. They did a really nice job of showing us the concentration of the bacteria.
Vincent: Yeast.
Michael: Yes. Keep correcting me. The yeast and the concentration was not that high. It was between—the total yeast population ranged between 4 – 8 X 104 CFU per gram of grape. That was found not to be statistically significant between the different farming systems. The way they asked about the genomic differences or the differences between the yeasts is they did it in two methods. The first is the old-time religion. They cultured them then they identified them via traditional methodology for identifying yeast. I got it right that time. What they learned is that they didn't find all that much of a difference between the cultivars. They report in their paper that in the conventional farming they had one predominant species, Aureobasidium pullulans, was principally there at 70% distribution. The Integrated Farming method, which if you are thinking intuitively about complexity, you would expect that it would be a little bit more complex than the conventional farming. Conventional farming methodologies because the thought is that harsh chemicals and insecticides will limit biodiversity. You would expect that predominance of one species intuitively would go away and they did indeed see that. The Organic methodology, or the bio-diverse methodology, or Biodynamic method of farming you would expect to be more heterogeneous. They then further asked the question whether or not this was indeed the case. They used a methodology called "ARISA". ARISA is "Automated Ribosomal Intergenic Spacer Analysis". Now, we're all familiar with the bacterial ribosomal RNA operon in that there is a 16S gene and then there is a spacer gene and then the 23S gene. Since yeast are Eukaryotes, they did a variation of this for fungi and there is tucked in-between the 18S ribosomal RNA in yeast and the 28S ribosomal RNA in yeast, there's the 5.8S ribosomal RNA and they have a fluorescently-tagged 5-prime primer. This 5.8S ribosomal RNA region is amplified and it gives you a different sized piece. Since one end is fluorescently tagged, they run it either on an agarose gel or they were fortunate in they had a capillary gel electrophoresis system that has a specific detector in it that fluoresces and it literally gives them a read out of how big the region between the 18 and 20S gene is. They can determine the complexity of the population. Of course, they can go on to sequence it. What they learned is that there were, as you can well imagine, as they ran this through the statistical packages that are now quite common in any laboratory that does DNA sequencing, they asked the question about species richness and species evenness and then species diversity. I hadn't looked at the math on these subjects in a while and I was really intrigued about how clearly this paper helps me visualize whether or not their sampling technique really worked. You look at the Melnyk's Index of Species Richness and the conventional farming gave them a 0.96, the intermediate form of farming gave them a 1.06 and then the biodynamic gave them a 1.45.
Species richness is really an interesting concept because what they do is they simply ask the question: How close in numbers each species in an environment are? The math is really pretty complicated but if you think about—and the example I tumbled into when I was trying to figure out how to explain this so I would survive my good friend Elio's telling me "Make the math simple". So if there are 40 foxes and 1,000 dogs the community is not very even. But if there are 40 foxes and 42 dogs then the community is quite even. The evenness of the community is then represented by that Pielou's Index of Species Evenness. The working hypothesis is that conventional farming should have a less even coefficient at 0.5 while the organic, which is at the other extreme, should be less even. It is, indeed, at 0.76. The inverse of that is the diversity index or the species diversity. What they found is the conventional had a species diversity index of 1.2 telling you it's not very much diverse. The intermediate was 1.45 and then the bio-dynamic version was 2.15. The reason I brought this paper to TWiM is principally because I think papers like this will help people begin to ask the question about human microbiomes. How do you know if something is remarkable? Recall back to that TWiM where we asked the question about the bacterial link between autism and traditional gastrointestinal upset. And we found the Suterilla species in the autistic population, but it was a very small sample size, only 27. What this sampling theory will allow us to do is teach us how to answer the question whether or not we are indeed seeing a representative sample. This is important for anyone doing community analysis. I was very much intrigued by sampling analysis because I'd been working in the area of hospital-associated infections and we've been sampling the built environment of the hospital. Trying to understand how bacteria move in the hospital, how antibiotic resistance moves. So how are you going to be able to afford to do large scale studies when molecular biology is not cheap? Think about sequencing. Even though it's come down substantially in cost, you're going to have to sequence quite a few if you use the traditional modality where you reach a plateau in finding something new. Whereas if you could use the power of statistics—you know, the polling folks have used that to really forecast elections ± 2% or ± 5% in order to predict the outcome of an election. You know the census folks have similarly been arguing that rather than doing a traditional head count of everyone in the United States, they can use statistical methodology in order to arrive at how many toilets are in a particular geographic location.
Vincent: So Michael, basically, when you're looking at large complex populations you use these mathematical methods to allow you to compare, in this case, different farming methods, without having to enumerate every single member of the different populations. Right?
Michael: Yeah, because the way you would do it traditionally is you would literally go in there with a harvester and take every single grape and literally sequence the yeast epiphyte population from each grape.
Vincent: So this you do it by sampling specific ones.
Elio: It goes beyond it. Let me tell you why I'm so happy that we did this today. It illustrates what you've just been saying that statistical methods can, in fact, be used throughout. They can be used to answer questions. But the thing it really emphasizes is the analogy of the question. In other words, you can ask, in microbiology, questions clear across fields and they come out to be the same. These are population structures in certain environments and how they are influenced by modification. So this brings up what I think is the power of modern microbiology. We teach a course—in an hour I'm going to be going over there—Jo Pogliano and I at UCSD, teach a course, we call "Integrative Microbiology". We think microbiology is an integrative science. It integrates prop questions that pop up from all kinds of different places. So I think this is delightful that we have been able to deal in—the same people, I mean, who are able to talk about something which sounds as disparate as Staphylococcus aureus and yeasts on grapes. I think this is the power of modern microbiology really.
Vincent: By the way Elio, we did you friend Jo's paper on TWiV not too long ago. You should tell him that.
Elio: Did you?
Vincent: Yeah, one of his papers about phage making microtubule in host cells. Just tell him that we did it.
Elio: Okay. I'll tell him.
Vincent: So Michael, this is the cool take-home message but in the end they wanted an answer to their question about the variation between the methods and what is that?
Michael: The take-home message is what you would expect. Is that the organic farming did indeed give them a little bit more complexity and the culturable yeast population, you know the god-fearing old-time religion culture, did not reproducibly predict the fully representative cohort that was present. It's only through this molecular ARISA analysis that we're able to discriminate amongst farming practices. The yeast distribution was significant and it was subject to significant intra-vineyard spatial fluctuations. The frequent heterogeneity of tank samples of grapes harvested from single vineyards at the same stage of ripeness might therefore be at least due to these different microbiota in different sections of the vineyard. So looking at this from an economic perspective in how the farming industry is going to take advantage of this paper, they're going to literally do ARISA on grape lots where they will go out to a field and they will develop an index to understand how much to pay the farmer for his grape crop based on what type of wine that particular ARISA pattern turns out. The analogy that I thought when I was reading this, is this is very similar to what the dairy industry used to do for farmers. They would take a sample of raw milk from a dairy farmer and they would add methylene blue to it. It would immediately turn milk, this blue color, the stuff that you saw on the first Star Wars movie when Luke Skywalker was drinking blue fluids. Then they would start a stopwatch. Based on the bacterial load in milk, the milk would go from that Star Wars blue to milky-white. The faster it turned white, the less the farmer got paid per pound for his milk because it had…
Elio: No kidding? That's funny. That's great.
Michael: It had greater bacterial contamination. Moving this into the modern era, I can imagine this happening in California immediately because this technology, all you need is capillary electrophoresis system, two PCR primers, PCR up, you know, grab the grapes based on the farmer's plot. You can then assess whether a Cabernet Sauvignon is going to make you a $100-bottle of wine versus a $2-bottle of wine. So there is real economics here and it's up to the, again, enterprising microbiologist to go out and do this mapping but it's all based on this sampling technique.
Elio: Michael, you're making a jump here. You're making a jump that you're going to tell the quality of the wine from the composition of the yeast population. Is that, in fact, true? Can you make that jump?
Michael: Well a lot of commercial wineries are now using starter cultures to overwhelm the microbiome that comes with the grape.
Vincent: Yeah, then you can control it. Right?
Michael: Then you can control it. Our friends out at University of California at Davis have a whole department of microbiology and enology where they actually marry the two of these things together. So I'm sure we'll get letters from them if I got it wrong.
Vincent: Well then they can come on and set us straight.
Michael: That's right.
Vincent: So, in France, where the best wines are made, do they used to…
Elio: Well?
Vincent: I know California's pretty good.
Michael: South Africa too.
Vincent: But do they use starter cultures—is that considered okay to starter culture? Is that blasphemous?
Michael: It's not blasphemous. The brewing industry, of course, no longer lets things to chance. After all most of the baking yeast that we buy in the grocery store to make bread comes from breweries.
Vincent: I guess you could say if you get a great bottle of wine it doesn't matter how you do it. Right?
Michael: That's right.
Vincent: That's really cool. I think we ought to get a person from this industry to tell us all about it one day. That would be fun. Then they could send us a case.
Michael: That would be wonderful.
Vincent: You know, are you done with that so we can move on?
Michael: Yes. I'm done.
Vincent: We have actually an email which is totally relevant to this so let me read it. This is from Chip. Chip writes:
"Saw this paper in my inbox this morning and thought It might be a good fun discussion paper on an upcoming TWiM."
So this paper is called: "Brewhouse-Resident Microbiota Are Responsible for Multi-Stage Fermentation of American Coolship Ale." So this is beer. American Coolship Ale is a type of beer that we make in the US. It uses production methods developed in Belgium by the Lambic brewers. Traditional Lambic fermentation is done using all the microbes in the worts. They expose the worts to the atmosphere overnight in a vessel which is called a "Coolship." That's why it's called Coolship Ale. It becomes inoculated with autochthonous yeasts and bacteria that then, over the next one to three years, carry out the fermentation in oak casks. This is a kind of beer, Michael, where you use the endogenous yeasts and bacteria, you don't use a starter culture. I guess it's a traditional kind of Ale. So they wanted to know how this culture develops over time. They used a bunch of methods. They used restriction fragment length polymorphism, barcoding sequencing, quantitative PCR and culture dependent analysis. They traced over the time of the fermentation, it's a couple of years, the sequence of populations that arise. For example, in the first month, they get Enterobacteriacea, a bunch of oxidative yeasts, then they get Saccharomyces spp. and Lactobacillales. Then a year later they get Brettanomyces bruxellensis, and then Candida, Pichia. It's just amazing.
Michael: Pichia, which is a biotechnology wonder, wonder yeast.
Vincent: Right. Use it for expression. So they define this whole sequence of events that happened in this culture. I guess the idea would be that maybe you could use this information to make sure things are going properly.
Michael: I think you could go and figure out how to sample the floor of the brewery where this is made so that when you make another brewery you can answer the question of whether or not—you know the whole issue of brewing is to turn out a consistent product.
Vincent: Right. So you're depending on the microbes coming from wherever you have this Coolship sitting, right? Atmosphere, so figure out ─ maybe from this they can from this figure out a starter culture to take care of it more consistently. I never heard of Coolship Ale. Have you?
Michael: No.
Vincent: Well it comes from the Belgian Lambic brewers. That's pretty cool. Alright, let's do one more email. This is from Meimei:
"I do not have the talent for a M.D-Ph.D. at Columbia, but I am definitely a huge fan. It's striking how and where the interaction of antibody and bacterium occurs (TWIM #48)! Learned a lot from listening to your interpretation of the paper. Many Thanks! Just one question about one antibody and one bacterium. Is it commonly considered more than one antibody molecule per bacterium?"
The answer is yes. If you looked at the images in that paper there are multiple antibodies bound to each fiber, actually, on each bacteria in one or the other orientation.
"P.S. Can this finding extend to the interaction between virus (e.g. HIV) and neutralizing antibody? It might be we just look at the wrong place for all these years!"
So he's asking whether antibodies can bind to viruses via their FC receptors, right, just the way they bind to bacteria by FC-receptor-binding proteins. So it turns out that there is a virus, Herpes simplex virus, where the virions on their surfaces have FC receptors. Those are believed to bind antibodies as a way of evading immune responses, just like the bacterial FC-binding proteins are an immune evasion mechanism. So it's believed that that's what Herpes simplex does too. It's the only example I know of so it seems to be less common amongst viruses, but it does happen.
Michael: And the problem, in the case of viruses, is they do all their replication inside the house, inside our cells. So you would have to have an antibody level at a high enough level at all times to effectively—because in HIV the whole conundrum is you need to prevent infection rather than disease, which most vaccines try to do this. They prevent the disease from happening rather than the infection. That's why it's been so challenging to develop a vaccine for HIV is that it must prevent the infection because of the cell that HIV infects. The immune system and it takes it out. So it's a really good question.
Elio Schaechter is at Small Things Considered. Thanks for joining us Elio.
Elio: Sure. I missed the last one. I must say this was pretty unusual and I enjoyed it a lot. Thank you very much everybody.
Vincent: Michael Schmidt is at the Medical University of South Carolina.
Michael: Thank you Vincent. Elio, I'm glad I was able to survive the math.
Vincent: You did a great job.
Elio: You did nobly. You really did.
Vincent: You know I think the title for this could just be Grape-like Clusters.
Michael: Grape-like Clusters. That is true.
Elio: Hey, not bad.
Vincent: It will be fine. I'm Vincent Racaniello. You can find me at virology.ws. Many thanks to the American Society for Microbiology for their support of TWIM. Chris Condayan and Ray Ortega for all their help behind the scenes. In case you're wondering, the music you hear at the beginning and the end of TWiM is composed and performed by Ronald Jenkees and we use it with his permission. You can check out his work at ronaldjenkees.com.
Thanks for listening everyone. We'll see you next time on This Week in Microbiology.
- End –
Transcribed by: Frank Shinneman


text:
Recording Residential School
Ian Stewart/Yukon News
The Truth and Reconciliation Commission wants to establish a museum for Canada's residential schools.
In 2008, Prime Minister Stephen Harper apologized to Canadian aboriginal communities for more than two decades of forced assimilation of First Nation's culture through the 130 schools.
The fallout of that federal-sanctioned policy has affected generations of people.
A year after the apology, Harper signed the settlement agreement which established the five-year mandated Truth and Reconciliation Commission.
Its job is to educate Canadians about what happened to the estimated 200, 000 aboriginal Canadians forced to attend the schools.
The commission has been touring communities and holding events to gather and record peoples' own stories, memories, artifacts and art.
Last week, the commission announced it wants to house all this material in a research centre.
In March, experts and survivors will gather in Vancouver to decide what the centre should look like and how it should work.
"It will be a permanent resource to educate all Canadians on what happened," said commissioner Chief Wilton Littlechild on Wednesday, before inviting researchers, students, leaders and survivors to attend.
The Vancouver forum is a chance to hear from experts on the best way to do this, he said.
And while little has been decided about how to go about creating this centre, a glance at the agenda and speaker list gives clues about the intended direction.
The speakers are coming from a wide variety of countries – South Africa, Rwanda, east Timor, Serbia and numerous countries in Latin America – that share one thing, a genocide or ethnocide in their history.
The proposed Canadian residential school centre will show the personal history and experiences of residential school students and how the system was, in fact, conducting an ethnocide.
"I can't see much basis for it helping in anyway," said Chuck Hume, a residential school survivor from the Champagne/Aishihik First Nations.
He now works as a resolution health support worker with the Council of Yukon First Nations, helping other survivors accept their past so they can deal with their future.
"You got to remember a lot of that material that's given out is quite confidential and I think it should be kept in kind of a private way," he said.
Personal accounts of kidnap, psychological, physical and sexual abuse tend to take over most of the commission's visits and gatherings.
If survivors' children wish to read about this in their family members' own words, for example, that should be made possible, said Hume.
The forum is set to take place at the Sheraton Wall Centre from March 1 through to the 3.
Information and registration is available on the commission's website at http://www.trc.ca.

subset: commoncrawl


text:
Informazioni importanti
- Short course
- In 50 sedi
- Durata:
5 Days
- Quando:
ZZahid Momin
Il meglio I would like to say some words, of course it was a very pleased moment for me to pass this certificate, to qualify for this certificate and of course, this is not just a certificate for me. During these days, I have erudite a lot from Dr Crawford and of course, thought I have a very long eleven years of experience in HR and OD but I would concede and I would reconcile that many of the concepts and discussions made by DR Crawford with me were new for me and some of them I realized that those aspects are Art of OD and I have a firm amateurism that I will retrieve these motifs and new systems taught by him.
Da migliorare Everything good.
Corso realizzato: Febbraio 2016 | Consiglieresti questo centro?
Da migliorare Everything good.
Corso realizzato: Febbraio 2016 | Consiglieresti questo centro?
Cosa impari in questo corso?
Communication Skills
Organizational Skills
Team Development
Time management
Communication Training
Effective Communication
IT Development
Skills and Training
Team management
Organizational Improvement
Organization development
Organizational Analysis
Organization Management
Course Contents, Concepts and Issues:
Organisational Development
Organisational Development
- What Is Organisational Development (OD)?
- OD and Organisational Effectiveness:
- Differing Perspectives of Organisational Development
- Organisational Climate
- Organisational Culture
- Organisational Norms
- Organisational Values
- Organisational Power Structure
- Worker Commitment
- Structure of Roles in Organisation
- Inter-Group Collaboration
- The Combination of the Authority Based in Roles With the Authority Based in Knowledge and Skills
- The Creation of an Open System of Communication –Vertically, Horizontally, Diagonally; Management Development
- Micro Organisational Development
- The Quality of Working Life (QWL)
- Aspects of Quality of Working Life:
- Adequate and Fair Compensation.
- Healthy and Safe Working Conditions.
- Development and Growth of Human Capacities
- Growth and Security
- Social Integration of People
- Constitutionalism
- Protection of Total Life Space
- Social Relevance of Work
- Sensitivity Training
- Approach To Organisational Development
- Organisational Development Interventions
- Process Consultation
- Enhancing the Effectiveness of Programmes
- Macro Organisational Development
- Determination of Success
- Influence Change Strategies: When They Should Be Used or Avoided
- Control Change Strategies: When They Should Be Used or Avoided
- Communicating Organisational Change
- Communication Media: Mass or Personalised Communication?
- Mode and Channels of Communication
- Getting the Message Right
- Timing of Communication.
- Who Should Communicate What, When?
- Use of Groups in Change Process.
- Managing Latent and Manifest Resistance to Change.
- Effective, Overall Change Leadership
- Leading Change Implementation.
- Selecting the Appropriate Change Agent
- Internal or External
- Speed of Change
- Change Acceleration: Averting Organisational (Organisational) and Individual Casualties
- Confidence
- Change Tolerance and Individual Stress Levels
- Managing The External Environment: Improving Perception and Instilling
- Stakeholders, Generally.
- Shareholders and Funding Agents
- Customers and Clients
- Potential Customers and Clients
- Change Institutionalisation: Returning To Normality
C'è qualcosa che non ti convince in questo corso? Segnalaci se ci sono errori e aiuterai altri utenti come te.


text:
Archived Honors & Awards 2013
- P. J. Maziasz, AIME Champion H. Mathewson Medal Award, for award winning paper, "Structure and Composition of Nanometer-sized Nitrides in a Creep-Reistant Cast Austenitic Alloy, Metall. Mater. Trans A. vol 41A, Dec. 2010, 2014.
- Amit Pandey, has been selected to receive the 2014 TMS Young Leader Professional Development Award (Nov. 27, 2013).
- Randy Nanstad was awarded the Distinguished Engineer, on ORNL Awards Night for sustained and major contributions to the broad area of nuclear materials technology, impact in the field of fracture mechanics of light water reactor pressure vessels, and a key role in preserving HFIR as a national scientific resource (Nov. 2013).
- Ryan DeHoff was awarded the Early Career Award for Engineering Accomplishment, on ORNL Awards Night, for exceptional accomplishments and leadership in developing the additive manufacturing research thrust area at ORNL (Nov. 2013).
- Bruce Pint has been selected as a NACE 2014 International Fellow (Nov. 11, 2013).
- William Peter was awarded the "40 under 40" award, by the Greater Knoxville Business Journal recognized as East Tennessee's ext generation of leaders in business and the community. Honorees will be featured in the magazine's January 2014 issue (Oct.14, 2013).
- Amit Shyam was selected as one of the winners of the 2013 TMS Micrograph Project, Apparatus, System, and Method for Detecting Cracking Within an After Treatment Device, "Silver Cermet" (Sept. 10, 2013).
- Steven J. Zinkle, Nuclear Materials, an authority on the effect of radiation on materials in fission and fusion nuclear reactors, has been named the 13th University of Tennessee-ORNL Governor's Chair. He was appointed( Oct.1, 2013).
- Edgar Lara-Curzio, was elected to the ACerS Board of Directors, (Aug.19, 2013).
- Nancy Dudney – was elected as Electrochemical Society (ECS) Fellow (Aug, 2013).
- Mikhail Sokolov receives ASTM International Award of Merit for Contributions to Nuclear Technology and Fatigue and Fracture Committees (July 10, 2013).
- Mikhail Sokolov selected as ASTM International Fellow (July 10, 2013).
- ORNL Wins six R&D 100 Awards - ORNL Today, July 7, 2013 - SYMMETRIX HPX-F Nanocomposite Separator for Improved Lithium Ion Battery, was jointly developed by Porous Power Technologies LLC and ORNL. ORNL's participants are David Wood III, Claus Daniel, Wallace Porter, Amit Shyam, Cristian Contescu, Rosa Trejo, Edgar Lara-Curzio, Jane Howe, Harry Meyer III, Jianlin Li, Ralph Dinwiddie, Curt Maxey, Hsin Wang and Beth Armstrong. SYMMETRIX HPX-F addresses market demands by lowering lithium ion battery costs and improving safety through the replacement of polymer separators.
- Federal Laboratory Consortium (FLC) National Excellence in Technology Transfer Award winners included, Pulse Thermal Processing, developed by the team of the following scientists and staff: Chad Duty, Craig Blue, Art Clemons, Nancy Dudney, David Harper, Ron Ott, John Rivard, Adrian Sabau, Alex DeTrana, and Stan Farnsworth, Kurt Schroder, and Charlie Munson of NovaCentrix.
- Yutai Katoh elected as an American Ceramic Society (ACS) Fellow.
- Shawn Reeves - awarded the 2013 Outstanding Technologist Award in Physical Sciences from the Microscopy Society of America, for her many contributions as a technician to the Microscopy Group's successes (Feb. 20, 2013).
- Jim Keiser - received the Engineering Division of TAPPI Award (Technical Association of the Pulp and Paper Industry) (Aug. 6, 2013).
- Jim Keiser - received the Joseph Wolf Founders-Stowe Woodward Award for Best Corrosion and Materials Engineering Paper from the 2012 conference from the Engineering Division of TAPPI (Aug.6, 2013).
- Hua-Tay Lin – selected to receive the Distinguished Engineering Achievement Award from the Engineers' Council, nominated by Mrityunjay Singh of the Ohio Aerospace Institute, NASA Glenn Research Center (2013).
- Hua-Tay Lin - received the 2013 Honor Medal of Aurel Stodola by the Slovak Academy of Sciences, for his research activities and significant contributions in sciences on high temperature mechanical behavior of ceramics and composites (2013).
- Karren L. More – received the 2013 DOE Hydrogen and Fuel Cells Program R&D Award, in recognition of outstanding achievements in leading ORNL's project on Microstructural Characterization of Fuel Cell Materials and in her role as a subcontractor on multiple DOE-funded fuel cell R&D projects (2013).
- Yutai Katoh – was elected as American Ceramic Society (ACerS) Fellow (Mar, 2013).
- Eliot Specht – was elected as American Physical Society (APS) Fellow (2013).
- Amit Shyam – nominated to serve on the Board of Review of the newly formed TMS energy-related journal, Metallurgical and Materials Transactions E. (2013).
- Peter Blau – was invited to be Co-Editor-in-Chief, for the Elsevier journal named "Wear". An international journal on the science and technology of friction, lubrication and wear. He is the fifth EIC, succeeding Prof. Ian Hutchings of Cambridge University (2013).
- T. S. Byun – awarded the distinction of Best Reviewer for 2012 from the Journal of Nuclear Materials, for completing a large number of reviews for the Journal in a timely response and being extremely thorough and constructive for the authors (Apr. 11, 2013).
- Lynn Boatner, Joanne Ramey, Hu Longmire – research featured in the 2013 Allied High Tech Products, Inc. March Calendar in the form of a color micrograph (2013).
- Lynn Boatner, Joanne Ramey – research featured on the cover of the textbook: Essentials of Materials Science and Engineering (3rd Edition) by Donald R. Askeland and Wendelin J. Wright (2013).
- Zhenzhen Yu, postdoc – won the TMS Technical Division Young Professional Poster Contest in Structural Materials Division, titled, "In-Situ Probing of Microscopic Deformation Kinetics in Advanced High-Strength Steels, at the TMS 2013 Annual Meeting & Exhibition in San Antonio, TX (2013).
- Xinghua Yu, postdoc – won the Henry Granjon Prize from International Institute of Welding, in Category B – Materials Behavior and Weldability. His paper was based on his PhD research that explains the fundamental mechanisms of drastic improvement in creep resistance Cr-Mo steel welds (2013).
- Lithium-Sulfur Battery research was featured on the inside front cover of a recent issue of Advanced Functional Materials, by a team of ORNL researchers, Chengdu Liang, Zengcai Liu and Wujun Fu (CNMS); and Zhan Lin and Nancy Dudney (MSTD), co-authors (2013).
- Jim King's farewell tune
ORNL Today, July 1, 2013 - Jim King retired at the end of June. The manager of the Reactor Power Systems Program, which produces the iridium clad vent sets for NASA's deep space exploratory spacecraft and Mars rover generators, got a serenade from Materials S&T Division Director Gene Ice at Thursday's retirement party.
- Critical materials hub gets rolling at ORNL
ORNL Today, June 24, 2013 - Ames Laboratory's Alex King, director of the Critical Materials Institute, helped kick off ORNL's involvement in the new DOE innovation hub last Friday at the JICS auditorium. Associate directors Michelle Buchanan, Martin Keller and Jim Roberto joined King in congratulating ORNL staff for their hard work in preparing for and winning the CMI proposal. Brian Sales will serve as deputy of the hub's "Developing Substitutes" focus area.
- ORNL teams help KARM double Dragon Boat total
ORNL Today, June 24, 2013 - Teams from ORNL fielded, or launched, four Hong Kong-style boats in last Saturday's Knoxville Area Rescue Mission Dragon Boat Races at Concord Park.
- Salsa Picante was a division team lead by the Materials S&T Division, led by Gene Ice.
- The Atoms Family was a Chemical Sciences Division team led by Phil Britt.
- The Dragonettes was an all-woman team led by Machelle Sumner.
- The Titan Rods was a team sponsored by the Early Career Professionals group.
- Competition was fierce and thrilling, as evidenced by Salsa Picante's come-from-behind victory in one heat. The 62-boat race raised more than $250,000 for KARM programs and services, doubling last year's total.
- A Brighter Future for Manufacturing, 3-D Printed 1 Layer at a Time
ORNL in the News (Scientific American), May 8, 2013 - Oak Ridge National Laboratory's robotic prosthesis looks like something out of medieval times—a hand clad in chain mail more appropriate for wielding a broadsword than a mug of coffee.
- TS Byun recognized as reviewer of the year
ORNL Today, April 16, 2013 - The Journal of Nuclear Materials has awarded Thak Sang Byun the distinction of Best Reviewer for 2012. Byun is a research staff member of ORNL's Nuclear Materials Science & Technology Group in the Materials Science and Technology Division. Byun's selection is based on a vote of all the journal's editors. He has completed a large number of exemplary reviews for the Journal that were found to be thorough and constructive for the authors.
- Zhang, Specht elected fellows of American Physical Society
OAK RIDGE, Tenn., April 15, 2013 - Two researchers from the Department of Energy's Oak Ridge National Laboratory have been elected to fellowship in the American Physical Society (APS). Xiaoguang Zhang and Eliot Specht were named APS fellows in recognition of their outstanding contributions to physics.
- RNL lithium-sulfur battery work featured in AFM
Lithium-sulfur battery research by a team of ORNL researchers was featured on the inside front cover of a recent issue of Advanced Functional Materials. Zhan Lin and Nancy Dudneyare the paper's coauthors.
- Valentino Cooper, received the Early Career Research Program Award, by the U.S. DOE Office of Science, for First Principles Predictions of Phase Stability in Complex Oxides., May 7, 2013.
- Wu Zhou, was awarded Best Materials Applications Paper for Year 2012, "Single Atoms Microscopy," by the journal Microscopy & Microanalysis, for the imaging the structure and identifyng the local chemistry within monolayer graphene using ShaRE's Nion UltraSTEM operated at 50kV, May 6, 2013.
- May 2, 2013, Amit Shyam has been nominated as member of the Board of Review for the new TMS journal Metallurgical and Materials Transactions E.
- 3,000 inventions and counting
ORNL Today, Feb. 25, 2013 - Invention Disclosure No. ID-3000, titled, "Rapid Electro-Magnetic Heating of Nozzle in Fused Deposition Modeling," was submitted by Vlastimil Kunc, Orlando Rios, Lonnie Love and Chad Duty on Jan. 23.
- Pulse Thermal Processing, forest monitoring system win FLC awards
ORNL Today, Feb. 25, 2013 - ORNL technologies are among winners of the 2013 national and the southeast region Federal Laboratory Consortium Awards. Awards were given in Excellence of Technology Transfer and for Interagency Partnerships.
- Reeves wins microscopy society's outstanding technologist award
ORNL Today, Feb. 25, 2013 - Shawn Reeves of the Materials S&T Division's Microscopy group has been awarded the 2013 Outstanding Technologist Award in Physical Sciences from the Microscopy Society of America.
- New material promises better solar cells
ORNL in the News (PhysOrg), Feb. 13, 2013 - Researchers at the Vienna University of Technology show that a recently discovered class of materials can be used to create a new kind of solar cell...The team from TU Vienna was assisted by Satoshi Okamoto.
- ZnO grown at ORNL on textbook cover
ORNL Today, Feb. 11, 2013 - The research work of Lynn Boatner and Joanne Ramey of the Materials Science and Technology Division is to be set in stone, or crystal rather, on the cover of the textbook, Essentials of Materials and Engineering (Third Edition), by Donald R. Askeland and Wendelin J. Wright.
- Hua-Tay Lin honored by Engineers' Council
ORNL Today, Jan. 4, 2013 - Hua-Tay Lin has been selected to receive the 2013 Distinguished Engineering Achievement Award from the Engineers' Council.
- Daniel and Preston receive "40 under 40" awards
ORNL Today, Jan. 9, 2013 - ORNL's Claus Daniel was selected by the Greater Knoxville Business Journal as two of 2012's "40 under 40" award winner. The 40 winners were selected "for their passion, commitment, visibility, diversity and overall impact in shaping our region and making it a better place."


text:
Phoenix Arizona SEO Company
We help Arizona small businesses soar to the top of search rankings by crafting thoughtful long term SEO strategies tailored to put our clients ahead of their competition and keep them there!
If the success of your small to medium sized business depends on traffic from search engines, don't rely on DIY SEO guides and bits of search engine marketing news you pick up online. You need real, experienced professionals that you can count on to keep your site on the top of search results!
Our team specializes in helping Phoenix small businesses rank on the first page of Google, Bing and Yahoo for valuable, relevant keywords.
Choose Epica for SEO services because we…
- have 15+ years of successful SEO experience
- have managed dozens of successful local SEO campaigns
- are a preferred Phoenix SEO company
- have an A+ rating / 5 star rating with the Arizona Better Business Bureau (BBB)
- are members of the Arizona Small Business Association (ASBA)
- are members of Local First Arizona small business community
- are experienced, certified SEO professionals
- offer a comprehensive array of search engine optimization services
Get a Free Consultation & discover how expert SEO can help grow your Phoenix business!
Phoenix SEO Consultants
You can count on our team of expert search marketing consultants – we have years of experience crafting & managing successful search campaigns for Arizona businesses just like yours. Our expert SEO services can help you grow your business, get more traffic, and generate more revenue!
Comprehensive SEO Services
We provide a full array of search optimization services from keyword research & page optimization to content development & copy writing. Every step of your campaign is tailored to provide your business the greatest potential for online success!
SEO Strategy
- Insightful market research
- Customer-targeted content strategy
- Link authority development strategy
On Site Optimization
- Rich, in-depth keyword research
- Expert on-site & page optimization
- Compelling, relevant copy writing
Tech & Infrastructure
- SEO audits & technical assessments
- Site template analysis & optimization
- GWT & analytics setup
Off Site Optimization
- Content creation & distribution
- Authority development & link building
- Social media audience engagement
Phoenix SEO Company
We're located in the greater Phoenix, Arizona metropolitan area – with consultants located across the Valley, in Phoenix, Scottsdale, Gilbert & Mesa. Our team of SEO experts work with small & medium sized businesses through-out Arizona to become more visible online through SEO.
Our company is a member of several Phoenix business communities including the Arizona Better Business Bureau (AZ BBB), the Arizona Small Business Association (ASBA) and Local First Arizona. We enjoy working along side our business owner partners all across the state including a local fire station in northern Arizona, a spa in Gilbert, and a bankruptcy lawyer in Phoenix to mention a few.


text:
Moderators: Tealer, Indaswamp, Dep6, Preacher1011, lostpup, La. Hunter, steve-o
ACEBLDRS wrote:Dang it I hate it when you guys post up pics of the pups. Man, it makes me want another sooooo bad.
Good lookin pups. Who's are they? Both yours?
Snafu518 wrote:The flowers smelled of peace and tranquility. Then I burped and got another beer.
No more pups ACE...
but this.....get your duck stamp.
AlaskaRedneK wrote:......And use it to buy that new Over/Under :mrgreen:
Users browsing this forum: sask-hunter and 7 guests


text:
can you fit 19x9.5 in the back of a mkIV jetta without any clearance issues or mods to the fenders? and anyone know what offset if you can? the car is lowered 1.75"
can you fit 19x9.5 in the back of a mkIV jetta without any clearance issues or mods to the fenders? and anyone know what offset if you can? the car is lowered 1.75"
17x8 ET48 Audi A8 Fat Fives on a B5 Passat sedan 1.8t Lowered 1".......Will they fit help please!!
I have a 08 2.5 Jetta and I'm running fk coils I bought Mercedes wheels and they are staggered 18x8.5 and 18x9.5 with et43 I can't figure out what tire size to run I know that most ppl run 215 40 and 225 40 but I would like more tire just because it seems safer. What is the widest tire I can fit? Or would I just be better off running 225s and 215s any info helps thanks
That would make your front tire a lot shorter. You want to keep close to the same outer diameter. That is why you would run a 225/40 and a 245/35. The second number is the ratio of height to width in a percentage; ie 40% of 225 and 35% of 245 for the sidewall.
Got ya so 225 40 and 245 35 would have the same height in the side wall
It will look even?
technically a 255/35 and 225/40 do, but if may run into rubbing problems then a 245/35 is an ok substitute. They will look even. There is only a 1.3% difference in their rolling diameter, and a .17'' difference in sidewall height.
Thanks a lot dude you've been a huge help. One more question is a 225a stretch at a am I going to be able to go low? Would I be better off running a 215 with a 245? Any chance u know anything about lug nuts lol
There is very little about wheels, tires, and suspension that I don't know about.
I'm familiar with lug nuts of many kinds.
225 on a 8.5 is mild stretch (and it depends on the tire as well).
Here is a Bridgestone Potenza 225/40/18 on a 8.5''
Going low is a game of offsets as much as it is tire widths. I really consider people who stretch tires and use smaller tire sizes as cheaters.
Here is how my car sits with 225/40/18 on 8.5 et27f and et23r (and bags)
Last edited by JDriver1.8t; 06-15-2010 at 07:00 PM.
sent pm btw ur car is sick
Hey, haven't been on in awhile but had a quick question but first let me set it up: as i mentioned in previous posts, I came into a set of wheels that had an offset of +48mm. I went ahead and ordered 17.5mm spacers and 14x1.5x45.5mm conical lug bolts. Turns out the wheel holes are too small. The cone shape of the lug bolts won't allow them to sit right. They won't go in all the way.
As I've read in previous posts/threads, my problem has come up with other members.
I was hoping you'd have some insight as to where i might find tuner lug bolts that would solve my dilema? I have found 40mm tuner lug bolts but that's obviously 5.5mm shy of what i was looking for. Would it be unsafe to run them? Thanks
Hello all,
I have a 2008 Eos with stock 16 inch rims and tires. I am interested in a set of OEM 17" by 7" wide rims with 235/55/17 tires off a Tiguan that has an offset of ET43. In researching my rims, they are 16 by 7 with an offset of 45. Is a 2 (mm?) offset an issue? I know the speedo will be off (and I can VAGCOM it) but that is not an issue. And it appears that both rims have a 57.1 mm center bore so that should also fit. I just don't have enough knowledge of how the offset factors in, other than if it is off, the tires may run against the inner fender or the suspension...
And what if the rim width was 7.5 inches? What offset would I need to not have to use spaces or whatever -- just a clean bolt on and go rim?
Thanks for the help!
hi , i have been using the site for a few days now and already got loads of help scanning the pages of the forum. I have now joined and am seeking just a little bit more info if someone doesn't mind. My Mrs has just picked up a peach of a Lupo, or i should probably say a lemon due to its striking yellow.
It currently sits on 13" factory fitted steels. At some point i would like treat her and get some alloys fitted. Without going though any issues of lowering/stiffening or modifying at all can anybody tell me which size rims and tyres i should look for, the only info i got so far was not to exceed a 14" rim otherwise she may need to get stiffer coils/suspension. Thanks for reading.
this is my new wheels
18x8.5 front et44
18x9.5 rear et44
my front fenders are rolled. an have fk coils
going from 5x100 to 5x112 what size adapters for the front? looking to be as flush possible don't mind a lil poke if i can't get flush. thinking 16mm adapters up front they told me thats the smallest they make.
rear 18-20mm adapters is what i was thinking
tires thinking
205/40/18 front
215/40/18 rear
what do u say i should do to make them fit.. more confuse about how will they fit on the front..
In the rear, I would stick to 18 or 20mm as well. You will be flush, to slight poke possibly. I would run a 215 in front and a 235 in the rear for mild stretch. If you go with a 205 in front, then use a 225 in the rear to keep the stretch near the same front and rear.
I have a set of ADR Cyphers that are 18x7.5 with a 48mm offset. what size spacers will i need to get these to a stock stance? and what size spacer would i need to sit flush with the fenders?
To late to call around and it would seem that you would be the person who knows whats up with wheel fitment. I have a MKIII with the standard 15x6.5 ET43 'Bugs and I will be running a 195/50-15 Bridgestone RE-11's. My question is I would like these guys to fit pretty flush and am in need of the correct size spacer to run to achieve this. Thanks for your help
Your MKIV is the portrait of wheel and tire fitment perfection by the way!
I can't get you flush without knowing how low you are, but I can get you close.....
Because they are so narrow, true flush would be had with 30mm front and 40mm rear spacers. I don't recommend going over 25mm without getting the bolt on variety, that bolt to the hub, and then you bolt the wheel to the spacers. Your wheels would still look decent with 20mm front spacers and 25mm rear spacers.


text:
Miley Cyrus Featured in will.i.am's "Feelin Myself" Music Video: Watch Now!
Fresh off her AMA performance, Miley Cyrus stars in the music video for her latest collaboration with will.i.am.
The clip for will's single "Feelin Myself" hit the web on Tuesday (November 26) and also featured Wiz Khalifa and French Montana.
"Feelin Myself" is the first single off of the former Black Eyed Peas crooner's fourth studio album #willpower, which is now available on iTunes.
Watch the new music video below!


text: MBA 10.8.4 MBP 10.6.8 the mba will not read the mbp's library, but the mbp can read the mba's library. both were working perfectly fine until I put the mba to sleep. also, when I added password protection to my wifi connection, the mba would not remember the wifi connection everytime I woke it up. Now that I fixed that, it will not connect to my LAN. it will show up on the sidebar but it will disappear after failing to connect. Then I connect it from Go and it works fine again. everything was working fine until the last update.


text:
i own an e36 323i and a fortnight bk the abs and asc lights came on then when driving they went off on there own then bk on. i have carsoft and all i get is module does not respond. i look it to bmw specialist and his machnine wnt read either or the snap on machnie so i assumed it was abs pump module so fitted another one and still no joy, all the relays are ok the fuses ok no errors on dme the abs and asc are not working i have ran out of options now i have had the dme out all connections are fine all connections to fuse box fine, please canyou give me an idea as to what it is im pulling my hair out! thanks
ihave also tested the asc strangler which chokes the engine when activated thatis working fine


text:
We're here to help!
Log in to see your order history.
Request a return to get started.
For questions on shipping, restocking, and other policies, see our FAQ.
Good Morning Reet – are you on your way to another day at the office? I remember it well. Friday's were always such a delight – knowing that there were two days of blissful freedom coming. Hope your day is a good one.
it is under the designs part…
you are very welcome
Thank you for accepting me!
Find More Beaders »
We send about one e-mail a week with news & deals.We'll keep your information private. Promise!
9 crazy new bead deals every 5 minutes
Deals in your inbox every Tuesday at lunchtime
Picked by our customers for every day savings
We accept the following:
All images, content, and code ©2002-2017 Lima Beads. All rights reserved. See our Privacy Policy.


text: Specifically the Nighthaw. I do realize the battery may need to be replaced in 10 years. What I'm asking is if I was to change the solar battery if it died, how long the watch would last. Is there any chance it would last as long as a mechanical would?


text:
Alonso S. Perales (1898-1960) was a civil rights lawyer and diplomat primarily concerned with Mexican-American civil rights. As such he participated in various Civil Rights organizations and was one of the founders of the League of United Latin American Citizens (LULAC) in 1929. As president of LULAC one of Perales greatest accomplishments was the successful defeat of the Box Immigration Bill in 1930 which would have put a quota on Mexican immigration to the United States. His other civic activities included founding the Independent Voters Association, a Mexican-American political club in San Antonio, introducing legislation calling for the prohibition of racial discrimination, and membership in other religious and civil rights organizations. For his civic activities he was decorated by Spain with the Medal of Civil Merit, one of the nation's highest civil honors.
The collection documents Perales' life and work and contains correspondence, organizational papers from LULAC and other civil rights organizations, publications, and various other items and memorabilia.
The exhibit will run from December 21, 2011 - March 1, 2012 in the M.D. Anderson Library.
Please check out the Perales conference for more information about Alonso Perales.


text: RA'ANANA, Israel, August 13, 2012 /PRNewswire/ -- NICE Systems (NASDAQ: NICE) today announced the introduction of its new release of the NICE Inform multimedia incident information management solution. The new release of NICE Inform builds on NICE's advanced capabilities by giving security control centers and Public Safety Answering Points (PSAPs) the ability to integrate text, video and voice recordings from third party capture platforms into holistic incident timelines. This differentiating technology reinforces NICE's support of Next Generation 9-1-1 and its commitment to meeting the challenge of external data integration for enhancing investigations and debriefings. NICE Inform provides the complete picture for PSAPs, airports, public transportation systems, banks, and other markets that need to synchronize diverse multimedia for incident reconstructions used in investigations and debriefings. This information can also be used to improve future responses by helping organizations identify opportunities for training operators to help improve their skills. The latest release of NICE Inform allows third party recordings from numerous sources to be imported into the NICE Inform organizer in various standard formats and then synchronized with NICE video and voice recordings and other multimedia input. This enables security operations to leverage their existing capture platforms to the fullest, and to reconstruct events exactly as they happened for more thorough investigations. For example, PSAPs can now amalgamate video from citizens' smart devices, in-car mobile video recordings, and video recordings of incidents captured on third party platforms to create comprehensive incident reconstruction timelines. A bank that uses different Video Management Systems (VMS) for ATM transactions and building security can merge video from these different systems for a complete reconstruction of any event. Mass transit systems and airports that manage surveillance video through multiple VMSs can also take advantage of this advanced technology. Using NICE Inform Version 5, they can piece together all elements into one seamless timeline, regardless of the capture platform.


text:
Elena Plotkin
Elena Plotkin Poems
Page :
Page :
Best Poem of Elena Plotkin
I grew up with a bunch of others,
Just me with my sisters and brothers.
When I was younger, I was green and tough,
But as I grew older, I discovered life could be rough.
I changed and grew softer, some called me yellow.
But I was no coward, just a very cautious fellow.
Because where I come from you never know the day,
That your whole family gets eaten or just carted away.
To Be
To be loved by someone other than family,
To be treasured by someone other than friends,
To be respected by someone other than colleagues,
Is to be regarded with envy by both women and men.
(C) 2013 Copyright Elena Plotkin


text:
Intoxicated Driver Crashes Into Family, Kills One
DIXIE COUNTY - A man who got kicked out of a Suwannee restaurant for being belligerent got behind the wheel while intoxicated and caused a crash that took the life of another man. The man that died was the husband of an employee from the restaurant he was just escorted out of. The worker and 5-month-old baby were also part of the wreck.
Makion Kirkland Jr. Was driving his truck while under the influence on County Road 349 in Suwannee. Kirkland crossed the center line and went against traffic, hitting a woman, Sesaley Bretz, and her husband, David Dewees Jr., who were traveling in different cars.
"If you're too drunk to drive take a cab, call a friend because this is really heart breaking," Dan Worley who witnessed the accident said.
Dewees had their 5 month old baby in the back seat he was driving alongside his wife who had just gotten off her shift as a server at this restaurant. He brought the baby to work and was following her back to the house. However one of them would never make it back home.
Dewees was killed on scene while his wife was airlifted to UF Health in critical condition. The baby was also transported to the hospital. A cook at Salt Creek Shellfish was also on his way home from work when he saw what happened, he stopped to help.
"I was able to remove the baby from the car seat and tried to calm it down some... Went and checked on the mother and she was in the other vehicle and she was struck there," Worley said.
While she'd only started working at Salt Creek Shellfish recently, the owners and workers here feel for family as her mother and sister also worked in the restaurant.
"It's always a personal choice. Either you want to take your own life in your hands but you have to think your also taking other peoples' lives in your hands also," Worley added.
Kirkland has been arrested and is in the Dixie county jail for manslaughter, driving under the influence and leaving the scene of a crash. Something Worley, says he saw Kirkland attempting to do. "I know after I helped him off the hood... He went into the woods and it seemed like he was trying to hide," Worley said.
Related Stories
- Single Car Crash In Levy County Kills Child, Injures 5 Others
- Family, friends remember 7 children killed in tragic accident nine years ago today
- Missing UF Student's Family, Not Giving Up
- Three Drivers Hit GPD Patrol Cars, All Charged or Suspected of Drunk Driving
- Alachua Mayor Accused Of Attacking Driver & Using Racial Slurs While Drunk
- Two Women In Clay County Are Shot At In Their Car, One Dies
- RTS Bus And Sheriff's Deputy Car Involved In Crash
- Cattle Car Crash
- Family mourns US army vet killed in car accident on Christmas Day
- Drunk driver crashes car into home