Regulation of the Biomedical Applications of Recombinant DNA Research

In recent years, the rapid expansion of knowledge in the field of molecular genetics resulting from the use of recombinant DNA (rDNA) techniques has been unprecedented. The expanded knowledge scientists have acquired through rDNA techniques has precipitated conspicuous breakthroughs in biomedical research involving the manipulation of human genetic material to diagnose and treat human disorders. Application of this research may soon affect all aspects of our lives. However, this newly-acquired ability to manipulate human genes raises broad ethical and legal questions. The issues raised by rDNA research are dissimilar to earlier questions regarding the use of genetically-engineered microorganisms in the laboratory and current questions related to the regulation of biotechnology. Despite this dissimilarity, the rapidity with which biomedical developments have been achieved makes the resolution of these ethical and legal questions regarding rDNA techniques all the more urgent

A placebo-controlled trial of folic acid and betaine in identical twins with Angelman syndrome.

BackgroundAngelman syndrome (AS) is a neurodevelopmental disorder that is caused by maternal genetic deficiency of a gene that encodes E6-AP ubiquitin-protein ligase (gene symbol UBE3A) mapping to chromosome 15q11-q13. AS leads to stiff and jerky gait, excess laughter, seizures, and severe intellectual disability. In some parts of the brain, the paternally inherited UBE3A gene is subject to genomic imprinting by the action of the UBE3A-antisense transcript (UBE3A-ATS) on the paternally inherited allele. Consequently, only the maternally inherited UBE3A gene is expressed in mature neurons. AS occurs due to deletions of the maternal 15q11 - 13 region, paternal uniparental disomy (UPD), imprinting center defects, mutations in the maternal UBE3A gene, or other unknown genetic malfunctions that result in a silenced maternal UBE3A gene in the specific imprinted regions of the brain.ResultsA potential treatment strategy for AS is to increase methylation of UBE3A-ATS to promote expression of the paternal UBE3A gene and thus ameliorate the clinical phenotypes of AS. We treated two sets of male identical twins with class I deletions with a 1 year treatment trial of either betaine and folic acid versus placebo. We found no statistically significant changes in the clinical parameters tested at the end of the 1 year trial, nor did we find any significant adverse events.ConclusionsThis study tested the hypothesis that by increasing the methylation of the UBE3A-antisense transcript in Angelman syndrome to promote expression of the silenced paternal UBE3A gene we may ameliorate the clinical phenotypes of AS. We treated two sets of identical twins with placebo versus betaine and folic acid. Although this study represented a novel approach to treating Angelman syndrome, the differences in the developmental testing results was not significant. This paper also discusses the value of monozygotic twin studies in minimizing confounding variables and its utility in conducting small treatment studies.Trial registrationNCT00348933 . Registered 6 July 2006

How the liver keeps itself in shape

After fasting, hepatocytes proliferate to help the liver grow back to its original size

Using physical barriers to prevent carrot fly (Psila rosae (Fabricius)) damage in domestic production

A field experiment was used to assess the effectiveness of different barriers in protecting garden‐scale carrot production from carrot fly (Psila rosae (Fabricius)) damage. Some of the vertical barriers tested were found to provide a useful method of protecting early season carrots from carrot fly in terms of the percentage of carrots free from damage but, under cumulative pest pressure of several generations of carrot fly, such barriers were found to provide insufficient protection. Gardeners should therefore completely cover their carrot crop to attain an acceptable level of control, and this was found to be especially important for carrots harvested later in the season. There were positive effects of some barrier types on yield which may be due, at least in part, to the protection given by the barriers to carrot seedlings

Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice

Mice are a widely used pre-clinical model system in large part due to their potential for genetic manipulation. The ability to manipulate gene expression in specific cells under temporal control is a powerful experimental tool. The liver is central to metabolic homeostasis and a site of many diseases, making the targeting of hepatocytes attractive. Adeno-associated virus 8 (AAV8) vectors are valuable instruments for the manipulation of hepatocellular gene expression. However, their off-target effects in mice have not been thoroughly explored. Here, we sought to identify the short-term off-target effects of AAV8 administration in mice. To do this, we injected C57BL/6J wild-type mice with either recombinant AAV8 vectors expressing Cre recombinase or control AAV8 vectors and characterised the changes in general health and in liver physiology, histology and transcriptomics compared to uninjected controls. We observed an acute and transient trend for reduction in homeostatic liver proliferation together with induction of the DNA damage marker γH2AX following AAV8 administration. The latter was enhanced upon Cre recombinase expression by the vector. Furthermore, we observed transcriptional changes in genes involved in circadian rhythm and response to infection. Notably, there were no additional transcriptomic changes upon expression of Cre recombinase by the AAV8 vector. Overall, there was no evidence of liver injury, and only mild T-cell infiltration was observed 14 days following AAV8 infection. These data advance the technique of hepatocellular genome editing through Cre-Lox recombination using Cre expressing AAV vectors, demonstrating their minimal effects on murine physiology and highlight the more subtle off target effects of these systems

Turnover in floral composition explains species diversity and temporal stability in the nectar supply of urban residential gardens

Residential gardens are a valuable habitat for insect pollinators worldwide, but differences in individual gardening practices substantially affect their floral composition. It is important to understand how the floral resource supply of gardens varies in both space and time so we can develop evidence‐based management recommendations to support pollinator conservation in towns and cities. We surveyed 59 residential gardens in the city of Bristol, UK, at monthly intervals from March to October. For each of 472 garden surveys, we combined floral abundances with nectar sugar data to quantify the nectar production of each garden, investigating the magnitude, temporal stability, and diversity and composition of garden nectar supplies. We found that individual gardens differ markedly in the quantity of nectar sugar they supply (from 2 to 1,662 g), and nectar production is higher in more affluent neighbourhoods, but not in larger gardens. Nectar supply peaks in July (mid‐summer), when more plant taxa are in flower, but temporal patterns vary among individual gardens. At larger spatial scales, temporal variability averages out through the portfolio effect, meaning insect pollinators foraging across many gardens in urban landscapes have access to a relatively stable and continuous supply of nectar through the year. Turnover in species composition among gardens leads to an extremely high overall plant richness, with 636 taxa recorded flowering. The nectar supply is dominated by non‐natives, which provide 91% of all nectar sugar, while shrubs are the main plant life form contributing to nectar production (58%). Two‐thirds of nectar sugar is only available to relatively specialised pollinators, leaving just one‐third that is accessible to all. Synthesis and applications. By measuring nectar supply in residential gardens, our study demonstrates that pollinator‐friendly management, affecting garden quality, is more important than the size of a garden, giving every gardener an opportunity to contribute to pollinator conservation in urban areas. For gardeners interested in increasing the value of their land to foraging pollinators, we recommend planting nectar‐rich shrubs with complementary flowering periods and prioritising flowers with an open structure in late summer and autumn

Ploidy dynamics increase the risk of liver cancer initiation

Liver cancer typically arises after years of inflammatory insults to hepatocytes. These cells can change their ploidy state during health and disease. Whilst polyploidy may offer some protection, new research shows it may also promote the formation of liver tumours

The low redshift Lyman-α\alpha Forest as a constraint for models of AGN feedback

We study the low redshift Lyman-$\alpha$ Forest in the Illustris and IllustrisTNG (TNG) cosmological simulations to demonstrate their utility in constraining aspects of sub-grid models of feedback from active galactic nuclei (AGN). The two simulations share an identical Ultraviolet Background prescription and similar cosmological parameters, but TNG features an entirely reworked AGN feedback model. Therefore a comparison of these simulations is useful to assess the effects of an altered AGN sub-grid model on the low redshift Lyman-$\alpha$ Forest. We find significant differences in the IGM temperature-density relation between the two simulations due to changes in the gas heating rate due to AGN. We investigate Lyman-$\alpha$ Forest observables such as the column density distribution function, flux PDF, and Doppler width ($b$-parameter) distribution. Due to the AGN radio mode model, the original Illustris simulations have a factor of 2-3 fewer absorbers than TNG at column densities $N_{\rm HI}< 10^{15.5}$ cm$^{-2}$. We show that TNG is in much better agreement with the observed $z=0.1$ flux power spectrum than Illustris. The differences in the amplitude and shape of the flux PDF and power spectrum between Illustris and TNG cannot be attributed to simple changes in the photoheating rate. We also compare the simulated Forest statistics to UV data from the Cosmic Origins Spectrograph (COS) and find that neither simulation can reproduce the slope of the absorber distribution. Both Illustris and TNG also produce significantly smaller $b$-parameter distributions than observed in the COS data, possibly due to unresolved or missing sources of turbulence.Comment: Submitted to ApJL, comments welcom

Human hippocampal theta power indicates movement onset and distance travelled

Theta frequency oscillations in the 6- to 10-Hz range dominate the rodent hippocampal local field potential during translational movement, suggesting that theta encodes self-motion. Increases in theta power have also been identified in the human hippocampus during both real and virtual movement but appear as transient bursts in distinct high- and low-frequency bands, and it is not yet clear how these bursts relate to the sustained oscillation observed in rodents. Here, we examine depth electrode recordings from the temporal lobe of 13 presurgical epilepsy patients performing a selfpaced spatial memory task in a virtual environment. In contrast to previous studies, we focus on movement-onset periods that incorporate both initial acceleration and an immediately preceding stationary interval associated with prominent theta oscillations in the rodent hippocampal formation. We demonstrate that movementonset periods are associated with a significant increase in both low (2–5 Hz)- and high (6–9 Hz)-frequency theta power in the human hippocampus. Similar increases in low- and high-frequency theta power are seen across lateral temporal lobe recording sites and persist throughout the remainder of movement in both regions. In addition, we show that movement-related theta power is greater both before and during longer paths, directly implicating human hippocampal theta in the encoding of translational movement. These findings strengthen the connection between studies of theta-band activity in rodents and humans and offer additional insight into the neural mechanisms of spatial navigation