Assessment of a novel, capsid-modified adenovirus with an improved vascular gene transfer profile

<p>Background: Cardiovascular disorders, including coronary artery bypass graft failure and in-stent restenosis remain significant opportunities for the advancement of novel therapeutics that target neointimal hyperplasia, a characteristic of both pathologies. Gene therapy may provide a successful approach to improve the clinical outcome of these conditions, but would benefit from the development of more efficient vectors for vascular gene delivery. The aim of this study was to assess whether a novel genetically engineered Adenovirus could be utilised to produce enhanced levels of vascular gene expression.</p> <p>Methods: Vascular transduction capacity was assessed in primary human saphenous vein smooth muscle and endothelial cells using vectors expressing the LacZ reporter gene. The therapeutic capacity of the vectors was compared by measuring smooth muscle cell metabolic activity and migration following infection with vectors that over-express the candidate therapeutic gene tissue inhibitor of matrix metalloproteinase-3 (TIMP-3).</p> <p>Results: Compared to Adenovirus serotype 5 (Ad5), the novel vector Ad5T*F35++ demonstrated improved binding and transduction of human vascular cells. Ad5T*F35++ mediated expression of TIMP-3 reduced smooth muscle cell metabolic activity and migration in vitro. We also demonstrated that in human serum samples pre-existing neutralising antibodies to Ad5T*F35++ were less prevalent than Ad5 neutralising antibodies.</p> <p>Conclusions: We have developed a novel vector with improved vascular transduction and improved resistance to human serum neutralisation. This may provide a novel vector platform for human vascular gene transfer.</p&gt

XMM–Newton campaign on ultraluminous X-ray source NGC 1313 X-1: wind versus state variability

Most ultraluminous X-ray sources (ULXs) are thought to be powered by neutron stars and black holes accreting beyond the Eddington limit. If the compact object is a black hole or a neutron star with a magnetic field ≲1012 G, the accretion disc is expected to thicken and launch powerful winds driven by radiation pressure. Evidence of such winds has been found in ULXs through the high-resolution spectrometers onboardXMM–Newton, but several unknowns remain, such as the geometry and launching mechanism of these winds. In order to better understand ULX winds and their link to the accretion regime, we have undertaken a major campaign with XMM–Newton to study the ULX NGC 1313 X-1, which is known to exhibit strong emission and absorption features from a mildly relativistic wind. The new observations show clear changes in the wind with a significantly weakened fast component (0.2c) and the rise of a new wind phase which is cooler and slower (0.06–0.08c). We also detect for the first time variability in the emission lines which indicates an origin within the accretion disc or in the wind. We describe the variability of the wind in the framework of variable super-Eddington accretion rate and discuss a possible geometry for the accretion disc

Chronic Supplementation With a Mitochondrial Antioxidant (MitoQ) Improves Vascular Function in Healthy Older Adults.

UNLABELLED: Excess reactive oxygen species production by mitochondria is a key mechanism of age-related vascular dysfunction. Our laboratory has shown that supplementation with the mitochondrial-targeted antioxidant MitoQ improves vascular endothelial function by reducing mitochondrial reactive oxygen species and ameliorates arterial stiffening in old mice, but the effects in humans are unknown. Here, we sought to translate our preclinical findings to humans and determine the safety and efficacy of MitoQ. Twenty healthy older adults (60-79 years) with impaired endothelial function (brachial artery flow-mediated dilation 7.60 m/s; n=11). Plasma oxidized LDL (low-density lipoprotein), a marker of oxidative stress, also was lower after MitoQ versus placebo (P0.1). These findings in humans extend earlier preclinical observations and suggest that MitoQ and other therapeutic strategies targeting mitochondrial reactive oxygen species may hold promise for treating age-related vascular dysfunction. CLINICAL TRIAL REGISTRATION: URL: http://www.clinicaltrials.gov. Unique identifier: NCT02597023.This work was supported by National Institutes of Health (NIH) awards AG049451, AG000279, AG053009, Colorado CTSA UL1 TR001082, and an industry contract with MitoQ Limited (MitoQ Limited provided MitoQ and some financial support). M.P. Murphy is supported by UK MRC MC_U105663142 and as a Wellcome Trust Investigator (110159/Z/15/Z)

Alcohol-induced retrograde facilitation renders witnesses of crime less suggestible to misinformation

RATIONALE: Research has shown that alcohol can have both detrimental and facilitating effects on memory: intoxication can lead to poor memory for information encoded after alcohol consumption (anterograde amnesia) and may improve memory for information encoded before consumption (retrograde facilitation). This study examined whether alcohol consumed after witnessing a crime can render individuals less vulnerable to misleading post-event information (misinformation). METHOD: Participants watched a simulated crime video. Thereafter, one third of participants expected and received alcohol (alcohol group), one third did not expect but received alcohol (reverse placebo), and one third did not expect nor receive alcohol (control). After alcohol consumption, participants were exposed to misinformation embedded in a written narrative about the crime. The following day, participants completed a cued-recall questionnaire about the event. RESULTS: Control participants were more likely to report misinformation compared to the alcohol and reverse placebo group. CONCLUSION: The findings suggest that we may oversimplify the effect alcohol has on suggestibility and that sometimes alcohol can have beneficial effects on eyewitness memory by protecting against misleading post-event information

A translocation motif in relaxase TrwC specifically affects recruitment by its conjugative type IV secretion system

Type IV secretion system (T4SS) substrates are recruited through a translocation signal that is poorly defined for conjugative relaxases. The relaxase TrwC of plasmid R388 is translocated by its cognate conjugative T4SS, and it can also be translocated by the VirB/D4 T4SS of Bartonella henselae, causing DNA transfer to human cells. In this work, we constructed a series of TrwC variants and assayed them for DNA transfer to bacteria and human cells to compare recruitment requirements by both T4SSs. Comparison with other reported relaxase translocation signals allowed us to determine two putative translocation sequence (TS) motifs, TS1 and TS2. Mutations affecting TS1 drastically affected conjugation frequencies, while mutations affecting either motif had only a mild effect on DNA transfer rates through the VirB/D4 T4SS of B. henselae. These results indicate that a single substrate can be recruited by two different T4SSs through different signals. The C terminus affected DNA transfer rates through both T4SSs tested, but no specific sequence requirement was detected. The addition of a Bartonella intracellular delivery (BID) domain, the translocation signal for the Bartonella VirB/D4 T4SS, increased DNA transfer up to 4% of infected human cells, providing an excellent tool for DNA delivery to specific cell types. We show that the R388 coupling protein TrwB is also required for this high-efficiency TrwC-BID translocation. Other elements apart from the coupling protein may also be involved in substrate recognition by T4SSs

Managerial Work in a Practice-Embodying Institution - The role of calling, the virtue of constancy

What can be learned from a small scale study of managerial work in a highly marginal and under-researched working community? This paper uses the ‘goods-virtues-practices-institutions’ framework to examine the managerial work of owner-directors of traditional circuses. Inspired by MacIntyre’s arguments for the necessity of a narrative understanding of the virtues, interviews explored how British and Irish circus directors accounted for their working lives. A purposive sample was used to select subjects who had owned and managed traditional touring circuses for at least 15 years, a period in which the economic and reputational fortunes of traditional circuses have suffered badly. This sample enabled the research to examine the self-understanding of people who had, at least on the face of it, exhibited the virtue of constancy. The research contributes to our understanding of the role of the virtues in organizations by presenting evidence of an intimate relationship between the virtue of constancy and a ‘calling’ work orientation. This enhances our understanding of the virtues that are required if management is exercised as a domain-related practice

Engineering Mesophase Stability and Structure via Incorporation of Cyclic Terminal Groups

We report on the characterisation of a number of liquid–crystalline materials featuring cyclic terminal groups, which lead to significant enhancements in the temperature range of the mesomorphic state. Materials with only short terminal chains are able to support lamellar mesophase formation by appending a large terminal cyclic unit at the end of a short spacer composed of methylene units. X-ray scattering experiments reveal that the layer spacings of the lamellar smectic phase are significantly larger when a cyclic end-group is present than for equivalent linear unsubstituted materials, but there is no effect on orientational order. Fully atomistic molecular dynamics simulations faithfully reproduce experimental layer spacings and orientational order parameters, and indicate that the cyclic terminal units spontaneously segregate into diffuse sub-layers and thus cause the increased layer spacing. This shape segregation predicted by molecular dynamics simulations is observed in the crystalline solid state by X-ray diffraction

Social representations of AIDS and their quotidian interfaces for people living with HIV

This qualitative descriptive study, guided by the Social Representations Theory, aimed to describe the content of the social representations regarding the Acquired Immunodeficiency Syndrome (AIDS) for seropositive individuals in outpatient monitoring of the public health network and to analyze the interface of the social representations of AIDS with the quotidian of the individuals living with human immunodeficiency virus (HIV), especially in the adherence to treatment process Interviews were conducted with 30 seropositive individuals and the manual content analysis technique was used. From the analysis, six categories emerged that re-translated the quotidian of seropositive people permeated by the stigma, prejudice, struggle for life and the need for the continuous use of antiretrovirals. AIDS was assimilated to chronic diseases such as diabetes, showing a trend of transformation of the social representation of AIDS, substituting the idea of death, with life. It is concluded that people living with HIV are more optimistic due to effective treatments for the control of the disease.Se trata de un estudio cualitativo descriptivo orientado por la Teoría de las Representaciones Sociales, que objetivó describir el contenido de las representaciones sociales acerca de la Síndrome de Inmunodeficiencia Adquirida (SIDA) para los usuarios seropositivos en acompañamiento de ambulatorio en la red pública de salud y analizar la interconexión de las representaciones sociales del Sida con lo cotidiano de los individuos que viven con el virus de la inmunodeficiencia humana (HIV), especialmente al proceso de adhesión al tratamiento. Se realizaron entrevistas con 30 individuos seropositivos. Se utilizó la técnica de análisis de contenido manual. Del análisis, emergieron seis categorías que tradujeron lo cotidiano de seropositivos impregnados por el estigma, prejuicio, lucha por la vida y la necesidad del uso continuo de antirretrovirales. El Sida fue comparado a enfermedades crónicas como la diabetes, evidenciando una tendencia de transformación de la representación social del Sida, substituyendo la idea de muerte, por la de vida. Se concluye que las personas que conviven con HIV están más optimistas debido a los tratamientos eficaces en el control de la enfermedad.Trata-se de estudo qualitativo descritivo, norteado pela teoria das representações sociais. Objetivou-se descrever o conteúdo das representações sociais acerca da síndrome de imunodeficiência adquirida (AIDS) para os usuários soropositivos, em acompanhamento ambulatorial da rede pública de saúde, e analisar a interface das representações sociais da AIDS com o cotidiano dos indivíduos que vivem com o vírus da imunodeficiência humana (HIV), especialmente no processo de adesão ao tratamento. Realizaram-se entrevistas com 30 indivíduos soropositivos. Utilizou-se a técnica de análise de conteúdo manual. Da análise, emergiram seis categorias que retraduziram o cotidiano de soropositivos, permeados pelo estigma, preconceito, luta pela vida e a necessidade do uso contínuo de antirretrovirais. A AIDS foi assimilada a doenças crônicas como diabetes, evidenciando tendência para transformação da representação social da AIDS, substituindo a ideia de morte, por vida. Conclui-se que as pessoas que convivem com HIV estão mais otimistas devido aos tratamentos eficazes no controle da doença

Genetic topography and cortical cell loss in Huntington's disease link development and neurodegeneration

Cortical cell loss is a core feature of Huntington Disease (HD), beginning many years before clinical motor diagnosis, during the premanifest stage. However, it is unclear how genetic topography relates to cortical cell loss. Here, we explore the biological processes and cell types underlying this relationship and validate this using cell-specific post-mortem data. Eighty premanifest participants on average 15 years from disease onset and 71 controls were included. Using volumetric and diffusion MRI we extracted HD-specific whole brain maps where lower grey matter volume and higher grey matter mean diffusivity, relative to controls, were used as proxies of cortical cell loss. These maps were combined with gene expression data from the Allen Human Brain Atlas (AHBA) to investigate the biological processes relating genetic topography and cortical cell loss. Cortical cell loss was positively correlated with the expression of developmental genes (i.e. higher expression correlated with greater atrophy and increased diffusivity) and negatively correlated with the expression of synaptic and metabolic genes that have been implicated in neurodegeneration. These findings were consistent for diffusion MRI and volumetric HD-specific brain maps. As wild type Huntingtin is known to play a role in neurodevelopment, we explored the association between wild type Huntingtin (HTT) expression and developmental gene expression across the AHBA. Co-expression network analyses in 134 human brains free of neurodegenerative disorders was also performed. HTT expression was correlated with the expression of genes involved in neurodevelopment while co-expression network analyses also revealed that HTT expression was associated with developmental biological processes. Expression weighted cell-type enrichment (EWCE) analyses were used to explore which specific cell-types were associated with HD cortical cell loss and these associations were validated using cell specific single nucleus RNAseq (snRNAseq) data from post-mortem HD brains. The developmental transcriptomic profile of cortical cell loss in preHD was enriched in astrocytes and endothelial cells, while the neurodegenerative transcriptomic profile was enriched for neuronal and microglial cells. Astrocyte-specific genes differentially expressed in HD post-mortem brains relative to controls using snRNAseq were enriched in the developmental transcriptomic profile, while neuronal and microglial-specific genes were enriched in the neurodegenerative transcriptomic profile Our findings suggest that cortical cell loss in preHD may arise from dual pathological processes, emerging as a consequence of neurodevelopmental changes, at the beginning of life, followed by neurodegeneration in adulthood, targeting areas with reduced expression of synaptic and metabolic genes. These events result in age-related cell death across multiple brain cell types