Subunit recombinant vaccine protects against monkeypox

The smallpox vaccine Dryvax, a live vaccinia virus (VACV), protects against smallpox and monkeypox, but is contraindicated in immunocompromised individuals. Because Abs to VACV mediate protection, a live virus vaccine could be substituted by a safe subunit protein-based vaccine able to induce a protective Ab response. We immunized rhesus macaques with plasmid DNA encoding the monkeypox orthologs of the VACV L1R, A27L, A33R, and B5R proteins by the intradermal and i.m. routes, either alone or in combination with the equivalent recombinant proteins produced in Escherichia coli. Animals that received only DNA failed to produce high titer Abs, developed innumerable skin lesions after challenge, and died in a manner similar to placebo controls. By contrast, the animals vaccinated with proteins developed moderate to severe disease (20-155 skin lesions) but survived. Importantly, those immunized with DNA and boosted with proteins had mild disease with 15 or fewer lesions that resolved within days. DNA/protein immunization elicited Th responses and binding Ab titers to all four proteins that correlated negatively with the total lesion number. The sera of the immunized macaques recognized a limited number of linear B cell epitopes that are highly conserved among orthopoxviruses. Their identification may guide future efforts to develop simpler, safer, and more effective vaccines for monkeypox and smallpox.     

The establishment and regeneration of a range ofCuphea germplasm t in vitro

The range of Cuphea species previously investigated in vitro has been extended, and the potential of a number of species for uptake into gene transfer systems evaluated. Eight species were successfully established in culture and assessments of relative subculturing potential rates after 4 weeks varied from 10.3 per initial shoot in C. wrightii to 2.3 in C. leptopoda. In tissue regeneration studies, callus formation was rapid and efficient from seven of the eight lines investigated and the species C. lutea, C. lanceolata, C. leptopoda, C. paucipetala and C. laminuligera (in addition to the previously reported C. wrightii) regenerated shootsin vitro from leaf explants after three to six weeks culture on a medium containing 8.8 μM BA, in a two-stage protocol. A high incidence of in vitro flowering was also recorded in C. lutea, C. leptopoda and C. paucipetala. Though initial growth rates were comparable, it was found that extended culture on a matrix of Phytagel rather than agar was detrimental to subsequent shoot development. This revised version was published online in June 2006 with corrections to the Cover Date.     

Diverse Regulators of Human Ribosome Biogenesis Discovered by Changes in Nucleolar Number

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Incidental Findings on Brain MR Imaging in Older Community-Dwelling Subjects Are Common but Serious Medical Consequences Are Rare: A Cohort Study

Objectives

Incidental findings in neuroimaging occur in 3% of volunteers. Most data come from young subjects. Data on their occurrence in older subjects and their medical, lifestyle and financial consequences are lacking. We determined the prevalence and medical consequences of incidental findings found in community-dwelling older subjects on brain magnetic resonance imaging.

Design

Prospective cohort observational study.

Setting

Single centre study with input from secondary care.

Participants

Lothian Birth Cohort 1936, a study of cognitive ageing.

Main Outcome Measures

Incidental findings identified by two consultant neuroradiologists on structural brain magnetic resonance imaging at age 73 years; resulting medical referrals and interventions.

Primary and Secondary Outcome Measures

Prevalence of incidental findings by individual categories: neoplasms, cysts, vascular lesions, developmental, ear, nose or throat anomalies, by intra- and extracranial location; visual rating of white matter hyperintensities and brain atrophy.

Results

There were 281 incidental findings in 223 (32%) of 700 subjects, including 14 intra- or extracranial neoplasms (2%), 15 intracranial vascular anomalies (2%), and 137 infarcts or haemorrhages (20%). Additionally, 153 had moderate/severe deep white matter hyperintensities (22%) and 176 had cerebral atrophy at, or above, the upper limit of normal (25%) compared with a normative population template. The incidental findings were unrelated to white matter hyperintensities or atrophy; about a third of subjects had both incidental findings and moderate or severe WMH and a quarter had incidental findings and atrophy. The incidental findings resulted in one urgent and nine non-urgent referrals for further medical assessment, but ultimately in no new treatments.

Conclusions

In community-dwelling older subjects, incidental findings, including white matter hyperintensities and atrophy, were common. However, many findings were not of medical importance and, in this age group, most did not result in further assessment and none in change of treatment.     

Molecular interactions between fibronectin and integrins during mouse blastocyst outgrowth

To investigate the mechanism of trophoblast adhesion to fibronectin, we cultured blastocysts in serum-free medium on proteolytic fibronectin fragments containing its major functional domains, and localized fibronectin-binding integrins in outgrowing trophoblast cells by immunofluorescent staining. Outgrowth comparable to that obtained with intact fibronectin was observed using a 120 kD chymotryptic fragment containing the central cell-binding domain (FN-120) and the Arg-Gly-Asp (RGD) recognition sequence. A 40 kD COOH-terminal chymotryptic fragment of fibronectin containing both a heparin-binding region and an alternate (non-RGD) cell-binding site was inactive in supporting trophoblast adhesion. Three synthetic peptides derived from the heparin-binding domain, including the CS1 alternate cell-binding site, were also unable to promote trophoblast cell adhesion. A 75 kD recombinant protein, ProNectin F, containing 13 copies of the cell recognition epitope of fibronectin, Val-Thr-Gly-Arg-Gly-Asp-Ser-Pro-Ala-Ser, vigorously supported blastocyst outgrowth. Blastocyst outgrowth was not significantly different when surfaces were precoated with cellular fibronectin, which contains an alternatively spliced type III repeat and is the form actually encountered in vivo. Several putative fibronectin receptors were localized in trophoblast outgrowths by immunofluorescent labeling. Antibodies reactive with integrin subunits α3, α5, αllb, αv, β1 and β3, but not α4, all bound to trophoblast cells. Antibodies raised against either the β1 or β3 integrin subunits significantly inhibited fibronectin-mediated outgrowth. These findings demonstrate the key role of the central cell-binding domain of fibronectin in trophoblast adhesion, and suggest four RGD-binding integrins, α3β1, α5β1, αllbβ3, and αvβ3, that could mediate trophoblast adhesion in vitro and may play an important role during implantation. © 1995 Wiley-Liss, Inc.     

Variation in clone structure of fragmenting coral reef sponges

Populations of three branching Caribbean demosponge species are composed of clones produced by asexual fragmentation. Dispersal of the fragments before they become established as independent individuals scatters clone members widely and intermixes members of different clones, complicating study of the clone structure of these populations and contrasting with many other sessile clonal organisms. Clone structures of these populations were inferred using a combination of tissue-compatibility relationships and an analysis of variations in morphology and colour. Although tissue compatibility cannot be used for precise identification of sponge clones, in general, patterns of variation in morphological characters influencing fragmentation and patterns of fragment dispersal and recruitment suggest that, in these populations, tissue-compatibility relationships closely reflect clone structure. Conditions that must be met in order to use tissue compatibility for study of sponge clones are discussed, and previous results, from which conflicting conclusions have been drawn, reconciled in this context. Variations among clones in numbers of physiologically independent members and in size and shape of areal extent are discussed in the context of processes that may affect evolution of clonal characters in these populations and in other species that propagate by dispersing asexual fragments.