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Guide to the W. Bryan Bolich papers, 1891-1972
Series: Personal, 1861-1969
Series: Duke Law School, 1940-1966
Series: Professional activities, 1928-1973
Series: Writings
Series: Clippings, 1917-1966
W. Bryan Bolich (1896-1977) served as a Professor of Law at Duke University from 1927 to 1966. Papers contain family memorabilia, general correspondence, photographs, an oral history, diaries, course notes, writings, drafts of statutes revisions, and clippings. Major subjects include family work at Southern Railway in Forsyth County, N.C., Duke Law School curriculum development and reorganization, Law Day, the Rhodes Scholarship, Trinity College Class of 1917 alumni activities, Law School Alumni Association, North Carolina House of Representatives, and property and alien rights laws authored with the North Carolina General Statute Commission.
W. Bryan Bolich papers
Bolich, W. Bryan, 1896-1977
5 Linear Feet, 5,000 Items
Personal, 1861-1969
Duke Law School, 1940-1966
Professional activities, 1928-1973
Clippings, 1917-1966
Correspondence, manuscripts, photographs, diaries, certificates, correspondence and drafts of laws, miscellaneous memorabilia from Bolich's career as a Professor at Duke University Law School, student notebooks from courses taken at Trinity College and University of Oxford, and a taped memoir and 19-page transcription. Family materials include materials from Southern Railway and much correspondence between Bolich's parents during their courtship (ca 1891-1893). Political correspondence between Bolich and Richard Nixon is filed in Series 3 (Correspondence. Political).
In off-site storage; 48 hours advance notice is required for use.
[Identification of item], W. Bryan Bolich Papers, Duke University Archives, David M. Rubenstein Rare Book & Manuscript Library, Duke University.
1. Personal, 1861-1969
Contains general correspondence between W. Bryan Bolich and his family (bulk 1915-1919), friends, and colleagues (1920-1960). Earlier correspondence (bulk 1891-1893) was primarily between Sallie McMahon and John Alonzo Bolich (parents of W. Bryan Bolich). Family items include miscellaneous memorabilia and items from father John Alonzo Bolich's work as a train master at Southern Railway (ca 1907-1940). Also contains Trinity College class notebooks (ca 1917), Trinity College Class of 1917 Alumni Association materials (1942-1966), and University of Oxford notebooks (ca 1921-1927). The oral history (1969) contains a tape and transcript. Ordered by date.
1891-June 1892
July 1892-December 1892
January 1893-March 1893
May 1893-December 1893
1916-May 1917
June 1917-December 1917
January 1918-July 1918
August 1918-December 1918
Southern Railroad
The Floods of July, 1916
Class of 1917 (1942-1966) (Folder 1 of 3)
Class notes, 1917
Diaries, 1921
Diaries, circa 1921-1924
Class notes, 1921-1924
Transcript, 1969
Tape, 1969
2. Duke Law School, 1940-1966
Contains general correspondence with colleagues at Duke regarding Duke Law School curriculum development and reorganization, establishment of Law Day, activities of the Law School Alumni Association, and the 1963 building dedication. Also contains sabbatical and other leave requested by Bolich, course reading lists, drafts of "Duke Law School, 1868-1968: A Sketch" and sketches submitted by Bolich to The Bar Rag, official newspaper of the Duke Bar Association. Ordered by topic.
Alumni Association. Miscellaneous, 1940-1958
Alumni Association. Mordecai Portrait Fund, circa 1930-1932
The Bar Rag, 1950-1965
Building Dedication, 1962-1963
Curriculum, 1940-1941
Curriculum, 1965
Duke Law School, 1868-1968: A Sketch
Justice Jackson's Honorary Degree
Law Day, 1961-1964
Reorganization, 1933-1959
Retirement, 1966
Sabbatical and other leave
3. Professional activities, 1928-1973
Contains miscellaneous correspondence between Bolich and colleagues regarding research, conferences, general curriculum, laws, and politics. Also contains items from the Rhodes Scholarship Committee at Duke University (1953-1965). The political correspondence is primarily between Bolich and persons in the White House (including Nixon). Ordered by date.
Correspondence. General. 1928-1973
Correspondence. Political. 1933-1966
Resumes and honors, 1928-1965
Rhodes Scholarship Committee, 1953-1965
4. Writings
Includes Cases and Materials manuscript materials, "Constructing Wills", articles on landlord and tenant laws, correspondence and drafts of laws regarding property rights, short miscellaneous and creative writings, and text of speeches. Ordered by topic.
Cases and Materials, pages 1-175
Cases and Materials, pages 176-336
Constructing Wills, 1965-1967
History of the North Carolina General Court, 1948-1963
Landlord and Tenant articles. Correspondence, 1969-1970
Landlord and Tenant articles. Drafts, 1969-1970
North Carolina Bar Association. Mineral rights, 1959
North Carolina Constitution. Acts Barring Property Rights (Chapter 31A), 1960-1965
North Carolina Constitution. Devolution of Devises (GS31-42), 1965
North Carolina Constitution. Estates of Missing Persons (Chapter 28A-8), 1961-1965
North Carolina Constitution. Fiduciary Powers, 1965
North Carolina Constitution. General Statutes Commission. Miscellaneous Correspondence, 1957-1965
North Carolina Constitution. Interstate Succession Act (Article X, Section XI), circa 1960
North Carolina Constitution. Interstate Succession Act (Article X, Section XI). Correspondence, 1959-1964
North Carolina Constitution. Justice of the Peace, 1936-1943
Rule in Shelley's Case, 1941
Short works
Speeches, circa 1933-1954
5. Clippings, 1917-1966
Contains clippings about coaching sports at Trinity Park School, winning the Rhodes Scholarship, running for a seat in the North Carolina House of Representatives, becoming a professor of law, fraternity inductions, and general commentary on developing laws. Ordered by date.
W. Bryan Bolich was born in 1896 in Salisbury, North Carolina. He began his education at Wofford College (Spartanburg, S.C.), then transferred during his junior year to Trinity College (A.B. 1917) where he studied English and Economics. After graduation from Trinity College, he taught classes and coached baseball and basketball teams for the Trinity Park School in Durham, N.C. In 1918, Bolich enlisted in the U.S. Navy. He quickly rose from Seaman to Chief Petty officer and was later commissioned as an Ensign. He served for two years in World War I (1918-1920) and then returned to Trinity College to begin his law education (1921-1922). In 1922 Bolich was awarded a Rhodes Scholarship and attended Oxford University where he earned his law degree (B.A. 1923, B.C.L. 1924, M.A. 1927).
As a student he was a member and officer of many fraternities, including the "9019" honorary scholastic fraternity of Trinity College, Phi Beta Kappa (a scholastic fraternity), President of Order of the Coif (an honorary legal scholarship fraternity), Omicron Delta Kappa (an honorary leadership fraternity), the Tombs (an honorary athletic fraternity), Chief Alumnus of Kappa Alpha (a social fraternity), and Phi Delta Phi (a professional legal fraternity). He was also the 1917 Class President of Trinity College and was active in raising funds and organizing class reunions.
Upon his return from Oxford in 1927, Bolich was admitted to the North Carolina Bar Association and moved back to his hometown of Winston-Salem, N.C., where he practiced Civil and Criminal Law in state and federal court. In 1927, he was elected to the North Carolina General Assembly and served one term as the democratic representative of Forsyth County, N.C.. There Bolich drafted and introduced one of the first Alien Rights Registration Acts adopted in the United States. Later in his career, as a member of the North Carolina General Statutes Commission, Bolich was instrumental in preparing the amendment to the North Carolina Constitution which equalized property rights of married couples. He also helped write amendments and propose new laws on inheritance, estate rights, and landlord and tenant relations.
In fall 1927 Bolich returned to Duke University to teach legal history, real property, procedure, and landlord and tenant relations at the Law School. As a Professor, he was a member of the Duke University National Council, Duke Law Alumni Association Council, the American Association of University Professors, and served as a Rhodes Scholarships Institutional Representative for Duke University. He was instrumental in revising the Law School's curriculum and successfully argued for a required third year of study. He also established the Duke Law Alumni Association and edited the school's first alumni directory in 1935. He assembled a "Law Day" program that brought in panels of influential speakers including Duke Law alumni. Bolich was a professor to Duke Law alumnus Richard Nixon. During 1957, Bolich was a Visiting Professor at the University of Texas, Houston.
Bolich was the author of Activities of the North Carolina Bar Association, 1925-1935 (1936), Cases and Materials on Introduction to Procedure (undated), and Duke Law School: the First Hundred Years (1968). After his 1966 retirement from Duke University Law School, Bolich moved to St. Petersburg, Florida, where he lived until his death in 1977.
Faculty Collection (School of Law Library, Duke University)
Law Archives (School of Law Library, Duke University)
Duke University. School of Law -- Alumni and alumnae
Duke University. School of Law
Duke University. School of Law -- Study and teaching
Duke University. School of Law -- Students
Duke University. School of Law -- Faculty
North Carolina. General Statutes Commission
North Carolina. General Assembly. House of Representatives
Nixon, Richard M. (Richard Milhous), 1913-1994 -- Correspondence
Southern Railway (U.S.)
Trinity College (Durham, N.C.) -- Students
The W. Bryan Bolich papers were received by the University Archives as a transfer in 1969-1975.
Processed by Emily Glenn, November 2002
Encoded by Kimberly Sims, September 2006
Accessions A69-54, A72-33, A75-50, A71-387 were merged into one collection, described in this finding aid.


Enhancing community engagement, awareness and strengthening partnerships.
The expertise, experience, and enthusiasm of Bangor residents for the community and region will help make the workshop a success. We look forward to seeing you there.
Who: Stakeholders and community leaders including local residents and elected officials.
What: Discussions on health and community livability, group work sessions, and networking that will result in a Next Steps Action Plan.


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Texas Self-Defense: Police: Woman shot local man in the face as he choked her :: 09/12/2018
WACO, Texas (KWTX) Police are investigating after they say a woman shot a man in the face as he choked her during an argument at a Waco apartment complex.
Waco police confirmed Wednesday that officers were sent to the Arlington Farms apartments in the 1800 block of Primrose Drive just before midnight.
When officers arrived they found a man shot in the face.
Police later identified the man as Homer Hampton, 40.
During the investigation officers discovered Hampton and woman, who are in some sort of relationship or possibly related, were fighting when the man began to choke the woman, Waco police Sgt. Patrick Swanton said.
During the struggle, Hampton pulled out a gun, but the woman was able to point it toward him and shoot him, Swanton said.
He was taken to a local hospital.
Information about his condition wasn't available.
Swanton said charges against Hampton were pending and that charges against the woman were not expected because she fired in self-defense.
http://www.kwtx.com/content/news/Woman-shoots-man-who-was-strangling-her-police-say-493060901.html?ref=901

redpajama_set_name: RedPajamaCommonCrawl


If you think about planning for retirement, you wouldn't think it applies to you if you're still in your late 20s or early 30s. But the fact is the earlier you plan your retirement and start saving, the better it is for you to have a happy retirement. A lot of people in their 20s and early 30s believe they are not ready for retirement and they always leave it for later. If you only start retirement planning later on in your 40s, you may find it extremely hard to save up for your own retirement if you still have debts to repay.
If you have accumulated debts which haven't been fully repaid yet by the time you reach retirement age, it makes it more difficult to transition to retirement. Most people think they will have finished paying their debts by the time they reach that age but only if they were disciplined in saving and spending, will they be able to be free from debt. And it's not easy to repay debts while on pension since you no longer have any regular pay checks coming in.
Being financially unprepared before your retiring years, can force you to keep working longer to claim an Age Pension or any plan that's offered at your time. Investing can be a great source of income if you do it wisely; if done well, it can give you the option to retire whenever you want.
The best method for retirement planning is to start as early as possible. Setting a plan to save a few dollars from every pay check you get and actually sticking to the plan can be highly beneficial in your financial plans. Even from the first pay check you ever receive, put aside a few dollars like $5 a week or $50 a month can be great for your retirement fund. You might not notice it if you only put aside a small amount each week or each month but after a few months or even a few years, you'll notice you've already saved up a lot of money.
The transition to retirement will not be difficult if you create a retirement plan and stick to it. If you're savvy with prospective investments, they can also help make your transition easier. Superannuation funds are there to help Australians have a comfortable retirement but if you haven't reached the age to access your superannuation yet, your savings and investments can be very helpful.
In order to have an indication of how much you would need to retire happily, you will need to find out if the return you're getting from your investments can cover your current living expenses. If you are a savvy investor, you may be able to retire before you even reach retirement age. An easy way to calculate how much money you need to retire is to work out how much you actually spend, double that number and add a zero at the end. This amount will indicate how much you need you would need to retire comfortably. However, this indication may only work if your investments have regular returns and if your superannuation covers it.
If there's a significant gap between your figures, you might have to consider saving more over the years. If you still have debts, you should clear them quickly so your retirement planning can come into effect quicker.


Seven new books have been released by CDDRL affiliated faculty members Joshua Cohen, James Fishkin, Michael McFaul, Kathryn Stoner-Weiss, Joshua Teitelbaum, and Jeremy Weinstein.
%people1% - The celebration in Prague should have been more raucous. The most successful alliance in world history has extended to corners of Europe unimaginable just a few years ago.
Ambassador John Beyrle presented his credentials to President Parvanov as U.S. Ambassador to Bulgaria on September 8, 2005.


EisnerAmper announced that Andrew C. Bernstein, CPA/CFF, CVA, has joined the firm as a Managing Director in its Forensic, Litigation & Valuation Services (FLVS) practice. In this new role, Bernstein will work from the Miami and Fort Lauderdale offices.
Bernstein has more than 30 years of experience providing expert testimony on economic damages, valuation and business issues in complex business litigation. He has testified in Federal District Court, Federal Bankruptcy Court, State Court, Delaware Chancery Court, International Arbitration, and other venues.
Bernstein has led forensic accounting investigations of banks, health care entities, public companies, governmental entities and closely held companies in a wide range of matters including responding to regulatory actions, shareholder claims, concerns of financial fraud and internal corporate concerns.
Bernstein has been appointed as a receiver in Federal Court; an arbitrator by a State Court Judge and served as ICC Arbitral Panel member. As accountant to receiver or special master, he has presented reports and testimony in Federal and State courts.


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Questions raised over arrest of Mexican national in Missouri
by: MARGARET STAFFORD, Associated Press
KANSAS CITY, Mo. (AP) — Federal immigration officers trying to arrest a Mexican national smashed a car window and dragged him from the vehicle in front of his girlfriend and two young children, prompting condemnation from the woman's Missouri congressman.
A Facebook Live video taken during the arrest of Florencio Millan-Vazquez in Kansas City on Monday shows Immigration and Custom Enforcement and Kansas City police officers trying to persuade him to leave the vehicle. Millan-Vazquez, who is in the U.S. illegally, and his girlfriend, Cheyenne Hoyt, repeatedly ask to see a warrant. After officers warn that they plan to break the window, an immigration officer smashes the glass and others help drag Millan-Vazquez out of the car.
"I'm still in shock," a crying Hoyt told The Associated Press Tuesday. "You think that it's not going to happen to my family, like I never thought this was going to happen. You hear the things (President Donald) Trump says, but (Millan-Vazquez) is not a rapist, he's not a murderer, or a drug dealer. And the way they did it in front of the kids, they didn't care."
Hoyt, who shot and posted the video, said they had been on the way to a doctor's appointment for their disabled 7-month-old daughter when immigration officers blocked them in at their apartment complex and ordered Millan-Vazquez out of the vehicle. The immigration officer asked several times to see proof that Millan-Vazquez is allowed to be in the U.S. and tells the couple that ICE doesn't need a warrant to arrest someone who has entered the country illegally.
Agents must have a warrant when arresting someone inside a home, but the law is less clear for arrests outside a residence, said Jessica Piedra, an immigration attorney with the Kansas City Metro Immigration Alliance.
"It's a little murky since he was in his car," Piedra told The Kansas City Star.
Hoyt acknowledged Millan-Vazquez was in the country illegally but said he is a hard-working chef and family man who never caused any trouble. Immigration officials said Millan-Vazquez re-entered the country twice after being voluntarily deported in 2011 and that he has misdemeanors on his record.
"Millan-Vazquez was uncooperative and refused to exit his vehicle or follow lawfully issued commands issued by ICE and local police," said ICE spokesman Shawn Neudauer. "After attempting to negotiate with Millan-Vazquez for about 25 minutes, the ICE officers were left with no other choice than make the arrest by physically removing him from the vehicle."
On the video, the couple's 11-year-old son is heard crying as the window is broken and his father is taken away. Millan-Vazquez can later be heard asking to say goodbye to his son. An ICE agent turns down his request.
"Right now we're being extremely nice to you, but what you just put us through, what we had to go through, you're lucky I'm letting you talk to (Hoyt) right now," the agent says. "So no, you can talk to him later, she can bring him up to where you're gonna be and you can see them there."
The arrest occurred on the same day the Trump administration announced that it was extending the authority of immigration officers to quickly deport immigrants who have been in the U.S. illegally for less than two years without putting them before a judge.
It also came the same day that an immigration official in Nashville, Tennessee, gave up trying to arrest a man who refused to leave his vehicle with his 12-year-old son. Neighbors surrounded the van, bringing the man and his son water, food and gas for the vehicle.
U.S. Rep. Emmanuel Cleaver of Missouri called the video of Millan-Vazquez's arrest "heartbreaking and gut-wrenching."
"It should make every American question where we're headed as a nation and what kind of people we want to be," Cleaver said in a statement. "This President has spent the entirety of his term demonizing immigrants and fearmongering his base into relinquishing the very values that have made our nation what it is today, and this incident is a direct result of that rhetoric."
Cleaver, who first posted about the arrest on Twitter on Monday, said he had communicated with the regional office of ICE and will continue to monitor the situation, adding that the only way to ensure such incidents don't happen again is for Congress to pass bipartisan, comprehensive immigration reform.
Several immigration advocates said during a news conference Tuesday that Kansas City police damaged the public's trust by helping ICE with the arrest. Police spokesman Jacob Becchina said officers frequently back up federal, state or other officers who conducting enforcement in Kansas City.


1.10 New BL21 cells ready to use!
10.2 New XL1b and BL21 cells ready to use!
Results:The expected bands should run at 1700 and 3000 bp. P679 was sent for sequencing.
Results:The expected bands should run at 1700 and 3000 bp. P684 was sent for sequencing.
Results:The expected bands should run at 500 and 4500 bp. However, just one band at ~3000 bp is visible on the gel.
I guess that the pSB1C3_001_CMV did NOT work since the control with BLA looks nearly the same. We should repeat the experiment by cloning the CMV promoter into another vector. I would recommend it because we have a lot of constructs which we should fuse to the CMV promoter and in order to avoid the three-fragment ligation.
Transformation was performed according to the standard protocol w BL21 cells.
Comment: All N-terminal fusion approaches with VP2/3_HSPG-KO revealed positive results except of pCerulean_Zegfr:1907_MiddleLinker_VP2/3_HSPG-KO. Another clone was picked, preped, test digested and sent for sequencing.
Conclusion: Sequencing results revealed positive results.
Unfortunately there grew a bacteria lawn over night - it was hardly not possible to pick clones. Nevertheless I tried and picked 2 clones of pGA14_MiddleLinker_VP2/3_insCap and pGA14_MiddleLinker_VP2/3_HSPG-KO.
To do: Mini-Prep and test digestion.
Comment: For the creation of our super constructs, the His-Tag and the BAP motif need to be cloned into VP2/3 for N-terminal fusion to VP2. Sequencing results showed that the 6xHis and BAP motif was not cloned into VP2/3_insCap.
To do: Clone 587-KO_6xHis and 587-KO_BAP into pSB1C3_001_VP2/3_insCap 1. via digestion of inserts and 2. via hybridization of referring oligos - digestion of vector with 800-900 ng.
Conclusion: The VP2/3 fusion particle was transcribed and translated AND was transported back into the nucleus in order to be packaged by the ITR-flanked gene of interest.
Comment: Since the first gel revealed problems with samples 672, 673 and 682 another digestion was prepared (same approach as shown above), this time using a 0,8% gel and another loading dye (without SDS).
Comment: Since sample 672 showed no bands this sample was discarded and cloning continued only using 673.
Was performed according to protocol.
Ligation was performed over night.
Will be done tomorrow using BL21 cells.
New BL21 cells ready to use!
Comment: Unfortunately, the last experiment revealed that subcloning of the two ViralBricks did not work out which was shown by the sequencing results. Therefore we need to repeat the approach of subcloning the ViralBricks into the pSB1C3_VP2/3 which than can be used for fusing it to the n-terminal targeting approaches. Two different strategies will be carried out which means that we are digesting the pSB1C3_Viralbricks AND hybridize the oligos (BAP and His) in order to obtain some positive results.
The ligation was performed over night at 18°C with T4-ligase.
Comment: We decided to choose some additional GOI´s for the vectorplasmid so the "new" GOI´s need to be fused to the leftITR_Promoter_betaglobin constructs. Since only one clone grew on the plate prepared by Kira where she fused the CD (cytosine deaminase) to the above mentioned construct I am going to repeat it aswell.
Comment: In order to fuse the DARPin to the N-terminus of VP2/3_insCap and VP2/3_HSPG-KO, these motifs need to be fused to the Middle Linker (performed on 30.9.2010). In order to be able to immediately continue with cloning, BL21 cells were transformed and clones were picked in the noon. Now, the DNA needs to be preped and test digested in order to perform a 3-fragment-ligation with the DARPin and the pSB1C3_001_CMV plasmid. Unfortunately the cells of the VP2/3_insCap construct grew not densely enough. Therefore two new clones were picked from the plate and were inoculated in 10 mL LB medium. The other construct (pGa14_MiddleLinker_VP2/3_HSPG-KO) was preped and test digested.
Agarose gel: 0.8 %, 115 Volt, 1 hour.
Mini-Prep and test digestion of pGa14_MiddleLinker_VP2/3_ins Cap.
3 fragment ligation of MiddleLinker_VP2/3_ins Cap or MiddleLinker_VP2/3_HSPG-KO + DARPin + pSB1C3_001_CMV backbone.
Comment: Sequencing results show that all plasmids contain the HSGP-ko. Working will be continued using P644, P646 and P687.
You can either find these constructs in my workingfolder "DARPin" or the pGA14_MiddleLinker in the "Linker-Folder" and the VP2/3_insCap in many workingfolders!!!! You could have also digest the construct with EcoRI and SpeI... then you would expect a 2 and a 2.3 kb fragment... menas: you have to run the gel at least 1 hour - but it works (see the previous test digestion of the MiddleLinker_VP2/3_insCap construct!!!!).
Digestion with EcoRI & SalI; should yield two bands of 3400 & 1000 bp. The negative control should only be cut once.
Both samples show correctly sized bands. The negative control ran further than expected, it should be located at 4500 bp.
There are 45 samples to FACS on monday 4.10. Tripple values for each stock of each cell lines.
Comment: Ligation was carried out over night and transformation will be performed today.
9 6-well plates were prepared for the transfection tomorrow. 480000 cells were seeded into each well.
Comment: We also want to test the DARPin_E01 for the N-terminal fusion approaches. Today a 3 fragment ligation will be performed in order to complete the VP2 fusion constructs with the DARPin.
Because 2 test digestions showed that the fusion of the DARPin to VP1up_NLS didn't work as ecpected (insert is too large) this has to be repeated before also the VP1 insertion with the DARPin can be completed.
Was performed following the standard protocol.
Comment: Since we are still waiting for the ordered TK30, idea was to mutate the PstI site in the TK30 region. After mutating the PstI it is ready for submitting. Addtionally the ordered and received sr3ß can be subcloned.
After the PCR program was conducted, I added 1 µL of DpnI provided with the Kit to each sample and incubated it for 10 minutes.
For transformation XL-10 Gold cells with 2µL beta-mercaptoethanol have been used and plated on agar plates containing chloramphenicol.
Something happened to our virus-production cell line AAV293.
Nearly all AAV293 cells are dead.
The whole Loop-Insertion Transfection from 3.10 is lost, our actual culture flasks aswell. We dont know exactly the cause of this cellular mass mortality. Maybe someting is wrong with the new FCS-stocks.
The Test Digestion (1.10.) of the last attempts clones didn't look convincing. We sent clone 1 for sequencing yesterday, but we didn't recieve any results yet. That's why I decided to repeat the experiment today, in case results are negative.
Details: See first run from 28.9.
The two new standard vectors are from now on called Control Vectors, to get valide data. The TKGMK Stock will be used for the MTT-ASSAY.
Comment: Second approach of Cloning lITR_phTERT_beta-globin_mGMK_TK30 into pSB1C3_hGH_rITR, first one didn't work (see labday 01.10.).
Was performed according to the standard protocol.
Was done following the standard protocol using BL21 cells.
Was performed according to standard protocol using BL21 cells.
Results:The expected bands should run at 2035 and 2460 bp. The experiment has been repeated already today.
Comment: Cloning of 587-ko_His and 587-ko_BAP into pSB1C3_001_VP2/3_capins did not work out last time. Since cloning of VP2/3_capins started before sequencing results arrived, this experiment has to be repeated.
Comment: this approach was already repeated at least 2 times, but still even if its not one of the very important constructs, it should be finished by end of the month.
The gel shows only the primer bands on the bottom of the gel --> PCR failed again. Its going to be repeated tomorrow.
Not only the 293 cells look bad. The A431 cells and the HT1080 cells die, too. There is no contamination detectable. Maybe there is something wrong with the DMEM because the now 100 % serum-free AAV293 cells dont die.
The cells are not adherent so no transduction was performed.
Comment: After test digestion revealed positive results, we sent one clone for sequencing (P690) with VF-2 and VR-2 primers. The tube numbers are: SB_1 and SB_2.
Prefix looks well. Alle restriction sites can be found in the prefix RFC25.
Suffixlooks well. Alle restriction sites can be found in the suffix RFC25.
BUT: There seems to be a PstI recognition site within the sequence. Therefore the site-directed mutagenesis needs to be repeated.
Comment: Since the sequence analysis revealed that the PstI restriction iste within the CD is NOT removed I prepared another Quickchange in order to mutate the PstI site.
After the PCR program was conducted, I added 1 µL of DpnI provided with the Kit to each sample and incubated it for 10 minutes at 37°C.
Sequencing reults of pSB1C3_CMV_ZEGFR:1907_VP2/3_CapIns (P679), pSB1C3_CMV_ZEGFR:1907_VP2/3_CapIns_HSPG-ko (P684), pSB1C3_001_VP2/3_587_HSPG-ko_His(P734) and pSB1C3_001_VP2/3_587_HSPG-ko_BAP(P738). In P679 and P684, CMV_ZEGFR:1907 was cloned in front of VP2/3, in P734 and P738, 587_HSPG-ko_His/BAP was cloned into the 587 loop.
Comment: Besides the Affibody we also want to test and characterize the DARPin E01 as a targeting motif for EGFR over expressing tumor cells. The day before yesterday a 3 fragment ligation was performed with the CMV promoter, DARPin and pSB1C3_VP2/3_insCap/pSB1C3_VP2/3_HSPG-KO.
2 clones were picked from each approach and preped today.
Conclusion: Test digestion looks well: Expected fragments = 756 bp and 4409 bp.
Two samples were sent for sequencing to GATC using the CMV-F primer.
The biotinylation of the A20 antibody that recognizes the assembled virus was performed with a biotinylation Kit purchased from Dojindo.
The first biotinylation reaction was performed according to the protocol with 150µg of the purified antibody but resulted in not detectable protein concentrations.
The biotinylation reaction was repeated for a second time with 200µg and resulted in satisfying protein concentrations.
Successful biotinylation has to be tested in an other experiment.
Regarding the primer sequences, DMSO will be added to the PCR samples.
Comment: In order to perfom PCR, we have to clone the ordered Rep gene into Rep52 and Rep78 backbones.
Regarding the activity conditions of the enzymes, digestion was performed in 2 steps.
the gel shows that Rep78 backbone got lost.. Regarding the concentration, which was measured after PCR purification it might be possible --> digestion of Rep78 will be repeated tomorrow, while Rep52 is ready for ligation.
COMMENT: The DARPin was fused with the Middle Linker to the VP2/3 BioBrick and cloned into the pSB1C3 backbone. Yesterday's test digestion looked well, so one clone of the VP2/3_insCap and one clone of the VP2/3_HSPG-KO construct was sent for sequencing.
CONCLUSION: Sequencing looked well: 3 fragment ligation was successful - because the CMV-F primer was used for sequencing, the CMV promoter was successfully inserted. As shown in the alignments also the DARPin was inserted. This is the first time that the DARPin was sequenced - thus the original sequence must also be OK!
These two constructs are finished and are ready to be tested in cell culture via qPCR, ELISA,... and can be stored in our BioBrick Box for submitting.
The same procedure but with 7 ul Rep78 (c=556 ng/ul). After the first digestion, dephosporylation and PCR purification were performed and DNA was measured.
As a control, the ordered Rep was used again.
the second digestion as performed despite the low concentration.
The gel shows once again no detectable DNA of Rep78.
The transformation was performed according to the standard protocol with BL21. Compared to VP1-3 transformed cells, Rep52 cells revealed no visible pellet after centrifugation, thus 300 ul of the cells suspension was plated.
Gel1_Part1: The 453 constructs were cut with BamHI and SspI, the 587 constructs with SalI and PvuII. The inserts were cloned into pSB1C3_RepCap_p5tatless.
Comment:Additionally, P771 and P781 from yesterday's Mini-Prep were digested again.
Comment:Samples P791 to P796 perform like expected, they can be used for further cloning or testing in cell culture. P771 show strange bands, a test PCR will be performed to verify successful cloning. P781 does not show any DNA at all like last time. A new SDm will be performed for pSB1C3_mGMK_TK30 (P675).
Comment: Test digestion showed strange results (an additional band at 500bp which does not fit any expectations), therfore an additional test-PCR was performed to verify successful cloning of the complete constructs.
Comment: Test PCR showed successful cloning of lITR_phTERT_beta-globin_mGMK_TK30 into pSB1C3_hgH_rITR. SDM to remove PstI in TK30 can be performed.
Comment: The first try to remove the PstI restriction site delivered no plasmid DNA, so another approach needs to be started.
To digest methylated and hemi-methylated DNA, 1 µl of DpnI was added and the mixture was incubated for 10 minutes at 37 °C.
Since XL-10 Gold cells delivered a bacteria lawn on the plates, this time XL1b cells were used.
Comment: Mini-Prep was performed according to standard protocol.
Comment: Since Mini-Prep from XL-10 cells yielded that little plasmid, a re-trafo will be performed using BL21 cells.
A 1:10 dilution of both plasmids was prepared and 1 µl was added to BL21 cells.
Transformation was performed according to standard protocol. Cells were plated on agar plates containing chlorampenicol.
Preparation of BL21 (red line) and XL1-blue (green X) cells was performed according to standard protocol.
Do not use cells until further notice!
Comment: The GOI which is flanked by the ITRs and encapsidated in the capsid is the mGMK_T30 driven by the pHTERT promoter which is only active in several tumor cells. Since we have never tested the primers if they work I wanted to do that. This time I tried to use the TAQ polymerase which we received from Peqlab. Since I could not find a standard protocol for the PCR provided by peqlab I tried to compare several manuals and combine them.
Protocol: I did not tried the approach with ENhancer Solution.
I checked the protocol today (10.10) and recognized that I added accidentially to much buffer to the PCR-Mix. Maybe this could be another explanation for the negative results of the PCR.
Unfortunately the expected PCR product with a size of 182 bp could not be detected as it can be seen in the agarose gel. I can not say if the protocol needs to be modified or if the primer pair did not anneal adequatly. Therefore, I am going to repeat the PCR tomorrow by using the Phsuion polymerase for checking the results.
For transformation BL21 cells have been used and plated on agar plates containing chloramphenicol.
Comment: Since we still have the PstI restriction site within the sequence we need to delete the restriction enzyme site.
For transformation XL-1 cells have been used because the bacterias of the last site-directed mutageneis approaches using XL-10 cells showes extensive grwoth (bacterial lawn). I plated on the transformaned cells on agar plates containing chloramphenicol.
In order to obtain transduction data for our targeting approaches, we still need constructs with RGD/Z34C in the 453 loop as well as HSPG_KO_empty in the 587 loop.
I digested pSB1C3_HSPG-KO_empty (P611) as well as pSB1C3-001_pCMV_[AAV2]VP123_453_RGD (P634) and pSB1C3-001_pCMV_[AAV2]VP123_453_Z34C (P641)with BamHI and PvuII and ligated the HSPG-KO_empty motive into both vectors.
the gel picture for the insert has not been saved. But it did show a faint band at ~50 bp.
mVenus without beta-globin: beta globin is said to improve the expression therefore we expect a worse mVenus expression compared to the standard AAVs.
mVenus without HGH: HGH is responsible for the mRNA polyadenylation of the virus so we expect expect a worse mVenus expression compared to the standard AAVs, too.
mVenus HSPG KO: the primary AAV2 receptor (the HSPG receptor) was knocked out therefore we expect a worse tranduction rate (about 0,3 to 0,4 of the standard transduction rate, assuming that there are HSPGs on the HT1080 and A431 cell surface). The worse the better.
mVenus new standard: AAV2 with 4 point mutations, inserted rep & cap and "remutated" KpnI restriction site hopefully yiels results similar to the mVenus standard.
Comment: The GOI which is flanked by the ITRs and encapsidated in the capsid is the mGMK_T30 driven by the pHTERT promoter which is only active in several tumor cells. Since we have never tested the primers if they work I wanted to do that. For several reason I decided to use the Phusion polymerase to perform the PCR. First, we are familiar with the handling of this polymerase and have a protocol which we already used several times and second, since I could not find any standard protocols for the TAQ-polymerase provided by PeqLab, as mentioned yesterday, I will rely on the Phusion polymerase.
Results: After PCR was completed, I loaded a 1.5% agarose gel. The results can be seen in the picture: There is a intensive band aat around 150-180bp which correspond to the expected fragment size of 182 bp. We can conclude that the the primers work and that they can be used in the qPCR approaches. One note along the way: I´ve missed the fact that the buffer in which the gel was loaded, was TBE and not TAE. Still the fragment looks well, even though the gel did not appeared to ran normal.
New XL1b and BL21 cells ready to use!
Comment: In order to send our final VP2 N-terminal Fusion constructs with HSPG konock down and our super constructs to the registry, they need to be cloned into the pSB1C3 backbone.
We would prefer sending them in the pSB1C3_001 plasmid, because it does its ViralBricks restriction sites were deleted. Therefore one can cut out the 587_KO ViralBrick and replace it by any ViralBrick of interest.
Because test digestion of pSB1C3_001_CMV didn't show any clear results, I will digest both clones and in addition to that pSB1C3_CMV.
Comment: Unfortunately, no cells grew on the agar plates containing cm. Therefore, we repeat the Trafo ones again and plate them on new agar plates hoping that colonies will grow over night.
Two 96-well plates that were coated with 200µg A20 antibody per well for 48h at 4°C were washed three times with 200µl PBST Buffer and incubated over night at 4°C with 300µl PBST + 0.2 % BSA.
As a preparation 4 liters of PBS buffer were prepared.
Comment: Vector was cut using PstI in order to verify successful deletion of the PstI restriction site in TK30. As it can be seen above, both vectors are linearized, no fragment was cut out. In case of failed SDM, one additional band at roughly 450 bp was expected. Since this band does not show up, successful deletion is confirmed.
Comment: In order to test our GOI with another promotor, lITR_phTERT_beta-globin_mGMK_TK30 needs to be cloned upstream of hgH_rITR.
Since ligation with Quickligase (because of one blunt end) revealed no colonies on the plate, I decided to perform ligation with T4 ligase.
Comment: Because yesterday's cloning didn't deliver a good separartion of the expected gel bands. Nevertheless ligation and trafo was performed. In order to immediately find out, whether we received successful results a colony PCR will be performed.
Two clones were picked from each plat. In addition to that a positive (pAAV_RC) and a negative control (pSB1C3_lITR) was prepared.
Used primer: 4200 rev and Cap3500 for. Expected fragment size: 885 bp.
PCR was performed following the standard protocol.
All samples match the positive control!
To do: Mini-Prep and sequencing.
Comment: In order to perform a Western Blot of different virus capsids (with and without capsid-motifs), 2 10% SDS polyacrylamid gels were prepared.
Comment: Since the first attempt did not work,and no cells grew on the plate and the same ligation was transformed again into BL-21 and a lot of clones grew on the plate, I decided to perform a colony PCR in order to check several colonies and to inoculate at the same day for a Midi-Prep.
Result: We can see two things: The cloning of p40 to VP123 worked quiet well AND the Robust PCR Kit which was used for the first time worked as well.
Comment: To produce another GOI for testing in cell culture, the cytosine deaminase needs to be assembled with lITR_promotor_betaglobin. In the next step hgH_rITR needs to be added.
CD clone 1 yielded to bands around 2500 bp to 3000 bp. Since the vector was cut only using EcoRI and SpeI, it was expected to be linearized, not to be cut into two fragments this size. Therefore, this sample was discarded and cloning was continued using CD clone 2.
The AAV particle standard that contains 3.6x10^9 viral particles was dissolved in 500µl as described in the protocol of the Progen AAV Titration ELISA and a absorption spectrum was measured. These purified and concentrated viral particles could be used for biophysical measurements, there for the possibility to detect the viral particles by absorption was interesting for us.
T4 ligation seems to worked out. The plate contained many colonies.
Comment: Sequencing results revealed that there is CFP rather than CD in P819. In order to verify this and exclude the possibility that the wrong plasmid was sent for sequencing test digestions were performed. Additionally the original vector at which the SDM to remove the PstI was performed was digested as well to verify its insert.
Comment: Using these digestion approaches, it was expected for CFP to deliver always bands at ~750 bp. For the CD it was expected to show bands at 90 bp, 1200 bp and 2000 bp if the CD still contains the PstI restictiton site, 1300 bp and 2000 bp if it was removed.
Comment: As it can be seen, P819 does contain CFP, therefore a mistakenly sequenced plasmid can be excluded. Also it can be seen that P690 contains the CD. Therefore an additonal SDM of P690 to remove the PstI restriction site is necessary.
Virus Stock: A.F.; P816; TKGMK-Standard; R/C: P468; 5.10; consists of supernatant from pelleted cell fragments.
Following the Protocol used by (Rohr et al., 2002).
I digested 5 µl virus dilution with 7,5 µl DNAse I (7,5 units) and 25 µl 50 mM MgCl2 (end concentration 25 mM) in a final volume of 50 µl at 37°C for 30 min.
è DNAse should be heat inactivated at 65°C for 10 min. I forgot that step. The enzyme should however be inactivated in the initial denaturation step.
è Both negative controls containing primers were contaminated with DNA.
è Negative controls containing only H2O were negative.
è Because of the contaminated negative controls and the high deviation, I will repeat the experiment tomorrow to verify the results.
Rohr, U., Wulf, M., Stahn, S., Steidl, U., Haas, R., Kronenwett, R., et al. (2002). Fast and reliable titration of recombinant adeno-associated virus type-2 using quantitative real-time PCR. Journal of virological methods, 106(1), 81-8. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/12367732.
Comment: We do not only want to submit the tk30, a mutant version of the thymidine kinase, but as well another mutant protein which is named sr39. SR39 seems to have a better activity than the tk30 without being fused to the guanylate kinase (mgmk), but the sr39 also works quite well as a fusion protein. We ordered one part of the sr39 and we are now subcloning it into the mgmnk_tk30 construct in order to obtain mgmk_sr39.
The digested fragments were loaded on a 1.2% agarose gel. The results can be seen above in the gel picture: I cut out the small visible band in the right lane, and the upper intensive band of the left lane.
After gel extraction has been performed, I ligated the two fragments and transformed BL-21 cells, plated them on cm agar plates and incubated them over night at 37°C.
For the first Western Blot try, we investigate whether is is enough to concentrate the virus stock (from cell supernatant) and then to simply load it onto a SDS-PAGE gel and perfom immuno-staining.
For this purpose 8 mL virus stock (pHelper, pAAV_RC, GOI=YFP) were concentrated via amicon filtration.
First, amicon filter was washed with 4 mL PBS.
Then 4 mL virus stock was loaded onto the filter and centrifuged at 3000 rpm for 15 minutes.
Flow-through was discarded and additional virus stock was loaded, centrifuged at 3000 rpm for 25 minutes.
Flow-through was discarded, remaining solution was re-suspended and additional virus stock was added. Amicon filter was centrifuged at 3000 rpm for 50 minutes.
Flow-through was discarded. Residual protein solution: 500 µL. Filter was washed with 3 mL PBS, centrifuging 35 minutes two times.
2 x 40 µL protein solution (16x) was taken and mixed with Laemmli buffer and stored over night @ 4°C.
To do: SDS-PAGE and Western Blot tomorrow.
Perfomance of another try by harvesting viruses of the cell pellet with RIPA buffer & freeze/ thaw.
Western Blot of unmodified, and modified capsids (N-terminal fusion and VP1 insertion with CFP ref. mVenus) --> size shift should be detectable.
Comment: P822 and P823 smear a lot, therefore evaluation is difficult. Another test digestion would be reasonable.
P824 contains hgH_rITR, therefore it will be discarded. P825 looks well and was sent for sequencing.
P830 and P831 look well and will be sent for sequencing tomorrow.
Comment: Sequencing revealed that there is no CD in our pSB1C3_CD but rather CFP. So, anonther approach of the SDM needs to be performed.
Trafo was performed according to protocol, but using XL1b cells instead of XL-10 Gold cells.
Comment: Yesterday a 8x concentration of AAV2 supernatant-stock was performed. 10 µL Laemmli-buffer was added to 40 µL AAV stock (8x) and stored over night at 4°C.
Today a SDS-PAGE of these samples will be performed.
Samples were incubated for 10 minutes at 95°C for denaturation.
After running front reached the end of the gel, PAGE was stopped.
Three layers of transfer buffer soaked whatman paper were placed onto the blotter. A polyvinylidene difluoride membrane was activated by methanol (5 seconds) and then incubated for 5 minutes in transfer buffer. Membrane was then placed onto the whatman paper, gel was carefully layed on top. Air bubbles were removed, ensuring elictricity flow. After coverage by 3 further layers of whatman, blotting was performed at 200 mA for 1 hour.
In order to minimize nonspecific antibody interactions, membrane was incubated in blocking buffer (TBS (pH 7.5), 5% milk powder) for one hour, shaking.
A 1:10 dilution of the B1 antibody in TBS + 1% milk was prepared. Membrane was incubated in 50 mL falcon over night at 4°C, rotating.
Comment: Repetiton of yesterday's test digestion which yielded no clear band at 2000 bp as it was expected.
Comment: This time both samples look well. P822 will be sent for sequencing.
Sequencing comfirmed that the pysical sequence fits our theoretical. Next step will be to remove the remaining PstI restriction site within TK30.
Dephosphorylation was performed because of the blunt cleavage. PCR phosphorylation followed in order to get rid off the buffer because Buffer 3 is not appropriate for further digestion.
Comment: Yesterday, SDS PAGE, Western Blot and primary B1 mouse antibody incubation was performed.
Today, secondary antibody incubation and ECL analysis will be done.
Membrane was washed 3 x 7 minutes in TBST (0.2% Tween).
A 1:5000 dilution of secondary goat anti-mouse HRP (sc2005) coupled antibody in TBS was prepared.
Membrane was incubated in secondary antiobody for 1 hour.
Membrane was washed 3 x 10 minutes.
Analysis of the membrane revealed that the samples didn't run well. Much unspecific antibody binding was detectable.
Because of these dissatisfying results SDS PAGE will be repeated. Instead of performing Western Blot, gel wioll be stained by Coomassie in order to optimize SDS-PAGE conditions.
As usual the PSB1C3 bands indicate a higher bp amount than they actually got.
... and following ligation (2 µl Insert, 6 µl vector, 1 µl T4 DNA Ligase, 1 µl T4 DNA Ligase Buffer, 40 minutes, RT) were performed according to the standard protocol. After the transformation (with BL21) the cells were plated and put into the 37°C room.
These ViralBricks were not ordered as hybridized oligos, but can be cloned from the Cap-Gene-Synthesis into the ViralBrick MCS.
Comment: In order to optimize SDS PAGE with the viral capsid proteins, I didn't perform Western Blot, but Coomassie staining.
Two different virus stocks with unmodified capsid were taken. 2 samples were prepared of each: One sample was untreated, the other one was centrifuged in order to sediment cell fragments.
10 µL Laemmli buffer (5x) was added to 40 µL virus sample and incubated at 95°C for 5 minutes.
Gel was run 1.5 hours at 10 mA.
Gel was put into coomassie bath, heated in microwave and incubated for 1 hour, shaking.
In order to remove background coomassie staining, gel was incubated in acetic acid (20%) for 2 hours, shaking.
Results revealed that samples ran well (vertically :) ) - 20 µL seems to be too much. Interestingly centrifuging the virus stocks in order to remove cell debris didn't lead to better results.
To do: Perform Western Blot again - with 10 µL Virus stock, 0.5% instead of just 0.2% TBS-milk and longer washing steps.
Comment: We assembled targeting super constructs which specifically bind the EGF receptor with VP2 N-terminal fused ZEGFR:1907 Affibody and can either be purified via a loop inserted His-Tag or visualized via Streptavidin coupled fluorescent proteins.
Further on these loop motifs were vloned to the "mVenus VP1 insertion".
Sequencing results showed that insertion of these constructs into pSB1C3_CMV worked!
Next steps: Put sample sinto BioBrick Box.
Comment: Transfection was performed in order to evaluate the promoter activities.
Yesterday, the cells were seeded in the 6 well plate by Sven. Transformation was performed according to the manufacturer protocol (Qiagen).
Transformation was successful. 2 clones will be picked for mini prep tomorrow.
Rep78 and Rep52 were picked (2 clones from each plate), in order to continue with mini prep and PCR amplification.


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text:
Mario Novelli, né à Rome le  et mort dans la même ville le , est un acteur italien parfois crédité sous le nom de Anthony Freeman.

Biographie

Il est le père des acteurs Alessandro et Emiliano Novelli.

Filmographie partielle
Acteur
 1963 : Ursus gladiatore ribelle, réalisation de Domenico Paolella
 1964 :
Gli schiavi più forti del mondo, réalisation de Michele Lupo
La vengeance de Spartacus  (), réalisation de Michele Lupo
Gli invincibili fratelli Maciste, réalisation de Roberto Mauri
1965 :
I tre centurioni, réalisation de Roberto Mauri
Sette contro tutti, réalisation de Michele Lupo
Per una manciata d'oro, réalisation de Carlo Veo
1966 :
Texas Adios (), réalisation de Ferdinando Baldi
Destination: planète Hydra (), réalisation de Pietro Francisci
1967 :
Deux croix pour un implacable () (), réalisation de Rafael Romero Marchent
Ballade pour un pistolero (), réalisation de  Alfio Caltabiano
Un homme, un cheval et un pistolet (), réalisation de Luigi Vanzi
Trois supermen à Tokyo, réalisation de Bitto Albertini
1969 :
Fräulein Doktor, réalisation d'Alberto Lattuada
Isabella duchessa dei diavoli, réalisation de Bruno Corbucci
1970 :  La Nuit des assassins (), réalisation de Adriano Bolzoni
1971 :
Abattez Django le premier (), réalisation de Sergio Garrone
Ma dernière balle sera pour toi! () (), réalisation de Aldo Florio
Warnung vor einer heiligen Nutte (), réalisation de Rainer Werner Fassbinder
Ore di terrore, réalisation de Guido Leoni
Acquasanta Joe, réalisation de Mario Gariazzo
1972 :
Milan calibre 9 (), réalisation de Fernando Di Leo
Un uomo chiamato Dakota, réalisation de Mario Sabatini
1973 :  Piège pour un tueur (), réalisation de Massimo Dallamano
1975 :
Rome violente (), réalisation de Marino Girolami
Bracelets de sang (), réalisation de Umberto Lenzi
Faccia di spia, réalisation de Giuseppe Ferrara
L'albero di Guernica, réalisation de Fernando Arrabal
1976 :
Come cani arrabbiati, réalisation de Mario Imperoli
Le Désert des Tartares (), réalisation de Valerio Zurlini
I padroni della città, réalisation de Fernando Di Leo
Liebes Lager, réalisation de Lorenzo Gicca Palli
Paura in città, réalisation de Giuseppe Rosati
La lozana andalusa (La lozana andaluza), réalisation de Vicente Escrivá
Hanno ucciso un altro bandito, réalisation de Guglielmo Garroni
1977 : Adios California, () réalisation de Michele Lupo
1978 :
Coup de gueule (), réalisation de Gianni Martucci
La Quatrième Rencontre (), réalisation de Mario Gariazzo
Scorticateli vivi, réalisation de Mario Siciliano
Le Crime du siècle  (Indagine su un delitto perfetto), réalisation d'Aaron Leviathan
1979 : Play Motel, réalisation de Mario Gariazzo
1980 : Les novices libertines (), réalisation de Bruno Mattei
1982 :
Les aventures sexuelles de Néron et de Poppée (), réalisation de Bruno Mattei
Le notti segrete di Lucrezia Borgia, réalisation de Roberto Bianchi Montero
Crime au cimetière étrusque (), réalisation de Sergio Martino
Sangraal, la spada di fuoco, réalisation de Michele Massimo Tarantini
Vai avanti tu che mi vien da ridere, réalisation de Giorgio Capitani
1984 : 2072, les mercenaires du futur (), réalisation de Lucio Fulci
1987 :  Delta Force Commando, réalisation de Pierluigi Ciriaci
1989 :
Afghanistan - The last war bus  (), réalisation de Pierluigi Ciriaci
Evil Train ( (Beyond the Door III), réalisation de Jeff Kwitny
1991 :  Year of the Gun, l'année de plomb () (Year of the Gun), réalisation de John Frankenheimer
1998 :  Coppia omicida, réalisation de Claudio Fragasso
2000 :  Alex l'ariete, réalisation de Damiano Damiani

Notes et références

Liens externes
.

Naissance à Rome
Naissance en février 1940
Décès à Rome
Acteur italien
Décès en août 2016
Décès à 76 ans

redpajama_set_name: RedPajamaWikipedia


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A nice piece of history about this cocoa plantation can be found on the website of Pump Street. The only Jamaican estate that grows, ferments and dries high-quality cocoa itself.
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text:
Act I, scenes i–iii
Act I, scenes iv–v; Act II, scenes i–ii
Act II, scenes iii–vi
Act II, scene vii; Act III, scenes i–iii
Act III, scenes iv–vii
Act III, scenes viii–xiii
Act IV, scenes i–viii
Act IV, scenes ix–xv
Act IV, scene xvi–Act V, scene i
Act V, scene ii
Mark Antony
Octavius Caesar
How To Cite No Fear Antony and Cleopatra
Themes are the fundamental and often universal ideas explored in a literary work.
The Struggle Between Reason and Emotion
In his opening lines to Demetrius, Philo complains that Antony has abandoned the military endeavors on which his reputation is based for Cleopatra's sake. His criticism of Antony's "dotage," or stupidity, introduces a tension between reason and emotion that runs throughout the play (I.i.1). Antony and Cleopatra's first exchange heightens this tension, as they argue whether their love can be put into words and understood or whether it exceeds such faculties and boundaries of reason. If, according to Roman consensus, Antony is the military hero and disciplined statesmen that Caesar and others believe him to be, then he seems to have happily abandoned his reason in order to pursue his passion. He declares: "Let Rome in Tiber melt, and the wide arch / Of the ranged empire fall" (I.i.35–36). The play, however, is more concerned with the battle between reason and emotion than the triumph of one over the other, and this battle is waged most forcefully in the character of Antony. More than any other character in the play, Antony vacillates between Western and Eastern sensibilities, feeling pulled by both his duty to the empire and his desire for pleasure, his want of military glory and his passion for Cleopatra. Soon after his nonchalant dismissal of Caesar's messenger, the empire, and his duty to it, he chastises himself for his neglect and commits to return to Rome, lest he "lose [him]self in dotage" (I.ii.106).
As the play progresses, Antony continues to inhabit conflicting identities that play out the struggle between reason and emotion. At one moment, he is the vengeful war hero whom Caesar praises and fears. Soon thereafter, he sacrifices his military position by unwisely allowing Cleopatra to determine his course of action. As his Roman allies—even the ever-faithful Enobarbus—abandon him, Antony feels that he has, indeed, lost himself in dotage, and he determines to rescue his noble identity by taking his own life. At first, this course of action may appear to be a triumph of reason over passion, of -Western sensibilities over Eastern ones, but the play is not that simple. Although Antony dies believing himself a man of honor, discipline, and reason, our understanding of him is not nearly as straight-forward. In order to come to terms with Antony's character, we must analyze the aspects of his identity that he ignores. He is, in the end, a man ruled by passion as much as by reason. Likewise, the play offers us a worldview in which one sensibility cannot easily dominate another. Reason cannot ever fully conquer the passions, nor can passion wholly undo reason.
The Clash of East and West
Although Antony and Cleopatra details the conflict between Rome and Egypt, giving us an idea of the Elizabethan perceptions of the difference between Western and Eastern cultures, it does not make a definitive statement about which culture ultimately triumphs. In the play, the Western and Eastern poles of the world are characterized by those who inhabit them: Caesar, for instance, embodies the stoic duty of the West, while Cleopatra, in all her theatrical grandeur, represents the free-flowing passions of the East. Caesar's concerns throughout the play are certainly imperial: he means to invade foreign lands in order to invest them with traditions and sensibilities of his own. But the play resists siding with this imperialist impulse. Shakespeare, in other words, does not align the play's sympathies with the West; Antony and Cleopatra can hardly be read as propaganda for Western domination. On the contrary, the Roman understanding of Cleopatra and her kingdom seems exceedingly superficial. To Caesar, the queen of Egypt is little more than a whore with a flair for drama. His perspective allows little room for the real power of Cleopatra's sexuality—she can, after all, persuade the most decorated of generals to follow her into ignoble retreat. Similarly, it allows little room for the indomitable strength of her will, which she demonstrates so forcefully at the end of the play as she refuses to allow herself to be turned into a "Egyptian puppet" for the entertainment of the Roman masses (V.ii.204).
In Antony and Cleopatra, West meets East, but it does not, regardless of Caesar's triumph over the land of Egypt, conquer it. Cleopatra's suicide suggests that something of the East's spirit, the freedoms and passions that are not represented in the play's conception of the West, cannot be subsumed by Caesar's victory. The play suggests that the East will live on as a visible and unconquerable counterpoint to the West, bound as inseparably and eternally as Antony and Cleopatra are in their tomb.
The Definition of Honor
Throughout the play, characters define honor variously, and often in ways that are not intuitive. As Antony prepares to meet Caesar in battle, he determines that he "will live / Or bathe [his] dying honour in the blood / Shall make it live again" (IV.ii.5–7). Here, he explicitly links the notion of honor to to that of death, suggesting the latter as a surefire means of achieving the former. The play bears out this assertion, since, although Antony and Cleopatra kill themselves for different reasons, they both imagine that the act invests them with honor. In death, Antony returns to his identity as a true, noble Roman, becoming "a Roman by a Roman / Valiantly vanquished" (IV.xvi.59–60), while Cleopatra resolves to "bury him, and then what's brave, what's noble, / Let's do it after the high Roman fashion" (IV.xvi.89–90). At first, the queen's words seem to suggest that honor is a distinctly Roman attribute, but Cleopatra's death, which is her means of ensuring that she remains her truest, most uncompromised self, is distinctly against Rome. In Antony and Cleopatra, honor seems less a function of Western or Eastern culture than of the characters' determination to define themselves on their own terms. Both Antony and Cleopatra secure honorable deaths by refusing to compromise their identities.
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Antony and Cleopatra: Popular pages
NO FEAR No Fear Antony and Cleopatra
CHARACTERS Mark Antony: Character Analysis


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Board Hires ISG For Pipeline Inspection
The Mahaska County Board of Supervisors accepted a letter of intent for county representation and inspection services for the Heartland Greenway System project from ISG at the Nov. 15 regular meeting.
Representative Evan Del Val of ISG, a Des Moines-based engineering firm, spoke with the board about ISG's county inspection services.
The Heartland Greenway project not yet finalized, but is expected to begin in 2024.
It is an approximately 1,300-mile carbon capture and sequestration network that will provide a path towards carbon neutrality for some of the largest commercial producers of CO2 in the heartland by capturing CO2 emissions and transporting them into a secure underground site.
ISG's services would include assigning inspectors to construction crews to ensure quality completion of the project.
Inspectors will be assigned to focus on specific aspects of construction, including top soil stripping and trenching, among others.
Landowners with complaints about how the project is being completed will be able to reach out to ISG for guidance or assistance.
Board of Supervisors chair Mark Groenendyk reminded constituents that the board does not have a say on the details of the project.
"This board has no authority on whether the pipeline goes through, where it goes, that's strictly up to the legislation," he said. "The only responsibility we have is to hire an inspector to make sure that the work is done properly. [ISG is] kind of the intermediary between the pipeline and the landowner."
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Driving Public Sector Digital Transformation
Chris Pope
Ronald Reagan once said, "The nine most terrifying words in the English language are: I'm from the government and I'm here to help."
It is always amusing (if slightly controversial) to start public sector IT presentations off with this quote, but it sets the tone, gets the blood flowing and it helps convey a sense of how (we very much hope) that time has passed.
And governments are changing. All around the world we see public administration bodies being compelled to reinvent themselves, become more dynamic and embrace all of the cloud-driven, often mobile-first, technologies that will ultimately lead them towards a new point of digital transformation.
Governments have to do this, because governments have to now reshape to serve connected smart-city societies, where individuals expect intuitive consumer-level interactions with every public service that they come into contact with.
Governments can no longer afford to be seen as slow moving, bureaucratic behemoths slowed down by the weight of their own red tape.
A fundamental reinvention at the core
To become proactive, forward-looking bodies, governments realise that they must now reinvent themselves at the core. This requires a shift towards digitally transformative technologies that will elevate back office processes, operations processes and commercial processes into a new digital space.
That new digital space is perhaps unsurprisingly driven by cloud computing – and in 2018, with the UK G-Cloud Digital Marketplace now established, there should be no misgivings about the use of the cloud computing as a public data resource.
But government digital transformation isn't just about moving existing systems to the cloud computing model. It is also all about using a new digital fabric to innovate for the future. Not always the type of thing you expect the public sector to talk about, this is where new products and services are developed and brought online, new public-private channels are forged and new business models and cemented.
The UK government has digitally progressed exceptionally well in relation to some other contemporary first world economies. As we now look to the UK's digital government future and further afield, we can see new innovations, such as micro-transactions over social media channels like WhatsApp, being used to perform a certain number of public services.
When we look at the pressure that the commercial world is putting on user experience, government is now having to understand how fickle consumers are today in relation to the way they are using their phone provider, utilities suppliers and even services such as Uber. This is the transformative shift that government must now embrace. The use of public services has to be efficient, always-on, collaborative, intuitive and perhaps even fun.
ServiceNow is aware that citizens can now form an invaluable information channel for government to use to operate with and improve service delivery in public sector. This can all be channelled towards populating a service portal so that all services can be managed from a holistic, fully-informed viewpoint.
Four factors in every service
If we consider what a service (any service, governmental or otherwise) relates to and is composed of, we know that all services are actions designed to:
Get help.
Make a request.
Make a change.
Provide a user with knowledge and advice.
In real terms, these four elements manifest themselves in any number of things, like visitor building registration processes for example, which are typically forms-based applications. This is the type of process that typically takes up to much time and manual effort inside any given organisation – and these are the kinds of things we can build within the ServiceNow platform in as little as 10 minutes.
We can then take that information from our forms-based applications forward and start to apply anomaly detection models to work out who should be where at what time so that we can analyse what kinds of workflow patterns are playing out across the business.
At this level we now see a lot of peer benchmarking where ServiceNow customers are using anonymised data in specific industry verticals to see how fast, for example, a financial services organisation would be able to fix and outage or fulfil a service request. If we can start to bring these new types of actionable analytics to bear then we can start to make the following type of judgement:
I know how (a public ministry or agency) performs historically.
I know how it is performing currently due to service portal analytics.
I know how other similar operations across peer review benchmarks are performing.
I know where this departments need to get to operationally.
I know what types of strategic business action you need to take next.
A wide adoption curve
On the total all-industry path to digital transformation we see chemical firms alongside oil and gas organisations and utilities all at the slow end of the curve. At the other 'born in the cloud' end of the curve we see tourism, banking and media at the most innovative business operations.
Government can use the breadth of this viewpoint as a measure to take individual ministries, and the sub-agent divisions that compose them, to strategically plan and build service-centric workflows for the future.
The new era of government digital transformation will work with a new services taxonomy that underpins service experience, service delivery and service assurance – all benefiting from a higher-level fabric of service analytics.
ServiceNow exists to automate business processes faster, more effectively and more efficiently. As we now help build digital government programmes far elevated above the data silo architectures of the past, we will help build a better society, for all societies.


Join Ashley Kennedy for an in-depth discussion in this video The power of music: Emotion, subtext, information, and rhythm, part of Introduction to Music in Film and Video.
- For anyone who's ever heard a note of music, it's clear that it's a very powerful thing. And in this course, we're gonna be talking about a special type of music, film score. Film score has tremendous influence on the viewer and the amazing thing is it primarily happens at a subconscious level. We watch the film, we pay attention to plot, character, and conflict, we feel the rising action and experience the climax with the characters. We focus on all of that, what we don't often consciously focus on is the accompanying musical score.
This seeps its way into our subconscious in amazing ways. Now, music's inclusion in film is almost as old as the motion picture itself, but the reasons behind it have certainly evolved. In the late 1800s and early 1900s, music wasn't synced to the movie at all, it was just used as accompaniment to the onscreen action initially for reasons as simple as masking the sound of the projector in an otherwise silent theater. And because those early films lack dialogue, music initially helped audiences stay engaged by providing the appropriate audio backdrop.
But it wasn't long before music was used to enhance the mood of specific moments on screen and connect to audiences in meaningful ways. Early on, movie houses employed pianists and organists and even entire orchestras to play music during silent films. Music cue sheets and sometimes full scores were often distributed with the prints of the film so they could be played along. And when cue sheets weren't available, the musicians improvised to provide an appropriate emotional shape to each main character and plot point, syncing their musical cues to the film on the fly.
Since those days, the basic principles of using music to emotionally enhance the picture have remained in tact and have grown exponentially. There were actually brief movements in the 1950s and later in modern times when pockets of realist and purist filmmakers rebelled against music in film. But those movements certainly didn't last. Audiences demanded music, and even more, films themselves demanded it. So we're gonna talk all about music's role in film in this chapter and in this movie I'm gonna break things down on a high level.
First, music impacts emotion, a lot. In a basic sense, this is the same as music you would listen to on its own. Slow, soft music most often calms. Fast, vibrant music excites. There are different musical strategies to evoke tension, uneasiness, romance, bravery, sadness, and dozens of other ways of feeling. Now take all of that and put that with the moving image. When combined, music can put thoughts and feelings in the mind of the viewer to produce a totally new or an elevated result.
I wanna start very simple, not with a scene from a movie but just with a single shot. Take a look at this short clip of an aerial dancer descending down a silk. We don't see her face, we're just getting a general sense of movement. Now let's take a look at this same shot but with different musical undertones. (slow reed instrument music) (calming piano music) (suspenseful music) As you can see, music can make us feel different things about what we see even when what we see doesn't have much emotion attached to it to begin with.
But it's this juxtaposition of image and sound that is so often explored and sometimes exploited, reaching into our subconscious in ways that images alone simply can't. Now I wanna talk more about this notion on a deeper level because music doesn't just affect the way viewers feel about a particular shot or scene, it also offers clues about the subconscious of the characters within the film. It acts as a type of subtext. Because, think about it.
When you're reading a book or a script or some other text, you get plenty of clues in commentary about the character's own inner thoughts and emotions through the written word. The author often tells you what the characters are thinking or at least gives you enough to make your own deductions. It's because you get so many more layers of information in the written form that many prefer a good book to the movie's adaptation of it. Now you can certainly pick up on some of that in the form of great acting and cinematography, sound design, and other cinematic tools.
But it's the role of music that plays a large role in giving us this subtext. So let's take a look at another very simple shot. This time focusing on a person whose face you can see. It's just a guy drinking a beer. We don't know what he's thinking, and the video doesn't give us many clues. But now what do you think he's thinking? (sad music) Or now? (upbeat drum music) Or how about now? (pensive music) So again, now we're going beyond how we feel as the viewer and we're transposing different types of subtext on what this character could be thinking and feeling.
What else? Well music can also provide us information about things like setting, culture, nationality, and even time period. Every area of the world and most periods of time have specific music characteristics and filmmakers use that all the time. Take a look at this slow panning shot of some mountains and trees, which we'll assume is the opening establishing shot of a documentary. See if you can tell what clues you might get about this setting or culture. Perhaps it's in Asia. (Asian music) Or Africa.
(African drum music) Or an island nation. (tropical music) Truth be told, it's actually in California but as you can see, music is a tool that can help establish information and context even before we're provided with any actual details in the film. In fact, this is used so often that film score has actually contributed to many stereotypes in regions and cultures around the world, so just be aware of that.
Sometimes you're hearing what people think is the music of a place rather than the authentic music of a place. Next, I wanna talk about how music can set a rhythm to a scene that may not have inherently been there before. And of course this links back to emotion. There are many ways to think about this, and countless examples, but let me just talk about a couple of different musical strategies that can help evoke emotion through rhythm. For example, when composers want to convey calm and tranquility, they might use smooth, quiet musical elements.
They'll use notes of longer duration and they'll also use silence. In this scene from American Beauty, a single shot lasts for more than a minute. It's just one shot, along with a few thoughtful, sporadic lines of dialogue. It's the music though, that helps to mesmerize and calm the viewer. (soft music) - [Voiceover] It was one of those days where it's a minute away from snowing and there's this electricity in the air.
- Let's go to the other end of the spectrum. If composers want to convey tension or unrest, they might use heavy repetition, sharp, uneven articulation, dissonant harmonies, and non-chord tones. In another scene excerpt from American Beauty, which is also comprised of very slow, long shots, we get a totally different feeling, again spearheaded by the music. Pay attention to how the music controls the rhythm of the scene, how it makes you feel, and how the inserted moments of total silence work with this technique as well.
(upbeat ukulele music) (suspenseful music) Music also defines and shapes style. There are countless styles of film, and each one comes with its own varieties of music. If you listen to these few examples, even with your eyes closed, you'd likely know that this is a Western, (whistling Western music) this is an adventure film, (suspenseful music) and you might even know that this is a quirky indie film.
(elevator music) These styles have been created and molded over a century, and while they are certainly variations within each style, it's important to be aware that the general musical characteristics of each style's instrumentation, melodies, rhythm, pacing, and so on, are set in the audience's minds. Therefore, when filmmakers choose to deviate from a particular style, it can work, but they just need to be aware of how that deviation may set with viewers who already have certain expectations.
Speaking of expectations, music can also function in an ironic sense. Sometimes directors choose to use music that doesn't at all match the action on screen. For example, when lighthearted music accompanies something brutal. And so sometimes this can help underscore the horror of what's going on as we see here in a battle scene in the movie Gladiator. (soft music) And in Schindler's List, a Yiddish children's choir sings as the ghetto is being liquidated.
(choral music) Music functioning in this way is a relatively common device in cinema, and it functions in different ways. Sometimes for dramatic irony, sometimes satire. But almost always does it bring the viewer out of the experience just enough to be able to observe or comment on the action at hand, which is a powerful tool for sure.
I want to expand on this a bit. When the tone of music is different from the content of the scene, it can function as a tremendous narrative device. In the film There Will Be Blood, the main character gives a speech to the people of the town where he's going to be drilling oil. I'm just showing you a excerpt here but notice that it's a matter-of-fact, borderline inspirational, children-are-the-future type of rhetoric. But listen to the music that goes with it. - [Man] Men to bring their families, as well. Of course, it makes for an ever so much more rewarding life for them.
Family means children and children means education, so, wherever we set up camp. - It's not too inspirational, is it? The music exudes dread. So while the deliver of the scene suggests optimism, the music foreshadows disaster, which is of course what happens. In the next movie, we'll expand on this and talk about music as a unifier.


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The Real Life Guys ist ein deutscher YouTube-Kanal über Do-it-yourself-Projekte, der 2016 von den Zwillingen Johannes Mickenbecker (* 27. Juni 1997) und Philipp Mickenbecker (* 27. Juni 1997; † 9. Juni 2021) aus Bickenbach, Hessen gegründet wurde.

Leben
Johannes, Philipp und Elisabeth (Elli) Mickenbecker wuchsen in einem christlich geprägten Umfeld auf, in dem sie während ihrer Grundschulzeit von ihren Eltern im Heimunterricht erzogen wurden. Nur einen Teil ihrer Jugend besuchten sie öffentliche Schulen (Freie Christliche Schule Heidelberg und Schulzentrum Marienhöhe in Darmstadt), um 2016 das Abitur ablegen zu können. Die Zwillinge begannen dann ein Studium der Umweltingenieurwissenschaften, das sie nach kurzer Zeit abbrachen.

Am 10. März 2018 verunglückte Elli Mickenbecker, die an fast allen Aktivitäten der Zwillinge teilgenommen hatte, kurz vor ihrem 19. Geburtstag bei einem Flugzeugabsturz tödlich. Zwischen März und Mai 2018 legte der von den Brüdern betriebene YouTube-Kanal eine Pause ein, auf dem in diesen Monaten nur ein Song ihrer Schwester veröffentlicht wurde.

Am 2. Oktober 2020 gab Philipp Mickenbecker in einem Video auf dem gemeinsamen YouTube-Kanal bekannt, dass bei ihm zum wiederholten Mal ein Rezidiv seiner 2013 erst-diagnostizierten Krebserkrankung (Lymphdrüsenkrebs) aufgetreten sei. Nachdem Ende 2020 noch eine Fernsehdokumentation für die ARD über Philipp Mickenbecker gedreht worden war, starb er am 9. Juni 2021 im Klinikum Darmstadt an den Folgen der Krebserkrankung. Am 17. Juni 2021 wurde er neben seiner Schwester in Bickenbach beigesetzt.

Anfang Juli 2021 wurde bekannt, dass Johannes Mickenbecker – wenige Wochen nach dem Tod seines Bruders Philipp – seine Freundin Lara auf Schloss Alsbach geheiratet hat. Sie hatten sich am Heiligabend 2020 verlobt.

Ziel und Inhalt ihrer YouTube-Aktivitäten
Ziel der Brüder bis zu Philipps Tod war es, mit den Videos junge Menschen dazu zu bewegen, in die freie Natur zu gehen und Abenteuer zu erleben. Sie versuchten damit anzuregen, dass junge Leute ihre Zeit nicht nur vor dem Computer und dem Handy verbringen. Zu diesem Zweck luden sie in unregelmäßigen Abständen Videos zu verschiedenen Projekten hoch, um damit jungen Menschen Motivation und Ideen für eigene Projekte zu geben. Das Wissen für die Umsetzung haben sie sich größtenteils selbst angeeignet und anhand von YouTube-Tutorials gelernt.

Seit 2020 wurde von den Brüdern Mickenbecker zusätzlich der YouTube-Kanal "Life Lion" betrieben, wo unter anderem Philipp mehrmals über seinen Gesundheitszustand berichtete und Gedanken zu seinem christlichen Glauben teilte. Einige der Inhalte dieses Kanals entstanden unter Mitwirkung des Schauspielers Samuel Koch.

Projekte
Die Brüder führten viele Projekte durch, die auch mediale Aufmerksamkeit auf sich zogen. Unterstützt wurden sie dabei von zahlreichen Freunden. Als Ausgangspunkt vieler ihrer Projekte dienen Badewannen.

Badewannen-U-Boot
Im Jahr 2017 bauten sie aus Badewannen ein bemanntes U-Boot, mit dem sie zahlreiche Tauchgänge unternahmen. Zur Sicherheit wurden sie von Tauchern begleitet, die im Notfall hätten eingreifen können. Im U-Boot befand sich eine Pressluftflasche. Bemannt wurden Tauchtiefen bis zu 10 Metern erreicht. Bei einem anschließenden unbemannten Versuch zur maximalen Tauchtiefe implodierte das U-Boot in einer Tiefe von 12 Metern. Rückblickend bezeichneten sie dieses Projekt als ihr gefährlichstes bis zu dieser Zeit.

Fliegende Badewanne
Im Jahr 2018 bauten die Brüder eine Badewanne zu einer bemannten Drohne um und führten damit einen der ersten bemannten Drohnenflüge in Deutschland durch. Unter anderem flogen sie mit ihr zum Bäcker. Die Drohne wurde in Rücksprache mit dem Luftfahrtbundesamt auf eine maximale Flughöhe von 30 Metern ausgelegt, so dass keine Genehmigungen nötig waren. Während des Baus arbeiteten sie auch mit dem Drohnenhersteller exabotix zusammen. Die Drohne hat eine Flugdauer von ca. 6 Minuten. Sie verfügt über sechs 20-PS-Motoren, die jeweils einen Rotor antreiben, wodurch ein Gesamtgewicht von 140 kg befördert werden kann.

Baumhaus
Mitte 2018 begannen die Brüder mit dem Bau ihres "Traum-Baumhauses". Es befindet sich in Seeheim-Jugenheim in circa 7 m Höhe und besitzt drei Stockwerke.

Godzilla
Im Jahr 2019 bauten sie anlässlich des Films Godzilla II: King of the Monsters eine 20 Meter lange und sieben Meter hohe Nachbildung von Godzilla. Sie wurde anschließend für einen Streich des Comedy-Trios ApeCrime verwendet. Dazu wurde die Godzillanachbildung im Erlensee bei Bickenbach versenkt, um durch Taucher im richtigen Moment zum Auftauchen gebracht zu werden. Dabei kam es zu Problemen, weswegen die Nachbildung sank und anschließend durch einen Hubschrauber gehoben werden musste. Unterstützt wurde das Projekt unter anderem von Warner Bros. Entertainment.

U-Boot 2.0
Nachdem sie eine Million Abonnenten erreicht hatten, begannen sie, ein zweites U-Boot aus einem Gastank zu bauen.

Achterbahn
2019 bauten sie in einem geschlossenen Hornbach-Baumarkt innerhalb von 48 Stunden eine Achterbahn nur mit Mitteln aus dem Baumarkt. Die einzige Vorgabe durch Hornbach war Sicherheit als höchste Priorität. Die Achterbahn hatte zum Schluss eine Höhe von über 10 Metern, verfügte über einen Looping und endete in einem Pool. Als Wagen kam eine umgebaute Badewanne zum Einsatz.

Community-Event
Im Juli 2021 – wenige Wochen nach Philipps Tod – wurde ein "Community-Event" am Wechselsee in Biebesheim am Rhein veranstaltet. Ziel war es, ein riesiges Floß aus Badewannen und Türen zu bauen. Das Projekt wurde auch von der örtlichen Polizei und Feuerwehr unterstützt, mehr als 300 Menschen nahmen daran teil. Ursprünglich war es geplant, das Projekt an der NATO-Rampe in Gernsheim am Rhein zu veranstalten, aus Sicherheitsbedenken entschied man sich dann aber, es an einen anderen Ort zu verlegen.

Ende Juli 2021 wurde gemeinsam mit der Organisation Samaritan's Purse ein weiteres Community-Event veranstaltet. Diesmal halfen sie bei den Aufräumarbeiten nach dem Hochwasser im Katastrophengebiet Bad Neuenahr-Ahrweiler in Rheinland-Pfalz. Der Hilfseinsatz dauerte drei Tage, mehrere hundert Menschen nahmen daran teil.

Sonstiges
Im Januar 2020 waren The Real Life Guys Teil der SWR-Reportage Rollende Raritäten: Schräge Fahrzeuge Marke Eigenbau. Hier wurden sie beim Umbau eines Kindertretautos zu einem GoKart mit Elektroantrieb sowie bei der ersten Testfahrt begleitet.

2020 und 2021 waren die Brüder Mickenbecker in Talkshows zu Gast (NDR Talk Show, Nachtcafe, Stern TV). In der Sendung Deep und deutlich. Eine NDR Talk Show machte Philipp Mickenbecker im Oktober 2020 erstmals seine Krebserkrankung öffentlich.

In den deutschen Tageszeitungen Bild und Der Spiegel erschienen Artikel über Philipp Mickenbecker.

Literatur

Einzelnachweise

YouTube-Kanal
Gegründet 2016
Kultur (Landkreis Darmstadt-Dieburg)
Zwillinge


I've kept diaries and pen-and-paper to-do lists for the better part of the past two decades, so it makes sense that I'd fall in love with bullet journaling. It's both a creative hobby and a practical one, and it's very self-guided; you can spend as much or as little time on it as you want.
As I said, I'm not particularly artistic, so I didn't go into this hobby expecting any amazing results; I mostly just wanted to learn some watercolor basics, make things that my mom and friends would tell me looked good, and entertain myself. Mission accomplished!
Good for: People who took one art class in high school and weren't completely terrible at it, people who look at cute illustrations on Instagram and wonder if they could do that.
Where to start: There are loads of free tutorials on YouTube, but I'd suggest Yao Cheng's Beginning Watercolors classes on Creativebug. (Though I actually started with her intermediate class because I'd already done a watercolor class on Brit.co.) Creativebug offers a free trial and then it's $4.95/month after that, which gets you a credit for one new class each month + ongoing access to all your old classes. (You're able to keep the class from your free trial forever, though.) The class I did was really well shot, and I love how organized everything is on the back end; the video is divided into chapters and you can easily access high-res versions of the paintings Cheng does in the video if you want to take a closer look.
Note: With this and all online video classes, give yourself permission to pause them so you can practice whenever you need to, and don't feel weird about needing to rewind and rewatch things until you understand them. Depending on what kind of learner you are, you may even want to watch an entire online class all the way through before you start trying the tutorials.
Embroidery looks impressive, but making cool things is surprisingly easy. As my friend Alanna says, if you can doodle, you can probably embroider. Sure, there are true artists who take it to a whooooole other level (like this one and this one), but I've been surprised by how much I've been able to do.
Where to start: This Creativebug class is good for learning a bunch of basic stitches. You may also be able to get everything you need from YouTube, and I was able to learn other techniques (like making roses) from photo tutorials on Pinterest. The "Females Are Strong As Hell" project is based on a design created by the extremely talented Jen Riggs (check out her Etsy shop here), and was really quite easy once I'd learned to do a simple back-stitch.
I grew up watching my grandma cross-stitch and it always seemed so complicated. When I finally asked her to teach me to do it this past spring, I learned that while it can be complicated, it also can be SO EASY AND OH MY GOD WHY DID I WAIT SO LONG TO TRY IT? That "100" is the first thing I ever did and it wasn't hard at all.
Good for: The same people who might like embroidery, but especially people in that group who are perfectionists and/or who don't think they'd be good at coming up with or sketching out their own designs. And in my experience, basic cross-stitch requires a higher level of focus and concentration than basic embroidery.


A confluence of residential, retail, hospitality, entertainment, sporting events and culture define this 75-acre neighborhood that is bordered by a major vehicular artery and commuter rail station. WDG designed the first two residential buildings on the pedestrian "main street", which created an axis connecting the park with the nearby American Airlines Center. Our role as collaborator and facilitator has, once again, helped to produce a contextually appropriate, sustainable development in a major metropolitan area.