A handheld sensor assay for the identification of grouper as a safeguard against seafood mislabeling fraud

Increases in international trade and global seafood consumption, along with fluctuations in the supply of different seafood species, have resulted in fraudulent product mislabeling. Grouper species, due to their high demand and varied commercial availability, are common targets for mislabeling by exploiting inefficient inspection practices. Compounding this problem is the fact that there are currently 64 species of fish from eleven different genera allowed to be labeled “grouper” per U.S. Food and Drug Administration guidelines. This wide diversity makes it difficult for regulators to discern legally salable groupers from restricted species. To obviate taxonomic misidentification when relying on external phenotypic characteristics, regulatory agencies are now employing genetic authentication methods which typically offer species-level resolution. However, standard genetic methods such as DNA barcoding require technical expertise and long turnover times, and the required instrumentation is not amenable for on-site analysis of seafood. To obviate some of these limitations, we have developed a handheld genetic sensor that employs a real-time nucleic acid sequence-based amplification assay (RT-NASBA) previously devised in our lab, for the analysis of fish tissue in the field. The base RT-NASBA assay was validated using a lab-based, benchtop RNA purification method as well as non-portable, commercial RT-NASBA analyzer. Described herein, is an uncomplicated method for purifying RNA from fish tissue in the field, which had similar efficiency to the benchtop method demonstrated through direct comparisons. We have also demonstrated that the field sensor is only slightly less sensitive than the benchtop instrument, and could discern 80.3% of groupers (no target sequence available for three species) on the 2014 FDA Seafood List from potential impostors. The complete field assay requires fewer than 80 min for completion and can be performed outside of the lab in its entirety.     

Vital-Fluorochromization of Microorganisms Using 3′,6′-Diacetylfluorescein to Determine Damages of Cell Membranes and Loss of Metabolic Activity by Ozonation

Cell suspensions of Escherichia coli as a model for bacterial populations in wastewaters were treated with ozone as a disinfectant in a continuously operated pilot plant with a plug flow reactor. Suspensions with an initial number of CFU (colony forming units) of 10⁸ mL^?1 were ozonized with ozone concentrations up to 12 mg/L. Metabolic activities and membrane functions break down with increasing ozone concentrations. The fluorochromization using 3′,6′-diacetylfluorescein (FDA) proved to be a suitable method for the detection of an alteration in permeability of the cell membranes and an inactivation of metabolic activity. By fluorescence microscopic and photometric investigations it could be clearly demonstrated that, in the case of disinfection with ozone, inactivation of the cells includes first of all a damage of the cell membranes. In contrast to the determination of the number of CFU, fluorochromization allows the detection of alteration in metabolic activities.     

Virtual Screening of FDA-Approved Drugs against Triose Phosphate Isomerase from Entamoeba histolytica and Giardia lamblia Identifies Inhibitors of Their Trophozoite Growth Phase

Infectious diseases caused by intestinal protozoan, such as Entamoeba histolytica (E. histolytica) and Giardia lamblia (G. lamblia) are a worldwide public health issue. They affect more than 70 million people every year. They colonize intestines causing primarily diarrhea; nevertheless, these infections can lead to more serious complications. The treatment of choice, metronidazole, is in doubt due to adverse effects and resistance. Therefore, there is a need for new compounds against these parasites. In this work, a structure-based virtual screening of FDA-approved drugs was performed to identify compounds with antiprotozoal activity. The glycolytic enzyme triosephosphate isomerase, present in both E. histolytica and G. lamblia, was used as the drug target. The compounds with the best average docking score on both structures were selected for the in vitro evaluation. Three compounds, chlorhexidine, tolcapone, and imatinib, were capable of inhibit growth on G. lamblia trophozoites (0.05–4.935 μg/mL), while folic acid showed activity against E. histolytica (0.186 μg/mL) and G. lamblia (5.342 μg/mL).     

基于FDA的居民用户空调用电行为分类分析方法

针对居民空调用电行为分类中存在事件型数据,导致分类分析耗时长、结果不准确等问题,提出一种基于函数型数据分析（FDA）模型的居民空调用电行为分类分析方法。该方法采用多重分形理论提取居民用电行为特征,使用函数型数据分析算法对居民空调用电行为进行聚类后获取居民空调用电行为类别,采用改进动态时间规整算法对居民空调用电行为实施分类处理,得到居民空调用电行为。根据非介入式设备采集到的实际居民用电行为信息检验该方法的有效性,实验结果表明:该方法可以较好地提取居民用电行为特征,且可有效提高用户空调用电行为分类精度以及缩短分类耗时,可充分描述居民空调开启情况以及消耗电量,具备较好的应用效果。     

基于加权对称图像的二维FDA人脸识别算法

提出一种融合加权对称图像的二维FDA人脸识别算法。将人脸图像分解为奇偶对称脸,并利用加权因子将奇偶对称脸重构新的人脸样本,通过二维FDA算法求解新样本图像的最优特征子空间进行人脸分类。有效融合二维FDA算法的优点,并利用人脸对称性的特征,同时进一步分析加权因子对人脸识别效果的影响,通过选取最优加权因子最大地提高识别率。在人脸图像库ORL中进行的实验结果表明,该算法有效并能获得较高的识别率。     

Genetic algorithms combined with discriminant analysis for key variable identification

Many trouble-shooting problems in process industries are related to key variable identification for classifications. The contribution charts, based on principal component analysis (PCA), can be applied for this purpose. Genetic algorithms (GAs) have been proposed recently for many applications including variable selection for multivariate calibration, molecular modeling, regression analysis, model identification, curve fitting, and classification. In this paper, GAs are incorporated with Fisher discriminant analysis (FDA) for key variable identification. GAs are used as an optimization tool to determine variables that maximize the FDA classification success rate for two given data sets. GA/FDA is a proposed solution for the variable selection problem in discriminant analysis. The Tennessee Eastman process (TEP) simulator was used to generate the data sets to evaluate the correctness of the key variable selection using GA/FDA, and the T² and Q statistic contribution charts. GA/FDA correctly identifies the key variables for the TEP case studies that were tested. For one case study where the correlation changes in two data sets, the contribution charts incorrectly suggest that the operating conditions are similar. On the other hand, GA/FDA not only determines that the operating conditions are different, but also identifies the key variables for the change. For another case study where many key variables are responsible for the changes in the two data sets, the contribution charts only identifies a fraction of the key variables, while GA/FDA correctly identifies all of the key variables. GA/FDA is a promising technique for key variable identification, as is evidenced in successful applications at The Dow Chemical Company.     

The face of the patent is not the “Whole Story”: determining effective life of a pharmaceutical patent in the United States

This article provides an overview of the factors that may contribute to the effective term of protection for a pharmaceutical product in the USA––by patent and by FDA market exclusivities, identifies public and commercial sources for collecting relevant patent term and exclusivity data, and provides a strategy for ensuring that the effective term of protection has been calculated accurately.     

Optimisation of immobilisation conditions for chick pea β-galactosidase (CpGAL) to alkylamine glass using response surface methodology and its applications in lactose hydrolysis

Response surface methodology was advantageously used to optimally immobilise a β-galactosidase from chick pea onto alkylamine glass using Box–Behnken experimental design, resulting in an overall 91% immobilisation efficiency. Analysis of variance was performed to determine the adequacy and significance of the quadratic model. Immobilised enzyme showed a shift in the optimum pH; however, optimum temperature remained unaffected. Thermal denaturation kinetics demonstrated significant improvement in thermal stability of the enzyme after immobilisation. Galactose competitively inhibits the enzyme in both soluble and immobilised conditions. Lactose in milk whey was hydrolysed at comparatively higher rate than that of milk. Immobilised enzyme showed excellent reusability with retention of more than 82% enzymatic activity after 15 uses. The immobilised enzyme was found to be fairly stable in both dry and wet conditions for three months with retention of more than 80% residual activity.     

Butanol isomer separation using polyamide-imide/CD mixed matrix membranes via pervaporation

Mixed matrix membranes (MMMs) comprising polyamide-imide (PAI) and α-, β- or γ-cyclodextrin (CD) have been investigated experimentally and computationally for isomeric n-butanol/tert-butanol (n-BuOH/t-BuOH) separation via pervaporation. Consistent with molecular simulation, experimental results show that the CD inclusion ability and butanol discrimination ability are dependent on both CD cavity size and butanol molecular size. The PAI membrane incorporated with α-CD has the smallest cavity and has the highest discrimination ability for the n-BuOH/t-BuOH pair but with a low butanol flux. The mixed matrix membrane embedded with γ-CD has the lowest selectivity and the highest flux. The PAI/β-CD membrane has a comparable selectivity and flux, and exhibits preferential sorption and diffusion selectivity toward n-BuOH. A maximum separation factor of 1.53 with a corresponding flux of 4.4 g/m² h are obtained at an optimal β-CD loading of 15 wt%. Further increments in the CD content eventually lead to a decrease in separation performance because of CD agglomeration and severe phase separation. To better understand the influence of CD on the separation performance of mixed matrix membranes, SEM, FTIR and XRD have been employed for membrane characterizations. The effect of n-butanol/t-butanol ratio in the feed composition has also been studied. It is found that both flux and separation factor decrease with increasing n-butanol content in the feed. The decline is attributed to the change in total vapor pressure at the upstream and the mutual drag effect of isomeric butanol molecules.     

Determination of pesticide residues in food matrices using the QuEChERS methodology

The determination of pesticide residues in food matrices is a formidable challenge mainly because of the small quantities of analytes and large amounts of interfering substances which can be co-extracted with analytes and, in most cases, adversely affect the results of an analysis. However, safety concerns require that pesticides of the wide range of chemical properties (including acidic, basic and neutral) should be monitored. Because of the wide variety of food matrices, the sample must initially be cleaned up before final analysis. That is why the analytical chemist is faced with the need to devise new methodologies for determining such residues to be determined in a single analytical run. To accomplish the goal, QuEChERS methodology has been developed. It is a streamlined and effective extraction and cleanup approach for the analysis of diverse analyte residues in food matrices. So far, there have been achieved promising results by liquid or gas chromatography analysis, including pesticides, but also acrylamide, pharmaceuticals and veterinary drugs.