Design of experiments Dotaz Zobrazit nápovědu
Nanosized drug crystals have been reported with enhanced apparent solubility, bioavailability, and therapeutic efficacy compared to microcrystal materials, which are not suitable for parenteral administration. However, nanocrystal design and development by bottom-up approaches are challenging, especially considering the non-standardized process parameters in the injection step. This work aims to present a systematic step-by-step approach through Quality-by-Design (QbD) and Design of Experiments (DoE) for synthesizing drug nanocrystals by a semi-automated nanoprecipitation method. Curcumin is used as a drug model due to its well-known poor water solubility (0.6 µg mL-1, 25 °C). Formal and informal risk assessment tools allow identifying the critical factors. A fractional factorial 24-1 screening design evaluates their impact on the average size and polydispersity of nanocrystals. The optimization of significant factors is done by a Central Composite Design. This response surface methodology supports the rational design of the nanocrystals, identifying and exploring the design space. The proposed joint approach leads to a reproducible, robust, and stable nanocrystalline preparation of 316 nm with a PdI of 0.217 in compliance with the quality profile. An orthogonal approach for particle size and polydispersity characterization allows discarding the formation of aggregates. Overall, the synergy between advanced data analysis and semi-automated standardized nanocrystallization of drugs is highlighted.
- Klíčová slova
- design space, nanocrystals, orthogonal characterization, response surface methodology, solvent–antisolvent precipitation,
- MeSH
- automatizace MeSH
- krystalizace MeSH
- kurkumin chemie MeSH
- léčivé přípravky chemie MeSH
- nanočástice * chemie MeSH
- velikost částic MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- kurkumin MeSH
- léčivé přípravky MeSH
Dual reporters encoding two distinct proteins within the same mRNA have had a crucial role in identifying and characterizing unconventional mechanisms of eukaryotic translation. These mechanisms include initiation via internal ribosomal entry sites (IRESs), ribosomal frameshifting, stop codon readthrough and reinitiation. This design enables the expression of one reporter to be influenced by the specific mechanism under investigation, while the other reporter serves as an internal control. However, challenges arise when intervening test sequences are placed between these two reporters. Such sequences can inadvertently impact the expression or function of either reporter, independent of translation-related changes, potentially biasing the results. These effects may occur due to cryptic regulatory elements inducing or affecting transcription initiation, splicing, polyadenylation and antisense transcription as well as unpredictable effects of the translated test sequences on the stability and activity of the reporters. Unfortunately, these unintended effects may lead to misinterpretation of data and the publication of incorrect conclusions in the scientific literature. To address this issue and to assist the scientific community in accurately interpreting dual-reporter experiments, we have developed comprehensive guidelines. These guidelines cover experimental design, interpretation and the minimal requirements for reporting results. They are designed to aid researchers conducting these experiments as well as reviewers, editors and other investigators who seek to evaluate published data.
- MeSH
- Eukaryota genetika MeSH
- lidé MeSH
- messenger RNA genetika metabolismus MeSH
- proteosyntéza genetika MeSH
- reportérové geny * MeSH
- směrnice jako téma MeSH
- výzkumný projekt normy MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- messenger RNA MeSH
Various adaptive designs have been proposed and applied to clinical trials, bioassay, psychophysics, etc. Adaptive designs are also useful in high cost engineering trials. More and more people have been paying attention to these design methods. This paper introduces several broad families of designs, such asthe play-the-winner rule, randomized play-the-winner rule and its generalization to the multi-arm case, doubly biased coin adaptive design, Markov chain model.
- MeSH
- klinické zkoušky jako téma klasifikace metody MeSH
- lidé MeSH
- Markovovy řetězce MeSH
- řízení kvality MeSH
- statistické modely * MeSH
- velikost vzorku MeSH
- výběr pacientů * MeSH
- výběrový bias MeSH
- výsledek terapie MeSH
- výzkumný projekt * MeSH
- zpětná vazba MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
To assess the environmental fate of engineered nanoparticles (ENPs), it is essential to understand their interactions with dissolved organic matter (DOM). The highly complex nature of the interactions between DOM and ENPs and other particulate matter (PM) requires investigating a wide range of material types under different conditions. However, despite repeated calls for an increased diversity of the DOM and PM studied, researchers increasingly focus on certain subsets of DOM and PM. Considering the discrepancy between the calls for more diversity and the research actually carried out, we hypothesize that materials that were studied more often are more visible in the scientific literature and therefore are more likely to be studied again. To investigate the plausibility of this hypothesis, we developed an agent-based model simulating the material choice in the experiments studying the interaction between DOM and PM between 1990 and 2015. The model reproduces the temporal trends in the choice of materials as well as the main properties of a network that displays the DOM and PM types investigated experimentally. The results, which support the hypothesis of a positive reinforcing material choice, help to explain why calls to increase the diversity of the materials studied are repeatedly made and why recent criticism states that the selection of materials is unbalanced.
- MeSH
- chemické látky znečišťující vodu analýza MeSH
- chemické modely MeSH
- huminové látky analýza MeSH
- monitorování životního prostředí metody MeSH
- nanočástice MeSH
- organické látky analýza MeSH
- pevné částice analýza MeSH
- počítačová simulace MeSH
- rozpustnost MeSH
- výběrový bias * MeSH
- výzkumný projekt MeSH
- zkreslení výsledků (epidemiologie) * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- chemické látky znečišťující vodu MeSH
- huminové látky MeSH
- organické látky MeSH
- pevné částice MeSH
This paper describes possible ways of prediction of nitrogen oxides formation during combustion of hydrocarbon fuels. Mathematical model based on experimental data acquired from the testing facility has been developed. The model enables to predict--at a high probability measure--the extent of nitrogen oxides emissions. The mathematical model of nitrogen oxide formation relies on the application of simplified kinetic equations describing the formation of nitrogen oxides at so-called equivalent temperature. It is a semi-empirical model that comes out of experimental knowledge. An important role played by the burner design itself has been emphasized and therefore an important supplementary parameter of the model is the characteristic of the burner design. It has been established that there was a good agreement between experimental data and those calculated by the application of the model to various conditions marked out by different combustion parameters in the combustion chamber. The results obtained by application of the model respect the influence of parameters validated by industrial practice that control the formation of nitrogen oxides in the course of fuel combustion. Such parameters-first of all-tare the temperature in the combustion chamber and the concentration of the substances taking part in the reaction. By application of the model, it is possible to assess the consequence of, for example the surplus of combustion air, the increase of temperature of combustion air, the supply of inert gas, etc. on the nitrogen oxides emissions of the operating burner under evaluation. Efficient combining of experience and sophisticated approach together with importance of thus access for an improved design are shown.
Current in vitro sonication experiments show immense variability in experimental set-ups and methods used. As a result, there is uncertainty in the ultrasound field parameters experienced by sonicated samples, poor reproducibility of these experiments and thus reduced scientific value of the results obtained. The scope of this narrative review is to briefly describe mechanisms of action of ultrasound, list the most frequently used experimental set-ups and focus on a description of factors influencing the outcomes and reproducibility of these experiments. The factors assessed include: proper reporting of ultrasound exposure parameters, experimental geometry, coupling medium quality, influence of culture vessels, formation of standing waves, motion/rotation of the sonicated sample and the characteristics of the sample itself. In the discussion we describe pros and cons of particular exposure geometries and factors, and make a few recommendations as to how to increase the reproducibility and validity of the experiments performed.
- Klíčová slova
- In vitro experiment, Sonication, Therapeutic ultrasound, Ultrasound exposure,
- MeSH
- analýza selhání vybavení MeSH
- biologické modely MeSH
- buněčné kultury MeSH
- design vybavení MeSH
- počítačová simulace MeSH
- reprodukovatelnost výsledků MeSH
- techniky in vitro MeSH
- ultrazvuková terapie metody MeSH
- vibrace ultrazvukové MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
To study the microvessels in bulbar conjunctiva, we conducted an experiment in goat with a pneumatically driven left heart bypass pump, which was replaced with an undulation pump-left ventricle assist device for 9 days. Three flow patterns were tested: complete pulsatile, continuous, and percentage of pulsatile. We studied the morphology of arterioles and venules of the bulbar conjunctiva using photograph records. The setting up of continuous flow caused global vasoconstriction (significant in venules-P < 0.05). During the pumping in the pulsatile and percentage of pulsatile modes, no significant changes of microvessel morphology were observed. The findings described could point to some disturbances in the microcirculatory bed in conditions of continuous flow.
In this study, we optimized a method for the determination of free amino acids in Nicotiana tabacum leaves. Capillary electrophoresis with contactless conductivity detector was used for the separation of 20 proteinogenic amino acids in acidic background electrolyte. Subsequently, the conditions of extraction with HCl were optimized for the highest extraction yield of the amino acids because sample treatment of plant materials brings some specific challenges. Central composite face-centered design with fractional factorial design was used in order to evaluate the significance of selected factors (HCl volume, HCl concentration, sonication, shaking) on the extraction process. In addition, the composite design helped us to find the optimal values for each factor using the response surface method. The limits of detection and limits of quantification for the 20 proteinogenic amino acids were found to be in the order of 10-5 and 10-4 mol l-1, respectively. Addition of acetonitrile to the sample was tested as a method commonly used to decrease limits of detection. Ambiguous results of this experiment pointed out some features of plant extract samples, which often required specific approaches. Suitability of the method for metabolomic studies was tested by analysis of a real sample, in which all amino acids, except for L-methionine and L-cysteine, were successfully detected. The optimized extraction process together with the capillary electrophoresis method can be used for the determination of proteinogenic amino acids in plant materials. The resulting inexpensive, simple, and robust method is well suited for various metabolomic studies in plants. As such, the method represents a valuable tool for research and practical application in the fields of biology, biochemistry, and agriculture.
- Klíčová slova
- Amino acids, Capillary electrophoresis, Central composite design, Factorial design, Response surface method, Tobacco,
- MeSH
- aminokyseliny izolace a purifikace MeSH
- elektroforéza kapilární metody MeSH
- listy rostlin chemie MeSH
- tabák chemie MeSH
- teoretické modely MeSH
- Publikační typ
- časopisecké články MeSH
- validační studie MeSH
- Názvy látek
- aminokyseliny MeSH
Quantum mechanical (QM) methods have been gaining importance in structure-based drug design where a reliable description of protein-ligand interactions is of utmost significance. However, strategies i. e. QM/MM, fragmentation or semiempirical (SQM) methods had to be pursued to overcome the unfavorable scaling of QM methods. Various SQM-based approaches have significantly contributed to the accuracy of docking and improvement of lead compounds. Parametrizations of SQM and implicit solvent methods in our laboratory have been instrumental to obtain a reliable SQM-based scoring function. The experience gained in its application for activity ranking of ligands binding to tens of protein targets resulted in setting up a faster SQM/COSMO scoring approach, which outperforms standard scoring methods in native pose identification for two dozen protein targets with ten thousand poses. Recently, SQM/COSMO was effectively applied in a proof-of-concept study of enrichment in virtual screening. Due to its superior performance, feasibility and chemical generality, we propose the SQM/COSMO approach as an efficient tool in structure-based drug design.
- Klíčová slova
- in silico drug design, protein-ligand binding, quantum mechanics, semiempirical methods, virtual screening,
- MeSH
- kvantová teorie * MeSH
- ligandy MeSH
- molekulární struktura MeSH
- proteiny chemie MeSH
- racionální návrh léčiv * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- ligandy MeSH
- proteiny MeSH
During the production of mechanically deboned chicken meat (MDCM), a by-product is created that has no adequate use and is mostly disposed of in rendering plants. Due to the high content of collagen, it is a suitable raw material for the production of gelatin and hydrolysates. The purpose of the paper was to process the MDCM by-product into gelatin by 3-step extraction. An innovative method was used to prepare the starting raw material for gelatin extraction, demineralization in HCl, and conditioning with a proteolytic enzyme. A Taguchi design with two process factors (extraction temperature and extraction time) was used at three levels (42, 46, and 50 °C; 20, 40, and 60 min) to optimize the processing of the MDCM by-product into gelatins. The gel-forming and surface properties of the prepared gelatins were analyzed in detail. Depending on the processing conditions, gelatins are prepared with a gel strength of up to 390 Bloom, a viscosity of 0.9-6.8 mPa·s, a melting point of 29.9-38.4 °C, a gelling point of 14.9-17.6 °C, excellent water- and fat-holding capacity, and good foaming and emulsifying capacity and stability. The advantage of MDCM by-product processing technology is a very high degree of conversion (up to 77%) of the starting collagen raw material to gelatins and the preparation of 3 qualitatively different gelatin fractions suitable for a wide range of food, pharmaceutical, and cosmetic applications. Gelatins prepared from MDCM by-product can expand the offer of gelatins from other than beef and pork tissues.
- Klíčová slova
- Taguchi design, biomaterials, by-product, collagen, enzyme conditioning, gelatin, mechanically deboned chicken meat, zero-waste,
- MeSH
- kolagen * chemie MeSH
- kur domácí MeSH
- potraviny MeSH
- skot MeSH
- teplota MeSH
- želatina * chemie MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- kolagen * MeSH
- želatina * MeSH
