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Henri Salvador enjoyed one of the longest careers of any French cabaret artist, debuting professionally in the mid-'30s and recording -- with satisfying results -- all the way into the new millennium. For much of his career, Salvador was known for his jazzy guitar work, his comic talents, and novelty songs, and a distinct Brazilian influence in his brand of chanson. A star in France since the end of World War II, Salvador shifted into children's music for much of the '70s before reclaiming his old audience. He reinvented himself again with 2000's Chambre Avec Vue, a gentle, nostalgic collection of love songs that, thanks in part to the Buena Vista Social Club phenomenon, was reissued in America by the legendary jazz label Blue Note. Salvador was born on July 18, 1917, in Cayenne, French Guiana. His father came from Spanish stock and his mother was descended from Caribbean natives, and both had been born on the island of Guadeloupe. The family moved to Paris when Salvador was seven, and at age 11, he discovered American jazz via Louis Armstrong and Duke Ellington. He started playing the guitar, influenced primarily by the great gypsy jazzman Django Reinhardt, and at age 16 landed his first professional gig with Paul Raiss' orchestra. In 1935, he began performing with a jazz quartet at Jimmy's Bar, a popular cabaret. In addition to establishing his talents as a comedian, this exposure led to a gig with American violinist Eddy South in 1936, and also a meeting with his idol Django Reinhardt, for whom he served as accompanist for a brief period. Salvador's promising career was interrupted by World War II; he enlisted in the military in 1937 and served for four years. He quickly found work with Bernard Hilda's Cannes-based jazz orchestra, and from there was hired by orchestra leader Ray Ventura for his comic presence. Ventura's group spent much of the remainder of World War II touring South America, particularly Brazil, with an act highlighted by Salvador's vocal impressions of Popeye. He performed his first solo shows in Brazil in 1942, and when he returned to France after the war in 1945, he decided to embark on a solo career. Salvador's time in Brazil helped him break away from his dominant Reinhardt influence, and he incorporated elements of samba into much of his subsequent work. He started his own group and, in 1947, cut his first solo sides for Polydor, including "Clopin Clopant," "Maladie de l'Amour," and "Ma Doudou." The following year, he appeared in the operetta Le Chevalier Bayard alongside Yves Montand. In 1949, Salvador was awarded the prestigious Grand Prix du Disque de l'Académie Charles Cros on the strength of two hits, "Parce Que Ça Me Donne du Courage" and "Le Portrait de Tante Caroline." He followed them with one of his all-time classics, "Le Loup, la Biche, et le Chevalier (Une Chanson Douce)." Salvador spent much of the early '50s giving live performances, both in France and abroad. He moved over to the Philips label in 1952 and issued the LP À Pleyel in 1955. Late the following year, he made his first appearance in the United States, right in the midst of the rock & roll craze. After appearing on The Ed Sullivan Show, Salvador returned to France and hooked up with songwriting partner Boris Vian. Adopting the pseudonym Henri Cording, Salvador began writing rock & roll songs with Vian and recording them with a group dubbed the Original Rock and Roll Boys. Oddly humorous novelty numbers like "Rock and Roll Mops" and "Le Blues du Dentiste" gave France some of its earliest exposure to the new music. Salvador moved to the Barclay label in 1958, but unfortunately, Vian passed away the following year, having collaborated on over 400 songs with Salvador in their brief but hugely productive partnership. Salvador soldiered on, culling another live album, Alhambra, from his performances at the titular venue in 1960. He embarked on a successful 12-week run on Italian television in 1961, which encouraged him to concentrate almost exclusively on that medium for his live performances. He and his wife Jacqueline started their own music publishing company and label, Disques Salvador, and he quickly boosted them with a hit, "Le Lion Est Mort Ce Soir," in 1962. He followed that with the Monsieur Boum Boum LP in 1963, and subsequently started a new label, Rigolo, that would be the home of a series of hit singles from 1964-1968: "Syracuse," "Zorro Est Arrivé" (an adaptation of the Coasters' "Along Came Jones"), "Le Travail C'est le Santé," "Juanita Banana," "Veunise," "Quand Faut Y Aller, Faut Y Aller," and "Mon Pote le Blues," among several others. Salvador closed out the '60s with a series of popular television specials, as well as the LP C'est Beau de Faire un Show in 1969. In 1971, Salvador morphed into a children's singer with an original song that recounted the plot of the Disney film The Aristocats. The resulting album, Henri Salvador Chante 'Les Aristochats' et le Monde Merveilleux de Disney, helped him win his second Grand Prix du Disque. Over the next five years, Salvador recorded five more children's albums that relied heavily on Disney films, in particular tackling Snow White and the Seven Dwarfs, Robin Hood, and Pinocchio; he also made recordings of LaFontaine's fables. Following his wife's death in 1976, Salvador returned to adult music, issuing two albums -- Salvador 77 and Les Canotiers -- over the next two years. In 1979, he recorded Salvador/Vian, a tribute album for Boris Vian that revisited 12 of their best-known works. Salvador en Fête followed in 1980, recapping some of his most celebrated moments on record. After this burst of activity, Salvador slowed his pace a bit, concentrating more on television performances and his return to the concert stage in 1982. The latter was documented on the double-album Live du Spectacle de la Porte de Pantin. In 1984, Salvador retired his label -- in part because his wife had always handled most of its everyday operation -- and signed with EMI/Pathé Marconi, for whom he debuted with 1985's all-new Henri. The French songwriters union SACEM honored him with its Grand Prix de l'Humour in 1987, and the following year he was knighted as a Chevalier in the Legion d'Honneur. A new album, Des Goûts et des Couleurs, appeared in 1989 and proved to be his last for EMI. Salvador kicked off the '90s by returning to his roots in jazz and blues. He appeared at the Montreux Jazz Festival in 1991, and two years later performed with the great French jazz pianist Michel Petrucciani. In 1994, he signed a new deal with Sony and traveled to New York to record the jazzy Monsieur Henri album; the live Casino de Paris followed a year later. In 1996, he received a special lifetime achievement award at the Victoires de la Musique Awards, where he performed a duet with Ray Charles. After resting comfortably on his laurels for several years, Salvador returned to recording in 2000 with Chambre Avec Vue, a high-profile comeback that featured a number of promising young songwriters, a duet with Françoise Hardy, and some of Salvador's first self-penned material in quite some time. A gentle, delicate, romantic fusion of French pop and bossa nova, Chambre Avec Vue was a huge hit with the French public; it also won him Best Male Artist and Album of the Year awards at the Victoires de la Musique. The recent success of the Buena Vista Social Club album and documentary in America had suddenly made hot commodities of older foreign musicians, and in 2002, Blue Note reissued Salvador's album under its English title, Room with a View. Meanwhile, Salvador embarked on a triumphant tour of France and, later, North America. He returned with a similar-sounding follow-up, Ma Chère et Tendre, in 2003, and three years later Révérence, an album that was recorded mostly in Brazil and included duets with Caetano Veloso and Gilberto Gil, came out. Henri Salvador died at age 90 from a ruptured aneurysm at his home in Paris on February 13, 2008.
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Q: Android Studio - WebView full size I'm trying stuff out in Android studio. It's going pretty well, but just one thing I got stuck with. How do I make the WebView activity use full screen size?
http://i.stack.imgur.com/oYJoP.png

A: requestWindowFeature(Window.FEATURE_NO_TITLE);

Before the setContentView
setContentView(R.layout.yourlayout);

in Activity oncreate method

A: Use the following code. this worked for me
<WebView
android:id="@+id/webView"
android:layout_width="match_parent"
android:layout_height="match_parent">
</WebView>

If you are using WebView in another Layout such as LinearLayout don't give padding to that layout...
<LinearLayout
    android:layout_width="match_parent"
    android:layout_height="match_parent"
    android:orientation="vertical">
    <WebView
        android:id="@+id/webView"
        android:layout_width="match_parent"
        android:layout_height="match_parent">
    </WebView>
</LinearLayout>

If you want to hide ActionBar too then goto your manifest file and in your activity tag use android:theme="@style/Theme.AppCompat.Light.NoActionBar"
<activity
   android:name=".YourActivityName"
   android:theme="@style/Theme.AppCompat.Light.NoActionBar"/>

redpajama_set_name: RedPajamaStackExchange


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JoVE Journal Biochemistry
Acetylated histone proteins can be easily expressed in Escherichia coli encoding a mutant, Nε-acetyl-lysine (AcK)-specific Methanosarcina mazi pyrrolysine tRNA-synthetase (MmAcKRS1) and its cognate tRNA (tRNAPyl) to assemble reconstituted mononucleosomes with site specific acetylated histones. MmAcKRS1 and tRNAPyl deliver AcK at an amber mutation site in the mRNA of choice during translation in Escherichia coli. This technique has been used extensively to incorporate AcK at H3 lysine sites. Pyrrolysyl-tRNA synthetase (PylRS) can also be easily evolved to incorporate other noncanonical amino acids (ncAAs) for site specific protein modification or functionalization. Here we detail a method to incorporate AcK using the MmAcKRS1 system into histone H3 and integrate acetylated H3 proteins into reconstituted mononucleosomes. Acetylated reconstituted mononucleosomes can be used in biochemical and binding assays, structure determination, and more. Obtaining modified mononucleosomes is crucial for designing experiments related to discovering new interactions and understanding epigenetics.
We have utilized PylRS and tRNAPyl to synthesize and assemble reconstituted mononucleosomes with site specific acetylated histones. PylRS has proven invaluable as a genetic code expansion tool to produce proteins with post translational modifications (PTMs) and has been genetically evolved to incorporate about 200 different ncAAs. PylRS incorporates at an amber stop codon, removing competition from other amino acids during translation. PylRS was first discovered in methanogenic archaea, and has since been utilized in chemical biology to incorporate novel reactive chemical groups into proteins1,2.
MmAcKRS1 was evolved from Methanosarcina mazei PylRS and frequently used in our laboratory for the synthesis of acetylated proteins3,4,5,6. MmAcKRS1 delivers AcK at an amber mutation site in the mRNA of choice during translation in Escherichia coli. This technique has been previously used to incorporate AcK at K4, K9, K14, K18, K23, K27, K36, K56, K64, and K79 histone H3 lysine sites to study the activity of Sirtuin 1, 2, 6, and 7 on acetylated mononucleosomes4,5,6. Here we detail this method to express acetylated histones and reconstitute acetylated nucleosomes.
Subscription Required. Please recommend JoVE to your librarian.
1. Plasmid construction
Begin by deciding which histone protein will be acetylated and at which lysine site. Mutate the site to the amber stop codon (TAG) using site directed mutagenesis.
NOTE: There are four previously designed plasmids utilized for expression of histone proteins. All four histone proteins were cloned into the pETDuet-1 vector with an N-terminal histidine tag. Histone H4 also includes a SUMO tag, the origin of replication colE1 with a copy number of approximately 40, ampicillin resistant, and a T7 promoter.
Design forward and reverse primers that contain a TAG mutation at the desired site (replacing an existing lysine codon with the amber stop codon) in one of the four histone protein plasmids.
Use whole-plasmid PCR to amplify the TAG containing plasmid. Determine an appropriate annealing temperature for the primers. In general, the melting temperature (Tm) of the primers minus 5 °C will be sufficient. If difficulty is experienced obtaining a PCR product, a temperature gradient around Tm - 5 °C can be used to optimize the annealing temperature for amplification.
NOTE: In this experiment an in-house expressed PFU polymerase was used for 30 cycles with the following conditions: 94 °C for 30 s (denaturation), annealing temperature for 30 s, and 72 °C for 6 min (or 1 min per kilobase-pairs). For more details on this type of PCR method see Liu and Naismith7.
Clean up the PCR product with a PCR clean up kit by following the manufacturer's protocol. Analyze the PCR product by agarose gel electrophoresis and subsequent ethidium bromide staining.
Ligate the plasmid by incubating with T4 ligase overnight at 16 °C.
First transform the TAG containing plasmid (AmpR) into chemically competent Escherichia coli. Pick a minimum of 4 colonies from the transformation and purify the plasmid. Make cell stocks of each with glycerol using standard methods and store at -80 °C. Send for sequencing to confirm the desired TAG mutation.
Once the sequence is confirmed, co-transform the TAG-containing plasmid (AmpR) with pEVOL-AckRS (ChlR) into chemically competent CobB (the only histone lysine deacetylase known in E. coli.) deletion BL21 cells using the heat shock method. pEVOL is a plasmid with a mid-copy-number to low-copy-number p15A origin.
NOTE: The pEVOL series of plasmids were constructed based on previous studies that showed an increased expression of the orthogonal aaRS/tRNATyr pair was effective in decreasing protein truncation and in increasing overall yields of mutant proteins8. If CobB deletion cells are not accessible, proceed with chemically competent BL21 cells. CobB is a Sirtuin-like histone lysine deacetylase and Nicotinamide can be added at a 5 mM final concentration during histone protein expression to inhibit CobB as an alternative approach9.
Make a cell stock and store at -80 °C.
2. Acetylated histone protein expression
Prepare 1 L of autoclaved 2YT media (or volume of choice) in a culture flask.
Inoculate 20 mL of 2YT media containing the appropriate antibiotics with the co-transformed cell stock that has the TAG-containing plasmid (AmpR) and pEVOL-AckRS (ChlR) and grow at 37 °C to OD = 0.6 (4-6 h).
Add the appropriate antibiotics to the 1 L of autoclaved 2YT media and inoculate with the 20 mL starter culture. Grow at 37 °C to OD = 0.6-0.8 (2-3 h).
Add inducing agents and acetyllysine (AcK) to the final concentrations of IPTG 1 mM, 0.2% arabinose, and AcK 5 mM.
Grow the culture at 37 °C for 6-8 h.
Pellet cells at 2,700 x g for 15 min.
Store the cell pellet at -80 °C overnight.
From this step onwards keep the sample on ice all times. Dissolve the cell pellet in 50 mL of histone lysis buffer per 1 L of culture.
Sonicate according to the following cycle: 1 s on, 1 s off, total on time: 3 min at 60% amplitude.
Pellet inclusion bodies at 41,600 x g for 45 min.
Discard the supernatant and resuspend inclusion bodies in 30 mL of histone lysis buffer. Pellet inclusion bodes at 41,600 x g for 30 min.
Discard the supernatant and resuspend inclusion bodies in 30 mL of pellet wash buffer. Pellet inclusion bodies at 41,600 x g for 30 min.
Discard the supernatant and dissolve inclusion bodies in 25 mL of 6 M Guanidine Hydrochloride (GuHCl) buffer. Incubate with agitation at 37 °C for 1 h.
NOTE: If needed, inclusion bodies can be incubated with agitation overnight at 4 °C.
Pellet inclusion bodes at 41,600 x g for 45 min. Incubate the supernatant with 1 mL of Ni-NTA resin equilibrated with 6 M GuHCl buffer for 2 h.
Proceed with Ni-NTA purification under denatured conditions. Wash the column with 3 column volumes of column wash buffer. Elute acetylated histone protein with 10 mL of elution buffer.
NOTE: Attempting to concentrate the protein here is an option but acetylated histones are very unpredictable and can easily precipitate. It is not recommended to concentrate past 2 mL in total volume.
Extensively dialyze the acetylated histone protein against 5% acetic acid buffer to remove salts. Dialyze for 3 h at a time at 4 °C, exchanging for fresh buffer a minimum of 6 times.
NOTE: For improved protein purity, there is an option to dialyze against pure water. However, this causes significant precipitation and reduction in yield which may be undesirable for low-yielding acetylated histone proteins.
Aliquot and lyophilize protein, and store indefinitely at -80 °C. Wild type histone proteins generally yield 10-50 mg/L whereas acetylated histone proteins yield less than 10 mg/L depending on the specific lysine site. For example, H3K79Ac averages 5 mg/L.
3. Wild-type histone protein expression
To assemble full nucleosomes, express all 4 histone proteins. When expressing and purifying wild type histones the protocol is the same as the acetylated histones except the following:
Do not perform co-transformation. Do not use pEVOL-AcKRS for wild type histone expression. Adjust antibiotics accordingly.
Do not use a CobB deletion cell line or add Nicotinamide, AcK, or arabinose during induction of cellular expression.
4. Preparation of 601 DNA
NOTE: A previously designed optimized DNA sequence to direct nucleosome positioning is assembled with histone octamers to produce mononucleosome with high efficiency. This sequence is referred to as 601 DNA or the Widom sequence. This sequence has become the standard DNA sequence for in vitro studies of nucleosomes from chromatin remodeling assays to single molecule measurements10.
To prepare significant quantities of 601 DNA for nucleosome assembly, transform pGEM-3z/601 into Top 10 cells and grow 10 mL of culture for plasmid amplification and extraction. Use a plasmid extraction kit and follow manufacturer's recommendation.
Use an in-house expressed PFU polymerase (more efficient) for this amplification protocol. Set up a PCR reaction: For 250 µL reaction, use 207.5 µL autoclaved MQ H2O, 25 µL 10x PFU Buffer, 5 µL Forward Primer: ctggagaatcccggtgccg, 5 µL Reverse Primer: acaggatgtatatatctgacacg, 5 µL dNTP mix, 2.5 µL PFU enzyme. We typically run 60 reactions at once to get enough DNA to assemble.
Use an annealing temperature of 52 °C for 30 s and an extension time of 30 s if using PFU polymerase. Otherwise follow the manufacturers recommended conditions.
Once PCR is done, use a PCR clean up kit of choice to purify the PCR product.
5. Assembly of histone octamer
Dissolve aliquots of histone protein pellets H2A, H2B, H3, and H4 so that there is a separate stock of each histone protein in GuHCl Buffer with a total volume of 100 µL.
Calculate the concentration of each histone protein by measuring the A280 absorbance.
If absorbance is greater than 1 for any protein, dilute with GuHCl buffer to get a more accurate concentration.
Combine H2A and H2B proteins in a 1:1 molar ratio and dilute to a total protein concentration of 4 µg/µL. Repeat for H3 and H4.
Dialyze sequentially at 4 °C against 2 M TE buffer overnight, 1 M TE buffer for 2 h and 0.5 M TE buffer for 5 h.
NOTE: The second and third steps of dialysis causes unstable conformations of protein to precipitate out. It is expected to see heavy precipitation at these steps.
Remove precipitates by centrifugation at 16,800 x g at 4 °C.
Again, calculate the concentration of histone dimers (H2A/H2B mixture) and tetramers (H3/H4 mixture) measuring the A280 absorbance.
Mix dimers and tetramers in a 1:1 molar ratio and adjust NaCl to 2 M by the addition of solid NaCl.
Histone octamer can be stored at 4 °C and is more stable than dimers and tetramers. Never freeze histone octamer as this can cause disassembly.
If assembling with acetylated histones, simply replace the wild type protein with the acetylated protein in the procedure.
6. Nucleosome assembly
Use 100x TE buffer and solid NaCl to adjust 601 DNA to 2M TE buffer. Add the 601 DNA in 2M TE buffer to histone octamer in a molar ratio of 0.85:1 to 0.90:1. A lower ratio of DNA may be added if free DNA is present in the gel shift analysis.
Transfer the DNA-histone mixture a dialysis bag and place in about 200 mL of 2M TE buffer (or more if assembling multiple samples) and very gently stir at 4 °C.
Set a peristaltic pump to purge to slowly drip in no salt TE buffer. When the volume has roughly doubled, pour out half the volume. Do this at least 4 times in total. With a pump this can take 4-8 h depending on the starting volume. Nucleosomes form when the salt concentration is reduced to 150 mM (measured by salinity meter).
After the salt concentration is reduced to 150 mM, dialyze against a 20 mM TE buffer overnight.
Remove precipitates by centrifugation and measure the concentration of the nucleosomes by A260 reading using a plate reader.
Add His-TEV protease (TEV: nucleosome 1:30, w:w ) and incubate overnight at 4 °C to remove all histidine tags. Remove the histidine tag impurities from the nucleosome solution by Ni-NTA resin pull down.
To position the nucleosome and homogenize the sample, incubate at 60 °C for 1 h.
NOTE: For acetylated sites that are particularly unstable, this step may not be advisable.
Store nucleosomes for short term (a few weeks) at 4 °C.
For long-term storage, dialyze nucleosomes against storage buffer and store at -80 °C.
Representative Results
Dimers, tetramers, and octamers can be assessed by running a 12% SDS PAGE gel (Figure 1 and Figure 2). Here you can see that some of the acetylated tetramers have a lower yield than others (Figure 1). In fact, the closer to the core region, the lower the yield observed. This is most likely due to the acetylation interfering with the assembling of the tetramer the closer you get to the core regions. A similar affect is observed after assembling octamers (Figure 2). After assembling octamers with 601 DNA sequence, the nucleosome can be assessed using a 5% 1x TBE Native PAGE gel followed with staining with ethidium bromide (Figure 3). Before TEV digestion the nucleosome band is observed near the 10k base pair mark. After TEV digestions the nucleosome band shifts down relative to the ladder. Before thermal positioning, the observed nucleosome bands will be very broad, and possibly streak (Figure 4).
Figure 1: Histone dimers and tetramers. Representative 12% SDS PAGE gel of wild type histone dimer (lane 2) and H3 acetylated tetramers (lanes 3-9). Please click here to view a larger version of this figure.
Figure 2: Histone octamers. Representative 12% SDS PAGE gel of H3 acetylated histone octamers (lanes 2-11). Please click here to view a larger version of this figure.
Figure 3: Assembled his tagged H3K64Ac nucleosome complex. Representative 1x TBE Native PAGE gel of H3K64Ac nucleosome complex (lane 2) compared to free 601 DNA (Lane 4) visualized by ethidium bromide. Lane 3 shows H3K64Ac nucleosome after incubation at 60 ˚C for one hour. It is observed in lane 3 that all nucleosome has disassembled. Please click here to view a larger version of this figure.
Figure 4: Assembled TEV digested nucleosome complex. Representative 1x TBE Native PAGE gel of wild type TEV digested nucleosome complex, and free 601 DNA. A comparison of before and after nucleosome thermal positioning. Please click here to view a larger version of this figure.
It is essential to follow this protocol in every detail during an experiment. Nucleosomes are not very stable and much trial and error has gone into determining this protocol. It is key to remove precipitates at every step (or whenever observed) because particulates can easily interfere with the assembling processes. Always keep histone samples on ice. If nucleosomes are stored at 4 °C for too long, they can spontaneously disassemble. Be sure to check any samples by Native PAGE if stored at 4 °C for more than 2 weeks before use in any experiment. Avoid vortexing octamers and nucleosomes as this can also cause them to disassemble. If issues are encountered assembling dimers, tetramers, or octamers this is usually indicative of poor protein quality. Check the protein quality by 15% SDS-PAGE. One option if protein purity is low is to dialyze the protein against pure water. This will cause heavy precipitation and greatly reduce protein yield, but will produce a purer sample. The incorporation and nucleosomal stability of each histone acetylation site varies widely. In general, the closer the site is to the N-terminal domain, the lower the yield during histone protein expression. For nucleosome stability, the closer the acetylation site is to the nucleosome core or DNA binding sites, the less stable the assembled nucleosome. If stability issues are encountered, it is crucial to always keep the nucleosome sample on ice. It may be necessary to omit the nucleosome positioning step at 60 °C as this may cause disassembly or precipitation. A major limitation of acetylated nucleosomes is their instability. It may be impossible to preform assays at higher temperatures such as 37 °C without causing nucleosomal precipitation. If experimental procedures allow, perform them at 4 °C to prevent precipitation of the acetylated nucleosome.
This method of producing nucleosomes is particularly useful for producing modified nucleosomes for binding experiments and structure determinations. This method produces nucleosomes with higher purity and a known nucleosomal DNA sequence, which eliminates confounding variables resulting in better controlled experiments and higher resolution images. It is crucial for structure determinations that the nucleosome sample is homogenous with a known DNA sequence, otherwise it will be all but impossible to obtain high resolution images.
There is an abundance of potential applications of this method. Specifically, in the field of epigenetics. It can be incredibly difficult to obtain modified proteins. Obtaining modified proteins is crucial for probing the structure and function of the writers, readers, and erasers in the many epigenetic pathways that are poorly understood. We have previously utilized this technique to probe the accessibility of nucleosomal DNA to Pst1 digestion at the H3 acetylation sites K18, K36, and K56 by assembling each mutant octamer with a modified 601 DNA sequence that contains a Pst1 digestion site. This technique can be further modified to incorporate NCAAs other than AcK. PylRS can be engineered to incorporate a host of other NCAAs. We have also incorporated Nε-(7-azidoheptanoyl)-L-lysine (AzHeK) by amber codon in Escherichia coli for recombinant expression of several AzHeK-containing histone H3 proteins to investigate the histone deacylation activity of SIRT7 at several acetylation sites. This approach revealed that SIRT7 has high activity towards deacylation of H3K36 and is also catalytically active to deacylate H3K37. H3K36 deacylation was further showed to be nucleosome dependent and can be significantly enhanced by adding a short double-stranded DNA to the acyl-nucleosome substrate that mimics the bridging DNA between nucleosomes in native chromatin6.
This method can also be modified to produce wild type nucleosomes with no modifications which can be useful in a variety of scenarios. Our group recently published a structure in collaboration with Dr. Pingwei Li's group showing the tight binding of cyclic GMP-AMP synthase (cGAS) to a negatively charged acidic patch formed by histone H2A and H2B via its second DNA binding site11. cGAS is a dsDNA sensor that catalyzes the synthesis of a cyclic dinucleotide cGAMP, which mediates the induction of type I interferons through the STING-TBK1-IRF3 signaling axis12,13,14,15,16.
The benefit of using fully assembled nucleosomes is it more closely resembles the native state of nucleosomes serving as a better model to probe the activities or binding abilities of any number of epigenetic-related proteins. There are numerous examples of limited or entirely absent enzyme activity towards naked DNA or histone peptide substrates that when probed with nucleosome substrates drastically changes results. As with the example mentioned above, a novel deacylation site was discovered for SIRT7 by using a nucleosome substrate that would have otherwise been unknown. It is crucial to use native substrates when probing these types of systems. This technique can be used to probe the reading, writing, and erasing activity or binding capability of any modification that can be incorporated by the pyrrolysine genetic code expansion technique. The pyrrolysyl-tRNA synthetase is already natively promiscuous and has been engineered for a host of other substrates already opening the door for using this technique in any number of systems making the primary limiting factor the creativity of the researcher.
There are no competing financial interests to declare by any of the authors.
We would like to thank Dr. Wesley Wang for laying the groundwork for this protocol and his valuable mentorship. This work was partially supported by National Institutes of Health (Grants R01GM127575 and R01GM121584) and Welch Foundation (Grant A-1715).
Name Company Catalog Number Comments
0.5 M TE Buffer NA NA 0.5 M NaCl, 20 mM Tris, 1 mM EDTA, pH 7.8
1 M TE Buffer NA NA 1 M NaCl, 20 mM Tris, 1 mM EDTA, pH 7.8
100x TE Buffer NA NA
20 mM TE Buffer NA NA 20 mM NaCl, 20 mM Tris, 1 mM EDTA, pH 7.8
6 M GuHCl 6M guanidinium chloride, 20 mM Tris, 500 mM NaCl, pH 8.0
Acetyllysine
Column Wash Buffer NA NA 6 M urea, 500 mM NaCl, 20 mM Tris, 20 mM imidazole pH 7.8
Elution Buffer NA NA
Fisherbrand Variable-Flow Chemical Transfer Pump Fischer Scientific 15-077-67
His-TEV protease
Histone Lysis Buffer NA NA 60 mM Tris, 100 mM NaCl, 0.5% Triton-X100 (v/v), 1 mM PMSF pH 8.0
Ni-NTA Resin 6 M urea, 500 mM NaCl, 20 mM Tris, 250 mM imidazole, pH 7.8
PCR Clean-Up Kit Epoch Life Sicences 2360050
Pellet Wash Buffer NA NA 60 mM Tris, 100 mM NaCl, pH 8.0
petDUET-His-SUMO-TEV-H4
petDUET-His-TEV-H2A
petDUET-His-TEV-H2B
petDUET-His-TEV-H3
pEVOL-AckRS Addgene 137976
pGEM-3z/601 Addgene 26656
Storage Buffer NA NA 20 mM NaCl, 20 mM Tris, 20 mM NaCl, 1 mM EDTA, 20% glycerol, pH 7.8
DOWNLOAD MATERIALS LIST
Srinivasan, G., James, C. M., Krzycki, J. A. Pyrrolysine encoded by UAG in Archaea: Charging of a UAG-Decoding specialized tRNA. Science. 296, (5572), 1459-1462 (2002).
Wan, W., Tharp, J. M., Liu, W. R. Pyrrolysyl-tRNA synthetase: an ordinary enzyme but an outstanding genetic code expansion tool. Biochimica Et Biophysica Acta. 1844, (6), 1059-1070 (2014).
Umehara, T., et al. N-Acetyl lysyl-tRNA synthetases evolved by a CcdB-based selection possess N-acetyl lysine specificity in vitro and in vivo. FEBS Letters. 586, (6), 729-733 (2012).
Hsu, W. W., Wu, B., Liu, W. R. Sirtuins 1 and 2 are universal histone deacetylases. ACS Chemical Biology. 11, (3), 792-799 (2016).
Wang, W. W., Zeng, Y., Wu, B., Deiters, A., Liu, W. R. A chemical biology approach to reveal Sirt6-targeted histone H3 sites in nucleosomes. ACS Chemical Biology. 11, (7), 1973-1981 (2016).
Wang, W. W., et al. A click chemistry approach reveals the chromatin-dependent histone H3K36 deacylase nature of SIRT7. Journal of the American Chemical Society. 141, (6), 2462-2473 (2019).
Liu, H., Naismith, J. H. An efficient one-step site-directed deletion, insertion, single and multiple-site plasmid mutagenesis protocol. BMC Biotechnology. 8, (1), 91 (2008).
Young, T. S., Ahmad, I., Yin, J. A., Schultz, P. G. An enhanced system for unnatural amino acid mutagenesis in E. coli. Journal of Molecular Biology. 395, (2), 361-374 (2010).
Gallego-Jara, J., Écija Conesa, A., de Diego Puente, T., Lozano Terol, G., Cánovas Díaz, M. Characterization of CobB kinetics and inhibition by nicotinamide. PLoS One. 12, (12), 0189689 (2017).
Lowary, P. T., Widom, J. New DNA sequence rules for high affinity binding to histone octamer and sequence-directed nucleosome positioning. Journal of Molecular Biology. 276, (1), 19-42 (1998).
Zhao, B., et al. The molecular basis of tight nuclear tethering and inactivation of cGAS. Nature. 587, (7835), 673-677 (2020).
Burdette, D. L., Vance, R. E. STING and the innate immune response to nucleic acids in the cytosol. Nature Immunology. 14, (1), 19-26 (2013).
Barber, G. N. Innate immune DNA sensing pathways: STING, AIMII and the regulation of interferon production and inflammatory responses. Current Opinion in Immunology. 23, (1), 10-20 (2011).
Ablasser, A., et al. cGAS produces a 2'-5'-linked cyclic dinucleotide second messenger that activates STING. Nature. 498, (7454), 380-384 (2013).
Gao, P., et al. Cyclic [G(2',5')pA(3',5')p] is the metazoan second messenger produced by DNA-activated cyclic GMP-AMP synthase. Cell. 153, (5), 1094-1107 (2013).
Zhao, B., et al. A conserved PLPLRT/SD motif of STING mediates the recruitment and activation of TBK1. Nature. 569, (7758), 718-722 (2019).
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Rowlett, C. M., Liu, W. R. Site Specific Lysine Acetylation of Histones for Nucleosome Reconstitution using Genetic Code Expansion in Escherichia coli. J. Vis. Exp. (166), e62113, doi:10.3791/62113 (2020).… More
Rowlett, C. M., Liu, W. R. Site Specific Lysine Acetylation of Histones for Nucleosome Reconstitution using Genetic Code Expansion in Escherichia coli. J. Vis. Exp. (166), e62113, doi:10.3791/62113 (2020).
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redpajama_set_name: RedPajamaCommonCrawl


This exhibition explores both the history of money in biblical times, and shows how the Bible influenced the development of Christian images and symbols, particularly on coins.
Both the Old and New Testaments mention various forms of money used in everyday life. From the weighing of bullion as shekels and talents in the Old Testament to the 30 pieces of silver given to Judas as payment for betraying Jesus, the language of money is found throughout the biblical narrative. Objects featured in the exhibition include early coins from the Persian Empire and ancient Greece, coins from the Roman Empire and the Jewish kingdoms, an early ivory panel depicting the Crucifixion, and papyrus fragments and vellum featuring Bible texts.
The Bible narrative and the way the words in the Bible were first written down shaped early Christian imagery, as seen on coins from the Christian Roman Empire. Symbols known as the Christogram and staurogram were based on Greek letters (chi-rho and tau-rho). The staurogram was originally used in Biblical texts as part of abbreviated words, a convention of early Christian writings employed to add an element of mystical numerological wordplay. Both the depiction of Jesus himself and the now-familiar cross were not in common use until much later.
The original writers of the Bible, as well as subsequent translators, needed to refer to coin denominations in terms that would be familiar to readers and in sums that made sense to the stories. An early edition of the King James Version of the Bible is included in the display to illustrate the translation of coin types alongside corresponding 16th–17th-century coins.
From tax to the Temple, from shekels to silver, this exhibition reveals the fascinating yet often hidden interaction between money and the Bible.


text:
Williamsport ist die Bezeichnung mehrerer

Orte in den Vereinigten Staaten:
 Williamsport (Indiana)
 Williamsport (Kentucky)
 Williamsport (Louisiana)
 Williamsport (Maryland)
 Williamsport (Ohio)
 Williamsport (Pennsylvania)
 Williamsport (Tennessee)
 Williamsport (West Virginia)

redpajama_set_name: RedPajamaWikipedia


text:
Service, Contact and Arrival
We offer you the all-around carefree package. Trust our many years of experience as hosts. Leave the hustle and bustle behind and enjoy pure nature high above the city. Every 30 minutes, the Uetliberg train brings you to the top of the most beautiful local mountain in Switzerland. From the final station, Uetliberg, you'll arrive at the hotel after a pleasant seven-minute walk through the woods
You can travel by car to the Feldermoos parking in Uitikon-Waldegg, which is located 200 meters before the petrol station at 12 Birmensdorferstrasse.
A4/A3 highway from the north / west or Basel, Bern, St. Gallen and coming from Zurich-Kloten airport: take exit 28 Oberurdorf-Süd, then drive approximately 4.9 km towards Zurich to the Feldermoos parking lot at 47 Birmensdorferstrasse, 8142 Uitikon-Waldegg, and from there take the SZU to the Uetliberg.
A4/A3 motorway from the south / east or Zug, Lucerne and Chur: take exit 29 Uitikon and then go about 3.8 km towards Zurich to the Feldermoos parking lot.
Arrival from Feldermoos parking lot:
1. 7 minute walk by foot via the natural road: cross the footbridge, follow the steps until you reach the railway track, then go right about 300 meters to the Uitikon-Waldegg train station. The SZU (S10) train leaves from there to the Uetliberg every 30 minutes on weekdays and every 20 minutes on weekends.
2. Walk ten minutes via the paved road: From the parking lot, bear right along the road to the second gas station. Use the pedestrian underpass and go to the other side of the road. Finally, a paved road leads you through the forest to the Uitikon-Waldegg train station.
Landing in Zurich Kloten, the train is your fastest and easiest choice. Every hour, direct trains (up to 22 ) go to the Zurich main station. At the main station, change to the underground SZU (S10) on 21/22. The SZU runs weekdays every half-hour and takes just 20 to get to the Uetliberg. On weekends it runs every 20 minutes. Time schedules from the airport to the Uetliberg are available on the Internet at fahrplan.zvv.ch.
The UTO KULM hotel is located in a car-free recreation area – the Uetliberg . There are several parking facilities in and around Zurich, but fortunately you can only get to the hotel via the Sihl Valley-Zurich-Train line (SZU) or by foot. After the final station at Uetliberg, there is a 7 minute walk to the hotel. On request, handicapped guests as well as the luggage of hotel and conference guests will be collected. Please inform us one day in advance of your arrival.
When do you leave?
The train is your fastest and easiest choice. From Zurich main station, take the Sihltal Zurich-Uetliberg Bahn SZU (S10) the underground station on track 21/22. The SZU runs weekdays every half-hour and takes just 20 to get to the Uetliberg. On weekends it runs every 20 minutes. Time schedules from the airport to the Uetliberg are available on the Internet at fahrplan.zvv.ch.
Discover your spring: «Dive into the world of Valser and learn all about the natural Swiss mineral water».
Long before anyone ever put it into bottles, Valser water formed springs and lakes which people could bathe in. Water coming from a factory of nature offered an internal and external benefit. It still gushes even today from the rocks at 30 degrees. Even the Romans in ancient times appreciated the uniqueness of mineral water from the Vals. It was first called St. Peter's spring in 1622. The recent success of VALSER mineral water began in 1960 and continues to thrive.
The ticketing partner for your event
Star Ticket brings advantages for organizers and ticket buyers. Our contemporary, fully web-based ticketing connects traditional pre-sale with the latest information technology, offering services at fair and transparent prices, benefiting both sides equally.
Flexibility has two names: Print at Home® and electronic access control. Print at Home® is offered for all events and is particularly attractive to ticket buyers because any advance fees are dropped: order online, pay with e-banking and print the ticket yourself. The electronic access controls allow rapid and accurate visitor check-in, faster and safer than with visual controls.
SZU – Sihltal Zürich Uetliberg Bahn
Die SZU ist die wichtigste Verbindung zwischen Zürich und dem Uetliberg.
Rund 130 Mitarbeitende sorgen dafür, dass Sie sich tagtäglich auf die Sihltal Zürich Uetliberg Bahn SZU verlassen können und innerhalb von 20 Minuten vom Hauptbahnhof bis zur Bergstation Uetliberg (und wieder zurück) kommen
Unter dem Firmendach SZU sind die S-Bahnlinien S4 und S10 sowie die Luftseilbahn Adliswil-Felsenegg vereint.
Missing Translation
Currently there is no translation available for this webpage. You will be redirected to the german section with more information.


wife was mistress of the master.
"Where did you get that child?" I asked of the shoemaker one day.
"Whom do you mean? Bill?"
"Yes, the little fellow you call Bill."
poor house as like as not."
their pay won't keep soul and body together."
"Is he bound to you?"
"Oh yes. I never take a boy without having him bound."
"What was his mother's name?"
"I believe they called her Mrs. Miller."
"Did you ever meet with her?"
own door if she had starved to death."
"Pride? Why should she be proud?"
people, and was afraid of meeting some of them in the street."
boy. He had turned also. I saw that there were tears in his eyes.
was midwinter when I next saw him.
"Well, my little man," I said, "is any one sick?"
"Please sir, Mr. Maxwell wants you to come down and see Johnny."
"Mr. Maxwell! Do you live with Mr. Maxwell?"
saw him, and changed for the worse.
"What is the matter with Johnny?" I asked.
"I believe he's got the croup."
"Indeed! Is he very sick?"
weather as this and have on no warmer clothing than covers you now.
Come up to the stove and warm yourself--you are shivering all over.
Why did not you bring an umbrella?"
"He doesn't? But he sends you out in the rain?"
"Oh yes--always. Sometimes I am wet all day."
"Doesn't it make you sick?"
sometimes my face swells up and pains me so I can't sleep."
wear them. Thomas, one of the boys, gave me these old ones."
"Why do they hurt your feet? Are they too small?"
"No, sir, I don't think they are. But my feet are sore."
them, only they're a little snow-burnt."
can't sleep at nights sometimes for the burning and itching."
with other indications of a severe attack of chilblains.
I must put my feet in snow and let it draw the cold out."
"Yes, sir, as long as I could bear it; but it hurt dreadful bad. Mr.
Maxwell said I didn't keep them in half long enough."
get them wet so often, that they can't get well."
"What is your name?" I asked.
"Were you born in Baltimore?" I continued.
"Have you any relatives here?"
"Mr. W----?" I said, in surprise.
"Yes, sir--mother said he was my uncle."
"Is he your mother's brother?"
"Did he ever come to see your mother?"
"No, sir, he never came near us, and mother never went to see him."
home as quickly as you can, and try and keep out of the rain."
turn back and speak to his master.
shoemaker, looking up with surprise.
"He is not clothed warmly enough for such weather as this."
"You must be mistaken. He has never complained of not feeling warm."
upon and interfere with his business.
the unthinking, a warning of danger. And this I give to you now."
"Do you think I have done so in the present case?"
"I don't think any thing about it. I know you have."
"Then you think the lad warmly enough clothed?"
"If I did not think so, I would dress him more warmly."
fixed my eyes intently on the man as I spoke.
"How hot do you suppose your blood is?"
out the cold at any season."
been a boy myself, and know all about it."
"Why not in his case, doctor? I can see no reason."
his blood circulate freely enough."
boy's sake I put a curb upon my feelings.
see in most boys; and this is his misfortune, not his fault."
sister. I would then be able to act with more certainty of success.
impressive one. I will give it as briefly as possible.
evidences of attachment springing up between them.
assented with a blushing cheek.
admired the former, but loved the latter unacknowledged to herself.
count the days and hours until your answer come.
answer might have been different. Now I can only bid you forget me.
forget myself! No, I can never forget you. A week!--a week earlier?
whole heart. That will be enough. He will bid you be free."
as you use to me without dishonour."
Miller. But she laid his letters aside unanswered.
union of Anna with Miller had been consummated.
acquaintance with Anna. This he deemed imprudent, as well as wrong.
the South. She complains of your neglect, and, I think, justly.
of being happy, and that was all.
"It is high time you were married."
"Westfield, why don't you go more into company?"
"I really believe you are in love with Mrs. Miller."
object of his misplaced affection.
"Miller, a'n't you jealous of Westfield?"
"Oh yes--very jealous," he returned, in mock seriousness.
intimate with her as Westfield is with your wife."
question itself, produced an instant revulsion in Miller's feelings.
Miller's face was pale, and his lip quivered as he said this.
know it. I thought you were aware of this."
"No, I never dreamed of such a thing. Surely it cannot be true."
yield to any effort his generous feelings made to banish them.
hung up, was thrown across a chair.
contents her husband had never seen, and had never desired to see.
had noticed. They were from New Orleans.
wronged--wronged in a way for which there was no atonement.
the form of beauty and innocence it had loved so purely.
before his wife's return, he had abandoned her for ever.
meal, and enjoy a social hour afterward.
inquired if he had not been home.
more than two hours ago."
came down and told Ben to take his trunk to Gadsby's."
on terms of intimacy with me."
and to him submitted the letters he had taken from Anna's casket.
shall pay the price of my sister's dishonour. Madness!"
betrayed. But the wretch shall pay for this dearly."
preliminary explanation of the cause of the visit.
"Did you write those letters?" W----asked, with a stern aspect.
"Who dares say that I did?"
"I dare say it," returned the brother, passionately.
"It is false!" replied Westfield, emphatically.
inflammation arising from his wound.
and indignant refusal to accept the support the law had awarded her.
good many questions that I did not see proper to answer directly.
"Come round to my office and let me see them," said I.
of "uppers" in his hand--"and must be back in twenty minutes, or Mr.
and then hobbled on as fast as he could.
while, and will see that Mr. Maxwell does not give you the strap.
You must come to my office and get something done for your feet."
them with a pitiable expression on his young face.
"Let me go to the binder's first."
you return; I want to see you particularly."
was springing up in his bosom.
"And that your mother was dead?"
"I think you said that W----was your uncle?"
"Yes, sir. Mother told me that he was my uncle."
"Did your mother ever speak to you about him?"
"Then you can't tell whether he is living or not?"
"No, sir; but I suppose he is dead."
"Because I never saw him, nor heard mother speak of him."
"You are sure your name is Miller?"
"And that Mr. W----is your uncle?"
"My mother said he was."
"Did you ever see him?"
"Why don't you go, to see him, and tell him who you are?"
think of such a thing."
"And so you never went to see him?"
"What became of your mother's things after she died?"
"The woman we rented from took them all. Mother owed her, she said."
"Indeed! Where did you live?"
"Does the woman live there still?"
"Do you ever go to see her?"
"No, sir; she won't let me come into the house."
"I cannot tell. She was going to send me to the poorhouse, when Mr.
beating. I never have tried to go there since."
"What is the woman's name?"
"Her name is Mrs. Claxon."
"And she lives three or four doors from Mr. Maxwell's?"
show me the house. Your mother had some furniture in her room?"
"Yes, sir. We had a bureau, and a bedstead, and a good many things."
"Do you know what was in the bureau?"
letters in it, I think."
"Did you ever see her reading them?"
"This box Mrs. Claxon kept?"
"Yes, sir; she kept every thing."
"Why are you so desirous to have that box?"
me away from where I am now."
"You don't like to live with Mr. Maxwell, then?"
I did not question him as to the reason; that was unnecessary.
necessary to be done, or very serious consequences might follow.
from the beginning that it would not do to attempt deception.
"She died here, I believe?" I continued.
"Yes, sir; she died in my house."
"She left some property in your hands, did she not?"
_property_, why, then, she did leave property."
amount for which the things sold."
frightened. She had done no such thing, as I knew very well.
likely get yourself into trouble."
"How so, sir?" she asked, with increasing alarm.
permits no one to seize upon his effects. In the case of Mrs.
be produced. Can you produce them?"
"Y-y-yes!" the alarmed creature stammered out.
like to force it open. She handed me the box as she spoke.
the casket. "You are sure the contents have not been disturbed?"
"I trust it will be found so. I will take possession of the casket.
In a few days you will hear from me."
cause justice to be done to him.
when all accuse her as I am accused, and no hope of justice is left.
that required his attention, or felt a disinclination to leave home.
and integrity as a wife, as he would cherish the apple of his eye.
fallen from the pinnacle of happiness into the dark pit of despair!
Alas! alas! Who can tell what a day may bring forth?"
of female excellence. I was never happier than when in her company.
know the woman he has wronged."
times before, and had a slight acquaintance with him.
continued as he took the chair I handed him.
"First tell me, is the writer of its contents living?"
"No," I replied; "she has been dead over two years."
I have in my hands undoubted proofs of her innocence."
The expression of his face became agonizing.
than living with my present consciousness."
condition of the poor boy.
appearances! You have heard the melancholy history of my wife?"
whom I so deeply wronged."
"The events of the past, my dear sir," I urged, "cannot be altered.
duty, and it shall be performed as faithfully as lies in my power.
But I hope for little from that source."
mother is, that W----is his uncle."
"Are you sure of this?"
"I will go with pleasure," I returned, rising.
the child across his knees, and was beating him most cruelly.
boy away. He is bound to me by law, until he is twenty-one. Bill!
don't you dare to go."
hours for inhuman treatment of this child--of _my child_."
looked eagerly in his face.
thrilled me to the finger ends.
"Yes, William; I am your father, and I have come to take you home."
most tender, confiding, hopeful, expressive countenance.
from him and carrying of the child he had so shamefully abused.
idea of the cause which had separated his mother from his father.
ignorance of the main facts here recorded.
one fashionably-dressed lady to another.
"That pale, quiet girl, who made up dresses for the children?"
"The one I sent you."
fall; but she did not keep her engagement."
keep no more engagements of that kind."
"Not dead; but very near the end of life's weary pilgrimage."
for I want to consult your taste about a bonnet."
Mrs. Bell, "and have called to ask you to go with me."
look of unfeigned surprise in the lady's countenance.
nearly numbered. I was to see her yesterday, and found her very low.
my way to her mother's house. Will you not go with me?"
"You have never looked in the face of death," said Mrs. Lee.
There came into the face of Mrs. Ellis a sudden look of wonder.
"Are you in earnest, Mrs. Bell?"
"The mind of the girl is unbalanced."
"Don't urge me, Mrs. Bell. If I go, it will make me sad for a week.
thereto. But I do not wish to look upon death."
by the form of an angel."
any thing more terrible than death?"
for none will be experienced."
house, humble in exterior, and modestly, yet neatly attired within.
seated in the small, neat parlour.
a faint smile. "She is sinking hourly."
"But continues in the same tranquil, heavenly state?"
"Is she strong enough to see any one?" asked Mrs. Bell.
instantly played upon her lips.
the shadowy hand of the dying girl.
"I have brought Mrs. Ellis to see you. You remember Mrs. Ellis?"
you inwardly so calm, so hopeful, so confident of the morning?
real? Are you so full of heavenly tranquillity?"
they turn her thoughts too realizingly upon the "last agony?" Oh no!
waters of Jordan already touching her own descending feet.
or ask the hours to linger? Oh no!"
"Ah! but the preparation for those heavenly mansions!" said Mrs.
Ellis. "The preparation, Anna! Who may be certain of this?"
"God is love. If we would be with him, we must be like him."
"How are we to be like him, Anna?" asked Mrs. Ellis.
others. We must do the same."
garments that penetrated the inward sense of hearing?
esteems them lightly; but they are precious in the sight of God.
them is the true "Euthanasy."
just heard--we must be like God."
be like him, we must do the same."
Yes; this is the secret of an easy death, and the only true secret.
THREE SCENES IN THE LIFE OF A WORLDLING.
go. The impulse is on me. I should die of inaction here."
"And such work! Delving in and grovelling close to the very ground.
attaining wealth; and, where there is a will there is always a way."
"Contentment is better than wealth."
"No, William; it is a proverb for the wise."
gained do I purpose to return."
and fixed his eyes upon him with an earnest expression.
"Do you love wealth more than life, William?"
getting riches, then you must love money more than life."
tenderly and truly. I am going forth as well for her sake as my own.
"You will see her before you leave us?"
her this letter and this ring."
hands, gazing into each other's faces.
"Farewell, William. Think of the old homestead as still your home.
will ever be open, and its hearth-fire bright for you as of old.
he was no drone in the social hive.
morrow, he would come to bear her as a bride to his pleasant home.
glittering prize? What of the pale-faced maiden he left in tears?
Has he returned to her? Does she share now his wealth and honour?
bourne. Yet he still dwells among the living.
"Death." Over her bends a manly form. There--his face is toward you.
hope mocked him with ever-attractive and ever-fading illusions.
again he flung her passionately from him.
looked up suddenly, as if her sweet voice came floating on the air!
God! I am a man again!"
never bestow on you. Henceforth we are strangers."
the punishment he had suffered!
fruitless ambition died in his bosom.
on the darkness of his spirit: "Contentment _is_ better than wealth.
Dear brother! Dear old home! Sweet Ellen! Ah, why did I leave you?
looking into my brother's face. I feel the tight grasp of his hand.
hearth-fires bright for you as of old.' Ah! do the fires still burn?
home was a type of paradise.
"All ours. And, thank God! the little flock is yet unbroken."
recognises in the stranger his long wandering, long mourned brother.
gazing at each other, then tenderly embrace.
the familiar tones of yesterday.
"Ellen!" Here is an instant oblivion of all the intervening years.
have betrayed his deeply repented perfidy.
And here we leave them, reader. "Contentment is better than wealth."
can see as deep as most people."
is highly probable that I did see him. But what then?"


text:
From Renter to Homeowner
Charles Bergman is Director of the Esperanza Neighborhood Project for New Brunswick Tomorrow. You can learn more about him here. Stay updated on Charles' work for New Brunswick via Twitter.
In New Brunswick, the vast majority of residents rent their homes (about 80%).
Low-wage work, high rental prices, expensive housing with limited vacancy, and a deteriorating housing stock are the unfortunate reality for too many of our residents hoping to transition from renters to homeowners. To tackle these challenges, New Brunswick Tomorrow (NBT) has collaborated with partners to support first-time home buyers and develop new or rehabbed housing units.
Jose Nanez and Raudi Herrera, first-generation immigrants from Colombia and Honduras, respectively, have lived in New Brunswick for seven years. While renting apartments in converted single-family homes is common in New Brunswick, it can be complicated sharing a space with others, particularly with two young children -- as is the case with Jose and Raudi. Yet buying a home seemed an unattainable dream for them. All they've ever heard was how expensive it was, and they assumed their economic situation, working in the same factory together, wouldn't allow them to do so.
While looking for a new place to live, Jose and Raudi were introduced to Address Yourself, the affordable homeownership initiative led by NBT's partner New Jersey Community Capital. Address Yourself combines budgeting, pre-purchase, and credit counseling with access to low-cost mortgage products. Along with NBT's partnership with PRAB's Housing Coalition, these support services help aspiring New Brunswick homeowners like Jose and Raudi at every step of the process to attain their dream of homeownership.
Initially, the whole home buying process seemed very complicated to them. But, counseling in their native Spanish made the experience easier and gave them confidence that they understood the whole process.
Having lived in New Brunswick for quite a few years and now accustomed to its comfort and convenience, it was important for Jose and Raudi to put down roots here. Everything they need is nearby, including their children's schools, shops, hospitals, and even the laundromat.
As luck would have it, they were given the opportunity to purchase a home on their same block. This single-family home in the Esperanza Neighborhood of New Brunswick was renovated by the Community Asset Preservation Corporation (CAPC), a subsidiary of NJCC and another partner in NBT's Esperanza Collaborative. CAPC is a non-profit organization that stabilizes neighborhoods through the acquisition and redevelopment of vacant and abandoned properties, returning them to productive use as quality, affordable homes.
"When I saw it, I could not believe it was the same deteriorated house that my friend had pointed out to me previously," Jose said of his new home, "I feel comfortable letting my kids play all over the house because when we were renting, the apartment had spaces where it wasn't safe because there were sharp edges that stuck out. But now, this is a luxury home."
Once they found this newly renovated home, they worked closely with Integro agent Raul Cabrera and the Address Yourself team to prepare for the purchase and secure financing through their partner credit unions. Though initially the home still seemed financially unattainable, Jose and Raudi were able to utilize $10,000 in down payment assistance through Address Yourself and $50,000 in subsidy fromNew Brunswick Tomorrow with funding provided by the Neighborhood Revitalization Tax Credit (NRTC) award. This significant financial assistance made all the difference in being able to afford the home. Jose and Raudi feel relief that their combined salary can comfortably pay their mortgage without having to work overtime to pay for it.
Though they say that it is a huge responsibility, it is worth it because they now feel at ease without having to worry about the physical restrictions in an apartment or limited privacy of living with others.
"There's no words to describe what it feels like to be a homeowner. There's a pride one feels in knowing that it's yours," Jose said of living in their new home.
Since the home is newly renovated, Jose and Raudi don't have plans to upgrade the home, but they do want to personalize it. They have ideas on how they'd like to change the front façade of the home and update the patio to make it their own.
The family is overjoyed at the life they have to look forward to in their new home. To other low-moderate income and immigrant families who think homeownership may be beyond their reach, they had this to say: "Buying a home is such an important thing for a family because it's where you'll live your life. Never stop dreaming and working towards it because you can do it. Just surround yourself with the right people to help you. We are so thankful for all the help we received from Raul Cabrera, our realtor, Address Yourself, CAPC, and NBT."
Any families interested in first-home homeownership can reach out to NBT's Housing Manager, Diana Diaz, at ddiaz@nbtomorrow.org.


text:
Updated by Joanna James on Feb 11, 2020
Top 5 Cultural Experiences in Malaysia - Malaysia, Truly Cultural
Known for being a tourist hub for many things that are entertaining, Malaysia also has a few cultural sites you can explore.
Kuan Yin Temple
Asia is a place that is home to many incredible temples that span out over many years. In Malaysia you can explore a few of these shrines and temples, and of them all the Kuan Yin temple is one of the most popular temples that attracts many people from all over the world. Built by the Chinese immigrants from the ancient times, this temple was built to honour the Kuan Yin who is known to be the Goddess of Mercy.
Kampung Kling Mosque
Malaysia is also home to some beautiful mosques and of them all, the Kampung Kling is the most ancient mosque in the country and this structure was built by Indian traders who were called Klings at that time. Originally, this Mosque was built using only wood and was later converted to a brick mosque. While exploring the place, you will realise that the architecture of the establishment is a beautiful blend of Indian, Arabic, Malaysian and Chinese influences.
The Mahamariamman Temple
Yet another religious site to visit when in Malaysia is the Sri Mahamariamman Temple which is a Hindu religious site that can be found in Kuala Lumpur. This Hindu temple was designed by the architecture who belonged to the Dravidian community and the temple is truly a beautiful sight that you definitely must-see when in the City. Established in the year 1873 this is a definite must add on the list of places to visit in Kuala Lumpur if you are staying in the capital at a property the likes of Alila Bangsar Kuala Lumpur.
The Saint George Church
If you want to see an Anglican Church that belongs to the 19th Century, then you definitely must make a visit to the Saint George Church in Malaysia. Constructed by the British East India Corporation, this is one of the very few old churches in the South Asian region. This church is home to a great architectural feat as it has an English palladian which is influenced highly by neoclassical and Georgian architecture. You will also find many amazing oil paintings within the church. Sunday mornings would be a great time to see the church come completely alive while the people of the area huddle here for the mass. Do check if the church still holds mass, and if so, at what times, before you time a visit to the shrine.
Malacca Sultanate Palace
You will be able to find this palace in the Northern part of the city of Malacca. The 7 floors in this building are decorated beautifully and the legend says the pillars were made with wood only and no nails were used at all. A year later after the sultan of the time came to power, the place was struck by a bolt of lightning which destroyed the palace. Another place to visit right by the Malacca Sultanate Palace is the Malacca Cultural Museum which has displays of the various artefacts that were used during that time.
cultural experiences malaysia places to visit kuala lumpur hotel kuala lumpur luxury hotels malaysia


Brain Implant Works Better than Medication- but what are the side effects?
"Patients having deep brain stimulation (DBS) are fitted with a neurostimulator, a device similar to a heart pacemaker, which is connected to electrodes placed in certain parts of the brain.
The electrical implant is connected to a small battery under the skin in the person's chest or abdomen to generate small electrical signals to stimulate the brain.
This is controlled by a hand-held device which can be switched on and off.
When switched on, the patient benefits from the blocking of abnormal nerve signals which trigger the disabling symptoms.
The operation costs around £30,000 and is currently only considered on the NHS for patients with very advanced disease.
The latest trial using a DBS system made by US firm Medtronic is the first to look at patients with early symptoms.
After two years, those having surgery and drugs had a 26 per cent improvement in quality of life compared with drugs alone and a range of other advances."
Newer:A tiny chip could stop your heart failing...Older:Free wi-fi? Free customer data?


text:
Sitifis (), era un'antica città romana, situata nell'odierna Algeria nordorientale, dove si trova Sétif, città dell'Algeria. Venne fondata, insieme a Cuicul e Mopht, come colonia con il Colonia Augusta Nerviana Martialis Veteranorum Sitifensium fra il 96 ed il 98 durante il regno di Nerva, poi divenne capitale della Mauretania Sitifense all'epoca della Tetrarchia di Diocleziano.

Collegamenti esterni

Città romane in Algeria
Sétif


text:
Q: Line bearing for wind direction for plume of smoke QGIS I'm trying to draw a line representing direction of a plume of smoke for an emergency planning scenario in QGIS. The line needs to be 5km in length and have a bearing of 85 degrees. The closest I have found to achieve this is the CAD like drawing plug-in but this uses angles rather than compass bearings, so 180 degrees would be pointing west rather than south and vice verse.
Is there a tool to draw lines using compass angles and ideally length?
Thanks.

A: Yes, the Azimuth and Distance plug-in will do this.


text:
Dozens displaced after overnight fire at Crystal Lake apartment
Police say father threw son into Atlantic Ocean to 'teach the child how to swim'
CPD and CFD play baseball for a good cause on South Side
Police chase the checkered flag for Special Olympics of Illinois
Legendary Chicago DJ and actress LaDonna Tittle discusses role in "The Chi"
Chicagoans rally against Puerto Rico governor after lewd text messages
Jakes Takes
Backseat Rider
Conversations in Health
CW50 Chicago
FOX 32 Staff
2 pedestrians critically hurt by driver in Rogers Park
(Photo by Joshua Lott/Getty Images)
Posted May 17 2019 09:40PM CDT
CHICAGO (Sun-Times Media Wire) - A man and woman crossing the street Friday were struck and critically injured by a driver in Rogers Park on the North Side.
The driver was attempting to overtake two vehicles on the right side of the street in the 6600 block of North Ridge Boulevard when they struck the pedestrians, Chicago police said.
A 23-year-old woman suffered a broken leg, as well as injuries to her head and face, police said.
A man, 30, also had injuries to his head and face, police said. Paramedics were called at 6:06 p.m. and took them to St. Francis Hospital in Evanston in critical condition.
The driver was given two tickets for striking a pedestrian in the roadway, a one ticket for overtaking a vehicle on the right, police said.
Chicago police are investigating.
Air conditioners, condensers stolen in Bronzeville: police
Posted Jul 17 2019 12:31PM CDT
Updated Jul 17 2019 12:56PM CDT
Bronzeville residents are on alert after several air conditioner and condenser units were stolen from homes in the South Side neighborhood, according to Chicago police.
The thefts happened:
Three males are suspected in these thefts, police said, but no description was provided.
Aurora man gets 30 years for being armed habitual criminal
A man has been sentenced to 30 years in prison for having a gun while on parole in 2015 in west suburban Aurora.
Judge Katherine D. Karayannis handed down the sentence July 12 to 34-year-old Ronald Smith, according to the Kane County state's attorney's office. A jury previously convicted Smith of a felony count of being an armed habitual criminal.
"Mr. Smith received the sentence he and the community deserved," Kane County State's Attorney Joe McMahon said in a statement. "He has a horrible criminal record and is a significant public safety threat."
Naperville man charged with damaging homes, cars of ex-girlfriend's family
Posted Jul 17 2019 11:14AM CDT
Updated Jul 17 2019 11:15AM CDT
A man has been charged with vandalizing the homes of his ex-girlfriend's relatives earlier this year in the west suburbs.
Colin Herbstritt, 22, is charged with five misdemeanor counts of criminal damage to property, according to a statement from Naperville police.
Between May 22 and June 19, Herbstritt damaged vehicles and homes in Naperville and DuPage County owned by his ex-girlfriend's family, police said.
Colin Kaepernick's Nike commercial is nominated for an Emmy Award
Jobs at Fox 32


SURAT: More than 2 lakh CNG users in Surat, Ankleshwar and Bharuch will have to get their cars and autos tuned for air-fuel ratio as the quality of natural gas coming to the CNG filling stations is 'rich' in hydrocarbons. Official sources said Gujarat Gas Company Limited (GGCL) is receiving unprocessed natural of rich gas from its supplier due to a rupture in a gas pipeline in Narmada during the recent floods. It is the content of hydrocarbons that decides whether natural gas is rich or lean. If the hydrocarbon content is heavy, the gas is said to be rich and if the content is mainly methane, the gas is lean. The difference between them is because of their calorific value. Normally, CNG stations receive natural gas which is lean in nature after the heavy hydrocarbons have been removed by processing. The problem arising is that unless the CNG vehicles using this rich gas are not tuned to utilize the rich gas for firing their engines, there would be technical problems. Experts said GGCL is getting this rich gas as the pipeline carrying the gas across Narmada to a processing plant at Vadodara got ruptured. While, GGCL or its supplier is not confirming the time frame for correcting this problem, sources said that the pipeline rupture may take anywhere between six to eight months to be repaired and re-commissioned, especially if it crosses a river like Narmada. "The vehicles have to be tuned for air-fuel ratio through a regional transport office approved retrofitter. There will not be any significant impact on the engine, but the tuning is recommended if the utilization of rich gas is to continue for a longer period of time," a CNG retrofitter Rakesh Patel said.


One thing classical music organizations like Rockport Music have learned is that educating an audience — especially younger audiences — is as important as getting them to buy tickets. And audiences that already love the music are always hungry to learn more about their favorite artists and repertory.
One thing classical music organizations like Rockport Music have learned is that educating an audience — especially younger audiences — is as important as getting them to buy tickets.
With cutbacks in public school music education stretching back decades, more and more people need to learn the basics of what makes music great. And audiences that already love the music are always hungry to learn more about their favorite artists and repertory.
In conjunction with the monthlong Rockport Chamber Music Festival, Rockport Music is offering almost a dozen events that appeal to families and friends of chamber music. Many are free, and all are welcoming.
Stephanie Woolf, education coordinator for the organization, is in the thick of it, scheduling programs and outreach all year long, many in the Rockport schools, and organizing events in June that have a particular focus on the music being performed.
"We like to introduce, interact and inspire," she says. "Those are our unofficial goals. We never talk down to people, especially not children, but we are always trying to enhance the concertgoing experience.
There is a wide variety of offerings this month, some aimed at families, some specific to a given concert, and some more general presentations about music or music history. As part of the four preconcert dinner conversations, Boston Conservatory musicologist Elizabeth Seitz, a regular lecturer before Boston Symphony Orchestra audiences, will discuss that evening's program on Friday, June 5, focusing on the music of Beethoven, Brahms and little-known Russian composer Reinhold Gliere.
"The most important thing is to meet the audience halfway," Seitz says. "I'm trying to create a map of what you're going to hear and what to listen for. If people have just a few signposts of what to expect, they can have what I called a 'guided listening' experience.
The Biava Quartet performs during week three of the festival, and group members figure prominently in the extra-musical activities as well. The ensemble repeats its very popular interactive family program on Saturday, June 27. "Music in Opposites" takes the universal understanding of principles like fast/slow and loud/soft to explore the deeper concepts that underlie musical works.
And on Wednesday evening, June 24, the group's cellist, Jason Calloway, will discuss the music of Mendelssohn, particularly that composer's controversial reception in the music world. This year marks the 200th anniversary of his birth, and like many organizations, RCMF has programmed a substantial amount of his music. For Calloway, it's about time.
"It's really only in my lifetime that his music has begun getting the recognition it deserves," he says. "During his lifetime, Mendelssohn was — bar none — the most acclaimed composer in the German speaking world, if not all of Europe. It was well known that his family was Jewish — his grandfather, Moses Mendelssohn, was one of the most famous philosophers of the time — but really it wasn't that important to him or to his music.
For more details, visit www.rockportmusic.org or call 978-546-7391.


text:
Questa pagina raccoglie le informazioni riguardanti il Gallipoli Calcio nelle competizioni ufficiali della stagione 2006-2007.

Rosa

Risultati

Serie C1

Girone di andata

Girone di ritorno

Coppa Italia Serie C

Bibliografia

2006-2007


UNDER CONSTRUCTION PLEASE DO NOT USE THE FOLLOWING FORM MAILER YET!
If you are interested in looking for a propety and would like for us to find you the perfect one please fill out the following form and we will get back with you as soon as possable.
For Information Regarding Callender Realty Please Contact Jan Allen .
Callender Realty, 701 N.Center St, Bonham, TX 75418.
© 2005-07 Callender Realty & Edman Designs, All Rights Reserved.
Welcome to www.Callenderrealty.com, the most comprehensive web site available for real estate in Fannin County - Bonham, Texas area. Homes For Sale, Real Estate, Realtor, Agent, Real Estate Agent, Real Estate Broker, Relocation Home, Homes, Houses, Real Estate, Realtor, Realtors, Agent, Agents, Broker, Brokers, Relocation, Moving, Bonham, Texas, Fannin, Grayson, Lamar, Hunt, Collin, County. The brokers, agents, and staff members at Callender Realty are committed to not only providing quality service but to be trendsetters by ensuring that the public as a whole and our clients and customers know that their needs are put first in every phase of the home buying/selling process.We want everyone to see that the Texoma area of North Texas has a great deal to offer and is a wonderful place to live. We are dedicated to doing our part to make this an even more fantastic community.Callender Realty owners and agents are virtually unmatched in their experience and expertise in the real estate industry with many years of working knowledge of commercial, residential, as well as land transactions in and around the DFW Metroplex area.Our office is located in a lovingly restored 1916 Arts & Crafts home near downtown Bonham Texas. It shows our love of "home" when you walk in the door.The broker/owner of Callender Realty is Jan Allen. She and her husband, Lee moved to Bonham in 1998 and opened their mortgage quality control auditing company, Callender Mortgage Services, here at that time. This business as well as their Mortgage Brokerage business also share this office space.Lee and Jan have made Fannin County their home and are involved in many church, civic, and community organizations.Callender Realty wants to be know as a company with "small town values with big city expertise".We invite you to stop by anytime and have a cup of coffee and visit. We look forward to working with you.
Don't miss this charmer! 3bed & 2bath on oversized corner lot. Well laid-out older home, newly renovated. New carpet, paint, cabinets, etc,& ROOF! New wood floor in living room and dining area. Large producing pecan tree in the front, covered deck in the back. Back yard is fenced in two sections. Garage split between parking for 1 car and large storage,work area.
Edman Designs - Let Websites Work For You! (903)640-3411.
Graphic Creations - Contact Jeremy Jones Today!
Zion Kennels - Dog Training and Boarding- Indoor Kennels and Huge Dog Play Areas!


If you're looking for high quality and personal service, you've come to the right place. At DLB Cleaning we'll give you the attention and personal service you'll come to expect and enjoy.
We offer the best in residential cleaning & move out cleaning. Our rates are competitive, with no hidden fees. We are friendly and will be there to help you every step of the way.
DLB Cleaning has been located in Colorado Springs since 2010. Let us put our experience to work for you.


text:
Q: Firebase deploy not giviing a webhook URL I am using Dialogflow to create an action for the Google assistant. I am following this tutorial: https://medium.com/voice-tech-podcast/get-current-location-of-a-user-using-helper-intents-in-actions-on-google-19fe9a8ea99f
When I use the command firebase deploy in step 5, this is the output that I get:
=== Deploying to 'location-tracker-xxxx'...

i  deploying functions
Running command: npm --prefix "$RESOURCE_DIR" run lint

> lint
> eslint .

✔  functions: Finished running predeploy script.
i  functions: ensuring required API cloudfunctions.googleapis.com is enabled...
i  functions: ensuring required API cloudbuild.googleapis.com is enabled...
✔  functions: required API cloudbuild.googleapis.com is enabled
✔  functions: required API cloudfunctions.googleapis.com is enabled
i  functions: preparing functions directory for uploading...

✔  Deploy complete!

Project Console: https://console.firebase.google.com/project/location-tracker-xxxx/overview

I am supposed to get Webhook URL which I can use as webhook URL in Dialogflow.
This is my index.js:
const functions = require("firebase-functions");
const { dialogflow, Permission, SimpleResponse } = require("actions-on-google");

const app = dialogflow();

app.intent("Default Welcome Intent", conv => {
    conv.data.requestedPermission = "DEVICE_PRECISE_LOCATION";
    conv.ask(new SimpleResponse('Welcome to location tracker'));
    return conv.ask(
      new Permission({
        context: "to locate you",
        permissions: conv.data.requestedPermission
      })
    );
  });

I have read this Stackoverflow but it did not solve my problem because there is not shown what the solution was science the code referred to is not available anymore.
Thanks!

A: As Doug alluded to in the comments, you need to ensure that you have a function exported in your code. This can be added to the bottom:
exports.ActionsOnGoogleFulfillment = functions.https.onRequest(app);

The ActionsOnGoogleFulfillment is the name of your function. It will be deployed when you run that command.
You can view all functions in the Firebase console, and you should expect it to be in the format:
https://us-central1-**PROJECT_ID**.cloudfunctions.net/**FUNCTION_NAME**


This action, filed Jan. 31, 2017, challenges President Trump's Jan. 27, 2017 Executive Order ban on admission to the U.S. of nationals of Iraq, Iran, Libya, Somalia, Sudan, Syria and Yemen. The complaint alleges that in preventing the plaintiff, a Syrian national with a valid U.S. visa, from returning to the U.S. pursuant to the EO, the defendants violated the First Amendment Establishment Clause, Fifth Amendment due process and equal protection rights, the Immigration and Nationality Act, the Administrative Procedure Act, and international law. The case was filed in the U.S. District Court for the Northern District of Illinois, seeking declaratory and injunctive relief.
The plaintiff, represented by private counsel and Prof. Bernard Harcourt of Columbia Law School, was a physician resident in the University of Illinois Chicago/Advocate Christ Hospital. He returned to the United Arab Emirates to get married on Jan. 23, and upon returning to the U.S. to continue his residency, he was detained at Abu Dhabi International Airport on Jan 29. Despite his valid visa, he was detained pursuant to the EO. If the plaintiff could not return to the U.S., he faced the risk of being deported to Syria, where he is a citizen but has never been a resident.
That same day, the plaintiff also filed a motion for a temporary restraining order and preliminary injunction. The case was assigned to Judge Elaine Bucklo. At a status hearing on February 1, the parties told the judge they had reached a "settlement in principle." Judge Bucklo set a status hearing for February 9 at which they could update her on the progress towards settlement or otherwise.
On February 2, according to news reports, the plaintiff successfully entered the United States at Chicago O'Hare International Airport. On February 8, the plaintiff filed a motion for voluntary dismissal, with leave to reinstate on or before August 1. Judge Bucklo on February 9 granted the plaintiff's motion and terminated all pending dates and motions as moot, but noting that the plaintiff still had leave to reinstate on or before August 1, 2017.
Only the complaint is publicly available for this case.


Her Royal Highness Paduka Seri Pengiran Anak Isteri Pengiran Anak Sarah binti Pengiran Haji Salleh Ab Rahaman yesterday morning joined the Brunei Open Orienteering Competition. The competition was held at the Bumiputera Complex in Bangar Town, Temburong.
Also participating were His Royal Highness the Crown Prince's son Yang Teramat Mulia Pengiran Muda 'Abdul Muntaqim as well as members of the royal family.
Organised by C.A.R.E Association, it was participated by 40 people from 5 countries, namely Brunei Darussalam, Malaysia, Republic of Singapore, Germany and the United Kingdom.
The recreation sport required a good balance between physical and mental coordination. Participants needed to find signs from one control marker to another guided by maps and compass directions within a set time.
Her Royal Highness Paduka Seri Pengiran Anak Isteri presented prizes to the winners. Malaysia's UITM team emerged as overall champion.


text:
John Carr of England 1600's
Thomas Carr I, England and Virginia
Dabney Carr
The Carr Who Is Buried Next To Thomas Jefferson at Montecello
Joseph J. Carr
Friday, March 12, 1773 was a turning point in American history.
For the previous several years relations between the American
colonists and Great Britain had steadily deteriorated. The Stamp
Act of 1765 brought "taxation without representation," while the
Townshend Act of 1767 further burdened ostensibly free colonists
with "legislation without representation." In June 1772, an
incident in Rhode Island added fuel to the simmering cauldron. The
British schooner GASP�E was burned off Newport. In response, the
British Parliament passed an act that allowed colonists to be
shipped to England for trial. The freedoms which the colonists
cherished so dearly were in terrible jeopardy.
Sensing a severe threat to colonial liberties, several prominent
Virginians elected members of the House of Burgesses   secretly met
together in Raleigh Tavern (Williamsburg) on March 11, and proposed
formation of a network of Committees of Correspondence that would allow
the colonies to keep in touch with each other, and to monitor British
intentions. Several of the burgesses in the meeting   Thomas Jefferson,
Patrick Henry, Richard Henry Lee, Francis Lightfoot Lee, and DABNEY CARR
(possibly others also) decided to offer the idea of the Committees of
Correspondence to the assembled house. The idea seems to have been Richard
Henry Lee's, with Thomas Jefferson writing the text of the formal resolution
that would be offerred for vote. But it was 29-year old lawyer Dabney Carr
who was tasked to rise in the House of Burgesses and introduce the resolution.
The resolution was passed (although not without debate), and
Carr, along with ten others, were appointed to the colonies' first
Committee of Correspondence. By February 8, 1774 only one of the
remaining twelve colonies had not established their own Committees
of Correspondence; by September 5, 1774 the first Continental
Congress met in Philadelphia ...and as they say, "the rest is
history."
The road to the American Revolution was surveyed by Dabney Carr,
but he unfortunately did not live to trod upon it. On May 16, 1773,
only two months after delivering the speech that resulted in
formation of the Committees of Correspondence, and ultimately to the
Continental Congress and the American Revolution, the youthful Dabney
Carr died of fever in Charlottesville, VA. He is buried at Monticello
(Photo A) in the Jefferson family cemetery on the southwestern slope
of Mr. Jefferson's beautiful mountain.
When you visit Thomas Jefferson's home, Monticello, the
gravesite is behind, and down the hill from, the west garden. If
you stand on the porch of Monticello, the gravel path down the hill to
the Jefferson family gravesite is on the left. Follow the path along
the west lawn, and then down the hill to the gravesite, which is
guarded by a wrought iron fence. Follow the path around to the right,
along the edge of the site, to the gate (it's locked). Thomas Jefferson's
tombstone is right inside the gate. Immediately to the right of Jefferson's
own tombstone is that of Dabney Carr. The bronze plaque at the site reads
(in part) as follows:
This graveyard had its beginning in an agreement
between two young men, Thomas Jefferson and Dabney Carr,
who were school-mates and friends. They agreed that they
would be buried under a great oak that stood here.
Carr, who married Jefferson's sister, died in 1773. His
was the first grave on this site, which Jefferson laid
out as a family burying ground. Jefferson was buried here
According to the story, Jefferson and Carr had studied under
the "great oak" while school-mates. They both loved the location, and
pledged that whichever died first would bury the other under the tree. Only a dozen or so years later Dabney Carr would be the first buried there, as Thomas Jefferson kept his youthful promise.
Dabney Carr was born on October 26, 1743 at a thousand-acre
Louisa County, Virginia farm named Bear Castle. He was the son of
John Carr, grandson of Major Thomas Carr, and great-grandson of
"Thomas Carr, Gentlemen," who held extensive land patents in
Virginia from about 1701. Dabney attended the academy of Reverend
James Maury. Other students at the prestigious private school were
Thomas Jefferson and Matthew Maury. At the age of eighteen, Dabney
Carr enrolled in William and Mary College in Williamsburg, and
later studied to be a lawyer. Although his legal education
("reading law" under a practicing attorney) was interrupted in 1763
by militia service on the frontier with the Louisa County Volunteer
Rangers, Dabney was licensed to practice law only two years after leaving
college. In July 1765, Dabney Carr married Martha Jefferson, Thomas'
sister. The couple made their home at Spring Forest in Goochland
County, VA.
Dabney Carr was elected to the House of Burgesses in 1771 and
1772, and served on two House committees, including the influential
Committee of Privileges and Elections. He helped incorporate the Virginia
Society for the Promotion of Useful Knowledge, a group "...dedicated to
a discussion of geography, natural history, natural philosophy,
agriculture, practical mathematics, commerce, medicine and American history."
Contemporaries regarded Dabney Carr as a powerful orator who was a
serious challenge to the acknowledged master orator, Patrick
Henry. Of Carr, Patrick Henry's biographer, William Wirt, said
[Dabney Carr] "...was considered...the most formidible rival in
forensic eloquence that Mr. Henry had ever yet had to encounter."
Of Carr, Thomas Jefferson said he "...was one of the earliest and
most distinquished leaders in the opposition to British tyranny."
Although Dabney Carr is largely forgotten by the history books,
except for a little microbe ("bilious fever" the doctor called it)
he would surely have been one of the giants of the American
Revolution. As it was, Dabney Carr's contribution to the formation
of the American democracy is subtantial, even though he died young.
Notes on the Family Line of Dabney Carr
Descendents of Virgnia Carrs from the family of Dabney Carr
often become confused and erroneously append their ancestors to the
Loudoun County line (the opposite mistake is also made quite
frequently, i.e. descendents of Loudoun Carrs appending their kin
to the line of Dabney. To serve the former, I am including the
material that I have on the "downstate" Carrs related to Dabney
Carr.
These families are found largely in Albemarle County, Louisa
County, Caroline County and in the Shenandoah Valley Counties of
Virginia stretching out as far south as North Carolina. A person
with the surname Carr from these counties is not necessarily a
relative of the family described below, but their numbers are
sufficiently large to warrant looking at that connection. Keep in
mind that the lines of John Carr in Loudoun, and relatives of
Dabney Carr downstate, are not the only Carr families existing in
Virginia at the time. So please don't be too quick to append your
own line to another without documentary evidence.
Thomas Carr. (b.1655, d. >1724). Emigrated from England to
Topping Castle, Caroline County, Virginia in late 17th century.
Thomas Carr of Louisa County, VA. This Thomas Carr is styled
"Thomas Carr, gentleman" in a patent granted to him on April 25,
1701 for 546 acres of land in St. John's Parish, Pamunkey Neck,
King William County (VA) "...for the transportation of 11 persons
in the colony...." Positions held by Thomas Carr: Justice (1702),
High Sheriff (1708-09). Genealogies of Virginia Families Vol. I, p. 588ff,
reprints a letter from Col. Wilson Miles Cary of Baltimore, written
to The William and Mary College Quarterly Historical Magazine, who
asserts that Thomas Carr had two sons: Major Thomas Carr of Caroline
County (VA), born 1678, and William Carr (who is the ancestor of the
Carrs in Spotsylvania County, VA, and also of Kentucky and Missouri).
Major Thomas Carr was appointed Justice for Caroline County at
the time of its formation in 1728; he served in the Caroline court
until his death (May 29, 1737). He has also been Justice in King
William County from 1714, and was High Sheriff in 1722-23.
Major Thomas Carr was married to Mary Dabney (b. 1688, d. Sept.
7, 1748) in 1704. The children of this marraige were: Thomas (b.
1705, d. 1743 without children); John (below); Agnes (married in
1730 to Colonel John Waller of Spotsylvania); Sarah (b. Nov. 14,
1714, d. 1772), who married John Minor (1702-43), a wealthy planter of
Spotsylvania County.
The son of Major Thomas Carr who left descendents was John Carr,
Esq. of "Bear Castle," Elk Run, Louisa County, VA.; He was born on
December 26, 1706 in Caroline County, VA. John Carr, who owned extensive
estates, was a Member of the County Court (Louisa) from its formation
in 1742 until his death, and served as High Sheriff in 1753-54.
John Carr, Esq. was married twice. His first wife, Mary Garland,
died March 10, 1736. Their son was Thomas Carr (b. Nov. 24, 1735), is
recognized as the direct ancestor of the Carr line in North Garden,
Albemarle County. The second wife of John Carr was Barbara Overton
(b. April 20, 1720, d. Dec. 1794), daughter of Capt. James Overton of
Hanover. John and Barbara had eleven children, of which six survived: Dabney (discussed below); Samuel (b.1765, d.1777); Elizabeth (b.1747); Overton (b. 1752), married Ann Addison of Oxon Hill, MD.; Garland (b.1754, d.1837), of Albemarle County, married Mary Phillips (nee' Winston) in 1783; Mary (b. 1756).
William, the other son of the immigrant Thomas Carr and brother
of Major Thomas Carr, died in 1760 sometime between the making of
his will (dated Aug. 2, 1760) and its "proving" in Spotsylvania
county on August 12, 1760. His wife was named Susannah, and their
children were: Sarah Carr (married Mordecai Hord), Thomas Carr,
William Carr, Ann Carr, Elizabeth Carr, Phebe Carr, Walter Chiles
Carr, Charles Brooks Carr, Agnes Brooks Carr (married William
Ellis), Susannah Carr, Mary Carr.
Walter Chiles Carr (d. 1848) married Elizabeth Chiles. Their
children were: Susan Carr, Charles Carr, Phebe Carr, Thomas Carr,
Nancy Carr, William Chiles Carr, Virginia Carr, Dabney Carr, Walter Carr, Eliza Minor Carr, and Hulda Carr.
Charles Carr, son of Walter and Elizabeth (Chiles), married
Elizabeth Todd of Fayette County, Kentucky. Elizabeth was the
daughter of Levi Todd, and sister of Robert(?) Todd (who was the
father of Mary Todd, wife of Abraham Lincoln). Of the thirteen
children of Charles and Elizabeth, Robert Elisha Carr married Sarah Block
(who was Jewish).
Dabney Carr, grandson of Thomas Carr and son of John Carr,
married Martha Jefferson, sister of Thomas Jefferson, on July 20,
1765. Dabney Carr served in the Virginia House of Burgesses and was a
leader in pre-revolutionary Virginia. He died on May 16, 1773 of fever,
and is buried in the Jefferson family cemetery at Montecello
(Charlottesville, VA).
William Carr and Susannah Brook Carr Family Line
1. William Carr Capt. (1707-1760) & Susannah Brook (1711-)
|    1.1 Mary Carr (1730-) & Nicholas Crenshaw (1730-)
|    |    1.1.1 William Crenshaw (1755-)
|    |    1.1.2 Elizabeth Crenshaw (1757-) & Thomas Whitelow
|    |    1.1.3 John Crenshaw (1759-1818) & Mildred Thompson (-1834)
|    |    1.1.4 Ann Crenshaw
|    |    1.1.5 Susannah Crenshaw
|    |    1.1.6 David Crenshaw (1771-1831)
|    |    1.1.7 Sarah Crenshaw
|    1.2 Sarah Carr (1732-) & Mordecoi Hord
|    1.3 Susannah Brook Carr (1707-1800) & William A. Crenshaw (1725-1799)
|    |    1.3.1 Cornelius Crenshaw
|    |    1.3.2 Charles Brook Crenshaw (1749-1814) & Eunice White
|    |    1.3.3 David Crenshaw (1751-) & Jane Waddy
|    |    1.3.4 Nancy "Ann" Crenshaw (1756-1800) & Thomas Crenshaw
|    |    1.3.5 Mary Crenshaw (1762-) & John Crenshaw
|    |    1.3.6 Susannah Crenshaw (1767-) & John Burnly
|    |    1.3.7 Agnes "Aggy" Crenshaw (1769-) & Thomas Fretwell
|    |    1.3.8 Jamima Crenshaw  & William Fretwell Jr. (1736-1807)
|    |    1.3.9 Joel Crenshaw (1776-) & Jane (Jean) Swift
|    |    1.3.10 Sarah Crenshaw (1778-) & William Dickinson
|    |    1.3.11 Patsy Crenshaw (1780-) & Hardin Turner
|    1.4 Thomas Carr
|    1.5 William Carr
|    1.6 Ann Carr
|    1.7 Elizabeth Carr
|    1.8 Phebe Carr
|    1.9 Walter Carr
|    1.10 Charles Brook Carr
|    1.11 Agnes Brook Carr
Carr Mail List and Websites
CARR. A mailing list for the discussion and sharing of information regarding the Carr surname and variations (e.g., Kerr, Karr, Corr, Car, Kar) in any place and at any time. Mailing address for postings is CARR-L@rootsweb.com. To subscribe send the word "subscribe" (without the quotes) as the only text in the body of a message to CARR-L-request@rootsweb.com (mail mode) or CARR-D-request@rootsweb.com(digest mode).
Joseph Carr, 1790, PA http://www.familytreemaker.com/users/c/a/r/David-R-Carr-Illinois/index.html
Carr and King https://www.angelfire.com/de/idonotknow
Carr, Karr and Kerr Home page http://www.hctc.com/~hamkarr/index.htm
Let me add your Carr URL page here. Please add a link to my page. Thank you.
You are visitor # as of 1 June 1998.