Evaluation of Saccharomyces cerevisiae modified via CRISPR/Cas9 as a cellulosic platform microorganism in simultaneously saccharification and fermentation processes.

The nonrenewable character and deleterious effects of fossil fuels foster the need for cleaner and more inexhaustible energy sources, such as bioethanol. Especially from lignocellulosic biomasses. However, the economic viability of this product in the market depends on process optimization and cost reduction. This research applied a sequential experimental project to investigate the process of enzymatic saccharification and simultaneous fermentation to produce ethanol with sugarcane bagasse. The differential of the work was the application of the strain of Saccharomyces cerevisiae AGY001 which was improved by evolutionary engineering to become thermotolerant and by a heterologous expression based on genomic integration by CRISPR/Cas9 to produce endoglucanase and ß-glucosidase (AsENDO-AsBGL). The maximum ethanol yield found was 89% of the maximum theoretical yield (released sugars), obtained at temperature concentrations, sugarcane bagasse and inoculum at 40 °C, 16.5%, and 4.0 g/L, respectively (12.5 FPU/g bagasse). The mathematical model obtained can predict approximately 83% of the data set with 95% confidence. Therefore, these findings demonstrated the potential of sugarcane bagasse and S. cerevisiae AGY001 strain (CRISPR/Cas9 modified) in bioethanol production without the need for impractical selection media on an industrial scale, in addition to providing useful insights for the development of SSF processes.

Multivariate Analysis Identifying [Cu(N

White light-emitting electrochemical cells (LECs) comprising only [Cu(N^N)(P^P)]+ have not been reported yet, as all the attempts toward blue-emitting complexes failed. Multivariate analysis, based on prior-art [Cu(N^N)(P^P)]+ -based thin-film lighting (>90 papers) and refined with computational calculations, identifies the best blue-emitting [Cu(N^N)(P^P)]+ design for LECs, that is, N^N: 2-(4-(tert-butyl)phenyl)-6-(3,5-dimethyl-1H-pyrazol-1-yl)pyridine and P^P: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, to achieve predicted thin-film emission at 490 nm and device performance of 3.8 cd A-1 @170 cd m-2 . Validation comes from synthesis, X-ray structure, thin-film spectroscopic/microscopy/electrochemical characterization, and device optimization, realizing the first [Cu(N^N)(P^P)]+ -based blue-LEC with 3.6 cd A-1 @180 cd m-2 . This represents a record performance compared to the state-of-the-art tricoordinate Cu(I)-complexes blue-LECs (0.17 cd A-1 @20 cd m-2 ). Versatility is confirmed with the synthesis of the analogous complex with 2-(4-(tert-butyl)phenyl)-6-(3,5-dimethyl-1H-pyrazol-1-yl)pyrazine (N^N), showing a close prediction/experiment match: λ = 590/580 nm; efficiency = 0.55/0.60 cd A-1 @30 cd m-2 . Finally, experimental design is applied to fabricate the best white multicomponent host:guest LEC, reducing the number of trial-error attempts toward the first white all-[Cu(N^N)(P^P)]+ -LECs with 0.6 cd A-1 @30 cd m-2 . This corresponds to approximately ten-fold enhancement compared to previous LECs (<0.05 cd A-1 @<12 cd m-2 ). Hence, this work sets in the first multivariate approach to design emitters/active layers, accomplishing first-class [Cu(N^N)(P^P)]+ -based blue/white LECs that were previously elusive.

A Web-Based Intervention to Increase Smokers' Intentions to Participate in a Cessation Study Offered at the Point of Lung Screening: Factorial Randomized Trial.

BACKGROUND: Screen ASSIST is a cessation trial offered to current smokers at the point of lung cancer screening. Because of the unique position of promoting a prevention behavior (smoking cessation) within the context of a detection behavior (lung cancer screening), this study employed prospect theory to design and formatively evaluate a targeted recruitment video prior to trial launch. OBJECTIVE: The aim of this study was to identify which message frames were most effective at promoting intent to participate in a smoking cessation study. METHODS: Participants were recruited from a proprietary opt-in online panel company and randomized to a 2 (benefits of quitting vs risks of continuing to smoke at the time of lung screening; BvR) × 2 (gains of participating vs losses of not participating in a cessation study; GvL) message design experiment (N=314). The primary outcome was self-assessed intent to participate in a smoking cessation study. Message effectiveness and lung cancer risk perception measures were also collected. Analysis of variance examined the main effect of the 2 message factors and a least absolute shrinkage and selection operator (LASSO) approach identified predictors of intent to participate in a multivariable model. A mediation analysis was conducted to determine the direct and indirect effects of message factors on intent to participate in a cessation study. RESULTS: A total of 296 participants completed the intervention. There were no significant differences in intent to participate in a smoking cessation study between message frames (P=.12 and P=.61). In the multivariable model, quit importance (P<.001), perceived message relevance (P<.001), and affective risk response (ie, worry about developing lung cancer; P<.001) were significant predictors of intent to participate. The benefits of quitting frame significantly increased affective risk response (Meanbenefits 2.60 vs Meanrisk 2.40; P=.03), which mediated the relationship between message frame and intent to participate (b=0.24; 95% CI 0.01-0.47; P=.03). CONCLUSIONS: This study provides theoretical and practical guidance on how to design and evaluate proactive recruitment messages for a cessation trial. Based on our findings, we conclude that heavy smokers are more responsive to recruitment messages that frame the benefits of quitting as it increased affective risk response, which predicted greater intention to participate in a smoking cessation study.

Combination of Catechin, Epicatechin, and Rutin: Optimization of a novel complete antidiabetic formulation using a mixture design approach.

Nowadays, synthetic chemical antidiabetic drugs, besides their therapeutic effects, present adverse effects that could be hard to handle over time. In the last decade, studies reported new alternative molecules with more health benefits and less adverse effects. The goal of this study is to optimize a new antidiabetic formulation using plant flavonoids: Catechin, Epicatechin, and Rutin. They are also a powerful antioxidant and anti-inflammatory molecules. A mixture design experiment will optimize their combination to obtain a new, safe multi-targets antidiabetic formulation making it a powerful combination for the management of diabetes and its complications. To study the variation of blood glucose level in response to the treatment over the time we performed an Oral Glucose Tolerance Test. The blood glucose level variations recorded as responses for the mixture design experiment. We used the molecules at a dose of 10 mg/kg. According to the software analysis, the prediction profiler showed us the optimum combination, and the result was a binary combination between Rutin and Epicatechin (25% and 75%, respectively). This combination prevented hyperglycemia and hypoglycemia, along with the best area under the curve, and after that, we validated it through a repeated oral administration on alloxan-induced diabetic mice for 28 d. Rutin, Catechin, and Epicatechin exhibit a potent antihyperglycemic activity, their synergistic combination validates a new formulation that could be a real candidate to conventional drugs.

[Spray drying process of extract of Wenjing Zhitong Prescription based on mixture design experiment].

To improve the spray drying effect of extract of Wenjing Zhitong Prescription, this study takes the yield, hygroscopic property and the fluidity of dry powder as indexes to screen out auxiliary materials, and the proportion of the auxiliary materials was optimized based on the mixing design experiment; based on that, HPLC method was established for the determination of glycyrrhizin and 6-gingerol in spray powder, the yield of spray powder and the retention rate of the two index components were taken as indexes to further optimize the spray drying parameters. The finally selected auxiliary materials were light magnesium oxide, maltodextrin and silica, and regression equations of dry powder yield, moisture absorption rate, angle of rest with proportion of auxiliary materials were established, and the optimized proportion of auxiliary materials was dry paste-light magnesium oxide-maltodextrin-silica=0.5∶0.305∶0.145∶0.05; according to the optimized drying process parameters of Wenjing Zhitong Prescription, initial temperature was 60 ℃, air inlet temperature was 130 ℃, air flow rate was 35 m~3·h~(-1), atomizing pressure was 40 mm, and liquid inlet speed was 4.5 mL·min~(-1). Under these conditions, the dry powder yield was 90.28%, the retention rate of glycyrrhizin was 74.51%, and the retention rate of 6-gingerol was 72.10%. In this study, optimized auxiliary materials can improve the yield of spray drying and the property of spray powder, and the optimized processing conditions were good for retaining the unstable gingerol components, which can lay a foundation for the further preparation research of meridian warming and pain relieving prescriptions, and provide reference for extract of other traditional Chinese medicine extracts that are difficult to spray drying.

Spray drying process of extract of Wenjing Zhitong Prescription based on mixture design experiment / 中国中药杂志

To improve the spray drying effect of extract of Wenjing Zhitong Prescription, this study takes the yield, hygroscopic property and the fluidity of dry powder as indexes to screen out auxiliary materials, and the proportion of the auxiliary materials was optimized based on the mixing design experiment; based on that, HPLC method was established for the determination of glycyrrhizin and 6-gingerol in spray powder, the yield of spray powder and the retention rate of the two index components were taken as indexes to further optimize the spray drying parameters. The finally selected auxiliary materials were light magnesium oxide, maltodextrin and silica, and regression equations of dry powder yield, moisture absorption rate, angle of rest with proportion of auxiliary materials were established, and the optimized proportion of auxiliary materials was dry paste-light magnesium oxide-maltodextrin-silica=0.5∶0.305∶0.145∶0.05; according to the optimized drying process parameters of Wenjing Zhitong Prescription, initial temperature was 60 ℃, air inlet temperature was 130 ℃, air flow rate was 35 m~3·h~(-1), atomizing pressure was 40 mm, and liquid inlet speed was 4.5 mL·min~(-1). Under these conditions, the dry powder yield was 90.28%, the retention rate of glycyrrhizin was 74.51%, and the retention rate of 6-gingerol was 72.10%. In this study, optimized auxiliary materials can improve the yield of spray drying and the property of spray powder, and the optimized processing conditions were good for retaining the unstable gingerol components, which can lay a foundation for the further preparation research of meridian warming and pain relieving prescriptions, and provide reference for extract of other traditional Chinese medicine extracts that are difficult to spray drying.

Optimization of nimesulide-loaded solid lipid nanoparticles (SLN) by factorial design, release profile and cytotoxicity in human Colon adenocarcinoma cell line.

The aim of this work is development of a nontoxic, long-term stable solid lipid nanoparticles (SLN) formulation for the loading of Nimesulide (NiM) by a 22 factorial design. The optimized formulation was composed of 10 wt% of glyceryl behenate and 2.5 wt% of poloxamer 188. Immediately after production, Z-Ave of NiM-SLN was 166.1 ± 0.114 nm, with a polydispersity index (PI) of 0.171 ± 0051 and zeta potential nearly neutral (-3.10 ± 0.166 mV). A slight increase of Z-Ave was recorded for NiM-SLN stored at 25 °C for a period of 15 days, whereas at 4 °C particles kept size within similar range. Long-term stability was monitored using TurbiscanLab®, showing a high stability of the nanoparticles with variations in the backscattering profiles below 10%. The release profile of NiM-SLN followed a sustained pattern with ca. 30% of drug released up to 24 h. Empty-SLN and NiM-SLN were nontoxic after exposing Caco-2 cells to the highest concentration (100 µg/mL) up to 48 hours (cell viability higher than 80%). NiM-SLN were lyophilized using different cryoprotectants, producing particles of 463.1 ± 36.63 nm (PI 0.491 ± 0.027) with 5% trehalose. Solid character of NiM-SLN was confirmed by DSC, recording a recrystallization index of 83% for NiM-SLN and of 74% for lyophilized SLN.

"The application effect of""comprehensive design experiment evaluation""in the evaluation of nursing experimental teaching" / 中华医学教育探索杂志

Objective To explore the value of applyingcomprehensive design experiment evalua-tionin the evaluation of nursing experimental teaching. Methods In the nursing profession, and after the acute and severe cases were collected, comprehensive design experiment evaluation was carried out by using the first-aid and severe training programs, according to the patients needs. Then the simulation effect of students was evaluated by multi-subjects Afterwards, a questionnaire survey was conducted among 143 participants on clinical competency and experimental assessment. The data were analyzed through ratio. Result 121 nursing students' (84.6%) clinical competence was improved. 143 nursing students (100%) thought that the test was of great significance to achieve education target. 138 nursing students (96.6%) thought that general design and arrangement of satisfaction were high. 141 nursing students (98.7%) were satisfied with the multi-subject evaluation. 106 nursing students (74.1%) were willing to conduct self evaluation and give evaluation of others. Conclusion Comprehensive designing experiments evaluationcan significantly improve the critical clinical competence of nursing students, and can help the professional education target implementation. The operation mode is recognized by the students. The evaluation of the multi-subject evaluation is highly respected, but it needs to develop the students' ability of objective evalua-tion.

Innovating Experimental Teaching System and Improving Students’ Practicing Ability——Experimental Teaching Reformation of Environmental Microbiology / 微生物学通报

A new experiment curriculum system of environmental microbiology was established centering on applied microbiology in environmental protection field and emphasizing on design and research experi- ments to motivate the students’ interests for the course, which helped them to improve their ability of think- ing independently and creatively as well as their practicing ability.

The exploration of the student-centered comprehensive and designing experiment / 中华医学教育探索杂志

As an important method,the experimental teaching can enhance the compe-tence of students such as operation ability,and make students learn how to solve the problem scientifically and innovatively.The aim of comprehensive experiment is to make studetns develop fully and understand the relationship between theory and practice.The designing experiment is to training students to complete the whole experiment process by themselves from reading reference to write the paper.All these trainings intend to improve the scientific research ability of under-graduate students.

The practice and discussion of"six processes"teaching methods in functional design experiments / 中华医学教育探索杂志

The functional experiment teaching is an inspection to students of their thinking methods and the operation abilities,and is also the summary and sublimation of this course theories teaching.To adapt to the demand for cultivation of the talented person by innovation education,we probe into the improvement of teaching methods of the functional design experiment,and practiced the new teaching system of "six processes"teaching methods such as speaking briefly,giving a demonstration,questioning,giving clue,study and discussion,evaluation,and push forward the biomedical science modes toward the development of "living creature-mental state-social medical science mode"direction.

The teaching method of medical function design experiment / 中华医学教育探索杂志

The course of medical function design experiment can improve students’inter-ests,strengthen their practice ability and the ability to apply the knowledge synthetically,and de-velop their creativity consciousness and research ability. This article explores the teaching method of design experiment and its function in teaching.

Learning Theory as Teaching Resource: Enhancing Students' Understanding of Economic Concepts.

A group of experienced secondary school teachers used a novel learning theory as a resource for planning and carrying out their teaching of a difficult economic concept. Their students' mastery of this concept after a series of three lessons was compared with the mastery of the same concept by students who were taught by another group of teachers under the same conditions except for the use of the theory. The difference in learning outcomes was extreme. Observations of what was happening in the classrooms showed subtle but decisive differences correlated with the differences in outcome. These differences were interpreted in terms of the theory used by the first group, and the results seem to give support to the theoretical claim that for any specific object of learning there is a necessary pattern of variation and invariance that the learners must experience in order to appropriate the object of learning in question and thus by bringing out that pattern in the learning situation, the likelihood of that object of learning being appropriated is enhanced. Furthermore, this study shows how the understanding of the simultaneous change in the supply of and the demand for a certain good affects its market price can be brought about in a powerful way.

Technical support function in the independent proposition design of the experimental teaching implementation / 中华医学教育探索杂志

The design experiment carrying out requests higher demands for the technical preparations work.How to do the technical support work well in large-scale design experiments is a worth exploring question.The author accumulates some experiences and appreciates from experience according to design experiment practice carried out in several underrates,hoping to exchange ideas with the peers.