From concept to commercialization; a study of the R & D budget allocation process.

"Conducted by three Sloan fellows, members of the 1960-61 Stanford-Sloan program."

Recollections of the revolution and the empire : from the French of the "Journal d'une femme de cinquante ans," /

"Biographical notes": p. 417-419.

FRAP analysis of nucleocytoplasmic dynamics of the vitamin D receptor splice variant VDRB1: preferential targeting to nuclear speckles

Although the key components of the cellular nuclear transport machinery have largely been characterized through extensive efforts in recent years, in vivo measurements of the kinetics of nuclear protein import/export are patently few. The present study applies the approach of FRAP (fluorescence recovery after photobleaching) to examine the nucleocytoplasmic flux of a novel human VDRB1 (vitamin D receptor B I) isoform in living cells. Through an N-terminal extension containing a consensus nuclear targeting sequence, VDRB1 is capable of localizing in nuclear speckles adjacent to SC-35 (35 kDa splicing component)containing speckles as well as in the nucleoplasm, dependent on ligand. Investigation of VDRB1 nucleocytoplasmic transport using FRAP indicates for the first time that the VDRB1 has a serum-modulated, active nuclear-import mechanism. There is no evidence of an efficient, active export mechanism for VDRB1, probably as a result of nuclear retention. VDRB1 nuclear import in the absence of serum occurred more rapidly and to a greater extent to nuclear speckles compared with import to other nuclear sites. This preferential transport from the cytoplasm to and accumulation within nuclear speckles is consistent with the idea that the latter represent dynamic centres of VDRB1 interaction with other nuclear proteins. The results are consistent with the existence of specialized pathways to target proteins to nuclear subdomains.

Cross-cultural confirmatory factor analysis of the CES-D in Spanish and Mexican dementia caregivers

The Center for Epidemiologic Studies-Depression Scale (CES-D) is the most frequently used scale for measuring depressive symptomatology in caregiving research. The aim of this study is to test its construct structure and measurement equivalence between caregivers from two Spanish-speaking countries. Face-to-face interviews were carried out with 595 female dementia caregivers from Madrid, Spain, and from Coahuila, Mexico. The structure of the CES-D was analyzed using exploratory and confirmatory factor analysis (EFA and CFA, respectively). Measurement invariance across samples was analyzed comparing a baseline model with a more restrictive model. Significant differences between means were found for 7 items. The results of the EFA clearly supported a four-factor solution. The CFA for the whole sample with the four factors revealed high and statistically significant loading coefficients for all items (except item number 4). When equality constraints were imposed to test for the invariance between countries, the change in chi-square was significant, indicating that complete invariance could not be assumed. Significant between-countries differences were found for three of the four latent factor mean scores. Although the results provide general support for the original four-factor structure, caution should be exercised on reporting comparisons of depression scores between Spanish-speaking countries.

Homologues of the maize rust resistance gene Rp1-D are genetically associated with a major rust resistance QTL in sorghum

As part of a comparative mapping study between sugarcane and sorghum, a sugarcane cDNA clone with homology to the maize Rp1-D rust resistance gene was mapped in sorghum. The cDNA probe hybridised to multiple loci, including one on sorghum linkage group (LG) E in a region where a major rust resistance QTL had been previously mapped. Partial sorghum Rp1-D homologues were isolated from genomic DNA of rust-resistant and -susceptible progeny selected from a sorghum mapping population. Sequencing of the Rp1-D homologues revealed five discrete sequence classes: three from resistant progeny and two from susceptible progeny. PCR primers specific to each sequence class were used to amplify products from the progeny and confirmed that the five sequence classes mapped to the same locus on LG E. Cluster analysis of these sorghum sequences and available sugarcane, maize and sorghum Rp1-D homologue sequences showed that the maize Rp1-D sequence and the partial sugarcane Rp1-D homologue were clustered with one of the sorghum resistant progeny sequence classes, while previously published sorghum Rp1-D homologue sequences clustered with the susceptible progeny sequence classes. Full-length sequence information was obtained for one member of a resistant progeny sequence class ( Rp1-SO) and compared with the maize Rp1-D sequence and a previously identified sorghum Rp1 homologue ( Rph1-2). There was considerable similarity between the two sorghum sequences and less similarity between the sorghum and maize sequences. These results suggest a conservation of function and gene sequence homology at the Rp1 loci of maize and sorghum and provide a basis for convenient PCR-based screening tools for putative rust resistance alleles in sorghum.

Identification of the fungal catabolic D-galacturonate pathway

Pectin is a natural polymer consisting mainly of D-galacturonic acid monomers. Microorganisms living on decaying plant material can use D-galacturonic acid for growth. Although bacterial pathways for D-galacturonate catabolism had been described previously, no eukaryotic pathway for D-galacturonate catabolism was known at the beginning of this work. The aim of this work was to identify such a pathway. In this thesis the pathway for D-galacturonate catabolism was identified in the filamentous fungus Trichoderma reesei. The pathway consisted of four enzymes: NADPH-dependent D-galacturonate reductase (GAR1), L-galactonate dehydratase (LGD1), L-threo-3-deoxy-hexulosonate aldolase (LGA1) and NADPH-dependent glyceraldehyde reductase (GLD1). In this pathway D-galacturonate was converted to pyruvate and glycerol via L-galactonate, L-threo-3-deoxy-hexulosonate and L-glyceraldehyde. The enzyme activities of GAR1, LGD1 and LGA1 were present in crude mycelial extract only when T. reesei was grown on D-galacturonate. The activity of GLD1 was equally present on all the tested carbon sources. The corresponding genes were identified either by purifying and sequencing the enzyme or by expressing genes with homology to other similar enzymes in a heterologous host and testing the activities. The new genes that were identified were expressed in Saccharomyces cerevisiae and resulted in active enzymes. The GAR1, LGA1 and GLD1 were also produced in S. cerevisiae as active enzymes with a polyhistidine-tag, and purified and characterised. GAR1 and LGA1 catalysed reversible reactions, whereas only the forward reactions were observed for LGD1 and GLD1. When gar1, lgd1 or lga1 was deleted in T. reesei the deletion strain was unable to grow with D-galacturonate as the only carbon source, demonstrating that all the corresponding enzymes were essential for D-galacturonate catabolism and that no alternative D-galacturonate pathway exists in T. reesei. A challenge for biotechnology is to convert cheap raw materials to useful and more valuable products. Filamentous fungi are especially useful for the conversion of pectin, since they are efficient producers of pectinases. Identification of the fungal D-galacturonate pathway is of fundamental importance for the utilisation of pectin and its conversion to useful products.

Structure, genomic DNA typing, and kinetic characterization of the D allozyme of placental alkaline phosphatase (PLAP/ALPP)

The D allozyme of placental alkaline phosphatase (PLAP) displays enzymatic properties at variance with those of the common PLAP allozymes. We have deduced the amino acid sequence of the PLAP D allele by PCR cloning of its gene, ALPP Two coding substitutions were found in comparison With the cDNA of the common PLAP F allele, i.e., 692C>G and 1352A>G, which translate into a P209R and E429G substitution. A single nucleotide primer extension (SNuPE) assay was developed using PCR primers that enable the amplification of a 1.9 kb PLAP fragment. Extension primers were then used on this PCR fragment to detect the 692C>G and 1352A>G substitution. The SNuPE assay on these two nucleotide substitutions enabled us to distinguish the PLAP F and D alleles from the PLAP S/I alleles. Functional studies on the D allozyme were made possible by constructing and expressing a PLAP D cDNA, i.e., [Arg209, Gly429] PLAP, into wildtype Chinese hamster ovary cells. We determined the k(cat) and K-m, of the PLAP S, F. and D allozymes using the non,physiological substrate p-nitrophenylphosphate at an optimal pH (9.8) as well as two physiological substrates, i.e., pyridoxal-5'-phosphate and inorganic pyrophosphate at physiological pH (7.5). We found that the biochemical properties of the D allozyme of PLAP are significantly different from those of the common PLAP allozymes. These biochemical findings suggest that a suboptimal enzymatic function by the PLAP D allozyme may be the basis for the apparent negative selective pressure of the PLAP D allele. The development of the SNuPE assay will enable us to test the hypothesis that the PLAP D allele is subjected to intrauterine selection by examining genomic DNA from statistically informative population samples. Hum Mutat 19:258-267, 2002. (C) 2002 Wiley-Liss, Inc.

Surface sediment analyses, bivalve and gastropod taxa of marine sediments from the Banc d'Arguin, Mauritania

High-nutrient tropical carbonate systems are known to produce sediments that, in terms of skeletal composition, are reminiscent of their extra-tropical counterparts. Such carbonate systems and associated carbonate grain assemblages in the tropics are rare in the present-day world. Nonetheless, it is crucial to gain a better understanding of those ecosystems, including their drivers and players because such settings potentially represent models for ancient depositional systems as well as for predicted future environmental conditions. One of the modern occurrences of eutrophic tropical carbonate systems is the northern Mauritanian Shelf. The marine environment is characterized by an eastern boundary upwelling system that pushes cool and nutrient-rich intermediate waters onto a wide epicontinental platform (Golfe d'Arguin) where the waters warm up to tropical temperatures. The resulting facies is mixed carbonate-siliciclastic with a dominant foramol association grading into bimol and barnamol grain assemblages in the shallowest areas forming the Banc d'Arguin. Besides this cool water-related heterozoan association, the carbonate sediment is characterized by tropical molluskan species, while chlorozoan biota (e.g., corals and algal symbiont-bearing foraminifers) are entirely absent. We here present a first comprehensive facies analysis of this model example of eutrophic tropical carbonates. Furthermore, we reconstruct the loci of carbonate production and provide a conclusive depositional model of the Banc d'Arguin that received little attention to date due to its poorly accessible nature.

Pollen analysis of soil samples from the A.D. 79 level. Boscoreale, Oplontis, and Pompeii

Fifty samples of Roman time soil preserved under the thick ash layer of the A.D.79 eruption of Mt Vesuvius were studied by pollen analysis: 33 samples from a former vineyard surrounding a Villa Rustica at Boscoreale (excavation site 40 x 50 m), 13 samples taken along the 60 m long swimming pool in the sculpture garden of the Villa of Poppaea at Oplontis, and four samples from the formal garden (12.4 x 17.5 m) of the House of the Gold Bracelet in Pompeii. To avoid contamination with modern pollen all samples were taken immediately after uncovering a new portion of the A.D. 79 soil. For comparison also samples of modern Italian soils were studied. Using standard methods for pollen preparation the pollen content of 15 of the archaeological samples proved to be too little to reach a pollen sum of more than 100 grains. The pollen spectra of these samples are not shown in the pollen tables. (Flotation with a sodium tungstate solution, Na2WO4, D = 2.05, following treatment with HCl and NaOH would probably have given a somewhat better result. This method was, however, not available as too expensive at that time.) Although the archaeological samples were taken a few meters apart their pollen values differ very much from one sample to the other. E.g., at Boscoreale (SW quarter). the pollen values of Pinus range from 1.5 to 54.5% resp. from 1 to 244 pine pollen grains per 1 gram of soil, the extremes even found under pine trees. Vitis pollen was present in 7 of the 11 vineyard samples from Boscoreale (NE quarter) only. Although a maximum of 21.7% is reached, the values of Vitis are mostly below 1.5%. Even the values of common weeds differ very much, not only at Boscoreale, but also at the other two sites. The pollen concentration values show similar variations: 3 to 3053 grains and spores were found in 1 g of soil. The mean value (290) is much less than the number of pollen grains, which would fall on 1 cm2 of soil surface during one year. In contrast, the pollen and spore concentrations of the recent soil samples, treated in exactly the same manner, range from 9313 to almost 80000 grains per 1 g of soil. Evidently most of the Roman time pollen has disappeared since its deposition, the reasons not being clear. Not even species which are known to have been cultivated in the garden of Oplontis, like Citrus and Nerium, plant species with easily distinguishable pollen grains, could be traced by pollen analysis. The loss of most of the pollen grains originally contained in the soil prohibits any detailed interpretation of the Pompeian pollen data. The pollen counts merely name plant species which grew in the region, but not necessarily on the excavated plots.

A Case Study of Geometric Modelling via 3-D Point Interpolation for the Bathymetry of the Rabasa Lakes (Alicante, Spain)

The interpolation of points by means of Information Technology programs appears as a technical tool of some relevancy in the hydrogeology in general and in the study of the humid zones in particular. Our approach has been the determination of the 3-D geometry of the humid zones of major depth of the Rabasa Lakes. To estimate the topography of the lake bed, we proceed to acquire information in the field by means of sonar and GPS equipment. A total of 335 points were measured both on the perimeter and in the lake bed. In a second stage, this information was used in a kriging program to obtain the bathymetry of the wetland. This methodology is demonstrated as one of the most reliable and cost-efficient for the 3-D analysis of this type of water masses. The bathymetric study of the zone allows us to characterize the mid- and long-term hydrological evolution of the lakes by means of depth-area-volume curves.

(Tab. XIII.1) TOC content, carbonate content, carbonate mineralogy, percentage of total carbonate, aragonite, low magnesium calcium, and high magnesium calcium, and grain-size contents from samples from the Golfe d'Arguin and from the Western Sahara

Modern carbonate sedimentation takes place on the northern Mauritanian shelf (20°N), where typical tropical components (e.g. hermatypic reefs, calcareous green algae) are absent. Such deposits are reminiscent of extratropical sediment in the geological record. The tropical open shelf of Mauritania is influenced by large siliciclastic dust input and upwelling, highly fertilizing the ocean, as well as strongly limiting the light penetration. In this context, temperature does not appear to be the steering factor of carbonate production. This thesis describes the depositional system of the Golfe d'Arguin off Mauritania and focuses on environmental conditions that control the depositional pattern, in particular carbonate production. The description of this modern analogue provides a tool for paleoenvironmental interpretation of ancient counterparts. The Golfe d'Arguin is a broad shallow shelf comprising extensive shoals (<10 m water depth; i.e. the Banc d'Arguin) on the inner shelf where waters warm up. The sediments collected in water depths between 4 and 600 m are characterized by mixed carbonate and siliciclastic (dust) deposits. They vary from clean coarse-grained, almost pure carbonate loose sediments to siliciclastic-dominated fine-grained sediments. The carbonate content and sediment grain size show a north-south decreasing pattern through the Golfe d'Arguin and are controlled by the hydraulic regime influenced by wind-driven surface currents, swell, and tidal currents. The carbonate grain association is heterozoan. Components include abundant molluscs, foraminifers, and worm tubes, as well as barnacles and echinoderms, elements that are also abundant in extratropical sediments. The spatial distribution of the sedimentary facies of the Golfe d'Arguin does not display a depth zonation but rather a mosaic (i.e. patchy distribution). The depth and climatic signatures of the different sedimentary facies are determined by taxonomic and ecological investigations of the carbonate-secreting biota (molluscs and foraminifers). While certain planktonic foraminifers and molluscs represent upwelling elements, other components (e.g. mollusc and benthic foraminifer taxa) demonstrate the tropical origin of the sediment. The nutrient-rich (and thus also low light-penetration) conditions are reflected in the fact that symbiotic and photosynthetic carbonate-producing organisms (e.g. hermatypic corals) are absent. The Mauritanian deposits represent an environment that is rare in the modern world but might have been more common in the geological past when global temperatures were higher. Taxonomic and ecological studies allow for distinguishing carbonate sediments formed under either tropical high-nutrient or extratropical conditions, thus improving paleoclimate reconstruction.

(Table 1) Quantitative analysis of mollusk taxa from surface sediment samples in the Golfe d'Arguin

Ecological and taxonomic study of the mollusk-rich fauna of the Golfe d'Arguin, North Mauritania, investigates the various environmental influences affecting this tropical shelf. The upwelling of nutrient-rich waters leads to a highly productive environment under tropical conditions. The resulting mixed carbonate-siliciclastic sediment contains a large portion of calcareous components produced by heterotrophic organisms-e.g., mollusks, foraminifers, worms, barnacles-that are reworked on the open shelf. On the basis of mollusk assemblages, six taphocoenoses are defined, all being characterized by a mixed fauna of tropical (e.g., Tellina densestriata), subtropical (e.g., Macoma cumana) and temperate (e.g., Spisula subtruncata) species. Differences between the assemblages are related to the medium-grain size ranging from mud to gravel-that results from local hydrodynamic conditions and water depth. Among carbonate grains, Donax burnupi shells are very abundant in the swell-exposed, northern part of the Golfe d'Arguin and reflect the tropical to subtropical, high-energy, and high-nutrient waters. Mollusk assemblages are demonstrated to be a sensitive tool for deciphering complex environmental conditions in sedimentary archives.

Homologues of the maize rust resistance gene Rp1-D are genetically associated with a major rust resistance QTL in sorghum

As part of a comparative mapping study between sugarcane and sorghum, a sugarcane cDNA clone with homology to the maize Rp1-D rust resistance gene was mapped in sorghum. The cDNA probe hybridised to multiple loci, including one on sorghum linkage group (LG) E in a region where a major rust resistance QTL had been previously mapped. Partial sorghum Rp1-D homologues were isolated from genomic DNA of rust-resistant and -susceptible progeny selected from a sorghum mapping population. Sequencing of the Rp1-D homologues revealed five discrete sequence classes: three from resistant progeny and two from susceptible progeny. PCR primers specific to each sequence class were used to amplify products from the progeny and confirmed that the five sequence classes mapped to the same locus on LG E. Cluster analysis of these sorghum sequences and available sugarcane, maize and sorghum Rp1-D homologue sequences showed that the maize Rp1-D sequence and the partial sugarcane Rp1-D homologue were clustered with one of the sorghum resistant progeny sequence classes, while previously published sorghum Rp1-D homologue sequences clustered with the susceptible progeny sequence classes. Full-length sequence information was obtained for one member of a resistant progeny sequence class (Rp1-SO) and compared with the maize Rp1-D sequence and a previously identified sorghum Rp1 homologue (Rph1-2). There was considerable similarity between the two sorghum sequences and less similarity between the sorghum and maize sequences. These results suggest a conservation of function and gene sequence homology at the Rp1 loci of maize and sorghum and provide a basis for convenient PCR-based screening tools for putative rust resistance alleles in sorghum.