In vitro differentiation of mouse granulocytes. Programmed Cell Death: Methods and Protocols

Granulocytes are central players of the immune system and, once activated, a tightly controlled balance between effector functions and cell removal by apoptosis guarantees maximal host benefit with least possible collateral damage to healthy tissue. Granulocytes are end-differentiated cells that cannot be maintained in culture for prolonged times. Isolating primary granulocytes is inefficient and challenging when working with mice, and especially so for the lowly abundant eosinophil and basophils subtypes. Here we describe an in vitro protocol to massively expand mouse derived myeloid progenitors and to differentiate them ‘on demand’ and in large numbers into mature neutrophils or basophils.

Methods for preparing and counting biochemical varves

Four different preparation and counting methods for biochemical varves were compared in order to assess counting errors and to standardize these techniques. The properties of two embedding methods, namely the shock-freeze, freeze-dry and the water-acetone-epoxy-exchange method, are discussed. Varve counts were carried out on fresh sediment and on sediment thin-sections, on the latter by manual and by automated counting using image-analysis software. Counting on fresh sediment and using image-analysis generally underestimated the number of varves, especially in sections with inconspicuous varves. A comparison between multiple varve counts carried out by a single analyst and different analysts showed no significant differences in the mean varve counts.

Complications of misoprostol and other abortion induction methods in the developing world: A systematic review

OBJECTIVE: To systematically review published literature to examine the complications associated with the use of misoprostol and compare these complications to those associated with other forms of abortion induction. ^ DATA SOURCES: Studies were identified through searches of medical literature databases including Medline (Ovid), PubMed (NLM), LILACS, sciELO, and AIM (AFRO), and review of references of relevant articles. ^ STUDY SELECTION AND METHODS: A descriptive systematic review that included studies reported in English and published before December 2012. Eligibility criteria included: misoprostol (with or without other methods) and any other method of abortion in a developing country, as well as quantitative data on the complication of each method. The following is information extracted from each study: author/year, country/city, study design/study sample, age range, setting of data collection, sample size, the method of abortion induction, the number of cases for each method, and the percentage of complications with each method. RESULTS: A total of 4 studies were identified (all in Latin America) describing post-abortion complications of misoprostol and other methods in countries where abortion is generally considered unsafe and/or illegal. The four studies reported on a range of complications including: bleeding, infection, incomplete abortion, intense pelvic pain, uterine perforation, headache, diarrhea, nausea, mechanical lesions, and systemic collapse. The most prevalent complications of misoprostol-induced abortion reported were: bleeding (7-82%), incomplete abortion (33-70%), and infection (0.8-67%). The prevalence of these complications reported from other abortion methods include: bleeding (16-25%), incomplete abortion (15-82%), and infection (13-50%). ^ CONCLUSION: The literature identified by this systematic review is inadequate for determining the complications of misoprostol used in unsafe settings. Abortion is considered an illicit behavior in these countries, therefore making it difficult to investigate the details needed to conduct a study on abortion complications. Given the differences between the reviewed studies as well as a variety of study limitations, it is not possible to draw firm conclusions about the rates of specific-abortion related complications.^

Discrete Multi-Criteria Methods for lands use and conservation planning on La Colacha in Arroyos Menores (Río Cuarto, Province of Córdoba, Argentina)

The study area is La Colacha sub-basins from Arroyos Menores basins, natural areas at West and South of Río Cuarto in Province of Córdoba of Argentina, fertile with loess soils and monsoon temperate climate, but with soil erosions including regressive gullies that degrade them progressively. Cultivated gently since some hundred sixty years, coordinated action planning became necessary to conserve lands while keeping good agro-production. The authors had improved data on soils and on hydrology for the study area, evaluated systems of soil uses and actions to be recommended and applied Decision Support Systems (DSS) tools for that, and were conducted to use discrete multi-criteria models (MCDM) for the more global views about soil conservation and hydraulic management actions and about main types of use of soils. For that they used weighted PROMETHEE, ELECTRE, and AHP methods with a system of criteria grouped as environmental, economic and social, and criteria from their data on effects of criteria. The alternatives resulting offer indication for planning depending somehow on sub basins and on selections of weights, but actions for conservation of soils and water management measures are recommended to conserve the basins conditions, actually sensibly degrading, mainly keeping actual uses of the lands.

Post-­‐occupancy monitoring of two flats in Madrid: development and assessment of a mixed methods methodology

Recent research has shown large differences between the expected and the actual energy consumption in buildings. The differences have been attributed partially, to the assumptions made during the design phase of buildings when simulation methods are employed. More accurate occupancy profiles on building operation could help to carry out more precise building performance calculations. This study focuses on the post-occupancy evaluation of two apartments, one renovated and one non renovated, in Madrid within the same building complex. The aim of this paper is to present an application of the mixed-methods methodology (Creswell, 2007) to assess thermal comfort and occupancy practices used in the case studies, and to discuss the shortcomings and opportunities associated with it. The mixed-methods methodology offers strategies for integrating qualitative and quantitative methods to investigate complex phenomena. This approach is expected to contribute to the growing knowledge of occupants’ behaviour and building performance by explaining the differences observed between energy consumption and thermal comfort in relation to people’s saving and comfort practices and the related experiences, preferences and values.

New Methods for Measuring Transient Interactions Between Proteins and Curved Membranes

Variations in the physical deformation of the plasma membrane play a significant role in the sorting and behavior of the proteins that occupy it. Determining the interplay between membrane curvature and protein behavior required the development and thorough characterization of a model plasma membrane with well defined and localized regions of curvature. This model system consists of a fluid lipid bilayer that is supported by a dye-loaded polystyrene nanoparticle patterned glass substrate. As the physical deformation of the supported lipid bilayer is essential to our understanding of the behavior of the protein occupying the bilayer, extensive characterization of the structure of the model plasma membrane was conducted. Neither the regions of curvature in the vicinity of the polystyrene nanoparticles or the interaction between a lipid bilayer and small patches of curved polystyrene are well understood, so the results of experiments to determine these properties are described. To do so, individual fluorescently labeled proteins and lipids are tracked on this model system and in live cells. New methods for analyzing the resulting tracks and ensemble data are presented and discussed. To validate the model system and analytical methods, fluorescence microscopy was used to image a peripheral membrane protein, cholera toxin subunit B (CTB). These results are compared to results obtained from membrane components that were not expected to show an preference for membrane curvature: an individual fluorescently-labeled lipid, lissamine rhodamine B DHPE, and another protein, streptavidin associated with biotin-labeled DHPE. The observed tendency for cholera toxin subunit B to avoid curved regions of curvature, as determined by new and established analytical methods, is presented and discussed.

Coastal Erosion--Causes, Erosion Control Methods, Economic and Environmental Impacts, and Best Management Practices

Coastal erosion is an important and constant issue facing coastal areas all over the world today. The rate of coastal development over the years has increased, in turn requiring that action be taken to protect structures from the threat of erosion. A review of the causes of coastal erosion and the methods implemented to control it was conducted in order to determine the best course of action in response to coastal erosion issues. The potential positive and negative economic and environmental impacts are key concerns in determining whether or not to restore an eroding beach and which erosion control method(s) to implement. Results focus on providing a comparison of these concerns as well as recommendations for addressing coastal erosion issues.

Thermogravimetry and Py–GC/MS techniques as fast qualitative methods for comparing the biochemical composition of Nannochloropsis oculata samples obtained under different culture conditions

Microalgae have many applications, such as biodiesel production or food supplement. Depending on the application, the optimization of certain fractions of the biochemical composition (proteins, carbohydrates and lipids) is required. Therefore, samples obtained in different culture conditions must be analyzed in order to compare the content of such fractions. Nevertheless, traditional methods necessitate lengthy analytical procedures with prolonged sample turn-around times. Results of the biochemical composition of Nannochloropsis oculata samples with different protein, carbohydrate and lipid contents obtained by conventional analytical methods have been compared to those obtained by thermogravimetry (TGA) and a Pyroprobe device connected to a gas chromatograph with mass spectrometer detector (Py–GC/MS), showing a clear correlation. These results suggest a potential applicability of these techniques as fast and easy methods to qualitatively compare the biochemical composition of microalgal samples.

Methods for the analysis of columbium and its alloys /

This report consists of the analytical procedures modified or developed at Pratt & Whitney Aircraft, CANEL, for the determination of alloying constituents and impurities in columbium and its alloys. Included are spectrophotometric methods for chromium, columbium, iron, molybdenum, tungsten, nickel, nitrogen and titanium; volumetric methods for chromium, vanadium and zirconium; emission and X-ray spectrographic methods for various alloying elements; a spectrographic method for zirconium and trace impurities and miscellaneous methods for aluminum, carbon, oxygen and hydrogen.

Your land : a handbook of practical methods for preventing and curing soil erosion.

"A compilation of practical tried-and-proved methods which have been developed by state agricultural colleges and extension workers, U.S. soil conservation experts, and county agricultural agents."--p. [2] of cover.

Advanced mean field methods:theory and practice

A major problem in modern probabilistic modeling is the huge computational complexity involved in typical calculations with multivariate probability distributions when the number of random variables is large. Because exact computations are infeasible in such cases and Monte Carlo sampling techniques may reach their limits, there is a need for methods that allow for efficient approximate computations. One of the simplest approximations is based on the mean field method, which has a long history in statistical physics. The method is widely used, particularly in the growing field of graphical models. Researchers from disciplines such as statistical physics, computer science, and mathematical statistics are studying ways to improve this and related methods and are exploring novel application areas. Leading approaches include the variational approach, which goes beyond factorizable distributions to achieve systematic improvements; the TAP (Thouless-Anderson-Palmer) approach, which incorporates correlations by including effective reaction terms in the mean field theory; and the more general methods of graphical models. Bringing together ideas and techniques from these diverse disciplines, this book covers the theoretical foundations of advanced mean field methods, explores the relation between the different approaches, examines the quality of the approximation obtained, and demonstrates their application to various areas of probabilistic modeling.

The comparative impact of privatisation and regulation on productivity growth in the English and Welsh water and sewerage industry, 1985-99

After the 10 regional water authorities of England and Wales were privatized in November 1989, the successor WASCs (water and sewerage companies) faced a new regulatory regime that was designed to promote productivity growth while simultaneously improving drinking water and environmental quality. As legally mandated quality improvements necessitated a costly capital investment programme, the industry's economic regulator – the Office of Water Services – implemented a RPI + K pricing system, designed to compensate the WASCs for their capital investment programme while also encouraging faster rates of productivity growth. This paper considers the relative effects of privatization and regulation on productivity growth in the industry using both non-parametric and parametric methods to provide a crosscheck on the robustness of the results. While there is evidence that labour productivity improved after privatization, there is no evidence that privatization led to a growth in TFP (total factor productivity). However, there is some evidence of a small increase in the rate of TFP growth in the aftermath of a substantial tightening of the regulatory regime that took place in 1995. These results, therefore, are consistent with evidence from other research that privatization, in the absence of effective competition and/or regulation, is not necessarily associated with improved economic performance.

Profitability of a sample of Portuguese bank branches and its decomposition into technical and allocative components

The efficiency literature, both using parametric and non-parametric methods, has been focusing mainly on cost efficiency analysis rather than on profit efficiency. In for-profit organisations, however, the measurement of profit efficiency and its decomposition into technical and allocative efficiency is particularly relevant. In this paper a newly developed method is used to measure profit efficiency and to identify the sources of any shortfall in profitability (technical and/or allocative inefficiency). The method is applied to a set of Portuguese bank branches first assuming long run and then a short run profit maximisation objective. In the long run most of the scope for profit improvement of bank branches is by becoming more allocatively efficient. In the short run most of profit gain can be realised through higher technical efficiency. © 2003 Elsevier B.V. All rights reserved.

Data analysis methods in optometry. Part 5: correlation

1. Pearson's correlation coefficient only tests whether the data fit a linear model. With large numbers of observations, quite small values of r become significant and the X variable may only account for a minute proportion of the variance in Y. Hence, the value of r squared should always be calculated and included in a discussion of the significance of r. 2. The use of r assumes that a bivariate normal distribution is present and this assumption should be examined prior to the study. If Pearson's r is not appropriate, then a non-parametric correlation coefficient such as Spearman's rs may be used. 3. A significant correlation should not be interpreted as indicating causation especially in observational studies in which there is a high probability that the two variables are correlated because of their mutual correlations with other variables. 4. In studies of measurement error, there are problems in using r as a test of reliability and the ‘intra-class correlation coefficient’ should be used as an alternative. A correlation test provides only limited information as to the relationship between two variables. Fitting a regression line to the data using the method known as ‘least square’ provides much more information and the methods of regression and their application in optometry will be discussed in the next article.

Recombinant protein production in yeast:methods and protocols

In the last few years, significant advances have been made in understanding how a yeast cell responds to the stress of producing a recombinant protein, and how this information can be used to engineer improved host strains. The molecular biology of the expression vector, through the choice of promoter, tag and codon optimization of the target gene, is also a key determinant of a high-yielding protein production experiment. Recombinant Protein Production in Yeast: Methods and Protocols examines the process of preparation of expression vectors, transformation to generate high-yielding clones, optimization of experimental conditions to maximize yields, scale-up to bioreactor formats and disruption of yeast cells to enable the isolation of the recombinant protein prior to purification. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.