Top Tips for Designing and Managing a Public Engagement Laboratory.

Public engagement with science is a core facet of the broader science ecosystem, in particular the science research and science education sectors. In this article we demarcate the benefits of dedicated laboratories along with practitioner advice pertaining to the design and running of a public engagement learning environment. A practicing public engagement laboratory and one that is currently being developed are used as illustrative cases to provide real-world insights to public engagement practitioners.

The rhesus incompatible pregnancy and its consequences for affected fetuses and neonates.

Rhesus incompatibility in pregnancy may result in haemolytic disease of the fetus and newborn (HDFN). This review discusses the fetal, neonatal and long-term consequences of HDFN and its management. Untreated, the fetal and neonatal prognosis of HDFN is poor.

[Therapeutic intensification and autologous stem cell transplantation in autoimmune diseases].

THE PATHOPHYSIOLOGY of most autoimmune diseases is often poorly understood. EXPERIMENTAL CONSIDERATIONS and clinical experience suggest that high doses immunoablation followed by stem cell transplantation is a therapeutic option to consider for certain severe autoimmune disorders. THE CONCEPT OF RESTORING NORMAL IMMUNE REACTIVITY must in part br true since current results of 466 transplants (445 autologous, 21 allogeneic) patients suffering from various autoimmune diseases show a beneficial outcome in approximately 2/3 of the patients.

SHV-34: an extended-spectrum beta-lactamase encoded by an epidemic plasmid.

Objectives: To elucidate the causes for treatment failure in children given extended-spectrum cephalosporins.

Methods: During April 1998-March 2000, 18 isolates of members of the family Enterobacteriaceae, fulfilling microbiological criteria for carriage of extended-spectrum beta-lactamases (ESBLs) and carrying blaSHV, were isolated from paediatric inpatients. The collection was subjected to a retrospective molecular analysis.

The type IC hsd loci of the enterobacteria are flanked by DNA with high homology to the phage P1 genome: implications for the evolution and spread of DNA restriction systems.

EcoR124l, EcoDXXl and Ecoprrl are the known members of the type IC family of DNA restriction and modification systems. The first three are carried on large, conjugative plasmids, while Ecoprrl is chromosomally encoded. The enzymes are coded by three genes, hsdR, hsdM and hsdS.

The Escherichia coli prr region encodes a functional type IC DNA restriction system closely integrated with an anticodon nuclease gene.

The prr locus was originally described as coding a ribonuclease that is activated after phage T4 infection to cut within the anticodon of a specific tRNA, inactivating protein synthesis and thus blocking phage development. Wild-type T4 phage has two genes coding the enzymes polynucleotide kinase and RNA ligase, whose only function seems to be to repair the damage done by the anticodon nuclease. As the only apparent function of the prr ribonuclease is to combat phage infection, it can be considered as an RNA-based restriction enzyme.

Binding of proteins from embryonic and differentiated cells to a bidirectional promoter contained within a CpG island.

We have analysed binding sites of nuclear protein factors to a CpG island (HTF9), which contains the promoter for a pair of overlapping, divergently-transcribed "housekeeping" genes. Using DNaseI protection assays with extracts from a range of differentiated and undifferentiated cell lines, including mouse embryonic stem (ES) and embryonal carcinoma (EC) cells, we located multiple protein binding sites on HTF9. Most of the sites were outside the defined core promoter and could bind to previously identified transcription factors.

Location of sequences within rotavirus SA11 glycoprotein VP7 which direct it to the endoplasmic reticulum.

The Simian 11 rotavirus glycoprotein VP7 is directed to the endoplasmic reticulum (ER) of the cell and retained as an integral membrane protein. The gene coding for VP7 predicts two potential initiation codons, each of which precedes a hydrophobic region of amino acids (H1 and H2) with the characteristics of a signal peptide. Using the techniques of gene mutagenesis and expression, we have determined that either hydrophobic domain alone can direct VP7 to the ER.

Ovarian follicular maturation in women. I. Intermittent versus daily administration of human menopausal gonadotropin.

This study was designed to compare the efficiency of daily intramuscular human menopausal gonadotropin (hMG) therapy to an intermittent subcutaneous regimen in women with ovulatory dysfunction. Ovarian follicular development was assessed with the use of serial ultrasound scans and serum 17 beta-estradiol (E2) levels. Ovulation was based on pregnancy and/or elevated luteal progesterone values.

Deletions into an NH2-terminal hydrophobic domain result in secretion of rotavirus VP7, a resident endoplasmic reticulum membrane glycoprotein.

Rotavirus, a non-enveloped reovirus, buds into the rough endoplasmic reticulum and transiently acquires a membrane. The structural glycoprotein, VP7, a 38-kD integral membrane protein of the endoplasmic reticulum (ER), presumably transfers to virus in this process. The gene for VP7 potentially encodes a protein of 326 amino acids which has two tandem hydrophobic domains at the NH2-terminal, each preceded by an in-frame ATG codon.

Polyoma virus DNA replication requires an enhancer.

Sequences which activate polyoma virus DNA replication are located within a region that also includes the transcriptional enhancer. We demonstrate a cis involvement of enhancers in DNA replication by showing that this region can be replaced by other enhancers, in a position- and orientation-independent manner, and that an immunoglobulin gene enhancer confers tissue-specific replicatory ability.

Deletion mapping of a short polyoma virus middle T antigen segment important for transformation.

Viable polyoma virus mutants were constructed that had small deletions in the early region of the genome. The deletions together removed most of the segment missing from the genome of the nontransforming mutant dl23 (N. Smolar and B.

Transcriptional 'enhancers' from SV40 and polyoma virus show a cell type preference.

Transcriptional "enhancers" have been identified in both the monkey virus SV40 and in the mouse polyoma virus. Here we report that these enhancers show a cell type preference. This was done, (i) by assaying for T antigen expression and viral DNA replication of polyoma DNA which contains its own enhancer or the enhancer of SV40 virus, and (ii) by linking either of the two enhancer elements to the rabbit beta 1-globin gene and measuring transient globin expression in mouse and primate cells.

DNA sequences required for specific and efficient initiation of transcription at the polyoma virus early promoter.

The 5'-flanking DNA sequences involved in the specific and efficient transcription of the polyoma virus early region have been investigated. Sequence requirements for efficient in vivo expression differed from those in vitro. Deletion of DNA located between 200 and 400 base pairs before the principal cap sites severely inhibited in vivo expression as measured by transformation ability, but did not affect in vitro transcription.

Sequences at the capped 5'-ends of polyoma virus late region mRNAs: an example of extreme terminal heterogeneity.

We have localized with respect to the genomic DNA sequence the capped 5'-termini of polyoma virus late region mRNAs. A minimum of fifteen different purine termini were found within a 94 base pair region (66.36 to 68.

A region of the polyoma virus genome between the replication origin and late protein coding sequences is required in cis for both early gene expression and viral DNA replication.

Deletion mutants within the Py DNA region between the replication origin and the beginning of late protein coding sequences have been constructed and analysed for viability, early gene expression and viral DNA replication. Assay of replicative competence was facilitated by the use of Py transformed mouse cells (COP lines) which express functional large T-protein but contain no free viral DNA. Viable mutants defined three new nonessential regions of the genome.

The high affinity binding site on polyoma virus DNA for the viral large-T protein.

In order to map the high affinity binding site for the viral large-T protein on polyoma virus DNA, we have developed an assay which does not require purified protein. It is based on the specific elution of the large-T ATPase activity from calf thymus DNA cellulose by recombinant DNA molecules including known sequences of the viral DNA. Using this assay, a high affinity binding site has been mapped on the early region side of the ori region.

Replication and interaction of virus DNA and cellular DNA in mouse cells infected by a human adenovirus.

C57 black mouse cells infected with human adenovirus type 5 (Ad5) produced large amounts of early virus proteins, small amounts of late virus proteins and less than 0.2 infectious units (i.u.

Some adenovirus DNA is associated with the DNA of permissive cells during productive or restricted growth.

We have investigated the association of viral DNA with cell DNA in chicken embryo kidney (CEK) cells productively infected with chicken embryo lethal orphan (CELO) virus and in human (HEK) cells infected with mutants ts36 and ts125 of human adenovirus type 5 under permissive and restrictive conditions. Cell and viral DNA molecules were separated after CELO virus infection of CEK cells by alkaline sucrose gradient centrifugation, network formation, and CsCl density gradient centrifugation, methods that rely on different properties of the DNA. The cell DNA was then tested for viral sequences by DNA reannealing kinetics.