Real-time Infection Research Articles

The Role of TIM-1 and CD300a in Zika Virus Infection Investigated with Cell-Based Electrical Impedance.

Orthoflaviviruses cause a major threat to global public health, and no antiviral treatment is available yet. Zika virus (ZIKV) entry, together with many other viruses, is known to be enhanced by phosphatidylserine (PS) receptors such as T-cell immunoglobulin mucin domain protein 1 (TIM-1). In this study, we demonstrate for the first time, using cell-based electrical impedance (CEI) biosensing, that ZIKV entry is also enhanced by expression of CD300a, another PS receptor. Furthermore, inhibiting CD300a in immature monocyte-derived dendritic cells partially but significantly inhibits ZIKV replication. As we have previously demonstrated that CEI is a useful tool to study Orthoflavivirus infection in real time, we now use this technology to determine how these PS receptors influence the kinetics of in vitro ZIKV infection. Results show that ZIKV entry is highly sensitive to minor changes in TIM-1 expression, both after overexpression of TIM-1 in infection-resistant HEK293T cells, as well as after partial knockout of TIM-1 in susceptible A549 cells. These results are confirmed by quantification of viral copy number and viral infectivity, demonstrating that CEI is highly suited to study and compare virus-host interactions. Overall, the results presented here demonstrate the potential of targeting this universal viral entry pathway.

A high-resolution phase-contrast microscopy system for label-free imaging in living cells.

Cell biologists have long sought the ability to observe intracellular structures in living cells without labels. This study presents procedures to adjust a commercially available apodized phase-contrast (APC) microscopy system for better visualizing the dynamic behaviors of various subcellular organelles in living cells. By harnessing the versatility of this technique to capture sequential images, we could observe morphological changes in cellular geometry after virus infection in real time without probes or invasive staining. The tune-up APC microscopy system is a highly efficient platform for simultaneously observing the dynamic behaviors of diverse subcellular structures with exceptional resolution.

Clinical usefulness of linked color imaging in identifying Helicobacter pylori infection: A systematic review and meta-analysis.

Accurate diagnosis of Helicobacter pylori (H. pylori) infection status is a crucial premise for eradication therapy, as well as evaluation of risk for gastric cancer. Recent progress on imaging enhancement endoscopy (IEE) made it possible to not only detect precancerous lesions and early gastrointestinal cancers but also to predict H. pylori infection in real time. As a novel IEE modality, linked color imaging (LCI) has exhibited its value on diagnosis of lesions of gastric mucosa through emphasizing minor differences of color tone. To compare the efficacy of LCI for H. pylori active infection vs conventional white light imaging (WLI). PubMed, Embase, Embase and Cochrane Library were searched up to the end of April 11, 2022. The random-effects model was adopted to calculate the diagnostic efficacy of LCI and WLI. The calculation of sensitivity, specificity, and likelihood ratios were performed; symmetric receiver operator characteristic (SROC) curves and the areas under the SROC curves were computed. Quality of the included studies was chosen to assess using the quality assessment of diagnostic accuracy studies-2 tool. Seven original studies were included in this study. The pooled sensitivity, specificity, positive likelihood rate, and negative likelihood rate of LCI for the diagnosis of H. pylori infection of gastric mucosa were 0.85 [95% confidence interval (CI): 0.76-0.92], 0.82 (95%CI: 0.78-0.85), 4.71 (95%CI: 3.7-5.9), and 0.18 (95%CI: 0.10-0.31) respectively, with diagnostic odds ratio = 26 (95%CI: 13-52), SROC = 0.87 (95%CI: 0.84-0.90), which showed superiority of diagnostic efficacy compared to WLI. Our results showed LCI can improve efficacy of diagnosis on H. pylori infection, which represents a useful endoscopic evaluation modality for clinical practice.

Contact tracing for COVID-19 in a Swiss canton: analysis of key performance indicators.

Contact tracing (CT) has played an important role in strategies to control COVID-19. However, there is limited evidence on the performance of digital tools for CT and no consensus on which indicators to use to monitor their performance. We aimed to describe the system and analyse outcomes of CT with a partially automated workflow in the Swiss canton of Solothurn, using key performance indicators (KPIs). We describe the process of CT used in the canton of Solothurn between November 2020 and February 2022, including forward and backward CT. We developed 16 KPIs representing CT structure (S1-2), process (P1-11) and outcome (O1-3) based on previous literature to analyse the relative performance of CT. We report the changes in the indicators over waves of SARS-CoV-2 infections caused by several viral variants. The CT team in Solothurn processed 57,363 index cases and 71,809 contacts over a 15-month period. The CT team successfully contacted 99% of positive cases within 24 hours (KPI P7) throughout the pandemic and returned almost all test results on the same or next day (KPI P6), before the delta variant emerged. Three-quarters of contacts were notified within 24 hours of the CT interview with the index (KPI P8) before the emergence of the alpha, delta and omicron variants, when the proportions decreased to 64%, 36% and 54%, respectively. The percentage of new symptomatic cases tested and interviewed within 3 days of symptom onset was high at >70% (KPI P10) and contacts started quarantine within a median of 3 days of index case symptom onset (KPI P3). About a fifth of new index cases had already been in quarantine by the time of their positive test (KPI O1), before the delta variant emerged. The percentage of index cases in isolation by day of testing remained at almost 100% throughout the period of analysis (KPI O2). The CT in Solothurn used a partially automated workflow and continued to perform well throughout the pandemic, although the relative performance of the CT system declined at higher caseloads. CT remains an important tool for controlling the spread of infectious diseases, but clearer standards should improve the performance, comparability and monitoring of infection in real time as part of pandemic preparedness efforts.

Animal Models for Toxoplasma gondii Infection.

Toxoplasma gondii is an obligate intracellular protozoan parasite that commonly infects mammals and birds throughout the world. This protocol describes murine models of acute T. gondii infection, toxoplasmic encephalitis and toxoplasma retinochoroiditis. T. gondii infection in severe combined immunodeficient (SCID) mice, deficient in T and B cells, has allowed for the study of T cell-independent mechanisms of defense against intracellular organisms, as described here. The uracil auxotroph strain cps1-1 and temperature-sensitive mutant strains of T. gondii induce protection against challenge with virulent strains of the parasite. They have allowed studies of immunization and adoptive-transfer experiments. A protocol is provided for infection with these mutant strains. The EGS strain of T. gondii has the unique feature of spontaneously forming tissue cysts in cell culture. Dual fluorescent reporter stains of this strain have allowed the study of tachyzoite to bradyzoite transitions in vitro and in vivo. A protocol for in vitro and in vivo growth of this strain and tissue cyst isolation is provided. Genetic manipulation of T. gondii and mice has led to the development of parasites that express fluorescent proteins as well as mice with fluorescently labeled leukocytes. This together with the use of T. gondii that express model antigens and transgenic mice that express the appropriate T cell receptor have facilitated the in vivo study of parasite host-interaction. In addition, parasites that express bioluminescent markers have made it possible to study the dynamics of infection in real time using bioluminescence imaging. Support protocols present methodology for evaluation of progression of infection and immune response to the parasite that includes these newer methodologies. In addition, support protocols address the maintenance of T. gondii tissue cysts and tachyzoites, as well as preparation of T. gondii lysate antigens. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Induction of acute T. gondii infection in mice Basic Protocol 2: Model of toxoplasmic encephalitis and toxoplasma retinochoroiditis in chronically infected mice Basic Protocol 3: Assessment of T. gondii invasion into neural tissue Basic Protocol 4: T. gondii infection in scid/scid (SCID) mice Basic Protocol 5: Infection with the uracil auxotroph strain CPS1-1 or the temperature-sensitive TS-4 strain of T. gondii Basic Protocol 6: In vivo and in vitro maintenance of the EGS strain of T. gondii Support Protocol 1: Assessment of progression of infection and immune response to T. gondii Support Protocol 2: Maintenance of a bank of T. gondii cysts of the ME49 strain Support Protocol 3: Maintenance of T. gondii tachyzoites using human foreskin fibroblasts Support Protocol 4: Maintenance of T. gondii tachyzoites in mice Support Protocol 5: Preparation of T. gondii lysate antigens Support Protocol 6: Isolation of T. gondii tissue cysts from brain.

The Self‐Adaptive Nanosystem for Implant‐Related Infections Theranostics via Phase‐Change Driven Anti‐Biofilm and the Enhancement of Immune Memory

AbstractImplant‐related infections (IRIs), characterized by the formation of bacterial biofilms and immunosuppressive microenvironment, are a common but tricky clinical issue. In this study, a self‐adaptive theranostics nanosystem is designed based on phase change material (PCM) for IRIs therapy, which can monitor the severity of infection in real time and automatically provide the most appropriate treatment on demand. The nanosystem called CaAlg/LOD/POD@MNO/FAs (CLPM) is made of the fatty acids (FAs) core with dissolved minocycline (MNO) and the calcium alginate (CaAlg) shell with an immobilized lactate oxidase/pyruvate oxidase (LOD/POD) enzyme‐linked system. When the level of infection is mild (wound temperature < 39 °C), the CLPMs perform a bacteriostatic function by slowly releasing MNO from the solid‐phase FAs core. When the infection deteriorates (wound temperature > 39 °C), the solid‐to‐liquid phase change of FAs not only enables a rapid release of MNO but also disrupts the mechanical strength of the biofilm skeleton doped with CLPM, performing a bactericidal function. In addition, CLPM can reduce the accumulation of bacteria‐derived lactate to address lactate‐induced immunosuppression and enhance immune memory response, thereby inhibiting infection recurrence.

Monitoring phage-induced lysis of gram-negatives in real time using a fluorescent DNA dye

Bacteriophages (phages) are viruses that specifically attack bacteria. Their use as therapeutics, which constitutes a promising alternative to antibiotics, heavily relies on selecting effective lytic phages against the pathogen of interest. Current selection techniques are laborious and do not allow for direct visualization of phage infection dynamics. Here, we present a method that circumvents these limitations. It can be scaled for high-throughput and permits monitoring of the phage infection in real time via a fluorescence signal readout. This is achieved through the use of a membrane-impermeant nucleic acid dye that stains the DNA of damaged or lysed bacteria and new phage progeny. We have tested the method on Pseudomonas aeruginosa and Klebsiella pneumoniae and show that an increase in fluorescence reflects phage-mediated killing. This is confirmed by other techniques including spot tests, colony plating, flow cytometry and metabolic activity measurements. Furthermore, we illustrate how our method may be used to compare the activity of different phages and to screen the susceptibility of clinical isolates to phage. Altogether, we present a fast, reliable way of selecting phages against Gram-negative bacteria, which may be valuable in optimizing the process of selecting phages for therapeutic use.

Rapid Conductometric Detection of SARS-CoV-2 Proteins and Its Variants Using Molecularly Imprinted Polymer Nanoparticles.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) biosensors have captured more attention than the conventional methodologies for SARS-CoV-2 detection due to having cost-effective platforms and fast detection. However, these reported SARS-CoV-2 biosensors suffer from drawbacks including issues in detection sensitivity, degradation of biomaterials on the sensor's surface, and incapability to reuse the biosensors. To overcome these shortcomings, molecularly imprinted polymer nanoparticles (nanoMIPs) incorporated conductometric biosensor for highly accurate, rapid, and selective detection of two model SARS-CoV-2 proteins: (i) receptor binding domain (RBD) of the spike (S) glycoprotein and (ii) full length trimeric spike protein are introduced. In addition, these biosensors successfully responded to several other SARS-CoV-2 RBD spike protein variants including Alpha, Beta, Gamma, and Delta. Our conductometric biosensor selectively detects the two model proteins and SARS-CoV-2 RBD spike protein variant samples in real-time with sensitivity to a detection limit of 7 pg mL-1 within 10 min of sample incubation. A battery-free, wireless near-field communication (NFC) interface is incorporated with the biosensor for fast and contactless detection of SARS-CoV-2 variants. The smartphone enabled real-time detection and on-screen rapid result for SARS-CoV-2 variants can curve the outbreak due to its ability to alert the user to infection in real time.

A zwitterionic cellulose-based skin sensor for the real-time monitoring and antibacterial sensing wound dressing

Wound infection can induce inflammation and impede wound healing, being a major challenge in wound care management. A household device that can monitor infection in real time, prevent bacterial, and promote wound healing is highly desired but still rarely investigated. In this work, a novel zwitterionic cellulose-based hydrogel that enables continuously real-time monitoring and pro-healing of infected wound is designed. It is based on interpenetrating polymeric networks, including zwitterionic covalent bond network and physical bond network of carboxymethyl cellulose (CMC) with Ag+via metal-coordination interactions. When used as a sensing dressing, it is developed to exhibit pH-responsive and temperature-responsive properties for assessment of wound infection. Moreover, Ag+ can be released from carboxyl groups of CMC in response to the decrease of pH, killing bacteria and promoting wound healing. This zwitterionic carboxymethyl cellulose-based hydrogel sensor opens new avenues for domestic real-time monitoring and pro-healing of infected wound.

Protease inhibitor Camostat Mesyalte blocks wild type SARS-CoV-2 and D614G viral entry in human engineered miniature lungs

The catastrophic global effects of the SARS-CoV-2 pandemic highlight the need to develop novel therapeutics strategies to prevent and treat viral infections of the respiratory tract. To enable this work, we need scalable, affordable, and physiologically relevant models of the human lung, the primary organ involved in the pathogenesis of COVID-19. To date, most COVID-19 in vitro models rely on platforms such as cell lines and organoids. While 2D and 3D models have provided important insights, human distal lung models that can model epithelial viral uptake have yet to be established. We hypothesized that by leveraging techniques of whole organ engineering and directed differentiation of induced pluripotent stem cells (iPSC) we could model human distal lung epithelium, examine viral infection at the tissue level in real time, and establish a platform for COVID-19 related research ex vivo. In the present study, we used type 2 alveolar epithelial cells (AT2) derived from human iPSCs to repopulate whole rat lung acellular scaffolds and maintained them in extended biomimetic organ culture for 30 days to induce the maturation of distal lung epithelium. We observed emergence of a mixed type 1 and type 2 alveolar epithelial phenotype during tissue formation. When exposing our system to a pseudotyped lentivirus containing the spike of wildtype SARS-CoV-2 and the more virulent D614G, we observed progression of the infection in real time. We then found that the protease inhibitor Camostat Mesyalte significantly reduced viral transfection in distal lung epithelium. In summary, our data show that a mature human distal lung epithelium can serve as a novel moderate throughput research platform to examine viral infection and to evaluate novel therapeutics ex vivo.

Remote Health Monitoring in the Workplace for Early Detection of COVID-19 Cases during the COVID-19 Pandemic Using a Mobile Health Application: COVIDApp.

Background: COVIDApp is a platform created for management of COVID-19 in the workplace. Methods: COVIDApp was designed and implemented for the follow-up of 253 workers from seven companies in Catalonia. The assessment was based on two actions: first, the early detection and management of close contacts and potential cases of COVID-19, and second, the rapid remote activation of protocols. The main objectives of this strategy were to minimize the risk of transmission of COVID-19 infection in the work area through a new real-time communication channel and to avoid unnecessary sick leave. The parameters reported daily by workers were close contact with COVID cases and signs and/or symptoms of COVID-19. Results: Data were recorded between 1 May and 30 November 2020. A total of 765 alerts were activated by 76 workers: 127 green alarms (16.6%), 301 orange alarms (39.3%), and 337 red alarms (44.1%). Of all the red alarms activated, 274 (81.3%) were activated for symptoms potentially associated with COVID-19, and 63 (18.7%) for reporting close contact with COVID-19 cases. Only eight workers (3.1%) presented symptoms associated with COVID-19 infection. All of these workers underwent RT-PCR tests, which yielded negative results for SARS-CoV2. Three workers were considered to have had a risk contact with COVID-19 cases; only 1 (0.4%) asymptomatic worker had a positive RT-PCR test result, requiring the activation of protocols, isolation, and contact tracing. Conclusions: COVIDApp contributes to the early detection and rapid activation of protocols in the workplace, thus limiting the risk of spreading the virus and reducing the economic impact caused by COVID-19 in the productive sector. The platform shows the progression of infection in real time and can help design new strategies.

The association between gold mining and malaria in Guyana: a statistical inference and time-series analysis

SummaryBackgroundGuyana reported a significant rise in malaria between 2008 and 2014. As there was no evidence of impairment of national malaria control strategies, public health authorities attributed the surge to a temporal increase in gold mining activity in forested regions. However, systematic analysis of this association is lacking because of the difficulties associated with collecting reliable data for both malaria and mining. We aimed to investigate the association between the international gold price and Plasmodium falciparum malaria transmission in Guyana between 2007 and 2019. We also aimed to evaluate the association between P falciparum cases and the El Niño-Southern Oscillation pattern, which has previously been suggested as a major driver of malaria.MethodsWe used national malaria surveillance data from Guyana to estimate the correlation over time between the international gold price and reported P falciparum infections in individuals who were likely to be involved in mining activities (ie, men and boys aged between 15 and 50 years who were living in mining regions) for each month between 2007 and 2019. We compared the estimates with those obtained from individuals who were unlikely to be directly involved in mining activities (ie, women, children aged 12 years and younger, and adults aged over 70 years) and estimates obtained from individuals living in non-mining regions. We also evaluated the correlation between P falciparum infections and the El Niño-Southern Oscillation pattern in the same subpopulations and time period. Lastly, we evaluated the performance of a statistical model formulated to estimate P falciparum infections in real time using the international gold price as the predictor variable.FindingsThe proportion of P falciparum malaria cases in temporary residents, which was used as a proxy for circulating individuals involved in gold mining, was highest during the years of peak gold price (ie, between 2008 and 2014). Cases of malaria in all demographic groups showed a strong positive correlation with the gold price, but only in regions with mining camps (0·88 [95% CI 0·84–0·89] for boys and men aged between 15 and 50 years and 0·80 [0·73–0·85] for the aggregated population of women, children aged 12 years and younger, and adults older than 70 years). The highest correlation occurred earlier in men and boys aged between 15 and 50 years, the demographic most likely to be miners, suggesting that transmission in mining camps is followed by infections in the community. On the basis of these findings, we were able to reliably forecast P falciparum malaria trends using only the gold price as the predictor variable. A 1% increase in gold price was associated with a 2·13% increase in P falciparum infections after 1 month in the mining populations, and with a 1·63% increase after 2 months in the non-mining populations. Lastly, La Niña climatic events showed an additional, smaller positive correlation with malaria transmission.InterpretationOur analysis provides evidence that the P falciparum malaria surge observed in Guyana between 2008 and 2014 was likely to have been driven mainly by an increase in gold mining, while climate factors might have contributed synergistically. We propose that the international gold price over time is a useful indicator of malaria trends. We conclude that the feasibility of malaria elimination in Guyana, and in other areas in the Amazon where malaria and gold mining overlap, should be evaluated against the challenges posed by rapidly rising gold prices.FundingRamón Areces Foundation, National Institutes of Health, and National Institute of General Medical Sciences.

Current Helicobacter pylori Diagnostics.

The high prevalence of Helicobacter pylori and the variety of gastroduodenal diseases caused by this pathogen necessitate the use of only accurate methods both for the primary diagnosis and for monitoring the eradication effectiveness. There is a broad spectrum of diagnostic methods available for detecting H. pylori. All methods can be classified as invasive or non-invasive. The need for upper endoscopy, different clinical circumstances, sensitivity and specificity, and accessibility defines the method chosen. This article reviews the advantages and disadvantages of the current options and novel developments in diagnostic tests for H. pylori detection. The progress in endoscopic modalities has made it possible not only to diagnose precancerous lesions and early gastric cancer but also to predict H. pylori infection in real time. The contribution of novel endoscopic evaluation technologies in the diagnosis of H. pylori such as visual endoscopy using blue laser imaging (BLI), linked color imaging (LCI), and magnifying endoscopy is discussed. Recent studies have demonstrated the capability of artificial intelligence to predict H. pylori status based on endoscopic images. Non-invasive diagnostic tests such as the urea breathing test and stool antigen test are recommended for primary diagnosis of H. pylori infection. Serology can be used for initial screening and epidemiological studies. The histology showed its value in detecting H. pylori and provided more information about the degree of gastric mucosa inflammation and precancerous lesions. Molecular methods are mainly used in detecting antibiotic resistance of H. pylori. Cultures from gastric biopsies are the gold standard and recommended for antibiotic susceptibility tests.

Active screening of intestinal carbapenem-resistant Enterobacteriaceae in high-risk patients admitted to the hematology wards and its effect evaluation

目的观察血液科肠道碳青霉烯类耐药的肠杆菌科细菌（CRE）主动筛查患者CRE定植、感染的特征，并评价主动筛查结合加强干预在血液科患者CRE感染预防控制中的效果。方法以2017年3月至2019年12月血液内科接受化疗或免疫抑制治疗且预期会出现粒细胞缺乏（粒缺）的患者为研究对象，进行至少3个时间点（治疗前、治疗后粒缺期、粒缺伴发热期）的肠道CRE筛查，以2016年12月至2017年2月血液内科未开展肠道CRE主动筛查的、接受化疗或免疫抑制治疗的115例患者为历史对照组，两组患者均进行CRE感染实时监测，CRE筛查阳性者均采取接触隔离措施，CRE筛查阳性者出现发热或者感染症状时启动针对CRE联合抗生素治疗。结果主动筛查患者CRE定植率为16.46%（66/401）；病种分布上，以急性白血病定植率最高，为23.03%（26/113）。66例筛查阳性患者中，其中第1次筛查阳性患者为27例，占40.9%（27/66），第2次筛查阳性患者为15例，占22.7%（15/66），第3次及以后筛查阳性患者为24例，占36.4%（24/66）。CRE定植病原菌中耐碳青霉烯类肺炎克雷伯菌（CRKP）最多，占54.55%（36/66）。主动筛查患者CRE感染率（2.49%）及死亡率（50.00%）低于历史对照组的11.30%及69.23%；干预期间10例CRE血流感染患者病原菌种类与前期主动筛查病原菌完全相同，符合率为100.0%。结论血液科病房急性白血病患者CRE定植率最高，CRKP是CRE定植、感染的主要病原菌，提高筛查频率可以显著提高筛查阳性率，采取主动筛查并及早干预能有效降低血液科患者CRE发生率及死亡率，CRE筛查阳性病原菌与后续CRE感染病原菌符合率高。恶性血液病患者肠道CRE筛查可以对后期CRE血流感染起到预警以及优化抗菌药物使用的作用。

Microscopic Analysis of Viral Infection.

Viruses engineered to express fluorescent proteins can be used with live-cell imaging techniques to monitor the progression of infection in real time. Here we describe a set of methods to track infection spreading from one cell population to another as well as to visualize transfer of virions between cells. This approach is extended to multiplexing with physiological readouts of cell death, which can be correlated with single-cell resolution to viral infection.

Successfully Sustaining Infection Reductions: A Catheter–Associated Urinary Tract Infection (CAUTI) Prevention Initiative Five Years In

Background: Infection prevention efforts are complex, and sustaining reductions is even more challenging. At the UNC Medical Center, multidisciplinary hospital-wide work groups implement quality improvement initiatives to prevent healthcare-associated infections. The first and most successful initiative has been our catheter-associated urinary tract infection (CAUTI) prevention effort, which started in 2014. The program led to initial dramatic reductions, with continued reductions in CAUTI rates each year since then. Methods: A multidisciplinary workgroup formed in 2014 developed an evidence-based CAUTI prevention bundle and partnered with the nursing staff in 2015–2016 to implement practice changes as part of our hospital’s quality improvement “Spread of Innovations” model. These changes included (1) creation of a 2-person catheter-insertion checklist; (2) insertion skills validation for all nursing staff and nursing assistants; (3) standardization of a maintenance protocol and subsequent education and skills validation with nurses and nurse assistants; and (4) peer audits of urinary catheter maintenance. Additional initiatives implemented over the past 5 years include (1) routine resident education on CAUTI prevention; (2) annual nurse competencies to reinforce skills around CAUTI prevention; (3) introduction of products (eg, PureWick) as alternatives to indwelling catheters; (4) diagnostic stewardship efforts; (5) revisions to the electronic medical record; and (6) efforts to encourage removal of unnecessary catheters such as the “nurse-driven conversation” and Trial of Void. Results: Our CAUTI rates decreased 65% from 2.94 per 1,000 catheter days in the baseline period of 2014 to 1.02 in 2018. In our ICUs (excluding the neonatal ICU), the rate dropped 75% from 4.30 in 2014 to 1.08 per 1,000 catheter days in 2018. Conclusions: We attribute our continued reductions and successful sustainment of low CAUTI rates to several factors. First, the use of a multidisciplinary team was critical to obtaining buy-in from key stakeholders including nursing, nurse assistants, physicians, pharmacists, performance improvement specialists, and administration. Second, continuation of the maintenance peer audits outside the initial project year has provided an important framework for this project, giving regular opportunities for frontline staff to evaluate patients’ catheter condition and to give feedback to colleagues or “just in time education.” These activities potentially prevent infections in real time. Third, with the many competing priorities demanding clinicians’ attention, it has been important for the CAUTI workgroup to continue to evaluate the problem, to determine where opportunities for improvement remain, and to tailor initiatives to meet those needs. In this way, new work can focus on priorities identified by staff, and CAUTI prevention initiatives remain relevant.Funding: NoneDisclosures: None

COVIDApp as an Innovative Strategy for the Management and Follow-Up of COVID-19 Cases in Long-Term Care Facilities in Catalonia: Implementation Study.

BackgroundThe coronavirus disease (COVID-19) pandemic has caused an unprecedented worldwide public health crisis that requires new management approaches. COVIDApp is a mobile app that was adapted for the management of institutionalized individuals in long-term care facilities.ObjectiveThe aim of this paper is to report the implementation of this innovative tool for the management of long-term care facility residents as a high-risk population, specifically for early identification and self-isolation of suspected cases, remote monitoring of mild cases, and real-time monitoring of the progression of the infection.MethodsCOVIDApp was implemented in 196 care centers in collaboration with 64 primary care teams. The following parameters of COVID-19 were reported daily: signs/symptoms; diagnosis by reverse transcriptase–polymerase chain reaction; absence of symptoms for ≥14 days; total deaths; and number of health care workers isolated with suspected COVID-19. The number of at-risk centers was also described.ResultsData were recorded from 10,347 institutionalized individuals and up to 4000 health care workers between April 1 and 30, 2020. A rapid increase in suspected cases was seen until day 6 but decreased during the last two weeks (from 1084 to 282 cases). The number of confirmed cases increased from 419 (day 6) to 1293 (day 22) and remained stable during the last week. Of the 10,347 institutionalized individuals, 5,090 (49,2%) remained asymptomatic for ≥14 days. A total of 854/10,347 deaths (8.3%) were reported; 383 of these deaths (44.8%) were suspected/confirmed cases. The number of isolated health care workers remained high over the 30 days, while the number of suspected cases decreased during the last 2 weeks. The number of high-risk long-term care facilities decreased from 19/196 (9.5%) to 3/196 (1.5%).ConclusionsCOVIDApp can help clinicians rapidly detect and remotely monitor suspected and confirmed cases of COVID-19 among institutionalized individuals, thus limiting the risk of spreading the virus. The platform shows the progression of infection in real time and can aid in designing new monitoring strategies.

COVID-SAFE: An IoT-Based System for Automated Health Monitoring and Surveillance in Post-Pandemic Life.

In the early months of the COVID-19 pandemic with no designated cure or vaccine, the only way to break the infection chain is self-isolation and maintaining the physical distancing. In this article, we present a potential application of the Internet of Things (IoT) in healthcare and physical distance monitoring for pandemic situations. The proposed framework consists of three parts: a lightweight and low-cost IoT node, a smartphone application (app), and fog-based Machine Learning (ML) tools for data analysis and diagnosis. The IoT node tracks health parameters, including body temperature, cough rate, respiratory rate, and blood oxygen saturation, then updates the smartphone app to display the user health conditions. The app notifies the user to maintain a physical distance of 2 m (or 6 ft), which is a key factor in controlling virus spread. In addition, a Fuzzy Mamdani system (running at the fog server) considers the environmental risk and user health conditions to predict the risk of spreading infection in real time. The environmental risk conveys from the virtual zone concept and provides updated information for different places. Two scenarios are considered for the communication between the IoT node and fog server, 4G/5G/WiFi, or LoRa, which can be selected based on environmental constraints. The required energy usage and bandwidth (BW) are compared for various event scenarios. The COVID-SAFE framework can assist in minimizing the coronavirus exposure risk.

From the Cradle to the Grave of an Infection: Host-Pathogen Interaction Visualized by Intravital Microscopy.

During infections, interactions between host immune cells and the pathogen occur in distinct anatomical locations and along defined time scales. This can best be assessed in the physiological context of an infection in the living tissue. Consequently, intravital imaging has enabled us to dissect the critical phases and events throughout an infection in real time in living tissues. Specifically, advances in visualizing specific cell types and individual pathogens permitted tracking the early events of tissue invasion of the pathogen, cellular interactions involved in the induction of the immune response as well the events implicated in clearance of the infection. In this respect, two vantage points have evolved since the initial employment of this technique in the field of infection biology. On the one hand, strategies acquired by the pathogen to establish within the host and circumvent or evade the immune defenses have been elucidated. On the other hand, analyzing infections from the immune system's perspective has led to insights into the dynamic cellular interactions that are involved in the initial recognition of the pathogen, immune induction as well as effector function delivery and immunopathology. Furthermore, an increasing interest in probing functional parameters in vivo has emerged, such as the analysis of pathogen reactivity to stress conditions imposed by the host organism in order to mediate clearance upon pathogen encounter. Here, we give an overview on recent intravital microscopy findings of host-pathogen interactions along the course of an infection, from both the immune system's and pathogen's perspectives. We also discuss recent developments and future perspectives in extracting intravital information beyond the localization of pathogens and their interaction with immune cells. Such reporter systems on the pathogen's physiological state and immune cell functions may prove useful in dissecting the functional dynamics of host-pathogen interactions. © 2019 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

Intravital Imaging of Vaccinia Virus-Infected Mice.

Intravital multiphoton microscopy (MPM) allows the direct visualization of viral infections in real time as they occur in living animals. Here we describe the routes and considerations for murine infection with vaccinia virus (VACV) for imaging, and the preparation of the skin and inner lip (labial mucosa) of infected animals for MPM. Using different recombinant VACVs expressing fluorescent proteins in combination with transgenic fluorescent reporter mice, MPM imaging can be used to examine the movements, interactions, and functions of virus-infected cells or selected immune cell populations after infection.