Transferrinfection. A Highly Efficient Way to Express Gene Constructs in Eukaryotic Cells

No abstract available

High-efficiency receptor-mediated delivery of small and large (48 kilobase gene constructs using the endosome-disruption activity of defective or chemically inactivated adenovirus particles.

One limit to successful receptor-mediated gene delivery is the exit of the endocytosed material from the endosome. We demonstrate here the delivery of marker genes to tissue culture cells using a modification of the receptor-mediated gene delivery technique that exploits the endosomolytic activity of defective adenovirus particles. In particular, greater than 90% of the transfected-cell population is found to express a beta-galactosidase gene, and, most importantly, this high level of expression can be obtained with psoralen-inactivated virus particles. Furthermore, because the delivered gene is not carried within the genome of the adenovirus particle, the size constraints are relieved, and we can, therefore, show the delivery of a 48-kilobase cosmid DNA molecule

Coupling of adenovirus to transferrin-polylysine/DNA complexes greatly enhances receptor-mediated gene delivery and expression of transfected genes.

We are developing efficient methods for gene transfer into tissue culture cells. We have previously shown that coupling of a chimeric adenovirus with polylysine allowed the construction of an adenovirus-polylysine-reporter-gene complex that transferred the transporter gene with great efficiency into HeLa cells. We have now explored simpler, biochemical means for coupling adenovirus to DNA/polylysine complexes and show that such complexes yield virtually 100% transfection in tissue culture cell lines. In these methods adenovirus is coupled to polylysine, either enzymatically through the action of transglutaminase or biochemically by biotinylating adenovirus and streptavidinylating the polylysine moiety. Combination complexes containing DNA, adenovirus-polylysine, and transferrin-polylysine have the capacity to transfer the reporter gene into adenovirus-receptor- and/or transferrin-receptor-rich cells

Propagation en contexte arrière-arc : premiers résultats de la campagne ProFeTi (Bassin Nord-Fidjien, Pacifique SW)

Au centre du bassin Nord-Fidjien, le segment d'accrétion NS, qui se propage vers le Nord aux dépens du segment N15 depuis au moins 1 Ma, a été échantillonné pendant la campagne ProFeTi du NO Alis. Malgré sa position arrière-arc, aucune contamination géochimique caractéristique d'une subduction n'est perceptible. L'échantillonnage étudié montre que les réservoirs magmatiques de ce segment en propagation évoluent dans une perpétuelle dynamique de recherche d'équilibre thermique et chimique, perturbée par les actions conjuguées suivantes : l'éloignement de la pointe du propagateur par rapport aux zones de réalimentations focalisées sous le centre du segment, des réalimentations successives par des liquides primitifs évoluant avec l'état de maturité du propagateur, et un effet de paroi froide provenant du segment N15, dans lequel la lithosphère de la pointe du segment NS se propage. (Résumé d'auteur

Coupling of adenovirus to transferrin-polylysine/DNA complexes greatly enhances receptor-mediated gene delivery and expression of transfected genes.

We are developing efficient methods for gene transfer into tissue culture cells. We have previously shown that coupling of a chimeric adenovirus with polylysine allowed the construction of an adenovirus-polylysine-reporter-gene complex that transferred the transporter gene with great efficiency into HeLa cells. We have now explored simpler, biochemical means for coupling adenovirus to DNA/polylysine complexes and show that such complexes yield virtually 100% transfection in tissue culture cell lines. In these methods adenovirus is coupled to polylysine, either enzymatically through the action of transglutaminase or biochemically by biotinylating adenovirus and streptavidinylating the polylysine moiety. Combination complexes containing DNA, adenovirus-polylysine, and transferrin-polylysine have the capacity to transfer the reporter gene into adenovirus-receptor- and/or transferrin-receptor-rich cells

Unconfined Specimens

ABSTRACT: Effective diffusion coefficients, D*, of chloride and zinc diffusing in saturated, unconfined specimens of a compacted sandclay mixture are measured for three specimen lengths, L (2.91, 5.83, and 11.60 cm) and three test durations (7, 14, and 21 days). For a specimen length of 2.91 cm, both the chloride and zinc D* values tend to decrease with increasing test duration, possibly due to the measurement of concentration-dependent D* values. For a 14-day test duration, no consistent trend in D* with specimen length is observed, but the overall effect of specimen length on D* is minor relative to the range of measured D* values. A 21-day test duration provides the best correlation between the D* values based on reservoir concentrations, DiScs, and the D* values based on soil concentrations, D~oil, for chloride for a given test regardless of the specimen length. The effect of test duration on the correlation between D~cs and D~oit for zinc is minor based on the relatively narrow range of measured zinc D* values. The observed effects of specimen length on the correlation between D~¢s and D~oil for a given test are consistent with the more uniform final porosity distributions in the shorter specimens and the contrasting effects of the non-linear distributions in porosity and dry density that become less significant as the specimen length increases. KEYWORDS: adsorption, attapulgite clay, batch equilibrium, chloride diffusion, contaminant transport, diffusion testing, Freundlich isotherm, sand-clay mixture, swelling, zinc diffusion Over the past -30 years, diffusion testing has been performed in several different disciplines (e.g., soil science, geology, oceanography, geotechnical engineering) for several different purposes, including diffusion of nutrients to plant roots (Olsen and Kemper 1968), characterization of pore water in geologic deposits Several different types of diffusion testing procedures can be used, test durations have ranged from a few hours to several months, and the specimen volumes have ranged from as small as 10 cm 3 to more than 944 cm 3 (Shacketford 1991). Practical limitations to some, if not all, of the different test methods undoubtedly exist, and variability in test duration and specimen size may have an effect on determination of the measured diffusion coefficients. As a result, an evaluation of the factors potentially affecting the measurement of effective diffusion coefficients is needed. Such an evaluation is particularly of interest to geotechnical engineers associated with the design and evaluation of waste containment barriers because of the increasing importance placed on contaminant transport, in general, and diffusive transport, in particular, in such applications. The primary objective of this study is to evaluate the potential influence of test duration and specimen length on the diffusion of chloride and zinc in compacted, unconfined specimens of a sandclay mixture. The evaluation is based on the single reservoir, decreasing source concentration method that has been used extensively in the measurement of effective diffusion coefficients associated with waste disposal applications Materials and Methods Soil The soil used in this study is a mixture of 75% sand and 25% attapulgite clay (dry weight basis). Physical and chemical properties of the sand and attapulgite clay are provided i

Using reference-free compressed data structures to analyze sequencing reads from thousands of human genomes.

We are rapidly approaching the point where we have sequenced millions of human genomes. There is a pressing need for new data structures to store raw sequencing data and efficient algorithms for population scale analysis. Current reference-based data formats do not fully exploit the redundancy in population sequencing nor take advantage of shared genetic variation. In recent years, the Burrows-Wheeler transform (BWT) and FM-index have been widely employed as a full-text searchable index for read alignment and de novo assembly. We introduce the concept of a population BWT and use it to store and index the sequencing reads of 2705 samples from the 1000 Genomes Project. A key feature is that, as more genomes are added, identical read sequences are increasingly observed, and compression becomes more efficient. We assess the support in the 1000 Genomes read data for every base position of two human reference assembly versions, identifying that 3.2 Mbp with population support was lost in the transition from GRCh37 with 13.7 Mbp added to GRCh38. We show that the vast majority of variant alleles can be uniquely described by overlapping 31-mers and show how rapid and accurate SNP and indel genotyping can be carried out across the genomes in the population BWT. We use the population BWT to carry out nonreference queries to search for the presence of all known viral genomes and discover human T-lymphotropic virus 1 integrations in six samples in a recognized epidemiological distribution

Alternate primers for whole-genome SARS-CoV-2 sequencing.

As the world is struggling to control the novel Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), there is an urgency to develop effective control measures. Essential information is encoded in the virus genome sequence with accurate and complete SARS-CoV-2 sequences essential for tracking the movement and evolution of the virus and for guiding efforts to develop vaccines and antiviral drugs. While there is unprecedented SARS-CoV-2 sequencing efforts globally, approximately 19 to 43 per cent of the genomes generated monthly are gapped, reducing their information content. The current study documents the genome gap frequencies and their positions in the currently available data and provides an alternative primer set and a sequencing scheme to help improve the quality and coverage of the genomes