scholarly journals How to approach heterogeneous protein expression for biotechnological use: An overview

2017 ◽  
Vol 16 (1) ◽  
pp. 1-11 ◽  
Author(s):  
Daniela Jamrichová ◽  
Lenka Tišáková ◽  
Veronika Jarábková ◽  
Andrej Godány
Keyword(s):  
Protein Purification ◽  
Recombinant Proteins ◽  
Post Translational Modifications ◽  
Test Protein ◽  
Eukaryotic Gene ◽  
Common Problems ◽  
Escherichia Coli Expression ◽  
The Individual ◽  
Selection Of

AbstractProduction of recombinant proteins in Escherichia coli expression systems has shown many advantages, as well as disadvantages, especially for biotechnological and other down-stream applications. The choice of an appropriate vector depends on the gene, to be cloned for purification procedures and other analyses. Selection of a suitable production strain plays an important role in the preparation of recombinant proteins. The main criteria for the selection of the host organism are the properties of the recombinant produced protein, its subsequent use and the total amount desired. The most common problems in eukaryotic gene expression and recombinant proteins purification are, for instance, post-translational modifications, formation of disulphide bonds, or inclusion bodies. Obtaining a purified protein is a key step enabling further characterization of its role in the biological system. Moreover, methods of protein purification have been developed in parallel with the discovery of proteins and the need for their studies and applications. After protein purification, and also between the individual purification steps, it is necessary to test protein stability under different conditions over time. Shortly, all the essential points have been briefly discussed, which could be encountered during production and purification of a recombinant protein of interest, especially from eukaryotic source and expressed heterogeneously in prokaryotic production system.

scholarly journals Preclinical Testing of Radiopharmaceuticals for the Detection and Characterization of Osteomyelitis: Experiences from a Porcine Model

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4221
Author(s):  
Aage Kristian Olsen Alstrup ◽  
Svend Borup Jensen ◽  
Ole Lerberg Nielsen ◽  
Lars Jødal ◽  
Pia Afzelius
Keyword(s):  
Animal Model ◽  
Animal Models ◽  
Porcine Model ◽  
Animal Experiments ◽  
Radioactive Tracers ◽  
The Individual ◽  
Selection Of

The development of new and better radioactive tracers capable of detecting and characterizing osteomyelitis is an ongoing process, mainly because available tracers lack selectivity towards osteomyelitis. An integrated part of developing new tracers is the performance of in vivo tests using appropriate animal models. The available animal models for osteomyelitis are also far from ideal. Therefore, developing improved animal osteomyelitis models is as important as developing new radioactive tracers. We recently published a review on radioactive tracers. In this review, we only present and discuss osteomyelitis models. Three ethical aspects (3R) are essential when exposing experimental animals to infections. Thus, we should perform experiments in vitro rather than in vivo (Replacement), use as few animals as possible (Reduction), and impose as little pain on the animal as possible (Refinement). The gain for humans should by far exceed the disadvantages for the individual experimental animal. To this end, the translational value of animal experiments is crucial. We therefore need a robust and well-characterized animal model to evaluate new osteomyelitis tracers to be sure that unpredicted variation in the animal model does not lead to a misinterpretation of the tracer behavior. In this review, we focus on how the development of radioactive tracers relies heavily on the selection of a reliable animal model, and we base the discussions on our own experience with a porcine model.

Amplification of Signal Features Using Variance Fractal Dimension Trajectory

2010 ◽  
Vol 4 (4) ◽  
pp. 1-17 ◽  
Author(s):  
Witold Kinsner ◽  
Warren Grieder
Keyword(s):  
Fractal Dimension ◽  
Time Domain ◽  
Speech Signal ◽  
Ambient Noise ◽  
Unit Distance ◽  
Detection Techniques ◽  
Signal Features ◽  
The Individual ◽  
Selection Of

This paper describes how the selection of parameters for the variance fractal dimension (VFD) multiscale time-domain algorithm can create an amplification of the fractal dimension trajectory that is obtained for a natural-speech waveform in the presence of ambient noise. The technique is based on the variance fractal dimension trajectory (VFDT) algorithm that is used not only to detect the external boundaries of an utterance, but also its internal pauses representing the unvoiced speech. The VFDT algorithm can also amplify internal features of phonemes. This fractal feature amplification is accomplished when the time increments are selected in a dyadic manner rather than selecting the increments in a unit distance sequence. These amplified trajectories for different phonemes are more distinct, thus providing a better characterization of the individual segments in the speech signal. This approach is superior to other energy-based boundary-detection techniques. Observations are based on extensive experimental results on speech utterances digitized at 44.1 kilosamples per second, with 16 bits in each sample.

scholarly journals Development of an ELISA-Based HDAC Activity Assay for Characterization of Isoform-Selective Inhibitors

2015 ◽  
Vol 20 (10) ◽  
pp. 1277-1285 ◽  
Author(s):  
Geetha Padige ◽  
Ahmed T. Negmeldin ◽  
Mary Kay H. Pflum
Keyword(s):  
Mammalian Cell ◽  
Recombinant Proteins ◽  
Hdac Inhibitors ◽  
Enzyme Linked Immunosorbent Assay ◽  
Activity Assay ◽  
Hdac Activity ◽  
Anticancer Properties ◽  
The Individual ◽  
Hdac Proteins

Histone deacetylase (HDAC) proteins are promising targets for cancer treatment, with several HDAC inhibitors used clinically as anticancer drugs. Most HDAC inhibitors nonspecifically interact with all or many of the 11 HDAC isoforms. Isoform-selective HDAC inhibitors would be useful tools to dissect the individual functions of HDAC proteins in cancer formation, in addition to potentially displaying effective anticancer properties. We report here a robust HDAC activity assay for screening selective HDAC inhibitors, which is inspired by the traditional enzyme-linked immunosorbent assay (ELISA). The key feature of the ELISA-based HDAC activity assay is use of mammalian cell–derived HDAC isoforms instead of recombinant proteins. Importantly, the assay was validated with several known HDAC inhibitors. The ELISA-based HDAC activity assay will facilitate the characterization of isoform-selective HDAC inhibitors against mammalian cell–derived HDAC proteins, which will enhance HDAC-centered cancer research and provide a foundation for anticancer drug development.

scholarly journals FUNKCIJA SELEKCIJE, KARAKTERIZACIJE I KONSTELACIJE LIKOVA U ROMANU VOLFGANGA HERNDORFA „ČIK“

2021 ◽  
Vol 18 (49) ◽  
pp. 303-320
Author(s):  
Ivana Pajić ◽  
Keyword(s):  
Developmental Process ◽  
Methodological Approach ◽  
The Novel ◽  
Social Image ◽  
Award Winning ◽  
Self Development ◽  
The Subject ◽  
The Individual ◽  
Selection Of

The subject of the present work is Wolfgang Herrndorf’s award-winning youth novel Tsch- ick (i.e. “German Youth Literature Prize” (2011) and “Clemens Brentano Prize” (2012)). The novel, released in 2010, has sold more than two million copies to date and has been translated into more than 25 languages. In the novel, the story is told by Maik Klingenberg, a fourteen- year-old teenager who is left devastated by the surrounding circumstances and his outsider position before the summer holidays, but through his friendship with Tschick (during the summer holidays) experiences a solution and development process that changes his view of the world and his positioning to his environment and to himself in a positive way. The main focus of the work lies on the (explicit and implicit) character selection, characterization and constellation in the text. The methodological approach to textual analysis is based on insights from narratology, sociology and psychoanalysis. After a theoretical-introductory part, the text-analytical part begins with the selection of the staff of the fictitious world depicted in the novel and shows that the author’s novel gives a pluralistic-heterogeneous-inter- or transcul- tural social image of the German capital Berlin and its (wider) environment. In the following chapter of the work, the introductory characterization of the main character and the related figure constellations will be discussed in more detail. It is shown that Herrndorf uses these constellations to address the complex interrelation between the individual and various col- lective identities, such as those into which the subject is born (Maik’s parents), and those to whom the subject belongs based on the present circumstances of life (Maik’s teacher and the other students of the school). The focus of analysis of the following part lies on those figure constellations that have arisen as a result of Maik’s escape from the structures crushing him (Maik’s voluntary social interactions and friendly ties) and by which the author refers to the dynamic constitution of personal identity, of which the developmental process defies static defining and reveals the possibility of self-change and self-development. Based on the pre- viously given text analysis, the last part of the thesis gives a conclusion corresponding to the analysis results.

Amplification of Signal Features Using Variance Fractal Dimension Trajectory

2012 ◽  
pp. 263-277
Author(s):  
Witold Kinsner ◽  
Warren Grieder
Keyword(s):  
Fractal Dimension ◽  
Time Domain ◽  
Speech Signal ◽  
Experimental Results ◽  
Unit Distance ◽  
Detection Techniques ◽  
Signal Features ◽  
The Individual ◽  
Selection Of

This paper describes how the selection of parameters for the variance fractal dimension (VFD) multiscale time-domain algorithm can create an amplification of the fractal dimension trajectory that is obtained for a natural-speech waveform in the presence of ambient noise. The technique is based on the variance fractal dimension trajectory (VFDT) algorithm that is used not only to detect the external boundaries of an utterance, but also its internal pauses representing the unvoiced speech. The VFDT algorithm can also amplify internal features of phonemes. This fractal feature amplification is accomplished when the time increments are selected in a dyadic manner rather than selecting the increments in a unit distance sequence. These amplified trajectories for different phonemes are more distinct, thus providing a better characterization of the individual segments in the speech signal. This approach is superior to other energy-based boundary-detection techniques. Observations are based on extensive experimental results on speech utterances digitized at 44.1 kilosamples per second, with 16 bits in each sample.

New developments in post-translational modifications and functions of histone H2A variantsThis paper is one of a selection of papers published in this Special Issue, entitled CSBMCB’s 51st Annual Meeting – Epigenetics and Chromatin Dynamics, and has undergone the Journal’s usual peer review process.

2009 ◽  
Vol 87 (1) ◽  
pp. 7-17 ◽  
Author(s):  
Anita A. Thambirajah ◽  
Andra Li ◽  
Toyotaka Ishibashi ◽  
Juan Ausió
Keyword(s):  
Review Process ◽  
Peer Review Process ◽  
Histone Variants ◽  
Histone H2a ◽  
Post Translational Modifications ◽  
New Developments ◽  
Chromatin Dynamics ◽  
Recent Developments ◽  
The Individual ◽  
Selection Of

Structural variability within histone families, such as H2A, can be achieved through 2 primary mechanisms: the expression of histone variants and the incorporation of chemical modifications. The histone H2A family contains several variants in addition to the canonical H2A forms. In this review, recent developments in the study of the heteromorphous variants H2A.X, H2A.Z, and macroH2A will be discussed. Particular focus will be given to the post-translational modifications (PTMs) of these variants, including phosphorylation, ubiquitination, acetylation, and methylation. The combination of the newly identified N- and C-terminal tail PTMs expands the multiplicity of roles that the individual H2A variants can perform. It is of additional interest that analogous sites within these different histone variants can be similarly modified. Whether this is a redundant function or a finely tuned one, designed to meet specific needs, remains to be elucidated.

Applications of Transmission Electron Microscopy in the Characterization of Metal Machinability

1968 ◽  
Vol 26 ◽  
pp. 442-443 ◽  
Author(s):  
L.E. Murr ◽  
A.B. Draper
Keyword(s):  
Electron Microscopy ◽  
State Physics ◽  
Test Material ◽  
Milling Machine ◽  
Rotary Table ◽  
Solid State Physics ◽  
Materials Properties ◽  
Transmission Electron ◽  
Selection Of

The industrial characterization of the machinability of metals and alloys has always been a very arbitrarily defined property, subject to the selection of various reference or test materials; and the adoption of rather naive and misleading interpretations and standards. However, it seems reasonable to assume that with the present state of knowledge of materials properties, and the current theories of solid state physics, more basic guidelines for machinability characterization might be established on the basis of the residual machined microstructures. This approach was originally pursued by Draper; and our presentation here will simply reflect an exposition and extension of this research.The technique consists initially in the production of machined chips of a desired test material on a horizontal milling machine with the workpiece (specimen) mounted on a rotary table vice. A single cut of a specified depth is taken from the workpiece (0.25 in. wide) each at a new tool location.

EVALUATION OF MECHANICAL SYSTEMS USED IN PERFUSION

1972 ◽  
Vol 68 (2_Supplb) ◽  
pp. S44-S73 ◽  
Author(s):  
Eugene F. Bernstein
Keyword(s):  
Mechanical Systems ◽  
Organ Perfusion ◽  
Critical Factors ◽  
System 2 ◽  
Common Problems ◽  
Mechanical Components ◽  
Perfusion Experiment ◽  
Selection Of

ABSTRACT Among the critical factors in organ perfusion are (1) the mechanical components of the system, (2) the composition of the perfusate, and (3) the perfusing conditions. In this review, particular consideration is given to the pump, the oxygenator, and cannulas in such systems. Emphasis is placed upon the selection of pertinent equipment for the goals of a particular perfusion experiment, based upon the criteria of adequacy of the perfusion. Common problems in organ perfusion are summarized, and potential solutions to the perfusion problem, involving either biologic or mechanical extracorporeal systems, are suggested.

Pengaruh Persepsi Kegunaan, Persepsi Kemudahan, Kecepatan, Keamanan 75 dan Kerahasiaan Serta Kesiapan Teknologi Informasi Wajib Pajak terhadap Penggunaan e-Filing Bagi Wajib Pajak Orang Pribadi Di Kota Tangerang, Kecamatan Karawaci

2016 ◽  
Vol 8 (1) ◽  
pp. 75-91 ◽  
Author(s):  
Shelby Devina ◽  
Waluyo Waluyo
Keyword(s):  
Ease Of Use ◽  
Perceived Usefulness ◽  
Security And Privacy ◽  
Primary Data ◽  
Perceived Ease Of Use ◽  
Analysis Technique ◽  
Convenience Sampling ◽  
The Individual ◽  
Tax Payers ◽  
Selection Of

The objective of this research was to examine the effect of perceived usefulness, perceived ease of use, speed, security and privacy and readiness technology tax payers information to e-Filing usage. The object of this study is the individual tax payers in Tangerang City, Karawaci District. The selection of the sample is determined based on convenience sampling method. Data used in this study was primary data, id est: questionnaires. The respondent in this study were 110. Data analysis technique in this study using multiple linear regression. The result of this study were (1) perceived usefulness have a significant impact towards e-Filing usage; (2) perceived ease of use have a significant impact towards e-Filing usage; (3) speed does not have a significant impact towards e-Filing usage; (4) security and privacy does not have a significant impact towards e-Filing usage; (5) readiness technology tax payers information does not have a significant impact towards e-Filing usage; (6) perceived usefulness, perceived ease of use, speed, security and privacy and readiness technology tax payers information all simultaneously, have a significant impact towards e-Filing usage. Keywords: e-Filing usage, perceived usefulness, perceived ease of use, readiness technology tax payers information, security and privacy.

Caracterização da Peneira Média em Clones de Coffea canephora

REVISTA FIMCA ◽  
2018 ◽  
Vol 5 (2) ◽  
pp. 28-31
Author(s):  
Darlan Darlan Sanches Barbosa Alves ◽  
Victor Mouzinho Spinelli ◽  
Marcos Santana Moraes ◽  
Carolina Augusto De Souza ◽  
Rodrigo da Silva Ribeiro ◽  
...  
Keyword(s):  
Genetic Improvement ◽  
Block Design ◽  
Coffea Canephora ◽  
Coffee Production ◽  
Randomized Block Design ◽  
Significant Difference ◽  
The Mean ◽  
Four Blocks ◽  
Selection Of

Introdução: O estado de Rondônia se destaca como tradicional produtor de café, sendo o segundo maior produtor brasileiro de C. canephora. No melhoramento genético de C. canephora, a seleção de plantas de elevada peneira média está associada à bebida de qualidade superior. Objetivos: O objetivo desse estudo foi avaliar a variabilidade genética de clones de C. canephora para o tamanho dos grãos, mensurado a partir da avaliação da peneira média (PM). Materiais e Métodos: Para isso, foi conduzido ao longo de dois anos agrícolas experimento no campo experimental da Embrapa no município de Ouro Preto do Oeste-RO, para a avaliação da peneira média de 130 genótipos (clones) com características das variedades botânicas Conilon, Robusta e híbridos intervarietais. O delineamento experimental utilizado foi de blocos ao acaso, com quatro repetições de quatro plantas por parcela. Resultados: Não houve resultados significativos para a interação clones X anos, indicando uma maior consistência no comportamento das plantas ao longo do tempo. Porém foram observadas diferenças significativas para o tamanho dos grãos entre os genótipos avaliados, possibilitando selecionar genótipos superiores. Conclusão: Os genótipos agruparam-se em cinco classes de acordo com o teste de média, subsidiando a caracterização de um gradiente de variabilidade da característica avaliada ABSTRACTIntroduction: Coffea canephora accounts for approximately 35% of the world's coffee production. The state of Rondônia stands out as a traditional coffee producer, being the second largest Brazilian producer of C. canephora. In the classical genetic improvement of C. anephora, the selection of plants of high average sieve is associated with a drink of superior quality. Objectives: The objective of this udy was to evaluate the genetic variability of Coffea canephora clones for the agronomic medium sieve (PM). Materials and Methods: The experiment was conducted in the experimental field of Embrapa, municipality of OuroPreto do Oeste-RO, located at coordinates 10º44'53 "S and 62º12'57". One hundred thirty genotypes (clones) of botanical characteristics Conilon, Robusta and intervarietal hybrids were evaluated in the agricultural years 2013-2014 and 2014-2015. The experimental design was a randomized block design with four blocks and four plants per plot, spacing 3.5 x 1.5 meters between plants. Results: Significant difference was found for the grain size. According to the F test, at 5% probability, the genotypes were grouped into five classes according to the mean test. Conclusion: The results obtained subsidized the characterization of a variability gradient of the evaluated trait.

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