Protective immune responses induced by secretion of a chimeric soluble protein from a recombinant Mycobacterium bovis bacillus Calmette-Guérin vector candidate vaccine for human immunodeficiency virus type 1 in small animals.

A recombinant Mycobacterium bovis bacillus Calmette-Guérin (BCG) vector-based vaccine that secretes the V3 principal neutralizing epitope of human immunodeficiency virus (HIV) could induce immune response to the epitope and prevent the viral infection. By using the Japanese consensus sequence of HIV-1, we successfully constructed chimeric protein secretion vectors by selecting an appropriate insertion site of a carrier protein and established the principal neutralizing determinant (PND)-peptide secretion system in BCG. The recombinant BCG (rBCG)-inoculated guinea pigs were initially screened by delayed-type hypersensitivity (DTH) skin reactions to the PND peptide, followed by passive transfer of the DTH by the systemic route. Further, immunization of mice with the rBCG resulted in induction of cytotoxic T lymphocytes. The guinea pig immune antisera showed elevated titers to the PND peptide and neutralized HIVMN, and administration of serum IgG from the vaccinated guinea pigs was effective in completely blocking the HIV infection in thymus/liver transplanted severe combined immunodeficiency (SCID)/hu or SCID/PBL mice. In addition, the immune serum IgG was shown to neutralize primary field isolates of HIV that match the neutralizing sequence motif by a peripheral blood mononuclear cell-based virus neutralization assay. The data support the idea that the antigen-secreting rBCG system can be used as a tool for development of HIV vaccines.

Transcription factor GATA-1 permits survival and maturation of erythroid precursors by preventing apoptosis.

The transcription factor GATA-1 recognizes a consensus motif present in regulatory regions of numerous erythroid-expressed genes. Mouse embryonic stem cells lacking GATA-1 cannot form mature red blood cells in vivo. In vitro differentiation of GATA-1- embryonic stem cells gives rise to a population of committed erythroid precursors that exhibit developmental arrest and death. We show here that the demise of GATA-1- erythroid cells is accompanied by several features characteristics of apoptosis. This process occurs despite normal expression of all known GATA target genes examined, including the erythropoietin receptor, and independent of detectable accumulation of the tumor suppressor protein p53. Thus, in addition to its established role in regulating genes that define the erythroid phenotype, GATA-1 also supports the viability of red cell precursors by suppressing apoptosis. These results illustrate the multifunctional nature of GATA-1 and suggest a mechanism by which other hematopoietic transcription factors may ensure the development of specific lineages.

A structural motif for the voltage-gated potassium channel pore.

Mutation studies have identified a region of the S5-S6 loop of voltage-gated K+ channels (P region) responsible for teraethylammonium (TEA) block and permeation/selectivity properties. We previously modeled a similar region of the Na+ channel as four beta-hairpins with the C strands from each of the domains forming the external vestibule and with charged residues at the beta-turns forming the selectivity filter. However, the K+ channel P region amino acid composition is much more hydrophobic in this area. Here we propose a structural motif for the K+ channel pore based on the following postulates (Kv2.1 numbering). (i) The external TEA binding site is formed by four Tyr-380 residues; P loop residues participating in the internal TEA binding site are four Met-371 and Thr-372 residues. (ii) P regions form extended hairpins with beta-turns in sequence ITMT. (iii) only C ends of hairpins form the inner walls of the pore. (iv) They are extended nonregular strands with backbone carbonyl oxygens of segment VGYGD facing the pore with the conformation BRLRL. (v) Juxtaposition of P loops of the four subunits forms the pore. Fitting the external and internal TEA sites to TEA molecules predicts an hourglass-like pore with the narrowest point (GYG) as wide as 5.5 A, suggesting that selectivity may be achieved by interactions of carbonyls with partially hydrated K+. Other potential cation binding sites also exist in the pore.

A chloroplast processing enzyme involved in precursor maturation shares a zinc-binding motif with a recently recognized family of metalloendopeptidases.

Nuclear-encoded proteins targeted to the chloroplast are typically synthesized with N-terminal transit peptides which are proteolytically removed upon import. Structurally related proteins of 145 and 143 kDa copurify with a soluble chloroplast processing enzyme (CPE) that cleaves the precursor for the major light-harvesting chlorophyll a/b binding protein and have been implicated in the maturation of the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase and acyl carrier protein. The 145- and 143-kDa proteins have not been found as a heterodimer and thus may represent functionally independent isoforms encoded by separate genes. Here we describe the primary structure of a 140-kDa polypeptide encoded by cDNAs isolated by using antibodies raised against the 145/143-kDa doublet. The 140-kDa polypeptide contains a transit peptide, and strikingly, a His-Xaa-Xaa-Glu-His zinc-binding motif that is conserved in a recently recognized family of metalloendopeptidases, which includes Escherichia coli protease III, insulin-degrading enzyme, and subunit beta of the mitochondrial processing peptidase. Identity of 25-30%, concentrated near the N terminus of the 140-kDa polypeptide, is found with these proteases. Expression of CPE in leaves is not light dependent. Indeed, transcripts are present in dark-grown plants, and the 145/143-kDa doublet and proteolytic activity are both found in etioplasts, as well as in root plastids. Thus, CPE appears to be a necessary component of the import machinery in photosynthetic and nonphotosynthetic tissues, and it may function as a general stromal processing peptidase in plastids.

The alpha 5 beta 1 integrin supports survival of cells on fibronectin and up-regulates Bcl-2 expression.

Anchorage-dependent cells that are prevented from attaching to an extracellular matrix substrate stop proliferating and may undergo apoptosis. Cell adhesion to a substrate is mediated by the integrin family of cell surface receptors, which are known to elicit intracellular signals upon cell adhesion. We show here that Chinese hamster ovary cells expressing the alpha 5 beta 1 integrin, which is a fibronectin receptor, do not undergo apoptosis upon serum withdrawal when the cells are plated on fibronectin. However, the alpha v beta 1 integrin, which is also a fibronectin receptor and binds fibronectin on the same RGD motif as alpha 5 beta 1, did not prevent apoptosis on fibronectin of the same cells. The cytoplasmic domain of the integrin alpha 5 subunit was required for the alpha 5 beta 1-mediated cell survival on fibronectin. The fibronectin-mediated survival effect appeared to be independent of the level of tyrosine phosphorylation of the focal adhesion kinase, which is induced by integrin-mediated cell attachment. The expression of the Bcl-2 protein, which counteracts apoptosis, was elevated in cells attaching to fibronectin through alpha 5 beta 1; cells attaching through alpha v beta 1 survived only if exogenous Bcl-2 was provided. Thus, alpha 5 beta 1, but not the closely related alpha v beta 1 integrin, appears to suppress apoptotic cell death through the Bcl-2 pathway.

Two nuclear localization signals present in the basic-helix 1 domains of MyoD promote its active nuclear translocation and can function independently.

MyoD, a member of the family of helix-loop-helix myogenic factors that plays a crucial role in skeletal muscle differentiation, is a nuclear phosphoprotein. Using microinjection of purified MyoD protein into rat fibroblasts, we show that the nuclear import of MyoD is a rapid and active process, being ATP and temperature dependent. Two nuclear localization signals (NLSs), one present in the basic region and the other in the helix 1 domain of MyoD protein, are demonstrated to be functional in promoting the active nuclear transport of MyoD. Synthetic peptides spanning these two NLSs and biochemically coupled to IgGs can promote the nuclear import of microinjected IgG conjugates in muscle and nonmuscle cells. Deletion analysis reveals that each sequence can function independently within the MyoD protein since concomittant deletion of both sequences is required to alter the nuclear import of this myogenic factor. In addition, the complete cytoplasmic retention of a beta-galactosidase-MyoD fusion mutant protein, double deleted at these two NLSs, argues against the existence of another functional NLS motif in MyoD.

(Figure 1) Silicoflagellates at DSDP Hole 69-504

Diverse and abundant late Miocene to Pleistocene silicoflagellates at DSDP Site 504 can be correlated by tropical biostratigraphic zones and relative paleotemperature values to eastern tropical Pacific reference site DSDP 503A farther to the west. Early Pliocene assemblages, which were poorly known until now, are present and can be correlated locally between DSDP Holes 504, 503A, and 495, using species events associated with the new Dictyocha pulchella Subzone and Dictyocha angulata Subzone. Silicoflagellate relative paleotemperature values show major warming at 4.7 to 5.0 Ma (Cores 45-48), 3.4 to 3.8 Ma (Cores 32-33), 1.5 to 1.7 Ma (Cores 12-16), and 0.5 to 0.8 Ma (Cores 3-6). Major coolings occurred at 5.0 to 5.1 Ma (Core 51), 3.9 to 4.4 Ma (Cores 38-44), and 1.0 to 1.3 Ma (Cores 8-10). The appearance of Dictyocha longa is proposed to replace the asperoid/fibuloid ratio reversal as the bottom of the Dictyocha fibula Zone, because the non-evolutionary ratio reverses several times in the upper Miocene of Hole 503A, and at least once in Hole 504. Three new Pliocene silicoflagellates are defined: Dictyocha concinna Bukry, n. sp., D. helix Bukry, n. sp., and D. tamarae Bukry, n. sp.

(Table 1) Paleomagnetic properties of specimens from DSDP Holes 69-504B and 70-504B

Paleomagnetic and rock magnetic measurements of basalt specimens from DSDP Hole 504B, associated with the Costa Rica Rift, have a mean natural remanence intensity (Jn) between 5 and 10 x 10**-3 gauss, consistent with the presence of a magnetized layer that is 0.5 to 1 km thick, which produces the observed magnetic anomalies. A mean Koenigsberger ratio (Qn) greater than 10 indicates that the remanence dominates the magnetic signal of the drilled section. The susceptibility (x) increases with depth, and the median demagnetizing field (MDF) decreases with increasing depth in Hole 504B, congruent with the downhole increase in the relative abundance of massive flow units. Hole 504B is composed of at least 12 units with distinct stable average inclinations (Is), which probably represent extrusion at times of different geomagnetic field directions and possibly also the effects of faulting. The thickness of basalt associated with these inclination units varies from less than 9 meters to possibly as much as 160 meters. Two relatively thick magnetic units (40 m and 45 m, separated by 100 m) have anomalously high Is values of -53° and -63°, in contrast with the near zero inclinations expected for the equatorial latitude of Site 504. For this reason and because the average inclination of all the magnetic units is skewed to a negative value, it might be that the entire section at Hole 504B was tilted by approximately 30°.

(Table 1) K-Ar age and inert-gas-isotopes at DSDP Holes 68-501, 69-504B and 69-505B

Conventional K-Ar ages have been determined and inert-gas abundances have been measured on representative samples of altered rocks from Deep Sea Drilling Project Holes 501, 504B, and 505B in an attempt to correlate their degree of alteration with inert-gas and K-Ar data. Samples taken from the first 60 meters below the sediment/basalt interface give significantly higher ages than would be expected from the magnetic stratigraphy, though at greater depths the calculated ages are in broad agreement with the expected age. The inert gas ratios 20Ne/36Ar, 36Ar/84Kr, and 84Kr/130Xe also show a marked discontinuity at the 60-meter depth, and all these effects are interpreted as being a consequence of low-temperature alteration produced by burial metamorphism and by interaction with sea water (halmyrolysis).

(Table 1) Element concentrations in detrital sediments of ODP Site 184-1148

The chemical index of alteration (CIA) and elemental ratios that are sensitive to chemical weathering, such as Ca/Ti, Na/Ti, Al/Ti, Al/Na, Al/K, and La/Sm, were analyzed for detrital sediments at Ocean Drilling Program Site 1148 from the northern South China Sea to reveal information of chemical weathering in the source regions during the early Miocene. High CIA values of ~80, coupled with high Al/Ti and Al/Na and low Na/Ti and Ca/Ti, are observed for the sediments at ~23 Ma, indicating a high chemical weathering intensity in the north source region, i.e., south China. This was followed by gradual decreases in Al/Ti, Al/Na, La/Sm, and Al/K ratios, as well as the CIA values, and increases in Ca/Ti and Na/Ti ratios. These records together with other paleoclimate proxies, such as black carbon d13C and benthic foraminifer d18O, give reliable information on the climate changes in south China. Our results show that the climate in south China was warm and humid in the early Miocene (~23 Ma) according to the chemical weathering records. The humidity in south China decreased from the early Miocene to Present with several fluctuations centering at approximately 15.7 Ma, 8.4 Ma, and 2.5 Ma, coincident with the global cooling since the middle Miocene. These climate changes implied that the summer east Asian monsoon has dramatically affected south China in the early Miocene, whereas the influence of the summer monsoon on this region has decreased continuously since that time, probably because of the intensification of the winter monsoon. Such an evolution for the east Asian monsoon is different from that for the Indian monsoon.

Bowl with Kufic inscription and tree motif

Glass, Islamic, Fatamid; 4 7/32 in. x 6 1/32 in.; blown glass and stained glass with copper and silver stains

Édouard Pailleron. Théātre complet. 1-4.

1. Le Parasite. Le Mur mitoyen. La dernier Quartier. Le second Mouvement. Le Monde où l'on s'amuse. [1909.] 2. Les faux Ménages. L'autre Motif. Hélene. Petite Pluie. [1910.] L'Āge ingrat. Le Chevalier Trumeau. L'Etincelle. Pendant lebal. Le Monde où l'on s'ennuie. [1911.] 4. Le Narcotique. La Souris. Cabotins. Un grand Enterrement. [1912.]

Utvaldar smá-søgur, almenníngai til-fródleiks og skémtumar.

Imperfect: pt. 1, cover-title, p. i-iv wanting.

Operator's and organizational maintenance manual including repair parts and special tools lists : simulator, radio frequency SM-442A/GRC.

"August 1967."