scholarly journals Comparative analysis of three studies measuring fluorescence from engineered bacterial genetic constructs

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0252263
Author(s):  
Jacob Beal ◽  
Geoff S. Baldwin ◽  
Natalie G. Farny ◽  
Markus Gershater ◽  
Traci Haddock-Angelli ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Comparative Analysis ◽  
Average Standard Deviation ◽  
The Third ◽  
Fluorescence Measurements ◽  
Interlaboratory Studies ◽  
Plate Reader ◽  
Order Of Magnitude ◽  
Bacterial Genetic ◽  
Genetic Constructs

Reproducibility is a key challenge of synthetic biology, but the foundation of reproducibility is only as solid as the reference materials it is built upon. Here we focus on the reproducibility of fluorescence measurements from bacteria transformed with engineered genetic constructs. This comparative analysis comprises three large interlaboratory studies using flow cytometry and plate readers, identical genetic constructs, and compatible unit calibration protocols. Across all three studies, we find similarly high precision in the calibrants used for plate readers. We also find that fluorescence measurements agree closely across the flow cytometry results and two years of plate reader results, with an average standard deviation of 1.52-fold, while the third year of plate reader results are consistently shifted by more than an order of magnitude, with an average shift of 28.9-fold. Analyzing possible sources of error indicates this shift is due to incorrect preparation of the fluorescein calibrant. These findings suggest that measuring fluorescence from engineered constructs is highly reproducible, but also that there is a critical need for access to quality controlled fluorescent calibrants for plate readers.

scholarly journals FlopR: An open source software package for calibration and normalization of plate reader and flow cytometry data

2020 ◽  
Author(s):  
Clare M. Robinson ◽  
Alex J. H. Fedorec ◽  
Ke Yan Wen ◽  
Chris P. Barnes
Keyword(s):  
Flow Cytometry ◽  
Synthetic Biology ◽  
Quantitative Measurement ◽  
Software Tool ◽  
Flow Cytometry Data ◽  
Statistical Framework ◽  
Fluorescence Measurements ◽  
Plate Reader ◽  
Open Source Software Package ◽  
Fluorescence Markers

AbstractThe measurement of gene expression using fluorescence markers has been a cornerstone of synthetic biology for the last two decades. However, the use of arbitrary units has limited the usefulness of this data for many quantitative purposes. Calibration of fluorescence measurements from flow cytometry and plate reader spectrophotometry has been implemented previously but the tools are disjointed. Here we pull together, and in some cases improve, extant methods into a single software tool, written as a package in the R statistical framework. The workflow is validated using Escherichia coli engineered to express GFP from a set of commonly used constitutive promoters. We then demonstrate its power by identifying the time evolution of distinct subpopulations of bacteria from bulk plate reader data, a task previously reliant on laborious flow cytometry experiments. Along with standardized parts and experimental methods, the development and dissemination of usable tools for quantitative measurement and data analysis will benefit the synthetic biology community by improving interoperability.Graphical Abstract

COMPARATIVE ANALYSIS OF THE GEOMETRICAL CONTENT IN THE NEW BULGARIAN TEXTBOOKS OF MATHEMATICS FOR THIRD GRADE

2018 ◽  
Vol 28 (3) ◽  
pp. 991-996
Author(s):  
Gabriela Kirova
Keyword(s):  
Comparative Analysis ◽  
Third Grade ◽  
Obtuse Angle ◽  
Curve Beam ◽  
Mathematics Textbook ◽  
The Third ◽  
School Year ◽  
Learning Content ◽  
Relative Share ◽  
Geometric Content

Starting with 2018/2019 school year in Bulgaria, the math education in the third grade is implemented through new training kits. They were developed on the basis of the new third-grade mathematics curriculum, approved by Order No. РД 09-1093 / 25.01.2017 of the Minister of Education and Science, Annex No. 8, supplemented by Order No. РД 09-2555 / 15.06.2018 of the Minister of Education and Science. Training kits are approved by the Ministry of Education and Science and are 7 in total. Geometric learning content in new math textbooks is the second most important element after arithmetic content. It is combined with the arithmetic learning content, and by this the foundation of the successful study of geometry in the next school grades is laid. The new geometry knowledge that is included in the third grade curriculum is the following: straight line, curve, beam, angle, right angle, obtuse angle, acute angle, right triangle, acute triangle, obtuse triangle; naming geometric figures with Latin alphabet letters [11]78. It is important in a modern mathematics textbook to have a rich and varied geometric content. It is important that the new types of geometry tasks are introduced with rich visualization using a specific-inductive approach. The relative number of tasks of a given type is an important prerequisite for the successful formation and improvement of skills for solving geometric problems in pupils at the age of 9-10. This article will present a comparative analysis of the geometric content in the seven approved Bulgarian third-grade mathematics textbooks, which are used in the mass practice of this school year. For the purpose of the study, a classification of all types of tasks and exercises with geometric content has been developed. Then the tasks in the seven textbooks are systematized by the so chosen classification. The data are statistically processed taking into account the relative share of tasks of each type within a textbook, as well as a comparison between the relative shares of the geometric tasks in the different textbooks. The established differences in the number and relative share of different types of geometric tasks make it possible for the analyzed textbooks to be ranked. Such a study has not been published so far. It has a relation to the assessment of the quality of the textbooks offered. The conclusions formulated in this article can help primary teachers in their choice of textbooks to teach to their third grade students.

scholarly journals Fluorescent Orthopalladated Complexes of 4-Aryliden-5(4H)-Oxazolones from the Kaede Protein: Synthesis and Characterization

Molecules ◽  
2021 ◽  
Vol 26 (5) ◽  
pp. 1238
Author(s):  
Eduardo Laga ◽  
David Dalmau ◽  
Sofía Arregui ◽  
Olga Crespo ◽  
Ana I. Jimenez ◽  
...  
Keyword(s):  
Intramolecular Interactions ◽  
Ortho Position ◽  
Fluorescent Properties ◽  
Ancillary Ligands ◽  
X Ray ◽  
Mononuclear Complexes ◽  
Fluorescence Measurements ◽  
Anionic Ligands ◽  
Order Of Magnitude

The goal of the work reported here was to amplify the fluorescent properties of 4-aryliden-5(4H)-oxazolones by suppression of the hula-twist non-radiative deactivation pathway. This aim was achieved by simultaneous bonding of a Pd center to the N atom of the heterocycle and the ortho carbon of the arylidene ring. Two different 4-((Z)-arylidene)-2-((E)-styryl)-5(4H)-oxazolones, the structures of which are closely related to the chromophore of the Kaede protein and substituted at the 2- and 4-positions of the arylidene ring (1a OMe; 1b F), were used as starting materials. Oxazolones 1a and 1b were reacted with Pd(OAc)2 to give the corresponding dinuclear orthometalated palladium derivates 2a and 2b by regioselective C–H activation of the ortho-position of the arylidene ring. Reaction of 2a (2b) with LiCl promoted the metathesis of the bridging carboxylate by chloride ligands to afford dinuclear 3a (3b). Mononuclear complexes containing the orthopalladated oxazolone and a variety of ancillary ligands (acetylacetonate (4a, 4b), hydroxyquinolinate (5a), aminoquinoline (6a), bipyridine (7a), phenanthroline (8a)) were prepared from 3a or 3b through metathesis of anionic ligands or substitution of neutral weakly bonded ligands. All species were fully characterized and the X-ray determination of the molecular structure of 7a was carried out. This structure has strongly distorted ligands due to intramolecular interactions. Fluorescence measurements showed an increase in the quantum yield (QY) by up to one order of magnitude on comparing the free oxazolone (QY < 1%) with the palladated oxazolone (QY = 12% for 6a). This fact shows that the coordination of the oxazolone to the palladium efficiently suppresses the hula-twist deactivation pathway.

Basil of Caesarea’s Authorship of Epistle 361 and His Relationship with the Homoiousians Reconsidered

2018 ◽  
Vol 72 (1) ◽  
pp. 41-70
Author(s):  
Dragoş Andrei Giulea
Keyword(s):  
Comparative Analysis ◽  
Third Century ◽  
Individual Substance ◽  
The Third ◽  
Mature Period ◽  
New Perspective ◽  
Eusebius Of Caesarea

Abstract A comparative analysis of Ep. 361 and Eun. 1.19 in terms of language and ideas will offer a renewed confirmation (on internal grounds) of Basil of Caesarea’s authorship of Ep. 361 and a new perspective on Basil’s relationship with the Homoiousians. In addition, the article will also retrace the steps and revisit the purpose of Basil’s argument. Thus we discover in the early Basil an author simultaneously receptive to both Homoiousian and pro-Nicene visions, but leaning towards an improved Homoiousian solution. The article further investigates Basil’s vision of ousia in Ep. 361 and finds that—unlike in his later, mature, period—the early Basil shares with the Homoiousians and Eusebius of Caesarea two doctrinal elements, namely the understanding of ousia as individual substance and an associated theology of “likeness”. He inherits this view from a tradition originating in the third century, which received its official confirmation at the council of Antioch in 268. This vision is also present in the first part of Basil’s Contra Eunomium. Instead of considering Basil as a Homoiousian, one may see him, together with Eusebius and the Homoiousians, as a representative of the Antiochene legacy.

scholarly journals Iran’s Oil and Gas Legal Framework: from Exploration to Revolution

2019 ◽  
Vol 6 (1) ◽  
pp. 15-22
Author(s):  
Mahmoud Fard Kardel
Keyword(s):  
Comparative Analysis ◽  
Turning Point ◽  
Oil And Gas ◽  
Oil And Gas Industry ◽  
Legal Framework ◽  
Gas Industry ◽  
The Third ◽  
Second Period ◽  
Petroleum Oil

The main purpose of this article is to examine Iran’s legal and contractual framework for their petroleum, oil and gas industry. Basically, the legal and contractual framework of the Iranian oil and gas industry has been classified into three periods. The first period is from the exploration and discovery of oil in Iran to nationalisation (1901-1951), the second period is from nationalisation to revolution (1951-1979), and the third period is from revolution to the present day (1979-20016).Because each period has its own features and importance two articles will examine this topic. The first period (from exploration to nationalisation, 1901-1951), and second period (from nationalisation to revolution, 1951-1979), has been examined in this article with legal and comparative analysis, and the third period (1979-2016) will be covered in a later article.It should be mentioned that each contractual framework was a turning point regarding to opportunities and circumstances that they have been in that time and also each of those petroleum contractual regimes were a step toward to contractual framework evolution in Iran.

scholarly journals The birth of minority legislation in the newly independent Finland and Estonia (1918–1920): a comparative analysis

2021 ◽  
Vol 13 (2) ◽  
pp. 61-84
Author(s):  
Kari Alenius
Keyword(s):  
Comparative Analysis ◽  
Political Parties ◽  
Interest Groups ◽  
Ethnic Minorities ◽  
Social Order ◽  
Historical Perspective ◽  
Cultural Rights ◽  
The Third ◽  
International Trends ◽  
The Relationship

This article analyzes how ethnic minorities were taken into account in the Finnish and Estonian constitutions, and why account was taken precisely in a certain way. At the same time, it approaches what kinds of views were presented by different political parties and interest groups, what kind of debate was being held in Parliament and how the matter was dealt with in the leading media. The outcome of the process in both countries was that exceptionally broad linguistic and cultural rights were given to minorities if the situation was compared with the rest of Europe. There were several factors behind the process. One factor was the relationship between ethnic groups in Finland and Estonia in the historical perspective. Another factor was each country's internal debate on what kind of social order in general was to be built. The third factor was how the politics in Finland and Estonia was influenced by international trends and theories about how ethnic minorities should have been treated.

scholarly journals Colorectal bradyarrhythmia syndrome as a predictor of metabolic syndrome

2020 ◽  
Vol 183 (11) ◽  
pp. 44-50
Author(s):  
K. A. Shemerovskii ◽  
V. N. Fedorets ◽  
P. V. Seliverstov ◽  
S. R. Bakaeva
Keyword(s):  
Metabolic Syndrome ◽  
Well Being ◽  
Medical Professionals ◽  
Early Prevention ◽  
The Third ◽  
The Metabolic Syndrome ◽  
Severe Stage ◽  
Order Of Magnitude ◽  
Two Stages

Evidence is presented for a significantly wider spread of the first two stages (mild and moderate) Colorectal Bradyarrhythmia Syndrome (CBS) compared with the third (severe — constipation) stage of this syndrome. Surveyed more than 2,500 medical professionals by the method of chronoenterographia. It was found that the incidence of mild CBS (about 60% of cases) and moderate stage of this syndrome (about 30% of cases) is almost an order of magnitude higher than the incidence of severe stage (about 10% of cases) of CBS. SCB has been shown to increase the risk of obesity by almost 3 times. It is shown that CBS contributes to a decrease in well-being, activity and mood, as well as a decrease in the quality of life. Screening for CBS and normalizing the circadian bowel rhythm with psyllium (mucofalc) in individuals who consider themselves healthy may contribute to early prevention of the risk of the metabolic syndrome.

scholarly journals CERAMIC ASSEMBLAGES OF THE THIRD QUARTER OF THE 15th CENTURY FROM THE FUNA CASTLE: UTENSILS OF THE «GARRISON KITCHEN»

2020 ◽  
Vol 34 (1) ◽  
pp. 31-67
Author(s):  
I. B. Teslenko
Keyword(s):  
Comparative Analysis ◽  
Historical Events ◽  
Archaeological Evidence ◽  
The Third ◽  
15Th Century ◽  
The Crimea ◽  
Three Stages ◽  
Last Stage ◽  
Ceramic Assemblages

The Funa fortress is located in southern Crimea and is one of the reference architectural and archaeological complexes of the Northern Pontic Region with precise date of existence. The fortress was built by Mangup authorities near 1423 on the border with possessions of Genoese and was destroyed in a fire during the Turkish invasion of the Crimea in 1475. The detailed chronology of the site which includes three stages of its construction history — 1423, 1425—1450s and 1459—1475, has been developed so far. So it becomes possible to clarify the dating of ceramic finds in line with these periods also. Ceramic assemblages of the last stage from the layers of fire and destruction of 1475 are the most representative. There is the complex from courtyard 1 among them. The ceramic collection includes 101 and 163 fully or partially reconstructed vessels respectively. There are large and average household containers, various kitchen utensils and tableware, both of the local Crimean production and import (Miletus Ware, Spanish Luster and Blue and White Ware, Fritware). The comparative analysis of artefacts made it possible to establish the chronological changes in ceramic assemblages during 25 years. Moreover, statistical and typological studies of the pottery from the layer of fire demonstrated a set of vessels there is suitable for cooking and table setting for at least 40 people. Large number of luxury tableware for diverse using and their location in the context allow suggest that there was a large feast on the platform above the «kitchen», and the remains of this banquet were not removed. According to the archaeological evidence as well as analysis of historical events the inhabitants of the fortress could burn it themselves before Turkish invasion and retreat to the capital of the principality at Mangup. Perhaps the remains of a farewell feast arranged just before leaving was fixed archaeologically.

Sign in / Sign up

Export Citation Format

Share Document