text: The crystallization of protein samples remains the most significant challenge in structure determination by X-ray crystallography. lattice dehydration on diffraction and (iv) the selection of high-quality crystal fragments for Rabbit Polyclonal to CG028 microseeding experiments to generate reproducibly larger sized crystals. Applications to X-ray free-electron laser (XFEL) and micro-electron diffraction (microED) experiments are also discussed. TEM-guided crystal growth). Thirdly, we establish the usefulness of high-quality crystal fragments from crystals or nanocrystals for the growth of reproducibly larger sized crystals. 2.?Materials and methods ? 2.1. Proteins purification, uV and crystallography fluorescence testing ? We used crystals produced from different protein examples, which were grouped into four different proteins classes: (i) soluble, (ii) membrane, SCH-503034 (iii) multi-protein complicated or (iv) viral. Sources for protein-purification crystallization and protocols circumstances are summarized in Desk 1 ?. Subsequent visible inspection of crystallization drops was attained using an Olympus SZX16 bright-field microscope. Granular aggregates, as previously described in Calero (2014 ?) and Stevenson, Makhov (2014 ?), and noticeable crystals which may be useful for nanoseeding tests had been assayed for UV tryptophan fluorescence using a Jansi UVEX microscope. UV-positive examples had been visualized using the program (Jan Scientific). Desk 1 Proteins purification and crystallographic circumstances 2.2. Microcrystal fragmentation ? Crystals had been washed with a complete level SCH-503034 of 10?l mom liquor to eliminate excess protein through the crystallization drop (that is strongly suggested to prevent the forming of bubbles during vortexing). Preliminary fragmentation of microcrystals utilizing a 3?mm Teflon ball (Hampton Analysis) and vortexing revealed minimal and highly abnormal crystal fragments (discover 3.1 and Supplementary Fig. S1for 30?s. After centrifugation, the answer was pipetted to resuspend crystal fragments and aspirated right into a clean 0 gently.5?l microfuge pipe for following experiments. 2?l from the crystalline option was evaluated using bright-field and UV microscopy to look for the performance of fragmentation (see 3.1). TEM evaluation of crystal fragments or crystal marketing previously more developed seeding strategies (DArcy software program (Jan Scientific). Manual quantification of crystal fragments was performed using the (Schneider (Kurt De Vos; http://rsb.info.nih.gov/ij/plugins/cell-counter.html) as well as the process for keeping track of crystal fragments followed regular cell-counting techniques. 2.5. Transmitting electron SCH-503034 microscopy (TEM) tests ? All crystals generated from the proteins listed in Table 1 SCH-503034 ? were subjected to analysis by negative-stain TEM. Approximately 5?l of nanoseeds was applied to 400 square mesh copper grids covered with a thin continuous carbon film (Electron Microscopy Sciences) and made hydrophilic by glow discharge (EmiTech) for 1?min at 25?mV under atmos-pheric conditions. Samples were incubated around the grid for 30?s before blotting and staining with a 2% uranyl acetate solution. Grids were mounted on a standard room-temperature specimen holder and inserted into an FEI Tecnai T12 microscope (FEI, Hillsboro, Oregon, USA) operating at 120?keV. Images were collected on a Gatan Ulstrascan 1000 CCD camera (Gatan, Pleasanton, California, USA) and FFTs were generated using the v.3.9.4 software (Gatan Software Team, Pleasanton, California, USA). Preparation of sample grids, data collection and evaluation of crystal images can be achieved in 2?h. 3.?Results ? 3.1. Crystal-fragmentation analysis using UV microscopy and TEM ? Generally, the crystal sizes resulting from crystallization screens are too large (Supplementary Fig. S1and 1 ? crystals showing partially ordered ( … For example, TEM analysis of crystals of the multi-protein complex comprising human damage-specific DNA-binding protein 1 (DDB1), DDB1-cullin 4 associated factor 1 (DCAF1), uracil DNA glycosylase (UNG2) and HIV Vpr (DDB1CDCAF1CUNG2CVpr) revealed high-quality lattices (Figs. 2 ? and 2 ? (Figs. 2 ? and 2 ? and 2 ? and Supplementary Figs. S5and S5and S3and S5and 4 ? and 4 ? and SCH-503034 dGTPase crystals. TEM analysis of pre-dehydrated and post-dehydrated crystals revealed improved lattice quality, as indicative by higher ordered, isotropic Bragg spots for both samples (Fig. 7 ?). In addition to improved lattice quality, higher resolution diffraction was also observed at the synchrotron for the dGTPase sample (Supplementary Fig. S6). Physique 6 Qualitative evaluation of the solvent content of crystal fragments. (sodium citrate, (magnesium acetate + 10% PEG 4000. Scale bars: 50?nm. The crystals in (and S7dehydration, the damaging effect of the stain itself and imaging at room temperature). Such behavior may explain the anisotropic diffraction observed for Pol II-GFP crystals in Fig. 4 ?. It is possible that the larger number of crystal contacts along the isotropic direction could withstand the effects of crystal dehydration while the direction with fewer contacts collapses. Conversely, we believe.


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Landscape Carpentry
Discover the beauty of a custom designed fence, gate, arbor or raised vegetable garden beds.
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The Garden Pros to Call From Spring to Fall
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Higher Ground Gardens was founded in 2008 by a husband and wife team. Together they have over 25 years of experience.
Our team at Higher Ground Gardens would love to hear from you!
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subset: commoncrawl


text:
From New Hampshire to North Carolina
R. M. Couch, a resident of Southern Pines for eight years, left his home in New Hampshire upon the advice of his physician. " Within five years there have been planted in this immediate section 1500 acres in fruit, and in order that your readers may have the advantage of direct correspondence with any or all the growers of fruit, I will give the names from memory: C. J. Eaglesfield was the pioneer on a small scale; S. N. Whipple, extensive peach, plum, grape and nut farm; Van Lindly Orchard Co., 350 acres peach, pear, plum and blackberry; Niagara Grape Co., 107 acres in grapes; Southern Pines Fruit-Growing Co., eighty acres in grapes; Benjamin Douglas, Jr., of Orange, N. J.; Tarbell & Carlton, H. P. Bilyeu, Dr. C. W. Weaver, C. D. Tarbell, Thomas Carlton, Fred Oberhouserheur, James H. Murray, S. W. Thomas, Charles H. Thompson, Edwin Newton, Doctors Boynton, Stevens and R. M. Couch, Rev. A. A. Newhall, B. Van Herff, J. T. Wilson, Dr. W. P. Swett, H. P. Stebbins, J. A. Morriss, R. S. Marks, L. S. Johnson, C. C. Mitchell, John Huttonhomer, F. J. Folley, Rev. J. W. Johnston, Mrs. L. A. Raymond, Mrs. Louisa Young, P. Pond, Fred Dixon and others. There were shipped from this point last season 150 tons, being the first bearing year of the oldest vineyards of much size. The bearing vineyards and orchards the coming season will more than double the shipments, and in two years all the vineyard trees mentioned will come to bearing. The prices in Washington and New York last July were six and seven cents per pound for black grapes, and thirteen and fourteen cents per pound for Delaware and Niagara, and $3.50 to $4.50 per bushel crate for peaches and plums. The demand was as good at the close of the season as at first. Write to Dr. C. W. Weaver, S. N. Whipple, H. P. Bilyeu, C. D. Tarbell, C. B. Mabore for prices obtained for their own shipments. Dr. Weaver realized from three acres of his best Delaware grapes $150 per acre net. Southern Pines is a town eight years old, in the midst of the turpentine region of North Carolina, sixty-eight miles southwest from Raleigh, on the Raleigh & Augusta Railroad (part of the Seaboard Air Line), fifteen hours from New York, and is six hundred feet above sea level, the highest point in the whole turpentine belt. The soil is a sandy loam and has a perfect drainage. Malaria is unknown. The presence of the long-leafed pine in large quantities causes the generation of ozone to such a degree as to make this locality almost a specific for throat and lung difficulties. Many physicians and a large number of the cured and benefited testify to its wonderful effects. The town is filled mainly with Northern people, and has four hotels, a good school, and church services every Sabbath. There are three stores, and railroad, telegraph and express offices. There are many fine residences and a large hotel 300 feet long and four stories is being built with modern improvements." more
Find your Ancestors Records on North Carolina Pioneers
Originally published on Blogger


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Joseph Chinenyeze Ibekwe is a leadership & entrepreneurship trainer; a Political Strategist; a Gender Specialist; a Development consultant; a life coach; a Journalist, Publishing Author and a Minister of the Gospel.
He is the founder/CEO, Foundation for Leadership & Education Development; a knowledge-based NGO established in 1995 to facilitate access to basic education and promote entrepreneurship and livelihood opportunities for women and youths. He is also established the Fled International Leadership Institute, an advanced professional Institute in 1999 to train and mentor leaders and technocrats in Nigeria and Africa. Through the NGO and the Institute, Joseph has trained about 5000 and mentored more than 2000 emerging leaders including women and youths in last 25 years.
He was formerly a development journalist with the Daily Times of Nigeria, Lagos where he reported extensively on issues of the environment, health, agriculture and Sciences in the 1990s. He is the Publisher of Future World Magazine; the convener of People & Power Conferences and Team Leader at Kingdom Impact Network (KIN) – an open membership of Christian leaders in Ministry, Politics, Business and Professions around the world. He is also the President and senior pastor at Chapel of Grace Transformation Ministry – a multi-dimensional ministry with emphasis on Church planting and preparing individuals for ministry.
In the past twenty five years, Joseph has developed leadership training programmes and conducted training sessions for professionals, women and youths around Nigeria. He is a 1996 Fellow of Salzburg Global Seminar, Austria and a 2010 CSO Fellow of the World Bank/IMF in Washington DC, USA. Between 2007 & 2008, Joseph served as Member, National Working Group (NWG) of the Africa Peer Review Mechanism (APRM) on the appointment of the Secretary to the Government of the Federation. He has authored several books including College Prefects are Leaders, Effective Writing for Career Excellence and Promoting Sustainable Human Development through Mentoring. His two recent books are The Quest for Political Leadership & Citizens' Democracy and Politics is Kingdom Business
He has been a consultant to UNESCO, Federal Ministry of Youth Development and Federal Ministry of Finance, Abuja in the areas of Strategic communications, organisational development, Strategic Planning, and gender mainstreaming. He is a trained UNESCO Development Expert Leader from UNESCO Training Centre, San Sebastian, Spain.
Joseph Ibekwe lived many years in Lagos and currently resides with his family in Abuja, Nigeria. He is married to Maryann Onyinyechi, JP, a senior civil servant and Fellow of Association of National Accountants (ANAN). He loves reading, writing, speaking and he plays golf.


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There's a hairy beast out on the prowl. It spots its prey from a distance, stalks along the forest floor, and pounces with a mighty flying leap. It then proceeds to suck its prey dry.
This animal is, of course, a tiny jumping spider.
You may not be used to thinking of spiders as strategic hunters, but jumping spiders have cat-like talents, bundled in package smaller than a dime. Take a look at how these spiders make your house cat look like an amateur hunter.


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Of average build, there is no use of brute force in Rohit Sharma's strokes. He is of the silken touch; the ones who depend on timing. That regal quality was on full display at Bangalore's Chinnaswamy Stadium on Saturday night. Sharma lit the stadium up with a show of six-hitting never seen before in a one-day game. Sending the holiday crowd into bouts of frenzy, the opener smashed a world-record 16 sixes during his hurricane double hundred. Powered by Sharma's super-show, India clinched the seventh one-dayer by 57 runs to win their first full series against Australia at home.It was not a convincing win though. Set a mammoth target of 384, James Faulkner had the Indian players' and supporters hearts in their mouths with a nerveless counter-attacking hundred.From 211 for eight, Faulkner single-handedly took the fight to the Men in Blue with his ninth-wicket partnership of 115 (89 balls) with Clint McKay. With 58 needed off 30 balls, Ravindra Jadeja got the breakthrough by clean-bowling McKay. Faulkner was out next ball, trying a big hit, for a valiant 116 (73 balls). Earlier, Sharma's day had not exactly started on a bright note. Having been involved in the run-out of the local IPL hero, Virat Kohli, for a duck, Sharma had upset the home supporters.An extraordinary effort was needed to win back the support. To his credit, he produced just that, more than compensating the fans with a show of scintillating strokeplay as he became the third batsman in history to score an ODI double hundred (209 off 156 balls).He surpassed Sachin Tendulkar's 200 not out and had Virender Sehwag's highest score of 219 also in his reach, but was out with three balls left of play.Sharma has been on record saying that he has the confidence in his ability to clear the ropes at will. It helps him pace his innings and that ability was to the fore here.Saturday was an innings of three distinct phases. He was content to play second fiddle when his opening partner Shikhar Dhawan. He shifted gears at the right time. His fifty came off 71 balls and after Dhawan's fall, he accelerated to 85 off 87. The progress from 90s was at a snail's pace with nine dot balls from 95 to 100. The Australians gave away only 22 runs in the batting powerplay (36 to 40 overs) to M S Dhoni and Sharma.It was a lull before the storm with the opener saving the real fireworks for the last. He went from 100 to 200 in 42 balls after his first hundred had come off 114.Sharma hit nine sixes in the last 10 overs as he just took off. Along with Dhoni, he went on to plunder 167 off 94 balls for the fifth wicket, 101 runs coming in the last five overs.The Australians had brought it upon themselves by dropping Sharma on 120. Had they got Sharma out even then, India might have struggled to post a total which would have tested them.ScorecardThe squads:India: Rohit Sharma, Shikhar Dhawan, Suresh Raina, Virat Kohli, Yuvraj Singh, Mahendra Singh Dhoni (Captain), Ravindra Jadeja, Ravichandran Ashwin, Bhuvneshwar Kumar, Vinay Kumar, Mohammed Shami.Australia: Aaron Finch, Phillip Hughes, Shane Watson, George Bailey (Captain), Adam Voges, Glenn Maxwell, Brad Haddin, James Faulkner, Nathan Coulter-Nile, Clint McKay, Xavier Doherty.
> >


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Yale Environmental Solutions Accelerator
Bringing together a diverse and forward-thinking community of students, professors, alumni, industry leaders, NGOs and beyond to participate in day-long events pushing the agenda forward on big sustainability challenges.
We are pleased to announce our inaugural Environmental Solutions Accelerator!
On October 7, 2017, Yale students will work in interdisciplinary teams to address the Future of Food in our own backyard through three special tracks:
- Food Access
- Food Waste
- Diets
This one-day event will serve as a foundation for pushing the "agenda on the urgent transition to more sustainable, secure, and equitable food and agriculture system." [1]
We have all faced our own challenges in the various stages on the food system, and this is the place to come together, share those challenges, and develop innovative solutions.
Questions? Contact us at firstname.lastname@example.com
[1] Global Alliance for the Future of Food
Ways to participate
-Students - apply to be part of a team and develop a sustainable solutions to current challenges in the food system!
- Applications have closed - thanks for the overwhelming response!
-Experts - become a mentor and help teams develop their solutions with your expertise
- For more info contact firstname.lastname@example.com
-Others - folks keenly interested in food solutions are encouraged to attend the presentations at the end of the day and hear the innovative ideas developed by Yale students. Let us know you are coming by registering here.
Event Logistics
When: 8:00am - 7:00pm October 7, 2017
Where: Evans Hall, 165 Whitney Avenue, New Haven, CT
Tentative agenda
Conference organizers have selected a specific topic with three tracks. Material on the topic will be provided to all applicants. Applicants are expected to have reviewed the material prior to arrival at the event.
Saturday, October 7, 2017
8:00-9:00 Team Breakfast, Check-in ends at 8:15
9:00-9:15 Welcome Remarks
9:15-9:30 Clients Introduced
9:30-9:50 Remarks on importance of collaboration across disciplines
9:50-10:00 Introduction to Ideation
- 12:30-2:30 Genius Bar - Teams aligned with experts, specialists, resources, and stakeholders. Teams are pre-registered to meet with mentors aligned with their subtracks but have access to all.
- 3:15 Presentations due
Participant Portal
Applications are now closed!
Sign up now for the Genius Bar
Sign up for a 15-minute slot. You'll have 5 minutes to present your progress to mentors, and they will provide 10 minutes of constructive feedback.
Genius bar begins at 12:30 PM and with last slot starting at 2:15 PM.
There are 16 slots so teams are encouraged to take at least one (2 max) time slots starting from 12:30PM with last slot at 2:15PM. This is on a first come first serve basis so please take advantage of this resource!
There will be mentors available outside of genius bars as well for additional mentorship.


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In 2020, mann Objecta started modernizing its IT infrastructure and integrating a powerful ERP & CRM system. The planning, production
[Read on …]
In 2020, mann Objecta started modernizing its IT infrastructure and integrating a powerful ERP & CRM system. The planning, production
[Read on …]
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It's been less than 24 hours since Thi'sl's latest album 'Beautiful Monster' was released, and already it's climbed it's way up the charts to place #5 in the iTunes Hip-Hop/Rap category. With 'Chronicles Of An X-Hustler' debuting at the #4 spot in the same category, where do you think Thi'sl's newest release will rank in the weeks to come?
The album is also #2 Top 100 Rap & Hip-Hop Paid Albums on Amazon.com.
Buy 'Beautiful Monster' on iTunes


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Southern Central Heat Wave October - November 2016
A high-pressure ridge that developed in late-October brought record-setting warmth to the southern and central United States. The unseasonal heat is expected to last through mid-November. One of the clearest findings of climate science is that global warming has already dramatically amplified the intensity, duration and frequency of extreme heat events. Due to global warming, the most extreme heat events now impact a global area 10 times greater than before. And the impact on moderate heat waves is also dramatic, with a seventy-five percent share of moderate heat events globally now attributed to climate change.
Fall heat breaks records across the southern and central US
A persistent upper-level ridge of high pressure developed in late-October 2016 and brought unseasonal heat to much of the central US.[1] Daily record highs have fallen in cities across Arizona, Utah, Colorado, Kansas, Texas, Louisiana, Mississippi, Alabama, Georgia, Virginia, West Virginia, Tennessee, Kentucky, Ohio, Missouri, and Wisconsin.[2]
On November 1 and 2, numerous cities set all-time highs for any day in November, including New Orleans, LA, Austin, TX, Jackson, KY, Huntsville, AL, Chattanooga, TN, Cincinnati, OH, and Milwaukee, WI.[3][4]
Phoenix set a record for the number of days in October that saw highs of 90°F or warmer, with October 30 marking the 26th day that the mercury topped 90 that month.[1] On October 27, Phoenix broke its record for the latest day experiencing a high temperature of 100°F by four days.[5]
Dozens of cities in the southern states of Kansas, Arkansas, North Carolina, Alabama, and Florida broke daily record highs on Monday, October 31, making it the hottest Halloween on record. Atlanta set a new record high on October 31 at 86°F, the latest day on record the city has experienced such heat.[2]
Denver finished the month of October with no measurable snow for the third-consecutive October, which is the first time this happened in Denver weather history.[6]
Extreme heat events, such as this heat wave, are the kind of weather events that increase the most as the climate warms. The more extreme the heat wave, the more likely it is due to the change in the climate. And global warming has led to a dramatic surge in the frequency of the most extreme heat events.
Global warming dramatically increases frequency of the most extreme heat events
A small shift in climate leads to a dramatic increase in the frequency of temperatures at the high end. The very most extreme events are the events most affected by climate change. As the average global temperature rises and the climate shifts, hot temperatures that were extreme under the old climate are closer to the middle of the new temperature range. Under the earth's climate system events closer to the midpoint of the climate range occur much more frequently than events closer to the extremes, as shown in the graphic on the right. The shifting bell curve also leads to the occurrence of never-before-seen extremes in high temperatures.[7][8][9]
"...it is the rarest and the most extreme events - and thereby the ones with typically the highest socio-economic impacts - for which the largest fraction is due to human-induced greenhouse gas emissions." [10]
Due to global warming, the most extreme heat events now impact a global area 10 times greater than in the period 1951-1980.[7]
The impact on moderate heat waves is also dramatic, with a seventy-five percent share of moderate heat events now attributed to climate change.[10]
Many urban areas across the globe have witnessed a significant increase in the number of heat waves, with the largest number of heat waves occurring in the most recent decade studied, 2003-2012.[11]
Heat waves have generally become more frequent across the US in recent decades, with western regions setting records for numbers of these events in the 2000s. Recent multi-month extreme heat events in the US are unprecedented since the start of reliable instrumental records in 1895. There has also been a dramatic increase in nighttime temperatures in the US, reducing the number of critically important relief windows during heat waves.[12]


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He heals wounds of every shattered heart. Proverbs 147:3
Happy Tuesday. Today, I would like to ask you to join me in trying something new . . . stomp out childhood sexual abuse!
Imagine you are a child and a trusted family member does unspeakable acts to you that you do not understand. One of the people you love most in the world is not only aware, but jealous. You hide and lock the door, and he apologizes and promises to never do it again, but over and over he returns pushing the envelope further. You tell your mother, who scolds you and you keep the secret.
Imagine yourself a young adult with small children still afraid to be left alone with the perpetrator. A child in your family tries to kill herself from depression. She later tells similar stories about the same family member.
Imagine you are going through life and through no fault of your own, your world is turned upside down and inside out. All the people you know and love suddenly love you no longer. You are alone. Doing what's right to protect a child who has experienced a pain like your own means losing everything. You have nothing to gain. You are looked upon as dirty and substandard.
Imagine wanting to die from the pain of being isolated, unloved and invalidated. Each day the torment of memories is eating away at your existence. You can barely leave the house for fear of being known, ridiculed or accidentally seeing the people who hurt you.
Imagine the church that you attend supporting the people who hurt you without ever speaking with you, even though you were a victim and also a member. You know secrets unveiled from within the church walls and when you try to speak with the minister who had a hand in getting the one who hurt you most out of jail, he threatened to call the police.
Imagine the child you helped recanting everything as an adult, leaving you looking like a liar. You never again have any substantial relationship with anyone outside your husband and children.
Imagine working your way through college and fighting your way into a career later in the life. You fight to protect your children from people and situations that might be harmful, so they do not know the same pain.
Imagine yourself working through the pain and forgiving the ones who hurt you. You trust God and make a new life for your family and you are finally finding peace. No one can hurt you or your children now. Many years have passed. The family is forever broken and there is no hope of reconciliation. You now realize that we are all God's children and wish no one any harm.
Imagine yourself seventeen years later. The family member you loved so much has died. She never made things right with you. The same church places a plaque up in her honor, even though she hid family secrets about your pain. You seek out family history and convince yourself that she made stupid mistakes as a young adult and lived in a different time. You tell yourself she made decisions because women married to ensure they had providers for their children. You are able to reestablish a relationship with one relative who is also outcast by the family.
Imagine yourself twenty-four years after the arrest. The perpetrator has died. The family is fighting and excluding others as their greed takes control of their minds and hearts.
Imagine praying for the family and repose of the soul of the perpetrator. Imagine feeling numb at first and then sadness because your dreams that one day your family would be reunited and know you told the truth will never happen.
Someone I care about was a victim of childhood sexual abuse and experienced these events and feelings. If you know someone like this, whether a child or adult, don't look at them as dirty. Give them love and help them. It's never a child's fault. Childhood sexual abuse is real and destroys lives and families. Often these crimes are committed by people close to the family, if not the family. Children do not want their family members to be mad at them and more often than not, remain silent. Perpetrators need help too. Protecting the perpetrators only hurts more people. Churches are made up of people like you and me. We need to unite to help.
Join me today in stopping sexual abuse of children. If you have been abused, I lift you up in prayer. Leave me your name and I will pray for you. Join me in turning everything over to the one who mends shattered hearts. God bless you.