Optimization of enzymatic lysis of epithelial cells for application to differential extraction of forensic sexual assault samples

The separation of sperm from female epithelial cells has been a topic of interest in forensic DNA (deoxyribonucleic acid) analysis since the origin of the field. One of the most needed applications of DNA analysis in the identification of the perpetrator of a sexual assault, as often there is little to no other evidence for identification. The largest hurdle to forensic DNA analysis in these cases is that vaginal or oral swabs from sexual assaults will have a mixture of the victim’s epithelial cells and the perpetrator’s sperm cells. It is well known that the analysis of complex mixtures can be difficult to impossible, especially when there is an added concern of low template DNA. Separating these cell types in the mixture evidence is the best way to avoid the need to deduce these difficult mixtures. Sperm and Epithelial Cells are morphologically different both in cell shape and DNA packaging. Nuclear DNA in epithelial cells are more loosely packaged around histones in a structure called a nucleosome. Sperm DNA is tightly packaged around protamines rather than histones. These DNA packaging differences can be utilized to preferentially lyse sperm and epithelial cells in order to separate them. Traditionally this is done by lysing epithelial cells with sodium dodecyl sulfate (SDS) and proteinase K (PK), separating this epithelial DNA from the sperm by centrifugations and finally lysis of the sperm using dithiothreitol (DTT) which reduces the disulfide bonds in the sperm DNA packaging. This method was developed by Peter Gill in 1985 and is still used by forensic laboratories to date. This differential extraction is very labor intensive and time consuming. This dual-enzyme differential extraction can be performed in roughly one hour, which is highly advantageous with the large amount of backlogged sexual assault cases that forensic laboratories have. This work was undertaken to improve the separation of epithelial DNA from sperm cells in the dual-enzyme differential extraction. Here we found that the DNA carryover into the sperm fraction was due to a combination of an inability to completely separate the non-sperm fraction liquid from the sperm pellet and the decreased efficiency of ZyGEM to fully lyse epithelial cells in clumps. The solution to this problem includes the addition of a wash of the sperm pellet after initial separation of the fractions. This wash step decreased the concentration of epithelial DNA to the point that its detection may only occur with very low concentrations of sperm DNA.2017-11-03T00:00:00

Town of Montville Maine Ordinances

Ordinances Cover: Cell Phone Tower; Floodplain Management; Genetically Modified Organisms; Mobile Home Park; Radioactive Waste; Road Entrance Notifications; Shoreland Zoning; site Plan Review; Sludge Disposal; Subdivision; Wind Turbine Generato

Random number generation on handheld devices for cryptographic applications

Random number generation is important in many fields today. It is particularly important in the field of cryptography when generating nonce values, cryptographic keys, and other data required in many cryptographic applications. The proliferation of small, handheld devices that are typically connected to large networks via a wireless connection requires stringent security. Because it may be easier to attack a pseudorandom number generator than to attack a particular cryptosystem, it is important that the generation of random numbers on handheld devices be as secure as possible. In order for the random number generator to provide good, secure "random" data, it must first be seeded by a value that, itself, possesses the qualities of a good random sequence. This paper explores several potential seed sources that are available on many current handheld devices

Bridging the gap: pracademics in foreign policy

In his seminal work Bridging the Gap: Theory and Practice in Foreign Policy, Alexander George (1993) lamented the great divide between academia and the foreign policymaking community, arguing that greater interaction between scholars and policymakers would produce better policy. We share George's belief that scholars and practitioners each have much to offer the other. In fact, a wide spectrum of opportunities exists for scholars in the field of political science to engage in meaningful public service that can enhance every aspect of their careers, as well as contribute to international understanding

Evaluation of strategies for preservation of microalgae Chlorella

[EN] The biomass obtained from microalgae, such as Chlorella, is used to make dietary products, supplements and pharmaceuticals. However, microalgae are produced very far from consumption places. One of the most usual distribution forms is as a dry product, a process that entails high production costs and leads to the loss of certain nutritional properties. Therefore, the aim of this study was to evaluate alternative preservation strategies for microalgae Chlorella other than dehydration and freezing. To that end, sterilization, acidification, and packaging material were analyzed during 2 months of storage under different temperature and light exposure conditions. The results showed that color was modified considerably by sterilization, regardless of light exposure and type of package, whereas citric acid preserved color, especially at low storage temperatures. Furthermore, the study shows that acidification with 3.5% of citric acid and vacuum packaging are the recommended treatment for microalgae, without the need for cold storage. Practical applicationsStabilization of microalgae Chlorella from production to consumption places could increase the possibilities of commercialization of this product, recently labeled superfood by the UN Food and Agriculture Organization. In order to preserve all their nutritional properties for at least 2 months, acidification with 3.5% of citric acid and vacuum packaging are the recommended treatments, without the need for cold storage.The review of this paper was funded by the Universitat Politècnica de València, SpainCastelló Gómez, ML.; Pariente, G.; Andrés Grau, AM.; Ortolá Ortolá, MD. (2017). Evaluation of strategies for preservation of microalgae Chlorella. Journal of Food Processing and Preservation. 42(2):1-8. doi:10.1111/jfpp.13518S1842

Modelling environmental contamination in hospital single and four-bed rooms

Aerial dispersion of pathogens is recognised as a potential transmission route for hospital acquired infections, however little is known about the link between health care worker (HCW) contacts’ with contaminated surfaces, the transmission of infections and hospital room design. We combine computational fluid dynamics (CFD) simulations of bioaerosol deposition with a validated probabilistic HCW-surface contact model to estimate the relative quantity of pathogens accrued on hands during six types of care procedures in two room types. Results demonstrate that care type is most influential (p<0.001), followed by the number of surface contacts (p<0.001) and the distribution of surface pathogens (p=0.05). Highest hand contamination was predicted during Personal care despite the highest levels of hand hygiene. Ventilation rates of 6 ac.h−1 vs 4 ac.h−1 showed only minor reductions in predicted hand colonisation. Pathogens accrued on hands decreased monotonically after patient care in single rooms due to the physical barrier of bioaerosol transmission between rooms and subsequent hand sanitation. Conversely, contamination was predicted to increase during contact with patients in four-bed rooms due to spatial spread of pathogens. Location of the infectious patient with respect to ventilation played a key role in determining pathogen loadings (p=0.05)

Fluctuation between cigarette smoking and use of electronic nicotine delivery systems: Impact on clozapine concentrations and clinical effect

Unlike with smoking cigarettes, electronic nicotine delivery systems do not cause CYP450 1A2 induction as there is a lack of combustion and polycyclic aromatic hydrocarbon production. Changing to the use of an electronic nicotine delivery system from cigarettes can result in the deinduction of CYP450 1A2 and the increase of certain medication serum concentrations, including clozapine. A case is reported in which the switch from smoking to an electronic nicotine delivery system resulted in increased clozapine serum concentration and constipation, necessitating pharmacologic management. The patient ultimately transitioned back to cigarettes, which resulted in the emergence of psychiatric symptoms. An evaluation of longitudinal serum concentrations and clinical correlation is provided. It is important that patients and health care professionals have knowledge not only about the impact of smoking cigarettes on clozapine metabolism, but also the effects of switching to or from an electronic nicotine delivery system

USDA Food and Nutrient Database for Dietary Studies (FNDDS), 5.0

AbstractThe Food and Nutrient Database for Dietary Studies (FNDDS) is used to code and analyze dietary intakes for the What We Eat In America, National Health and Nutrition Examination Survey (WWEIA, NHANES). To create FNDDS 5.0 for WWEIA, NHANES 2009-2010, data for over 7,200 foods were updated to incorporate changes in the marketplace and information reported by survey participants. The updates include nearly 100 new foods and extensive changes to food descriptions, portions, weights, and recipes. The USDA National Nutrient Database for Standard Reference, Release 24 is the basis for the 65 nutrient values for each FNDDS food. FNDDS 5.0 is available at http://www.ars.usda.gov/ba/bhnrc/fsrg