Neurally adjusted ventilatory assist for children on veno-venous ECMO.

NAVA may improve veno-venous ECMO weaning in children. This is a retrospective small series, describing for the first time proof-of-principle for the use of NAVA in children on VV ECMO. Six patients (age 1-48 months) needed veno-venous ECMO.

Divergent prion strain evolution driven by PrP expression level in transgenic mice.

Prions induce a fatal neurodegenerative disease in infected host brain based on the refolding and aggregation of the host-encoded prion protein PrP into PrP. Structurally distinct PrP conformers can give rise to multiple prion strains. Constrained interactions between PrP and different PrP strains can in turn lead to certain PrP (sub)populations being selected for cross-species transmission, or even produce mutation-like events.

Diclofenac potassium liquid-filled soft gelatin capsules in the management of patients with postbunionectomy pain: a Phase III, multicenter, randomized, double-blind, placebo-controlled study conducted over 5 days.

Background: Diclofenac potassium liquid-filled soft gelatin capsule (DPSGC) is a rapidly absorbed formulation of diclofenac potassium developed for the treatment of mild to moderate acute pain.

Objective: The present study was conducted to assess the efficacy and safety profile of DPSGC 25 mg in patients with pain after first-metatarsal bunionectomy.

Methods: This was a Phase III, multicenter, randomized, double-blind, parallel-group, placebo-controlled study conducted over 5 days.

Distal element(s) is(are) required for position-independent expression of the goat alpha-lactalbumin gene in transgenic mice. Potential relationship with the location of the cyclin T1 locus.

We recently reported the site-independent and copy-number-related expression in mice of a goat alpha-lactalbumin gene with 150 kb and 10 kb of 5'- and 3'-flanking sequences, respectively. In the present study, we observed that the resection of the 5'-flanking region, leaving only 70 kb, resulted in a site-dependent expression of this milk protein-encoding transgene. This suggests that important cis-regulatory elements are located within the distal-deleted sequence.

Heterogeneous inducible mammary-specific expression of Jab/SOCS1 in lactating transgenic mice is associated with no obvious phenotype, even at the cellular level.

The cytokine-inducible suppressor of cytokine signalling SOCS1, or JAB, has been shown to be implicated in vitro in the negative regulation of the prolactin-receptor-induced activation of JAK2 and STAT5. Disruption of this gene in vivo resulted in an accelerated mammary gland development. In the present experiment, we assessed the potential impact on the lactation process of the doxycycline-inducible mammary-controlled expression of this gene in transgenic mice.

Substitution of the alpha-lactalbumin transcription unit by a CAT cDNA within a BAC clone silenced the locus in transgenic mice without affecting the physically linked Cyclin T1 gene.

We recently reported that a goat bacterial artificial chromosome (BAC) clone conferred site-independent expression in transgenic mice of the two loci present within its insert, the ubiquitously expressed Cyclin T1 and the mammary specific alpha-lactalbumin (alphalac) genes. To assess if this vector could target mammary-restricted expression of cDNA, the CAT ORF was introduced by homologous recombination in Escherichia coli in place of the alphalac transcription unit. The insert of this modified BAC was injected into mice and three transgenic lines were derived.

New in vivo and ex vivo models for the experimental study of sheep scrapie: development and perspectives.

Sheep scrapie is a prototypical transmissible spongiform encephalopathy (TSE), and the most widespread of these diseases. Experimental study of TSE infectious agents from sheep and other species essentially depends on bioassays in rodents. Transmission of natural sheep scrapie to conventional mice commonly requires one or two years.

Markedly increased susceptibility to natural sheep scrapie of transgenic mice expressing ovine prp.

The susceptibility of sheep to scrapie is known to involve, as a major determinant, the nature of the prion protein (PrP) allele, with the VRQ allele conferring the highest susceptibility to the disease. Transgenic mice expressing in their brains three different ovine PrP(VRQ)-encoding transgenes under an endogenous PrP-deficient genetic background were established. Nine transgenic (tgOv) lines were selected and challenged with two scrapie field isolates derived from VRQ-homozygous affected sheep.

Introduction of a proximal Stat5 site in the murine alpha-lactalbumin promoter induces prolactin dependency in vitro and improves expression frequency in vivo.

In order to establish a possible correlation between in vitro prolactin induction and the transcriptional activity of mammary gene promoters in transgenic mice, a functional Stat5-binding site was created by means of site-directed mutagenesis at position -70 on a 560 bp murine alpha-lactalbumin promotor linked to a CAT reporter gene. Surprisingly, the wild-type promoter was constitutively active in vitro and could not be induced by prolactin. Introducing the proximal Stat5 site abolished this constitutive activity and resulted in prolactin dependence in both CHO-K1- and HC11-transfected cells.

Use of doxycycline-controlled gene expression to reversibly alter milk-protein composition in transgenic mice.

A reverse tetracycline transactivator-encoding cDNA under the control of the mammary specific beta-lactoglobulin promoter was linked to a bovine alpha-lactalbumin transcription unit driven by a reverse tetracycline-controlled transactivator/doxycycline-inducible human cytomegalovirus promoter. The construct was microinjected into eggs from alpha-lactalbumin-deficient mice. These mice produce a highly viscous lactose-free milk and have a shortened lactation period.

Position-independent and copy-number-related expression of a goat bacterial artificial chromosome alpha-lactalbumin gene in transgenic mice.

A bacterial artificial chromosome goat insert comprising the alpha-lactalbumin-encoding transcription unit with approximately 150 and 10 kb of 5'- and 3'-flanking sequences, respectively, was micro-injected into mouse eggs. In six out of seven transgenic lines, the level of mammary tissue- and stage-specific expression was position-independent and copy-number-dependent. The exogenous alpha-lactalbumin yield, about 0.

Sequence of murine CDC10 cDNA, gene organization and expression analysis.

The sequence of the 2391 bp murine CDC10 cDNA is reported. The gene transcription unit, composed of 13 exons, encodes a potential 417/446 amino acid GTP-binding protein, highly similar to human CDC10. The ubiquitous expression of the gene, as judged by Northern analysis, is consistent with its putative promoter structure.

The efficacy of ANSAID (flurbiprofen) as an analgesic in foot surgery.

Eighty-three patients scheduled for outpatient surgery were enrolled in this double-blind, randomized trial designed to compare the safety and efficacy of single and multiple doses of flurbiprofen 50 mg. and acetaminophen 300 mg. plus codeine phosphate 30 mg.

Developmental regulation of murine integrin beta 1 subunit- and Hsc73-encoding genes in mammary gland: sequence of a new mouse Hsc73 cDNA.

A partial integrin beta 1 subunit-encoding cDNA (Itg beta 1) and a new heat-shock protein 70-like-encoding cDNA (Hsc73) homologous to rat Hsc73 were cloned by differential display and RT-PCR from mouse mammary gland. Their developmental regulation during pregnancy, lactation and involution is reported. The Itg beta 1 mRNA content was stable in the first half of gestation, decreased to a minimum during lactation and increased markedly in early involution.

Efficient and specific ribozyme-mediated reduction of bovine alpha-lactalbumin expression in double transgenic mice.

Transgenic mice carrying a bovine alpha-lactalbumin (alpha-lac) specific ribozyme gene under the transcriptional control of the mouse mammary tumor virus long terminal repeat were generated and cross-bred with animals that highly express a bovine alpha-lac transgene (0.4 mg of alpha-lac/ml(-1) of milk). The ribozyme contains the hammerhead catalytic domain, flanked by 12-nt sequences complementary to the 3' untranslated region of bovine alpha-lac transcript.

Creation and phenotypic analysis of alpha-lactalbumin-deficient mice.

alpha-Lactalbumin is an abundant milk-specific calcium metalloprotein which has an evolutionary relationship to lysozyme. It modifies the substrate specificity of a Golgi galactosyltransferase by forming the lactose synthetase binary complex. Lactose, together with other sugars and diffusible ions, is responsible for the osmotic pressure of milk.

Complete sequence of a bovine alpha-lactalbumin pseudogene: the region homologous to the gene is flanked by two directly repeated LINE sequences.

A 3-kb fragment hybridizing with a bovine alpha-lactalbumin cDNA probe was isolated from a bovine genomic library and completely sequenced. An internal fragment beginning downstream from exon 2, as already reported for another pseudogene, but ending in the 3'-untranslated region of exon 4 shares 78% sequence similarity with the bovine alpha-lactalbumin gene. This region is flanked by two directly repeated LINE sequences.

Isolation and characterization of the mouse alpha-lactalbumin-encoding gene: interspecies comparison, tissue- and stage-specific expression.

The murine alpha-lactalbumin-encoding gene (m alpha La) was isolated and completely sequenced. The 2.3-kb transcription unit shared a similar organization with that of its counterparts from other species.

Sequence of the murine alpha-lactalbumin-encoding cDNA: interspecies comparison of the coding frame and deduced pre-protein.

The structure of the mouse alpha-lactalbumin-encoding mRNA was deduced from sequence analysis of eight cDNA clones. The almost full-length mRNA of 732 nucleotides [poly(A) tail excluded] and the deduced pre-protein share 85% and 86% homology with their rat counterpart, respectively. Interspecies comparison of the pre-protein showed the occurrence of an extra amino acid (aa) in the signal peptide and of two mutations affecting two reported invariant aa residues at positions 44 and 107, which weakens the assumption that both aa residues might play a significant structural and/or functional role.

Expression analysis of ruminant alpha-lactalbumin in transgenic mice: developmental regulation and general location of important cis-regulatory elements.

The bovine alpha-lactalbumin transgene with 750 bp and 336 bp of the 5' and 3' flanking region, respectively, is developmentally regulated as its endogenous counterpart in transgenic mice. Comparative expression analysis of three 5'-shortened constructs suggests that the region -477/-220 contains important cis-acting transcriptional elements. The level of expression of a long caprine alpha-lactalbumin transgene encompassing 8.

The bovine alpha-lactalbumin promoter directs expression of ovine trophoblast interferon in the mammary gland of transgenic mice.

A hybrid construct derived from ovine trophoblastin cDNA and bovine alpha-lactalbumin-encoding gene, was injected into the pronuclei of mouse eggs. In one of the resulting transgenic mouse lines, expression of the hybrid construct was detected and found to be limited to the mammary gland of lactating females which secreted active ovine trophoblastin. This strongly suggests that important cis-acting DNA sequences involved in tissue-specific expression of the bovine gene are located within the second half of the 3' untranslated region, or/and the proximal 5' and 3' regions flanking the transcriptional unit.

Sequence of the goat alpha-lactalbumin-encoding gene: comparison with the bovine gene and evidence of related sequences in the goat genome.

The isolation and characterization of a genomic clone encoding goat alpha-lactalbumin (alpha LA) are described, and its sequence was aligned with that of its bovine counterpart. The 5'-flanking regions are highly homologous, but two small insertions were found in the goat sequence. One of these insertions fills the gap found in the so-called 'milk-box' consensus sequence of the bovine gene.

Efficient tissue-specific expression of bovine alpha-lactalbumin in transgenic mice.

Transgenic mice carrying a 3.2-kb genomic fragment comprising the bovine alpha-lactalbumin gene transcription unit with 750 bp and 336 bp of 5' and 3' flanking regions, respectively, were generated. Tissue-specific expression of the transgene was observed in five out of the six lines analyzed at levels close to the endogenous gene in two of them.