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PcM, PjM, CE Passed!
Woo! I just got the official pass report and wanted to share my experience, since reading the experience of others was a major factor in me getting through these.
First the background: I have 3.5 years experience in IDP/AXP, starting back in 2013, with a 3 year break starting in 2015 for grad school in Architecture since my undergrad was in Music. I graduated this past May and started the test process in July.
Most of the time spent at first was looking on this community at others and their processes, which helped me get a good frame of reference for my own goals and schedule, as well as what materials I would need to acquire to study. I started studying in earnest around September 15th after deciding to start with Practice Management, then go to Project Management, followed by Construction and Evaluation. So it ended up being PcM on October 13th, PjM November 16th and CE November 21st. Didn't need much time between PjM and CE after all the overlapping studying I did.
ARE 5.0 Handbook - Seriously, they actually tell us what they're testing us on. Use this as a your first and last resource, read the section objectives, and make sure you understand the content. This is what the test is on, they aren't trying to trick us.
Black Spectacles - I watched the videos generally over breakfast and lunch, mostly at 2x speed playback but slower if I wasn't sure of the content. The best part about BS for me was the practice exams. They are the same format as the actual test, and I found them to have a few questions that were relatively challenging. The practice exams would force me to think through a question, then if I got it wrong, find out why. The challenging of my knowledge and re-shaping it was the best way for me to learn. If I got an answer wrong that I really thought about, it made the correct answer stick all the more. They also have a podcast/video series that is helpful for the mock exams as a refresher, and for the miscellaneous ones just as a confidence booster.
Architect's Handbook of Professional Practice - Totally invaluable resource for PcM and PjM, using Kevin Griendling's guide followed by the Narmour Wright guides to refresh. I dove deep on the book mostly for PcM, and passed the book to a colleague, so I used an old office copy of the 13th edition for the other exams. There isn't a guide for it, I just looked at the table of contents and used my practice exams to be the judge for where my knowledge was lacking.
Mike Hanahan's Lectures - All of them. They're all great for PcM, but I only listened to the contract ones as I moved to PjM and CE. I basically listened to them on repeat until I knew every anecdote he was about to tell and why it related to the contract.
Tatyana Aksamentova Study Guides - Much thanks to her for putting together really concise and smart guides for these tests. I used them in the last few days of studying for each exam to refresh my info and make sure I had my bases covered. I'd say most of what is on those study guides were on the tests. Don't use the guide as the sole resource, just as a good benchmark.
Brightwood - I used this for PjM, and only skimmed 3 chapters for CE, since there is so much overlap. It wasn't a great resource for me, but I wouldn't not recommend it, only because I think getting info from as many sources as possible is very valuable.
Colleagues - I can't stress this one enough. If I had a question about something in my studying, asking another test taker at the office was great because we could both work through the problem. I would also ask at least one question per day from a licensed architect or admin or someone in the office that had to do with my test. Getting real world situations and stories is invaluable to planting the information more firmly in my brain. I'm also at the same firm where I worked before grad school, which makes the whole thing smoother - everyone is supportive and rooting for each other.
My Wife (and cats) - She is a queen, a beauty, a joy, and the best companion to have in this time. She gave me the support and space I needed to put my head down and work, and also to come back to and unwind. The cats helped by distracting me when my head was in the books for too long, and being the lap warmer I needed for those long hours at the desk.
I put in about 45 minutes every morning over breakfast, 30 at lunch, and at least 1.5 hours every week night, with about 10 hours each weekend. I ended up doing a practice test every week, at least one, often more - primarily the BS exams, but I also threw in the Designer Hacks free tests as another benchmark. The best score I got on the BS exams was in the 75-80% range, which sounds low, but I guess it worked out anyway. I'm also weird in that I don't like to cram, so I take the day before my exam to do anything except study, and I prefer to take them before the weekend for the same reason. I don't think cramming helps me at all.
The Test:
Make no mistake, these tests are very hard. The big difference between the practice and the real thing is the way the questions are structured. The BS exams were tricky, but the real questions have more reason and thought required. For CE particularly, there absolutely were sections and diagrams that surprised me, but I took a deep breath and realized everything I needed to know to answer the question was right in front of me.
My preferred method is to check the case studies first to look at the resources, then run through the multiple choice. Someone had mentioned that the highlighter and strikethrough functions weren't working, but it worked perfectly for me on all except the case study scenario box. I highlight and strike liberally, together with marking, so that when I come back to the question, I already have a few answers marked out that I knew were wrong.
I make sure I get to the case studies with 1.5 hours to go, but generally get there sooner, and get through all the questions with about 45 minutes left. I spend almost every minute left reviewing and reviewing. I read the questions with the mouse cursor, click-dragging along so I make sure I read every word and highlight the important ones. Each word makes the sentence.
That's really about it. I hope this could be even remotely as helpful to others as this community has been for me. Next up is PA, then PDD and PPD!
12 comments
Besides the above listed resources you listed above, did you use Kaplan/Ballast as Tatyana states for her study guide? In addition, for the BS I'm assuming you did the monthly subscription for the Pro. I'm going to start studying PcM starting Monday and use your suggested methods. Did you study PcM, PjM and CE one at a time?
I used Brightwood (which I think is part of Kaplan if I understood it correctly) for PjM, and skimmed it for CE. I didn't think it was a particularly helpful source, especially if it were my only resource. As an extra, it helped marginally to reinforce concepts I had gotten elsewhere.
My Black Spectacles subscription is "Premium," I think, so I have access to videos and practice tests, but not flash cards or their "Practical Applications" videos. I had access to those for a trial and did not think they were very helpful. There is a free app on Android and probably iPhone called Quizlet that I use. Apparently loads of ARE takers use it too, because they have shared their flashcards, so I referenced those every once in a while. I didn't use flash cards much though.
I didn't study for all three at the same time specifically, but the resources overlap. The Architect's Handbook of Professional Practice has content for all three exams, so if you're reading it, you're learning things that will be pertinent to other exams. I should emphasize that they are not testing the same things, the subjects are quite different.
That said, with the overlap, once you read the AHPP using some of the guides for PcM, you can then focus your review of the book for PjM or CE. The same can be said for the Hanahan lectures, listen to them all for PjM, but for PjM and CE, hone in on the subjects that are more pertinent to those exams. I did end up taking CE 5 days after PjM, but I don't necessarily recommend that.
Good luck studying!
I took two (2) practice BS exams and did horrible but what it did show was that I approved in previous sections. I still have this upcoming week then my exam is on Monday. The BS questions are quite challenging, but it also shows me where I need to improve. I plan to take probably the remaining two before Monday. I tend to go back and review the sections from the questions that I got wrong and continue to read and understand the concepts. Any thoughts?
Don't let the stress get to you. I know the scores can be intimidating, but stress can do really rough things to you as well. Try to focus on the areas you know you need work, and don't let a number from Black Spectacles worry you. I would also use what they say you need work on as a guide more than a strict law to follow; each time I took a practice test, I did worse in some areas and better in some areas, but I knew where I was weak from a gut feeling.
I personally found the BS practice tests quick enough to take on a weeknight, so if you have time, I'd recommend taking at least one more, but your strategy might be different too.
Best of luck!
I took PcM today for my first test and I got a Preliminary Fail. I found myself stumbling on a lot of questions, thinking too myself did I even read that. I used the ARE Handbook as a guide of what to study and I studied those items. I put in about 20+ hours a week since Jan. Today, I take a break and tomorrow I'm back at it while the information is still fresh. I'm thinking about studying for PjM and take in Mid-April (about 1 month) then CE. Thoughts, on which one I should take next? Any suggestions will help.
Hey Michael, sorry to hear about PcM, but don't be discouraged. I don't think it is unreasonable to forge ahead to PjM, as you have likely already at least glanced at most of the material for that test. I'd highly recommend looking up the Schiff-Hardin lectures if you haven't and dive deep into the contacts. That should set you up nicely for PjM with the AHPP, which all leads into CE. Don't forget they are very different tests though. Keep referring to the ARE handbook to see the differences in how they refer to the different tests. Best of luck!
Yes, I do have the Schiff-Hardin lectures and I will print out the contracts that are listed for PjM on the ARE handbook. I did notice at the other posts were discussing conversions. Do you have an outline for PjM or remember another post that had a good outline for PjM. I'm going to print out Tatyana study guide for this section. I start back studying today.
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Red Hot Boilerplate: How to Make the Start of a Course Interesting
Most online courses start with a section of boilerplate that outlines the structure of the program, the rules of the course, and provides other information that usually doesn't vary much from course to course within a curriculum. With any luck, students will peruse the pages when they begin a program, but it's unlikely that they will spend much time on them, let alone revisit them for subsequent courses, especially if they are exactly the same as those they've previously encountered.
Boilerplate is a little like the safety announcements at the start of a flight: vital information that we all ignore, even at our own potential peril, because we think we've heard it a thousand times. If you've taken a flight recently, however, you may have noticed that some airlines are trying to capture the attention of their passengers by making entertaining safety videos that people will actually want to watch.
Instructional designers would do well to take inspiration from the airlines' tactic and look for ways to get students to pay attention to important material, even if they've encountered it before. Since the start of a course sets the tone for it, designers can impart a sense of levity and ludic interaction before an earnest educational experience. In another blog entry, I talk about the usefulness of humor to keep students engaged, a theme that also threads itself throughout the suggestions below. Although online courses should not need to distract and placate students the way airlines do their customers, the introduction of a course presents an often overlooked opportunity to get students excited about the learning to follow.
In addition to acting as preparation for the substance of the course, introductory materials also provide an opportunity to cultivate your presence as an instructor. Even if the content is standardized, how you introduce the material and what you ask students to do with it can give them some sense of who you are as a person, as the examples that follow demonstrate.
Elements of an Introduction
The first step to making the course introduction interesting is to decide what information you need to convey and how to organize it. Potential subjects to address include:
- An introductory welcome to the degree program and the department in charge of it.
- An overview of the degree program's requirements, including an explanation of how the particular course fits into the curriculum.
- A description of how the course works, i.e., an explanation of the way it is organized, recurring due dates, learning outcomes.
- Expected time commitment per assignment, per week, or for the course as a whole.
- Expectations for behavior, particularly standards for communication, collaboration, and community formation.
- Information about institutional resources available to the students, such as libraries, software, or databases the program has purchased, etc.
- Directions for submitting work. This may include required file-naming conventions, instructions on how to use tools like Google Docs, etc.
- Welcome discussions, where students introduce themselves to each other.
- Ongoing fora where students can confer and seek help.
The above information should be in addition to that found in the syllabus, which itself has a different function (and is a topic for another post). Although there may be some overlap between the introduction to a course and its syllabus, preliminary materials should be presented in a less dense and more interactive way with the goal of familiarizing students with the course, whereas a syllabus usually sets parameters and acts more like a contract between the instructor and the student. Note that the above list is meant to be suggestive; academic programs most likely already have introductory information at their disposition.
Getting Students to Think About the Course
Once you've established a list of what needs to be accomplished in the introduction to a course, the next step is to follow the tenets of good pedagogy and consider the learning goals attached to each task. If, for example, you want students to plan for fitting classwork into their existing schedules, you can design an activity asking them to do so. Requiring students to submit their schedules is neither the instructor's right nor necessary, since public knowledge of the class's extracurricular time commitments is not the learning objective. The stated goal of getting them to plan (and verifying that they have done so) can be accomplished by requesting such information in a discussion topic, for example. The following prompt requires students to examine their schedules without asking for too many personal details:
As you have seen in the description of the work for this course, it requires approximately five hours of reading and five hours of writing or other work per week. This is an admittedly significant commitment that requires thinking ahead to accomplish. One of the goals for this course is to form a community of support that will help its members learn. To that end, please write a post addressing the following:
1. What advice would you offer to your colleagues that you have learned from previous educational experiences, online or not?
2. What will be the biggest challenges to getting your work done? How do you plan to overcome them?
3. Describe an assignment from a past course that you found hard to complete, perhaps because of time limitations, and explain how you did so.
Although the above discussion post addresses issues of overcommitment and scheduling directly, responding to it also asks students to think about how they will meet the rigors of the semester more generally. Mutually sharing this information will allow students to get to know each other as individuals, not just as classmates.
(Note: For a more efficient, but less reflective and interactive means of getting a sense of students' availability, free websites for scheduling, like Doodle, or for surveying, like Survey Monkey, can be useful for synchronous meetings or live lectures.)
Getting Students to Remember the Details
If the adage that the best way to learn something is to teach it, then asking students to act on what they learned must be the second best. Impending assessment is a typical, but nevertheless effective way to get students to study material. (Moreover, quizzing students in the introduction to a course will introduce them to the assessment tools before they are responsible for learning any course material.) Although it could seem condescending to test students on information about the course, completing a cleverly designed activity about such material can be fun. Quiz builders in Canvas and Blackboard allow instructors to pose many types of questions and to import a variety of media, which allows for great creativity. A humorous way to teach students netiquette would be a video showing how not to interact with class members online, and then asking them to take a quiz on the subject. They could, for example, comment upon a video dramatizing how not to act during a live web-conference, or analyze humorous, but poorly written emails, as in the video "Email in Real Life" below. (Please note that subsequent videos are not relevant to this article, although YouTube may start playing them automatically.)
Show and Tell Me
Some introductory material may be too detailed or too abstract to ask students to learn before having to do so for the course. Accessing and identifying peer-reviewed articles through the institution's library site, for example, requires not only many steps, but an understanding of what you are looking for. Modeling multi-step or intricate tasks through screencasts or video tutorials and then asking students to repeat the task themselves has the dual benefit of providing a resource to which they can return, as well as accommodating various learning styles. It also is another chance for instructors to make their presence felt, since even something as simple as the professor's voice individualizes the course, as shown in the video below on how to find peer-reviewed articles at the library made by Professor Susan Krauss of Concordia College New York.
As pointed out, asking students to do something with what they have learned is an effective way to get them to retain it. Revealing the value of what they are doing will make introductory assignments seem even more worthwhile. Susan Krauss's video on navigating the resources of the Scheele Memorial Library could, for example, quite naturally lead to an assignment on how to do research and compile a bibliography related directly to the subject of the course. Generally speaking, it is always good strategy to make the purpose of assigned work readily apparent, which means tying as many elements of the introductory material to the actual topic of the course as possible.
Tying It All Together
If there is a common thread to the above suggestions, it is the idea that meta-information is no different from subject-matter material in terms of pedagogy. In other words, use good teaching principles when presenting even standardized content. Boilerplate's parallel status to the primary course material leaves room for fun and experimentation to capture students' attention. Although serious in intent, introductory material can still be presented in a manner that will make students will find it engaging and worthwhile.

subset: commoncrawl


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According to the Federal Trade Commision, debt does not go away when the debtor dies (Debts and Deceased, 2011). When a person passes away all of their assets and liabilities go into the person's "estate." Essentially, the estate is liquidated and any money within the estate is to pay-off the person's debt. If there is not even money in the estate to pay off the debts, the debts will go unpaid; unless there is a living co-signer or spouse (Debts and Deceased, 2011).
The Nutrition Labeling and Education Act of 1990 (NLEA) established labeling regulation on food packaging to require the Nutrition Facts label (Martinez, 2013). In an effort to lure consumers to purchase its products, companies will add voluntary health and nutrition related (HNR) claims on their products. These claims include "low fat," "no sodium," etc., (Martinez, 2013). This type of labelling is in hopes that consumers will buy the product. The U.S. Food and Drug Administration (FDA), is in charge of determining what HNR's can be used.
- Estate: all the money and property owned by a particular person, especially at death (Oxford Dictionaries).
- Debt: a sum of money that is owed or due (Oxford Dictionaries).
- U.S. Food and Drug Administration (FDA): an administration responsible for protecting the public health by assuring the safety, efficacy, and security of human and veterinary drugs, biological products, medical devices, our nation's food supply, cosmetics, and products the emit radiation (Usa.gov, 2018).
Debts and Deceased Relatives. (2011). Federal Trade Commission Consumer Information. https://www.consumer.ftc.gov/articles/0081-debts-and-deceased-relatives
Martinez, S. ( 2013). Obesity and Other Health Concerns Lead Food Companies to Step Up Health and Nutrient Claims. USDA. https://consuminghealthmatters.files.wordpress.com/2016/05/healthconcernsleadtolabelclaims.pdf
Oxford Dictionaries. (2018). https://en.oxforddictionaries.com/
Usa.gov. (2018). Food and Drug Administration. https://www.usa.gov/federal-agencies/food-and-drug-administration


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Sometimes you turn up for work and you know, it is a good day to be doing what you love to do.
As soon as I drove up the driveway and saw the beautiful Homestead again, I knew it would be a little bit of me! I had looked at Mataia for my own Wedding and was already working out in my brain the rooms with the best light, the spots I would make work for them.
It is always a little interesting, photographing a couple you have never met before – but Leila and Charl were warm and welcoming, their families and friends were buzzing around putting the last little touches on things when we arrived. We got straight into it with the boys who had us laughing from the get-go, it's always fun when the team are in good spirits.
As I headed down the stairs to the girls – I could hear someone humming a tune and busted Uncle having a sneaky dance near the kitchen sink hehe
Leila was a confident and beautiful bride – her amazing style was stamped on everything to do with their Wedding, you could tell from the moment I walked in she was SO READY to get into her dress!
So we took it downstairs where there was more room and beautiful light
Just as Leila was about to walk out I realised she hadn't put her earrings on – she was super happy I remembered as you can see.. she popped them on just before walking out to say her vows and see their guests, it was all on!
I will NEVER get sick of petal confetti EVER EVER EVER Three generations of beautiful women x
WeWe headed back to the venue and it was ALL ON – I am partial to a bit of dancefloor action, and had to laugh at the speeches (the bits I could understand anyway).
I had so much fun..so many laughs and felt the energy of all of those around you who were SO excited for the next chapter of your lives together.
Thanks for having me get right in there to document the real deal feels alongside Jodie for your special day at Mataia Homestead!
x Bianca


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British design for Kids and Grown-ups
Corby Tindersticks is an illustration led lifestyle brand based in Greater Manchester. Our products are designed to be fun and creative, offering a point of difference to the high street. They are inspired by vintage teapots, Enid Blyton books and Northern soap operas.
All our products are made in the UK. Our fabrics and print inks are locally sourced and eco friendly where possible. We also print on recycled Paper where possible.
Corby Tindersticks was founded in 2012 and has developed into a successful brand being stocked in stores and galleries around the world.
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Please see notonthehighstreet.com's delivery policy.
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18 Ainsdale Grove, Stockport, SK5 6UF, GB
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Follow Us:
Glee was under the spotlight on the first day of the show on BBC's Breakfast programme. Fronted by presenter Steph McGovern, leading garden industry figures were interviewed during a live broadcast from the halls.
Regular updates from Steph featured throughout the three-hour show, with talking points that included the latest gardening trends and insights, new product launches and debate on how to engage with the younger gardener.
Duncan McLean, buyer at Wyevale Garden Centres, was interviewed by Steph on the garden retail market, whilst Sheila Hill, UK general manager of Scotts Miracle-Gro also did a piece to camera discussing the latest trends.
Romeo Summers gave viewers a tour of the new Retail Lab @ Glee and its interactive hubs focusing on Family, Community, Re-Wilding and Well-being and how the public can be inspired by gardening with the latest products. George Williams of SmartPlant also gave a demonstration of the SmartPlant app, innovative technology that enables the consumer to identify plants and receive advice and how to care for them.
The coverage also included Matthew Appleby of Horticulture Week, who was interviewed from the industry viewpoint.Trevor Pfeiffer, director of GTN, was interviewed twice about Glee New Products. He was spotted on TV by son Robert and grandson Arthur (aged nine months) - see picture below.
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text: I have installed UTAU multiple times by now, wiping everything clean. No matter what I do, it just won't work. The interface is in English, although everything else is just gibberish. Here is what I have tried, with restarting between steps and wiping clean folders of any traces before and after reinstalls: - Set Locale to Japanese and installed Japanese language pack. - Reinstalled UTAU 0.4.18e from zip. (about 3 times because I thought I messed up) - Unblocking installation before extracting the file - Turning off Windows Defender while downloading and installing. - Run UTAU as Admin - Giving Full Control to all Users (since this is my own personal PC) - Using tutorials on Youtube, this forum and other places on the net. - Trying to troubleshoot it myself. - Running it in Windows 8 and Windows 7 Compatibility mode. - Shutting down computer and starting it up again in place of restarting. - Installing UTAU somewhere else instead of the program files folder. What happens is that the program looks fine, although when I try to put a note with the pen, it turns into a question mark. When I try to manually put in a note like say, "が" (ga), it turns into "?", sometimes "??". I can add romaji fine, but nonetheless, a note won't play. I have a Windows 10 64-Bit OS with Windows 10 Home Edition, 1803 Version. Unless Windows messed with the permissions, I haven't changed anything unless stated above. Any kind of recommendation is welcome, I never had this issue before I started using Windows 10, on a clean install nonetheless. I will provide screenshots if necessary.


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This weekend I took my day off. I slept in, jogged the dogs, watched the first day of Premier League football, did some garden tending and food preservation, and I cleaned out the freezer.
Not the fridge freezer – the big chest freezer. The bottomless pit. The point of no return.
Over the past few weeks I've been working my way through the food that's been in there, and have been sharing some with friends, and with only a few packages left, I cleaned out the beast. Defrosted it, washed its walls, everything. It was almost like new again!
So – that's been done, now I'm ready to start filling it again. Which is good – I was blessed with significant food gifts at church today, and my garden is growing abundantly, and harvest is almost here. So I'll need somewhere to put it all!
As I was cleaning the freezer, I was thinking of the whole process as a metaphor. When we're putting away our food in the freezer, we're putting good stuff in there – we wouldn't intentionally keep something we don't like or didn't intend to use. Yet with time, more and more food piles up and some of it falls to the bottom and it gets forgotten, or eventually wasted. There is a limit to how much we can put in the freezer – both in terms of space and in terms of our capacity to use what's in there. Some times there's so much stuff in there that we can't access what we might want, or we can't even remember if it's still hiding in the depths. Our freezers also show us how we categorise things; sometimes starting out organized but falling into chaos and disarray, sometimes staying neatly compartmentalized.
So how we use our freezers can show us a bit about how we live our lives…
We often store a lot of good stuff, memories and stories and intentions. It's all good stuff, but unless we're using it and applying lessons learned to our lives, it's just sitting there taking up space. We're meant to grow and develop and learn, but we need to use that good stuff that we've got. If our spirituality becomes jammed into our live with everything else, it can fall to the bottom (as it were) and be forgotten – which will not do us any good.
Our lives are also restrictive in just how much we can do, and so we are challenged to make our priorities. If we jam our lives full with so many activities where we do not focus on our spirituality, then when the time comes for us to try and tap into our spiritual selves, we won't have much to find. We're challenged to always have some focus on the spiritual component of our lives, and to allow God space and time to enter into our lives.
Our spiritual development may have been extensive in the past, but has been in disuse – hiding in somewhere with the rest of our lives – that we don't know where to find it, or how much is there, or even how to use it if/when we do find it: just like those forgotten treats in the freezer.
And our lives are sometimes very organized, at other times they are chaotic. If we require organization, are we being flexible enough for unforeseen experiences in which the Spirit may move us? If we are in constant disarray, would we even know if the Spirit was calling us through the melee of everything else? Is our emphasis on Spiritual nourishment a high enough priority for us to keep it separately organized, like sitting in the top of a freezer basket?
And, most of all, are we using our Spiritual freezer for it's best purpose –as a tool to preserve something nourishing and positive until we're ready to use it, but aware that it must be used if we are going to create space for the next new development.
I don't know if I have all the answers for myself, but I pray that my capacity for spiritual growth will be as prepared as my freezer is during this season of abundance.


text:
Registered 2011-06-21 by Amit Kucheria
Powerdebug allows developers on embedded platforms to view their clock tree hierarchy, regulator status, etc.
Project information
- Part of:
- Linaro PMWG Project Group
- Maintainer:
- Linaro Power Management Working Group
- Driver:
- Daniel Lezcano
- Licence:
- Eclipse Public Licence
View full history Series and milestones
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trunk series is the current focus of development.
All code Code
- Version control system:
- Bazaar
- Programming languages:


text:
Unity Trust Bank attracts key sponsor for Credit Union Awards
Unity Trust Bank is delighted to announce MasterCard Worldwide as the major sponsor of the 2011 National Credit Union Awards for Excellence.
Now in its second year, these unique awards are open to any credit union in the UK. They were introduced to champion and pay tribute to the extraordinary work of the countless unsung individuals who give their time and expertise to the credit union movement, many on a pro-bono basis.
In addition to supporting the awards, MasterCard Worldwide is sponsoring a new, individual category award, for Innovation. This category was included to offer credit unions an opportunity to showcase how they have introduced inventive and original approaches to benefit their membership.
Matthew Lanford, MasterCard Worldwide's Head of Prepaid Europe, said, "We recognise Unity's history of supporting credit unions and see these awards as a natural fit with our own commitment to encouraging financial inclusion. As a corporate citizen, we are constantly working to improve the communities in which we live and work and we see our involvement as a very positive way to contribute to society throughout the UK."
John Brooks, Unity's Sales and Marketing Director, said, "This sponsorship marks a very important moment for the Credit Union Awards. We're delighted that MasterCard Worldwide has come on board to support our initiative which, at its heart, is a celebration of the invaluable contribution delivered by credit unions across the UK in tackling financial exclusion. We are looking forward to working in partnership with them to make the awards even better than last year!"
Last year saw an impressive calibre of entries submitted by a broad cross-section of the credit union movement.
Entries are now open for the 2011 National Credit Union Awards for Excellence. For more information and to see the categories visit www.creditunionawards.co.uk.
Notes to Editors:
Unity Trust Bank Contact: Adrian Oldman, Head of Marketing and Communications, 0121 616 4149 / 0771 424 1648.
About Unity Trust Bank
Unity Trust Bank ("Unity") is the leading provider of banking services to civil society organisations, social enterprises, trades unions and small to medium sized enterprises.
Unity was established in 1984. The organisation has a deserved reputation for its knowledge of the particular requirements of its customers and the delivery of a first class portfolio of services.
Unity has an impressive track record of working in partnership with many organisations that campaign for progressive social change and develop new and innovative ways of improving civil society. It also works in partnership with statutory bodies.
For more than 25 years Unity has worked with its credit union customers and trade bodies to develop initiatives to help make a positive and lasting difference.
Unity's commitment to credit unions is evidenced by the fact that almost 45% of all credit unions choose to bank with them, and a number of Unity staff are both members of and on the board of credit unions in their own right. In 1997, Unity joined with one of its trade union shareholders, the Bakers Food and Allied Workers Union, to help them form a credit union of their own. This successful credit union was the first trade union credit union in the UK to have a national common bond. Since then, Unity has advised a number of other trades unions such as the GMB, PCS and RMT on how to establish credit unions of their own. And, since it was launched in 1998, Unity's Credit Union Development Fund has put more than £100,000 to the movement through grants to individual credit unions.
About the Awards
The National Credit Union Awards for Excellence from Unity Trust Bank were devised to recognise outstanding contributions to credit unions.
Credit unions are co-operative financial institutions, owned and controlled by their members and operated for the purpose of promoting thrift, providing credit at reasonable rates, and providing other financial services to their members.
Credit unions differ from banks and other financial institutions in that the members who have accounts in the credit union own the credit union, electing their board of directors on the basis of one-person-one-vote, irrespective of the amount of money invested in the credit union.
Since July 2002, UK credit unions are regulated by the Financial Services Authority.
The categories for the 2011 National Credit Union Awards for Excellence are:
• Active partnerships
• Community mobilisation
• Credit union education
• Credit union innovation
• Fundraising - raising additional capital
• Fundraising - on behalf of a charity
• Marketing
• Photography
• Single benefactor
• Young leader of the year (17-35)
• Young saver of the year (under 16)
Filene Award
The judges will choose an overall winner from one of these categories to be the recipient of the Filene Award for Social Entrepreneurship, named in honour of Edward Filene, one of the 'founders and benefactors' of the credit union movement.
This award will be presented to the person or credit union who has done the most to promote the concept of credit unions.
For further information about Unity Trust Bank please visit www.unity.co.uk.
This press release was distributed by ResponseSource Press Release Wire on behalf of Unity Trust Bank in the following categories: Business & Finance, Public Sector, Third Sector & Legal, for more information visit https://pressreleasewire.responsesource.com/about.


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We finished the pet shop. Here are some pictures of the finished building. this one has 2 buildings. a pet shop and a apartment building.
here is a shot of the 1st floor of the apartment
This was the second story of the apartment. we had a little guy in there painting the walls 🙂
the front of the apartment
balcony on the top floor, rear of the apartment building
ground floor of the apartment building. this building had a cellar door
this is the second floor of the pet shop. here we have a coffee pot, sink, stove, fireplace and a spot to eat
here are shots of the 2 buildings side by side.
Coming up: the Emporium


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According to the scriptures, to live in Christ is the opposite of living "in the world." Hmmm. Anyone confused by that? Since we are living on planet Earth, don't we have to live in the world even if we are Christians? Aren't we going to have to compromise somewhere, sometime? We know the world is full of wickedness. We know we are going to run into evil pretty much every day – or it will run into us.
Did Jesus live in the world without compromising? Yes, he did. He came as and remained as the sinless, spotless Lamb of God throughout his life, and on the cross. How else would he be able to bear the sin of the whole world when the Father placed it upon him, to be our sacrifice for our sin, for all time? He had to be perfect. The tiniest compromise would have made him imperfect, and unavailable for God's plan of salvation for mankind. Only God himself could complete that plan, so he sent his Son.
When I listen to God's plan to save us with my spirit, I get it, praise the Lord. If I listen with my mind only, it's just a big boggle. Knowng this makes it clear why speaking the gospel to unbelievers must be baptized in prayer, asking God to reveal the truth to them. Well, that's off the main point of this blog post; consider it a free point!
To live in Christ requires Christ living in us. Yes? You agree? When we are saved, the Holy Spirit, who is the Spirit of Christ, literally moves in and takes up residence in our hearts. One way we recognize his presence is that now we can understand the Bible and it is definitely living and active in us. The Holy Spirit teaches us and comforts us and speaks to us whatever he hears the Father saying. We can ask for information and listen to the Spirit of God for an answer. He lives in us so we can live in Christ.
What about compromises; that is, living with one foot in the world? Suggestion: read the epistle of First John, toward the end of the New Testament. There is great wisdom in that book about not compromising, and the consequences if we do. It explains how to know which people are truly in Christ, and who is not in Christ.That is good help for forming relationships among people, because if you hang out with the wrong people, you will certainly compromise with evil to maintain those loyalties. Those people can be your own family members, and then you need to get more of Jesus living through you to handle being with them but not compromise with them. Some people go to jobs every day where there is enormous temptation to compromise Christ to fit in. It can get scary.
Christ is perfect. If we knowingly compromise on any little thing, we are shutting him out. If we sin unknowingly, he will lead us to repentance, because he does not want us to be alienated from himself in any way. God will provide new strength every time a temptation arises to act like antichrist to fit in to this world. We have the ability to discipline thoughts so that every one is filtered through the eyes of God. This discipline comes about by learning God's Word, the Bible, and prayer. If you can make the effort to turn on the television, you can make the effort to turn it off and open your Bible instead.
We abide in Christ by the power of his own Holy Spirit living inside of us. The power of his Word opens our minds and hearts to know truth from lies; light from darkness. We can practice knowing God's presence at all times. It does take practice, but why would anyone who has come to know the Lord not want to get as close with him as possible, despite our living conditions? We can live in God's heavenly kingdom, right here on earth. He says so. Remember the Lords Prayer? "Your kingdom come; your will be done . . . on earth as it is in heaven. Right here and right now.
John 16:22: And ye now therefore have sorrow: but I will see you again, and your heart shall rejoice, and your joy no man taketh from you.
23: And in that day ye shall ask me nothing. Verily, verily, I say unto you, Whatsoever ye shall ask the Father in my name, he will give it you.
24: Hitherto have ye asked nothing in my name: ask, and ye shall receive, that your joy may be full.
25: These things have I spoken unto you in proverbs: but the time cometh, when I shall no more speak unto you in proverbs, but I shall shew you plainly of the Father.
26: At that day ye shall ask in my name: and I say not unto you, that I will pray the Father for you:
27: For the Father himself loveth you, because ye have loved me, and have believed that I came out from God.
28: I came forth from the Father, and am come into the world: again, I leave the world, and go to the Father.
29: His disciples said unto him, Lo, now speakest thou plainly, and speakest no proverb.
30: Now are we sure that thou knowest all things, and needest not that any man should ask thee: by this we believe that thou camest forth from God.
31: Jesus answered them, Do ye now believe?
32: Behold, the hour cometh, yea, is now come, that ye shall be scattered, every man to his own, and shall leave me alone: and yet I am not alone, because the Father is with me.
33: These things I have spoken unto you, that in me ye might have peace. In the world ye shall have tribulation: but be of good cheer; I have overcome the world.


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- Wiper candidate Julius Mawathe trounced ODM's Irshad Sumra in the Embakasi South by-election
- The Kalonzo Musyoka party thanked supporters for choosing its candidate
- The party, however, cautioned Jubilee leaders against claiming to have played role in the victory
Wiper party has cautioned Jubilee party against taking credit for it's candidate, Julius Mawathe's victory in the just concluded Embakasi South parliamentary contest.
Mawathe floored Orange Democratic Movement (ODM)'s Irshad Sumra in a victory that attracted congratulatory messages across the political divide particularly from the ruling Jubilee party.
READ ALSO: Aden Duale gives up on Gender Bill, says women should fight for political space
READ ALSO: ODM floored in Ugenya, Embakasi South by-elections
In a statement seen by TUKO.co.ke, on Saturday, April 6, Wiper while appreciating the support of well wishers, expressed reservations over attempts by some leaders from the ruling party to closely associate with Mawathe's victory.
"We gladly appreciate the support of well-wishers towards the achievement of our victory but we urge some of them not to provoke us by trying to reap where they did not sow," said the umbrella party in the statement.
While the letter did not specifically mention Jubilee party, TUKO.co.ke understands that a section of leaders from the ruling party led by Deputy President William Ruto took to social media to celebrate Mawathe's victory even before the the Wiper leadership did so.
Ruto described the win alongside that of Ugenya's MP-elect, David Ochieng, as a reflection of the voice of the "hustlers", a group he often associates with.
Senate Minority leader Kipchumba Murkomen also celebrated Wiper's win and sarcastically questioned the ability of ODM to impeach the DP when it had lost popularly in its backyard.
READ ALSO: Wiper party ridicules ODM after humiliating Embakasi South by-election loss
Some of these reactions got the attention of Wiper party which felt some well wishers were going overboard to claim victory.
"This may be understood in the context of life in the wild, where a brave cheetah brings down an antelope but, just as he prepares to have his meal in peace, scavengers such as hyenas, vultures and ravens, circle in for the meal, claiming it as their own! the Kalonzo Musyoka lead outfit further took a jibe at Jubilee leaders.
The party further stated Kalonzo would vie for the presidency in 2022.
"Be that as it may, what has happened in Embakasi South is only a microcosm of what will happen in 2022 when our party leader, Kalonzo Musyoka, will throw his hat into the ring for the presidential race," stated Wiper.
Do you have a hot story or scandal you would like us to publish, please reach us through firstname.lastname@example.com or WhatsApp: 0732482690 and Telegram: Tuko news.
President Uhuru Kenyatta insists handshake is there to stay I Tuko TV


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Hair Stylists
Lisa Frassetto
Lisa is a highly educated and trained stylist with 16 years of experience. Her background consists of three Advanced Haircutting diplomas at the Vidal Sassoon Academy from London and California. She has various certificates at the Bumble and Bumble University in NYC that left her strong in advanced haircutting, razor cutting, specialty styling and up do styling. This education gave her the credentials to become an educator for the brand. Lisa also participated in Graham Webb's Platform of Art class and Vivienne Mackinder courses. These classes honed her skills in Bridal styling, vintage styling and the most up to date haircutting techniques. In addition, Lisa has attended multiple advanced color classes from Framesi, Redken, Tocco Magico, Aveda, and Artec. She also is certified in Great Lengths Hair Extensions.
Check out Lisa on her instagram for more hair transformations @Lisafrassettostyling
Tara Denninger
Tara is a highly educated and experienced stylist with over nine years in the industry.
Her extensive education includes Tocco Magico, Bumble and bumble, Italy Color, Brocato, Graham Webb, Redken, and TIGI.
Tara specializes in both Special Occasion and Wedding/Prom hairstyling.
She also is a TIGI educated makeup artist.
Check out Tara on her instagram for more hair transformations @TLDenningerHairStyles
Julez is a Redken Certified Hair Colorist!!
She attended Paul Mitchell The School, Jersey Shore, where she graduated in September of 2014.
She has a passion for cutting and specializes in transformation cuts! Julie loves to keep up with the most current trends in hair color and cuts; such as, Balayage, Ombres, Color Melts, Fashion Colors, Dip-dyes, Lobs, Pixies, Beach Waves, etc.
Check out Julie on her instagram for more hair transformations @hairbyjulez
Kelley Cover
Kelley is a Redken Certified Colorist and Hairtalk USA Extension Specialist.
She had attended multiple advanced color, cutting, and styling classes from Redken, Pureology, Hairtalk, Sam Villa, and more!
She is proficient in everything from color corrections, balayage colors, undercuts, fashion colors and razor cuts!
Kelley's work has been featured internationally in Look Magazine alongside Redken Platform Artist, Justin Isaac!
Check out Kelley's work on her instagram @hairbykelleycover
Tara Heffern
Tara has been a licensed Cosmetologist for over 11 years!
She graduated from Capri Institute, and continues to further her education at the Redken Exchange NYC!
She recently participated in Redken's "Cut & Know Why" and "Color & Know Why" courses!
Tara is a great Balayage artist and has a real passion for cutting!
She is also highly experienced and up-to date with the latest Bridal and Evening Hair and Makeup Trends!
Mya has been doing makeup for 5 years!
Mya is skilled in Bridal and Special Occasion makeup, as well as contouring, highlighting, and airbrush!
Whatever the occasion may be, Mya will make you look and feel amazing!
Most recently, Mya had decided to further her career, by enrolling in Cosmetology school. She will soon be expanding her services to hair, nails, and skin care!
Nikki Parsons
Priscilla has many years in the beauty business and specializes in special occasion and bridal make-up. She is also a dedicated hair stylist who is always looking to continue her education. Priscilla's education background consists of many advanced classes for cutting and coloring at the Redken 5th Avenue Academy in NYC, multiple classes in barbering and men's haircuts at the American Crew Academy, various classes at the Aveda Academy in NYC, and Rusk and Alterna courses as well. She is also certified in the Coppola Keratin Complex Straightening System.
Nikki Parsons is a Redken Certified Haircolorist and Haircolor Authority!
She specializes in the most current color trends like balayage, ombre, dip-dye, and other fashion colors.
Nikki is one of 500 caligraphy cut certified stylists in the US!
Most recently, Nikki won the 2019 License to Create award for "best processing picture" for hairpainting!
In 2015, Nikki's "Dip-Dye" color correction was published on ModernSalon.com. Her work was also featured on the front page of Redken.com and popsugar.com!
Nikki is certified in Wella Haircoloring, and has certificates in Haircutting and Haircoloring from the Redken Exchange on 5th Avenue in New York City.
She also has a certificate in Editorial Styling from Vivienne Mackinder. Nikki has an extensive knowledge of Redken haircolor and styling products. She is educated by Redken, Wella, Paul Mitchell, Sebastian, Alterna, Candy Shaw, & Living Proof. Nikki is also the salon's Living Proof Brand Enthusiast. She is constantly striving to improve her haircutting, coloring, and styling techniques.
Sally is a highly trained stylist with 17 years of experience. Her education background is extensive and includes Vidal Sassoon Academy, Bumble and bumble University, Great Lengths, Wella, Aveda, TIGI, Tocco Magico, Kevin Murphy, Sahag, and Paul Mitchell. She became a Network Educator for Bumble and bumble in 2003 and has been certified with Great Lengths since 2005. Sally specializes in many types of extension products, men's haircuts, all types of women's haircuts, bridal hair and makeup.
Check out more of Sally's work on her instagram @sallyostyling
Check out Nikki on her instagram for more hair transformations @hairsoakedinbleach
Peg Ohlsen
Since Peg became licensed in cosmetology in 2007 she has completed several additional courses in cutting, coloring, and advanced techniques with Tocco Magico, Bumble and bumble, Aveda, TIGI, Kevin Murphy, Redken and Coppola Keratin. Peg is also an educator with Tocco Magico color. Not only is she a skilled colorist, she is proficient in all types of cutting (including razored styles), foiling, special occasion styles, up-dos and makeup. Her specialties include thick hair, curly hair, and long hair.
Tonianne graduated from Ocean County Vocational Technical School in 2012. She specializes in Color Corrections and Balayage! She is also one of our in house makeup artists.
Tonianne is a hair and makeup educator at at The Ocean County Vocational Technical School in Brick, NJ!
Tonianne's work was published in both Modern Salon and Behind The Chair!
Check out more of Tonianne's work on her instagram @njglamourguru
Fran has been a stylist for 50 years!
She studied at the Martin Parsons School of High Fashion Up-dos's in Montreal Canada!
She has also continued her education with advanced hair cutting classes from Sam Villa, Redken, Sam Brocatto, The Vidal Sasson School NYC, and Bumble & Bumble!
Fran has a great passion for all phases of hair; coloring, cutting, and up-dos!
Nikki Renee
Check out Nikkis's work on her instagram @nikki_cutanddyeit
Nikki has been a licensed stylist for over 6 years!
She received her cosmetology license from The Capri Institute of Hair Design.
Nikki continues to further her education with the Redken Exchange NYC participating in courses such as Cut & Know Why and Color Trends!
She specializes in pixie cuts, transformation cuts, undercuts, men's cuts!
Nikki is very experienced and up to date with the latest color trends, such as Ombre color Balayage, Vivids, and Color Melts!
She is also a certified HairTalk Extension Artist!
Casey Bini
Check out Casey's work on her instagram @hairbycaseybini
Casey specializes in Redken Color and Design.
She has been a licensed cosmetologist since 2015.
Casey has furthered her education by taking various classes at the Redken Exchange in NYC, along with in-salon education!
Casey is also a HairTalk Certified Extension Artist and specializes in hair-cutting, balayage, color corrections, high-fashion vivid hair colors, special occasion styles, keratin treatments, and men's hair care and cuts!
Check out Nicolette's work on her instagram @Beauty_By_Nicolette
Stephen graduated from Paul Mitchell the School Jersey Shore in 2017. He is finally back behind the chair after taking a year away from Spa Bella to work on Wigs & Makeup for productions both on and off Broadway!
Stephen is also a certified makeup artist, as well as a Di Biase Extension Artist!
Michelle has been a stylist for 20 years! She is a Redken Certified Colorsit, who specializes in highlights, as well as 3D Balayage.
In addition to her Redken Certification, Michelle is also certified by Framesi for cutting and coloring!
Michelle is always continuing her education to offer the latest hair trends and corrective color techniques.
Janie has been working behind the chair for 10 years now! She has a passion for bringing out the beauty in each and everyone of her clients. She has attended multiple balayage, color/cutting, and styling classes from Kevin Murphy, Redken, Living Proof, Color Proof, Keratin Complex, Keune, and many more! She specializes in balayage and loves creating soft natural looks. She Is certified in ALL Keratin Complex Smoothing Treatments.
Janie has received a Certificate of Excellence with Color Proof and a haircutting certificate from the Redken Exchange in New York City. She looks forward to constantly continuing her education in and out of the salon to provide each client with the excellence they deserve.
Leslie has been a hairstylist for 12 years!
She graduated from Capri Institute of Hair Design and frequently attends education classes at the Redken Exchange NYC and The Art of Business in Reading, PA.
Leslie is passionate about working with different hair textures and enhancing an individual's natural hair, believing a thorough consultation every visit is most important.
She specializes in texture services, perming, men's hair design and barbering!
Leslie continues to further her education by attending numerous cutting, styling, and design courses with Pureology, Ruth Roche, Michelle Patton, Wendy Belanger, David Boyle, and Shannon Ring!
To achieve the best results, Leslie believes in always referring back to the principles of hair!


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The best part about Vegas is leaving. We made neither gains nor losses, because who can stand sitting in front of a pinball machine on speed and steroids anyway.
Desert, lots of it, and grey and monotonous through Nevada and the northwest corner of Arizona. Then Utah and it's all thick slabs of cheese and chocolate with a sprinkling of green in the valleys.
In Zion National Park, Utah, it's so hot setting up camp, especially pumping up the new blasted inflatable mattresses, I get dizzy.
The deer aren't too smart - they come right up to you.
We explore The Narrows, a deep crevice cut through the mountains with a river in it. Nothing like wading in cool water to freshen the whole body.
At night, a ranger talk on astronomy. We see Jupiter and its moons through a telescope. Conclusions: we're minuscule as a species, not even a single granule in the whole world of sand and dirt and dust combined. But we don't know it because we're polluting the sky with artificial light, making it harder and harder to see even the biggest, most obvious stars. Did you know, artificial light so corrodes melatonin and warps the circadian rhythm that it's carcinogenic? Goddammit, even that!


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I woke up at an unearthly hour today hence the pictures:
Vaio Sucky In-Built Camera plus flourescent table lights= Geisha without red lips/Ghost.
I did wake up at an earthly hour to go potty though and I could not get back to sleep even though I laid 'comatose' on my bed for almost two hours.
Oh well.
P.S Please do not stare too long at my face or you might just start imagining something in the black space behind me.


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But now I'm a bit addicted to them, which is something I never thought I'd say, what with being a ravenous carnivore and all (like a T-Rex from Jurassic Park, but less likely to eat Bob Peck - he's all gristle anyway, I've heard...) and I use them in all sorts of things, from being the basis of veggie chillies, lasagnes and Shepards pies (is it still a Shepard's Pie if there is no actual Shepard, umm, meat in it? Answers in the comments section please...)
This recipe is a little bit of a different take on the humble lentil soup, because we all like tyhings that are a bit different (and by different, I mean that this soup has a bit of vinegar to give it some zip)
This soup recipe is so simple, yet so filling, and shows how you can make any soup that bit thicker and more nourishing by the simple addition of lentils. And it's also wholesome, hearty and, as my Mother would say 'stick-to-your-ribs' food, which is great for these long, dark and miserable winter nights. Which is odd, as it's a Greek recipe if I'm not mistaken, where I assume that the nights are never cold and miserable, but I also assume that giant bronze statues stalk the land claiming sacrifices for the Gods, but that might just be Jason and The Argonauts, so you will have to excuse the mistake...
1 Potato
1 Large Onion
1 Large Carrot
2 Stalks of Celery
2 Cloves of Garlic
1l Vegetable Stock
250g Red lentils
50ml Tomato Puree
2tsp Chopped Oregano
1 Bay Leaf
100ml Red Wine Vinegar
Olive Oil
Salt and Pepper
1. Finely dice the potato, carrot, celery and onion. Heat some olive oil in a large pan, then add the chopped vegetables and the garlic. Cook the vegetables for about 5 minutes over a medium heat, until they start to become soft. Make sure to stir constantly or the vegetables can stick, and don't let them become too coloured or brown.
2. Add the stock, lentils, tomato puree, bay leaf and oregano then simmer the mixture for at least an hour until the lentils have cooked through and gone soft. Once this has happened, and the lentils are starting to fall apart in the soup, take the pan off the heat and allow the soup to cool down.
3. Transfer the soup to a food mixer and blend until smooth. Season to taste and add the red wine vinegar, then return the pan to the hob to heat it through, then serve with a swirl of olive oil.
4. Flat breads make an excellent accompaniment to this soup, which is so easy to make but so tasty. Enjoy!


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UNESCO declares Ahmedabad as world heritage city – July 2017
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UNESCO declares Ahmedabad as world
heritage city – July 2017
Ahmedabad, in western India, is the largest city in the state of Gujarat. The Sabarmati River runs through its center. On the western bank is the Gandhi Ashram at Sabarmati, which displays the spiritual leader's living quarters and artifacts. Across the river, the Calico Museum of Textiles, once a cloth merchant's mansion, has a significant collection of antique and modern fabrics.
UNESCO declares Ahmedabad as world heritage city – July 2017:
Ahmedabad has been declared as India's first world heritage city by United Nations Educational, Scientific and Cultural Organization (UNESCO).
Ahmedabad city has been declared as a World Heritage City at the 41st session of UNESCO's world heritage committee being held at Krakow in Poland. Nomination of Ahmedabad was supported by close to 20 countries, including Turkey, Lebanon, Tunisia, Portugal, Peru and Kazakhstan.
"Just inscribed as UNESCO world heritage site: Historic city of Ahmedabad #India", UNESCO said in a tweet on Saturday.
Gujarat Chief Minister Vijay Rupani and BJP president Amit Shah hailed the declaration of Ahmedabad as world heritage city by UNESCO, PTI reported.
Expressing delight at the announcement, Rupani said in a tweet, "Thrilled to learn that Ahmedabad has been recognised as UNESCO World heritage city, first of its kind in India"
"Delighted to know that UNESCO has declared Ahmedabad as World Heritage City. Proud moment for every Indian," the BJP national president said in a tweet.
Ahmedabad was in the race for the prestigious title along with Delhi and Mumbai.
The walled city of Ahmedabad believed to be founded by Ahmed Shah some six hundred years earlier has 26 ASI-protected structures, hundreds of 'pols' that capture the essence of community living and numerous sites associated with Mahatma Gandhi who lived here from 1915 to 1930.
In 1984, the first study for conserving heritage structures was carried out in the city. A heritage cell was also set up by the Ahmedabad Municipal Corporation (AMC).
The city had featured in UNESCO's tentative list of world heritage cities on March 31, 2011.
Gujarat :
Chief Minister : Vijay Rupani
Governor: Om Prakash Kohli
Capital: Gandhi Nagar
Read Also:
Vocab Master Day – 65


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In the current fast growing world the fashion leggings cheap are the very need of everyone. They are worn all around the world with different styles and shapes. They are comfortable and easy to wear and so making them famous worldwide. The teens are the ones who wear them almost with every outfit and they like the fashion leggings cheap really very much, and they are preferable for them on all other clothes. There are many styles in fashion leggings cheap, so it is easy to wear them with every outfit without any hesitation. The teens think that the fashion leggings cheap are the one to be worn with almost every outfit so they have many style tips to make the leggings comfortable with every outfit.
The most common trend in styling the leggings is the ripped leggings that are available as someone's desire and you can make them yourself as well. For this cause you must have the leggings in your desired color, a ruler to measure, the scissors which can cut the fabric easily, and a chalk or a pencil to mark the areas which are to be cut. The procedure is as under:
There are many more options and styles in the ripped leggings and they are in fashion nowadays as everyone wants change. Another option to style and rip the leggings is the easy one for those who do not want to do the long procedures and to do it with the go. For such people the steps are as follows:
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The Pseudomonas aeruginosa las (lasR-lasI) andrhl (rhlR-rhlI) quorum-sensing systems regulate the expression of several virulence factors, including elastase and rhamnolipid. P. aeruginosa strain PR1-E4 is alasR deletion mutant that contains a second, undefined mutation which allows production of elastase and rhamnolipid despite a nonfunctional las system. We have previously shown that this strain accomplishes this by increasing the expression of the autoinducer synthase gene rhlI. In this report, we show that the elastolytic phenotype of mutant PR1-E4 can be complemented with a P. aeruginosa homologue of the Escherichia coli dnaK mutation suppressor gene dksA. When supplied in trans on a multicopy plasmid, this gene completely suppressed elastase production by mutant PR1-E4. Cloning and Northern blot analysis revealed that dksA was neither mutated nor less transcribed in mutant PR1-E4. When overexpressed,dksA also reduced rhamnolipid production by both mutant PR1-E4 and the wild type, PAO1. Using Northern blot analysis andlacZ reporter fusions, we show that dksAinhibits rhlI, rhlAB, and lasB transcription. Exogenous N-butyryl–l-homoserine lactone overcame the reduced expression of rhlI and restoredrhlAB and lasB expression, as well as elastase production. Our results suggest that the overproduction of the P. aeruginosa DksA homologue inhibits quorum-sensing-dependent virulence factor production by downregulating the transcription of the autoinducer synthase gene rhlI.
Pseudomonas aeruginosa is a major opportunistic human pathogen. In P. aeruginosa the quorum-sensing circuitry, composed of the las and therhl quorum-sensing systems, regulates the expression of numerous genes, including lasB (elastase) andrhlAB (rhamnosyltransferase, required for rhamnolipid production) (35). The transcriptional activator LasR and the autoinducer molecule 3-oxo-C12-HSL [N-(3-oxododecanoyl)-l-homoserine lactone] constitute the las quorum-sensing system (21, 23). Similarly, the transcriptional activator RhlR and the autoinducer molecule C4-HSL (N-butyryl-l-homoserine lactone) constitute therhl system (18, 24). The lasI and the rhlI genes encode the autoinducer synthases that synthesize the autoinducer molecules 3-oxo-C12-HSL and C4-HSL, respectively. In cell-to-cell signaling, or quorum sensing, the concentration of the autoinducer molecule increases with bacterial cell density until a threshold concentration is reached. At this point the autoinducer binds to its corresponding transcriptional activator. The autoinducer-protein complex then activates the transcription of specific target genes (7, 10). Thelas and rhl quorum-sensing systems interact with each other, as the complex 3-oxo-C12-HSL–LasR activatesrhlR transcription (14, 26), and the complex C4-HSL–RhlR is necessary for optimal expression oflasB (3). P. aeruginosa strain PAO-R1 is a lasR deletion mutant (8) which is unable to produce elastase and rhamnolipid (36) and is significantly less virulent than the parent wild-type strain PAO1 (27, 33). We have previously described a strain PAO-R1-derived mutant, PR1-E4, which produces elastase and rhamnolipid despite the absence of a functional las quorum-sensing system (36). The precise site of the mutation in strain PR1-E4 is unknown; however, because of the extensive deletion in thelasR gene, a simple reversion is impossible. An increased expression of the autoinducer synthase encoding gene rhlIseems to compensate for the loss of the las quorum-sensing system in this strain.
To further characterize mutant PR1-E4, we complemented this strain with a wild-type P. aeruginosa gene bank and screened transformants for loss of elastase production. A complementing gene was identified and found to be a homologue of the Escherichia coli multicopy mutation suppressor gene dksA(13). In trans, dksA suppressed the elastase production of mutant PR1-E4 and reduced the rhamnolipid production of both PR1-E4 and PAO1. We demonstrate that overexpression ofdksA inhibits the expression of the autoinducer synthase gene rhlI, leading to a secondary reduction in the production of quorum-sensing-dependent virulence factors.
Media and culture conditions.Cultures were grown, with the appropriate antibiotics, at 37°C with shaking. PTSB medium (20) was used for P. aeruginosa cultures, and LB medium (28) was used for E. coli cultures. The defined, nitrogen-limited Guerra-Santos (GS) medium used for rhamnolipid determinations (11) was supplemented with 20% glycerol instead of 0.1 M glucose (17). Elastin-agar plates contained 0.5% elastin and 0.8% nutrient broth (20). For β-galactosidase (β-Gal) determinations, A medium (23) supplemented with 0.05% yeast extract, 0.4% glucose, and 1 mM MgSO4 was used. Antibiotics were used at the following concentrations when required: for P. aeruginosa, carbenicillin (200 μg/ml), tetracycline (100 or 50 μg/ml in solid or liquid medium, respectively), and gentamicin (100 μg/ml); for E. coli, ampicillin (100 μg/ml), tetracycline (20 μg/ml), and gentamicin (15 μg/ml). 3-oxo-C12-HSL and C4-HSL autoinducers were synthesized previously (22, 24).
Bacterial strains and plasmids.Bacterial strains and plasmids are listed in Table 1. TheP. aeruginosa gene bank, GB24, was kindly provided by U. Ochsner. This 95% complete gene bank contains a Sau3A partial digestion of the P. aeruginosa wild-type strain PAO1 genome, cloned into the BamHI site of the multicopy vector plasmid pUCP24 (30). It is composed of approximately 5,000 independent DNA fragments with a median size of 3.3 kb (2 to 7 kb) (U. Ochsner, personal communication). Plasmid pVD was constructed by cloning a dksA-containing 960-bp SmaI fragment from pVD99.3 into the Klenow-repaired XhoI site of pLPRI (36). Plasmid pDECP60 was constructed by cloning the 960-bp SmaI fragment of pVD99.3 into the SmaI site of pECP60 (26). The orientation of dksA in pDECP60 is opposite that of the adjacent rhlA′-lacZ fusion. Plasmid pPBL25 was constructed by ligating a Klenow enzyme-treated 3-kbEco0109 lasB′-lacZ-containing fragment obtained from pKDT37 (22) into the SmaI site of pUCP18. Plasmid pPBLD26 was obtained by ligating a 960-bp SmaIdksA-containing fragment of pVD99.3 into the Klenow enzyme-repaired XbaI site of pPBL25.
DNA techniques.We used standard techniques for DNA manipulations (28). Restriction endonucleases and DNA-modifying enzymes were purchased from Gibco/BRL or New England Biolabs. Plasmids were introduced into E. coli by transformation (28) and into P. aeruginosa by electroporation (31).
Cloning of the dksA gene from mutant PR1-E4.ABamHI-NotI digest of mutant PR1-E4 DNA was separated by gel electrophoresis and screened for the presence of thedksA gene, using a 32P-radiolabeled 350-bpSmaI-SphI probe obtained from pVD99.3. A 2,500-bpBamHI-NotI chromosomal DNA fragment of mutant PR1-E4 containing the dksA gene was cloned into theBamHI-NotI site of pBluescript SKII+. Colony blot hybridization on nylon membranes was performed using a 350-bp SmaI-SphI probe for dksAobtained from pVD99.3, using E. coli strain DH5α as a host. The dksA gene recovered from strain PR1-E4 was sequenced using a Li-Cor 4000L electrophoresis apparatus and Ladderman dideoxy sequencing kit.
RNA preparation and Northern blot analysis.Total cellular RNA was prepared as previously described (5). In brief,P. aeruginosa cells were grown at 37°C in PTSB medium to stationary phase (optical density at 660 nm [OD660] = 2.0 ± 0.2), when the las and rhl systems are active, collected, and lysed in 3.5% sodium dodecyl sulfate. RNA was recovered after centrifugation of cell lysates on a 5.7 M CsCl cushion and further purified by two phenol-chloroform extractions. Northern blot analyses were performed following standard protocols. After electrophoresis on a 2.2 M formaldehyde–1.2% agarose gel, RNA was transferred to Hybond nylon membranes and hybridized according to the Amersham protocol to 32P-labeled double-stranded DNA probes. DNA probes were obtained as follows. A 560-bp BamHI internal fragment of rhlA was obtained from pRL1500 (18), a 630-bp SalI internal fragment oflasB was obtained from pRB1801 (2), and a 250-bp KpnI-EcoRI internal fragment ofrhlI was obtained from pJPP41 (25). A 350-bpSmaI-SphI fragment from pVD99.3 was used as adksA probe. For Northern analysis of dksAexpression, total cellular RNA was obtained from PTSB cultures of strains PAO1, PAO-R1, and PR1-E4 in the early exponential phase of growth (OD660 = 0.8) and late stationary phase of growth (OD660 = 2.0). 32P labeling was performed by nick translation, and probes were separated from unincorporated nucleotides using NucTrap push columns (Stratagene). RNA experiments were performed twice with independent RNA preparations. To ascertain that the RNA of each strain was intact and loaded equally, we used a probe for the P. aeruginosa pilA gene (a 280-bpEcoRI-BamHI internal fragment of pilAfrom pPAO-2 [29]), which encodes the structural subunit of pilin. Previous studies have shown that pilA mRNA is not affected by the lasR deletion in strain PAO-R1 (9).
Elastase and rhamnolipid production assays.Elastase production was measured by elastin Congo red assays as previously described (25). Rhamnolipid concentration in P. aeruginosa culture fluids was determined as previously described by orcinol assays (25).
β-Gal activity assays.β-Gal activity was measured as previously described (16), with the following modifications. P. aeruginosa cultures were grown for 18 h at 37°C with vigorous shaking in PTSB medium supplemented with carbenicillin (200 μg/ml) and subcultured into the same medium to a starting OD660 of 0.15. Cultures were assayed for β-Gal activity at regular intervals during growth. Cells were washed twice and resuspended in A medium prior to β-Gal activity determinations. All experiments were done in triplicates and performed at least two times.
Determination of C4-HSL concentrations.Culture supernatants were extracted with ethyl acetate, and C4-HSL concentrations were determined in a previously described (25) bioassay using PAO-JP2(pECP61.5).
Nucleotide sequence accession number.The sequence ofdksA is accessible in GenBank (accession number AF062653 ).
Isolation of a P. aeruginosa dksA homologue that suppresses elastase production of mutant PR1-E4.We wanted to determine whether the mutation restoring elastase production of thelasR-deficient strain PR1-E4 could be complemented by a wild-type gene or suppressed by overexpression of a different gene. We therefore electroporated the wild-type P. aeruginosa gene bank GB24 into the elastase-producing mutant PR1-E4; 6,700 isolated clones were screened on elastin-agar plates for the absence of elastase production. We found one non-elastase-producing clone, PR1-E4(pVD99.0). Isolation of pVD99.0 revealed a 2.5-kb fragment from the wild-type PAO1 genome (Fig. 1). Neither plasmid pVD99.1, which contained a 1.98-kb fragment of pVD99.0, nor plasmid pVD99.4, which contained the other 523-bp fragment of pVD99.0, suppressed elastase production by mutant PR1-E4. Plasmid pVD99.3 was obtained during subcloning of the 2.5-kb fragment by ligating a 960-bp SmaI fragment of pVD99.0 into theSmaI site of pUCP24. This plasmid, when electroporated into PR1-E4, still completely suppressed its elastase production. Sequencing of the 960-bp insert of pVD99.3 revealed two complete putative open reading frames (ORFs), one from bases 292 to 735 (ORF1) and the second, in the opposite orientation, from bases 805 to 464 (ORF2). Subcloning a 520-bp SalI fragment of pVD99.0 created pVD99.5, which contains an intact ORF2 but lacks the first 153 bp of ORF1. Plasmid pVD99.5 did not suppress elastase production by mutant PR1-E4. This result, together with the finding that ORF2 exhibits codon usage unusual (38) for P. aeruginosa, makes it unlikely that ORF2 is responsible for the suppression of elastase production by mutant PR1-E4. In contrast, the 443-bp ORF1 had a codon usage typical for P. aeruginosa, and accordingly the Genetics Computer Group program CODON PREFERENCE (38) indicated a high coding probability throughout its entire sequence. A search of the GenBank-European Molecular Biology Laboratory sequence database revealed amino acid homology between the gene product of this ORF and the E. coli dnaK mutation suppressor protein DksA (76% identity and 87% similarity) (13). We had therefore identified a new P. aeruginosa gene, which was homologous to the E. coli gene dksA and suppressed elastase production by mutant PR1-E4.
Cloning and transcription levels of the dksA gene from mutant PR1-E4.To examine whether strain PR1-E4 bears a mutateddksA gene that had been complemented by the wild-typedksA gene on plasmid pVD99.3, we cloned the dksAgene from mutant PR1-E4. The dksA gene from mutant PR1-E4 was recovered and sequenced, and its nucleotide sequence was compared with that of the wild-type P. aeruginosa dksA gene as described in Materials and Methods. We found no mutation in either thedksA ORF or the 350 bp of DNA upstream from dksA(data not shown). To determine whether a reduction of dksAtranscription restored the elastase production of mutant PR1-E4, we also examined the expression of dksA in both exponential (early) and stationary (late) growth phases. Using Northern blot analysis, we could not detect a difference in dksA mRNA levels between the wild-type strain PAO1 and mutants PAO-R1 and PR1-E4 (Fig. 2). (Note that the difference seen for dksA between lanes 1 and 2 for late RNA is identical to that seen with the pilin probe and is therefore presumed to be caused by loading differences.) Consequently, the dskA gene in strain PR1-E4 does not contain a mutation, nor is its expression downregulated. The suppression of elastase production of mutant PR1-E4 by dksA supplied in trans is therefore a phenotypic complementation, similar to the mutation suppressor effect of the E. coli dksA gene on chaperone mutations (13).
dksA affects the production of virulence factors in both PR1-E4 and PAO1.To quantify the inhibition of elastase and rhamnolipid production by dksA, we performed elastin Congo red and orcinol assays in the presence of thedksA-containing plasmid pVD99.3 or the vector control pUCP24 (see Materials and Methods). dksA completely abolished the elastase production of mutant PR1-E4 (Fig.3A) and reduced its rhamnolipid production by 40% (Fig. 3B). We also wondered whether multiple copies of dksA could inhibit the elastase and/or rhamnolipid production of the wild-type strain PAO1. dksA only slightly reduced elastase production by strain PAO1 (Fig. 3A). However, the production of rhamnolipid was reduced by 35% when dksA was overexpressed in PAO1 (Fig. 3B). Therefore, dksA suppressed both elastase and rhamnolipid production in the absence of a functionallas quorum-sensing system but mainly affected the production of rhamnolipid in a wild-type background.
dksA inhibits the expression of genes belonging to the quorum-sensing circuitry.Expression of the elastase gene (lasB), the rhamnosyltransferase genes (rhlAB), and the rhl autoinducer synthase gene (rhlI) is partially restored in mutant PR1-E4 compared to its parent strain, PAO-R1 (36). To determine whether overexpression ofdksA affects the transcription of these genes, we performed Northern blot analysis of strains PAO1, PAO-R1, PR1-E4, and PR1-E4(pVD99.3), using lasB, rhlA, and rhlIprobes, as described in Materials and Methods. lasB, rhlA, and rhlI mRNAs gave intense signals in the wild-type strain PAO1, contrasting with mutant PAO-R1, in which the mRNAs of these three genes were barely visible (Fig. 4, lanes 1 and 2, respectively). All three mRNAs were detected in mutant PR1-E4 (Fig. 4, lane 3). These results confirm our previous report thatlasB, rhlA, and rhlI are barely transcribed in the absence of LasR in mutant PAO-R1 and that their expression is partially restored in mutant PR1-E4 (36). However, whendksA was supplied in trans on pVD99.3 to mutant PR1-E4, neither lasB, rhlA, nor rhlI message was detected (Fig. 4, lane 4). In contrast, the pilA message, used as a control, was not affected by the presence of dksA.
To confirm the suppressive effect of dksA on rhlImRNA levels, we measured the expression ofrhlI′-lacZ in the presence and absence of multiple copies of dksA, using plasmids pLPRI (rhlI′-lacZ) and pVD (rhlI′-lacZ dksA). The expression of rhlI′-lacZ in mutant PR1-E4 was reduced to 17% in the presence of dksA [PR1-E4(pLPRI), 18,201 ± 909 Miller units; PR1-E4(pVD), 3,135 ± 586 Miller units; measured at an OD660 of 1.8]. As PR1-E4 is an undefined mutant, we wanted to confirm the inhibition ofrhlI expression in a defined genetic background. We therefore compared the expression of rhlI′-lacZ from plasmid pLPRI and pVD in the defined ΔlasR mutant PAO-R1 and in the wild type, PAO1. rhlI is barely transcribed in mutant PAO-R1 because of a nonfunctional las quorum-sensing system (36). To compensate for this defect, we measured the expression of rhlI in mutant PAO-R1 in the presence of 10 μM C4-HSL. In both strains, the presence ofdksA reduced the expression of rhlI′-lacZ (Fig.5A). Therefore, dksAinterferes with the transcription of rhlI not only in the undefined mutant PR1-E4 but also in the ΔlasR mutant PAO-R1, despite constant C4-HSL levels, and in a wild-type background.
The effect of dksA on rhlAB andlasB transcription is indirect. dksA could primarily affect the expression of rhlI, leading to an indirect reduction of rhlAB and lasB expression. However, it could also directly inhibit the expression of these three genes. To address this question, we cloned dksA under its own promoter on plasmid pECP60, which contains a rhlA′-lacZreporter fusion, to obtain plasmid pDECP60 (see Materials and Methods). In wild-type strain PAO1, the presence of dksA intrans reduced the expression ofrhlA′-lacZ (Fig. 5B). These results confirm those of the orcinol assays that showed reduced rhamnolipid production in the presence of dksA (Fig. 3B). To determine whether the inhibition of rhlI transcription by dksA is responsible for this effect, we determined the effect ofdksA on rhlAB expression in the definedrhlI mutant PDO100 (3). As rhlAB is normally not transcribed in this mutant, these experiments had to be performed in the presence of 10 μM exogenous C4-HSL (26). These experimental conditions allow distinction of a direct effect of dksA on rhlAB expression from an indirect effect, due primarily to an inhibition of rhlIexpression. Indeed, if rhlAB transcription is not directly affected by dksA, then its expression should not be diminished in this strain, as the source of the C4-HSL autoinducer is exogenous. As shown in Fig. 5B, rhlABwas expressed similarly from plasmids pECP60 (rhlA′-lacZ) and pDECP60 (rhlA′-lacZ dksA), suggesting thatdksA does not directly affect rhlAB expression. To demonstrate that the inhibitory effect of dksA can be overcome by exogenous C4-HSL, we also determined the expression of rhlAB in the presence of exogenous C4-HSL in PAO1 and PR1-E4 (Table2). The addition of 10 μM exogenous C4-HSL restored the expression of rhlAB in both strains PAO1(pDECP60) and PR1-E4(pDECP60). These results show that exogenous C4-HSL can compensate for the inhibition ofrhlI expression by dksA and confirm the hypothesis that dksA does not affect rhlABexpression directly. The expression of rhlR is reduced to 18% of the wild-type level in both strains PAO1 and PR1-E4 (36) and is not further reduced in PR1-E4 by the addition of dksA in trans (assayed by Northern blotting [data not shown]). Apparently the concentration of RhlR is high enough to support, in the presence of an adequate C4-HSL concentration, the level of expression of rhlAB observed in mutant PR1-E4.
To determine whether dksA inhibits elastase production directly by inhibition of lasB expression, we used a similar strategy and cloned dksA under its own promoter on plasmid pPBL25, which contains a lasB′-lacZ reporter fusion to obtain plasmid pPBLD26 (see Materials and Methods). To avoid the interference of a reduced expression of rhlI, these experiments were also first performed in the rhlI mutant PDO100. Similar to the case for rhlAB, lasB expression is reduced in mutant PDO100 due to the lack of C4-HSL production (3). The transcription of lasB in mutant PDO100 was therefore measured in the presence of 10 μM exogenous C4-HSL. In these experimental conditions, the expression of lasB was not suppressed by dksA[PDO100(pPBL25), 16,640 ± 885 Miller units; PDO100(pPBLD26), 15,347 ± 697 Miller units; mean of five independent experiments ± standard errors, measured at an OD660of 2.0). To confirm these results, we determined the expression oflasB in mutant PR1-E4, using the reporter fusions pPBL25 and pPBLD26, in the absence and the presence of exogenous C4-HSL (Table 2). As expected, the expression oflasB was reduced in the presence of dksA. This inhibition was overcome by the addition of 10 μM exogenous C4-HSL. These results suggest that dksA does not inhibit lasB expression directly but reduces the production of the C4-HSL autoinducer, which leads indirectly to a reduction of las B and rhlAB expression. To support this hypothesis, we measured the C4-HSL concentrations in the culture supernatants obtained from the same experiments (Table 3). Not surprisingly, mutant PR1-E4 produced less C4-HSL autoinducer than PAO1, confirming the partial restoration of rhlI expression observed previously in this strain (36). The concentration of C4-HSL in supernatants of mutant PAO-R1 was at the limit of detection of our bioassay (the sensitivity of our bioassay was 0.04 μM). In the presence of dksA, no C4-HSL was detected by our bioassay in supernatants of strain PR1-E4. Therefore,dksA in trans severely reduced the production of the C4-HSL autoinducer, as expected from the inhibition ofrhlI expression by dksA. To finally confirm that exogenous C4-HSL can overcome the inhibitory effect ofdksA, we also measured the production of elastase by strain PR1-E4 in the presence of dksA and exogenous C4-HSL. As shown in Fig. 3A, the addition of 10 μM C4-HSL in elastin Congo red assays restored the production of elastase by strain PR1-E4(pVD99.3). It therefore appears thatdksA does not inhibit the expression of lasB andrhlAB directly but affects both elastase and rhamnolipid production indirectly by the inhibition of rhlItranscription, leading to reduced C4-HSL levels.
An upregulation of rhlI expression has been suggested to be responsible for the partial restoration of elastase and rhamnolipid production by the P. aeruginosa starvation mutant PR1-E4 (36). In the present study, we have complemented mutant PR1-E4 with a wild-type P. aeruginosagene bank carried on a multicopy plasmid. By screening for the loss of elastase production, we isolated a P. aeruginosa homologue to the E. coli dnaK multicopy suppressor genedksA. We have shown that dksA is neither mutated nor expressed at a reduced level in mutant PR1-E4. dksA intrans not only abolishes elastase production in mutant PR1-E4 but also downregulates rhamnolipid production in mutant PR1-E4 and in the wild type, PAO1. Using Northern blot analysis and plasmids carrying both dksA and rhlI′-lacZ, rhlA′-lacZ, and lasB′-lacZ reporter fusions, we have shown thatdksA reduces the expression of these three genes, as well as the production of the C4-HSL autoinducer. The addition of exogenous C4-HSL autoinducer compensates for the inhibitory effect of dksA and restores elastase production, as well asrhlAB and lasB transcription. These results suggest that overexpression of dksA inhibits rhlIexpression, leading to a secondary reduction of rhamnolipid and elastase production.
dksA homologues with strikingly conserved sequences have been isolated from E. coli (13),Haemophilus influenzae (4), Salmonella enterica serovar Typhimurium (34), and now P. aeruginosa. The cellular localization and function of DksA are still unknown. DksA shares amino acid similarity with the E. coli TraR protein, a putative transcriptional regulator (6). Both proteins contain a zinc finger domain (Cx2Cx17Cx2C) that facilitates binding to DNA. Interestingly, the putative zinc finger motif is the region most conserved between the known DksA homologues, suggesting that these proteins might also function as gene expression regulators. In E. coli, dksA expressed from multicopy plasmids suppresses the temperature-sensitive phenotype associated with deletion mutations in the heat shock genes dnaK, dnaJ, andgrpE280n (13) and in the mukB gene (41). It is also a multicopy suppressor of the conditional lethal phenotype associated with a prc null mutation inE. coli (1) and has been suggested to be involved in plasmid replication (19). The S. enterica serovar Typhimurium DksA homologue has been linked to virulence of this strain, as a ΔdksA S. enterica serovar Typhimurium mutant was impaired in the ability to colonize chickens (34). Both E. coli and S. entericaserovar Typhimurium dksA mutants show poor growth in minimal media and defects in glutamine-glutamate biosynthesis (13, 34). The S. enterica serovar TyphimuriumdksA mutant also yielded a higher RNA amount in stationary-phase cultures than the wild type (34). For these reasons, DksA has been suggested to be involved in stress responses such as the stringent response (34). Recently DksA has also been shown to be required for optimal translation of the stationary-phase sigma factor rpoS, as well as for the expression of several other genes in S. enterica serovar Typhimurium (37). DksA might therefore regulate the expression of genes which products are required for the stabilization of proteins during the stringent response and entrance into stationary phase (34)
The relationship between rpoS and the rhlquorum-sensing system in P. aeruginosa is a matter of debate. Whereas previous data suggested that the transcription ofrpoS might be regulated by the rhl system (14), recent experiments have suggested that RpoS might repress rhlI, rather than the rhl system influencing the expression of rpoS (39). Could an upregulation of RpoS, secondary to the overexpression ofdksA, explain the effects observed in this study? The repression of rhlI by RpoS manifests essentially during early exponential growth, whereas the inhibition of rhlI bydksA occurs mainly during stationary growth. Moreover, anrpoS mutation results in increased production of pyocanin, a secondary metabolite dependent on the rhl quorum-sensing system (whether this mutation also affects rhamnolipid production is unknown), but more importantly also in a 20% reduction in the production of elastase (32). This effect of RpoS on elastase production is opposite what would be needed to explain the inhibition of elastase production by dksA. It is therefore unlikely that the results presented in this study are due to an upregulation of RpoS alone. The overexpression of dksA could also affect another regulator of rhlI expression that has been previously suspected (14). This regulator could be the recently described Pseudomonas quinolone signal, which increases the expression of rhlI (15). This third P. aeruginosa cell-to-cell signal seems to acts as a link between the las and rhl quorum-sensing systems.
Our data show for the first time that DksA can interfere with quorum sensing. Further work is required to determine the physiologic function of DksA and whether it plays a significant role in the regulation of virulence factor production by P. aeruginosa.
We thank L. Passador and T. DeKievit for discussions, U. Ochsner for the gift of the GB24 gene bank, R. Comte for outstanding technical assistance, and L. Tabak, T. Barras, W. Kuhnert, and G. Campo for help with DNA sequencing.
This work was supported by NIH grant R01A133713-04 (to B.H.I.), NIH predoctoral training grant 5-T32 AI07362-09 (to J.P.P.), Cystic Fibrosis Foundation grant PESCI96FO (to E.C.P.), and a Wilmot Foundation grant and Swiss National Research Foundation grants 3231-051940.97 and 3200-052189.97 (to C.V.D.).
- Received 23 August 2000.
- Accepted 27 November 2000.
- Copyright © 2001 American Society for Microbiology


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Metadata Repository
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textures are a big part of hamilton, so that's where I've been pointing the lens the last few days. I was out shooting with my friend Paul when he lead me to some weeping willows down by confederation park. these old trees seemed to be sprouting bunyons everywhere. the term "gnarly, dude," crept into my head...
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Source Audio – Nailing Neil Young's Live Tone
Andy at Reverb.com Shows Us How to Nail Neil Young's Live Tone with Help from the True Spring Reverb
Here at Source Audio headquarters, we debate quite a bit about music and musicians, but there are a few topics on which we ALL agree. One is that Neil Young freakin' rules. So we were quite happy to see the True Spring Reverb make an appearance in our pal Andy Martin's Reverb.com video on "How Get Neil Young's Live Guitar Tone with 3 Boutique Pedals."
The True Spring Reverb is equipped with an engine inspired by the vintage Fender© 6G15 outboard spring unit that was a favorite of Neil's in the early Crazy Horse days. Check out the video as Andy romps through a few Neil Young classics like "Cinnamon Girl," "Powderfinger," and "Cortez the Killer." What do you think – did he achieve full Neil-ness?
This Month's Featured Neuro Presets
We could not be more fired up about the explosion of published C4 Synth User Presets. The growing legion of C4 owners is creating and publishing KILLER new presets every single day with the Neuro App. Here is the best news; for all of y'all that don't have the time or desire to jump into the Neuro Editor and start experimenting with preset creation, it is super-easy to use Neuro to explore, audition, and save any of these great sounds directly to your C4 (check out our Instructions for using the Neuro App with Mac, iOS, Android, and Windows videos for a quick tutorial). Below are some recent favorites…
Our friend Do Noise was one of the very first people to dive into the C4 Synth's Neuro Editor and use it to its full potential. DN fearlessly experimented with the C4's vast collection of sound features, from the dual programmable sequencers in his "Octave Sequenced Bass" preset, to the intelligent harmonization in "C7th Chords," to the alternate LFO wave shapes in "Dancing Filters." He also produced an excellent video demonstrating his presets. All of these sounds are super creative, fun, and may be just the thing you're looking for to add an unexpected twist to your latest recording.
Some excellent C4 Synth bass presets (that sound equally excellent on guitar) from our friend @Quatschmacher (a.k.a "Peter Kenney")! Quatschmacher has a grand collection of great presets with fantastic accompanying sound clips. Here are a couple of our favorites…
Introducing the Spectrum Intelligent Filter
…And just in case you missed it, last month we released the awesome new Spectrum Intelligent Filter. Source Audio originally made a name for itself with the Soundblox Bass Envelope Filter, and that rich tradition continues with our brand new, even more powerful and compact envelope filter. Check the link for some great demos of the Spectrum!