Restriction fragment length polymorphism-coupled domain-directed differential display: a highly efficient technique for expression analysis of multigene families.

In this paper, a reverse-transcriptase PCR-based protocol suitable for efficient expression analysis of multigene families is presented. The method combines restriction fragment length polymorphism (RFLP) technology with a gene family-specific version of mRNA differential display and hence is called "RFLP-coupled domain-directed differential display. "With this method, expression of all members of a multigene family at many different developmental stages, in diverse tissues and even in different organisms, can be displayed on one gel. Moreover, bands of interest, representing gene family members, are directly accessible to sequence analysis, without the need for subcloning. The method thus enables a detailed, high-resolution expression analysis of known gene family members as well as the identification and characterization of new ones. Here the technique was used to analyze differential expression of MADS-box genes in male and female inflorescences of maize (Zea mays ssp. mays). Six different MADS-box genes could be identified, being either specifically expressed in the female sex or preferentially expressed in male or female inflorescences, respectively. Other possible applications of the method are discussed.

Cytotoxic T-lymphocyte clones from different patients display limited T-cell-receptor variable-region gene usage in HLA-A2-restricted recognition of the melanoma antigen Melan-A/MART-1.

To determine whether T-cell-receptor (TCR) usage by T cells recognizing a defined human tumor antigen in the context of the same HLA molecule is conserved, we analyzed the TCR diversity of autologous HLA-A2-restricted cytotoxic T-lymphocyte (CTL) clones derived from five patients with metastatic melanoma and specific for the common melanoma antigen Melan-A/MART-1. These clones were first identified among HLA-A2-restricted anti-melanoma CTL clones by their ability to specifically release tumor necrosis factor in response to HLA-A2.1+ COS-7 cells expressing this tumor antigen. A PCR with variable (V)-region gene subfamily-specific primers was performed on cDNA from each clone followed by DNA sequencing. TCRAV2S1 was the predominant alpha-chain V region, being transcribed in 6 out of 9 Melan-A/MART-1-specific CTL clones obtained from the five patients. beta-chain V-region usage was also restricted, with either TCRBV14 or TCRBV7 expressed by all but one clone. In addition, a conserved TCRAV2S1/TCRBV14 combination was expressed in four CTL clones from three patients. None of these V-region genes was found in a group of four HLA-A2-restricted CTL clones recognizing different antigens (e.g., tyrosinase) on the autologous tumor. TCR joining regions were heterogeneous, although conserved structural features were observed in the complementarity-determining region 3 sequences. These results indicate that a selective repertoire of TCR genes is used in anti-melanoma responses when the response is narrowed to major histocompatibility complex-restricted antigen-specific interactions.

Expansible Computable Aggregation Rules

The aggregation operator have been considered from a computable point of view. The important condition that the computation is friendly when portions of data are inserted o deleted to the list of values to aggregate is considered.

Staying in the Delaware Corporate Governance Lane: Fee Shifting Bylaws and a Legislative Reaffirmation of the Rules of the Road

The Delaware legislature has taken steps towards the adoption of amendments to the Delaware General Corporation Law (DGCL) that would prohibit fee shifting provisions in the articles and bylaws. The language in the legislative proposal, however, addresses fee shifting provisions only in the context of "internal corporate claims." Some have raised concerns that this language would allow for fee shifting provisions that applied to other types of actions, including at least some cases brought under the securities laws. This piece suggests that in fact the Delaware General Corporation Law already prohibits the adoption of bylaws and certificate provisions that apply to causes of action unrelated to internal corporate claims. As a result, there was no reason for the Delaware legislature to expressly bar fee shifting provisions in these types of actions.

Representing Clients in the Marijuana Industry: Navigating State and Federal Rules

As the number of states legalizing medicinal and recreational marijuana increases and marijuana emerges as a growing lawful industry, lawyers find themselves in an awkward position. In most states, lawyers who represent clients in the marijuana industry risk discipline for assisting clients in the commission of a (federal) crime. Even in jurisdictions like Colorado, where the rules of professional conduct have been amended to permit lawyers to assist clients who comply with marijuana state laws, lawyers who are admitted to practice in federal courts risk being disciplined by these tribunals for assisting clients in the commission of a crime pursuant to the courts’ local rules of conduct. This short article explores the thorny issue of navigating state and federal rules of professional conduct while representing clients in the marijuana industry.

Rules and orders respecting the Steward, Cook & Butler of Harvard College, [1686]

These regulations are a revision of the original 1650 rules that laid out the duties of the Steward, Butler, and Cook. In these updated regulations, the Rector and Tutor are now responsible for signing off on the Steward's accounts rather than the President and Fellows.

E. Committee to frame rules, directions, and statutes of the Boylston Professorship of Rhetoric and Oratory, 1804

This subseries consists of a paper notebook containing a handwritten draft of the report presented to the Harvard Corporation on April 30, 1804 by the Committee to frame Rules, Directions, and Statutes of the Boylston Professorship of Rhetoric and Oratory. The handwritten report provides a numbered list of rules related to the Boylston Professorship and is dated April 16, 1804. The report is followed by a certification signed May 1, 1804 from President Joseph Willard that he was unable to attend the meeting of the Corporation to discuss the professorship.

Draft of the Rules, Directions, and Statutes of the Boylston Professorship of Rhetoric and Oratory in Harvard College, [ca. 1804]

This original draft was probably written by Eliphalet Pearson (1752-1826) as a member of the committee charged with the task of establishing the rules, directions, and statutes for the Boylston Professorship by the Harvard Corporation and the Board of Overseers. This draft is heavily edited and contains many cross outs through the text.

College Book 7, 1750-1778

This volume contains a fair copy of minutes from Corporation meetings held from Sept. 17, 1750 through April 23, 1778. It begins with an alphabetical index and contains entries related to a wide range of topics, including the challenges of operating the Charlestown ferry (due to the river freezing, fear of smallpox, and other issues); increases in "pecuniary mulcts" (fines) for breaches of specific College laws; the establishment of the Dudleian lecture; the selection and financial support of missionaries to various Indian tribes; honorary degrees awarded to Benjamin Franklin and George Washington; gifts to the library as it was rebuilt in the wake of the fire of 1764 (many entries provide the title and author of books donated); the management of land and property belonging to Harvard; Treasurers' reports and other financial accounts; changes in the College laws; gifts to the College, ranging from two Egyptian mummies to a solar microscope; the construction of the First Parish Meeting House in Cambridge and the use of adjacent College property by parishoners; rules of endowed professorships; salaries and appointments; closures due to the threat of smallpox; rules governing Commons and the College Library; reports of various Visiting Committees; class schedules, according to subject; student disorders; the establishment of a designated museum space to display "Curiosities"; the effects of the Revolutionary War on Harvard, including repeated requests to the General Court after the war for compensation for damage to College buildings; the cost of various foods and changes in what was served at Commons; and the danger of the chapel's roof, built of too-heavy slate, falling in. Also of interest are minutes from a May 5, 1761 meeting, which note that the General Court voted to pay for Hollis Professor John Winthrop to travel to Newfoundland to observe the transit of Venus "over the Suns disc."

E. Committee to frame rules, directions, and statutes of the Boylston Professorship of Rhetoric and Oratory, 1804

This subseries consists of a paper notebook containing a handwritten draft of the report presented to the Harvard Corporation on April 30, 1804 by the Committee to frame Rules, Directions, and Statutes of the Boylston Professorship of Rhetoric and Oratory. The handwritten report provides a numbered list of rules related to the Boylston Professorship and is dated April 16, 1804. The report is followed by a certification signed May 1, 1804 from President Joseph Willard that he was unable to attend the meeting of the Corporation to discuss the professorship.

Rules and orders relating to a Professor of the Mathematicks, of Natural and Experimental Philosophy, in Harvard College, 18 January 1726

This folder contains a single document describing the "rules and orders" of the Hollis Professor of Mathematics and Natural Philosophy. The document begins by defining the subjects to be taught by the Hollis Professor including natural and experimental philosophy, elements of geometry, and the principles of astronomy and geography. It then outlines the number of public and private lectures to be given to students, how much extra time the professor should spend with students reviewing any difficulties they may encounter understanding class subject matter discussed, and stipulates that the professor's duties shall be restricted solely to his teaching activities and not involve him in any religious activities at the College or oblige him to teach any additional studies other than those specified for the Hollis Professor of Mathematics and Natural Philosophy. Furthermore, the rules establish the professor's salary at £80 per year and allow the professor to receive from students, except those students studying theology under the Hollis Professor of Divinity, an additional fee as determined by the Corporation and Board of Overseers, to supplement his income. Moreover, the rules assert that all professorship candidates selected by the Harvard Corporation must be approved by Thomas Hollis during his lifetime or by his executor after his death. Finally, the rules state that the Hollis professor take an oath to the civil government and declare himself a member of the Protestant reformed religion. This document is signed by Thomas Hollis and four witnesses, John Hollis, Joshua Hollis, Richard Solly, and John Williams.