Molecular cloning and sequence analysis of the complestatin biosynthetic gene cluster

Streptomyces lavendulae produces complestatin, a cyclic peptide natural product that antagonizes pharmacologically relevant protein–protein interactions including formation of the C4b,2b complex in the complement cascade and gp120-CD4 binding in the HIV life cycle. Complestatin, a member of the vancomycin group of natural products, consists of an α-ketoacyl hexapeptide backbone modified by oxidative phenolic couplings and halogenations. The entire complestatin biosynthetic and regulatory gene cluster spanning ca. 50 kb was cloned and sequenced. It consisted of 16 ORFs, encoding proteins homologous to nonribosomal peptide synthetases, cytochrome P450-related oxidases, ferredoxins, nonheme halogenases, four enzymes involved in 4-hydroxyphenylglycine (Hpg) biosynthesis, transcriptional regulators, and ABC transporters. The nonribosomal peptide synthetase consisted of a priming module, six extending modules, and a terminal thioesterase; their arrangement and domain content was entirely consistent with functions required for the biosynthesis of a heptapeptide or α-ketoacyl hexapeptide backbone. Two oxidase genes were proposed to be responsible for the construction of the unique aryl-ether-aryl-aryl linkage on the linear heptapeptide intermediate. Hpg, 3,5-dichloro-Hpg, and 3,5-dichloro-hydroxybenzoylformate are unusual building blocks that repesent five of the seven requisite monomers in the complestatin peptide. Heterologous expression and biochemical analysis of 4-hydroxyphenylglycine transaminon confirmed its role as an aminotransferase responsible for formation of all three precursors. The close similarity but functional divergence between complestatin and chloroeremomycin biosynthetic genes also presents a unique opportunity for the construction of hybrid vancomycin-type antibiotics.

Gain of chromosome 3q defines the transition from severe dysplasia to invasive carcinoma of the uterine cervix.

We have chosen tumors of the uterine cervix as a model system to identify chromosomal aberrations that occur during carcinogenesis. A phenotype/genotype correlation was established in defined regions of archived, formalin-fixed, and hematoxylin/eosin-stained tissue sections that were dissected from normal cervical epithelium (n = 3), from mild (n = 4), moderate (n = 6), and severe dysplasias/carcinomas in situ (CIS) (n = 13), and from invasive carcinomas (n = 10) and investigated by comparative genomic hybridization. The same tissues were analyzed for DNA ploidy, proliferative activity, and the presence of human papillomavirus (HPV) sequences. The results show that an increase in proliferative activity and tetraploidization had occurred already in mildly dysplastic lesions. No recurrent chromosomal aberrations were observed in DNA extracted from normal epithelium or from mild and moderate dysplasias, indicating that the tetraploidization precedes the loss or gain of specific chromosomes. A gain of chromosome 3q became visible in one of the severe dysplasias/CIS. Notably, chromosome 3q was overrepresented in 90% of the carcinomas and was also found to have undergone a high-level copy-number increase (amplification). We therefore conclude that the gain of chromosome 3q that occurs in HPV16-infected, aneuploid cells represents a pivotal genetic aberration at the transition from severe dysplasia/CIS to invasive cervical carcinoma.

Activation of a 15-kDa endonuclease in hypoxia/reoxygenation injury without morphologic features of apoptosis.

Hypoxia/reoxygenation is an important cause of tissue injury in a variety of organs and is classically considered to be a necrotic form of cell death. We examined the role of endonuclease activation, considered a characteristic feature of apoptosis, in hypoxia/reoxygenation injury. We demonstrate that subjecting rat renal proximal tubules to hypoxia/reoxygenation results in DNA strand breaks and DNA fragmentation (both by an in situ technique and by agarose gel electrophoresis), which precedes cell death. Hypoxia/reoxygenation resulted in an increase in DNA-degrading activity with an apparent molecular mass of 15 kDa on a substrate gel. This DNA-degrading activity was entirely calcium dependent and was blocked by the endonuclease inhibitor aurintricarboxylic acid. The protein extract from tubules subjected to hypoxia/reoxygenation cleaved intact nuclear DNA obtained from normal proximal tubules into small fragments, which further supports the presence of endonuclease activity. Despite unequivocal evidence of endonuclease activation, the morphologic features of apoptosis, including chromatin condensation, were not observed by light and electron microscopy. Endonuclease inhibitors, aurintricarboxylic acid and Evans blue, provided complete protection against DNA damage induced by hypoxia/reoxygenation but only partial protection against cell death. Taken together, our data provide strong evidence for a role of endonuclease activation as an early event, which is entirely responsible for the DNA damage and partially responsible for the cell death that occurs during hypoxia/reoxygenation injury. Our data also indicate that in hypoxia/reoxygenation injury endonuclease activation and DNA fragmentation occur without the morphological features of apoptosis.

[Admittatur of Ebenezer Storer (Harvard AB 1747)], 1743

Marbled paper-covered notebook with a handwritten copy of the 1734 College laws in English prepared by Harvard undergraduate Ebenezer Storer and signed by President Edward Holyoke, Fellow Henry Flynt, Tutor Belcher Hancock, Fellow Joseph Mayhew, and Tutor Thomas Marsh on September 11, 1743.

Catalogue of freshmen, 1747

List of the matriculating members of the Harvard Class of 1751.

The mammalian fauna of Warwasi rock shelter, West-central Iran / Priscilla F. Turnbull --

v.33:no.8(1975)

The Summary of a Course of Experimental Philosophical Lectures, 1746-1747

Leather hardcover notebook containing a handwritten copy of John Winthrop's course of experimental and philosophical lectures presented between March 10, 1746 and June 16, 1746. The first one-hundred pages of the volume are divided into twenty chapters which were presented in thirty-three lectures. The chapters contain text and diagrams on mechanical powers, the lever, the pulley, the axis in peritrochio, the inclined plane, the wedge, the screw, compound engines, the laws of motion, gravity, attraction of cohesion, the power of repulsion, magnetism, fluids, electricity, opticks, and astronomy. There is a five-page addenda to the course summary added in 1747, and a sixty-page text titled "The Method of Astronomical calculations" containing thirteen problems related to calculating distances with a list of astronomical characters, and followed with charts related to the eclipse of Jupiter's satellites.

Diary of Henry Flynt, 1723-1747

This diary, effectively a commonplace book, documents Flynt's daily activities and personal reflections from 1723 to 1747. Many entries concern his dealings with family members, business associates, acquaintances, ministers, and political officials. The diary includes a list of books Flynt loaned to others from 1723 to 1743 and detailed financial entries from 1724 to 1747. These entries provide information about the costs of goods and services, as well as Flynt's consumption habits; they detail where he traveled, what he ate and drank (including, apparently, many pounds of almonds), what he read, and many other aspects of daily life. The diary also contains entries related to Flynt's land holdings and other investments, as well as copies of meeting minutes from several sessions of the Harvard Board of Overseers.

Certification of payment from Oliver Prescott, 1747 September 23

Handwritten certificate of payment for a new red and plaid gown purchased by Harvard sophomore Oliver Prescott in 1747. The certificate is witnessed by upperclassmen Artemas Ward, Jacob Cushing, and Timothy Pond.

Deed of sale, 1747 October 21

Deed between grantor Joshua Stratton of Watertown and grantee Andrew Bordman for Cambridge property bordered by lands owned by Isaac Holdin and Andrew Bordman.

Bergen op Zoom, Netherlands, ca. 1747 (Image 1 of 2) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Plan de la forte ville de Berguen op den Zoom avec ses lignes, attaques, et le Fort Steenberguen, desinè par Albert d'Herbort, Ingenieur. It was published by publié par Matthieu Seutter, Geogr. Imperial ca. 1747. Scale [ca. 1:20,000]. Covers Bergen op Zoom, Netherlands. This layer is image 1 of 2 total images of the two sheet source map, representing the southern portion of the map. Map in French. The image inside the map neatline is georeferenced to the surface of the earth and fit to the RD_New (Rijksdriehoekstelsel), GCS Amersfoort coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as drainage, roads, villages and other human settlements, fortifications, lines of fire, ground cover, and more. Relief shown pictorially and by hachures. Includes index.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.