scholarly journals Decentralizing Cell-Free RNA Sensing With the Use of Low-Cost Cell Extracts

2021 ◽  
Vol 9 ◽  
Author(s):  
Anibal Arce ◽  
Fernando Guzman Chavez ◽  
Chiara Gandini ◽  
Juan Puig ◽  
Tamara Matute ◽  
...  
Keyword(s):  
Low Cost ◽  
Cold Chain ◽  
Expression Systems ◽  
Cell Extracts ◽  
Freeze Dried ◽  
Rna Sensors ◽  
Pure Cell ◽  
Pcr Products ◽  
Free Gene ◽  
The Stability

Cell-free gene expression systems have emerged as a promising platform for field-deployed biosensing and diagnostics. When combined with programmable toehold switch-based RNA sensors, these systems can be used to detect arbitrary RNAs and freeze-dried for room temperature transport to the point-of-need. These sensors, however, have been mainly implemented using reconstituted PURE cell-free protein expression systems that are difficult to source in the Global South due to their high commercial cost and cold-chain shipping requirements. Based on preliminary demonstrations of toehold sensors working on lysates, we describe the fast prototyping of RNA toehold switch-based sensors that can be produced locally and reduce the cost of sensors by two orders of magnitude. We demonstrate that these in-house cell lysates provide sensor performance comparable to commercial PURE cell-free systems. We further optimize these lysates with a CRISPRi strategy to enhance the stability of linear DNAs by knocking-down genes responsible for linear DNA degradation. This enables the direct use of PCR products for fast screening of new designs. As a proof-of-concept, we develop novel toehold sensors for the plant pathogen Potato Virus Y (PVY), which dramatically reduces the yield of this important staple crop. The local implementation of low-cost cell-free toehold sensors could enable biosensing capacity at the regional level and lead to more decentralized models for global surveillance of infectious disease.

scholarly journals Decentralizing cell-free RNA sensing with the use of low-cost cell extracts

2021 ◽  
Author(s):  
Anibal Arce ◽  
Fernando Guzman ◽  
Chiara Gandini ◽  
Juan Puig ◽  
Tamara Matute ◽  
...  
Keyword(s):  
Protein Expression ◽  
Low Cost ◽  
Cold Chain ◽  
Expression Systems ◽  
Free Protein ◽  
Cell Extracts ◽  
Freeze Dried ◽  
Rna Sensors ◽  
Pure Cell ◽  
Pcr Products

Cell-free gene expression systems have emerged as a promising platform for field-deployed biosensing and diagnostics. When combined with programmable toehold switch-based RNA sensors, these systems can be used to detect arbitrary RNAs and freeze-dried for room temperature transport to the point-of-need. These sensors, however, have been implemented using reconstituted PURE cell-free protein expression systems that are difficult to source in the Global South due to their high commercial cost and cold-chain shipping requirements. Here, we describe the implementation of RNA toehold switch-based sensors using E. coli cell lysate-based cell-free protein expression systems, which can be produced locally and reduce the cost of sensors by two orders of magnitude. We then demonstrate that these in-house cell lysates provide sensor performance comparable to commercial PURE cell-free systems. We further optimize use of these lysates with a CRISPRi strategy to enhance the stability of linear DNAs, enabling the direct use of PCR products for fast screening of new designs. As a proof-of-concept, we develop novel toeholds sensors for the plant pathogen Potato Virus Y (PVY), which dramatically reduces the yield of this important staple crop. The local implementation of low-cost cell-free toehold sensors could enable biosensing capacity at the regional level and lead to more decentralized models for global surveillance of infectious disease.

The Stability of the WHO Reference Thromboplastin NIBS & C 67/40

1979 ◽  
Vol 42 (04) ◽  
pp. 1135-1140 ◽  
Author(s):  
G I C Ingram
Keyword(s):  
Human Brain ◽  
Collaborative Study ◽  
Calibration Constant ◽  
Long Term Stability ◽  
Freeze Dried ◽  
Show Evidence ◽  
Human Material ◽  
Reference Preparation ◽  
The Stability

SummaryThe International Reference Preparation of human brain thromboplastin coded 67/40 has been thought to show evidence of instability. The evidence is discussed and is not thought to be strong; but it is suggested that it would be wise to replace 67/40 with a new preparation of human brain, both for this reason and because 67/40 is in a form (like Thrombotest) in which few workers seem to use human brain. A �plain� preparation would be more appropriate; and a freeze-dried sample of BCT is recommended as the successor preparation. The opportunity should be taken also to replace the corresponding ox and rabbit preparations. In the collaborative study which would be required it would then be desirable to test in parallel the three old and the three new preparations. The relative sensitivities of the old preparations could be compared with those found in earlier studies to obtain further evidence on the stability of 67/40; if stability were confirmed, the new preparations should be calibrated against it, but if not, the new human material should receive a calibration constant of 1.0 and the new ox and rabbit materials calibrated against that.The types of evidence available for monitoring the long-term stability of a thromboplastin are discussed.

scholarly journals Exploring the Structural Competition between the Black and the Yellow Phase of CsPbI3

Nanomaterials ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1282
Author(s):  
Ioannis Deretzis ◽  
Corrado Bongiorno ◽  
Giovanni Mannino ◽  
Emanuele Smecca ◽  
Salvatore Sanzaro ◽  
...  
Keyword(s):  
Perovskite Phase ◽  
Low Cost ◽  
Δ Phase ◽  
Nanocrystalline Structures ◽  
Halide Perovskites ◽  
The Stability ◽  
Structural Competition ◽  
Lead Halide ◽  
Main Candidate ◽  
Spontaneous Phase

The realization of stable inorganic perovskites is crucial to enable low-cost solution-processed photovoltaics. However, the main candidate material, CsPbI3, suffers from a spontaneous phase transition at room temperature towards a photo-inactive orthorhombic δ-phase (yellow phase). Here we used theoretical and experimental methods to study the structural and electronic features that determine the stability of the CsPbI3 perovskite. We argued that the two physical characteristics that favor the black perovskite phase at low temperatures are the strong spatial confinement in nanocrystalline structures and the level of electron doping in the material. Within this context, we discussed practical procedures for the realization of long-lasting inorganic lead halide perovskites.

scholarly journals Longevity of Raw and Lyophilized Crude Urease Extracts

2021 ◽  
Vol 2 (2) ◽  
pp. 325-334
Author(s):  
Neda Javadi ◽  
Hamed Khodadadi Tirkolaei ◽  
Nasser Hamdan ◽  
Edward Kavazanjian
Keyword(s):  
Crude Extract ◽  
Room Temperature ◽  
Low Cost ◽  
Extraction Process ◽  
Crude Extracts ◽  
Simple Extraction ◽  
Commercial Applications ◽  
Stable Source ◽  
The Stability ◽  
The Cost

The stability (longevity of activity) of three crude urease extracts was evaluated in a laboratory study as part of an effort to reduce the cost of urease for applications that do not require high purity enzyme. A low-cost, stable source of urease will greatly facilitate engineering applications of urease such as biocementation of soil. Inexpensive crude extracts of urease have been shown to be effective at hydrolyzing urea for carbonate precipitation. However, some studies have suggested that the activity of a crude extract may decrease with time, limiting the potential for its mass production for commercial applications. The stability of crude urease extracts shown to be effective for biocementation was studied. The crude extracts were obtained from jack beans via a simple extraction process, stored at room temperature and at 4 ℃, and periodically tested to evaluate their stability. To facilitate storage and transportation of the extracted enzyme, the longevity of the enzyme following freeze drying (lyophilization) to reduce the crude extract to a powder and subsequent re-hydration into an aqueous solution was evaluated. In an attempt to improve the shelf life of the lyophilized extract, dextran and sucrose were added during lyophilization. The stability of purified commercial urease following rehydration was also investigated. Results of the laboratory tests showed that the lyophilized crude extract maintained its activity during storage more effectively than either the crude extract solution or the rehydrated commercial urease. While incorporating 2% dextran (w/v) prior to lyophilization of the crude extract increased the overall enzymatic activity, it did not enhance the stability of the urease during storage.

scholarly journals A Real-Time Method for Improving Stability of Monolithic Quartz Crystal Microbalance Operating under Harsh Environmental Conditions

Sensors ◽  
2021 ◽  
Vol 21 (12) ◽  
pp. 4166
Author(s):  
Román Fernández ◽  
María Calero ◽  
Yolanda Jiménez ◽  
Antonio Arnau
Keyword(s):  
Real Time ◽  
Quartz Crystal Microbalance ◽  
Quartz Crystal ◽  
Limit Of Detection ◽  
Quartz Substrate ◽  
Low Cost ◽  
Discrete Wavelet ◽  
Reagent Consumption ◽  
Measurement Cell ◽  
The Stability

Monolithic quartz crystal microbalance (MQCM) has recently emerged as a very promising technology suitable for biosensing applications. These devices consist of an array of miniaturized QCM sensors integrated within the same quartz substrate capable of detecting multiple target analytes simultaneously. Their relevant benefits include high throughput, low cost per sensor unit, low sample/reagent consumption and fast sensing response. Despite the great potential of MQCM, unwanted environmental factors (e.g., temperature, humidity, vibrations, or pressure) and perturbations intrinsic to the sensor setup (e.g., mechanical stress exerted by the measurement cell or electronic noise of the characterization system) can affect sensor stability, masking the signal of interest and degrading the limit of detection (LoD). Here, we present a method based on the discrete wavelet transform (DWT) to improve the stability of the resonance frequency and dissipation signals in real time. The method takes advantage of the similarity among the noise patterns of the resonators integrated in an MQCM device to mitigate disturbing factors that impact on sensor response. Performance of the method is validated by studying the adsorption of proteins (neutravidin and biotinylated albumin) under external controlled factors (temperature and pressure/flow rate) that simulate unwanted disturbances.

Modal Analyses of Deployable Truss Structures Based on Shape Memory Polymer Composites

2016 ◽  
Vol 08 (07) ◽  
pp. 1640009 ◽  
Author(s):  
Fengfeng Li ◽  
Liwu Liu ◽  
Xin Lan ◽  
Tong Wang ◽  
Xiangyu Li ◽  
...  
Keyword(s):  
Finite Element ◽  
Shape Memory ◽  
Polymer Composites ◽  
Dynamic Properties ◽  
Low Cost ◽  
Shape Memory Polymer ◽  
Truss Structures ◽  
Modal Test ◽  
The Stability ◽  
Mechanical Devices

With large spatial deployable antennas used more widely, the stability of deployable antennas is attracting more attention. The form of the support structure is an important factor of the antenna’s natural frequency, which is essential to study to prevent the resonance. The deployable truss structures based on shape memory polymer composites (SMPCs) have made themselves feasible for their unique properties such as highly reliable, low-cost, light weight, and self-deployment without complex mechanical devices compared with conventional deployable masts. This study offers deliverables as follows: an establishment of three-longeron beam and three-longeron truss finite element models by using ABAQUS; calculation of natural frequencies and vibration modes; parameter studies for influence on their dynamic properties; manufacture of a three-longeron truss based on SMPC, and modal test of the three-longeron truss. The results show that modal test and finite element simulation fit well.

Thinking about Jointly Preventing the “Color Revolution” by China and Russia

2021 ◽  
pp. 100
Author(s):  
chensheng wang
Keyword(s):  
International System ◽  
Low Cost ◽  
The United States ◽  
Regional Security ◽  
Healthy Development ◽  
Regional Coordination ◽  
Color Revolution ◽  
Return Method ◽  
The Stability ◽  
Surrounding Areas

The color revolution, which is a “low-cost and high-return” method in regime change, has become the main mean and priority option for America to subvert dissident regimes. In recent years, with the raising strength of containing and suppressing between China with Russia by the United States, America has tried its best to plan “color revolution” not only around China and Russia, but also within the borders of the two countries. China and Russia have become the key target of America in implementing the “color revolution”, however, the situation of the two countries to prevent the “color revolution” is particularly urgent. The “color revolution” not only disrupts the balance of the international system and regional security, but also seriously affects the stability of the country's political power and the healthy development of the economy. In view of this, it is now necessary for China and Russia to work together to prevent “color revolution”. Regarding the new changes, methods changed from non-violent to violent me, more advanced organizational methods, the younger generation of the participants, and changes in manifestations by the “color revolution”, as well as the underlying causes of the “color revolution”, China and Russia should have uindividualized strategies. China and Russia can strengthen cooperation in different areas, such as politics, economy, culture, ideological education, and regional coordination. China and Russia should take advantages of their respective experiences in dealing with “color revolution”, strengthen sharing and communicating experience with other countries in the region, and jointly build a barrier to prevent “color revolution” and protect the security and stability in China and Russia and the surrounding areas.

672 An Investigation of Antibiotic Loading and Release in Acid-precipitated Keratin Proteins to Treat Topical Burn Infections

2021 ◽  
Vol 42 (Supplement_1) ◽  
pp. S192-S193
Author(s):  
Marc A Thompson ◽  
Robert J Christy
Keyword(s):  
Proteolytic Enzymes ◽  
Diglycidyl Ether ◽  
Water Soluble ◽  
Bacterial Colony ◽  
Freeze Dried ◽  
Antibiotic Compounds ◽  
Water Ph ◽  
Stable Product ◽  
Antibiotic Release ◽  
The Stability

Abstract Introduction Human-hair derived keratin (KOS) protein has been selected in this investigation for its ability to bind antibiotic compounds and provide sustained release while withstanding harsh proteolytic environments such as inflamed, damaged tissue. The need to control local flora has been recognized as an imperative for wound healing, as recovery is significantly hampered by infection. This study investigates the synthesis of KOS-based particulate matter, developed using acid-precipitation, to load and release the water-soluble antibiotic ciprofloxacin (CIP). We hypothesize that ionically bound CIP release is tied to the degradation of KOS, therefore, bacterial metabolism, which produces proteolytic enzymes, will trigger CIP release thereby creating a novel self-extinguishing delivery system for contaminated skin wounds. Methods Ciprofloxacin hydrochloride was solubilized in deionized water (pH 5.3) under constant stirring. Freeze-dried KOS powder was added for an ultimately 5% w/v and 0.8% w/v solution of KOS and CIP, respectively. To improve the stability of KOS a water-soluble diglycidyl ether crosslinker was added to solutions and stirred for 24 hours. CIP-loaded protein was precipitated out by a hydrochloric acid induced pH reduction. Samples were collected and frozen at -20 °C prior to lyophilization, thus forming the stable product. Degradation of KOS and commensurate release of CIP were measured using a bicinchoninic acid (BCA) assay and fluorescent measurements of hydrated material supernatant. The reduction of bacterial colonies was validated by a broth inhibition assay whereby CIP-loaded KOS or unloaded KOS controls where hydrated in bacterial-laden broth cultures of Pseudomonas aeruginosa or Methicillin-resistant Staphylococcus aureus. Cultures were sampled at 24, 48, or 72 hours and plated to quantify colony-forming units. Results The presence of CIP in the KOS protein was confirmed and release rates follow similar patterns to that of KOS degradation. CIP-loaded proteins significantly reduce bacterial colony presence in concentrated inoculant solutions up to 72 hours. Conclusions CIP release does appear to coincide with KOS degradation, which is bolstered in the presence of infectious levels of bacteria. Ongoing studies aim to observe more robust models of infection and more controlled antibiotic release.

scholarly journals An Accurate DDS Method Using Compound Frequency Tuning Word and Its FPGA Implementation

Electronics ◽  
2018 ◽  
Vol 7 (11) ◽  
pp. 330 ◽  
Author(s):  
Yuqing Hou ◽  
Changlong Li ◽  
Sheng Tang
Keyword(s):  
Frequency Synthesizer ◽  
Frequency Tuning ◽  
Low Cost ◽  
Atomic Clock ◽  
Frequency Error ◽  
Frequency Synthesis ◽  
Direct Digital Synthesizer ◽  
Low Power Dissipation ◽  
Conversion Time ◽  
The Stability

Because of its high resolution, low cost, small volume, low power dissipation and less conversion time consumption, the direct digital synthesizer (DDS) method has been applied more and more in the fields of frequency synthesis and signal generation. However, only a limited number of precise frequency signals can be synthesized by the traditional DDS, for the reason that its accumulator modulus is fixed, and its frequency tuning word must be integer. In this paper, a precise DDS method using compound frequency tuning word is proposed, which improves the accuracy of synthesized signals at any frequency points on the premise of guaranteeing the stability of synthesized signals. In order to verify the effectiveness of the new method, a DDS frequency synthesizer based on FPGA is designed and implemented. Taking the rubidium atomic clock PRS10 as standard frequency source, the experiments shows that the frequency stability of the synthesized signal is better than 8.0 × 10−12/s, the relative frequency error is less than 4.8 × 10−12, and that the frequency accuracy is improved by three orders of magnitude compared with the traditional DDS method.

Sign in / Sign up

Export Citation Format

Share Document