scholarly journals Machine-driven parameter screen of biochemical reactions

2020 ◽  
Vol 48 (7) ◽  
pp. e37-e37
Author(s):  
Stéphane Poulain ◽  
Ophélie Arnaud ◽  
Sachi Kato ◽  
Iris Chen ◽  
Hiro Ishida ◽  
...  
Keyword(s):  
High Performance ◽  
Transfer Programs ◽  
Methods Development ◽  
Biochemical Systems ◽  
Liquid Transfer ◽  
Parameter Interactions ◽  
The Cost ◽  
Computer Generation ◽  
Multiwell Plates

Abstract The development of complex methods in molecular biology is a laborious, costly, iterative and often intuition-bound process where optima are sought in a multidimensional parameter space through step-by-step optimizations. The difficulty of miniaturizing reactions under the microliter volumes usually handled in multiwell plates by robots, plus the cost of the experiments, limit the number of parameters and the dynamic ranges that can be explored. Nevertheless, because of non-linearities of the response of biochemical systems to their reagent concentrations, broad dynamic ranges are necessary. Here we use a high-performance nanoliter handling platform and computer generation of liquid transfer programs to explore in quadruplicates 648 combinations of 4 parameters of a biochemical reaction, the reverse-transcription, which lead us to uncover non-linear responses, parameter interactions and novel mechanistic insights. With the increased availability of computer-driven laboratory platforms for biotechnology, our results demonstrate the feasibility and advantage of methods development based on reproducible, computer-aided exhaustive characterization of biochemical systems.

scholarly journals Machine-driven parameter optimisation of biochemical reactions

2019 ◽  
Author(s):  
Stéphane Poulain ◽  
Ophélie Arnaud ◽  
Sachi Kato ◽  
Iris Chen ◽  
Hiro Ishida ◽  
...  
Keyword(s):  
High Performance ◽  
Transfer Programs ◽  
Methods Development ◽  
Biochemical Systems ◽  
Liquid Transfer ◽  
Parameter Interactions ◽  
The Cost ◽  
Computer Generation ◽  
Multi Step Methods ◽  
Multiwell Plates

AbstractThe development of complex, multi-step methods in molecular biology is a laborious, costly, iterative and often intuition-bound process where an optimum is sought in a multidimensional parameter space through step-by-step optimisations. The difficulty of miniaturising reactions under the microliter volumes usually handled in multiwell plates by robots, plus the cost of the experiments, limit the number of parameters and the dynamic ranges that can be explored. Nevertheless, because of non-linearities of the response of biochemical systems to their reagent concentrations, broad dynamic ranges are necessary. Here we use a high-performance nanoliter handling platform and computer generation of liquid transfer programs to explore in quadruplicates more than 600 combinations of 4 parameters of a biochemical reaction, the reverse-transcription, which lead us to uncover non-linear responses, parameter interactions and novel mechanistic insights. With the increased availability of computer-driven laboratory platforms for biotechnology, our results demonstrate the feasibility and advantage of methods development based on reproducible, computer-aided exhaustive characterisation of biochemical systems.

scholarly journals Natural Dyes from Mortiño (Vaccinium floribundum) as Sensitizers in Solar Cells

Energies ◽  
2020 ◽  
Vol 13 (4) ◽  
pp. 785 ◽  
Author(s):  
Miguel A. Taco-Ugsha ◽  
Cristian P. Santacruz ◽  
Patricio J. Espinoza-Montero
Keyword(s):  
High Performance Liquid Chromatography ◽  
Solar Cells ◽  
Liquid Chromatography ◽  
Solar Cell ◽  
High Performance ◽  
Power Conversion ◽  
Natural Dyes ◽  
The Cost ◽  
Layer Chromatography

Photovoltaic energy presents environmental advantages; however, these advantages are limited by the cost of manufacturing solar cells and in many cases, scarce or dangerous materials are incorporated. Therefore, the use of natural dyes from mortiño (Vaccinium floribundum) as sensitizers in solar cells is proposed. The dyes were extracted by maceration in acidified methanol (HCl, citric acid and trifluoroacetic acid TFA) and were characterized by High-Performance Liquid Chromatography (HPLC), Thin-Layer Chromatography (TLC) and spectrometric methods (UV-Vis, IR and MS-MALDI). The construction and characterization of cells were in standard conditions. The study confirms that pigments in mortiño are flavonoids of the anthocyanidin group as: cyanidin-3-galactoside, and cyanidin-3-arabinoside. The efficiency of solar cells was between 0.18–0.26%; the extraction with TFA in methanol leads to the best performance. Although they have low power conversion efficiency, mortiño dyes could be an alternative to artificial sensitizers for solar cell technologies because they are harmless and abundant substances.

Preparation and Characterization of NH2-Terminal Fibrinogen Bβ Fragments from N-DSK of Human Fibrinogen

1984 ◽  
Vol 51 (01) ◽  
pp. 016-021 ◽  
Author(s):  
S Birken ◽  
G Agosto ◽  
B Lahiri ◽  
R Canfield
Keyword(s):  
High Performance ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Reverse Phase ◽  
Human Fibrinogen ◽  
Human Volunteers ◽  
Cnbr Cleavage ◽  
Two Peaks

SummaryIn order to investigate the early release of NH2-terminal plasmic fragments from the Bβ chain of fibrinogen, substantial quantities of Bβ 1-42 and Bβ 1-21 are required as immunogens, as radioimmunoassay standards and for infusion into human volunteers to determine the half-lives of these peptides. Towards this end methods that employ selective proteolytic cleavage of these fragments from fibrinogen have been developed. Both the N-DSK fragment, produced by CNBr cleavage of fibrinogen, and Bβ 1-118 were employed as substrates for plasmin with the finding of higher yields from N-DSK. Bβ 1-42 and Bβ 1-21 were purified by gel filtration and ion-exchange chromatography on SP-Sephadex using volatile buffers. When the purified preparation of Bβ 1-42 was chromatographed on reverse-phase high performance liquid chromatography, two peaks of identical amino acid composition were separated, presumably due either to pyroglutamate or to amide differences.

Preparation and Characterization of the High Molecular Weight [3H]Hyaluronic Acid

1992 ◽  
Vol 57 (10) ◽  
pp. 2151-2156 ◽  
Author(s):  
Peter Chabreček ◽  
Ladislav Šoltés ◽  
Hynek Hradec ◽  
Jiří Filip ◽  
Eduard Orviský
Keyword(s):  
Molecular Weight ◽  
Hyaluronic Acid ◽  
High Molecular Weight ◽  
Methyl Bromide ◽  
High Performance ◽  
Hydrogen Atoms ◽  
Isolation And Characterization ◽  
Labelling Procedure ◽  
Enzymatic Depolymerization

Two methods for the preparation of high molecular weight [3H]hyaluronic acid were investigated. In the first one, hydrogen atoms in the molecule were replaced by tritium. This isotopic substitution was performed in aqueous solution using Pd/CaCO3 as the catalyst. In the second method, the high molecular weight hyaluronic acid was alkylated with [3H]methyl bromide in liquid ammonia at a temperature of -33.5 °C. High-performance gel permeation chromatographic separation method was used for the isolation and characterization of the high molecular weight [3H]hyaluronic acid. Molecular weight parameters for the labelled biopolymers were Mw = 128 kDa, Mw/Mn = 1.88 (first method) and Mw = 268 kDa, Mw/Mn = 1.55 (second method). The high molecular weight [3H]hyaluronic acid having Mw = 268 kDa was degraded further by specific hyaluronidase. Products of the enzymatic depolymerization were observed to be identical for both, labelled and cold biopolymer. This finding indicates that the described labelling procedure using [3H]methyl bromide does not induce any major structural rearrangements in the molecule.

scholarly journals AB1062 LIPODERMATOSCLEROSIS AS A TYPE OF LOBULAR PANNICULITIS: THE EFFECTIVENESS OF NON-PHARMACOLOGICAL TREATMENT METHODS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1821.1-1821
Author(s):  
M. Sukhareva ◽  
O. Egorova ◽  
B. Belov
Keyword(s):  
Subcutaneous Fat ◽  
Lymphatic Drainage ◽  
Normal Body Mass Index ◽  
Positive Trend ◽  
Group I ◽  
Positive Clinical Effect ◽  
Group Ii ◽  
The Cost ◽  
Lobular Panniculitis

Background:In medical practice lobular panniculitis-lipodermatosclerosis (LDS) is becoming more and more common. It is manifested by degenerative-dystrophic changes in subcutaneous fat (SCF) and occurs more often in middle-aged women affected by chronic venous insufficiency.Objectives:to evaluate the effectiveness of mesotherapy (MT) and shockwave ultrasound therapy (UST) for LDSMethods:among 539 patients referred to the V.A. Nasonova Research Institute of Rheumatology with the referral diagnoses of erythema nodosum or panniculitis 8.5% (46) of patients (44 women, 2 men) aged 18 to 82 with overweight (32) LDS with the disease duration of 11,8±6.4 months was verified. Patients were randomized into two groups of 23 patients each: group I received daily MT (10 sessions) therapy with drugs that have antioxidant, anti-inflammatory, lymphatic drainage and lipolytic effects, and 3 MHz UST of the node area twice a week (5 sessions). In group II MT was performed daily with 9% Natrii chloridum solution at a dose comparable to group I. The control methods included general clinical examination (characterization of induration on the lower legs with an assessment of the effect of pain pressing according to visual analogue scale (VAS pain), general blood and urine tests and ultrasound with elastography (USE) of the compaction. The main stages of control: initial (T0), after 14 days (T1), 1 month (T2) and 3 months (T3).Results:before treatment 38 patients with LDS demonstrated asymmetric (83%) inflammation of SCF of the lower legs (100%) on its medial surface (91%). LDS regressed faster with normal body mass index (p = 0,04). In all patients of group I, after a course of physiotherapy a positive trend was registered, that is a decrease in VAS pain intensity (T0 50±18 mm; T1 35±11 mm), decrease in diameter (T0 6±2.2 cm; T1 4.5±1, 7 mm) and color intensity of the node (p<0.002), SCF thickening which results in “lumping” with macrovascularization according to USE, and decrease in ESR and CRP. In 44% of cases the treatment effect increased to T2 (p <0.05). After 3 months of observation, 15 patients required a second course of physiotherapy. In group II a positive clinical effect was registered for T2 in 14 patients (60.8%) and for T3 in 19 patients (83%) (p<0.05). Over the entire observation period LDS recurrence was registered in 19 patients (41%), the median of recurrence was 3 [1; 6] months, mainly in patients of group I. Recurrence was associated with node fusion into conglomerates (OR 4.33, 95% CI 1.05-17.8; p = 0.037). MT and UST were tolerated well, no side effects were detected.Conclusion:the use of MT with 9% Natrii chloridum solution allowed us to achieve positive dynamics in patients with LDS, which significantly reduced the cost of treatment. Further studies are needed to evaluate the significance of these techniques.Disclosure of Interests:None declared

scholarly journals Development and characterization of Nb3Sn/Al2O3 superconducting multilayers for particle accelerators

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chris Sundahl ◽  
Junki Makita ◽  
Paul B. Welander ◽  
Yi-Feng Su ◽  
Fumitake Kametani ◽  
...  
Keyword(s):  
High Performance ◽  
High Energy ◽  
Particle Accelerators ◽  
Quality Factors ◽  
Critical Fields ◽  
Linear Accelerators ◽  
Cross Sectional ◽  
Low Field ◽  
Superconducting Radio Frequency

AbstractSuperconducting radio-frequency (SRF) resonator cavities provide extremely high quality factors > 1010 at 1–2 GHz and 2 K in large linear accelerators of high-energy particles. The maximum accelerating field of SRF cavities is limited by penetration of vortices into the superconductor. Present state-of-the-art Nb cavities can withstand up to 50 MV/m accelerating gradients and magnetic fields of 200–240 mT which destroy the low-dissipative Meissner state. Achieving higher accelerating gradients requires superconductors with higher thermodynamic critical fields, of which Nb3Sn has emerged as a leading material for the next generation accelerators. To overcome the problem of low vortex penetration field in Nb3Sn, it has been proposed to coat Nb cavities with thin film Nb3Sn multilayers with dielectric interlayers. Here, we report the growth and multi-technique characterization of stoichiometric Nb3Sn/Al2O3 multilayers with good superconducting and RF properties. We developed an adsorption-controlled growth process by co-sputtering Nb and Sn at high temperatures with a high overpressure of Sn. The cross-sectional scanning electron transmission microscope images show no interdiffusion between Al2O3 and Nb3Sn. Low-field RF measurements suggest that our multilayers have quality factor comparable with cavity-grade Nb at 4.2 K. These results provide a materials platform for the development and optimization of high-performance SIS multilayers which could overcome the intrinsic limits of the Nb cavity technology.

scholarly journals Enzymatic Synthesis and Characterization of Different Families of Chitooligosaccharides and Their Bioactive Properties

2021 ◽  
Vol 11 (7) ◽  
pp. 3212
Author(s):  
Noa Miguez ◽  
Peter Kidibule ◽  
Paloma Santos-Moriano ◽  
Antonio O. Ballesteros ◽  
Maria Fernandez-Lobato ◽  
...  
Keyword(s):  
High Performance ◽  
Chemical Characterization ◽  
Amperometric Detection ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Enzymatic Production ◽  
Bioactive Properties ◽  
Substrate Properties ◽  
Chitin Hydrolysis

Chitooligosaccharides (COS) are homo- or hetero-oligomers of D-glucosamine (GlcN) and N-acetyl-D-glucosamine (GlcNAc) that can be obtained by chitosan or chitin hydrolysis. Their enzymatic production is preferred over other methodologies (physical, chemical, etc.) due to the mild conditions required, the fewer amounts of waste and its efficiency to control product composition. By properly selecting the enzyme (chitinase, chitosanase or nonspecific enzymes) and the substrate properties (degree of deacetylation, molecular weight, etc.), it is possible to direct the synthesis towards any of the three COS types: fully acetylated (faCOS), partially acetylated (paCOS) and fully deacetylated (fdCOS). In this article, we review the main strategies to steer the COS production towards a specific group. The chemical characterization of COS by advanced techniques, e.g., high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and MALDI-TOF mass spectrometry, is critical for structure–function studies. The scaling of processes to synthesize specific COS mixtures is difficult due to the low solubility of chitin/chitosan, the heterogeneity of the reaction mixtures, and high amounts of salts. Enzyme immobilization can help to minimize such hurdles. The main bioactive properties of COS are herein reviewed. Finally, the anti-inflammatory activity of three COS mixtures was assayed in murine macrophages after stimulation with lipopolysaccharides.

scholarly journals A New Breeding Strategy towards Introgression and Characterization of Stay-Green QTL for Drought Tolerance in Sorghum

Agriculture ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 598
Author(s):  
Nasrein Mohamed Kamal ◽  
Yasir Serag Alnor Gorafi ◽  
Hanan Abdeltwab ◽  
Ishtiag Abdalla ◽  
Hisashi Tsujimoto ◽  
...  
Keyword(s):  
Phenotypic Expression ◽  
Phenotypic Selection ◽  
Breeding Strategy ◽  
Stay Green ◽  
Breeding Programs ◽  
The Public ◽  
Drought Tolerant ◽  
Selection Step ◽  
The Cost

Several marker-assisted selection (MAS) or backcrossing (MAB) approaches exist for polygenic trait improvement. However, the implementation of MAB remains a challenge in many breeding programs, especially in the public sector. In MAB introgression programs, which usually do not include phenotypic selection, undesired donor traits may unexpectedly turn up regardless of how expensive and theoretically powerful a backcross scheme may be. Therefore, combining genotyping and phenotyping during selection will improve understanding of QTL interactions with the environment, especially for minor alleles that maximize the phenotypic expression of the traits. Here, we describe the introgression of stay-green QTL (Stg1–Stg4) from B35 into two sorghum backgrounds through an MAB that combines genotypic and phenotypic (C-MAB) selection during early backcross cycles. The background selection step is excluded. Since it is necessary to decrease further the cost associated with molecular marker assays, the costs of C-MAB were estimated. Lines with stay-green trait and good performance were identified at an early backcross generation, backcross two (BC2). Developed BC2F4 lines were evaluated under irrigated and drought as well as three rainfed environments varied in drought timing and severity. Under drought conditions, the mean grain yield of the most C-MAB-introgression lines was consistently higher than that of the recurrent parents. This study is one of the real applications of the successful use of C-MAB for the development of drought-tolerant sorghum lines for drought-prone areas.

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