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Nomenclature, Chemical Abstracts Service Numbers, Isomer Enumeration, Ring Strain and Stereochemistry: What Does Any of This Have to do with an International Chemical Disarmament and Non-Proliferation Treaty?

The Chemical Weapons Convention (CWC) is an international disarmament and non-proliferation treaty that mandates a comprehensive ban on chemical weapons. This treaty entered-into-force in 1997 and has a nearly universal membership of world states. The CWC provides definitions for what constitutes a chemical weapon and describes chemicals for which verification measures and regulatory oversight are required. These chemicals, which must be declared to the Organisation for the Prohibition of Chemical Weapons (OPCW) for verification purposes, appear in three Schedules within the CWC. The schedules list specific chemical substances and families of chemicals defined by molecular structure descriptions that allow a range of specified variable groups. The text of an international treaty provides a basis for laws and regulatory requirements at international and national levels that must be enacted to comply with treaty obligations. These laws and regulations must account for the “chemistry” that is described in the treaty text, which has been negotiated and agreed upon by world governments. In this paper, we look at some of the chemicals described within the CWC and discuss how atoms and molecules, nomenclature and Chemical Abstracts Service (CAS) numbers, isomer enumeration, ring strain and stereochemistry influence the obligations of States to an international treaty. The material draws upon the authors’ experiences from an interactive “Science for Diplomats” initiative that has provided a forum for engaging with chemical disarmament decision makers to aid in their understanding of fundamental concepts in chemistry that have impact on treaty implementation. We hope this article will spark interest from chemistry students and educators in the provision of science advice for policymakers and provide policymakers with an appreciation of the complexity and importance of understanding the language of chemistry in support of a world free of chemical weapons.

Organic Chemistry, Alkanes/Cycloalkanes, Applicati↗

Enumerating soil biodiversity

Soil is an immense habitat for diverse organisms across the tree of life, but just how many organisms live in soil is surprisingly unknown. Previous efforts to enumerate soil biodiversity consider only certain types of organisms (e.g., animals) or report values for diverse groups without partitioning species that live in soil versus other habitats. Here, we reviewed the biodiversity literature to show that soil is likely home to 59 ± 15% of the species on Earth. We therefore estimate an approximately two times greater soil biodiversity than previous estimates, and we include representatives from the simplest (microbial) to most complex (mammals) organisms. Enchytraeidae have the greatest percentage of species in soil (98.6%), followed by fungi (90%), Plantae (85.5%), and Isoptera (84.2%). Our results demonstrate that soil is the most biodiverse singular habitat. By using this estimate of soil biodiversity, we can more accurately and quantitatively advocate for soil organismal conservation and restoration as a central goal of the Anthropocene.

Science & Technology - Other Topics↗

Toward Common Weakness Enumerations in Industrial Control Systems

Here, the storyline of MITRE’s common weakness enumeration framework illustrates how the security and privacy technical community can collaborate/cooperate with policy makers to advance policy, giving it specifics and filling gaps of technical knowledge to improve security and resilience of critical infrastructure.

42 ENGINEERING↗

Listeria monocytogenes in Stone Fruits Linked to a Multistate Outbreak: Enumeration of Cells and Whole-Genome Sequencing

In 2014, the identification of stone fruits contaminated with led to the subsequent identification of a multistate outbreak. Simultaneous detection and enumeration of L. monocytogenes were performed on 105 fruits, each weighing 127 to 145 g, collected from 7 contaminated lots. The results showed that 53.3% of the fruits yielded L. monocytogenes (lower limit of detection, 5 CFU/fruit), and the levels ranged from 5 to 2,850 CFU/fruit, with a geometric mean of 11.3 CFU/fruit (0.1 CFU/g of fruit). Two serotypes, IVb-v1 and 1/2b, were identified by a combination of PCR- and antiserum-based serotyping among isolates from fruits and their packing environment; certain fruits contained a mixture of both serotypes. Single nucleotide polymorphism (SNP)-based whole-genome sequencing (WGS) analysis clustered isolates from two case-patients with the serotype IVb-v1 isolates and distinguished outbreak-associated isolates from pulsed-field gel electrophoresis (PFGE)-matched, but epidemiologically unrelated, clinical isolates. The outbreak-associated isolates differed by up to 42 SNPs. All but one serotype 1/2b isolate formed another WGS cluster and differed by up to 17 SNPs. Fully closed genomes of isolates from the stone fruits were used as references to maximize the resolution and to increase our confidence in prophage analysis. Putative prophages were conserved among isolates of each WGS cluster. All serotype IVb-v1 isolates belonged to singleton sequence type 382 (ST382); all but one serotype 1/2b isolate belonged to clonal complex 5.

59 BASIC BIOLOGICAL SCIENCES↗

Blueprint: Open Worldwide Application Security Project Top 10 Common Weakness Enumerations for Electric Vehicle Supply Equipment

This report categorizes the top 10 security weaknesses in electric vehicle supply equipment (EVSE) using a similar approach by the Open Worldwide Application Security Project (OWASP) Top 10, and it describes example exploitations, methods for prevention, and related common weakness enumerations (CWEs) for each category. The weaknesses listed in this report could lead to security issues or vulnerabilities in EVSE if they are exploited by an attacker to compromise the confidentiality, integrity, or availability of a system. This report also recommends EVSE industry best practices and remediations for the identified weaknesses to use as guidelines for their secure products and systems.

33 ADVANCED PROPULSION SYSTEMS↗

SEAL /Subnetwork Enumeration And Listing/

SEAL /Subnetwork Enumeration And Listing/ computer program uses combinatorial techniques to generate all of the nonredundant subnetwork configurations derivable from an asymmetrical network or device. This is accomplished by a systematic shorting and opening of accessible terminals to obtain the desired allowable configurations.

Happ, W. W.↗

Finding the complete path and weight enumerators of convolutional codes

A method for obtaining the complete path enumerator T(D, L, I) of a convolutional code is described. A system of algebraic equations is solved, using a new algorithm for computing determinants, to obtain T(D, L, I) for the (7,1/2) NASA standard code. Generating functions, derived from T(D, L, I) are used to upper bound Viterbi decoder error rates. This technique is currently feasible for constraint length K less than 10 codes. A practical, fast algorithm is presented for computing the leading nonzero coefficients of the generating functions used to bound the performance of constraint length K less than 20 codes. Code profiles with about 50 nonzero coefficients are obtained with this algorithm for the experimental K = 15, rate 1/4, code in the Galileo mission and for the proposed K = 15, rate 1/6, 2-dB code.

Onyszchuk, I.↗

Rapid enumeration of viable bacteria by image analysis

A direct viable counting method for enumerating viable bacteria was modified and made compatible with image analysis. A comparison was made between viable cell counts determined by the spread plate method and direct viable counts obtained using epifluorescence microscopy either manually or by automatic image analysis. Cultures of Escherichia coli, Salmonella typhimurium, Vibrio cholerae, Yersinia enterocolitica and Pseudomonas aeruginosa were incubated at 35 degrees C in a dilute nutrient medium containing nalidixic acid. Filtered samples were stained for epifluorescence microscopy and analysed manually as well as by image analysis. Cells enlarged after incubation were considered viable. The viable cell counts determined using image analysis were higher than those obtained by either the direct manual count of viable cells or spread plate methods. The volume of sample filtered or the number of cells in the original sample did not influence the efficiency of the method. However, the optimal concentration of nalidixic acid (2.5-20 micrograms ml-1) and length of incubation (4-8 h) varied with the culture tested. The results of this study showed that under optimal conditions, the modification of the direct viable count method in combination with image analysis microscopy provided an efficient and quantitative technique for counting viable bacteria in a short time.

Non-NASA Center↗

Enumerating Spore-Forming Bacteria Airborne with Particles

A laboratory method has been conceived to enable the enumeration of (1) Cultivable bacteria and bacterial spores that are, variously, airborne by themselves or carried by, parts of, or otherwise associated with, other airborne particles; and (2) Spore-forming bacteria among all of the aforementioned cultivable microbes.

Lin, Ying↗

Enumeration and Fluorescence In Situ Hybridization of Microbial Bioburden on Cleanroom Surfaces

Introduction: Microorganisms are everywhere on Earth, even in the cleanest of places. Spacecraft assembly cleanrooms can harbor low levels of living and dead microbial cells (e.g., [1,2]), and cleanroom bioburden can also include organic molecules from industrial sources and in situ biomass. Life detection missions require careful attention to avoid contaminants that can be easily convoluted with analytical targets. We are evaluating epifluorescent microscopy and fluorescence in situ hybridization (FISH) as methods to complement organic contamination detection techniques. Epifluorescent cell counting offers an accurate and cost-effective way to quantify low levels of surface biomass. FISH could allow for the identification of residual organisms, and can be targeted to detect active populations of specific organisms such as bacteria known to resist cleaning procedures. This effort is part of a larger study that is concentrated on characterizing the surface and airborne molecular organic contamination background in Johnson Space Center (JSC) Astromaterials curation laboratories and Goddard Space Flight Center (GSFC) spacecraft assembly rooms, and understanding contaminants in the context of cleaning procedures and residual bioburden. Methods: Samples were collected by swabbing surfaces in ISO 5 and ISO 7 equivalent cleanrooms at JSC. Swabs for FISH were fixed in 4% paraformaldehyde (PFA) for 3 hours and then stored in 1:1 ethanol:PBS, while swabs for cell counting were stored in 4% PFA until analysis to avoid any cell loss during centrifugation that could impact quantification of very low biomass samples. Cell counting was performed with SYBR Gold as in [3], but adapted for very low biomass. FISH was performed as in [4], using DAPI as a counterstain for all DNA-containing cells. Negative controls included wells with no probe applied, to test for natural fluorescence, as well as the nonsense probe NONEUB (reverse complement of EUB338) to evaluate non-specific probe binding. Results and Discussion: Cleanroom surfaces had 102-103 cells cm-2. The extremely low biomass of these samples was challenging for enumeration, and required careful and routine use of “field” and laboratory blanks. FISH was performed with the general archaeal and bacterial probes ARCH915 and EUB338 (EUBMIX, [4]), probe GAMBET ([4]), and PSE227, which targets the genus Pseudomonas [5]). The latter two probes were selected because Pseudomonas spp. and other Gammaproteobacteria have not been isolated from cleanroom surfaces but do appear frequently in rRNA gene libraries from these surfaces. While some active bacteria were identified (Fig. 1c), most cells detectable by DAPI did not have a strong or any fluorescent signal (e.g., Fig. 1d), indicating that the vast majority of cells are dead or inactive. This suggests that cleaning protocols are effective at inactivating microbial contaminants, but that dead or inactive cells can remain on surfaces. Cells were often clumped in a weakly autofluorescent matrix, possibly biofilm material (Fig. 1c,d). We also observed other particulate material that was collected by the swabs, including apparent textile fibers (Fig. 1b). Our results are consistent with other studies that show that the bioburden present in clean rooms includes active, dormant, and dead cells. We will discuss how FISH and epifluorescent cell counting could be applied in planetary protection protocols, including the advantages and disadvantages of FISH and cell counting for routine use, as well as different possible applications for more specialized FISH procedures. References: [1] Moissl-Eichinger et al. (2015) Sci Rep, 5, 9156 [2] Hendrickson et al. (2021) Microbiome, 9, 238 [3] Jones et al. (2017) Appl Environ Microbiol, 83, e00909-17 [4] Jones et al. (2015) Appl Environ Microbiol, 81, 1242-1250. [5] Watt et al. (2006) Environ Microbiol, 8, 871-884

C J Huff↗