Determining RNA solution structure by segmental isotopic labeling and NMR: application to Caenorhabditis elegans spliced leader RNA 1.

Recent developments in multidimensional heteronuclear NMR spectroscopy and large-scale synthesis of uniformly 13C- and 15N-labeled oligonucleotides have greatly improved the prospects for determination of the solution structure of RNA. However, there are circumstances in which it may be advantageous to label only a segment of the entire RNA chain. For example, in a larger RNA molecule the structural question of interest may reside in a localized domain. Labeling only the corresponding nucleotides simplifies the spectrum and resonance assignments because one can filter proton spectra for coupling to 13C and 15N. Another example is in resolving alternative secondary structure models that are indistinguishable in imino proton connectivities. Here we report a general method for enzymatic synthesis of quantities of segmentally labeled RNA molecules required for NMR spectroscopy. We use the method to distinguish definitively two competing secondary structure models for the 5' half of Caenorhabditis elegans spliced leader RNA by comparison of the two-dimensional [15N] 1H heteronuclear multiple quantum correlation spectrum of the uniformly labeled sample with that of a segmentally labeled sample. The method requires relatively small samples; solutions in the 200-300 microM concentration range, with a total of 30 nmol or approximately 40 micrograms of RNA in approximately 150 microliters, give strong NMR signals in a short accumulation time. The method can be adapted to label an internal segment of a larger RNA chain for study of localized structural problems. This definitive approach provides an alternative to the more common enzymatic and chemical footprinting methods for determination of RNA secondary structure.

Degradation of the cleaved leader peptide of thiolase by a peroxisomal proteinase.

A peroxisomal location for insulin-degrading enzyme (IDE) has been defined by confocal immunofluorescence microscopy of stably transfected CHO cells overexpressing IDE and digitonin-permeabilization studies in normal nontransfected fibroblasts. The functional significance of IDE in degrading cleaved leader peptides of peroxisomal proteins targeted by the type II motif was evaluated with a synthetic peptide corresponding to the type II leader peptide of prethiolase. The peptide effectively competed for degradation and cross-linking of the high-affinity substrate 125I-labeled insulin to IDE. Direct proteolysis of the leader peptide of prethiolase was confirmed by HPLC; degradation was inhibited by immunodepletion with an antibody to IDE. Phylogenetic analysis of proteinases related to IDE revealed sequence similarity to mitochondrial processing peptidases.

Schleiermacher's Doctrine of Biblical Authority: An Alternative to Content-Based/Supernaturalist and Function-Based Rationalist Models

Author: Kerry W. Holton Title: SCHLEIERMACHER’S DOCTRINE OF BIBLICAL AUTHORITY: AN ALTERNATIVE TO CONTENT-BASED/SUPERNATURALIST AND FUNCTION- BASED/RATIONALIST MODELS Advisor: Theodore M. Vial, Jr. Degree Date: August 2015 This dissertation examines Friedrich Schleiermacher’s understanding of biblical authority and argues that, as an alternative to strictly supernaturalistic and rationalistic models, his understanding allows the New Testament to speak authoritatively in Christian religion in an age of critical, historical awareness. After classifying Schleiermacher’s position in a typology of the doctrine of biblical authority, this dissertation explores his conception of divine revelation and inspiration vis-à-vis scripture. It demonstrates that although he did not believe there is warrant for the claim of a direct connection between divine revelation and scripture, or that scripture is the foundation of faith, he nonetheless asserted that the New Testament is authoritative. He asserted the normative authority of the New Testament on the basis that it is the first presentation of Christian faith. This dissertation examines Schleiermacher’s “canon within the canon,” as well as his denial that the Old Testament shares the same normative worth and inspiration of the New. Although this dissertation finds difficulty with some of Schleiermacher’s views regarding the Old Testament, it names two significant strengths of what is identified as his evangelical, content-based, and rationalist approach to biblical authority. First, it recognizes and values the co-presence and co-activity of the supernatural and the natural !ii in the production of the New Testament canon. This allows both scripture and the church to share religious authority. Second, it allows Christian faith and the historical-method to coexist, as it does not require people to contradict what they know to be the case about science, history, and philosophy. Thus, this dissertation asserts that Schleiermacher’s understanding of biblical authority is a robust one, since, for him, the authority of scripture does not lie in some property of the texts themselves that historians or unbelievers can take away.

Payment order for Moses Parsons on the behalf of his son, 1767 July 1

Handwritten order to John Sale to pay scholarship funds to Moses Parsons on behalf of his son, signed by Thomas Foxcroft, Charles Chauncey, Thomas Waite, and Daniel Marsh.

[Map of the Dead Sea before the Biblical destruction of Sodom, 1812 May 4]

A pen-and-ink and watercolor map of the Dead Sea noting the "Course of Jordan before the destruction of Sodom." The map accompanied a letter by Winthrop (HUG 1203.5 Box 1, Folder 13).

Moses Parker's Release to Hopkinston [sic] Trustees, 1716 December 21

Handwritten copy of a deed between Moses Parker and the Trustees for the land originally purchased by Parker, et al. from John Owosamog in 1700.

Notebook of Moses Appleton, 1794-1837 (inclusive)

Contains notes taken by Moses Appleton (1773-1849) on anatomy lectures delivered at Harvard by John Warren (1753-1815). Other lecture topics included midwifery and surgery. Also includes a transcript of an examination given by Warren to his students on anatomy and surgery, as well as exams given by Harvard Professor Benjamin Waterhouse (1754-1846) and Harvard Professor Aaron Dexter (1750-1829) on the theory and practice of physic, and chemistry, respectively. There are additionally patient case notes and transcriptions of notes and correspondence from physicians Appleton consulted, and a list of operations Appleton performed between 1796 and 1828, primarily repairing dislocated joints and fractured bones. Also includes obituaries of citizens of Waterville, Maine, from 1807 to 1837.

Commonplace book of Moses Appleton, 1791-1815 (inclusive)

Volume containing medicinal recipes, medical notes, poetry, and obituaries written by Dr. Moses Appleton (1773-1849). Many of the recipes were copied from medical texts or other publications. His "cure for the dropsy," taken from the New York Herald, contained stale cider, parsley, horseradish, oxymel squills (sea onion in honey), and juniper berries. For diarrhea, he prescribed a blackberry syrup. Several entries indicate Appleton practiced Thomsonian medicine, an alternative system based on use of botanicals. The medical notes include an account of his treatment of a man with smallpox in 1815, and entries on patients he inoculated with cowpox matter. Another entry dated in 1796 provides instructions from the Massachusetts Humane Society for "treatment to be used with persons apparently dead from drowning," which included blowing tobacco smoke in the victim's lungs and applying warm blankets for several hours. Appleton adds a note questioning whether or not the lungs also should be "often artificially inflated." There is additionally a history of prominent physicians dating from ancient Greece.