Semirational engineering of Cytophaga hutchinsonii polyphosphate kinase for developing a cost-effective, robust, and efficient adenosine 5'-triphosphate regeneration system.

IMPORTANCE: The adenosine 5'-triphosphate (ATP) regeneration system can significantly reduce the cost of many biocatalytic processes. Numerous studies have endeavored to utilize the ATP regeneration system based on Cytophaga hutchinsonii PPK (ChPPK). However, the wild-type ChPPK enzyme possesses limitations such as low enzymatic activity, poor stability, and limited substrate tolerance, impeding its application in catalytic reactions. To enhance the performance of ChPPK, we employed a semi-rational design approach to obtain the variant ChPPK/A79G/S106C/I108F/L285P. The enzymatic kinetic parameters and the catalytic performance in the synthesis of nicotinamide mononucleotide demonstrated that the variant ChPPK/A79G/S106C/I108F/L285P exhibited superior enzymatic properties than the wild-type enzyme. All data indicated that our engineered ATP regeneration system holds inherent potential for implementation in biocatalytic processes.

Validation of an Alzheimer's disease assessment battery in Asian participants with mild to moderate Alzheimer's disease.

There is a lack of validated tools for assessing Alzheimer's disease (AD) across Asia. This study evaluates the psychometric properties of the Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-Cog), Disability Assessment for Dementia (DAD), and Neuropsychological Test Battery (NTB) in Asian participants. Participants with mild to moderate AD (n=251) and healthy controls (n=51) from Mainland China, Taiwan, Singapore, Hong Kong, and South Korea completed selected instruments at several time points. Test-retest reliability was better than 0.70 for all tests. AD participants performed significantly more poorly than controls on every score. Within the AD group, greater disease severity corresponded to significantly poorer performance. The AD group test performance worsened over time and there was a trend for worse performance in AD compared to healthy controls over time. The ADAS-Cog, DAD, and NTB are reliable, valid, and responsive measures in this population and could be used for clinical trials across Asian countries/regions.

Fluorescence excitation-emission spectroscopy with regional integration analysis for assessment of compost maturity.

Composting of animal manures is believed as an alternative way for directly recycling them in farms, and therefore assessment of compost maturity is crucial for achieving high quality compost. Fluorescence excitation-emission matrices (EEMs) combined with regional integration analysis is presented to assess compost maturity. The results showed that the EEM contours of water-extract organic matter (WEOM) from immature composts exhibited four peaks at excitation/emission (Ex/Em) of 220/340nm, 280/340nm, 220/410nm, and 330/410nm, whereas EEM contour of WEOM from mature composts had only two peaks at Ex/Em of 230/420nm and 330/420nm. Pearson correlation demonstrated that peaks intensity rather than their ratios had a significantly correlation with the common indices assessing compost maturity, whereas the normalized excitation-emission area volumes (Φ(i,n)s) from regional integration analysis had a stronger correlation with the common indices assessing compost maturity than peaks intensity. It is concluded that the Φ(i,n)s from regional integration analysis are more suitable to assess the maturity of compost than the intensities of peaks. Therefore, the fluorescence spectroscopy combined with regional integration analysis can be used as a valuable industrial and research tool for assessing compost maturity, given its high sensitivity and selectivity.

Assessment of the profiling microRNA expression of differentiated and dedifferentiated human adult articular chondrocytes.

MicroRNA has an important role in regulating gene expression during cell differentiation. In this study we identified the expression pattern of microRNA in the differentiated and dedifferentiated chondrocytes. Adult human articular chondrocytes were cultured in monolayer. RNA was isolated from the differentiated chondrocytes (collected after isolation) and the fifth-passage (dedifferentiated) chondrocytes, and subjected to gene expression analysis using microRNA and cDNA microarray analysis. Real-time RT-PCR was also performed to confirm the differentially expressed genes. Furthermore, we integrated microRNA and cDNA microarray data together with computational approaches, such as microRNA gene target prediction algorithms, to reveal the role of microRNAs involved in chondrocyte homeostasis. The results showed a dramatic change in expression of microRNA between the two cell types. Thirteen up-regulated and 12 down-regulated microRNAs were detected in differentiated chondroctes. We also revealed microRNA-gene target pairs potentially involved in dedifferentiation process. Our results revealed novel findings of differential expression of microRNA in dedifferentiation, and microRNA could have an important role in the maintenance of chondrocytes homeostasis. MicroRNA may be a target for cartilage tissue engineering and regenerative medicine.

[Mixed logit model and its application in insurance choices of the new rural cooperative medical system].

OBJECTIVE: To apply mixed logit model for analyzing the data of new rural cooperative medical with suitability and identify the factors affecting the residents choices of insurance mode. METHODS: Hypothesis test of IIA was performed using the mogtest module of Stata10.0 to test the eligibility of the condition. The mixed logit model was established to allow the parameters to vary in the population using SAS9.1 MDC module. RESULTS: The data in this study did not satisfy the IIA assumption (P<0.01), so that the multinomial logit model was not applicable. The adjusted Estrella of the mixed logit model was 0.6658. CONCLUSION: The mixed logit approach does not rely on the restrictive IIA assumption and allows for correlation patterns between choices and individual variation. This approach can help in the determination of the choices in new rural cooperative medical system.

PARAFAC modeling of fluorescence excitation-emission spectra for rapid assessment of compost maturity.

Assessment of compost maturity is crucial for achieving high quality compost in order to guarantee its marketability. In this context, a novel technique that combines fluorescence excitation-emission matrices (EEMs) with parallel factor (PARAFAC) analysis to assess compost maturity is presented. A total of 60 fluorescence EEMs of composts were successfully decomposed into a three-factor model using PARAFAC analysis. Components 1 [excitation/emission (Ex/Em) wavelengths=(230, 330)/410] and 2 [Ex/Em wavelengths=(250, 350)/450] were attributable to humic-like and fulvic-like substances, whereas component 3 [Ex/Em wavelengths=(220, 280)/340] belonged to protein-like substances. Pearson correlation analysis between the common maturity indices and log scores of three components demonstrated that components 1 and 3 are more suitable to assess compost maturity than component 2. These results reveal that EEM-PARAFAC could be applied as a valuable tool for assessing compost maturity, given its high sensitivity and selectivity.