scholarly journals Lyophilization and homogenization of biological samples improves reproducibility and reduces standard deviation in molecular biology techniques

Amino Acids ◽  
2021 ◽  
Author(s):  
Agnes Molnar ◽  
Tamas Lakat ◽  
Adam Hosszu ◽  
Beata Szebeni ◽  
Anna Balogh ◽  
...  
Keyword(s):  
Biological Samples ◽  
Basic Research ◽  
Cost Effective ◽  
Specific Reference ◽  
Metabolome Analysis ◽  
C Storage ◽  
Mouse Tissues ◽  
Freeze Dried ◽  
Wide Range ◽  
Similar Quantity

AbstractLyophilization is a cost-effective method for biological specimen preservation but detailed tissue-specific reference protocols are still lacking. Moreover, data are limited on the long-term stability of proteins and nucleic acids in lyophilized samples.Here, we offer lyophilization protocols for various rat and mouse tissues (kidney, heart, liver, lung, aorta, and skin) coupled with technical hints for optimal sample preparation. We demonstrate that lyophilized samples stored at 4 °C for 20 months can yield protein and RNA of similar quantity and quality to −80 °C storage, while phosphorylated proteins are preserved as well. Freeze-dried and subsequently pulverized samples can provide more consistent, more reliable data especially when investigating focal injuries, such as fibrosis. We developed a protocol for the concentration of biological solutions and achieved 20-times concentration in human peritoneal dialysis effluent solution which enables the previously unattainable detection of proteins in these samples. We established a method for water removal as well as accurate water content measurement of fecal samples, which can be valuable for gut metabolome analysis.Taken together, lyophilization is a valuable tool for the preservation of biological samples with many advantages. We aim to draw attention to the wide range of possibilities offered by freeze drying in pre-clinical or basic research.

A Disposable Electrochemical Sensor Based on Iron Molybdate for Analysis of Dopamine in Biological Samples

2021 ◽  
Author(s):  
E Ashok Kumar ◽  
Tse-Wei Chen ◽  
Shen-Ming Chen ◽  
Tzyy-Jiann Wang ◽  
Allen J. A ◽  
...  
Keyword(s):  
Electrochemical Sensor ◽  
Biological Samples ◽  
Basic Research ◽  
Cost Effective ◽  
Electrochemical Sensing ◽  
Iron Molybdate ◽  
Sensing Platform ◽  
Electrochemical Devices ◽  
Care Technology

Developing cost-effective approaches for the fabrication of electrochemical devices is instantly needed for transferring from basic research to point-care technology. Herein, we report an electrochemical sensing platform based on iron...

scholarly journals Rapid and scalable preparation of bacterial lysates for cell-free gene expression

2017 ◽  
Author(s):  
Andriy Didovyk ◽  
Taishi Tonooka ◽  
Lev Tsimring ◽  
Jeff Hasty
Keyword(s):  
Gene Expression ◽  
Synthetic Biology ◽  
Cost Effective ◽  
Diverse Range ◽  
Highly Active ◽  
Freeze Dried ◽  
Wide Range ◽  
Specialized Equipment ◽  
Free Gene ◽  
Bacterial Lysates

AbstractCell-free gene expression systems are emerging as an important platform for a diverse range of synthetic biology and biotechnology applications, including production of robust field-ready biosensors. Here, we combine programmed cellular autolysis with a freeze-thaw or freeze-dry cycle to create a practical, reproducible, and a labor- and cost-effective approach for rapid production of bacterial lysates for cell-free gene expression. Using this method, ro-bust and highly active bacterial cell lysates can be produced without specialized equipment at a wide range of scales, making cell-free gene expression easily and broadly accessible. More-over, live autolysis strain can be freeze-dried directly and subsequently lysed upon rehydration to produce active lysate. We demonstrate the utility of autolysates for synthetic biology by reg-ulating protein production and degradation, implementing quorum sensing, and showing quan-titative protection of linear DNA templates by GamS protein. To allow versatile and sensitive β-galactosidase (LacZ) based readout we produce autolysates with no detectable background LacZ activity and use them to produce sensitive mercury(II) biosensors with LacZ-mediated colorimetric and fluorescent outputs. The autolysis approach can facilitate wider adoption of cell-free technology for cell-free gene expression as well as other synthetic biology and biotechnology applications, such as metabolic engineering, natural product biosynthesis, or proteomics.

CLEX: A cross-linguistic lexical norms database*

2009 ◽  
Vol 37 (2) ◽  
pp. 419-428 ◽  
Author(s):  
RUNE NØRGAARD JØRGENSEN ◽  
PHILIP S. DALE ◽  
DORTHE BLESES ◽  
LARRY FENSON
Keyword(s):  
Effective Means ◽  
Basic Research ◽  
Cost Effective ◽  
Parent Report ◽  
Research Assessment ◽  
Lexical Development ◽  
Communicative Development ◽  
Lexical Norms ◽  
Wide Range ◽  
Lexical Data

ABSTRACTParent report has proven a valid and cost-effective means of evaluating early child language. Norming datasets for these instruments, which provide the basis for standardized comparisons of individual children to a population, can also be used to derive norms for the acquisition of individual words in production and comprehension and also early gestures and symbolic actions. These lexical norms have a wide range of uses in basic research, assessment and intervention. In addition, cross-linguistic comparisons of lexical development are greatly facilitated by the availability of norms from diverse languages. This report describes the development of CLEX, a new web-based cross-linguistic database for lexical data from adaptations of the MacArthur-Bates Communicative Development Inventories. CLEX provides tools for a range of analyses within and across languages. It is designed to incorporate additional language datasets easily, and to permit users to define mappings between lexical items in pairs of languages for more specific cross-linguistic comparisons.

Artefacts in the x-ray microanalysis of frozen-hydrated biological samples

1987 ◽  
Vol 45 ◽  
pp. 658-659
Author(s):  
Patrick Echlin
Keyword(s):  
Electron Scattering ◽  
Biological Samples ◽  
Fracture Face ◽  
Detailed Structure ◽  
X Ray ◽  
Morphological Appearance ◽  
The Poor ◽  
Freeze Dried

A number of papers have appeared recently which purport to have carried out x-ray microanalysis on fully frozen hydrated samples. It is important to establish reliable criteria to be certain that a sample is in a fully hydrated state. The morphological appearance of the sample is an obvious parameter because fully hydrated samples lack the detailed structure seen in their freeze dried counterparts. The electron scattering by ice within a frozen-hydrated section and from the surface of a frozen-hydrated fracture face obscures cellular detail. (Fig. 1G and 1H.) However, the morphological appearance alone can be quite deceptive for as Figures 1E and 1F show, parts of frozen-dried samples may also have the poor morphology normally associated with fully hydrated samples. It is only when one examines the x-ray spectra that an assurance can be given that the sample is fully hydrated.

2D Color-Intensity Representation of Ultrasonic Thickness Gauge Measurements

2020 ◽  
pp. 1192-1198
Author(s):  
M.S. Mohammad ◽  
Tibebe Tesfaye ◽  
Kim Ki-Seong
Keyword(s):  
Cost Effective ◽  
The Other ◽  
Thickness Gauge ◽  
Color Intensity ◽  
Digital Readout ◽  
Flat Surfaces ◽  
Video Cameras ◽  
Wide Range ◽  
Cost Effective Alternative ◽  
Ultrasonic Thickness

Ultrasonic thickness gauges are easy to operate and reliable, and can be used to measure a wide range of thicknesses and inspect all engineering materials. Supplementing the simple ultrasonic thickness gauges that present results in either a digital readout or as an A-scan with systems that enable correlating the measured values to their positions on the inspected surface to produce a two-dimensional (2D) thickness representation can extend their benefits and provide a cost-effective alternative to expensive advanced C-scan machines. In previous work, the authors introduced a system for the positioning and mapping of the values measured by the ultrasonic thickness gauges and flaw detectors (Tesfaye et al. 2019). The system is an alternative to the systems that use mechanical scanners, encoders, and sophisticated UT machines. It used a camera to record the probe’s movement and a projected laser grid obtained by a laser pattern generator to locate the probe on the inspected surface. In this paper, a novel system is proposed to be applied to flat surfaces, in addition to overcoming the other limitations posed due to the use of the laser projection. The proposed system uses two video cameras, one to monitor the probe’s movement on the inspected surface and the other to capture the corresponding digital readout of the thickness gauge. The acquired images of the probe’s position and thickness gauge readout are processed to plot the measured data in a 2D color-coded map. The system is meant to be simpler and more effective than the previous development.

The Future of Plasma-Based Surface Engineering Techniques in Tribology (Keynote)

2005 ◽  
Author(s):  
Allan Matthews ◽  
Adrian Leyland
Keyword(s):  
Surface Engineering ◽  
Performance Enhancement ◽  
Bulk Material ◽  
Cost Savings ◽  
Cost Effective ◽  
Ceramic Coatings ◽  
Performance Improvements ◽  
Treatment Techniques ◽  
Wide Range ◽  
Macro Scale

Over the past twenty years or so, there have been major steps forward both in the understanding of tribological mechanisms and in the development of new coating and treatment techniques to better “engineer” surfaces to achieve reductions in wear and friction. Particularly in the coatings tribology field, improved techniques and theories which enable us to study and understand the mechanisms occurring at the “nano”, “micro” and “macro” scale have allowed considerable progress to be made in (for example) understanding contact mechanisms and the influence of “third bodies” [1–5]. Over the same period, we have seen the emergence of the discipline which we now call “Surface Engineering”, by which, ideally, a bulk material (the ‘substrate’) and a coating are combined in a way that provides a cost-effective performance enhancement of which neither would be capable without the presence of the other. It is probably fair to say that the emergence and recognition of Surface Engineering as a field in its own right has been driven largely by the availability of “plasma”-based coating and treatment processes, which can provide surface properties which were previously unachievable. In particular, plasma-assisted (PA) physical vapour deposition (PVD) techniques, allowing wear-resistant ceramic thin films such as titanium nitride (TiN) to be deposited on a wide range of industrial tooling, gave a step-change in industrial productivity and manufactured product quality, and caught the attention of engineers due to the remarkable cost savings and performance improvements obtained. Subsequently, so-called 2nd- and 3rd-generation ceramic coatings (with multilayered or nanocomposite structures) have recently been developed [6–9], to further extend tool performance — the objective typically being to increase coating hardness further, or extend hardness capabilities to higher temperatures.

scholarly journals Gamma models for estimating the odds ratio for a skewed biomarker measured in pools and subject to errors

Biostatistics ◽  
2019 ◽  
Author(s):  
Dane R Van Domelen ◽  
Emily M Mitchell ◽  
Neil J Perkins ◽  
Enrique F Schisterman ◽  
Amita K Manatunga ◽  
...  
Keyword(s):  
Odds Ratio ◽  
Adjusted Odds Ratio ◽  
Lower Cost ◽  
Cost Effective ◽  
Ratio Estimation ◽  
Statistical Efficiency ◽  
Function Model ◽  
Processing Error ◽  
Wide Range ◽  
Pooled Samples

SUMMARYMeasuring a biomarker in pooled samples from multiple cases or controls can lead to cost-effective estimation of a covariate-adjusted odds ratio, particularly for expensive assays. But pooled measurements may be affected by assay-related measurement error (ME) and/or pooling-related processing error (PE), which can induce bias if ignored. Building on recently developed methods for a normal biomarker subject to additive errors, we present two related estimators for a right-skewed biomarker subject to multiplicative errors: one based on logistic regression and the other based on a Gamma discriminant function model. Applied to a reproductive health dataset with a right-skewed cytokine measured in pools of size 1 and 2, both methods suggest no association with spontaneous abortion. The fitted models indicate little ME but fairly severe PE, the latter of which is much too large to ignore. Simulations mimicking these data with a non-unity odds ratio confirm validity of the estimators and illustrate how PE can detract from pooling-related gains in statistical efficiency. These methods address a key issue associated with the homogeneous pools study design and should facilitate valid odds ratio estimation at a lower cost in a wide range of scenarios.

EXPRESS: Method dependent variation in TSH and FT4 reference intervals in pregnancy: a systematic review

2021 ◽  
pp. 000456322110269
Author(s):  
O E Okosieme ◽  
Medha Agrawal ◽  
Danyal Usman ◽  
Carol Evans
Keyword(s):  
Systematic Review ◽  
Reference Interval ◽  
First Trimester ◽  
Reference Intervals ◽  
Assay Method ◽  
Specific Reference ◽  
Upper Limits ◽  
Wide Range ◽  
Assay Methods ◽  
In Pregnancy

Background: Gestational TSH and FT4 reference intervals may differ according to assay method but the extent of variation is unclear and has not been systematically evaluated. We conducted a systematic review of published studies on TSH and FT4 reference intervals in pregnancy. Our aim was to quantify method-related differences in gestation reference intervals, across four commonly used assay methods, Abbott, Beckman, Roche, and Siemens. Methods: We searched the literature for relevant studies, published between January 2000 and December 2020, in healthy pregnant women without thyroid antibodies or disease. For each study, we extracted trimester-specific reference intervals (2.5–97.5 percentiles) for TSH and FT4 as well as the manufacturer provided reference interval for the corresponding non-pregnant population. Results: TSH reference intervals showed a wide range of study-to-study differences with upper limits ranging from 2.33 to 8.30 mU/L. FT4 lower limits ranged from 4.40–13.93 pmol/L, with consistently lower reference intervals observed with the Beckman method. Differences between non-pregnant and first trimester reference intervals were highly variable, and for most studies the TSH upper limit in the first trimester could not be predicted or extrapolated from non-pregnant values. Conclusions: Our study confirms significant intra and inter-method disparities in gestational thyroid hormone reference intervals. The relationship between pregnant and non-pregnant values is inconsistent and does not support the existing practice in some laboratories of extrapolating gestation references from non-pregnant values. Laboratories should invest in deriving method-specific gestation reference intervals for their population.

scholarly journals Sporulation in solventogenic and acetogenic clostridia

2021 ◽  
Author(s):  
Mamou Diallo ◽  
Servé W. M. Kengen ◽  
Ana M. López-Contreras
Keyword(s):  
Current Knowledge ◽  
Carbon Sources ◽  
Cost Effective ◽  
Biotechnological Production ◽  
Key Points ◽  
Wide Range ◽  
Potential Applications ◽  
A Cell ◽  
Sporulation Initiation ◽  
Solventogenic Clostridia

AbstractThe Clostridium genus harbors compelling organisms for biotechnological production processes; while acetogenic clostridia can fix C1-compounds to produce acetate and ethanol, solventogenic clostridia can utilize a wide range of carbon sources to produce commercially valuable carboxylic acids, alcohols, and ketones by fermentation. Despite their potential, the conversion by these bacteria of carbohydrates or C1 compounds to alcohols is not cost-effective enough to result in economically viable processes. Engineering solventogenic clostridia by impairing sporulation is one of the investigated approaches to improve solvent productivity. Sporulation is a cell differentiation process triggered in bacteria in response to exposure to environmental stressors. The generated spores are metabolically inactive but resistant to harsh conditions (UV, chemicals, heat, oxygen). In Firmicutes, sporulation has been mainly studied in bacilli and pathogenic clostridia, and our knowledge of sporulation in solvent-producing or acetogenic clostridia is limited. Still, sporulation is an integral part of the cellular physiology of clostridia; thus, understanding the regulation of sporulation and its connection to solvent production may give clues to improve the performance of solventogenic clostridia. This review aims to provide an overview of the triggers, characteristics, and regulatory mechanism of sporulation in solventogenic clostridia. Those are further compared to the current knowledge on sporulation in the industrially relevant acetogenic clostridia. Finally, the potential applications of spores for process improvement are discussed.Key Points• The regulatory network governing sporulation initiation varies in solventogenic clostridia.• Media composition and cell density are the main triggers of sporulation.• Spores can be used to improve the fermentation process.

scholarly journals Role of Insulin in Health and Disease: An Update

2021 ◽  
Vol 22 (12) ◽  
pp. 6403
Author(s):  
Md Saidur Rahman ◽  
Khandkar Shaharina Hossain ◽  
Sharnali Das ◽  
Sushmita Kundu ◽  
Elikanah Olusayo Adegoke ◽  
...  
Keyword(s):  
Insulin Signaling ◽  
Basic Research ◽  
Future Research ◽  
Protective Effects ◽  
Glucose Levels ◽  
Physiological Processes ◽  
Blood Glucose Levels ◽  
Wide Range ◽  
Health And Disease

Insulin is a polypeptide hormone mainly secreted by β cells in the islets of Langerhans of the pancreas. The hormone potentially coordinates with glucagon to modulate blood glucose levels; insulin acts via an anabolic pathway, while glucagon performs catabolic functions. Insulin regulates glucose levels in the bloodstream and induces glucose storage in the liver, muscles, and adipose tissue, resulting in overall weight gain. The modulation of a wide range of physiological processes by insulin makes its synthesis and levels critical in the onset and progression of several chronic diseases. Although clinical and basic research has made significant progress in understanding the role of insulin in several pathophysiological processes, many aspects of these functions have yet to be elucidated. This review provides an update on insulin secretion and regulation, and its physiological roles and functions in different organs and cells, and implications to overall health. We cast light on recent advances in insulin-signaling targeted therapies, the protective effects of insulin signaling activators against disease, and recommendations and directions for future research.

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