Cloning of actin genes from a genomic library of the newt, Notophthalmus viridescens

The regenerating urodele limb is a useful model system in which to study, in vivo, the controls of cell proliferation and differentiation. Techniques are available which enable one to experimentally manipulate mitogenic influences upon the blastema, as well the morphogenesis of the regenerating 11mb. Although classical regeneration studies have generated a wealth of knowledge concerning tissue interactions, little 1s known about the process at the level of gene expression. The aim of this project was to clone potentially developmentally regulated genes from a newt genomic library for use in future studies of gene expression during limb regeneration. We decided to clone the cytoskeletal actin gene for the following reasons: 1. its expression reflects the proliferative and differentiatlve states of cells in other systems 2. the high copy number of cytoplasmic actin pseudogenes in other vertebrates and the high degree of evolutionary sequence conservation among actin genes increased the chance of cloning one of the newt cytoplasmic actin genes. 3. Preliminary experiments indicated that a newt actin could probably be identified using an available chick ~-actln gene for a molecular probe. Two independent recombinant phage clones, containing actin homologous inserts, were isolated from a newt genomic library by hybridization with the chick actin probe. Restriction mapping identified actin homologous sequences within the newt DNA inserts which were subcloned into the plasmid pTZ19R. The recombinant plasmids were transformed into the Escherichia coli strain, DHsa. Detailed restriction maps were produced of the 5.7Kb and 3.1Kb newt DNA inserts in the plasmids, designated pTNAl and pTNA2. The short «1.3 Kb) length of the actin homologous sequence in pTNA2 indicated that it was possibly a reverse transcript pseudogene. Problems associated with molecular cloning of DNA sequences from N. viridescens are discussed with respect to the large genome size and abundant highly repetitive DNA sequences.

Isolation of DNA sequences with homology to a degenerate FaRP oligonucleotide from the crayfish Procambarus clarkii

The neuropeptide Th1RFamide with the sequence Phe-Met-Arg-Phe-amide was originally isolated in the clam Macrocallista nimbosa (price and Greenberg, 1977). Since its discovery, a large family ofFl\1RFamide-related peptides termed FaRPs have been found to be present in all major animal phyla with functions ranging from modulation of neuronal activity to alteration of muscular contractions. However, little is known about the genetics encoding these peptides, especially in invertebrates. As FaRP-encoding genes have yet to be investigated in the invertebrate Malacostracean subphylum, the isolation and characterization ofFaRP-encoding DNA and mRNA was pursued in this project. The immediate aims of this thesis were: (1) to amplify mRNA sequences of Procambarus clarkii using a degenerate oligonucleotide primer deduced from the common amino acid sequence ofisolated Procambarus FaRPS, (2) to determine if these amplification products encode FaRP gene sequences, and (3) to create a selective cDNA library of sequences recognized by the degenerate oligonucleotide primer. The polymerase chain reaction - rapid amplification of cDNA ends (PCR-RACE) is a procedure in which a single gene-specific primer is used in conjunction with a generalized 3' or 5' primer to amplify copies ofthe region between a single point in the transcript and the 3' or 5' end of cDNA of interest (Frohman et aI., 1988). PCRRACE reactions were optimized with respect to primers used, buffer composition, cycle number, nature ofgenetic substrate to be amplified, annealing, extension and denaturation temperatures and times, and use of reamplification procedures. Amplification products were cloned into plasmid vectors and recombinant products were isolated, as were the recombinant plaques formed in the selective cDNA library. Labeled amplification products were hybridized to recombinant bacteriophage to determine ligated amplification product presence. When sequenced, the five isolated PCR-RACE amplification products were determined not to possess FaRP-encoding sequences. The 200bp, 450bp, and 1500bp sequences showed homology to the Caenorhabditis elegans cosmid K09A11, which encodes for cytochrome P450; transfer-RNA; transposase; and tRNA-Tyr, while the 500bp and 750bp sequences showed homology with the complete genome of the Vaccinia virus. Under the employed amplification conditions the degenerate oligonucleotide primer was observed to bind to and to amplify sequences with either 9 or 10bp of 17bp identity. The selective cDNA library was obselVed to be of extremely low titre. When library titre was increased, white. plaques were isolated. Amplification analysis of eight isolated Agt11 sequences from these plaques indicated an absence of an insertion sequence. The degenerate 17 base oligonucleotide primer synthesized from the common amino acid sequence ofisolated Procambarus FaRPs was thus determined to be non-specific in its binding under the conditions required for its use, and to be insufficient for the isolation and identification ofFaRP-encoding sequences. A more specific primer oflonger sequence, lower degeneracy, and higher melting temperature (TJ is recommended for further investigation into the FaRP-encoding genes of Procambarlls clarkii.

Oath of Allegiance to King George III

Transcript (original spelling and grammar retained): We the Commissioned Officers belonging to the Second Regiment of Lincoln Militia - Do sincerely promise and swear that we will be faithful and bear true allegiance to his Majesty King George, and heirs will defend to the utmost of our power against all traitorous conspiracies and attempts whatsoever which shall be made against his Person, Crown or Dignity; and we will do our utmost endeavours to disclose and make them known to his Majesty, His Heirs and successors, all treasons and traitorous conspiracies and attempts, which we shall know to be against Him or them. So help us God. Thomas Clark - Lt. Col. David Secord - Major John Crysler - Capt James Macklin - Capt John [Ross] - [Captain] [Abraham] Bowman - Lieut Gilbert McMicking - Quartmaster John [Misiner] - Ensign Robert Campbell - Capt John [Couke] - Ensign Nicholas Smith - Lieut I certify that the officers who have here [subscribed] the oath took it before me at Chippawa 4 Sept 1812 Thomas Dickson JP

Isolation and analysis of a corprinus cinereus DNA fragment showing homology to a fimbrial cDNA from ustilago violacea

Surface proteinaceous fibrils, termed fimbriae, were first identified on gram negative bacteria in the 1940s. Fungal fimbriae, discovered some 25 years later, are found on members of all fungal classes. In the present study, polyclonal antiserum raised against the fimbrial proteins of U. vio/acea were used in order to identify antigenically related proteins from Coprinus cinereus and Schizophy//um commune. Two polypeptides with molecular masses of 37 and 39 kDa from C. cinereus were observed and confirm earlier results. A single previously unidentified 50 kDa polypeptide in S. commune crossreacted with the antiserum. The 50 kDa protein was found to consist of 3 isoforms with isoelectric points ranging from 5.6 to 5.8. A fimbrial cDNA derived from U. vio/acea was used to identify DNA restriction fragments from C. cinereus and S. commune showing homology to the fimbrial transcript of U. vio/acea. Heterologous hybridization with this cDNA was used in order to screen a C. cinereus genomic DNA library. A single clone, A2-3A, with a 14 kbp insert showed strong homology to the pfim3-1 cDNA. The region of homology, a 700 bp Xba I fragment, was subcloned into pUG19. This plasmid was refered to as pXX8. DNA sequence determinations of pXX8 and adjacent fragments from A2-3A suggested that the cloned DNA was a portion of the rONA repeat encoding the small subunit rRNA. DNA sequence analysis of pfim3-1 yielded an incomplete open reading frame. The predicted amino acid sequence codes for a 206 amino acid, 22 kDa polypeptide which contains a domain similar to a transmembrane domain from rat leukocyte antigen, GDS3. As well, an untranslated 576 nucleotide domain showed 81 % homology to pXX8 and 830/0 homology to the 188 rRNA sequence of Ustilago maydis. This sequence was found adjacent to a region of adenine-thymine base pairs presumed to represent the polyadenylation sequence of the fimbrial transcript. The size and extent of homology is sufficient to account for the hybridization of pfim3-1 to rDNA. It is suggested that this domain represents a completely novel regulatory domain within eukaryotes that may enable the observed rapid regeneration of fimbriae in U. violacea.

Letter - American soldier's account of battles at Chippewa, Lundy's Lane and Fort Erie, 1814

Transcript (original grammar and spelling retained): My dear wife I take this time to inform you that I am well hoping that these few lines will Reach you and find you the same I shall in form you of all our Battles that we have had sence I left home we crossed in to Canada the 2 day of July and took fort Erie on the 3 day of July without loss of one man. We then marched down to Chipway eighteen miles below the Fort Erie we got there on the forth day and had our first battle on the 5 day our loss was not jistly known But the inemy loss was double to ours. The 6 day we started with the 2 Brigade to make a bridge a crost the crick two miles a bove the fort in Building the Bridge the inemy Brought up their Canon and playd upon us with their artiliery a bout two hours We drove them from the fort our loss was none the inemy loss was nineteen ciled dead on the ground we then marched to Queenston when we got thare our inemy had fledfrom the fort we then remained thair to Queenston ten days then we marched down to Fort George But that caurdly Chaney did not a rive with the fleet so we had to return back to Queenston thare was a bout six hundred militia formed on the heights of land thay fired up on us from their pickets and retreated to the mane body our flankers ciled and wounded and took about twenty before they got to the Maine body we then marched up the hill they gave us two firs but did not damage and then retreated from the field we stayed there one knight and then marched to Chipway and stayed there one night and the next day just as the sun set the first Brigade marched up in order to give them Battle a bout two miles from the Crick and began the Battle the 2 Brigade has to March up to the Niagara path and ingaged them we charged up on their artlery and took all their Canon Miller commanded the four companys that charged....the battles lasted three hours and forty minutes our loss was about 8 hundred cild and wounded our inemies loss was a bout fourteen hundred cild and wounded the next morning we Marched up in order to give them Battle a gin but thay was afraid to ingage us we then marched to Fort Erie and went to fortiffing and made a strong place the inemy folered us up and Began to cananade and held it fifty three days thay a tacked the fort the fifteenth of august thay atacked a bout one hiour be fore day Light we saw them and Blue up our maggerzean & two hundred of our inemy our loss wasa bout forty cild and wounded and our inemy loss was a bout one thousand on the 7 Day of September we atacked them and took their batteries and Broke all their canon and drove them from the field our loss was a Bout two hundred cild and wounded our inemy loss was a Bout 8 hundred cild and wounded...we crossed in to Canada with five thousand and came out with fifteen hundred we then Marched to Sackett’s harbor....am well and harty for the present....a bout comming home it uncarting for there is not any....given this winter as yet But I shall try to Come home if I Can But if I Cant I want you should take good car of the Phiddness[?] I have not Received any Money as yet But soon as I do receive some I send some home. I want you should write to me as soon as you receive this and and how Much Stock you wintor I Received your Letter with Great pleasure I feel uneasy a bout you I am a frade that you are sick or dead this is from your husband Chase Clough

Manuscript of James Cummings- August 28, 1860

Transcript (spelling and grammar retained): Chippawa [Chippewa] 28th August 1860 My Dear Sir I duly received your very kind letter of the 24th [June] asking me to communicate such facts of general interest connected with my career during the War with the United States. I have no objection to afford you such information as came under my own observation; nevertheless I do so, with the understanding, I have no desire to be my own trumpeter. With respect to your circular wherein you state you have been for several years collecting materials for a History of the late War between the United States & Great Britain, for which you are now gathering further materials to add to your collection, concerning the Second War for Independence. I am rather at a loss to know, what is meant by the second war; If you allude to the petty Rebellion, it could not be called a War, Those that caused the outbreak were very soon put down, by the Loyal people of the Province without the aid of Regular Troops being satisfied with the Independence they enjoyed. With respect to the several questions names in your circular: To the 1st I would say, this locality is made memorable by the battle of Chippawa [Chippewa] which took place about a mile above the village on the ground I pointed out to you, when I had the pleasure of seeing you a few days ago, with Mr Porter of the Niagara Falls, of which I believe you took sketches at the time. 2nd I have no historical documents of any value; so many years having gone past, the most of my old papers have either been lost or destroyed, I however came across two letters, one dated Queenston 9th July 1812 from Lt. Col. Nicholl Quarter Master General of Militia, the other from Lt. Col Myers Deputy Quarter Master General of the Regular Army date Fort George 23rd same month, directed to me in the hand writing of each of those officers as Deputy Quarter Master General of Militia, which letters I shall be obliged you would return at as early a day possible, as I wish to place them with tome others in the case, I have had made to hold the cocked hat & feather I wore during that eventful period, which I am sorry I did not exhibit when you was at my house; with reference to it I now enclose a letter from Lt. Col. Clark, residing at Port Dalhousie he was Captain & Adjutant of Militia in the War of 1812__ I send the letter in proof of the cock’d hat it is a lengthy one, but you may find time to turn over it, as I shall also place it in the hat case__ 3rd Where are [but] [for] traditionary [sic] witnesses residing in this vicinity – Col Clark above named Mr Merritt of St. Catharines, & Mr Kerby of Brantford are the only ones I now recollect, who could offord [sic] you any statistical information. 4th I have no pictorial sketches of any Military Movements or fortifications. As regards my own career, which you appear [ ? ] of knowing__ I was first a Lieutenant in a volunteer flank company stationed on the river side opposite [Navy] Island not far from the battle ground of Chippawa [Chippewa], I got promotion as Lieutenant of Cavalry before I got my Cavalry dress completed in three days more, I was called by General Brock to Fort George, was appointed Deputy Quarter Master General of Militia with the rank of Captain s the accompanying letters will show. I was at the battle of Stony Creek, several skirmishes at the Cross Roads, when the American army [ ? ] Fort George, at the taking of Col. Boerstler at the Beaver Dam, & had the honor of receiving Colonel Chapens sword at the surrender, who commanded a company of volunteer Horse Men was at the taking of 15 regulars & two officers at Fort Schlosser—was with Col. Bishop at the taking of Black Rock, near him when he fell, three men of the 8th Reg. more killed in the Boat I was in – I was at Chippawa battle, and the last, not the least in Lundy’s lane battle, which the Americans call the battle of Bridge [Waters]. I had forgot; there was another small affair at Corks Mill where I was. I could write a little history of events, but have not the time to do so. If what I have stated will be of any service for the purpose you require I shall feel happy. The history of the late War was published at Toronto in the Anglo American Magazine. Did you ever see it, I have the Books, there were however several errors which came under my notice, which I could have corrected. If my time would permit I could give you a more detailed statement of events. I trust however you may succeed with your publication , and I shall be most happy to hear from you at all times—I related many little occurances verbally to you when here, which I thought not necessary to repeat again as you would have a perfect recollection of them. Be pleased to return the letters for the purpose I require them. I am My Dear Sir Your respectful friend James Cummings

Letter from Lt.Col. Chewett to Ensign Kuck- 6 Sept 1814

Transcript (spelling and grammar retained): York 6th Sepber [September] 1814 Sir I will thank your for the Rolls and Returns of the Pickering Company due on the 1st instance that I may Make up mine to be returned to the Adjutant General – I have the honor to be Sir Your Most Obed [Obedient] W Chewett Lt Col 3rd Reg Yk [York] Militia

Letter From Sir Isaac Brock to James FitzGibbon - July 29, 1812

A two page letter written by Sir Isaac Brock in York, Upper Canada to James FitzGibbon on July 29, 1812. The name of the recipient is not included but according to Mary Agnes FitzGibbon, one can find a transcript of the letter in her "A Veteran of 1812", page 60.[1812], 29 July: Major-General Isaac Brock, York, to James FitzGibbon. I lament that you should have been so long impressed with the idea that I possessed the means of being serviceable to you. I had scarcely heard of Mr. Johnson having declined a Company in the Glengarry (which would have given me the nomination) but I received account of his being reinstated. I consequently thought no more of the business thinking that officer was enjoying the fruits of his good fortune. I know not positively whether Mr. Johnson is reinstated, but being under obligations to promote his views, I cannot possibly interfere to his prejudice. I rather wonder you did not hear that Lieut Lamont had long ago my promise of nominating him to the Company provided it became vacant, which of course would have precluded my application in your behalf. Altho you must be sensible of the impossibility of my taking any step to forward your views in the present case, yet be assured I shall always feel happy in any opportunity that may offer to do your service. To a person unaccustomed to my writing I scarcely would hazard sending this scrawl. I am, Dear Sir, Yours faithfully, Isaac Brock I should like to be among the 49th at this moment. I am satisfied they will support and even add to their former fame. They have my very best wishes. The 41st are behaving nobly at Amherstburg.

Letter from David Todd to Colonel Proctor, Commanding Officer of the 41st Regiment- 9 Feb 1812

Transcript (spelling and grammar retained): “Col Proctor Sir I hope your goodness will excuse the Liberty I have taken of Enclosing a Letter for my nephew Mr. Hailes to your care, and begging the favor of you to forward it to him, - not knowing myself at what Post he is – With Great Respect I am Sir Your Most Obed Serv David Todd”

John Adams Smith letter, Jan. 14, 1813

Transcript: New York Jan. 14. 1813. My dear uncle, I had nearly concluded to have made my [jaunt?] earlier than I had mentioned in my last letter to you, for Swartout had intended to have written by me to Father & yourself; having [Louisa?] concluded not so soon to depart he sends me his letters which I forward in mail, I shall hope to have letters from home soon, it is a great pleasure to hear from any of you. I said last week in Hollands letter mentioning [bladeworth?] account, I have explained to him. It is a mistake about me agreeing to pay Mr. [Dening?] [their?] account. Caroline mentions to me your jaunt to [Sophia?], but I regret our friend there should be unpleasantly situated. I have not yet seen Col. Willett but will see him as mentioned. I have neglected to go there for some time which has been remiss in me. We have nothing new I believe unless it be that Armstrong & Jones of Philadelphia have been nominated as Secretarys of War & the Navy & [cer?] this in all probability may be appointed. There may be room for speculation, but perhaps it is not probable that Canada will be ceded to American Valor in the year 1813. There is a Bill before congress by which it may be made penal to enter on board American vessels, either British subjects or naturalized Americans, which it is presoomed by many will [spon?] the accommodation of Peace – may be yes may be no – the proof of the pudding is in the eating of it. I imagine pacification is not so near at hand although it is much to be desired. Whatever might be for the honor & prosperity of the Country I would strenuously advocate, aloof from partial & party considerations. We have not a word as yet what the Council at Albany may be thinking of as yet. I suppose these things will come in time, perhaps untimely to many. I desire for the present not to be found among the untimely. I am your sincerely, with love to all. John Adams Smith

R.Lawson letter

Transcript (original spelling and grammar retained): Chatham 17th Sept. 1812. Sir, I have just received a letter from Cap’n Clay informing me that boats will be ready at the Communication Bridge tomorrow morning by day break to return that part of the 9th Reg’t which are to embark on board the Success[?] and Melabar Troop Ships[?]—and that they will likewise be ready to receive [?] on Saturday next at the same time, the division intended for the Diomedes [?]. I have therefore given the Regiment orders accordingly –and will give every assistance respecting the heavy baggage required for its safety, until some arrangement is made for its conveyance from here—the Barrack Master having no vacant store for the purpose. I have the honor to be Sir your obed’t Ser’t R. Lawson M. Gen’l. Ass Qr Master Gen’l Canterbury. Chatham 17 Sept. 1812. M. …Lawson. That boats will be in readiness at day break on the 18th & 19th Ins. For convy. the 9th Reg &…..[?] to Baggage of that Corps …..

Letter written by John Bentley, Sept. 26, 1813.

Transcript [original spelling and grammar retained]: Sacketts Harbour 26th Sept. 1813. Dear wife. From this you are informed that I am in health at this Date. When I last wrote to you I some expected to go upon and expedition but to us unknown we sat out and went up the Lake 3 Days Landed at Oswego The British Fleet appeared off, and we returned and saw our fleet upon our return, I think that our Regt. going was only a maneuver to get the Fleet out that our Fleet might come a threat[?] of them We expect to embark immediately into Canada as preparations are making to convey us over to them we are anxious to commence an action with them. Troops are daily coming in to the Harbour to take the Stand in our absence, we shall not be here 3 days before we try their[?] Powder. they are daily defecting[?] to us from Canada very fearful of the consequence of our Resisting[?] of them…our Fleet is now out and has been for 6 or 7 days. The Lake Erie fleet has done great things. I hope ours will be as successful. I think that our Generals are waiting to hear from them as every thing is ready[?] of to embark various opinions reflecting were we shall attack them some say at Kingston others at Montreal and others at Prescot—Mr. William Butler and D[?]…are well and in Spirits, Sergt Daniel White is very hearty for him, M. Samuel C[?] is well and in good health Benjm Thompson is well Charles Bryant is well [?] is well Eben[?] Smith is very much plagued with the Rheumatik Disease[?] he……… his limbs very often for being Crippled[?] he is at the Hospital I often visit it to see the sick Jacob Barnes is at the Hospital but recovering fast been very sick. Luther Gregory is at the Hospital and on the recovery, Sergt. L[?] & Smith are well, Henry ………[?] is well, very healthy have not more die here than 3…[?]to the best of my knowledge. I will make a few remarks upon the place it abounds in Lime Rock more than Thomaston and not every person to my knowledge burn it, and in the whole Town not but one Pump that supplies the Towns People and Soldiers and a ………[?] of such a Lake of water the Lake water is good for drinking but the water near the Shore is exposed to all kind of filth being thrown into it. The officers with whom I have been with have used me kindly and I get quietly by them. The Lieut. Downer who recruits at Thomaston tell me he has thots[?] of Leaving the army if so I must say I am greatly sorry as he was my ……[?] friend although he Left us and went in a northern Company it is a Company……………[?]worthy an officer as he proves to be, I cannot get any higher than a Sergeant or Quarter Master Sergeant which I may have without any friends at Thomaston assisting me. I am a Sergeant and Sergeant Daniel White is expecting[?] to be a Quarter Master Sergeant, and a number of his friends from Thomaston have went to their Major for him in the 9th Regt Major….[?] and he expects to obtain a Commission as I ……[?] expect to be promoted and it died away he will have the Laugh upon me, I wish that My Friend Dawes would[?] put the question to Col. Foot? to write to our Col. E.W. Ripley if he has …[?]in the …[?]taken by him and others of my …[?] friends[?]. I expect to come home this winter without fail. I remain your Loving Husband till Death. John Bentley for Betsey Bentley Thomaston P.S. The next Letter will be ……[?]to Mr. Dawes[?] and shall write as soon as our Fleet arrives or if we are ordered off tomorrow shall write before I leave this Place. I have understood that many letters have been ……[?]to me. I have received only 2 from Mary, one from Mr. Dawes, one from William Thompson and have answered them please to write……[?]to S. Harbour.

Daniel D. Tompkins letter to General Dearborn, June 28, 1812.

Transcript [original spelling and grammar retained]: Albany June 28 1812 Sir Your letter of the 23d has been received. I had anticipated your request by ordering the detachment from Washington, Essex, Clinton and Franklin Counties into service and have fixed the days and places of their Rendezvous. Upon application to the quarter master General I find there are but 139 tents & 60 camp kettles at this place & even those I take by a kind of stealth. The Deputy Quarter Master General declines giving an order for their delivery until he shall have a written order from the Quarter Master General, and the latter is willing I Shall take them but will not give the Deputy a written order for that purpose. Under Such circumstances I shall avail myself of the rule of Possession and by virtue of the Eleven Points of Law Send them tomorrow morning without a written order from anyone. You may remember that when you were Secretary of the war department I invited you to forward and deposit in our Frontier Arsenals, arms ammunition and camp equipage free of expense to be ready in case of war and the same invitation to the war department has been repeated some time, Since The United States have now from 5 to 600 regular troops at Plattsburgh, Rome, Canandaigua & c, where those arsenals are, and yet those recruits are now and must be for weeks to come unarmed and unequipped in every respect although within musket Shot of arsenals. The recruits at Plattsburgh are within 50 miles of two tribes of Canadian Indians. In case of an attack upon the Frontiers that portion of the United States army would be as inefficient and unable to defend the inhabitants or themselves even. The Militia Detachments on the Western Frontiers received the news of war with [cherafulness] and determined courage, and I am happy to find they are united late brothers, highly improved in Discipline & ready to devote themselves to any Service or danger which the good of the Country may require. But they are in barracks from which they cannot move a days march for the want of tents and other equipage, and they are in Separate and Independent Detachments without a General Officer to command them or combine their exertions for the accomplishment of any desirable and important object. The only officer of the United States here who can do anything is the Quarter Master General and he has not a tent Camp Kettle or Knapsack in this arsenal except what I have concluded to send off tomorrow morning as above mentioned to furnish men As to Cannon Muskets and Ammunition. I can find now one here who will exercise any authority over them or deliver a Single article upon my requisition. Neither can I find any Officer of the army who feel himself authorized to exercise any authority or do any act which will aid me in the all important object of protecting the Inhabitants of an extended Frontier exposed to the Cruelties of Savages and the depredations of the enemy. If I must rely upon the Militia Solely for Such protection I entreat you to give orders to your Officers to furnish upon my orders, for the use of the Militia Detachments , all needful weapons and articles with which the United States are Supplied and of which we are destitute . You may rely upon all the assistance which my talents, influence or authority can furnish in the active prosecution of the first & necessary way which has been declared by the Constituted Authorities of our beloved Country. I am, Sir, respectfully Your ob. Servt. Daniel D. Tompkins

Grant of Permission for lease of military lands from Sir Robert Prescott to Thomas Clark, 1798-1801.

Transcript (original spelling and grammar retained): By His Excellency Robert Prescott Esquire, Captain General and Governor in Chief in and over His Majestys Provinces of Upper and Lower Canada, General and Commander in Chief of all His Majesty’s forces in the Provinces of Upper and Lower Canada, Nova Scotia, New Brunswick and their several Dependencies and in the Island of Newfoundland &ca. &ca. &ca. I do hereby authorise and permit Thomas Clark of Queenstown in the County of Lincoln in the Province of Upper Canada merchant to take possession of all that Lot, piece and parcel of Land (being part of the land reserved by his Majesty for Military purposes) situate, lying and being at Queenstown in the Township of Newark, in the Home District in the said Province of Upper Canada, bounded and abutted as follows, that is to say beginning at the Distance of two Chains and ninety links from the South East End of his Majesty’s Store House, the said distance being measured along the Bank up Stream, thence South thirty nine degrees and an half West one Chain and fifty links thence south fifty degrees and an half East one Chain and thirty links thence North thirty nine degrees and an half East to the Edge of the Bank and from thence along the Bank to the place of beginning, containing thirty one perches and one hundred and twenty five square links and to occupy and hold the said Lot, piece and parcel of Land during pleasure subject nevertheless to the provisoes and Conditions herein after contained, that is to say. First on condition that it shall and may be lawful to and for His Majesty his Heirs and Successors and to and for the Commander in Chief of His Majesty Forces for the time being and to and for the Officer commanding his Majesty’s Forces in Upper Canada for the time being and to and for either of them to determine and make void this present permission to occupy during pleasure the said Lot, Piece or Parcel of Land above described at any time hereafter whenever he or they shall see fit so to do without any compensation or indemnification to the said Thomas Clark or any other Person or Persons whosoever for any Loss Injury or Damage which he the said Thomas Clark or any other Person or Persons whosoever may thereby sustain. Secondly on this further Condition that it shall and may be lawful to and for His Majesty his Heirs and Successors and to and for his and their Officers, Soldiers and Servants at any time hereafter by order of the Commander in Chief of His Majesty’s Forces for the time being or by order of the Officer commanding his Majesty’s forces in Upper Canada for the time being or by order of the Officer of His Majesty’s Corps of Royal Engineers commanding in the said Province of Upper Canada for the time being to enter upon the said Lot Piece and parcel of Land which the said Thomas Clark is hereby permitted to occupy during pleasure or upon any part thereof and to take down and from the said Lot piece and parcel of Land or from any part thereof to remove any dwelling House Store or other Buildings on the said Lot, piece or Parcel of Land or any part thereof erected and to remove any goods or Chattels on the said Lot piece and parcel of Land or on any part thereof or on any such dwelling House Store or other building found or being and that His Majesty his Heirs and Successors or any other Person or Persons whosoever shall not be liable or responsible to the said Thomas Clark or to any other Person or Persons whosoever for any Loss, Injury or Damage which he or they shall or may in such case sustain. Thirdly on this further Condition that the said Thomas Clark shall not erect on the said Lot Piece or Parcel of Land which the said Thomas Clark is hereby permitted to occupy during pleasure or upon any part thereof at any time or times hereafter any dwelling House store or other Building whatsoever of Stone or brick or of any other materials wood only exccepted and that if any dwelling House or Store or other building of Stone or brick or of any other materials except wood shall at any time be erected on the said Lot, piece or parcel of Land or upon any part thereof, then and in such case, this present permission and every Clause and Article thereof shall from thenceforth cease and determine and be absolutely and entirely null and void. And lastly on this further Condition that the said Thomas Clark or any other Person whosoever shall not assign this permission to occupy the said Lot, Piece or Parcel of Land above described to any Person of Persons whosoever, and if any such assignment shall be made by the said Thomas Clark or by any other Person in his right, or on his behalf, that then and in such case such assignment and this permission to occupy during pleasure the said Lot piece and parcel of Land above described, and every Clause and Article thereof shall from thenceforth cease and determine and be absolutely and entirely null and void. Given under my hand at the Castle of St. Lewis in the City of Quebec in the Province of Lower Canada this Ninth day of July in the year of our Lord one thousand seven hundred and ninety eight and in the thirty eighth year of His Majesty’s Reign. [Signed here by Robert Prescott] By Order of the Commander in Chief [Signed here by James Green (Illegible signature)] I the said Thomas Clark above named do hereby accept the above written Permission to occupy during pleasure the said Lot piece and parcel of Land above described upon and subject to the several Provisioes and Conditions above written and each and every of them severally and respectively. In witness whereof I have hereunto set my hand the Sixteenth day of August in the year of our Lord one thousand seven hundred and ninety eight and in the thirty eighth year of His Majesty’s Reign. [Signed here by Thomas Clark] Signed in the presence of [Illegible signature – looks like J. M Donell Lt. Col.] [Illegible – looks like 2d. Battn R. C. Sm?] [Signed here by C. Anderson] Whereas Inconveniences did arise from the peculiar situation of the Ground contiguous to the above described Lot of Land and the occupation thereof, if estimated by the above Limits would prove highly disadvantageous to Mr. Thomas Clark be it known that in consideration thereof we do permit the above Lot to extend one half Chain more in length up stream so as to comprehend the space allowed for the Road between Lots Two + Three, and we do hereby appropriate the said additional space wholly to the use of the said Thomas Clark. In witness whereof we have hereunto subscribed this Thirteenth Day of October in the Year of our Lord one thousand Eight Hundred and one. [Signed here by J. M’Donell Lt. Col] 2d. Battn. R. C. [in?] Com of Fort George + Dependencies Robt. Pilkington Captain Royal Engineers

Defence of Agaricus bisporus against toxic secondary metabolites from Trichoderma aggressivum.

Trichoderma spp are effective competitors against other fungi because they are mycoparasitic and produce hydrolytic enzymes and secondary metabolites that inhibit the growth of their competitors. Inhibitory compounds produced by Trichoderma aggressivum, the causative agent of green mold disease, are more toxic to the hybrid off-white strains of Agaricus bisporus than the commercial brown strains, consistent with the commercial brown strain’s increased resistance to the disease. This project looked at the response of hybrid off-white and commercial brown strains of A. bisporus to the presence of T. aggressivum metabolites with regard to three A. bisporus genes: laccase 1, laccase 2, and manganese peroxidase. The addition of T. aggressivum toxic metabolites had no significant effect on MnP or lcc1 transcript abundance. Alternatively, laccase 2 appears to be involved in resistance to T. aggressivum because the presence of T. aggressivum metabolites results in higher lcc2 transcript abundance and laccase activity, especially in the commercial brown strain. The difference in laccase expression and activity between A. bisporus strains was not a result of regulatory or coding sequence differences. Alteration of laccase transcription by RNAi resulted in transformants with variable levels of laccase transcript abundance. Transformants with a low number of lcc transcripts were very sensitive to T. aggressivum toxins, while those with a high number of lcc transcripts had increased resistance. These results indicated that laccase activity, in particular that encoded by lcc2, serves as a defense response of A. bisporus to T. aggressivum toxins and contributes to green mold disease resistance in commercial brown strains.