bluuebunny/crossref_metadata_2025 huggingface dataset:
Reviewer Acknowledgements for English Language Teaching, Vol. 9, No. 1
Authors: Yu Gavin
Published: January 2016

Reviewer acknowledgements for English Language Teaching, Vol. 9, No. 1, 2016
Building a Strategy to Overcome the Psychological Barriers to Climate Change Management in Rural Communities of Fako Division, Cameroon
Authors: Kimengsi Jude Ndzifon , Gur Amawa Sani , Lydia Fondufe Sakah
Published: January 2016

This study seeks to build a strategy to overcome the psychological barriers to climate change management for rural communities in Fako Division. We employ a five point likert scale in which 100 inhabitants (adults) were surveyed purposefully surveyed in four rural communities (Malende, Bakingili, Bokwai and Miselele) of Fako Division (25 for each community) to identify the observed barriers. Based on the mean values derived from the 5 point likert scale, the study revealed that ignorance (mean=3.27) was the highest psychological barrier while denial stood as the least (mean=2.25). We then, as a recommendation, proposed a strategy for overcoming these psychological barriers which suggests that the government, the councils, NGOs, traditional authorities and the local population should collectively work together to identify people’s socio-economic needs and improve climate change management by empowering the population through workshop sensitisation, seminars and the use of the local media to reduce ignorance. Also, we suggest that they should motivate and create a number of incentives which would assist in reducing these observed barriers so as to ensure that developmental activities should respect stricto senso, issues of climate change management.
“The Commitment Problem”: Spending to Zero to Maximize the Efficiency of the Collections Budget
Authors: Cleary Robert M.
Published: January 2016

The difficulty for a library to spend their collections budget efficiently is a timeless problem. The diversity of a typical budget, with its mix of one-time and continuing funds, for an array of resources that have both regular and sometimes irregular frequencies, provides great challenges. Approval plans, usually expending one-time funds, generate expenditures that contain high variability on a weekly basis. Standing orders for serials fall into the same category. With some effort, it is possible to expend all continuing funds. But it is the commitments that do not result in expenditures, with funds remaining in cash balances that can determine what university administrators call “efficient results.” Acquisitions personnel must take an aggressive approach to commitments with the goal of turning as many possible into expenditures. New expenditures will compensate for the orders that remain committed. Based on the assumption that efficient spending focuses on a library budget’s final cash balance, this article presents a method to consistently achieve a zero or negative cash balance.
Armenia, the Armenians: between memory and future
Authors: Macioti Maria Immacolata
Published: January 2016

The article recalls how in 2015, the year of the centenary of the Armenian Genocide, important publications emerged, shedding a light on the incident that will be difficult to ignore. In particular, it was the archives of the Congregation for the Oriental Churches, the Vatican Secret Archives and the Historical Archive of the Secretariat of State which offered up precious material for the knowledge of what was happening to Armenians in Turkey to Georges-Henri Ruyssen SJ. To the latter is owed a valuable and significant work of reconstruction undertaken over many years, and consisting of eight volumes. These contain letters written in Turkey by those in contact with the people, who with their own eyes saw what was happening, and who followed what was occurring with dismay. Their relationship with the territory and their direct experience are fundamental. The correspondences begin in the years 1894-1896, of which volume IV, ranging from 1908 to 1925, is drawn precisely from the Secret Vatican Archives(ASV) as well as the Historic Archives of the Secretariat of State, the Section for Relations with Territories (SS.RR.SS). The author also points out a small valuable contribution by Jewish intellectuals of the time, translated into Italian with the title Pro Armenia. She underlines how, immediately, some Jews tried to intervene, to stop the genocide, and to assist Armenians in various ways. Today, it is difficult to ignore what happened, to ignore and to reduce what historically is the first genocide of the twentieth century in Europe. An event which has been minimized and denied: whose effects are still being felt today, and which has left a trail at individual, family and social levels. Armenian terrorism, which has resulted in attacks and deaths in Turkey and elsewhere, derives from these negations. The same is the case for a strong nationalism still prevalent today in the Republic of Armenia, which is constrained by circumstances to adopt a defensive political stance and – out of necessity – to maintain good relations with Russia. We have to wish that, after those resounding acknowledgement of the genocide, Armenia will be able to look forward to a more serene future.
The Specialist Court for Kosovo: continuity or departure from the hybrid courts model?
Authors: Selimi Shkёlzen
Published: January 2016

The issue of whether to establish Specialist Chambers within the Kosovo justice system for alleged war crimes committed in Kosovo has been, arguably, one of the most heated debates not only from a political and social point of view, but also from a legal one. While the required amendments in the Constitution and several laws of Kosovo necessary to establish the Specialist Chambers in furtherance of the agreement dated 14 April 2014 between the Republic of Kosovo and the European Union on the Mission of the European Union Rule of Law Mission in Kosovo (“EULEX”) will certainly create heated debates in the political level, one may argue that the legal issues that are expected to be encountered when the Specialist Chambers will be operational, may be even more pressing. This is in consideration of the peculiar nature of the Specialist Chambers, which are meant to have their basis within the laws of Kosovo, but at the same time, be independent from them and from control of Kosovo authorities. The purpose of this article is to delineate the possible legal issues that might confront the Specialist Chambers of Kosovo. Its main argument is that, while the Specialist Chambers seem to follow the experience of other hybrid internationalised courts, it still differs from them in some aspects. The challenges that the new Specialist Chambers may need to tackle deal with its jurisdiction and position within the Kosovo Judicial system, and its legitimacy and legal basis.
Inflation, exchange rate and efficacy of monetary policy in Nigeria: The empirical evidence
Authors: Ogbonna BigBen Chukwuma
Published: January 2016

This paper analyses the post SAP persistence of inflation in Nigeria for the period, 1960-2008 with exchange rate, money supply and trade balance as preferred influential variables. To investigate the effect of policy switch from the period of direct instrument to the period of deregulation occasioned by SAP of 1986, we proceed to bifurcate the sample period into two, comprising the period of direct instruments of monetary policy, 1960 – 1985 and the period of indirect instruments of monetary policy, 1986 – 2008. Estimates from a vector auto regression model (VAR) of key macroeconomic variables demonstrate the weak link between money supply and inflation in the both time horizons, which suggests that the hypothesis that money supply is not an effective policy instrument for management of inflationary developments cannot be rejected for Nigeria. The results further suggest that in both time horizons, exchange rate has been identified as a singular most promising macroeconomic fundamental for both internal and external sectors adjustments. However, the deregulation of the domestic economy as occasioned by SAP has significantly diluted the efficacy of exchange rate as a monetary policy instrument for the management of Nigeria’s aggregate money stock and trade balance developments. These notwithstanding, the Central Bank of Nigeria can continue to play a stabilizing role in the economy through the continuation of prudent monetary policies and frequent interventions in exchange rate management to smooth out shocks.
Transitology - A philosophical reflection on the post-communist society
Authors: Leka Agim
Published: January 2016

The collapse of totalitarianisms, the integration and transition global are objective phenomena, which changed many political and social doctrines, which relate to democracy and civilization. A review of relations between civilization and polity is most necessary. At the same time this process was companied with the crisis philosophical thought, as with as the need for a deep reflection in this area. Transition is the moving integration of society from ideology to the reality culture (the values system) and the integration of three truths: philosophical, scientific and religious. And the most effective vehicle is the cultural intercommunication. In global era the integration is a cultural intercommunications and the philosophy itself is an essential cultural. The dialectic of transition and integration in global time. The cultural shock and the vulgar hedonism. Paradox between interest and principles. The paradox of modern Transitology is it between time and space, or, between past time and future time, has made that many today’s society to be gripped by a severe conflict between the two sides: the old and new; the political right and the political left. The moral paradox or the paradox of “two twin brothers”.
Judicial control of administrative activity and advantages of reorganization of the juridical system in the Republic of Kosovo
Authors: Muçaj Florent , Gruda Përparim
Published: January 2016

Scientific research paper entitled “Judicial control of administrative activity and advantages of reorganization of the judicial system in the Republic of Kosovo” is treated with standard writing, including the introductory part and corresponding chapters. In the introductory part of the paper is emphasized the importance of the topic which is treated, with particular emphasis on the importance of judicial control in the Republic of Kosovo. This paper aims to achieve three main goals: Firstly, highlighting the importance and ways of functioning of judicial control in general, secondly, explication the method of treatment and the importance of the application of judicial control in the Republic of Kosovo and thirdly, highlighting the challenges of judicial control in the Republic of Kosovo, especially after the reorganization of the judicial system. The paper is structured as follows: introduction, general views on judicial control, including the importance of judicial control. Within the structure, important theses constitute those theses dedicated to administrative justice in the Republic of Kosovo, the legislative framework and judicial system reform in the Republic of Kosovo, including its impact on the functioning of the administrative judiciary. In the last part of the paper there are clear and consistent conclusions and significant recommendations relating to general views about judicial control, with particular emphasis on their practical implementation in the Republic of Kosovo and the way of adjustment with the reforms in the field of administrative justice which are at the beginnings of the implementation.
Energetic diplomacy and its role on creation of a new multipolar world
Authors: Gelev Igor , Racaj Muhamet
Published: January 2016

International correlations through the last years have been remarked as dramatic in character, and very often followed through with unpredictable events, such as the Arab spring, chronic and confessional wars raging through the Middle east, than the unprincipled informal collation of the great forces merging with variety and different in character formal and less formal state-of-actors, than worth mentioning is the recent war prelude in Ukraine which on top has all been spiced and made far more constrictive indeed by the arrival of the global economic crisis in energy resources as well as naturally found ones, that has well paid so far contributing led to the overflowing of the crisis itself from the world’s center towards its margins. Since the end of the cold war, the most powerful and the most economically ascendant countries have started to promote a very new breed type of concept which deals with export and imposing of its own ideology. Diplomacy has well been turned into an instrument for realization of the external affair politics of the great powers and this diplomacy has been thus transformed from traditional in character to modern in origin, or better - public diplomacy. But this breed of public diplomacy had its own limits in terms of its range influence which was shortcut in the days of lack of energy resources as well as naturally found pack resources in the sense of energy excavation, and thus we witnessed the arrival of the newly formed type of diplomacy in place of the public diplomacy, named energetic diplomacy. Today energetic diplomacy is a representative of modern diplomacy or better a unique type of “stick and carrot” modulated for the weak and well developed rich states which are dependent on energy resources naturally found or not. During the era of the predomination of traditional politics over power in international correlations, it was then the most important thing who would be the one to deal with the most military power arsenal and later economic power as well that determined supreme leading force. Many of us would ask does this mean the reincarnation of the so called almost forgotten Cold War only now coming in such a shape that seems to be far more terrifying in character with a great deal of chance to impose the brink of a new world war and another humanitarian wash-out to humanity.
European Council’s member states’ jurisdiction regarding the execution of court decisions and it’s issues
Authors: Gazidede Alkelina
Published: January 2016

European Court of Human Rights states that the execution of final decisions is a compulsory provision for a fair hearing and a successful conlusion of a trial. The right to a court protected by Article 6 would be illusory if a Contracting State’s domestic legal system allowed a final, binding judicial decision to remain inoperative to the detriment of one party. Execution of a judgment given by any court, is considered to be an integral part of the “trial” for the purposes of Article 6. Based on the Article 1 of European Convention for Human Rights member states of European Council are obligated to implement the the decisions of ECHR in their domestic legislation in the context of building an efficient legal system. This paper aimes to analyze the legal system of member states regarding the reinforcment of decisions looking forword to identify issues, commonalities and diferences among states. What is the procedure followed in the process of execution? What do we understand with “Reasonable timeframe” and which are the legal requirements in which are based “Concrete deadlines” within a judicial decision should be executed? What is the significance of the enforcment agents in this process?
From Welfare to Well-being: new approaches for a good life
Authors: Selvaggio Maria Antonietta
Published: January 2016

In this paper we reflect on some analysis conducted by researchers as Laura Balbo, Mary Catherine Bateson and Martha Nussbaum who, although having not had direct relations among them or specific partnerships, have greatly contributed to elaborate concepts and categories regarding our new vision of the well-being. This concept, so dear to the economist Amartya Sen, draws nourishment from thoughts and lived belonging to women’s life experience, particularly as concerning their capacity to organize their daily and family life, taking care of others, shouldering the vulnerabilities and building their biographical paths with creativity and adaptation. So we can understand what is a good life through the realization of our abilities, that is of each person, and not using a utility calculation. This involves a new approach to life quality and human development, that is exactly the capabilities approach described by Sen, rehashed by Nussbaum but, for some aspects, it was already present into the studies which characterize the gender prospective.
An analysis of the sources of competition disipline in the European Union and in Albania
Authors: Katro Eriona , Zhezha Vaeld
Published: January 2016

Competition discipline in the European Union is a substantial component of the EU’s institutional framework. The Lisbon Treaty made the “internal market” a shared competence, while necessary competition rules for the internal market are an exclusive competence of the Union. This paper aims to analyse the sources of the competition discipline in the EU and in Albania focusing more on constitutional sources. From this analyses will emerge that sources of competition discipline are numerous and each of them plays an important role in the development of the competition policy. But arises the question if these sources are all equally important or not? This paper will focus also n the importance of each source in both levels: European Union and Albanian discipline of competition.
Analysis of risks and investments’ opportunities in water sector
Authors: Zeneli Fjona
Published: January 2016

While water demand is projected to grow by 41% by 2030, considering also the ultimate reserves of drinking water, it is believed that this element will attract the majority of investments in the coming decades. Opportunities in drinking water sector are numerous, because its process of providing and delivery of drinking water includes many aspects: management of infrastructure, design of technological solutions, conservation and water’s quality. These opportunities result from the difference between water supply and water demand; an increasingly difference that requires capital investments in production and water treatment technologies. Investments need to be combined with the knowledge on the legislation, regulatory framework and technological developments. This article may serve to clarify type of investments in drinking water sector, known by literature, to identify opportunities of investment in this sector, indicating the theoretical framework of beta and alpha risk ratio coefficient calculation and to suggest how these types of investments can be allocated to the investment portfolios.
The council of the employees in the Albanian Commercial Law
Authors: Biba Diana
Published: January 2016

Law No. 9901/2008 on “Entrepreneurs and Commercial Companies”, marked a milestone in the reform of the commercial law in Albanian. Among other novelties, the Law introduced a new approach in regard to the employees and their participation in co determination. Actually, the involvement of the employees were not that unfamiliar in the former Albanian Commercial Legislation thought the social responsibility was. The Law brought in a new approach which was widely inspired from the EU Law, by establishing the Council of the Employees for any commercial company having more than 50 employees. It is true that unlike the trade unions, it is the company that bears the costs of the establishment and functioning of these councils, but besides the costs, it would mean to grant importance to the employees, as stakeholders of the corporate, by being part of the decision making process with regard to the use of special funds or actives of the company or to the allocation of the divided that the General Assembly resolves to allocate to the employees. This article will explore the legal provisions of the Law in regard to the Council of the Employees, its establishment, functioning and entitlement and how these provisions are enforced in practice from the companies in Albania.
Theoretical and practical implementation of the right for an equal enjoyment of health care service
Authors: Mano Laureta
Published: January 2016

This study is divided in three parts which intend to meet the following targets: Primarily to provide theoretical approaches toward the concept of equality in the healthcare. In the theoretical treatment of the concept of equality in health care the authors are cautious while making the difference between equality and justice; between equality in levels of health or health care and equity in financing etc. Secondly target it to assess the concept implemented in our legislation and make an overall assessment of whether this legal framework is comprehensive and convenient for the realization of the right to health care enshrined in the Constitution. The Albanian legislation stipulates that health care in the Republic of Albania is guided by very important principles. These principles embody the Constitution predictions from the formal point of view, primarily when declaring the right to health care as a key right of the individual, as well as when guaranteeing the equal rights based on non-discrimination. Thirdly to determine some of the specific areas of the legislation, which have to be improved and completed. After the analysis make in this paper, we found the need for some theoretical and practical improvements, which are reflected in the conclusions.
Length of proceedings as standard of due process of law in the practise of the Constitutional Court of Albania
Authors: Toska Dobjani Elsa
Published: January 2016

ECHR, as an international treaty is part of the Albanian legal system. Among international law instruments, the ECHR enjoys a privileged status in the Albanian legal system by virtue of Article 17 paragraph 2 of the Constitution according to which restrictions to human rights and freedoms cannot infringe the substance of those rights and freedoms and in no case can exceed the restrictions provided for in the ECHR. Article 1 of the Convention requires States to secure the substance of the rights to those in their jurisdiction. The effect of Article 13 is thus to require the provision of a domestic remedy to deal with the substance of an arguable complaint under the Convention and to grant appropriate relief. The scope of this paper is to analyse the effectiveness of the complaint to the Constitutional Court with regard to length of proceedings as part of due process of law in terms of proceedings during the court trial and after the process has been finalized and the final decision should be executed.
The effects of the global economic crisis on Macedonian economy: Some macroeconomic indicators and future policy recommendations
Authors: Tosheva Elizabeta
Published: January 2016

The recent economic and financial crisis caused a severe blow to the fragile Macedonian economy, halting the positive development trends in the last decade. This, has in turn, intensified the future challenges. The paper intends to analyze the impact of global economic crisis on the Macedonian economy focuses mainly on the macroeconomic level, identifying and analyzing fluctuations of major macroeconomic indicators that reflect the development and macroeconomic balances of the economy, such as GDP, the level of employment, inflation, budget deficit, public debt, etc. Secondary analysis of previous quantitative data and published studies, combined with an own qualitative study in the field, has provided a reliable and convincing basis for analysis. The experiences and lessons taken from the global economic crisis should serve as a basis for changing the current economic model with a new one in order the economy of the country to catch a connection with the intense changes that are expected to occur in the coming period. It is expected that creating new economic model in Republic of Macedonia will result in multiple positive effects that primarily manifested in the increasing number of newly small and medium enterprises, domestic investments, industrial production, GDP, number of new employees and total exports as well as in reduction of the trade deficit in maintaining macroeconomic stability of the country.
Sovereignity, statehood and self-determination in international law - The Kosovo case
Authors: Paço Spiro
Published: January 2016

The self-proclamation of Kosovo’s independence announced on February 17 invites us to some general considerations that may be linked to the specific event. The IL principle of Self-determination will be on the focus of this article to considering it as one of the way to archive the creation of new IL entity. Self-determination is a more recent principle then the others but its history, theory and practice is immense. The origin of the term and idea also is in discussion and debating among historian and lawyer. The term have been used simultaneously in the same period (end of WW I) by the US President Wilson and the USSR Premier Lenin referring to the right of the non self-governed territory like colony and occupied regions.
Review of “Marked Identities. Narrating lives between social labels and individual biographies”
Authors: Ferrarotti Laura
Published: January 2016

This is a book review of the volume edited by Roberta Piazza and Alessandra Fasulo, and entitled Marked Identities. Narrating Lives Between Social Labels and Individual Biographies, Palgrave Macmillan, London-New York, 2015. The book consists of various interviews with individuals whose lives, mainly because of the group to which they belong, could be viewed as ‘marked’. The difference between the idea of ‘stigma’ and the notion ‘marked identity’ is underlined, together with the idea of ‘diversity’, which today is not necessarily something that one should hide but, quite the opposite, something one can be proud of. One of the main ideas of this text is that identity is not a rigid reality but a process, by which individuals negotiate the version of who they are with others.
The acidic domain of the endothelial membrane protein GPIHBP1 stabilizes lipoprotein lipase activity by preventing unfolding of its catalytic domain
Authors: Mysling Simon , Kristensen Kristian Kølby , Larsson Mikael , Beigneux Anne P , Gårdsvoll Henrik , Fong Loren G , Bensadouen André , Jørgensen Thomas JD , Young Stephen G , Ploug Michael
Published: January 2016

GPIHBP1 is a glycolipid-anchored membrane protein of capillary endothelial cells that binds lipoprotein lipase (LPL) within the interstitial space and shuttles it to the capillary lumen. The LPL•GPIHBP1 complex is responsible for margination of triglyceride-rich lipoproteins along capillaries and their lipolytic processing. The current work conceptualizes a model for the GPIHBP1•LPL interaction based on biophysical measurements with hydrogen-deuterium exchange/mass spectrometry, surface plasmon resonance, and zero-length cross-linking. According to this model, GPIHBP1 comprises two functionally distinct domains: (1) an intrinsically disordered acidic N-terminal domain; and (2) a folded C-terminal domain that tethers GPIHBP1 to the cell membrane by glycosylphosphatidylinositol. We demonstrate that these domains serve different roles in regulating the kinetics of LPL binding. Importantly, the acidic domain stabilizes LPL catalytic activity by mitigating the global unfolding of LPL's catalytic domain. This study provides a conceptual framework for understanding intravascular lipolysis and GPIHBP1 and LPL mutations causing familial chylomicronemia.
Diffusion of hydroxyl groups in silica glass through the binding interface
Authors: Sato Naoya , Yamamoto Takaki , Kuzuu Nobu , Horikoshi Hideharu , Niwa Shohei
Published: January 2016

Diffusion of hydroxyl groups in silica glass through an interface formed by binding between high-hydroxyl (ca. 1200 wt.ppm) and low-hydroxyl (ca. 130 wt.ppm)-containing silica glasses in the temperature range of 900–1150 °C was investigated. Although the theoretical curve with a hydroxyl-concentration-independent diffusion coefficient deviates from the experimental curve, the diffusion coefficients obtained by fitting to the experimental results by the least squares method coincided with the “effective diffusion coefficients” in the literature, which were obtained from the total absorption change in the IR absorption peak for the hydroxyl group using thin samples. By the analysis considering the hydroxyl concentration dependence of the diffusion coefficient, we showed that the diffusion coefficient is proportional to hydroxyl concentration at each temperature, which is consistent with the model of the diffusion: SiOSi + H2O = 2SiOH. On the basis of this scheme, we tried to evaluate the diffusion coefficients of molecular water using equilibrium constant in the literature.
In Silico Functional Annotation of Genomic Variation
Authors: Butkiewicz Mariusz , Bush William S.
Published: January 2016

This unit describes the concepts and practical techniques for annotating genomic variants in the human genome to estimate their functional significance. With the rapid increase of available whole exome and whole genome sequencing information for human studies, annotation techniques have become progressively more important for highlighting and prioritizing nucleotide variants and their potential impact on genes and other genetic constructs. Here, we present an overview of different types of variant annotation approaches and elaborate on their foundations, assumptions, and the downstream consequences of their use. Computational approaches and tools to assign annotations and to identify variants are reviewed. Further, the general philosophy of assigning potential function to a genetic change within the biological context of a disease is discussed. © 2016 by John Wiley & Sons, Inc.
Spinal Muscular Atrophy: Overview of Molecular Diagnostic Approaches
Authors: Prior Thomas W. , Nagan Narasimhan
Published: January 2016

Spinal muscular atrophy (SMA) is an autosomal recessive neurodegenerative disease and the most common genetic cause of infant mortality, affecting ∼1 in 10,000 live births. The disease is characterized by progressive symmetrical muscle weakness resulting from the degeneration and loss of anterior horn cells in the spinal cord and brain stem nuclei. The disease is classified on the basis of age of onset and clinical course. SMA is caused by mutations in the telomeric copy of the survival motor neuron 1 (SMN1) gene, but all patients retain a centromeric copy of the gene, SMN2. The homozygous absence of the SMN1 exon 7 has been observed in the majority of patients and is being utilized as a reliable and sensitive SMA diagnostic test. In the majority of cases, the disease severity correlates inversely with an increased SMN2 gene copy number. Carrier detection, in the deletion cases, relies on the accurate determination of the SMN1 gene copies. Since SMA is one of the most common lethal genetic disorders, with a carrier frequency of 1 in 40 to 1 in 60, direct carrier dosage testing has been beneficial to many families. This unit attempts to highlight the molecular genetics of SMA with a focus on the advantages and limitations of the current molecular technologies. © 2016 by John Wiley & Sons, Inc.
Molecular Diagnosis of Cystic Fibrosis
Authors: Deignan Joshua L. , Grody Wayne W.
Published: January 2016

This unit describes a recommended approach to identifying causal genetic variants in an individual suspected of having cystic fibrosis. An introduction to the genetics and clinical presentation of cystic fibrosis is initially presented, followed by a description of the two main strategies used in the molecular diagnosis of cystic fibrosis: (1) an initial targeted variant panel used to detect only the most common cystic fibrosis‐causing variants in the CFTR gene, and (2) sequencing of the entire coding region of the CFTR gene to detect additional rare causal CFTR variants. Finally, the unit concludes with a discussion regarding the analytic and clinical validity of these approaches. © 2016 by John Wiley & Sons, Inc.
GONAD: A Novel CRISPR/Cas9 Genome Editing Method that Does Not Require Ex Vivo Handling of Embryos
Authors: Gurumurthy Channabasavaiah B. , Takahashi Gou , Wada Kenta , Miura Hiromi , Sato Masahiro , Ohtsuka Masato
Published: January 2016

Transgenic technologies used for creating a desired genomic change in animals involve three critical steps: isolation of fertilized eggs, microinjection of transgenic DNA into them and their subsequent transfer to recipient females. These ex vivo steps have been widely used for over 3 decades and they were also readily adapted for the latest genome editing technologies such as ZFNs, TALENs, and CRISPR/Cas9 systems. We recently developed a method called GONAD (Genome editing via Oviductal Nucleic Acids Delivery) that does not require all the three critical steps of transgenesis and therefore relieves the bottlenecks of widely used animal transgenic technologies. Here we provide protocols for the GONAD system. © 2016 by John Wiley & Sons, Inc.
Magnetic properties of sediments of the Red River: Effect of sorting on the source‐to‐sink pathway and its implications for environmental reconstruction
Authors: Nguyen Thi Thu Hien , Zhang Weiguo , Li Zhen , Li Jie , Ge Can , Liu Jinyan , Bai Xuexin , Feng Huan , Yu Lizhong
Published: January 2016

We conducted a mineral magnetic study of river bank and subaqueous delta sediments from the Red River, in order to examine the role of sedimentary sorting on the variation of sedimentary magnetic properties from source to sink. The magnetic mineralogy mainly consists of magnetite and hematite. Bulk sediment particle‐size variations have a strong influence on magnetic properties, with the frequently used magnetic parameters χfd%, χARM, χARM/χ, and χARM/SIRM exhibiting positive correlations with the <4 µm fraction, while S‐ratios are negatively correlated with this fraction. Compared with river bank sediments and shallow shoreface (<5 m water depth) sediments, sediments from the deeper (>5 m water depth) part of the subaqueous delta have lower χ and SIRM values, a finer ferrimagnetic grain‐size and higher proportions of hematite, consistent with selective loss of coarse ferrimagnetic grains on the source‐to‐sink pathway. We suggest that variations in magnetic properties in response to particle‐size compositions and therefore depositional environment changes should be carefully addressed when magnetic proxies such as χARM/SIRM are used in the study of coastal and marine environmental changes (e.g., sea‐level change). In such cases, the combined use of magnetic properties and geochemical indicators, such as Al/Ti ratio, may provide better results for paleoenvironmental reconstruction.
Combining AIRS and MLS observations for three‐dimensional gravity wave measurement
Authors: Wright Corwin J. , Hindley Neil P. , Mitchell Nicholas J.
Published: January 2016

Gravity waves play a critical role in transporting energy and momentum between the troposphere, stratosphere, and mesosphere. Satellite measurements provide a powerful tool to investigate these waves across the globe. However, many present methods cannot yield reliable estimates of wave momentum fluxes or the directions of these fluxes. Here we present a new method which addresses this problem by combining observations from Atmospheric Infrared Sounder (AIRS) and Microwave Limb Sounder (MLS) in three dimensions. The method allows direct estimation of horizontal and vertical wavelengths as well as wave amplitude. This in turn allows estimation of both wave momentum flux and the full 3‐D direction of propagation, crucially including the horizontal direction. The method thus allows separation of the data into, for example, eastward and westward momentum fluxes, allowing estimation of the net atmospheric forcing due to these waves. We illustrate this method with a proof‐of‐concept study over the Andes, arguably the largest source of gravity waves in the world. We further critically assess the advantages and disadvantages of our method. Our study highlights the importance of the difference between net and absolute measures of momentum flux.
The response of the equatorial ionosphere to fast stream solar coronal holes during 2008 deep solar minimum over Indian region
Authors: Sripathi S. , Singh Ram , Banola S. , Singh Dupinder , Sathish S.
Published: January 2016

In this paper, we report ionospheric response to fast stream solar coronal holes during 2008 deep solar minimum year using ground‐based multi‐instruments over Indian region. To examine this, we analyzed foF2 (MHz) and hpF2(km) from Canadian Advanced Digital Ionosonde and total electron content (TEC) from GPS receiver over Tirunelveli (8.73°N, 77.70°E; dip 0.5°N) along with equatorial electrojet (EEJ) strength. Our analysis shows good correlation between solar wind and foF2/TEC, while hpF2 is poorly correlated. However, moderate correlation exists between solar wind and EEJ strength. When we performed periodogram analysis, we observed 9 and 13 day periods as dominant periods in foF2 and TEC. Interestingly, the occurrence pattern of plasma irregularities also resembles these periodic oscillations. Since it is believed that lower atmospheric waves are dominant forces for ionospheric variabilities during deep solar minimum, we examined the mesosphere/lower thermosphere region temperature using Thermosphere Ionosphere Mesosphere Energetics and Dynamics Sounding of the Atmosphere using Broadband Emission Radiometry and winds using medium frequency radar along with outgoing longwave radiation in the troposphere altitudes to rule out the sources for these periodic oscillations in the lower atmosphere. Using cross‐wavelet and cross‐coherence spectra of both solar wind and ionospheric/atmospheric parameters, we suggest that ionospheric periodicities are similar to that of solar wind. Based on these results, we suggest that while the periodic oscillations are associated with the disturbance dynamo winds/electric fields that are propagated to equatorial latitudes, the differences in their temporal/seasonal variations are attributed to the variations in the composition/recombination changes.
Using orbital tethers to remediate geomagnetic radiation belts
Authors: Hudoba de Badyn Mathias , Marchand Richard , Sydora Richard D.
Published: January 2016

The Van Allen radiation belts pose a hazard to spacecraft and astronauts, and similar radiation belts around other planets pose a hazard to interplanetary probes. We discuss a method of remediating these radiation belts first proposed by Danilov and Vasilyev, and recently improved by Hoyt, Minor, and Cash, where a long, charged tether is placed in orbit inside a radiation belt. In this approach, an electric field of the tether scatters the belt particles into a pitch angle loss cone leading to absorption of the particles in the atmosphere. A test particle calculation is presented which computes the scattered pitch angle of belt particles as a function of initial pitch angle and gyrophase for different particle energies. The moments of the resulting distribution of scattered angle versus initial pitch angle are used to compute the number density of the belt as a function of time using a Fokker‐Planck diffusion approximation. Finally, we use the characteristic timescales of scattering for particles of different energies to discuss the feasibility of using such a system of tethers as a long and short‐term remediation solution.
Robust effects of cloud superparameterization on simulated daily rainfall intensity statistics across multiple versions of the Community Earth System Model
Authors: Kooperman Gabriel J. , Pritchard Michael S. , Burt Melissa A. , Branson Mark D. , Randall David A.
Published: January 2016

This study evaluates several important statistics of daily rainfall based on frequency and amount distributions as simulated by a global climate model whose precipitation does not depend on convective parameterization—Super‐Parameterized Community Atmosphere Model (SPCAM). Three superparameterized and conventional versions of CAM, coupled within the Community Earth System Model (CESM1 and CCSM4), are compared against two modern rainfall products (GPCP 1DD and TRMM 3B42) to discriminate robust effects of superparameterization that emerge across multiple versions. The geographic pattern of annual‐mean rainfall is mostly insensitive to superparameterization, with only slight improvements in the double‐ITCZ bias. However, unfolding intensity distributions reveal several improvements in the character of rainfall simulated by SPCAM. The rainfall rate that delivers the most accumulated rain (i.e., amount mode) is systematically too weak in all versions of CAM relative to TRMM 3B42 and does not improve with horizontal resolution. It is improved by superparameterization though, with higher modes in regions of tropical wave, Madden‐Julian Oscillation, and monsoon activity. Superparameterization produces better representations of extreme rates compared to TRMM 3B42, without sensitivity to horizontal resolution seen in CAM. SPCAM produces more dry days over land and fewer over the ocean. Updates to CAM's low cloud parameterizations have narrowed the frequency peak of light rain, converging toward SPCAM. Poleward of 50°, where more rainfall is produced by resolved‐scale processes in CAM, few differences discriminate the rainfall properties of the two models. These results are discussed in light of their implication for future rainfall changes in response to climate forcing.
Plant protein 2‐Cys peroxiredoxin TaBAS1 alleviates oxidative and nitrosative stresses incurred during cryopreservation of mammalian cells
Authors: Chow‐shi‐yée Mélanie , Grondin Mélanie , Averill‐Bates Diana A. , Ouellet François
Published: January 2016

There is increasing demand for cryopreserved cells such as liver and pancreatic cells for clinical applications. Cryopreservation at ultra‐low temperatures requires use of cryoprotectants (e.g., dimethyl sulfoxide (DMSO)) to maintain cell integrity during freezing and thawing processes. Standard cryoprotectants are cytotoxic and more effective cryopreservation technologies are urgently needed for long‐term storage of cells. As an alternative, soluble protein extracts (WPE) from winter wheat successfully replaced DMSO as a cryoprotectant for several mammalian cell types. To identify novel cryoactive proteins, the WPE was separated by chromatography and cryoactive fractions were analyzed by mass spectrometry. The wheat protein 2‐Cys peroxiredoxin BAS1 (renamed TaBAS1) was identified as a potential cryoactive candidate. Recombinant proteins were prepared and found to possess dual functions as a peroxidase antioxidant and molecular chaperone, and display cryoprotective properties for hepatocytes and insulin‐secreting INS832/13 cells. Following cryopreservation with TaBAS1, cells were plateable and showed high post‐thaw viability, good adhesion properties, and well‐maintained cell‐specific metabolic functions. The overall quality of these cell types was equivalent or improved compared to cells that were cryopreserved with DMSO. The antioxidant and chaperone functions of TaBAS1 likely explain its efficacy in reducing oxidative/nitrosative stresses in cryopreserved cells. The plant protein TaBAS1 could be a promising molecule to include in cryostorage protocols. Biotechnol. Bioeng. 2016;113: 1511–1521. © 2015 Wiley Periodicals, Inc.
Directed evolution increases desaturation of a cyanobacterial fatty acid desaturase in eukaryotic expression systems
Authors: Bai Shuangyi , Wallis James G. , Denolf Peter , Browse John
Published: January 2016

Directed evolution of a cyanobacterial Δ9 fatty acid desaturase (DSG) from Synechococcus elongatus, PCC6301 created new, more productive desaturases and revealed the importance of certain amino acid residues to increased desaturation. A codon‐optimized DSG open reading frame with an endoplasmic‐reticulum retention/retrieval signal appended was used as template for random mutagenesis. Increased desaturation was detected using a novel screen based on complementation of the unsaturated fatty acid auxotrophy of Saccharomyces cerevisiae mutant ole1Δ. Amino acid residues whose importance was discovered by the random processes were further examined by saturation mutation to determine the best amino acid at each identified location in the peptide chain and by combinatorial analysis. One frequently‐detected single amino acid change, Q240R, yielded a nearly 25‐fold increase in total desaturation in S. cerevisiae. Several other variants of the protein sequence with multiple amino acid changes increased total desaturation more than 60‐fold. Many changes leading to increased desaturation were in the vicinity of the canonical histidine‐rich regions known to be critical for electron transfer mediated by these di‐iron proteins. Expression of these evolved proteins in the seed of Arabidopsis thaliana altered the fatty acid composition, increasing monounsaturated fatty acids and decreasing the level of saturated fatty acid, suggesting a potential application of these desaturases in oilseed crops. Biotechnol. Bioeng. 2016;113: 1522–1530. © 2016 Wiley Periodicals, Inc.
Targeted transgene insertion into the CHO cell genome using Cre recombinase‐incorporating integrase‐defective retroviral vectors
Authors: Kawabe Yoshinori , Shimomura Takuya , Huang Shuohao , Imanishi Suguru , Ito Akira , Kamihira Masamichi
Published: January 2016

Retroviral vectors have served as efficient gene delivery tools in various biotechnology fields. However, viral DNA is randomly inserted into the genome, which can cause problems, such as insertional mutagenesis and gene silencing. Previously, we reported a site‐specific gene integration system, in which a transgene is integrated into a predetermined chromosomal locus of Chinese hamster ovary (CHO) cells using integrase‐defective retroviral vectors (IDRVs) and Cre recombinase. In this system, a Cre expression plasmid is transfected into founder cells before retroviral transduction. In practical applications of site‐specific gene modification such as for hard‐to‐transfect cells or for in vivo gene delivery, both the transgene and the Cre protein into retroviral virions should be encapsulate. Here, we generated novel hybrid IDRVs in which viral genome and enzymatically active Cre can be delivered (Cre‐IDRVs). Cre‐IDRVs encoding marker genes, neomycin resistance and enhanced green fluorescent protein (EGFP), flanked by wild‐type and mutated loxP sites were produced using an expression plasmid for a chimeric protein of Cre and retroviral gag‐pol. After analyzing the incorporation of the Cre protein into retroviral virions by Western blotting, the Cre‐IDRV was infected into founder CHO cells, in which marker genes (hygromycin resistance and red fluorescent protein) flanked with corresponding loxP sites are introduced into the genome. G418‐resistant colonies expressing GFP appeared and the site‐specific integration of the transgene into the expected chromosomal site was confirmed by PCR and sequencing of amplicons. Moreover, when Cre‐IDRV carried a gene expression unit for a recombinant antibody, the recombinant cells in which the antibody expression cassette was integrated in a site‐specific manner were generated and the cells produced the recombinant antibody. This method may provide a promising tool to perform site‐specific gene modification according to Cre‐based cell engineering. Biotechnol. Bioeng. 2016;113: 1600–1610. © 2016 Wiley Periodicals, Inc.