Activation of pluripotency genes in human fibroblast cells by a novel mRNA based approach

Background: Several methods have been used to induce somatic cells to re-enter the pluripotent state. Viral transduction of reprogramming genes yields higher efficiency but involves random insertions of viral sequences into the human genome. Although induced pluripotent stem (iPS) cells can be obtained with the removable PiggyBac transposon system or an episomal system, both approaches still use DNA constructs so that resulting cell lines need to be thoroughly analyzed to confirm they are free of harmful genetic modification. Thus a method to change cell fate without using DNA will be very useful in regenerative medicine. Methodology/Principal Findings: In this study, we synthesized mRNAs encoding OCT4, SOX2, cMYC, KLF4 and SV40 large T (LT) and electroporated them into human fibroblast cells. Upon transfection, fibroblasts expressed these factors at levels comparable to, or higher than those in human embryonic stem (ES) cells. Ectopically expressed OCT4 localized to the cell nucleus within 4 hours after mRNA introduction. Transfecting fibroblasts with a mixture of mRNAs encoding all five factors significantly increased the expression of endogenous OCT4, NANOG, DNMT3 beta, REX1 and SALL4. When such transfected fibroblasts were also exposed to several small molecules (valproic acid, BIX01294 and 5'-aza-2'-deoxycytidine) and cultured in human embryonic stem cell (ES) medium they formed small aggregates positive for alkaline phosphatase activity and OCT4 protein within 30 days. Conclusion/Significance: Our results demonstrate that mRNA transfection can be a useful approach to precisely control the protein expression level and short-term expression of reprogramming factors is sufficient to activate pluripotency genes in differentiated cells

The production of immunity to Cl. chauvoei

1. Potent Cl. chauvoei vaccine (anaculture) may be made in horse-flesh infusion broth plus 1.0 per cent. peptone, 0.85 per cent. salt and 2-5 per cent. of serum and/or the haemolysed red cells of the horse, ox and sheep (meat particles one-third by volume). 2. Tryptic digests of horse muscle media produce satisfactory vaccines. 3. Boiled, dense suspensions of Cl. chauvoei injected subcutaneously into sheep produce a high degree of protection to culture injected intramuscularly. 4. The medium and method finally decided on as a routine measure was: ox-liver infusion broth (liver particles one-third by volume) plus 1.0 per cent. peptone, 0.85 per cent. salt and 1 per cent. glucose added after sterilization, pH 7.8, 36 hours’ incubation, and 0.5 per cent. formolization for three days at 37°C.The articles have been scanned in colour with a HP Scanjet 5590; 300dpi. Adobe Acrobat XI Pro was used to OCR the text and also for the merging and conversion to the final presentation PDF-format

History repeats? : the rise of the new middle classes in the developing world

1. Although the ability to detect chemical cues is widespread in many organisms, it is surprising how little is known about the role of chemical communication in avian life histories. Nowadays, growing evidence suggests that birds can use olfaction in several contexts. However, we still do not know the role of bird olfaction in one of the most important determinants of survival, predator detection. 2. Blue tits, Cyanistes caeruleus L., were exposed to chemical cues of: (i) mustelid (predator), (ii) quail (odorous control); or (iii) water (odourless control) inside the nest-box where they were provisioning 8-day-old nestlings. 3. We show that blue tits were able to detect the chemical cues and showed antipredatory behaviours to cope with the risk of predation. Birds delayed their entry to the nest-box, and they perched on the hole of the nest-box and refused to enter more times when they found predator scent than control scents inside the nest-box. In addition, birds decreased the time spent inside the predator-scented nest-box when feeding nestlings. 4. The discovery of the ability of birds to use chemical cues of predators to accurately assess predation may help to understand many aspects of bird life histories that have been neglected until now.Peer reviewe

Associations between sole ulcer, white line disease and digital dermatitis and the milk yield of 1824 dairy cows on 30 dairy cow farms in England and Wales from February 2003–November 2004

The milk yields of 1824 cows were used to investigate the effect of lesion-specific causes of lameness, based on farmer treatment and diagnosis of lame cows, on milk yield. A three level hierarchical model of repeated test day yields within cows within herds was used to investigate the impact of lesion-specific causes of lameness (sole ulcer, white line disease, digital dermatitis and other causes) on milk yield before and after treatment compared with unaffected cows. Cattle which developed sole ulcer (SU) and white line disease (WLD) were higher yielding cattle before they were diagnosed. Their milk production fell to below that of the mean of unaffected cows before diagnosis and remained low after diagnosis. In cattle which developed digital dermatitis (DD) there was no significant difference in milk yield before treatment and a slightly raised milk yield immediately after treatment. The estimated milk loss attributable to SU and WLD was approximately 570kg and 370kg respectively. These results highlight that specific types of lameness vary by herds and within herds they are associated with higher yielding cattle. Consequently lesion-specific lameness reduction programmes targeting the cow and farm specific causes of lameness might be more effective than generic recommendations. They also highlight the importance of milk loss when estimating the economic impact of SU and WLD on the farms profitability

Vertical transport and electroluminescence in InAs/GaSb/InAs structures: GaSb thickness and hydrostatic pressure studies

We have measured the current-voltage (I-V) of type II InAs/GaSb/InAs double heterojunctions (DHETs) with 'GaAs like' interface bonding and GaSb thickness between 0-1200 \AA. A negative differential resistance (NDR) is observed for all DHETs with GaSb thickness $>$ 60 \AA below which a dramatic change in the shape of the I-V and a marked hysteresis is observed. The temperature dependence of the I-V is found to be very strong below this critical GaSb thickness. The I-V characteristics of selected DHETs are also presented under hydrostatic pressures up to 11 kbar. Finally, a mid infra-red electroluminescence is observed at 1 bar with a threshold at the NDR valley bias. The band profile calculations presented in the analysis are markedly different to those given in the literature, and arise due to the positive charge that it is argued will build up in the GaSb layer under bias. We conclude that the dominant conduction mechanism in DHETs is most likely to arise out of an inelastic electron-heavy-hole interaction similar to that observed in single heterojunctions (SHETs) with 'GaAs like' interface bonding, and not out of resonant electron-light-hole tunnelling as proposed by Yu et al. A Zener tunnelling mechanism is shown to contribute to the background current beyond NDR.Comment: 8 pages 12 fig

Experiences of preparing children for a death of an important adult during the COVID-19 pandemic: a mixed methods study

Objective: The objectives of this study were to investigate how families prepared children for the death of a significant adult, and how health and social care professionals provided psychosocial support to families about a relative’s death during the COVID-19 pandemic. Design/setting: A mixed methods design; an observational survey with health and social care professionals and relatives bereaved during the COVID-19 pandemic in the UK, and in-depth interviews with bereaved relatives and professionals were conducted. Data were analysed thematically. Participants: A total of 623 participants completed the survey and interviews were conducted with 19 bereaved relatives and 16 professionals. Results: Many children were not prepared for a death of an important adult during the pandemic. Obstacles to preparing children included families’ lack of understanding about their relative’s declining health; parental belief that not telling children was protecting them from becoming upset; and parents’ uncertainty about how best to prepare their children for the death. Only 10.2% (n=11) of relatives reported professionals asked them about their deceased relative’s relationships with children. This contrasts with 68.5% (n=72) of professionals who reported that the healthcare team asked about patient’s relationships with children. Professionals did not provide families with psychosocial support to facilitate preparation, and resources were less available or inappropriate for families during the pandemic. Three themes were identified: (1) obstacles to telling children a significant adult is going to die, (2) professionals’ role in helping families to prepare children for the death of a significant adult during the pandemic, and (3) how families prepare children for the death of a significant adult. Conclusions: Professionals need to: provide clear and honest communication about a poor prognosis; start a conversation with families about the dying patient’s significant relationships with children; and reassure families that telling children someone close to them is dying is beneficial for their longer term psychological adjustment

Genomic DNA transposition induced by human PGBD5

Transposons are mobile genetic elements that are found in nearly all organisms, including humans. Mobilization of DNA transposons by transposase enzymes can cause genomic rearrangements, but our knowledge of human genes derived from transposases is limited. In this study, we find that the protein encoded by human PGBD5, the most evolutionarily conserved transposable element-derived gene in vertebrates, can induce stereotypical cut-and-paste DNA transposition in human cells. Genomic integration activity of PGBD5 requires distinct aspartic acid residues in its transposase domain, and specific DNA sequences containing inverted terminal repeats with similarity to piggyBac transposons. DNA transposition catalyzed by PGBD5 in human cells occurs genome-wide, with precise transposon excision and preference for insertion at TTAA sites. The apparent conservation of DNA transposition activity by PGBD5 suggests that genomic remodeling contributes to its biological function

Measuring CMB Polarization with BOOMERANG

BOOMERANG is a balloon-borne telescope designed for long duration (LDB) flights around Antarctica. The second LDB Flight of BOOMERANG took place in January 2003. The primary goal of this flight was to measure the polarization of the CMB. The receiver uses polarization sensitive bolometers at 145 GHz. Polarizing grids provide polarization sensitivity at 245 and 345 GHz. We describe the BOOMERANG telescope noting changes made for 2003 LDB flight, and discuss some of the issues involved in the measurement of polarization with bolometers. Lastly, we report on the 2003 flight and provide an estimate of the expected results.Comment: 12 pages, 8 figures, To be published in the proceedings of "The Cosmic Microwave Background and its Polarization", New Astronomy Reviews, (eds. S. Hanany and K.A. Olive). Fixed typos, and reformatted citation