Due to their anatomic, physiologic, developmental, and behavioral characteristics, children are particularly vulnerable to bioterrorism agents. Symptoms associated with most bioterrorism agents can be difficult to differentiate from common childhood illnesses. It is extremely important that emergency clinicians are able to recognize unusual illness patterns that could distinguish a natural outbreak from a bioterrorism attack. Resources available through government agencies and leading pediatric organizations can aid in diagnosis and treatment. This issue reviews the highest-risk bioterrorism agents and provides guidance for diagnosing and managing pediatric patients who have been exposed to these agents. [Points & Pearls is a digest of Pediatric Emergency Medicine Practice]

The plant toxin, abrin, a type-II ribosome inactivating protein, is extremely lethal, the human fatal dose being ~1 μg/kg body weight. Abrin has been classified as an agent for bioterrorism, which is of concern. Conversely, the high toxic property of abrin has been employed in generating immunotoxins, whereas its toxin moiety is conjugated to cell surface marker-specific antibodies for cell-targeted killing. Different cell types exhibit variable levels of sensitivity to abrin toxicity; therefore, adequate knowledge of the molecular mechanism that governs the activity of the protein would be a safeguard. To gain insights into this, two cell lines requiring strikingly different concentrations of abrin for inactivating ribosomes were studied. Employing conjugates of the wild-type and active site mutant of abrin A chain with the ricin B chain, it was found that abrin-induced apoptosis was dependent on inhibition of protein synthesis (PSI) leading to ER-stress in Ovcar-3 cells, but not in KB cells. Abrin was also observed to cause direct DNA damage in KB cells, while in Ovcar-3 cells abrin-induced DNA damage was found to be dependent on caspases. Overall, the study demonstrates that the correlation of abrin-mediated PSI and apoptosis is cell-specific and abrin can induce more than one pathway to cause cell death. © 2018 IUBMB Life, 9999(9999):1-7, 2018.

The possibility of exposure to botulinum neurotoxin (BoNT), a powerful and potential bioterrorism agent, is considered to be ever increasing. The current gold-standard assay, live-mouse lethality, exhibits high sensitivity but has limitations including long assay times, whereas other assays evince rapidity but lack factors such as real-time monitoring or portability. In this study, we aimed to devise a novel detection system that could detect BoNT at below-nanomolar concentrations in the form of a stretchable biosensor. We used a field-effect transistor with a p-type channel and electrodes, along with a channel comprising aligned carbon nanotube layers to detect the type E light chain of BoNT (BoNT/E-Lc). The detection of BoNT/E-Lc entailed observing the cleavage of a unique peptide and the specific bonding between BoNT/E-Lc and antibody BoNT/E-Lc (Anti-BoNT/E-Lc). The unique peptide was cleaved by 60 pM BoNT/E-Lc; notably, 52 fM BoNT/E-Lc was detected within 1 min in the device with the antibody in the bent state. These results demonstrated that an all-carbon nanotube-based device (all-CNT-based device) could be produced without a complicated fabrication process and could be used as a biosensor with high sensitivity, suggesting its potential development as a wearable BoNT biosensor.

Bioterrorism content is too often absent from nursing education post-licensure. According to the Centers for Disease Control and Prevention (CDC), bioterrorism is defined as "the deliberate release of viruses, bacteria, or other germs (agents) used to cause illness or death in people, animals, or plants." The purpose of this scholarly work was to develop and pilot a curriculum on introductory bioterrorism concepts for practicing nurses. A literature search and a review of available resources elucidated pertinent content items on bioterrorism and its associated precautions for nurses. Next, two rounds of expert panel consisting of five members rated the identified content and provided comments to validate the curriculum's content. Based on the results, content items were expanded into a curriculum that was piloted as a four hour in person educational session and nurses in attendance were invited to participate in pre-test and post-test questions. They were also asked for their perceptions of the educational session's usefulness and influence on patient care in case of a bioterrorism event. All respondents strongly agreed that the information presented would be useful to them and would influence their patient care in a bioterrorism event. Further refinement, evaluation, and implementation of the developed curriculum are recommended.

Acknowledging the importance of studies toward the development of measures against terrorism and bioterrorism, this study aims to contribute to the design of new prototypes of potential drugs against smallpox. Based on a former study, 9 synthetic feasible prototypes of selective inhibitors for thymidylate kinase from Variola virus (VarTMPK) were designed, and submitted to molecular docking, molecular dynamics simulations and binding energy calculations. The compounds are simplifications of two more complex scaffolds, with a guanine connected to an amide or alcohol through a spacer containing ether and/or amide groups, formerly suggested as promising for the design of selective inhibitors of VarTMPK. Our study showed that, despite the structural simplifications, the compounds presented effective energy values in interactions with VarTMPK and HssTMPK, and that the guanine could be replaced by a simpler imidazole ring linked to a -NH2 group, without compromising the affinity for VarTMPK. It was also observed that a positive charge in the imidazole ring is important for the selectivity towards VarTMPK, and that an amide group in the spacer does not contribute to selectivity. Finally, prototype 3 was pointed as the most promising to be synthesized and experimentally evaluated.

Uganda is prone to epidemics of deadly infectious diseases and other public health emergencies. Though significant progress has been made in response to emergencies during the past 2 decades, system weaknesses still exist, including lack of a robust workforce with competencies to identify, investigate, and control disease outbreaks at the source. These deficiencies hamper global health security broadly. To address need for a highly competent workforce to combat infectious diseases, the Uganda Ministry of Health established the Public Health Fellowship Program (PHFP), the advanced-level Field Epidemiology Training Program (FETP), closely modeled after the CDC's Epidemic Intelligence Service (EIS) program. The 2-year, full-time, non-degree granting program is the first absolute post-master's FETP in Africa for mid-career public health professionals. Fellows gain competencies in 7 main domains, which are demonstrated by deliverables, while learning through service delivery 80% of the time in the ministry of health. During 2015-2017, PHFP enrolled 3 cohorts of 31 fellows. By January 2018, PHFP had graduated 2 cohorts (2015 and 2016) of 19 fellows. Fellows were placed in 17 priority areas of the ministry of health. They completed 153 projects (including 60 outbreak investigations, 12 refugee assessments, 40 surveillance projects, and 31 applied epidemiologic studies), of which 49 involved potential bioterrorism agents or epidemic-prone diseases. They made 132 conference presentations, prepared 40 manuscripts for peer-reviewed publication (17 published as of December 2017), and produced 3 case studies. Many of these projects have resulted in public health interventions that led to improvements in disease control and surveillance systems. The program has produced 19 issues of ministry of health bulletins. One year after graduation, graduates have been placed in key public health decision-making positions. Within 3 years, PHFP has strengthened global health security through improvement in public health emergency response; identification, investigation and control of outbreaks at their sources; and documentation and dissemination of findings to inform decision making by relevant stakeholders.

Due to their anatomic, physiologic, developmental, and behavioral characteristics, children are particularly vulnerable to bioterrorism agents. Symptoms associated with most bioterrorism agents can be difficult to differentiate from common childhood illnesses. It is extremely important that emergency clinicians are able to recognize unusual illness patterns that could distinguish a natural outbreak from a bioterrorism attack. Resources available through government agencies and leading pediatric organizations can aid in diagnosis and treatment. This issue reviews the highest-risk bioterrorism agents and provides guidance for diagnosing and managing pediatric patients who have been exposed to these agents.

Based on prescriptions filled at external pharmacies, prescription surveillance (PS) in Japan has been reporting the estimated numbers of influenza and varicella patients and people prescribed certain drugs since 2009. Every morning, this system estimates the numbers of patients from the numbers of prescriptions filled nationwide for neuraminidase inhibitors, anti-herpes virus drugs, antibiotic drugs, antipyretic analgesics, and multi-ingredient cold medications. Moreover, it can detect "unexplained" infectious diseases that are not explained as infectious diseases monitored by other surveillance systems. Such "unexplained" infectious diseases might be emerging and re-emerging infectious diseases including bioterrorism attacks, which are reportedly difficult to diagnose, at least in early outbreak stages. To ascertain the system's potential benefits, this study examined schemes to detect "unexplained" infectious diseases using PS information. The numbers of patients prescribed the respective drugs are first regressed on the known infectious diseases, time trends, and dummies for day-of-the-week, holidays, and days following a holiday. Known infectious diseases are defined as covered by the National Official Sentinel Surveillance for Infectious Diseases under the Infection Control Law. After the numbers of patients from PS are compared with the predicted numbers of patients, their probabilities of occurrence are calculated. We examined the system's prospective operation from January 2017 through July 2018. The criterion we used to define aberrations varied, from 0.01 to 10-7. For criteria of 0.01 and 10-7 we found 254 and 15 aberrations, respectively. We confirmed its feasibility and effectiveness.

Francisella tularensis is a Gram-negative, facultative, intracellular bacterial pathogen and one of the most virulent organisms known. A hallmark of F. tularensis pathogenesis is the bacterium's ability to replicate to high densities within the cytoplasm of infected cells in over 250 known host species, including humans. This demonstrates that F. tularensis is adept at modulating its metabolism to fluctuating concentrations of host-derived nutrients. The precise metabolic pathways and nutrients utilized by F. tularensis during intracellular growth, however, are poorly understood. Here, we use systematic mutational analysis to identify the carbon catabolic pathways and host-derived nutrients required for F. tularensis intracellular replication. We demonstrate that the glycolytic enzyme phosphofructokinase (PfkA), and thus glycolysis, is dispensable for F. tularensis SchuS4 virulence, and we highlight the importance of the gluconeogenic enzyme fructose 1,6-bisphosphatase (GlpX). We found that the specific gluconeogenic enzymes that function upstream of GlpX varied based on infection model, indicating that F. tularensis alters its metabolic flux according to the nutrients available within its replicative niche. Despite this flexibility, we found that glutamate dehydrogenase (GdhA) and glycerol 3-phosphate (G3P) dehydrogenase (GlpA) are essential for F. tularensis intracellular replication in all infection models tested. Finally, we demonstrate that host cell lipolysis is required for F. tularensis intracellular proliferation, suggesting that host triglyceride stores represent a primary source of glycerol during intracellular replication. Altogether, the data presented here reveal common nutritional requirements for a bacterium that exhibits characteristic metabolic flexibility during infection.IMPORTANCE The widespread onset of antibiotic resistance prioritizes the need for novel antimicrobial strategies to prevent the spread of disease. With its low infectious dose, broad host range, and high rate of mortality, F. tularensis poses a severe risk to public health and is considered a potential agent for bioterrorism. F. tularensis reaches extreme densities within the host cell cytosol, often replicating 1,000-fold in a single cell within 24 hours. This remarkable rate of growth demonstrates that F. tularensis is adept at harvesting and utilizing host cell nutrients. However, like most intracellular pathogens, the types of nutrients utilized by F. tularensis and how they are acquired is not fully understood. Identifying the essential pathways for F. tularensis replication may reveal new therapeutic strategies for targeting this highly infectious pathogen and may provide insight for improved targeting of intracellular pathogens in general.

Environmental sampling is a critical component of the post decontamination verification process following a bioterrorism event. The current work was performed to produce a less labor-intensive method for processing cellulose sponge-wipes used for sampling areas potentially contaminated with low concentrations (i.e., post-decontamination) of Bacillus anthracis spores. An alternative fast-analysis processing method was compared to the processing protocol validated by the Centers for Disease Control and Prevention (CDC) for the Laboratory Response Network (LRN). Glazed tile coupons (1102 cm2) were inoculated with 50, 500, or 5000 spores of Bacillus thuringiensis subsp. kurstaki (Btk), then sampled with cellulose sponges. Sampling was limited to a 25- by 25-cm area and performed in the same manner as the CDC sampling method. Samples were then processed using either the alternative "Fast Analysis" method or the "CDC method". Three different analysts repeated the tests at each concentration utilizing each method. Mean recoveries, labor time, and potentially hazardous waste produced were compared for the two methods. The mean percent recoveries and standard errors for the samples processed using the "CDC method" were 39.9 ± 6.7, 43 ± 7.6, and 36.8 ± 10.1 for the 5000, 500, and 50 spore loading levels, respectively; compared to 54.2 ± 12.9, 64.2 ± 21.7, and 45.2 ± 8.6 for the "Fast Analysis" method. At each titer tested the "Fast Analysis" method resulted in a statistically significant higher percent recovery. Furthermore, analysts processed samples utilizing the "Fast Analysis" method in less than half the time and generated half as much potentially hazardous waste compared to the "CDC method".