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Wells, George

Publications and source records attributed to Wells, George.

Intensity of sample processing methods impacts wastewater SARS-CoV-2 whole genome amplicon sequencing outcomes

Wastewater SARS-CoV-2 surveillance has been deployed since the beginning of the COVID-19 pandemic to monitor the dynamics in virus burden in local communities. Genomic surveillance of SARS-CoV-2 in wastewater, particularly efforts aimed at whole genome sequencing for variant tracking and identification, are still challenging due to low target concentration, complex microbial and chemical background, and lack of robust nucleic acid recovery experimental procedures. The intrinsic sample limitations are inherent to wastewater and are thus unavoidable. Here, we use a statistical approach that couples correlation analyses to a random forest-based machine learning algorithm to evaluate potentially important factors associated with wastewater SARS-CoV-2 whole genome amplicon sequencing outcomes, with a specific focus on the breadth of genome coverage. We collected 182 composite and grab wastewater samples from the Chicago area between November 2020 to October 2021. Samples were processed using a mixture of processing methods reflecting different homogenization intensities (HA + Zymo beads, HA + glass beads, and Nanotrap), and were sequenced using one of the two library preparation kits (the Illumina COVIDseq kit and the QIAseq DIRECT kit). Technical factors evaluated using statistical and machine learning approaches include sample types, certain sample intrinsic features, and processing and sequencing methods. The results suggested that sample processing methods could be a predominant factor affecting sequencing outcomes, and library preparation kits was considered a minor factor. Finally, a synthetic SARS-CoV-2 RNA spike-in experiment was performed to validate the impact from processing methods and suggested that the intensity of the processing methods could lead to different RNA fragmentation

60 APPLIED LIFE SCIENCES↗

Tailoring polyvinyl alcohol-sodium alginate (PVA-SA) hydrogel beads by controlling crosslinking pH and time

Abstract Hydrogel-encapsulated catalysts are an attractive tool for low-cost intensification of (bio)-processes. Polyvinyl alcohol-sodium alginate hydrogels crosslinked with boric acid and post-cured with sulfate (PVA-SA-BS) have been applied in bioproduction and water treatment processes, but the low pH required for crosslinking may negatively affect biocatalyst functionality. Here, we investigate how crosslinking pH (3, 4, and 5) and time (1, 2, and 8 h) affect the physicochemical, elastic, and process properties of PVA-SA-BS beads. Overall, bead properties were most affected by crosslinking pH. Beads produced at pH 3 and 4 were smaller and contained larger internal cavities, while optical coherence tomography suggested polymer cross-linking density was higher. Optical coherence elastography revealed PVA-SA-BS beads produced at pH 3 and 4 were stiffer than pH 5 beads. Dextran Blue release showed that pH 3-produced beads enabled higher diffusion rates and were more porous. Last, over a 28-day incubation, pH 3 and 4 beads lost more microspheres (as cell proxies) than beads produced at pH 5, while the latter released more polymer material. Overall, this study provides a path forward to tailor PVA-SA-BS hydrogel bead properties towards a broad range of applications, such as chemical, enzymatic, and microbially catalyzed (bio)-processes.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Pattern-Recognition Algorithm for Locking Laser Frequency

A computer program serves as part of a feedback control system that locks the frequency of a laser to one of the spectral peaks of cesium atoms in an optical absorption cell. The system analyzes a saturation absorption spectrum to find a target peak and commands a laser-frequency-control circuit to minimize an error signal representing the difference between the laser frequency and the target peak. The program implements an algorithm consisting of the following steps: Acquire a saturation absorption signal while scanning the laser through the frequency range of interest. Condition the signal by use of convolution filtering. Detect peaks. Match the peaks in the signal to a pattern of known spectral peaks by use of a pattern-recognition algorithm. Add missing peaks. Tune the laser to the desired peak and thereafter lock onto this peak. Finding and locking onto the desired peak is a challenging problem, given that the saturation absorption signal includes noise and other spurious signal components; the problem is further complicated by nonlinearity and shifting of the voltage-to-frequency correspondence. The pattern-recognition algorithm, which is based on Hausdorff distance, is what enables the program to meet these challenges.

Karayan, Vahag↗

Telemetry Monitoring and Display Using LabVIEW

The Measurement Technology Center of the Instrumentation Section configures automated data acquisition systems to meet the diverse needs of JPL's experimental research community. These systems are based on personal computers or workstations (Apple, IBM/Compatible, Hewlett-Packard, and Sun Microsystems) and often include integrated data analysis, visualization and experiment control functions in addition to data acquisition capabilities. These integrated systems may include sensors, signal conditioning, data acquisition interface cards, software, and a user interface. Graphical programming is used to simplify configuration of such systems. Employment of a graphical programming language is the most important factor in enabling the implementation of data acquisition, analysis, display and visualization systems at low cost. Other important factors are the use of commercial software packages and off-the-shelf data acquisition hardware where possible. Understanding the experimenter's needs is also critical. An interactive approach to user interface construction and training of operators is also important. One application was created as a result of a competative effort between a graphical programming language team and a text-based C language programming team to verify the advantages of using a graphical programming language approach. With approximately eight weeks of funding over a period of three months, the text-based programming team accomplished about 10% of the basic requirements, while the Macintosh/LabVIEW team accomplished about 150%, having gone beyond the original requirements to simulate a telemetry stream and provide utility programs. This application verified that using graphical programming can significantly reduce software development time. As a result of this initial effort, additional follow-on work was awarded to the graphical programming team.

graphical user interface LabVIEW graphical program↗

First Lunar Outpost (FLO) mission objectives

This paper discusses four approaches that could be chosen for determining FLO mission objectives. Most important in this discussion is the exploration of the limits set by varying FLO design objectives. The first approach is the FLO design as a 'first and only' lunar visit. Second, the 'independence' approach, is a FLO design as an independent program separate from any Mars program but having long range lunar activities. Third, the 'synergism' approach, is a FLO design with a broad set of future lunar needs and with a primary adjunct of a strong need to help prepare for the Mars missions. Fourth, the 'Mars Prime' approach, is a FLO design drawn from the modification of Mars designs. If cost is paramount but a lunar return is desired, then a 'first and only' is favored. However, most believe that an evolvable lunar program with strong ties to Mars preparation is preferred over the 'first and only'. More recently, the fourth design approach which calls for designs based on Mars needs and lunar elements based on modified Mars elements is drawing more support. This trade is still to be made.

Dupont, Allan L.↗