Bacillus halodurans RecA-DNA binding and RecAmediated cleavage enhancing at alkaliphilic pH in vitro

In Escherichia coli, RecA protein catalyzes DNA pairing and strand exchange activities essential for genetic recombination. This is critical for normal cellular function under conditions that lead to alteredDNA metabolism and DNA damage. The RecA proteins of E. coli and Bacillus halodurans both can bind to DNA and catalyze the specific proteolytic cleavage of LexA and lambda repressor which inducesSOS response. At neutral pH self-cleavage of LexA depends exclusively on its binding to RecA filament, while at elevated pH (~10) it autodigests in the absence of RecA. We have shown in this work that the RecA-mediated cleavage and the binding of RecA to DNA promoted by B. halodurans are similar to those promoted by E. coli RecA, excepted that in the case of B. halodurans the rate of the cleavagereactions is increased at alkaline pH and that NaCl favors the binding of RecA to DNA. The results lead to two hypotheses for the pathway for RecA-mediated cleavage, in which we first suppose that theinternal pH of the bacteria is neutral. Thus LexA cannot undergo autodigestion, the genes involving in DNA repair and replication are not transcribed, regulating the growth of the cell. The second hypothesisis that the external environment and the internal pH of the bacteria are alkaline; here also the bacteria may have developed strategies to maintain LexA not inactivated. These observations suggest that theLexA autodigestion in B. halodurans at high pH may be regulated at the transcriptional level and that B. halodurans may be haloalkaliphile bacterium

A comparison of methods for determining ploidy in white sturgeon (Acipenser transmontanus)

Release of sturgeon with abnormal ploidy into the wild may result in reduced fitness due to lowered fertility in the F2 and subsequent generations. Further, there is evidence that ploidy affects reproductive development and caviar yield. Therefore, the ability to accurately characterize the ploidy of white sturgeon (Acipenser transmontanus)is essential for both commercial and conservation aquaculture. This study compares nuclear volume and whole erythrocyte long-axis lengths obtained using Coulter counter and blood smears, respectively, from captive white sturgeon from populations originating in California and Idaho. We examine which method provides the most accurate, time efficient and cost-effective characterization of ploidy in this species. Results from Coulter counter and blood smears were compared to results from flow cytometry, the gold standard for genome size analysis. Previous work suggests that blood smears can distinguish between 8N (diploid)and 12N (triploid)sturgeon, but further analysis is required to see if this method can also be used to identify 10N fish and to provide robust evidence of its utility in 8N and 12N fish across populations. In this study, we demonstrated that the Coulter counter had 100% agreement with flow cytometry in ploidy assignment, while blood smears vary in their accuracy based on population. Blood smears showed a high degree of overlap in erythrocyte long-axis length between 8N and 10N individuals as well as some overlap between 10N and 12N individuals in the California fish, and a high degree of overlap between 8N and 12N individuals in the Idaho fish. Although blood smears are time-intensive and vary in their ploidy assignment accuracy, they are a low-cost technique and as such may have some utility for caviar farms attempting to identify 12N individuals in a small number of broodstock. By comparing the accuracy, efficiency and cost of these three methods, sturgeon farmers and conservation hatcheries will be able to choose the best method for their needs in determining the ploidy of their fish. We determined that Coulter counter is equally accurate to flow cytometry and is also the most time efficient method for ploidy determination in white sturgeon

Demolition or Refurbishment of Social Housing? A review of the evidence

This report provides a review of technical models, evidence and case studies for decision making relating to the retention or demolition of social housing stock. Technical assessments of building suitability for refurbishment or demolition are often based on models of building performance. These include energy performance of the building compared to standards for new buildings, and assessment of environmental and energy impacts of the building over its lifetime from construction to demolition. Decisions can also be based on a series of performance and cost indicators. All modelling and indicator based approaches require assumptions about the building and the economic and policy context in which regeneration will take place, which need to be examined and justified in each case. Evaluation of the economic case for refurbishment is sensitive to the institutional factors such as the UK retrofit supply chain and market; tenure types and management capacity; access to finance and/or willingness to invest. Typical cost indicators are capital expenditure, operational and maintenance expenditure and capital investment appraisal. Estimating the costs and impacts of refurbishment or demolition is complex, uncertain and subjective – especially where nonmonetary costs and benefits have to be assigned a value. Finance mechanisms for refurbishment are less well established than for construction. The energy performance of a building is an increasingly important consideration in decisions to demolish or refurbish, and it has a big impact on the health of residents and the cost of their energy bills. Energy is used by residents as they live in a building throughout its lifetime. Energy is also used to manufacture building materials and construct the building in the first place and then in demolition, reusing, recycling and moving materials to dispose of them. Reducing carbon emissions associated with the built environment means reducing the emissions associated with the whole lifecycle of buildings. However, refurbishment and retrofitting of buildings, including insulation, replacing windows and boilers, heating networks, and installing renewable energy, can improve the performance of existing buildings to near-new standards. Decarbonising the UK electricity grid will also reduce the climate change impacts of energy used in buildings, and will increase the relative importance of embodied carbon and energy in the lifecycle impacts of a building. Case studies demonstrate that even older, high rise or poorly insulated structures, known as hard to treat buildings, can be retrofitted to achieve high energy efficiency standards. In these cases, the costs of retrofitting compared to demolition and new construction can also be lower, particularly when construction work has been organised so that residents have been able to stay in their homes avoiding the costs and disruption of temporarily housing people elsewhere. The carbon emissions associated with building use depend on the source of energy used. Increased low carbon sources of energy to produce electricity on the grid in the future may reduce the environmental impacts of energy used in homes. Research has shown that there are often differences between the predicted and actual performance of buildings (performance gaps) and that people sometimes adapt their behaviour in ways that increase consumption after an energy efficiency project (rebound effects). Performance gaps and rebound effects are often not taken into account when assessing benefits to residents like a reduction in bills or improvements in thermal comfort. If future savings have been over-estimated, it is residents (rather than the professionals estimating the savings) who are doubly and disproportionately penalised, firstly, because what has been promised is not delivered and, secondly, because they pay the energy bills. Relatively simple water efficiency retrofitting can achieve savings of 17.5 litres per person per day, compared with the London average of 160 litres of water used per person per day. Sustainable drainage methods can also be cost effectively retrofitted into existing buildings and estates, delivering a wide range of benefits including reduced overheating of buildings. The construction and demolition sector contributes 35% of all waste in the UK every year. Much of this is due to demolition waste. The UK construction sector currently recycles 73% of its waste, but still contributes more than 4 million tonnes of waste to landfill each year. Recycling demolition waste reduces the environmental impacts of demolition, but refurbishment avoids waste to landfill and many of the environmental impacts of new construction. Improving the quality of social housing stock is essential to reduce health inequalities in the UK. Housing has significant impacts on mental and physical health and wellbeing, and should be a key factor in regeneration decision making. Refurbishment can deliver improvements in housing quality at a faster rate than demolition and rebuilding of social housing, but health issues such as ventilation and indoor air quality can be complex issues to address in refurbishment. Refurbishment of buildings presents opportunities for the creation of jobs requiring a new set of skills that will be in demand if the UK is to meet its carbon emission reduction targets. Operation of renewable energy systems also provides opportunities for community development through refurbishment of buildings and estates. It is clear that the ability for communities to engage in refurbishment and demolition decisions would be enhanced by a consistent and transparent approach to the reporting of lifecycle costs, energy and carbon, water and waste and monitoring the wellbeing of those affected by refurbishment and demolition. The literature reviewed here is emerging from different fields – engineers, energy modellers, planners and public health specialists – and shows some useful results but is often hard to disaggregate in a way that shows how the effects of refurbishment and demolition play out for different groups of people. However, many aspects of refurbishment and demolition are complex and interact with each other: what is needed is a more balanced inter-disciplinary view of what housing interventions mean for people, and who the winners and losers are in the short and longer term

Increasing confidence and changing behaviors in primary care providers engaged in genetic counselling.

BackgroundScreening and counseling for genetic conditions is an increasingly important part of primary care practice, particularly given the paucity of genetic counselors in the United States. However, primary care physicians (PCPs) often have an inadequate understanding of evidence-based screening; communication approaches that encourage shared decision-making; ethical, legal, and social implication (ELSI) issues related to screening for genetic mutations; and the basics of clinical genetics. This study explored whether an interactive, web-based genetics curriculum directed at PCPs in non-academic primary care settings was superior at changing practice knowledge, attitudes, and behaviors when compared to a traditional educational approach, particularly when discussing common genetic conditions.MethodsOne hundred twenty one PCPs in California and Pennsylvania physician practices were randomized to either an Intervention Group (IG) or Control Group (CG). IG physicians completed a 6 h interactive web-based curriculum covering communication skills, basics of genetic testing, risk assessment, ELSI issues and practice behaviors. CG physicians were provided with a traditional approach to Continuing Medical Education (CME) (clinical review articles) offering equivalent information.ResultsPCPs in the Intervention Group showed greater increases in knowledge compared to the Control Group. Intervention PCPs were also more satisfied with the educational materials, and more confident in their genetics knowledge and skills compared to those receiving traditional CME materials. Intervention PCPs felt that the web-based curriculum covered medical management, genetics, and ELSI issues significantly better than did the Control Group, and in comparison with traditional curricula. The Intervention Group felt the online tools offered several advantages, and engaged in better shared decision making with standardized patients, however, there was no difference in behavior change between groups with regard to increases in ELSI discussions between PCPs and patients.ConclusionWhile our intervention was deemed more enjoyable, demonstrated significant factual learning and retention, and increased shared decision making practices, there were few differences in behavior changes around ELSI discussions. Unfortunately, barriers to implementing behavior change in clinical genetics is not unique to our intervention. Perhaps the missing element is that busy physicians need systems-level support to engage in meaningful discussions around genetics issues. The next step in promoting active engagement between doctors and patients may be to put into place the tools needed for PCPs to easily access the materials they need at the point-of-care to engage in joint discussions around clinical genetics

SNAI1 and SNAI2 Are Asymmetrically Expressed at the 2-Cell Stage and Become Segregated to the TE in the Mouse Blastocyst

SNAI1 and SNAI2 are transcription factors that initiate Epithelial-to-Mesenchymal cell transitions throughout development and in cancer metastasis. Here we show novel expression of SNAI1 and SNAI2 throughout mouse preimplantation development revealing asymmetrical localization of both SNAI1 and SNAI2 in individual blastomeres beginning at the 2-cell stage through to the 8-cell stage where SNAI1 and SNAI2 are then only detected in outer cells and not inner cells of the blastocyst. This study implicates SNAI1 and SNAI2 in the lineage segregation of the trophectoderm and inner cell mass, and provides new insight into these oncogenes

Ultrasonic Nondestructive Evaluation Using Laser Transducers

A program is described which employs lasers for ultrasonic NDE. A high-power laser is used to generate a brief sound pulse in the test specimen. A second low-power laser then measures the response of the specimen to that sound pulse. The response of the specimen is measured by a “Laser Vibrometer.” This is a novel type of heterodyne interferometer which focuses a Helium-Neon laser beam onto the surface of the specimen and measures its displacement. Displacements as small as 2×10-12 meters on a 0.15 sec averaging time can be detected and also displacements of 1.5×l0-9 meters on a 10-MHz bandwidth. The Laser Vibrometer has a well defined frequency response and does not introduce distortion. The sound generating laser is either a pulsed carbon dioxide TEA laser or a YAG laser. The peak power exceeds 10 M watt. Two mechanisms for generating the sound are discussed. The thermoelastic mechanism relies on the thermal expansion of the surface, causing it to move. The reaction to this causes a pressure pulse in the specimen. Another mechanism allows a small amount of the surface to be ablated and the reaction to this causes a substantial pressure pulse in the specimen. Both laser beams can be scanned over the surface of the specimen by a microprocessor controlled mirror. The microprocessor generates a raster scan of arbitrary size, number of lines, step size and speed. Eventually this technique will allow the inspection of complex specimens without direct contact. This will eliminate the tedium and contact reliability problems associated with conventional piezo-ceramic NDE

Gaps in detailed knowledge of human papillomavirus (HPV) and the HPV vaccine among medical students in Scotland

<p>Background: A vaccination programme targeted against human papillomavirus (HPV) types16 and 18 was introduced in the UK in 2008, with the aim of decreasing incidence of cervical disease. Vaccine roll out to 12–13 year old girls with a catch-up programme for girls aged up to 17 years and 364 days was accompanied by a very comprehensive public health information (PHI) campaign which described the role of HPV in the development of cervical cancer.</p> <p>Methods: A brief questionnaire, designed to assess acquisition of knowledge of HPV infection and its association to cervical cancer, was administered to two different cohorts of male and female 1st year medical students (school leavers: 83% in age range 17–20) at a UK university. The study was timed so that the first survey in 2008 immediately followed a summer's intensive PHI campaign and very shortly after vaccine roll-out (150 students). The second survey was exactly one year later over which time there was a sustained PHI campaign (213 students).</p> <p>Results: We addressed three research questions: knowledge about three specific details of HPV infection that could be acquired from PHI, whether length of the PHI campaign and/or vaccination of females had any bearing on HPV knowledge, and knowledge differences between men and women regarding HPV. No female student in the 2008 cohort had completed the three-dose vaccine schedule compared to 58.4% of female students in 2009. Overall, participants’ knowledge regarding the sexually transmitted nature of HPV and its association with cervical cancer was high in both year groups. However, in both years, less than 50% of students correctly identified that HPV causes over 90% of cases of cervical cancer. Males gave fewer correct answers for these two details in 2009. In 2008 only around 50% of students recognised that the current vaccine protects against a limited subset of cervical cancer-causing HPV sub-types, although there was a significant increase in correct response among female students in the 2009 cohort compared to the 2008 cohort.</p> <p>onclusions: This study highlights a lack of understanding regarding the extent of protection against cervical cancer conferred by the HPV vaccine, even among an educated population in the UK who could have a vested interest in acquiring such knowledge. The intensive PHI campaign accompanying the first year of HPV vaccination seemed to have little effect on knowledge over time. This is one of the first studies to assess detailed knowledge of HPV in both males and females. There is scope for continued improvements to PHI regarding the link between HPV infection and cervical cancer.</p&gt

Massive rearrangements of cellular MicroRNA signatures are key drivers of hepatocyte dedifferentiation

Hepatocytes are dynamic cells that, upon injury, can alternate between nondividing differentiated and dedifferentiated proliferating states in vivo . However, in two‐dimensional cultures, primary human hepatocytes (PHHs) rapidly dedifferentiate, resulting in loss of hepatic functions that significantly limits their usefulness as an in vitro model of liver biology, liver diseases, as well as drug metabolism and toxicity. Thus, understanding the underlying mechanisms and stalling of the dedifferentiation process would be highly beneficial to establish more‐accurate and relevant long‐term in vitro hepatocyte models. Here, we present comprehensive analyses of whole proteome and transcriptome dynamics during the initiation of dedifferentiation during the first 24 hours of culture. We report that early major rearrangements of the noncoding transcriptome, hallmarked by increased expression of small nucleolar RNAs, long noncoding RNAs, microRNAs (miRNAs), and ribosomal genes, precede most changes in coding genes during dedifferentiation of PHHs, and we speculated that these modulations could drive the hepatic dedifferentiation process. To functionally test this hypothesis, we globally inhibited the miRNA machinery using two established chemically distinct compounds, acriflavine and poly‐l ‐lysine. These inhibition experiments resulted in a significantly impaired miRNA response and, most important, in a pronounced reduction in the down‐regulation of hepatic genes with importance for liver function. Thus, we provide strong evidence for the importance of noncoding RNAs, in particular, miRNAs, in hepatic dedifferentiation, which can aid the development of more‐efficient differentiation protocols for stem‐cell‐derived hepatocytes and broaden our understanding of the dynamic properties of hepatocytes with respect to liver regeneration. Conclusion: miRNAs are important drivers of hepatic dedifferentiation, and our results provide valuable information regarding the mechanisms behind liver regeneration and possibilities to inhibit dedifferentiation in vitro

Air–sea exchange of acetone, acetaldehyde, DMS and isoprene at a UK coastal site

Volatile organic compounds (VOCs) are ubiquitous in the atmosphere and are important for atmospheric chemistry. Large uncertainties remain in the role of the ocean in the atmospheric VOC budget because of poorly constrained marine sources and sinks. There are very few direct measurements of air–sea VOC fluxes near the coast, where natural marine emissions could influence coastal air quality (i.e. ozone, aerosols) and terrestrial gaseous emissions could be taken up by the coastal seas. To address this, we present air–sea flux measurements of acetone, acetaldehyde and dimethylsulfide (DMS) at the coastal Penlee Point Atmospheric Observatory (PPAO) in the south-west UK during the spring (April–May 2018). Fluxes of these gases were measured simultaneously by eddy covariance (EC) using a proton-transfer-reaction quadrupole mass spectrometer. Comparisons are made between two wind sectors representative of different air–water exchange regimes: the open-water sector facing the North Atlantic Ocean and the terrestrially influenced Plymouth Sound fed by two estuaries. Mean EC (± 1 standard error) fluxes of acetone, acetaldehyde and DMS from the open-water wind sector were −8.0 ± 0.8, −1.6 ± 1.4 and 4.7 ± 0.6 µmol m−2 d−1 respectively (“−” sign indicates net air-to-sea deposition). These measurements are generally comparable (same order of magnitude) to previous measurements in the eastern North Atlantic Ocean at the same latitude. In comparison, the Plymouth Sound wind sector showed respective fluxes of −12.9 ± 1.4, −4.5 ± 1.7 and 1.8 ± 0.8 µmol m−2 d −1. The greater deposition fluxes of acetone and acetaldehyde within the Plymouth Sound were likely to a large degree driven by higher atmospheric concentrations from the terrestrial wind sector. The reduced DMS emission from the Plymouth Sound was caused by a combination of lower wind speed and likely lower dissolved concentrations as a result of the estuarine influence (i.e. dilution). In addition, we measured the near-surface seawater concentrations of acetone, acetaldehyde, DMS and isoprene from a marine station 6 km offshore. Comparisons are made between EC fluxes from the open-water and bulk air–sea VOC fluxes calculated using air and water concentrations with a two-layer (TL) model of gas transfer. The calculated TL fluxes agree with the EC measurements with respect to the directions and magnitudes of fluxes, implying that any recently proposed surface emissions of acetone and acetaldehyde would be within the propagated uncertainty of 2.6 µmol m−2 d −1. The computed transfer velocities of DMS, acetone and acetaldehyde from the EC fluxes and air and water concentrations are largely consistent with previous transfer velocity estimates from the open ocean. This suggests that wind, rather than bottom-driven turbulence and current velocity, is the main driver for gas exchange within the open-water sector at PPAO (depth of ∼ 20 m)

Reliability of Rapid Diagnostic Tests in Diagnosing Pregnancy-Associated Malaria in North-Eastern Tanzania.

Accurate diagnosis and prompt treatment of pregnancy-associated malaria (PAM) are key aspects in averting adverse pregnancy outcomes. Microscopy is the gold standard in malaria diagnosis, but it has limited detection and availability. When used appropriately, rapid diagnostic tests (RDTs) could be an ideal diagnostic complement to microscopy, due to their ease of use and adequate sensitivity in detecting even sub-microscopic infections. Polymerase chain reaction (PCR) is even more sensitive, but it is mainly used for research purposes. The accuracy and reliability of RDTs in diagnosing PAM was evaluated using microscopy and PCR. A cohort of pregnant women in north-eastern Tanzania was followed throughout pregnancy for detection of plasmodial infection using venous and placental blood samples evaluated by histidine rich protein 2 (HRP-2) and parasite lactate dehydrogenase (pLDH) based RDTs (Parascreen™) or HRP-2 only (Paracheck Pf® and ParaHIT®f), microscopy and nested Plasmodium species diagnostic PCR. From a cohort of 924 pregnant women who completed the follow up, complete RDT and microscopy data was available for 5,555 blood samples and of these 442 samples were analysed by PCR. Of the 5,555 blood samples, 49 ((proportion and 95% confidence interval) 0.9% [0.7 -1.1]) samples were positive by microscopy and 91 (1.6% [1.3-2.0]) by RDT. Forty-six (50.5% [40.5 - 60.6]) and 45 (49.5% [39.4 - 59.5]) of the RDT positive samples were positive and negative by microscopy, respectively, whereas nineteen (42.2% [29.0 - 56.7]) of the microscopy negative, but RDT positive, samples were positive by PCR. Three (0.05% [0.02 - 0.2]) samples were positive by microscopy but negative by RDT. 351 of the 5,461 samples negative by both RDT and microscopy were tested by PCR and found negative. There was no statistically significant difference between the performances of the different RDTs. Microscopy underestimated the real burden of malaria during pregnancy and RDTs performed better than microscopy in diagnosing PAM. In areas where intermittent preventive treatment during pregnancy may be abandoned due to low and decreasing malaria risk and instead replaced with active case management, screening with RDT is likely to identify most infections in pregnant women and out-performs microscopy as a diagnostic tool