Sequence analysis of the cis-regulatory regions of the bithorax complex of Drosophila.

The bithorax complex (BX-C) of Drosophila, one of two complexes that act as master regulators of the body plan of the fly, has now been entirely sequenced and comprises approximately 315,000 bp, only 1.4% of which codes for protein. Analysis of this sequence reveals significantly overrepresented DNA motifs of unknown, as well as known, functions in the non-protein-coding portion of the sequence. The following types of motifs in that portion are analyzed: (i) concatamers of mono-, di-, and trinucleotides; (ii) tightly clustered hexanucleotides (spaced < or = 5 bases apart); (iii) direct and reverse repeats longer than 20 bp; and (iv) a number of motifs known from biochemical studies to play a role in the regulation of the BX-C. The hexanucleotide AGATAC is remarkably overrepresented and is surmised to play a role in chromosome pairing. The positions of sites of highly overrepresented motifs are plotted for those that occur at more than five sites in the sequence, when < 0.5 case is expected. Expected values are based on a third-order Markov chain, which is the optimal order for representing the BXCALL sequence.

SPC3, a synthetic peptide derived from the V3 domain of human immunodeficiency virus type 1 (HIV-1) gp120, inhibits HIV-1 entry into CD4+ and CD4- cells by two distinct mechanisms.

The third variable region (V3 loop) of gp120, the HIV-1 surface envelope glycoprotein, plays a key role in HIV-1 infection and pathogenesis. Recently, we reported that a synthetic multibranched peptide (SPC3) containing eight V3-loop consensus motifs (GPGRAF) inhibited HIV-1 infection in both CD4+ and CD4- susceptible cells. In the present study, we investigated the mechanisms of action of SPC3 in these cell types--i.e., CD4+ lymphocytes and CD4- epithelial cells expressing galactosylceramide (GalCer), an alternative receptor for HIV-1 gp120. We found that SPC3 was a potent inhibitor of HIV-1 infection in CD4+ lymphocytes when added 1 h after initial exposure of the cells to HIV-1, whereas it had no inhibitory effect when present only before and/or during the incubation with HIV-1. These data suggested that SPC3 did not inhibit the binding of HIV-1 to CD4+ lymphocytes but interfered with a post-binding step necessary for virus entry. In agreement with this hypothesis, SPC3 treatment after HIV-1 exposure dramatically reduced the number of infected cells without altering gp120-CD4 interaction or viral gene expression. In contrast, SPC3 blocked HIV-1 entry into CD4-/GalCer+ human colon epithelial cells when present in competition with HIV-1 but had no effect when added after infection. Accordingly, SPC3 was found to inhibit the binding of gp120 to the GalCer receptor. Thus, the data suggest that SPC3 affects HIV-1 infection by two distinct mechanisms: (i) prevention of GalCer-mediated HIV-1 attachment to the surface of CD4-/GalCer+ cells and (ii) post-binding inhibition of HIV-1 entry into CD4+ lymphocytes.

Primary structure of hepatocyte nuclear factor/forkhead homologue 4 and characterization of gene expression in the developing respiratory and reproductive epithelium.

Members of the winged helix/forkhead family of transcription factors are believed to play a role in cell-specific gene expression. A cDNA encoding a member of this family of proteins, termed hepatocyte nuclear factor/forkhead homologue 4 (HFH-4), has been isolated from rat lung and rat testis cDNA libraries. This cDNA contains an open reading frame of 421 amino acids with a conserved DNA binding domain and several potential transactivating regions. During murine lung development, a single species of HFH-4-specific transcript (2.4 kb long) is first detected precisely at the start of the late pseudoglandular stage (embryonic day 14.5) and, by in situ hybridization, is specifically localized to the proximal pulmonary epithelium. The unique temporal and spatial pattern of HFH-4 gene expression in the developing lung defines this protein as a marker for the initiation of bronchial epithelial cell differentiation and suggests that it may play an important role in cell fate determination during lung development. In addition to expression in the pulmonary epithelium, RNA blot analysis reveals 2.4-kb HFH-4 transcripts in the testis and oviduct. By using mice with genetic defects in spermatogenesis, HFH-4 expression in the testis is found to be associated with the appearance of haploid germ cells and in situ hybridization studies indicate that HFH-4 expression is confined to stages I-VII of spermatogenesis. This pattern of HFH-4 gene expression during the early stages of differentiation of haploid germ cells suggests that HFH-4 may play a role in regulating stage-specific gene expression and cell-fate determination during lung development and in spermatogenesis.

Glycosylation of the c-Myc transactivation domain.

O-linked N-acetylglucosamine (O-GlcNAc) is an abundant and dynamic posttranslational modification composed of a single monosaccharide, GlcNAc, glycosidically composed of a single monosaccharide, GlcNAc, glycosidically linked to the side-chain hydroxyl of serine or threonine residues. Although O-GlcNAc occurs on a myriad of nuclear and cytoplasmic proteins, only a few have thus far been identified. These O-GlcNAc-bearing proteins are also modified by phosphorylation and form reversible multimeric complexes. Here we present evidence for O-GlcNAc glycosylation of the oncoprotein c-Myc, a helix-loop-helix/leucine zipper phosphoprotein that heterodimerizes with Max and participates in the regulation of gene transcription in normal and neoplastic cells. O-GlcNAc modification of c-Myc is shown by three different methods: (i) demonstration of lectin binding to in vitro translated protein using a protein-protein interaction mobility-shift assay; (ii) glycosidase or glycosyltransferase treatment of in vitro translated protein analyzed by lectin affinity chromatography; and (iii) direct characterization of the sugar moieties on purified recombinant protein overexpressed in either insect cells or Chinese hamster ovary cells. Analyses of serial deletion mutants of c-Myc further suggest that the O-GlcNAc site(s) are located within or near the N-terminal transcription activation/malignant transformation domain, a region where mutations of c-Myc that are frequently found in Burkitt and AIDS-related lymphomas cluster.

Effects of the environment on the electric conductivity of double-stranded DNA molecules

We present a theoretical analysis of the effects of the environment on charge transport in double-stranded synthetic poly(G)-poly(C) DNA molecules attached to two ideal leads. Coupling of the DNA to the environment results in two effects: (i) localization of carrier functions due to static disorder and (ii) phonon-induced scattering of the carriers between the localized states, resulting in hopping conductivity. A nonlinear Pauli master equation for populations of localized states is used to describe the hopping transport and calculate the electric current as a function of the applied bias. We demonstrate that, although the electronic gap in the density of states shrinks as the disorder increases, the voltage gap in the I-V characteristics becomes wider. A simple physical explanation of this effect is provided.

Comment on Rule 14a-8(i)(9), Securities and Exchange Commission, June 30, 2015

The Securities and Exchange Commission has been analyzing its interpretation under Subsection (i)(9) of Rule 14a-8. This provision allows for the exclusion of shareholder proposals that conflict with those submitted by management. The staff has been examining its interpretation since instructed to do so by the chair of the SEC following a no action appeal in a case involving Whole Foods. A number of letters and memos submitted in connection with the review have analyzed the issue. At least one asserted that any change in interpretation required the Commission to go through the process of notice and comment. This letter asserts that notice and comment is not required, relying extensively on Perez v. Mortgage Bankers Association.

Radiological impact of cement, concrete and admixtures in Spain

It has been analyzed samples of portland cement (PC) with and without admixtures, samples of calcium aluminate cement (CAC) with different content of Al2O3 and specimens of concrete made with PC and CAC using High Resolution Gamma Spectrometry. The activity concentration index (I) is much less than 0.5 mSv y-1 for all the concrete specimens according to the Radiation protection document 112 of the European Commission. The PC without admixtures (CEM I 52,5 R) and the PC with addition of limestone (CEM II/BL 32,5 N) also have an I value much lower than 0.5 and the PC with the addition of fly ash and blast furnace slag (CEM IV/B (V) 32,5 N and III/A 42.5 N/SR) have an I value close to 0.6. The I value of the CAC used in the manufacture of structural precast concrete is of the order of 1 mSv y-1. Some of the CAC used in refractory concrete reaches a value close to 2 mSv y-1.

Analysis of halogenated compounds in landfill gas

The authors thank the research project CTQ2008-05520 from the Spanish Ministry of Education and Science and PROMETEO/2009/043/FEDER from the Valencian Community Government (Spain) for the support provided.

A Theme, being the Employment of some Leisure time Nov'r 14 1759 pr Jeremiah Belknap, 1759 November 14

Handwritten composition on two attached notebook pages titled "The Generous man is a blessing to all mankind" composed by Jeremy Belknap and organized as paragraphs labeled: "Proposition," "Reason," Similitude," "Example," "Testimony," and "Conclusion."

Analysis of Luke 14, 1761 October

Two notebook sheets with handwritten summaries in English of Luke 14 beginning, "The contents of this chapter may be reduced to 5 General Division..." The document is a draft with edits and struck-through words, and is dated and signed "Boston, 1[9] October 1761. J. Belknap" with the note, "Read in the Chapel of Harvard College at Even'g Prayers Octob. 1761."

The Death of John the Baptist, [ca. 1761]

Four-page handwritten English composition containing a sketch of the death of John the Baptist beginning "John was imprisoned in the Castle of Machaerus..."

Charter of 1650, 1650 May 31

Original charter dated May 31, 1650. The charter is engrossed on sheepskin and signed by Thomas Dudley.

[Notes from the minutes of the Board of Overseers, ca. 1799]

Small notebook containing handwritten entries of actions for the years 1718-1799 recorded in Volumes I-IV of the Harvard Board of Overseers formal meeting minutes. The entries note the appointment of Committees of inquiry and votes related to salary increases and allowances.

"A list of the books belonging to the late Library of Harvard College that were in the hands of the Overseers, Governors, & Students of the College & escap'd the flames," ca. 1764

This list appears to be the most comprehensive in this series. Although its contents are very similar to those of the list in Folder 2, there are some discrepancies. Entries are arranged by format (folio, quarto, octavo) and include the date the book was "delivered" (loaned), the name of the individual who borrowed it, and its author, title, and volume number. Many of the books had been out of the library for decades prior to the fire, with some loaned out since as early as 1742.

Draft of "A list of the books belonging to the late Library of Harvard College [...]," ca. 1764

This list appears to be a draft of the list in Folder 1. Although most of the entries match, there are some discrepancies.