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|Mild Cognitive Impairment: Cerebrospinal Fluid Tau Biomarker Pathologic Levels and Longitudinal Changes in White Matter Integrity.
MedLine Citation:
|PMID: 23151827 Owner: NLM Status: Publisher
|Purpose:To evaluate the relationship between (a) pathologic levels of cerebrospinal fluid (CSF) total tau as an index of the intensity of ongoing neuronal degeneration and (b) longitudinal changes in white matter (WM) integrity in patients with mild cognitive impairment (MCI).Materials and Methods:Participants gave written informed consent, and the Norwegian committee for medical research ethics approved the study. Thirty patients with MCI and nonpathologic CSF total tau levels, nine patients with MCI and pathologic CSF total tau levels, and 16 age-matched healthy control subjects underwent diffusion-tensor imaging at baseline and after a mean follow-up of 2.6 years ± 0.54 (standard deviation), with range of 1.58-3.98 years. The effect of diagnosis (MCI vs no MCI) at baseline and CSF tau levels at fractional anisotropy (FA), mean diffusivity, radial diffusivity (D(R)), and axial diffusivity were tested with tract-based spatial statistics. Differences in WM integrity at baseline and follow-up and change over time were compared among patients with pathologic CSF total tau levels (MCI high tau), patients with normal CSF total tau levels (MCI low tau), and healthy control subjects. Linear mixed-model between-group within-subject analyses were conducted to examine differences in rate of change over time in FA and D(R).Results:Longitudinal analysis of regional WM change revealed significant decrease in FA (P = .038) and increase in D(R) (P = .018) in the MCI high-tau group relative to control subjects. For D(R), the changes were regionally specific to the right cingulum and the right superior and inferior longitudinal fasciculi.Conclusion:Reduction in WM integrity was greater in patients with MCI who had the most intense neuronal degeneration as indexed by using CSF total tau, suggesting that these patients might represent a subgroup of MCI with more intense WM degeneration who are possibly at greater risk of developing Alzheimer disease.© RSNA, 2012.
|Inge K Amlien; Anders M Fjell; Kristine B Walhovd; Per Selnes; Vidar Stenset; Ramune Grambaite; Atle Bjørnerud; Paulina Due-Tønnessen; Anders Skinningsrud; Leif Gjerstad; Ivar Reinvang; Tormod Fladby
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Publication Detail:
|Type: JOURNAL ARTICLE Date: 2012-11-14
Journal Detail:
|Title: Radiology Volume: - ISSN: 1527-1315 ISO Abbreviation: Radiology Publication Date: 2012 Nov
Date Detail:
|Created Date: 2012-11-15 Completed Date: - Revised Date: -
Medline Journal Info:
|Nlm Unique ID: 0401260 Medline TA: Radiology Country: -
Other Details:
|Languages: ENG Pagination: - Citation Subset: -
|Center for the Study of Human Cognition, Department of Psychology, and Department of Physics, University of Oslo, Pb. 1094 Blindern, 0317 Oslo, Norway; Departments of Neuropsychology and Neurosurgery, Oslo University Hospital Ullevål, Oslo, Norway; Department of Neurology and Department of Multidisiplinary Laboratory Medicine and Medical Biochemistry, Akershus University Hospital, Lørenskog, Norway.
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My Nonprofit Reviews
Review for Lam Foundation, Cincinnati, OH, USA
When I was diagnosed with LAM in 2005 I was sent home from the hospital knowing nothing except that I had this incurrable lung disease. Those first couple of years were very lonely and very scary for me. Then I was connected with the LAM foundation. With an entire community of people working towards finding a cure and they connected with with other LAM patients. The LAM foundation has given me comfort, more hope and a new family that understands what I'm going through.
More Feedback
What I've enjoyed the most about my experience with this nonprofit is...
the employees and volunteers and the people that they've connected me with
The kinds of staff and volunteers that I met were...
very professional but at the same time very personable, friendly and just wonderful people
If this organization had 10 million bucks, it could...
probably find a cure for this horrible disease. They could help so many more women with LAM in so many ways
How frequently have you been involved with the organization?
About every week
When was your last experience with this nonprofit?

subset: commoncrawl


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Displaying Powerpoint Presentation on CARDIAC ARREST available to view or download. Download CARDIAC ARREST PPT for free.
Presentation Title : CPR/AED + Cardiac Arrest - Welcome to Provena Health
Presentation Summary : Presence Regional EMS February 2014 BLS CE When using the AED to defibrillate a patient in cardiac arrest, you should: analyze the cardiac rhythm every 60 seconds.
Source : http://www.provena.org/usmc/documents/EMS/PPT/2014%20February%20BLS%20Cardiac%20Arrest%20and%20CPR%2Eppt


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Probably greater than a million lives were already saved by the new-day Dodge Mini Ram bumper. Bumpers are installed to both the back and front ends of your vehicle, and is regarded as among its most basic and essential parts. Your bumpers are supposed to take on damage; thus, they're placed on areas of your automobile that are very possible to get hit full-on. Simply fit all of your Dodge Mini Ram's ends with a sturdy bumper, and you won't suffer the maximum amount of damage, in the eventuality of a crash.
Some Dodge Mini Ram bumpers are even made to tactically cave in on impact-in such a way as to impede huge shock to the automobile and its passengers. In this way, the bumper is actually a sacrificial protection device, dealing with the damage so you as well as your passengers don't have to. For utmost safety and security on the road, you'd best make sure that the Dodge Mini Ram bumper you pay for matches your car; the better the fit, the safer you will be. The bumper ensures that you're getting your money's worth in safety and security each time you're taking your ride out for a spin.
Today, relief can be purchased in the form of an excellent Dodge Mini Ram bumper which you may buy from us at Parts Train; with the cheapest prices you'll see on the Web from dependable brands including Rugged Ridge, Street Scene, and Bolton Premiere, how could you possibly go wrong?
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Use our Part Fit Checker to view parts compatible with your vehicle.


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Writers block: does it exist? Anyone who has ever experienced it will nod their head vigorously and say yes, of course it does. Other writers will smugly say they have never experienced it. There are differing viewpoints on exactly what one should call the phenomenon of when ideas grind to a halt, plots get stuck, and why one just can't seem to move the action or the characters forward. Advice to Writers offers some interesting viewpoints of writers from Toni Morrison to H.G. Wells to Irving Stone on what they consider to be writer's block and what to do about it. Some are very amusing.
Whatever one does or doesn't like to call it, many authors get to a point where they'll do anything except sit down and write their book. Carrie Lofty, a romance writer with a passion for history, has a new take on an old problem: it's called 'fiddling' and when writers don't want to write, they fiddle. Let's hear it from Carrie who has the solution to the writer's habit of 'fiddling!'
Are You a Fiddler?
By Carrie Lofty
I like to think that I've been writing romance since I was 13, but that wouldn't be entirely accurate. Although I finished a romantic screenplay at that age, I wouldn't complete another work of romantic fiction until September of 2006. In the interim, I wrote a series of fantastic, incomplete 100-page attempts. Some didn't even make it to 100 pages. I might finish a few chapters, or a chapter … or a scene.
To say that I hit a wall would also be inaccurate. I've never had writer's block. After all, the ideas were always there. And that was the problem! I would dive headfirst into a new idea only to be distracted, a few days or weeks or chapters later, by a newer and shinier one. My husband threatened to stop listening to my writing discussions if I changed stories again.
This carried on until that fateful summer in 2006 when my husband flew to Virginia for an internship. I stayed behind in Madison, Wisconsin with our two daughters, who were then aged two and three. I wanted desperately to finish a novel so that I, too, could have a career that would take me around the country. And more importantly, I wanted to prove it to myself that I could finish. I couldn't stomach the idea of being a wannabe who always talked a good line, but never actually finished their "some day" novel. What I had been doing was obviously not working. I would write a chapter—then print it out, eviscerate it, revise it, reread it.
In essence, I was a fiddler.
After the initial burst of inspiration wears off and the hard work of actually finishing a 300+ page novel begins, hesitant writers are left with only one recourse: to fiddle. Revising, creating note cards, reading craft books, rereading—all of these can be useful skills and techniques, but only if they are used productively. I was using those same techniques to delay. After all, it looked like I was working!
I believe it stems from fear. The new, bright, shiny, beautiful idea in your head can be an intimidating muse to live up to. While first drafts are messy and unwieldy, the idea in your head looks more like a polished novel or a big-screen movie. The contrast between those two—what you're attempting to create and what you imagine—can be so dissonant and daunting as to cause a fatal case of fiddling. It's fatal to your chances of finishing a book, that's for certain.
How did I break the cycle?
I started with a very reasonable goal. I would write 1000 new words every day. No matter what. Rain, shine, cyclone—no excuses. The first night was the hardest. I wanted to go to bed. I wanted to give up, go back, and mess with what I had already written. But I persevered. After days of practice, and then after weeks of success, those 1000 word slogs began to add up. I finished my first draft in 88 days.
Granted, revising the sucker took four months. I was still learning, and that first draft was truly horrible. But I had done it. I had finished my thought. I had resisted the urge to fiddle. What's more, by the time I typed "the end," the beautiful bright-and-shiny idea that I'd started with didn't glow quite so fiercely. It didn't hold as much appeal. I had moved the flawless "one day" idea out of my head. Now it was a tangible "that day is now" novel. My sense of accomplishment and pride was far outweighed by distress about how many revisions it still required.
So…are you a fiddler?
Do you have so many ideas, all of which you hope to get to "some day"? Do you often start a project, only to abandon it part way through in favor of a newer, better idea? Do you obsess about research and character backgrounds, or reread and revise chapters—all to the detriment of producing new words?
Then I would suggest that you need an intervention. Set yourself a small, reasonable goal to produce new words every day. No matter what. Do not reread what you wrote the day before. Once you've filled up your pages, you can go back and fiddle to your heart's content. And if a new, fantastic idea comes along, use it to motivate yourself into finishing this idea first.
That's my take. I hope you glean wonderful things from it. But just like eating healthy or exercising or quitting smoking, the real impetus to make lasting changes in your writing habits begins with you.
Carrie's witty and observant reflections on life can be found at her website and if you're fascinated by historical romance that takes risks, then check out Carrie's latest book Scoundrel's Kiss (and enjoy!).
Some Top Advice
Alan Rinzler (Consulting Editor) has a great post that suggests perhaps one of the reasons you can't finish that novel is because your plot is not clear. His advice is simple and clear: get your outline right.
Matthew Stibbe (writer in chief at Articulate Marketing) suggests that concentrating on writing will get you over your hurdles and offers 22 ways for staying focused on your writing.
Have you overcome your writer's block? Got any great suggestions? Please let readers know.
A blog discussing all aspects of books and writing, and accepting book reviews and author interviews.
About Me
Fiona Ingram
- I can't remember NOT having a book in my hand. My schoolmates called me a bookworm, and nothing's changed since then. I was brought up on the children's classics because my parents are also avid readers. My earliest story-telling talents came to the fore when, from the age of ten, I entertained my three younger brothers and their friends with serialized tales of children undertaking dangerous and exciting exploits, which they survived through courage and ingenuity (of course!).After three university degrees and a sojourn studying drama in London (The Drama Studio) and mime in Paris (Ecole Jacques le Coq), I returned to South Africa and spent a few years teaching drama, and producing and performing in local community theatre. A move to Johannesburg took me in the direction of writing. The Secret of the Sacred Scarab is my first children's book, and the first in the middle-grade adventure series The Chronicles of the Stone. I have also completed the second book in the series: The Search for the Stone of Excalibur. I have just completed Book Three: The Temple of the Crystal Time-Keeper (set in Central America). I also write animal rescue stories to raise funds for various animal rescue organisations. Please visit http://caladriusbooks.com.
Friday, January 29, 2010
Writer's Block: Imagined or Real?
Posted by Fiona Ingram at 5:44 AM
Labels: Alan Rinzler, authors, Carrie Lofty, characters, creative writing, historical romance, Matthew Stibbe, motivation, plot, writer's block, writing tips
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A man driving a pickup truck with a boat on a trailer died after suffering a medical emergency behind the wheel, according to the Florida Highway Patrol.
State troopers said the man sideswiped another pickup truck in the next lane of Interstate 295 near Commonwealth Avenue on the Westside, then veered off into the treeline.
The man was taken to UF Health Jacksonville, where he died.
The other driver was OK.


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|Ovulation induction in polycystic ovary syndrome: No. 242, May 2010.
MedLine Citation:
|PMID: 20848729 Owner: NLM Status: MEDLINE
|OBJECTIVE: To review current non-pharmacologic and pharmacologic options for ovulation induction in women with polycystic ovary syndrome (PCOS).
OPTIONS: This guideline reviews the evidence for the various options for ovulation induction in PCOS.
OUTCOMES: Ovulation, pregnancy and live birth rates, risks, and side effects are the outcomes of interest.
EVIDENCE: Published literature was retrieved through searches of Medline using appropriate controlled vocabulary and key words. Results were restricted to systematic reviews, randomized control trials/controlled clinical trials, and observational studies. Grey (unpublished) literature was identified through searching the websites of health technology assessment and of health technology assessment-related agencies, clinical practice guideline collections, clinical trial registries, and national and international medical specialty societies.
VALUES: The evidence gathered was reviewed and evaluated by the Reproductive Endocrinology and Infertility Committee of the Society of Obstetricians and Gynaecologists of Canada. The quality of evidence was quantified using the Canadian Task Force on Preventive Health Care.
BENEFITS, HARMS, AND COSTS: Benefits include weight reduction and improvements in ovulation, pregnancy, and live birth rates. Potential harms include medication side effects and multiple pregnancies.
VALIDATION: These guidelines have been reviewed and approved by the Reproductive Endocrinology and Infertility Committee of the SOGC.
|Tannys D R Vause; Anthony P Cheung; Sony Sierra; Paul Claman; James Graham; Jo-Anne Guillemin; Louise Lapensée; Sabrina Steward; Benjamin Chee-Man Wong;
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Publication Detail:
|Type: Journal Article; Practice Guideline; Research Support, Non-U.S. Gov't
Journal Detail:
|Title: International journal of gynaecology and obstetrics: the official organ of the International Federation of Gynaecology and Obstetrics Volume: 111 ISSN: 1879-3479 ISO Abbreviation: Int J Gynaecol Obstet Publication Date: 2010 Oct
Date Detail:
|Created Date: 2010-09-16 Completed Date: 2010-12-31 Revised Date: -
Medline Journal Info:
|Nlm Unique ID: 0210174 Medline TA: Int J Gynaecol Obstet Country: Ireland
Other Details:
|Languages: eng Pagination: 95-100 Citation Subset: IM
Export Citation:
|APA/MLA Format Download EndNote Download BibTex
|MeSH Terms
Aromatase Inhibitors
therapeutic use*
Clomiphene / therapeutic use*
Fertility Agents, Female / therapeutic use*
Fertilization in Vitro
Gonadotropins / therapeutic use*
Hypoglycemic Agents / therapeutic use*
Infertility, Female / drug therapy*, surgery
Metformin / therapeutic use*
Ovary / surgery
Ovulation Induction / methods*
Polycystic Ovary Syndrome / drug therapy*
Randomized Controlled Trials as Topic
Weight Loss
Reg. No./Substance:
|0/Aromatase Inhibitors; 0/Fertility Agents, Female; 0/Gonadotropins; 0/Hypoglycemic Agents; 657-24-9/Metformin; 911-45-5/Clomiphene
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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© All Rights Reserved. Child Protection Policy |
Alvie & Insh Church
Site Map
" Fresh" & Fresh Extra are our discipleship groups for S1-4 & S 5-6 and meets in the garage at 9.30 am on Sunday. We welcome any young person who wants to learn more about how to live with Jesus as their guide. We try to build faith in interactive ways that are relevant to the lives of young people .. For information contact Alistair Appleby 01540651623
We opened Cairngorms Healing Rooms in Aviemore - August 10th, 2011 and we continue to develop that ministry alongside people from other churches


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comments sponsored by
Defamation, Eliot Spitzer, Hotties, Pornography, Privacy
Three years after the Client Number Nine scandal, those involved have moved on to bigger and better things. Well, depending on how you define "bigger and better": Eliot Spitzer landed a gig at CNN, while his former call girl, Ashley Alexandra Dupré, now pens a sex column for the New York Post and was featured on the cover of Playboy. But some people who weren't directly involved have had a harder time moving on, namely a woman named Amber Arpaio.
You may remember her name and perhaps even her driver's license photo from this YouTube video released by "Girls Gone Wild." At the height of the Client Number Nine media frenzy, Joe Francis offered Dupré one million dollars to do a "Girls Gone Wild" magazine shoot and promotional tour. He withdrew that offer when he serendipitously realized he already had footage of Dupré from earlier times in his archive. Dupré then sued him, saying she was only 17 at the time that footage was shot.
Francis responded by releasing a video of Dupré mugging for the camera in a towel, claiming to be 18, and saying her name was Amber Arpaio. The camera then lingers on Arpaio's New Jersey license for about 30 seconds. The video was widely circulated on the Web, and led Dupré to drop her lawsuit — Francis and 'Girls Gone Wild' were triumphant!
Well, until Amber Arpaio filed her own lawsuit against Dupré and "Girls Gone Wild," for defamation and invasion of privacy…
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The Canvas is an arts-based teen center conceived and run entirely by youth for youth.
Promoting values such as social, artistic, and intellectual development and skills relating diversity, privilege, and power, the Canvas uses art to build community and improve the lives of youth in Saint Paul and surrounding areas.
The program provides independent workspaces, workshops and seminars in the arts, and discussions and events on issues important to youth.
Check out out the video
Follow us on Facebook!
Address: 1610 Hubbard Ave (Hancock Rec Center)
Phone: 651-298-4393


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Securing land for the school
The school stands to be in need of its own land to build up classes which can ease the poor conditioned structure that children are learning from at the moment Neighbours can look into this fact and find out possible ways of raising funds to purchase land and build our own school which can later or sooner serve a full primary school.In Kenya a full primary school needs up to eleven rooms.
Hi Jeremy and MariaZ
Happy to hear from you Jeremy.You are most welcome to Mathare village and with your idea contribution,Patmos can go a step.As for the website updates, I asked Marina about it but she was very positive in her response.She simply said that whatever I need concerning it then I should just find out from her.I think you have seen the errors have been rectified and its pages are all highlighting issues concerning patmos.We can now go a head and make it a tool of use for the project.
Thanks all,
Hi MariaZ,
I think it would much easier to update the site if the organization being represented in the site has the full access of the website. Otherwise it would be difficult considering that Marina may have lot of sites to attend to or if she may no longer be available to volunteer.
I was wondering if the access rights can be given much easily or if anyone in the organization being represented can do the editing for her.
All the best,
Jeremy Ecle
Hi neighbors,
I hope each of you is doing well.I suggest that as Shaun is trying other places in getting a grant for the milling machine, we can access other tasks and come up with suggestions a bout them.We still have a problem of writing materials for example.if we can pick that one as a subject of discussion,I think we can get to some means of acquiring them.The question of sponsorship can also be brought to discussion since we have the website online now.How do you find this idea?
Hi Maria,
Marina has now edited the Kilele name on the website.As you had suggested, this will be the tool to draw people's attention to our project and to reach donors, you can go ahead and do as you wanted.I mean creating a task as a means of drawing volunteers and perhaps sponsors for the children.
Hi Richard,
This is why you should also ask her if it is possible that somebody else can update posts on your website, at least you should get the access rights, otherwise the website might soon become outdated. And, yes the website is just a tool to reach donors, the most difficult is to draw people´s attention to your website and your project.
Hi Shaun and Maria,
It is very true that Kilele name is still found on the sponsor the child page.I had reminded Marina to make a change of this unfortunately she couldn't and I sense it can cause a lot of confusion.Let me remind her about it and see if she can edit this name.I hope with the website we can reach sponsors however long it may take.
Hi Richard,
It is a great website but it is really important to get the section under 'Sponsor a Child' and 'Children that need sponsors' changed so that it reads Patmos Junior School and to contact Patmos Junior Foundation. At the moment it says Kilele Junior School and to contact the Kilele Foundation. I agree with Maria that this needs to be edited otherwise it will confuse people. Can you arrange for Marina to do this Richard? Otherwise it is great.
Take care,
Hi Richard,
Great website! If this is fine for you, I could create a task to find volunteers who could search websites and ways on how to promote the child sponsorship scheme. From my experience with John´s school in Komarok, I know that it is very difficult to find sponsors that are not your direct friends or family, but it ´s worth trying.
I was just wondering - do you have actually the rights for the website and the means to edit documents, the child profiles etc by yourself? This is very important since Marina Klimi might not always be available or even disappear as a volunteer. Also, and that is a little bit sad, apparently Marina copy pasted from the website I created for Kilele Junior School in Komarok ... it´s ok to take ideas etc, but you should make sure to modify them and most importantly replace Kilele Junior school by your school name. Could you check this with Marina?
Hi Mary and every one,
Good to learn that we are all looking forward for the grant proposal to be funded.We are not however discouraged by the negative reply from KTCT but we are wishing Shaun a break through in other potential sources that he is getting in touch with.
I want to let you know that Klimi has helped us a lot and now the website is online .I must thank her so much for the efforts and the costs she has put in order to get this website online.Now neighbors can reach other friends by refering them to read more about our project on www.patmosjuniorschool.org.
Thanks to all,
Hi Shaun
That is a shame with Kitchen Table as I know several groups who would be interested in appplying to them. Wish they would update their website to say they are not accepting applications at the moment, it would make life easier for them and potential applicants.
You could check out the list of potential donors at http://docs.google.com/View?docid=dgs9fnp6_0g5zp8p3n which I add to as I find things, and am happy to share with anyone. Also Funds For NGOs website http://www.fundsforngos.org and NGO News Africa http://www.ngonewsafrica.org (which I am not linked with) list latest funding opportunities. Apologies, I don't have the time to look through and find the ones you could try.
Good luck!
Hi Richard and everyone,
I have had a reply from Kitchen Table Charities Trust who have indicated that they have a long waiting list and they are unable to accept any new applications at the present time due to lack of funds. They have said when the situation improves that they will get back in contact with me. This is disappointing but we must now work harder to find other sources of funds.
As Maria suggests,I am attaching another draft of the grant application which I have prepared. I would be grateful for comments and suggestions from neighbours on any way this application can be improved. I will continue to look for donors for this project but the more neighbours who come on board to actively help with sourcing funds, the more chance there will be of success.
Hi Shaun,
Just very quickly on your point on posting the application. The easiest way is to attach it as a document here on a post, and people post comments or do track changes and post it again.
You and your group are amazing, you are really making progress, even it is little by little!
best from sunny Paris,
Hi all,
We have managed to open the account with Kenya Cooperative Bank Kariobangi Branch in Nairobi. We are happy for all the steps we have gone as a result of your advice. This will enable us to make savings for our little contributions and to give easy way to those who will wish to donate towards the work we are undertaking.
We wish Shaun the best in what he is working on i.e the posho mill grant.
God bless you all,
Hi Richard,
Thanks for getting all the information together for the application ; as I mentioned to Maria I shall now get in touch with KTC. I think the only information we don't have is the Bank Account details?
I can uderstand your interest in renting the 4 rooms for the school. At Ksh 10,000 ($133) per month it all comes down to whether you can afford it at present and your priorities. My concerns would be whether you could sustain these payments long term without compromising on other more urgent needs for the children. With the limited funds you have is not the feeding programme and obtaining desks and stationery a higher priority at present?
Just a few initial thoughts. What do other Neighbours think?
Cheers Shaun
Hi Maria,
I shall be getting in touch with Kitchen Table by e mail today to check on the point you raise. I have waited until we are near to being ready to submit an application, which should be next week.
I will copy you in on the final application...ideally it would be good to post it somewhere where everyone can work on it together but I can't see where this is possible on Nabuur.
Keep up your good work with mosquito nets in Uganda. I have some interest on the subject as our other voluntary work is in disaster relief and I have recently designed and made a prototype emergency shelter which I hope one day will be distributed with mosquito nets.
Cheers Shaun


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Telephone (406) 243-6145
Fax (406) 243-4184
Email firstname.lastname@example.com
Borrelia burgdorferi is an etiologic agent of Lyme disease (Barbour and Fish, 1993; Steere, 1994). The bacterium, a member of the spirochete phylum (Baranton and Old, 1995), has a genome predominantly composed of linear DNA molecules (Hinnebusch and Tilly, 1993; Saint Girons et al., 1994). Formulating a medium in which B. burgdorferi grows in vitro was the first step toward a genetic understanding of the physiology and pathogenesis of the organism (Burgdorfer et al., 1982; Barbour, 1984). The growth of B. burgdorferi as single colonies in solid medium (Kurtti et al., 1987; Bundoc and Barbour, 1989; Rosa and Hogan, 1992) has facilitated mutant isolation by selection (Sadziene et al., 1992; Samuels et al., 1994b), although a defined medium for selection of auxotrophs is not currently available. The transformation system described at this web site will be useful for manipulating the spirochete on a molecular genetic level.
Electroporation is the use of an electric pulse to reversibly permeabilize cell membranes (Shigekawa and Dower, 1988) and is an extremely efficient method of genetically transforming bacteria (Nickoloff, 1995). Electrotransformation has been used to disrupt a hemolysin gene (ter Huurne et al., 1992) and flagellar genes (Rosey et al., 1995) in the spirochete Serpulina hyodysenteriae, an etiologic agent of swine dysentery, by homologous recombination. The effect of electroporation buffers and capacitance on the survival of B. burgdorferi has been reported (Sambri and Lovett, 1990). Electroporation has been employed to insert point mutations conferring antibiotic resistance into the gyrB gene of B. burgdorferi by homologous recombination using both PCR products (Samuels et al., 1994a) and oligonucleotides (Samuels and Garon, 1997) as transformation substrates. Electron micrographs of electroporated B. burgdorferi reveal darkly staining regions on the surface of the spirochetes that may represent pores through which DNA can enter the cell (Samuels and Garon, 1997).
Unfortunately, there are both biosafety and physiological limitations on the use of many antibiotics with B. burgdorferi. Despite the use of both kanamycin and chloramphenicol resistance as a genetic marker in S. hyodysenteriae (ter Huurne et al., 1992; Rosey et al., 1995), the successful use of these antibiotics in B. burgdorferi has not been reported and attempts to use them in our laboratory to date have not been fruitful. The coumermycin A1-resistant gyrB gene (Samuels et al., 1994a; Samuels et al., 1994b), currently the only available selectable genetic marker conferring antibiotic resistance in B. burgdorferi, has been developed into an expression cassette that can now be applied toward dissecting the physiology and pathogenesis of B. burgdorferi on a molecular genetic level. This cassette has been successfully inserted into a natural B. burgdorferi circular plasmid (Rosa et al., 1996) and recently used to disrupt a gene encoding an outer surface protein (Tilly et al., 1997 1997 ), demonstrating that the technology is feasible. However, recombination into the chromosomal gyrB locus generates a large background and only about 0.4% of the coumermycin A1-resistant transformants contain the targeted insertion (Tilly et al., 1997 1997 ; Rosa et al., 1996). This requires extensive screening, which is facilitated by picking colonies with a sterile toothpick directly to a PCR tube and using a 96-well thermal cycler (Rosa et al., 1996).
This web site, derived from a previously published book chapter (Samuels, 1995), provides detailed methods for introducing DNA into B. burgdorferi by electroporation and for the selection of transformants (or spontaneous mutants) on solid medium. We typically obtain transformation efficiencies of 103 to 104 transformants/ug of linear DNA. The protocol may work for other species of the genus Borrelia.
Please do not hesitate to contact me at email@example.com with questions, comments or reprint requests.
Although we routinely use high-passage B. burgdorferi strain B31 for genetic experiments, we have had success transforming B. burgdorferi strains HB19 and N40, although we have not confirmed the N40 transformants by sequence analysis. We have transformed low-passage infectious B31, but have not yet assayed the infectivity of the transformants.
BSK (Barbour-Stoenner-Kelly) medium can be purchased from Sigma, but it is expensive. BSK contains 8% (v/v) 10x CMRL-1066 (without l-glutamine and sodium bicarbonate; Life Technologies, Gaithersburg, MD), 4 g/L Neopeptone (Difco, Detroit, MI), 40 g/L bovine serum albumin (BSA; fraction V, Pentex; Miles, Kankakee, IL), 1.6 g/L Yeastolate (TC; Difco), 4.8 g/L N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid (HEPES), 4 g/L glucose, 0.56 g/L sodium citrate, 0.64 g/L sodium pyruvate, 0.32 g/L N-acetyl-d-glucosamine, 1.76 g/L sodium bicarbonate, and 6.6% rabbit serum (trace hemolyzed; Pel-Freez, Rogers, AR). Adjust to pH 7.6 with 1 N NaOH, stir slowly for 2-3 h, and sterilize by filtration (successively through a prefilter, a 1.2 ĉm filter, a 0.45 ĉm filter, and a 0.22 ĉm filter). BSK-H from Sigma comes frozen, but then is stored at 4ĝC for up to 2 months (see Note 1).
dPBS (Dulbecco's phosphate buffered saline) contains 8 g/L NaCl, 0.2 g/L KCl, 1.15 g/L Na2HPO4, and 0.2 g/L KH2PO4. Sterilize by filtration and store at 4ĝC.
dPBS++ (dPBS with divalent cations) is made by adding 0.1 g/L CaCl2 and 0.2 g/L MgCl2.6H2O to dPBS.
EPS (electroporation solution) is 93 g/L sucrose and 15% (v/v) glycerol. Sterilize by filtration and store at 4ĝC.
P-BSK (plating-BSK, also known as 1.5x BSK) medium: add 83 g BSA, 8.3 g Neopeptone, 10 g HEPES, 1.2 g sodium citrate, 8.3 g glucose, 1.3 g sodium pyruvate, 0.7 g N-acetyl-d-glucosamine, 3.7 g sodium bicarbonate, and 4.2 g Yeastolate to 1 L of water (18 Mêcm). Adjust to pH 7.5 with 1 N NaOH, stir slowly for 2 to 3 h, and sterilize by filtration (successively through a prefilter and a 0.22 ĉm filter). Store at -20ĝC.
Coumermycin A1 (Sigma, St. Louis, MO) at 50 mg/mL in dimethyl sulfoxide (DMSO) (see Note 2). Dilute to 10 mg/mL or other convenient concentration in DMSO for a working solution. Store at -20ĝC. We currently use a BTX TransPorator Plus for electroporations, but we have successfully used a BioRad Gene Pulser with Pulse Controller.
Preparation of Competent Cells
1.Inoculate 500 mL of BSK medium in a 500 mL screw-top bottle with 1 mL of a late-log phase culture (see Note 3). Incubate at about 32ĝC (without agitation) until the culture reaches a density of about 5 x 107 cells/mL (see Note 4). This requires 36-60 h.
2.Transfer culture to two sterile 250 mL screw-top centrifuge bottles and cap.
3.Centrifuge at 4000 x g for 20 min at 4ĝC. Decant the supernatant fraction and resuspend each cell pellet in 30 mL of cold dPBS (see Note 5).
4.Transfer cells to two sterile 50 mL screw-top centrifuge tubes and cap.
5.Centrifuge at 3000 x g for 10 min at 4ĝC. Decant the supernatant fraction and resuspend each cell pellet in 30 mL of cold dPBS.
6.Centrifuge at 3000 x g for 10 min at 4ĝC. Decant the supernatant fraction and resuspend each cell pellet in 10 mL of cold EPS. (EPS wash 1.)
7.Transfer cells to two sterile 14 mL polypropylene tubes and cap.
8.Centrifuge at 2000 x g for 10 min at 4ĝC. Decant the supernatant fraction and resuspend each cell pellet in 10 mL of cold EPS. (EPS wash 2.)
9.Centrifuge at 2000 x g for 10 min at 4ĝC. Decant the supernatant fraction and resuspend each cell pellet in 10 mL of cold EPS. (EPS wash 3.)
10.Centrifuge at 2000 x g for 10 min at 4ĝC. Decant the supernatant fraction and pool the cell pellets in 0.6 mL of cold EPS (see Note 6).
11.Distribute 50 ĉl aliquot fractions of the cell suspension into sterile
1.7 mL tubes on ice (see Notes 7 and 8).
12.Unused competent cells can be transferred directly into a -70 to -80ĝC freezer and then thawed on ice 10 to 30 min prior to electroporation (see Note 8).
1.Cool electroporation cuvettes (0.2 cm electrode gap) to 4ĝC.
2.Transfer 1 to 5 uL of a solution containing 0.3 to 1 ug of DNA in water (see Notes 9 and 10) to the cell suspension, mix gently, and incubate on ice for about 1 min.
3.Transfer the cell/DNA mixture to a chilled electroporation cuvette. Cap the cuvette and shake the cell/DNA mixture to the bottom of the cuvette with a flick of the wrist so that the mixture spans the two electrodes.
4.Place the cuvette in the pulse generator and deliver a single exponential decay pulse of 2.5 kV, 25 ĉF, and 200 ê. This should produce a time constant of 4 to 5 msec (see Notes 10 and 11).
5.Immediately (within 1 min) add 1 mL of BSK medium (at room temperature) without antibiotics and mix the cell suspension by pipeting up and down.
6.Transfer the entire mixture to a sterile 14 mL tube that contains an additional 9 mL of BSK medium (at room temperature) and incubate (without agitation) at about 32ĝC for about 18 h.
Selection of Transformants
You will need 35 mL of molten medium for each plate and at least two plates for each transformation. This recipe gives a final volume of 510 mL. 1.Mix 240 mL of P-BSK medium, 38 mL of 10x CMRL-1066, and 12 mL of rabbit serum. Equilibrate the mixture at 55ĝC in a water bath. Autoclave 200 mL of 1.7% agarose (high strength analytical grade), equilibrate to 55ĝC, and combine with the medium mixture. Add 20 mL of fresh 5% sodium bicarbonate and the selective agent (see Notes 2 and 12).
2.Transfer 15 mL of the molten medium into 12 to 14 100 mm dishes and allow to solidify at room temperature. Equilibrate the remainder of the molten medium at 42ĝC.
3.Transfer 0.1 mL of BSK II medium containing the electroporated cells to a 50 mL tube. Add 20 mL of the molten medium (at 42ĝC) and mix by pipeting up and down once. Transfer the mixture to the plates containing the solidified bottom agarose medium and allow to solidify at room temperature.
4.Centrifuge the remaining 9.9 mL of culture at 8000 x g for 5 min, resuspend in 1 mL of supernatant fraction, and plate as above.
5.Incubate the plates at 32 to 34ĝC in a humidified 1 to 5% CO2 atmosphere. Colonies should appear in about 14 days.
6.Isolate single colonies by picking with a plugged 15 cm pasteur pipet (with bulb). Transfer to 10 mL of BSK II in the presence of antibiotics. Cultures should reach late-log phase in 6-9 days.
Screening of Transformants
1.Number colonies using a felt-tip marker on the bottom of the plate. Set up PCR using primers that flank the directed insertion site (see Note 13).
2.Pick colonies with a sterile toothpick and transfer directly into tubes containing a PCR mix (Taq DNA polymerase, buffer, dNTPs, and primers) (see Note 14). Plates are then returned to the incubator for 1 week to allow for expansion of picked colonies.
3.PCR conditions are typically 30 cycles of 94ĝC for 0.5 min, 45 to 50ĝC for 0.5 min, and 68ĝC for 2.5 min.
4.An equal volume (20 uL) of 2x gel-loading buffer is added to each reaction and 20 uL is analyzed by (1%) agarose gel electrophoresis (using a 30-well comb) and ethidium bromide staining. Colonies exhibiting the correct phenotype are picked as described above after 1 week of expansion.
1.The quality of BSA varies by source and lot. We have found Miles to be a reliable source. However, we reserve 5 or 10 kg batches and test samples for the ability to support the growth of B. burgdorferi. We currently purchase BSK-H containing rabbit serum from Sigma. We have stored BSK-H medium (without serum) at 4ĝC for up to 2 years and found that it can support the growth of highly passaged strain B31 upon the addition of fresh serum.
2.The only antibiotic that is not clinically useful and has been shown to be effective for selection of resistant mutants is coumermycin A1 (Samuels and Garon, 1993; Samuels et al., 1994b) However, bactericidal antibodies have been used to select for mutants of B. burgdorferi (Cinco, 1992; Coleman et al., 1992; Sadziene et al., 1992). Novobiocin, another coumarin antibiotic, which is 20-fold less expensive than coumermycin A1 per use, can be used at 5 ug/mL to select coumermycin A1-resistant transformants. Novobiocin is labile upon exposure to light or freeze-thaw cycles; we prepare it fresh.
3.B. burgdorferi is a class 2 human pathogen and therefore should be handled in a class II biological safety cabinet (laminar flow hood). In addition, BSK medium is rich and all procedures should be performed aseptically. Introduction of recombinant DNA into a class 2 pathogen requires permission from the Institutional Biosafety Committee before initiation of the experiments according to Section IIIB of the Guidelines for Research Involving Recombinant DNA Molecules (Federal Register).
4.The cell density (or growth phase) is a significant factor for successful electrotransformation, as is the case with other bacteria (Shigekawa and Dower, 1988; Dower et al., 1992; Nickoloff, 1995). The cells will either not transform efficiently or not plate efficiently if the cell density is too high (when the color of the medium changes). We have had success electrotransforming cultures harvested at 1 to 7 x 107 cells/mL, although a low cell density (1 to 2 x 107 cells/mL) requires pelleting the cells at a higher g force (up to 5000 x g) and adjusting the final volume of the cell suspension (see Note 6). Alternatively, higher cell densities can be used for preparing competent cells but then 5% or less of the transformation should be plated (Rosa et al., 1996). Cell density can be determined using a Petroff Hausser Counting Chamber (Hausser Scientific Partnership, Horsham, PA). Dilute 0.1 mL of the culture with 0.9 mL of cold dPBS++ and place in the counting chamber. Count cells over all 25 groups of 16 small squares in all planes using a dark-field microscope. Multiply the number of cells counted by 5 x 105 to calculate cells/mL. Alternatively, cell density can be determined by spectrophotometry. Centrifuge 10 mL of the culture at 5000 x g for 10 min. Decant the supernatant fraction and resuspend the cell pellet in 1 mL of dPBS++. Centrifuge at 8000 x g for 5 min. Decant the supernatant fraction, resuspend the cell pellet in 1 mL of dPBS++, and measure the A600. Multiply the A600 by 1.4 x 108 to calculate cells/mL in the culture.
5.Thorough washing is important to remove components of the medium (see Note 10). Cell pellets are resuspended in both dPBS and EPS by pipeting followed by vortex mixing. These treatments do not appear to affect cell viability. The two dPBS washes during the preparation of competent cells are not required for successful transformation, although the cells are harder to pellet in EPS without these washes. We have not quantitatively assessed the effect of omitting the dPBS washes, and we still routinely perform them.
6.The final cell concentration should be 1 to 5 x 1010 cells/mL (with a final volume of about 0.9 mL). The volume of EPS used to resuspend the final cell pellet may have to be adjusted to account for initial cell number and efficiency of decanting.
7.We find that use of presterilized aerosol-resistant pipet tips (with aerosol barriers) helps to maintain sterility when handling small volumes of liquid.
8.We have not examined the effect of temperature on transformation efficiency, but maintaining the competent cells at 4ĝC is generally considered to yield optimal efficiencies (Shigekawa and Dower, 1988; Dower et al., 1992; Nickoloff, 1995). Frozen competent cells have a transformation efficiency only about 15% lower than that of fresh competent cells, although we still prefer to use fresh cells whenever possible.
9.We routinely obtain 1000 or more transformants/ug of DNA with strain B31, although we have only used linear DNA generated by PCR as an electrotransformation substrate. Linear molecules are 1000-fold less efficient in electrotransformation of Escherichia coli than circular molecules (Shigekawa and Dower, 1988) and we are currently constructing circular replicons from broad-host-range vectors and natural spirochete plasmids for use in B. burgdorferi.
10.Electroporation in the presence of high ionic strength solutions causes arcing (and a lowered time constant). Two arcs will kill all of the B. burgdorferi cells. We use various commercial DNA purification kits or we ethanol precipitate the DNA and either elute or resuspend the DNA at a high concentration in water. Transformation efficiency generally increases with DNA concentration (Shigekawa and Dower, 1988; Dower et al., 1992; Nickoloff, 1995).
11.Preliminary studies suggest that one pulse effected higher transformation efficiencies than multiple pulses and that varying the resistance from 100 to 400 ê affected the time constant, but did not significantly alter the transformation efficiency. The BTX TransPorator Plus has only a variable voltage setting.
12.An antibiotic concentration that inhibits bacterial growth in liquid culture by 80 to 90% relative to growth in the absence of antibiotics has been used to select for spontaneous mutants and transformants in solid medium (Sambri and Lovett, 1990; Samuels and Garon, 1993; Samuels et al., 1994a; Samuels et al., 1994b; Samuels and Garon, 1997). We currently use 0.2 ug/mL coumermycin A1 for selection of single mutant gyrB genes and 0.5 ug/mL for selection of the triple mutant NGR gyrB gene.
13.PCR primers should be designed so that a negative result (recombination into the chromosomal gyrB locus) yields an amplification product of about 100 to 500 bp and a positive result (directed insertion and gene disruption) yields a product of 2.2 to 2.6 kb (which includes the 2.1 kb coumermycin A1-resistant cassette). Use of a 96-well thermal cycler and 20 uL reactions allows for large-scale screening and saves on reagents.
14.The transfer only requires sticking the numbered colonies with the toothpick and sticking the toothpick briefly into the PCR mix. Use a fresh toothpick for each colony.
I sincerely thank Claude Garon for support during my postdoctoral fellowship when this transformation protocol was first developed, Kit Tilly for genetic counseling, Patti Rosa for the plating protocol and the PCR screening protocol, and Iain Old for making this web site a reality.
Baranton, G., and Old, I.G. (1995) The Spirochaetes: a different way of life. Bull Inst Pasteur 93: 63-95.
Barbour, A.G. (1984) Isolation and cultivation of Lyme disease spirochetes. Yale J Biol Med 57: 521-525.
Barbour, A.G., and Fish, D. (1993) The biological and social phenomenon of Lyme disease. Science 260: 1610-1616.
Bundoc, V.G., and Barbour, A.G. (1989) Clonal polymorphisms of outer membrane protein OspB of Borrelia burgdorferi. Infect Immun 57: 2733-2741.
Burgdorfer, W., Barbour, A.G., Hayes, S.F., Benach, J.L., Grunwaldt, E., and Davis, J.P. (1982) Lyme disease--a tick-borne spirochetosis? Science 216: 1317-1319.
Cinco, M. (1992) Selection of a Borrelia burgdorferi antigenic variant by cultivation in the presence of increasing amounts of homologous immune serum. FEMS Microbiol Lett 92: 15-18.
Coleman, J.L., Rogers, R.C., and Benach, J.L. (1992) Selection of an escape variant of Borrelia burgdorferi by use of bactericidal monoclonal antibodies to OspB. Infect Immun 60: 3098-3104.
Dower, W.J., Chassy, B.M., Trevors, J.T., and Blaschek, H.P. (1992). Protocols for the transformation of bacteria by electroporation. Guide to electroporation and electrofusion. San Diego, Academic Press. 485-499.
Hinnebusch, J., and Tilly, K. (1993) Linear plasmids and chromosomes in bacteria. Mol Microbiol 10: 917-922.
Kurtti, T.J., Munderloh, U.G., Johnson, R.C., and Ahlstrand, G.G. (1987) Colony formation and morphology in Borrelia burgdorferi. J Clin Microbiol 25: 2054-2058.
Nickoloff, J.A., Ed. (1995). Electroporation Protocols for Microorganisms. Methods in Molecular Biology. Totowa, New Jersey, Humana Press.
Rosa, P., Samuels, D.S., Hogan, D., Stevenson, B., Casjens, S., and Tilly, K. (1996) Directed insertion of a selectable marker into acircular plasmid of Borrelia burgdorferi. J Bacteriol 178: 5946-5953.
Rosa, P.A., and Hogan, D.M. (1992). Colony formation by Borrelia burgdorferi in solid medium: clonal analysis of osp locus variants. First International Conference on Tick-Borne Pathogens at the Host-Vector Interface: An Agenda for Research. St. Paul, University of Minnesota. 95-103.
Rosey, E.L., Kennedy, M.J., Petrella, D.K., Ulrich, R.G., and Yancey, R.J., Jr (1995) Inactivation of Serpulina hyodysenteriae flaA1 and flaB1 periplasmic flagellar genes by electroporation-mediated allelic exchange. J Bacteriol 177: 5959-5970.
Sadziene, A., Rosa, P.A., Thompson, P.A., Hogan, D.M., and Barbour, A.G. (1992) Antibody-resistant mutants of Borrelia burgdorferi: in vitro selection and characterization. J Exp Med 176: 799-809.
Saint Girons, I., Old, I.G., and Davidson, B.E. (1994) Molecular biology of the Borrelia, bacteria with linear replicons. Microbiology 140: 1803-1816.
Sambri, V., and Lovett, M.A. (1990) Survival of Borrelia burgdorferi in different electroporation buffers. Microbiologica 13: 79-83.
Samuels, D.S. (1995). Electrotransformation of the spirochete Borrelia burgdorferi. Electroporation Protocols for Microorganisms. Totowa, New Jersey, Humana Press. 253-259.
Samuels, D.S., and Garon, C.F. (1993) Coumermycin A1 inhibits growth and induces relaxation of supercoiled plasmids in Borrelia burgdorferi, the Lyme disease agent. Antimicrob Agents Chemother 37: 46-50.
Samuels, D.S., and Garon, C.F. (1997) Oligonucleotide-mediated genetic transformation of Borrelia burgdorferi. Microbiology 143: 519-522.
Samuels, D.S., Mach, K.E., and Garon, C.F. (1994a) Genetic transformation of the Lyme disease agent Borrelia burgdorferi with coumarin-resistant gyrB. J Bacteriol 176: 6045-6049.
Samuels, D.S., Marconi, R.T., Huang, W.M., and Garon, C.F. (1994b) gyrB mutations in coumermycin A1-resistant Borrelia burgdorferi. J Bacteriol 176: 3072-3075. Shigekawa, K., and Dower, W.J. (1988) Electroporation of eukaryotes and prokaryotes: a general approach to the introduction of macromolecules into cells. BioTechniques 6: 742-751.
Steere, A.C. (1994) Lyme disease: a growing threat to urban populations. Proc Natl Acad Sci U S A 91: 2378-2383.
ter Huurne, A.A.H.M., van Houten, M., Muir, S., Kusters, J.G., van der Zeijst, B.A.M., and Gaastra, W. (1992) Inactivation of a Serpula (Treponema) hyodysenteriae hemolysin gene by homologous recombination: importance of this hemolysin in pathogenesis of S. hyodysenteriae in mice. FEMS Microbiol Lett 92: 109-114.
Tilly, K., Casjens, S., Stevenson, B., Bono, J., Samuels, D.S., Hogan, D. and Rosa, P. (1997) The Borrelia burgdorferi circular plasmid cp26: conservation of plasmid structure and targeted inactivation of the ospC gene. Mol. Microbiol. 25:361-373.


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As I type this, Barack Obama is on television denouncing his 'old uncle', his 20 year spiritual adviser and mentor. Seems Obama's political advisers got through to him, advised him his campaign was in the crapper thanks to Rev. Jeremiah Wright, and he must throw Wright under the bus (along with his grandmother and mother, both of whom he has previously thrown under the proverbial bus). The Leftist media has made every effort to twist themselves into rhetorical and intellectual pretzels trying to downplay or dismiss, their candidate's monumentally poor judgment in hanging around with these folks (Rev. Jeremiah Wright, Tony Rezko, and William Ayers) for much of his adult life.
I received this email on Barack Obama from someone named Nagol Kluth
I want you to know that for lying and intentionally spreading falsities, you will be judged. It is not within my right to say what judgement will be passed, since that is His domain alone, but you should think twice before knowingly spreading lies.
A Concerned, Educated Reader
I did not answer the email and have no intention of answering. But I will comment here:
Nagol Kluth (if that's your real name), I do not lie, I do not knowingly or intentionally spread 'falsities', and I always think twice before printing anything here at Right Truth.
Additionally I don't claim to be perfect, but I willingly submit to "His" judgment, the God of the Holy Bible, the God of Christians and Jews.
On Barack Obama, he is a 'fake', he is the liar, he is a creation of the Chicago political machine.
... Obama is a bald faced liar. And it would appear just looking at what Tom Blumer has been able to come up with that either Obama looked the American people in the eye during his speech in Philadelphia and lied through his teeth ...
Blumer, by the way, makes a compelling case that Obama was wide awake during the services, went so far as to take notes of Wright's sermons in the space provided by the church bulletin, and purchased at least one tape of Wright's talks.
It's clear who the liar is. It is the fault of some Americans for not being informed, for not searching for the truth, even if it hurts your chosen candidate. So to readers like Nagol Kluth, I say pull the scales from your eyes and search for the truth, even if it hurts.
Other reading:
Rev. Wright's Press Club Debacle Has CNN Anchor Groaning 'Ah, Boy'
Rev. Wright: The Gift that Keeps On Giving
Trackposted to Outside the Beltway, Rosemary's Thoughts, Maggie's Notebook, Adam's Blog, Shadowscope, The Amboy Times, Democrat=Socialist, Conservative Cat, Adeline and Hazel, Faultline USA, third world county, McCain Blogs, DragonLady's World, The World According to Carl, Blue Star Chronicles, Pirate's Cove, The Pink Flamingo, CORSARI D'ITALIA, , Dumb Ox Daily News, Right Voices, The Yankee Sailor, OTB Sports, and Gone Hollywood, thanks to Linkfest Haven Deluxe.


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Civil Rights Litigation Drafting
Based on a hypothetical fair housing case, this course is designed to teach students how to draft documents common to civil rights cases. The course is of special interest to students who are contemplating a career in civil rights law, but it also trains students in general civil litigation drafting skills. Students draft pleadings, discovery requests and responses, letters to clients, settlement agreements, and summary judgment motion papers. This offering differs from Civil Litigation Drafting only in being more specialized in its substantive focus; students may not take both courses for credit.Credits: 2 or 3
Type: SEM
Does this course satisfy the skills requirement? Yes


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The Magazine
California, Too!
Another judicial setback for same-sex marriage advocates.
Oct 23, 2006, Vol. 12, No. 06 • By DAVID M. WAGNER
THE FUTURE ain't what it used to be. You can't count on the inevitable any more. When the highest court in Massachusetts announced on November 18, 2003, that a right to same-sex marriage was contained in the Bay State's 226-year-old constitution, it seemed as if that court's self-concept--boldly leading a retrograde nation into a brave new world--might pan out.
Almost three years later, precisely zero state courts of last resort have agreed. The high court of that electric-blue state New York has strongly disagreed. The latest court to weigh in (at 131 pages) is an appeals court in that other blue-state paradise, California, which on October 5 reversed a trial court in four cases combined under the name In re: Marriage Cases.
The California court, like the one in New York, holds that a decision to recognize same-sex marriage must rest with the legislature and/or the referendum process--in a word, with the people. In the meantime, the Golden State's referendum-passed law barring same-sex marriage--Prop. 22, which passed by a 61-39 margin in 2000--is not irrational, and so must be upheld. (Full disclosure: One of my former students represented the appellant in the California case, and another represented an amicus curiae.)
In a long and somewhat repetitive opinion for the 2-1 majority, Presiding Justice William R. McGuiness hits the major points that are familiar from the New York opinion (and the Massachusetts dissents): Classifications based on sexual orientation get "rational basis" scrutiny, not "strict scrutiny"; California's marriage law is not discriminatory because it affects both sexes equally and is not part of a system of either male or female superiority; the "rational basis" test is deferential to legislators (or voters in a referendum); finally, and above all, major cultural transformations (as even the Massachusetts court admitted it was carrying out) should be the work of the people, not judges.
In addition, the court notes a tension between two of the arguments from the same-sex marriage advocates: the right of privacy, and the right of expression. First, it says, the right of privacy is not an infinitely malleable vessel: It has legal contours, and those do not yet include same-sex marriage. As for the right of expression--which is inherently public, and therefore sits awkwardly beside an argument based on privacy--California's law restricting marriage to a man and a woman forbids no expressive act. It only denies state assistance in one particular act--getting married--that is less an act of personal self-expression than one of self-joining to a communal paradigm.
A pervasive presence in this opinion is the Domestic Partnership Act California first adopted in 1999--i.e., civil unions. The advocates of this measure no doubt thought themselves properly progressive, but to the respondents and the dissenting judge here, they were actually retrograde: They gave same-sex couples every legal right except marriage, and thereby created--wait for it--a system of separate but equal.
Um, no, says the majority. California is in the forefront of giving same-sex couples legal recognition. The state's Domestic Partnership Act in fact renders untenable any charge that California law as a whole is bigoted against gays.
It would be a mistake to see In re: Marriage Cases, even if upheld (it is being appealed to the California Supreme Court), as a ringing affirmation of society's unique interests in traditional marriage. Though it alludes to the theory that the purpose of marriage is to encourage stability in those relationships that are most likely to produce children--and that are uniquely capable of producing unplanned children--the court does not endorse this view, because California attorney general Bill Lockyer disavowed it in presenting his case. Other dicta in the opinion suggest that, on a full trial record with adequate expert testimony (not present here), this court might be willing to hold that homosexuals do, after all, fit the criteria of a "suspect classification"--which would mean "strict scrutiny" (and probable overturning) for any state classifications based on sexual orientation.
The dissent by Judge J. Anthony Kline is a long exercise in a familiar methodology: generality inflation. The Supreme Courts of both the United States and California have held that their respective constitutions contain a "right to privacy," haven't they? Of course they have. And privacy shouldn't be confined merely to the things the courts have previously applied them to, should it? It should be generalized, so as to include anything that contributes to individual self-fulfillment, right? Well, maybe so, in a late-night college bull-session sort of way.


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Truett-McConnell alumna Jessie White, class of '58, was a member of a homecoming court, a cheerleader, a choir member and soloist. She is a parent of a TM alum, and Jessie later served as a trustee for her alma mater. In addition, for 11 years she served in the Truett-McConnell advancement office and was director of alumni relations.
However, September 27 dawned as no other day for Jessie when she awoke in heaven with the Lord she served for her entire life. Jessie's inoperable condition on Earth was healed the moment she went home.
"We are so pleased to have enjoyed a recent visit with Jessie," said John Yarbrough, director of alumni relations. "And I am grateful that your Alumni Association Executive Committee acted rapidly to confer upon such a distinguished alumna the Meritorious Service Award on Sept. 18, just nine day before her passing."
School officials also named the Jessie White Scholarship Fund in her honor.
Dr. Emir Caner, president of Truett-McConnell joined Yarbrough and others on the visit with Jessie.
"How are your children?" Jessie asked Caner when he entered the room.
After Yarbrough showed Jessie some photographs from her 1958 class, he gave her a card signed with well wishes from many who served with her on TM's staff.
"Jessie and I got to know one another when she was director of alumni relations in the 80s," Yarbrough said. "It was a privilege to serve with her at that time as an alum, and it was a privilege to present to her the much deserved Meritorious Service Award."
Jessie was both visibly moved and gracious when receiving the award.
When Caner informed Jessie of the scholarship fund named in her honor, he asked, "What do you want those who receive the scholarship to know?"
"To serve the Lord," she quipped without hesitation. Jessie continued to show her love for her alma mater by sharing the names of people who could assist with the advancement of the college.
A conversation including comments and memories about Truett-McConnell College continued, and then Jessie's pastor, Dan Rosser (Pleasant Grove BC, Ellijay), led the group in prayer.
"Doctors had told Jessie that there was nothing more they could do," Yarbrough told TMNews. "However, Jessie's faith, perseverance and supportive group of family, friends and a loving church fellowship sustained her until she went to be with the Lord."
Others visiting Jessie were here sister, Betty Jean Nealey, and friend, TM alumna Betty Jo Holt ('58).
Jessie's home-going was celebrated Sept. 29 at her church, the Pleasant Grove Baptist Church in Ellijay.


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Comet ISON - The Final Countdown
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Comet c/2012 s1 ISON is just 71 hours away from entering our inner solar system and 'buzzing the towers' on Planet Mars before she swings by to blow Earth, Venus and Mercury a dusty kiss. ISON is a Hyperbolic Celestial Body and she's hot to trot. Ison Has been traveling from 10,000 to Billion years just to do one dance with our Sun and then leave the Solar System. All Governments and All Profffesional scientists swear 100% that Comet ISON has 0% of harming any life on Earth in anyway ever. So that's comforting right? Yeah so. In 12 days, on October 1st, Ison will appear in the field of view of NASA's SECCHI STEREO Ahead camera. That's exciting. But wait! It gets better! Comet ISON will then be in the field of view of decent SC SPACE NAVY / NASA / GSESAUN cameras until the middle of January 2014*! We've got BRRISON ! Balloon Rapid Response for ISON! It's a a special stratospheric balloon that floats right above the top of Earth's atmosphere and it will haul around an awesome Robot/Computer/Camera/Sensor! will observe Comet ISON in the near-infrared and in the near-ultraviolet and visible wavelength ranges at an altitude of 120,000 feet. The near infrared camera will measure the ratio of carbon dioxide (CO2) to water (H2O) emissions from the cometary nuclei as a vital diagnostic of the comet's origins. Oh. And, the last week broke Meteor, Meteorites and Fireball records. I even saw a giant yellow & green one over Texas. It came from the South East. It was huge, bright and lasted about 7 seconds. That's what she said???. ugh. yeah. And, Our Government is about to shut down. So if NASA got hit very hard by the sequester, I imagine the shutdown will shut them down. Alas, all this too must Bless Everyone! T PS. the Buzzing the Towers was a Top Gun reference, not a Martian/Alien reference. Wait. Are you suggesting that Tom Cruise and Val Kilmer are Martians? PSS. This is part 35 of the Comet c2012 s1 ISON WTF NASA / EARTH ? series. I've learned a lot about astronomy. Now give me a big smile, baby!You share words with Me & the Astonishers on the tweet me on the twitter @NewThorThink positive them even if they are not positive action has a the Buzzing the Towers was a Top Gun reference, not a Martian/Alien reference. Wait. Are you suggesting that Tom Cruise and Val Kilmer are Martians?


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This is one of the games that i have not played so 8/10 for me as well
User Profile
United Kingdom
- Joined:
- Tue 29th Nov 2011
Recent Comments
so i better wait until next year for flip note then. so nothing for me to buy this week
for those in us want day your e-shop updates. got news for you, you will also get swapnote this week. (letterbox for us in europe helping out B-Ray 97 at the same time)
at least this one kept its promised! be out by the end of year and with two Thursdays left bingo we got news
one very happy customer (posting via 3ds due to pc issues)
want time will we get these games (i in the uk so very happy 3ds player today)?
at least that is in the open now, will NOE follow that up with news of those gba ambassador games we have been waiting ages for
I have a sneaky feeling Japan will be the 1st to know the date of the GBA free games and as it is Monday over there hopefully this week we can stop asking about Mario vs DK and Metriod fusion (two games that i have not played yet) (after all Nintendo is Japan)
I happy with the updates so far so all we need now is two things, that date for the gba games (If i was them i get the date out in the open asap to stop rumors) and letterbox


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Mary George
Mary George is an Adjunct Instructor of Music in the Belmont University School of Music. She teaches applied classical voice. She has been a vocal coach since 1978.
Mrs. George received both her Master of Music degree and Bachelor of Music degree (Studio teaching, performance and pedagogy) from George Peabody College for Teachers of Vanderbilt University. Mrs. George has completed post graduate work at University of Minnesota, Scarritt College, Northern Illinois University, Louisiana State University, and Peabody Conservatory of Music. She has participated in vocal master classes with Martin Katz, Eleanor Steber, Bidu Sayao, Thomas Grubb, Theodore Uppman, and Ned Rorem.
Mrs. George has sung in the Nashville music industry and the surrounding areas for 30 years. She has worked extensively in the recording industry as well as performing multiple times with the Nashville Symphony, twice under the direction of Maestro Kenneth Schermerhorn.
Mrs. George has received several awards and honors including Outstanding Women of America (1992), Who's Who in Education (1992), Who's Who in Entertainment (1992), two-time finalist in the Metropolitan Opera Auditions, three-time winner in the NATS Regionals. She belongs to the National Association of Teachers of Singing (NATS), and the American Association of University Professors.
Contact Information: firstname.lastname@example.com


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November 1, 2000
By Alton K. Marsh
Opportunities for viewing a space shuttle launch from your airplane have improved, thanks to an increasing frequency of flights to support the International Space Station. Flights are scheduled at the rate of one a month through the end of 2000, and will increase from an average of four or five flights a year to eight in 2001. But don't let the experience turn into a violation of restricted airspace.
The FAA invited AOPA Pilot to view the September 8 launch of the space shuttle Atlantis from a rented Beech King Air, one of two aircraft the FAA uses to intercept intruders into restricted airspace during launches and landings. The aircraft are designated Patrol 1 and 2. In addition, a NASA security helicopter is available for intercepts of aircraft that threaten to delay the launch. Our King Air was the equivalent of a police cruiser; all we needed were a red light and siren. Even the doughnuts you might find in a police car were aboard.
Just sharing the same airspace with the shuttle was exciting enough. The solid rocket boosters, used during the early portion of the flight, trailed long flames as red as those from a campfire. A plume of smoke from the solid boosters and condensation from the liquid-fueled rocket motors easily led the eye to the shuttle itself, climbing at an ever- increasing rate; it took 29 seconds for the shuttle to reach the 9,000-foot altitude of Patrol 1. Atlanta FAA spokesman Tanya Wagner, also aboard the King Air, fired her point-and-shoot camera at the unusual scene just as many of the 20 general aviation pilots then circling below must have done. Most were out of the restricted airspace, but one wasn't. More about that later.
There have been 99 launches of space shuttles—the number will reach 100 by the time you read this. Small aircraft have delayed or contributed to a delay six times. Boaters caused two additional delays. Some of the pilots stumbled into the airspace by accident, but others—the ones who now have a 30-day suspension on their pilot records—did so intentionally to "get a better photograph," FAA officials said. To the latter we say, "Get a telephoto lens!" Stay out of R-2932, R-2933, R-2934, R-2935, and warning areas W-497A and B during space operations.
If you are planning a trip to Florida (nearly all the violations are by out-of-state pilots), there are three things you need to know immediately. First, if you plan to use a world aeronautical chart to avoid the airspace visually, there is an error on the chart including Kennedy Space Center (KSC). Not all of the restricted airspace is shown, and won't be until the chart is next printed in late December. The fan-shaped portion of the airspace, correctly shown on the Jacksonville Sectional Chart, is missing from the WAC. Already, one pilot has been tricked by this and ended up explaining himself to the FAA. That same pilot received a preflight briefing from a flight service station briefer who failed to mention the launch. Be sure to ask about the restricted areas if your path takes you down the east coast of Florida. The FAA has taken steps to assure that all briefings in the future include information about space operations.
Second, while there is a lot of publicity about launches, landings are scheduled but not confirmed until an hour before they take place. This means less publicity about landings, making it even more important to check notams. While a landing may not seem exciting enough to view from miles away in a small plane, it will become so when the FAA patrol aircraft pulls alongside you in flight. The airspace is equally restricted for both launches and landings.
Finally, penalties for delaying a space shuttle launch are likely to increase. AOPA Pilot has learned that NASA officials are unhappy with 30-day suspensions handed out previously to pilots who delayed launches. They want something stiffer and are talking with FAA attorneys about their request. There are no details on what those penalty proposals might be, but consider this: A 24-hour "scrub" of a fully fueled space shuttle costs $300,000.
Here's what happens when the Air Force controllers at nearby Cape Canaveral Air Force Station identify you as a threat to an on-time shuttle launch. (By the way, launch windows for flights to the space station are only minutes long.) You will be intercepted and possibly followed to your next airport. If you are followed, that means the FAA personnel are unhappy, and it is probable that local law authorities will meet you when you land. The FAA aircraft will land right behind you.
Got the visual on that? It will be you, your airplane, police or sheriff's cars with flashing lights, and an airplane with the letters FAA in the windows—one per window—parked nearby. Not a pretty scene. Then the local newspaper reporter shows up, thanks to his police scanner. And the TV news crew, too. Got any comments for the press?
You may run, but you can't hide. The FAA followed one pilot to Spruce Creek Airport, south of Daytona Beach, and landed behind him. As the FAA pilots landed, they heard, "FAA aircraft on the ground!" on the Spruce Creek unicom frequency. "Hangar doors slammed shut one after the other," an FAA official recalled. (Perhaps the FAA personnel should have announced that they had fresh doughnuts on board.) The intruder aircraft taxied to one end of the airport and the pilot fled. The N number, of course, gave him away, and a 30-day suspension of pilot privileges followed.
If your delay of the nation's space program was just to get a better view (stupid, in other words), you deserve the fame you will receive. On the other hand, you may innocently blunder into one of the restricted zones. You may not even know when a launch is taking place. If the FAA inspector feels that the error was an innocent one, and this is your first offense, there may be no sanctions at all. Just friendly advice.
"The majority of the pilots who get in trouble are transient pilots who are just not familiar with the area," said Bob Hunt, supervisor of the Orlando Flight Standards District Office and pilot of Patrol 1. "Most of them are going to the Bahamas or South Florida." He noted that during the John Glenn mission in October 1998, two or three of the intruders "just wanted to get as close as they could get, disregarding the airspace."
Two of those who violated the restricted area—intentionally, in the FAA's opinion—to capture a good photo on their point-and-shoots delayed the countdown for 19 minutes. It took that long for the FAA aircraft to get the intruders out of the restricted zone. While most intruders nibble at the edges of the airspace and leave, one of the intruders toured the entire area from north to south. Air Force Senior Airman Amber Mitchell was watching the radar that day and recalls that the screen was full of targets. In fact, the radar's processing computer froze.
"The screen locked up on us," Mitchell recalled. "It can only process 350 aircraft, and we had surpassed that number. The vast majority of them were spectators. We had five intruder aircraft. That was a very big issue, especially in the last minutes of the count. One caused us to go into an unplanned hold. Most times they are trying to get as close as possible to get a good picture."
But during our visit on September 8, it was a different story. There were no delays of the countdown. As mentioned earlier, one aircraft was in the restricted area but was determined by launch officials to be no threat to the launch. The pilot apparently forgot that the airspace has vertical boundaries as well as lateral ones, and was at an altitude high enough to place the aircraft inside a restricted area. Patrol 1, the aircraft in which this Pilot editor rode, was 5,500 feet above the intruder. Kennedy Space Center controller Mike Long reported the aircraft to Patrol 1, but Lead Controller Ron Feile had run a threat assessment on the aircraft using special software that you can't buy from Microsoft and found that there was no problem. Patrol 1 did not intercept the aircraft. Had it done so, a picture of it would have appeared in this article.
So, congratulations to pilots who watched the September 8 launch. And yes, you were on candid Air Force radar, if not our Pilot candid camera.
Hunt asked that, in the event of an intercept, I help by mounting the letters FAA in the porthole windows. He also asked that I consider which side of the aircraft the intruder would see, and not make the mistake of spelling it AAF. (The intruder would also have seen the last letter—whatever it was—disappear periodically so that photographer Rick Fowler could take a picture.) The letters aren't easy to see. Don't make the mistake of pilots in the past who, thinking that another spectator flew the FAA aircraft, issued a friendly wave. On previous missions, the FAA taped large letters to the outside of these rental aircraft. It worked well during flight, but the tape pulled the paint off afterward.
Despite what you may have heard, neither the FAA nor NASA has any interest in shooting an aircraft down, one official said. The rumors about "resources" available to NASA, the Air Force, or the FAA that involve use of force against civilian aircraft are untrue. (Yes, the NASA security helicopter has armed security forces aboard.) Our patrol was more like fish spotting than combat.
Don't get the impression that NASA wants nothing to do with general aviation pilots. When there is no space launch in progress, you'll find that it may be possible to fly over the space shuttle runway at 500 feet. Assuming that KSC Tower controllers do not have a caravan of heavy transport aircraft inbound, they just might approve such a request. Call them first on 128.55 MHz when the KSC tower is operating (from 8 a.m. to sunset). After that, Daytona Approach handles the KSC area and won't let you get as close to the shuttle runway. Careful, though. Immediately east of the shuttle runway lies continuously restricted airspace.
Why is it so important that officialdom interfere with your fun? The space shuttle Atlantis needed to reach the International Space Station, a fast-moving target, and had a launch window that was only a few minutes long. Mission commander Terry Wilcutt and his crew of six, including two Russian cosmonauts, trained hard for their mission. They obviously would prefer that nobody delay it. An incident, no matter how brief, would eat up the entire launch window and cause a 24-hour postponement. But let's not blame it all on privately owned aircraft. Boats are the bigger concern; their skippers may be willing to vacate the restricted area in the Atlantic Ocean when warned, but can do so only at slow speeds. That's why military support personnel stayed up all night on September 7 to clear the seas.
In addition to tougher sanctions against pilots violating the airspace, NASA and the Navy are completing a plan to deal with terrorist attacks on the space shuttle. The new plan should be ready in December. No details are known and probably will never be announced. But it is a good guess that pilots should be especially careful not to trigger the military's antiterrorism capabilities.
Some of the reasons for avoiding the airspace should be obvious. The first is that you don't want to be hit by the shuttle, becoming the first Cessna 152—or parts thereof—to achieve orbit. It's fast, and your see-and-avoid tactics won't work should the shuttle take a wrong turn. But less obvious are the needs of support aircraft.
Nearly 100 personnel, 50 of them military pilots, attended a space shuttle support briefing at Patrick Air Force Base near Melbourne, Florida, a day before the launch. Most of them operate military rescue aircraft that will take the most direct route possible to the shuttle should there be an accident. They need clear airspace in which to work. Two thousand KSC employees and 650 military employees had gotten up in the middle of the night to support the launch. You don't want to be the civilian pilot who makes them do that two days in a row. They'll wake up taking your name in vain.
A NASA Gulfstream II aircraft, designed so that it is able to land with a steep angle of descent like the space shuttle (see " No Go-Around," April 1999 Pilot), made repeated circuits of the area and several landings on KSC's space shuttle runway just minutes before launch. The purpose was to assure that the cloudy weather would not be a problem. Should the shuttle have an emergency, it might return to KSC. Frankly, it was a tight fit just to have Patrol 1 and the Gulfstream II in the restricted area.
Do we have enough reasons listed yet for not testing the patience of three government agencies? Here's a long shot, but one worth noting. That plume left by the solid rocket boosters is toxic and filled with aluminum particles. After the September 8 launch it drifted over KSC; space center workers were told they had one hour to wash their cars to avoid damage to the chrome. One worker didn't make it and has tiny pits in the chrome. Once the cloud disperses, it is harmless. But you wouldn't want to fly through it; it would cause a slight burning sensation on your skin.
While all the reasons above are more than enough to make you want to stay clear, they can't prevent inadvertent intrusion.
"It's complicated airspace," Hunt said. "We recommend that pilots stay four miles west of the [Indian] river that borders the airspace, and they will stay out of trouble."
Another suggestion is to read the instructions on the Jacksonville Sectional Chart. You'll find helpful frequencies, phone numbers, times when the areas are most often activated, and methods for looking up notams concerning launches. Notams are listed under Melbourne (MLB). The chart also explains that the darker tinted areas off Cape Canaveral and over Merritt Island are FAR 91.143 areas. That regulation says: "No person may operate any aircraft of U.S. registry, or pilot any aircraft under the authority of an airman certificate issued by the Federal Aviation Administration within areas designated in a Notice to Airmen (notam) for space flight operations except when authorized by ATC, or operated under the control of the Department of Defense Manager for Space Transportation System Contingency Support Operations."
For the latest information on future space shuttle flights, try an Air Force Web site ( www.patrick.af.mil) or, for general information, see a NASA Web site ( http://spaceflight.nasa.gov/index-m.html).
Only Patrol 1 and Patrol 2 are afforded the best seats in the air for space shuttle launches. No general aviation aircraft will get closer. There are two ways to be that close to the space shuttle: Get to know Bob Hunt and ride with him, or violate the restricted area and then get to know Bob Hunt.
Links to information about space shuttle flights can be found on AOPA Online ( www.aopa.org/pilot/links/links0011.shtml). E-mail the author at firstname.lastname@example.com.
AOPA Pilot Senior Editor Alton Marsh has been a pilot since 1970 and has an airline transport pilot certificate and instrument and multiengine flight instructor certificates, aerobatic training, and a commercial seaplane certificate.
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GARDEN VALLEY — Golden Sierra hosted and competed Wednesday alongside six other schools in a successful afternoon at the second Sierra Delta League track and field meet, Grizzly team took first: the girls edging Highlands 130-126 and the boys topping Vacaville Christian 139-102.
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GARDEN VALLEY — Golden Sierra hosted and competed Wednesday alongside six other schools in a successful afternoon at the second Sierra Delta League track and field meet, Grizzly team took first: the girls edging Highlands 130-126 and the boys topping Vacaville Christian 139-102.
Becca Brown, continuing her senior-year dominance, placed first in the 400 meter (68.86) and added another victory and in the 200 meter (28.10).
The Lady Grizzlies produced 20 points by capturing firsts in the 4x100m and 4x400m relays.
Amanda Troiana led off the 400-meter relay, followed by Katie Robertson and Mia Belloumini. Brown anchored and helped her team to a winning time (54.21).
Brown sealed a solid afternoon of sprinting in anchoring the 4x400m relay, along with Alexis Babbes, Troiana and Maylene Coetzee that combined for the top time (4:56.78).
Babbes closed out the winners by clearing 8 feet in the pole vault.
The varsity boys' 3200m marked a strong showing with a Ryan Sewell win of 10:51 while teammate David Blanton came in second in 11:18. Sewell clocked 4:56 in the 1600m for the day of double wins and Blanton crossed the line in second (5:05).
Alex Soden stepped on the track for the first time this season dominating the 110m high hurdles in a time of 16.95. He also finished third in the 300m hurdles in 53.3.
Patrick Lewis picked up a first in the pole vault with a height of 9-6 while Brandon Susic secured second place, clearing 9-0.
Hunter Hughes was second in the 400m and 800, clocking 58.40 and 2:17.58. Dominic Chamberlain leaped 17-06 in the long jump for a personal record and good enough for third place.
The Grizzly boys got help in the team scoring with a third in the 400m relay event with Soden, Lewis, Hughes and Chamberlain passing the baton around (48.18).
Carlos Gonzales found his groove in the shot put with a toss of 41-08 for second place. He then unfurled a huge throw of 102-08 for third place to extend his new PR by over 6 feet.
The Grizzlies will continue competition this weekend at the Bret Harte Invitational.