Fatal Infection Research Articles

Outcomes of Adjunctive Therapy with Open Arthrotomy for Patients with Septic Arthritis Due to Melioidosis: A Retrospective Study.

Melioidosis is a potentially fatal infection caused by the bacterium Burkholderia pseudomallei. Septic arthritis caused by this infection is uncommon and difficult to treat. The role of adjunctive open arthrotomy in this type of infection has not yet been elucidated. We conducted a retrospective study of patients with microbiologically confirmed melioidosis between January 2002 and December 2022. Patients with a clinical condition of septic arthritis and positive cultures for B. pseudomallei were included. Comparisons were made between patients who received adjunctive therapy with open arthrotomy with conventional standard treatment and those who did not in terms of clinical outcomes and hospital expenditures. Of the 478 patients diagnosed with melioidosis microbiological confirmation, 81 patients had septic arthritis, accounting for 17% of cases. Among these patients, only 36 (44%) underwent adjunctive therapy with open arthrotomy. The 14-day and 30-day in-hospital mortality and length of hospital stays of patients who underwent adjunctive therapy with open arthrotomy were more favorable than those of patients who did not receive adjunctive therapy with open arthrotomy; however, the difference was not statistically significant. Patients who underwent adjunctive therapy with open arthrotomy had lower hospital expenditures (antimicrobial and non-antimicrobial costs) than those who did not undergo open arthrotomy. Adjunctive therapy with open arthrotomy for patients with septic arthritis due to melioidosis was associated with favorable clinical outcomes and significantly lower hospital expenditures.

Production and characterization of biologicals for disease diagnosis and pathological evaluation of elephant endotheliotropic herpesvirus (EEHV)

Elephant endotheliotropic herpesviruses (EEHV) belong to the family Herpesviridae and cause a highly fatal hemorrhagic infection in elephants. EEHV poses a global threat to the already endangered elephant population. Since EEHV is a non-cultivable virus, there is a scarcity of specific diagnostics, therapeutics, and vaccines. In this study, our objective was to develop biologicals for diagnosis and pathological studies against the most prevalent EEHV1A/1B. We expressed two truncated fragments of the DNA polymerase, glycoprotein B (gB), and glycoprotein (gL) of EEHV in the prokaryotic system. Hyperimmune serum against the purified antigens was raised in rabbits and guinea pigs. We validated the reactivity of this hyperimmune serum using western blotting, ELISA, and immune-histochemistry on known positive infected tissues. Samples collected from 270 animals across various states in India were evaluated with these biologicals. The raised antibodies successfully demonstrated virus in immune-cytochemistry. Additionally, all known positive samples consistently exhibited significant inhibition in the OD values when used in the competitive format of ELISA across all four antigens when compared to the serum collected from known negative animals. An apparent sero-prevalence of 10 % was observed in the randomly collected samples. In summary, our study successfully developed and validated biologicals that will be invaluable for EEHV diagnosis and control.

Incidence of infections in clinical trials of treatments for relapsed/refractory multiple myeloma: Systematic review and meta-analysis.

e19517 Background: Although immunotherapy shows favorable survival outcomes among heavily pretreated relapsed/refractory multiple myeloma (RRMM) patients, infections are a common cause of morbidity and mortality that are not well characterized in this patient population. The goal of this study was to estimate the incidence of infections from clinical trials of treatments for RRMM by grade, infection type, and treatment received. Methods: We conducted a literature search in EMBASE, PubMed, Cochrane, approval packages and labeling, and conference proceedings to identify phase 2-3 clinical trials of single drug interventions published prior to 18 June 2023 with data on infections. We included studies of patients aged ≥18 years who received prior MM treatment with an immunomodulatory agent, protease inhibitor, and anti-CD38 antibody. A Bayesian meta-analysis accounting for left-censored data was used to estimate the pooled incidence of infection from the studies by infection type (upper respiratory tract infection [uRTI], sepsis, bacteremia, pneumonia, SARS-COV-2, and bacterial and viral infections), grade (all, grades ≥3, and 5/fatal) and type of treatment received (antibody and CAR-T). Results: Nine studies (7 phase 2, 1 phase 3 and 1 product label) of seven compounds (1 selective inhibitor of nuclear export, 2 CAR-T, and 4 monoclonal and bispecific antibodies) were identified and are/were approved in the US for RRMM treatment. All included articles were published after July 2019 on treatments approved since 2020 for patients who had received ≥4 prior lines of therapy. They included a total of 1,141 patients (median follow-up 10.1 - 23 months). The pooled incidence of all infections was 61.6% (95% credible interval, 58.6%-64.6%), 28.7% (24.7%-32.2%) and 4.1% (2.9%-5.6%) for all-grades, grades ≥3, and 5 respectively. uRTI was the most common type of all-grade infection (22.8% [20.0%-25.6%]) while SARS-COV-2 (6.8% [4.9%-9.4%]) and pneumonia (6.8% [5.0%-9.1%]) were the most common grade ≥3 infections. Sepsis was the most common fatal infection (1.0% [0.4%-1.8%]). Patients treated with monoclonal/bispecific antibodies had a slightly higher incidence of all-grade infections (64.2% [60.4%-67.9%]) compared to CAR-T (58.6% [52.8%-64.2%]). The incidences for these treatments were similar for grades ≥3 (28.9% vs 28.2%) and 5 (3.5% vs 4.5%) infections. Conclusions: In clinical trials of treatment for triple class exposed RRMM, the incidence of infection was high, including grade 3 or higher infections. A large proportion of grade ≥3 infections were attributable to SARS-COV-2 and pneumonia, with sepsis as the most common fatal infection. The incidence of grade ≥3 infection was generally consistent for CAR-T and monoclonal/bispecific antibody therapy.

Mutation of hmgA, encoding homogentisate 1,2-dioxygenase, is responsible for pyomelanin production but does not impact the virulence of Burkholderia cenocepacia in a chronic granulomatous disease mouse lung infection.

The Burkholderia cepacia complex (Bcc) is a group of Gram-negative opportunistic bacteria often associated with fatal pulmonary infections in patients with impaired immunity, particularly those with cystic fibrosis (CF) and chronic granulomatous disease (CGD). Some Bcc strains are known to naturally produce pyomelanin, a brown melanin-like pigment known for scavenging free radicals; pigment production has been reported to enable Bcc strains to overcome the host cell oxidative burst. In this work, we investigated the role of pyomelanin in resistance to oxidative stress and virulence in strains J2315 and K56-2, two epidemic CF isolates belonging to the Burkholderia cenocepacia ET-12 lineage. We previously reported that a single amino acid change from glycine to arginine at residue 378 in homogentisate 1,2-dioxygenase (HmgA) affects the pigment production phenotype: pigmented J2315 has an arginine at position 378, while non-pigmented K56-2 has a glycine at this position. Herein, we performed allelic exchange to generate isogenic non-pigmented and pigmented strains of J2315 and K56-2, respectively, and tested these to determine whether pyomelanin contributes to the protection against oxidative stress in vitro as well as in a respiratory infection in CGD mice in vivo. Our results indicate that the altered pigment phenotype does not significantly impact these strains' ability to resist oxidative stress with H2O2 and NO in vitro and did not change the virulence and infection outcome in CGD mice in vivo suggesting that other factors besides pyomelanin are contributing to the pathophysiology of these strains.IMPORTANCEThe Burkholderia cepacia complex (Bcc) is a group of Gram-negative opportunistic bacteria that are often associated with fatal pulmonary infections in patients with impaired immunity, particularly those with cystic fibrosis and chronic granulomatous disease (CGD). Some Bcc strains are known to naturally produce pyomelanin, a brown melanin-like pigment known for scavenging free radicals and overcoming the host cell oxidative burst. We investigated the role of pyomelanin in Burkholderia cenocepacia strains J2315 (pigmented) and K56-2 (non-pigmented) and performed allelic exchange to generate isogenic non-pigmented and pigmented strains, respectively. Our results indicate that the altered pigment phenotype does not significantly impact these strains' ability to resist H2O2 or NO in vitro and did not alter the outcome of a respiratory infection in CGD mice in vivo. These results suggest that pyomelanin may not always constitute a virulence factor and suggest that other features are contributing to the pathophysiology of these strains.

Colistin, the last resort antibiotic: challenges in the implementation of its routine susceptibility testing

Background: colistin has become a critical antibiotic for lifethreatening multidrug resistance Gram-negative infections, particularly carbapenemase-producing bacteria. Detecting colistin resistance in routine microbiology laboratories is crucial for combating these fatal infections poses a challenge. Especially in developing countries, there is a need for a cost-effective, rapid, and user-friendly diagnostic method. Objective: implementing the various available methods for colistin testing is a significant challenge in resource-limited settings due to logistic difficulties and the need for technical expertise. Materials and Methods: this study shares experiences and insights gained while implementing in-vitro colistin susceptibility testing in a high-load bacteriology laboratory of a tertiary care center in Delhi, India. The following test methods for colistin susceptibility testing were incorporated in the routine antimicrobial susceptibility testing of our laboratory: Colistin Agar Test, Colistin Broth Disk Elution Test, Broth Microdilution susceptibility testing. Results: inconsistent growth patterns were observed in the colistin agar dilution Minimum Inhibitory Concentration (MIC) method, which could be resolved only after the preparation of fresh plates containing that specific concentration of colistin. The contamination issue of plates on use over a few days was addressed by pouring agar containing various concentrations of colistin in cottonplugged glass tubes. In the colistin broth disk elution test, due to the non-availability of screw-capped 10 mL glass tubes, MacCornety bottles (30 mL) were used. Subcultures were performed from the turbid wells to rule out the growth of contaminants when encountering discordant MIC values or skipped wells on the colistin broth microdilution test. Conclusions: despite several technical issues in in-vitro colistin susceptibility testing, we have successfully implemented it in our laboratory. Our experiences can offer guidance to laboratories that are still in the process of implementing it.

Glucans: A Therapeutic Alternative for Sepsis Treatment.

Sepsis treatment is a challenging condition due to its complexity, which involves host inflammatory responses to a severe and potentially fatal infection, associated with organ dysfunction. The aim of this study was to analyze the scientific literature on the immunomodulatory effects of glucans in a murine model of systemic infection induced by cecal ligation and puncture. This study comprises an integrative literature review based on systematic steps, with searches carried out in the PubMed, ScienceDirect, Scopus, Web of Science, and Embase databases. In most studies, the main type of glucan investigated was β-glucan, at 50 mg/kg, and a reduction of inflammatory responses was identified, minimizing the occurrence of tissue damage leading to increased animal survival. Based on the data obtained and discussed in this review, glucans represent a promising biotechnological alternative to modulate the immune response and could potentially be used in the clinical management of septic individuals.

Salmonella pathogenicity island-14 is a critical virulence factor responsible for systemic infection in chickens caused by Salmonella gallinarum

Salmonella enterica serovar Gallinarum (S. gallinarum) is an important host-specific pathogen that causes fowl typhoid, a severe systemic, septicemic, and fatal infection, in chickens. S. gallinarum causes high morbidity and mortality in chickens and poses a significant burden and economic losses to the poultry industry in many developing countries. However, the virulence factors and mechanisms of S. gallinarum-induced systemic infection in chickens remain poorly understood. In this study, we constructed a Salmonella pathogenicity island-14 (SPI-14) mutant strain (mSPI-14) of S. gallinarum and evaluated the pathogenicity of mSPI-14 in the chicken systemic infection model. The mSPI-14 exhibited the same level of bacterial growth and morphological characteristics but significantly reduced resistance to bile acids compared with the wild-type (WT) strain in vitro. The virulence of mSPI-14 was significantly attenuated in the chicken oral infection model in vivo. Chickens infected with WT showed typical clinical symptoms of fowl typhoid, with all birds succumbing to the infection within 6 to 9 days post-inoculation, and substantial increases in bacterial counts and significant pathological changes in the liver and spleen were observed. In contrast, all mSPI-14-infected chickens survived, the bacterial counts in the organs were significantly lower, and no significant pathological changes were observed in the liver and spleen. The expression of interleukin (IL)-1β, IL-12, CXCLi1, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ in the liver of mSPI-14-infected chickens were significantly lower than those in the WT-infected chickens. These results indicate that SPI-14 is a crucial virulence factor in systemic infection of chickens, and avirulent mSPI-14 could be used to develop a new attenuated live vaccine to prevent S. gallinarum infection in chickens.

COVID-19 Associated Mucormycosis and Risk Factors: A Case-Control Study from Turkey

Background: Mucormycosis, a fatal fungal infection, has increased during the COVID-19 pandemic and posed significant challenges for clinicians. Objectives: Our research focused on identifying the clinical traits of patients with COVID-19-associated mucormycosis (CAM), comparing them with a control group, and identifying risk factors for the development of CAM. Methods: Our study was conducted on 39 CAM patients and 78 control patients from September 2020 to October 2022 at a tertiary education center and regional hospital. The control group was selected blindly in a 1:2 ratio among patients who did not develop mucormycosis, were hospitalized due to COVID-19, and were either discharged or deceased. The control group was matched to the case group regarding age and hospitalization date. To test potential risk factors for CAM, we performed a binary logistic regression analysis. The variables included in the multivariate binary logistic regression model were gender, diabetes, cumulative steroid dose (dexamethasone equivalent), duration of steroid treatment, and tocilizumab/anakinra treatment. Results: In our study, 39 patients were diagnosed with CAM. The average age of the patients was 66 ± 11.5 years. Of the patients, 54.7% (n = 64) were male, with a statistically significantly higher proportion of men in the CAM group (74.4% vs. 44.9%, P = 0.003). The diabetes rate was 51.3% (n = 60) among all patients, and it was higher in the CAM group (69.2% vs. 42.3%, P = 0.006). Regarding in-hospital mortality, the rate was higher in the CAM group (56.4% vs. 14.1%, P < 0.001). The median length of stay in the hospital was 37 days for the CAM group and 10 days for the control group (P < 0.001). The cumulative steroid dose was elevated in the CAM group compared to the control group (191 ± 61.4 mg vs. 117 ± 69.8 mg, P < 0.001). The duration of steroid treatment was 16.5 ± 6.2 days in the CAM group, compared to 9.8 ± 4.7 days in the control group (P < 0.001). Among CAM cases, paranasal involvement was the most common (56.4%), followed by rhino-orbital involvement (33.3%). In binary logistic regression analysis, male gender (OR, 3.9; 95% CI, 1.4 – 11.3), diabetes mellitus (OR, 4.4; 95% CI, 1.5 – 12.4), more than ten days of steroid use (OR, 5.5; 95% CI, 1.3 – 22.4), and tocilizumab/anakinra use (OR, 0.23; 95% CI, 0.06 – 0.8) were identified as risk factors for the development of CAM (p values 0.011, 0.005, 0.019, and 0.020, respectively). Conclusions: Male gender, diabetes mellitus, and steroid use for more than ten days were identified as positive risk factors, while tocilizumab/anakinra use was identified as a negative risk factor for the development of CAM.

Enzyme-Linked Immunosorbent Assay Using Henipavirus-Receptor EphrinB2 and Monoclonal Antibodies for Detecting Nipah and Hendra Viruses.

The Nipah virus (NiV) and the Hendra virus (HeV) are highly pathogenic zoonotic diseases that can cause fatal infections in humans and animals. Early detection is critical for the control of NiV and HeV infections. We present the development of two antigen-detection ELISAs (AgELISAs) using the henipavirus-receptor EphrinB2 and monoclonal antibodies (mAbs) to detect NiV and HeV. The NiV AgELISA detected only NiV, whereas the NiV/HeV AgELISA detected both NiV and HeV. The diagnostic specificities of the NiV AgELISA and the NiV/HeV AgELISA were 100% and 97.8%, respectively. Both assays were specific for henipaviruses and showed no cross-reactivity with other viruses. The AgELISAs detected NiV antigen in experimental pig nasal wash samples taken at 4 days post-infection. With the combination of both AgELISAs, NiV can be differentiated from HeV. Complementing other henipavirus detection methods, these two newly developed AgELISAs can rapidly detect NiV and HeV in a large number of samples and are suitable for use in remote areas where other tests are not available.

Current approach to diagnosing and treating necrotizing fasciitis

Introduction: Necrotizing fasciitis is an infrequent yet highly fatal bacterial infection characterized by widespread necrosis of fascia and subcutaneous fat tissue. Though initial symptoms resemble typical infection, necrotizing fasciitis progresses rapidly triggering acute phase response. Individuals with advanced age, chronically ill, immunocompromised, or abusing alcohol are especially susceptible to developing necrotizing fasciitis. In order to reduce mortality, early diagnosis and appropriate treatment are indispensable. Aim of the Study: Aim of this study is through evaluating existing literature to outline the contemporary diagnostic strategies and emerging therapy options for necrotizing fasciitis. Description of the State of Knowledge: Primary diagnostic methods involve clinical evaluation and surgical exploration, complemented by fresh frozen sections for rapid diagnosis and the finger test. Diagnostic imaging incorporates the use of magnetic resonance imaging, computed tomography and ultrasound. The treatment is mostly centered around surgical debridement and antibiotic therapy. Therapies that display potential efficacy include low-dose radiotherapy, hyperbaric oxygen therapy, and the use of intact fish skin grafts for tissue reconstruction after successful treatment. The emerging approach includes therapy targeting systemic inflammatory response syndrome, sepsis induced coagulopathy and critical illness related corticosteroid insufficiency. Conclusions: The management of necrotizing fasciitis primarily depends on traditional methods. Fast identification and proper treatment are pivotal in reducing the mortality rate. Recognizing the significance of addressing the acute phase response in necrotizing fasciitis treatment introduces new possibilities for therapeutic interventions. Further research is vital to evaluate the existing approaches to necrotizing fasciitis management and explore new diagnoistic and therapeutic alternatives.

High-throughput sequencing of Strongyloides stercoralis - a fatal disseminated infection in a dog.

The rhabditid nematode Strongyloides stercoralis is known worldwide as the causative agent of strongyloidiasis in humans. In addition to public health concerns, S. stercoralis also infects dogs, which represent a possible reservoir for potentially zoonotic transmissions. We describe the first confirmed case of fatal disseminated infection in a dog in the Czech Republic. The microscopic and histological results were supported by a complex genotyping approach. Using high-throughput sequencing of the hypervariable region (HVR-IV) of 18S rDNA and Sanger sequencing of the partial cytochrome c oxidase subunit 1 gene (cox1), the potentially zoonotic haplotype/lineage A of S. stercoralis was confirmed, while the solely canine haplotype/lineage B was not found. The development of the disease is mainly associated with immunodeficiency, and in this case, it was triggered by inappropriate treatment, in particular the use of corticosteroids.

Exploring the extrachromosomal plasmid rDNA of Naegleria fowleri AY27 genotype II: A human brain-eating amoeba via high-throughput sequencing

Naegleria fowleri, also known as brain-earing amoeba, causes severe and rapidly fatal CNS infection in humans called primary amebic meningoencephalitis (PAM). The DNA from the N. fowleri clinical isolate was sequenced for circular extrachromosomal ribosomal DNA (CERE - rDNA). The CERE contains 18 S, 5.8 S, and 28 S ribosomal subunits separated by internal transcribed spacers, 5 open reading frames (ORFs), and mostly repeat elements comprising 7268 bp out of 15,786 bp (46%). A wide variety of variations and recombination events were observed. Finally, the ORFs that comprised only 4 hypothetical proteins were modeled and screened against Zinc drug-like compounds. Two compounds [ZINC77564275 (ethyl 2-(((4-isopropyl-4 H-1,2,4-triazol-3-yl) methyl) (methyl)amino) oxazole-4-carboxylate) and ZINC15022129 (5-(2-methoxyphenoxy)-[2,2’-bipyrimidine]-4,6(1 H,5 H)-dione)] were finalized as potential druggable compounds based on ADME toxicity analysis. We propose that the compounds showing the least toxicity would be potential drug candidates after laboratory experimental validation is performed.

IRF7 Exacerbates Candida albicans Infection by Compromising CD209-Mediated Phagocytosis and Autophagy-Mediated Killing in Macrophages.

IFN regulatory factor 7 (IRF7) exerts anti-infective effects by promoting the production of IFNs in various bacterial and viral infections, but its role in highly morbid and fatal Candida albicans infections is unknown. We unexpectedly found that Irf7 gene expression levels were significantly upregulated in tissues or cells after C. albicans infection in humans and mice and that IRF7 actually exacerbates C. albicans infection in mice independent of its classical function in inducing IFNs production. Compared to controls, Irf7-/- mice showed stronger phagocytosis of fungus, upregulation of C-type lectin receptor CD209 expression, and enhanced P53-AMPK-mTOR-mediated autophagic signaling in macrophages after C. albicans infection. The administration of the CD209-neutralizing Ab significantly hindered the phagocytosis of Irf7-/- mouse macrophages, whereas the inhibition of p53 or autophagy impaired the killing function of these macrophages. Thus, IRF7 exacerbates C. albicans infection by compromising the phagocytosis and killing capacity of macrophages via regulating CD209 expression and p53-AMPK-mTOR-mediated autophagy, respectively. This finding reveals a novel function of IRF7 independent of its canonical IFNs production and its unexpected role in enhancing fungal infections, thus providing more specific and effective targets for antifungal therapy.

Comorbidities and final outcome of post COVID-19 associated oral mucormycosis patients: a cross-sectional study

BackgroundMucormycosis is an invasive and possibly fatal fungal infection that frequently affects the mouth. Co-infection in post COVID-19 patients is an emerging concern, owing to their complex nature and higher mortality. Therefore, this study aimed to report the underlying risk factors, clinical characteristics, presentation, and final outcome of COVID-19 associated oral mucormycosis cases.ResultsThis study included 51 patients with oral mucormycosis 28 (55%) males and 23 (45%) females. Twenty-three patients (45%) presented with deep necrotic ulcer, 16 patients (31%) presented with superficial ulcer, and 12 patients (23%) of them presented with fistula. The overall survival rate was 51% (26 patients recovered). On the other hand, mortality rate was 49% (25 patients), of which 68% suffered from uncontrolled diabetes mellitus.ConclusionsThe mortality rate of COVID-19 associated mucormycosis cases with oral manifestations was found to be quite high mostly due to delayed diagnosis and lack of timely treatment. Thus, increasing the awareness of dentists and maxillofacial surgeons is crucial for the early diagnosis of oral mucormycosis which is vital to improve prognosis of this serious infection. This is of high importance in view of the escalating numbers of mucormycosis cases being recorded worldwide.

Monocyte Production of C1q Potentiates CD8+ T-Cell Function Following Respiratory Viral Infection.

Respiratory viral infections remain a leading cause of morbidity and mortality. Using a murine model of human metapneumovirus, we identified recruitment of a C1q-expressing inflammatory monocyte population concomitant with viral clearance by adaptive immune cells. Genetic ablation of C1q led to reduced CD8+ T-cell function. Production of C1q by a myeloid lineage was necessary to enhance CD8+ T-cell function. Activated and dividing CD8+ T cells expressed a C1q receptor, gC1qR. Perturbation of gC1qR signaling led to altered CD8+ T-cell IFN-γ production, metabolic capacity, and cell proliferation. Autopsy specimens from fatal respiratory viral infections in children exhibited diffuse production of C1q by an interstitial population. Humans with severe coronavirus disease (COVID-19) infection also exhibited upregulation of gC1qR on activated and rapidly dividing CD8+ T cells. Collectively, these studies implicate C1q production from monocytes as a critical regulator of CD8+ T-cell function following respiratory viral infection.

Abstract PO1-07-05: Preliminary Evaluation in Breast Cancer of PointCheck: a Noninvasive AI-Based Neutropenia Monitoring Device

Abstract Background Febrile neutropenia (FN) is a common complication that can arise in breast cancer patients receiving myelosuppressive chemotherapy and increases the risk of fatal infections. Despite better preventive strategies there are between 140,000 to 200,000 FN-related hospitalizations yearly. Quick detection and awareness of grade III neutropenia (Absolute Neutrophil Count [ANC] < 1,000/µL) or greater can be critical to managing FN and treatment scheduling and dosing in patients with breast cancer. The current gold standard for neutropenia detection is a peripheral blood collection that requires patients to come into the clinic. PointCheck™ is a novel, noninvasive technology that can monitor for grade III neutropenia or greater and enable prompt detection in the home by acquiring microscopy videos of superficial capillaries through the nailfold skin and analyzing those videos with computer vision AI algorithms. To evaluate the usability, diagnostic performance, and preliminary clinical utility of PointCheck™ on FN detection we conducted a multi-center observational usability and diagnostic study. Methods The present study included an overall cohort of 175 diverse cancer patients, with 70 breast cancer patients. The primary endpoint was to achieve a score over 80.8 on a standardized System Usability Scale (SUS). Secondary endpoint was achievement of diagnostic performance with an area under the curve (AUC) above 0.80. We also performed an exploratory analysis on the device's clinical utility. Eligible participants were introduced to the device, watched a tutorial video and were given a user manual. They used the device autonomously under the supervision of the clinical team. Usability data was collected using the SUS, the scoring system that has a scale between 0-100. PointCheck™ measurements were analyzed using AI algorithms and compared to same-day Complete Blood Counts (CBCs) collected within 90 minutes of the measurement to assess accuracy in classifying patients as grade III neutropenic or greater (< 1,000/µL), or non-grade III neutropenic (≥1,000/µL). In the exploratory clinical utility analysis, clinicians provided narrative insights by completing a Likert survey assessing how PointCheck™ would support or impact their clinical decisions. Results 81.4% of breast cancer patients scored above 80.8 on the SUS scale across all sites, with a mean SUS score of 87.7 (SD=13.2). Furthermore, the AI-based PointCheck™ classifier accurately discriminated neutropenia patients with an AUC = 0.90 in the overall cohort. In the exploratory analysis, a majority of clinicians (65.2%) agreed that the device could have helped better evaluate their patients (Table 1). Whereas almost 3 out of 4 clinicians (72.4%), described that the use of PointCheck™ could have avoided preparing a chemotherapy infusion if the patient was not ready due to grade III neutropenia or greater. Conclusions The present study showed that there is a high perception of usability of PointCheck™, indicating an above average user experience and falling within the top 10% of systems. Furthermore, we found that PointCheck™ can accurately detect grade III neutropenia or greater in a large cohort of patients including breast cancer patients. Application of PointCheck™ to clinical practice as a monitoring system can contribute to the early detection of FN and antineoplastic therapy management. Table 1: Clinical Utility Survey Results Citation Format: Juan Garcia-Mosquera, Manuel Alva, Toshiaki Iwase, Ganimete Lamaj, Pablo Tolosa, Rodrigo Sánchez-Bayona, José Pelaez-Escobar, Alberto Pablo-Trinidad, Ryan Benasutti, Aurélien Bourquard, Ian Butterworth, Alvaro Sánchez-Ferro, Carlos Castro-González, José Manuel Pérez-García, Alejandro Martínez-Bueno, Eva Ciruelos, Azadeh Nasrazadani. Preliminary Evaluation in Breast Cancer of PointCheck: a Noninvasive AI-Based Neutropenia Monitoring Device [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO1-07-05.

Clinical Characteristics and Prognoses of Mucormycosis in Four Children.

Mucormycosis is a fatal invasive fungal infection that commonly affects immunocompromised children. The aim of our study was to investigate the clinical manifestations, treatments, and prognosis of pediatric patients with mucormycosis. We conducted a retrospective search in Shenzhen Children's Hospital from July 2013 to July 2023 for all patients with mucormycosis. The clinical manifestation, pathogen detection, radiology, treatments, and prognosis were analyzed. Four cases were identified. Underlying conditions included acute myeloid leukemia with myeloid sarcoma (n = 1), thalassemia (post-allogeneic hematopoietic stem cell transplantation; n = 1), systemic lupus erythematosus (n = 1), and bilateral nephroblastoma (post-bilateral nephrectomy; n = 1). Two patients were disseminated mucormycosis, one case was pulmonary mucormycosis, and one case was cerebral mucormycosis. Fever, cough, and dyspnea were the main clinical symptoms of pulmonary mucormycosis, headache was the main clinical symptom of cerebral mucormycosis. Lung CT findings included consolidation, multiple nodules, halo sign, air crescent sign, and pleural effusion. The contrast-enhanced CT showed pulmonary artery and pulmonary vein occlusions in two patients and pseudoaneurysm in two patients. Amphotericin B formulations were administered as first-line therapy in all cases; in three cases, Triazole was administered in combination with amphotericin B. Mucormycosis is a life-threatening disease involving multiple systems. Aorta pseudoaneurysm is a rare and fatal complication, enhanced CT can assist in diagnosis. Early diagnosis and appropriate therapeutic strategies are needed.

Reversible Cytotoxic Edema in Patients with COVID-19 Associated Encephalitis Presenting Status Epilepticus

Coronavirus disease 2019 (COVID-19) is a serious infectious disease with multisystem alteration including neurological complications. COVID-19 associated encephalitis is a potentially fatal viral infection of the brain. Diffusion-weighted images (DWI) are a useful evaluation modality of cytotoxic edema in patients with encephalitis. We report on the reversible DWI change in a patient with COVID-19 associated encephalitis, who had diffused cytotoxic edema in the affected temporal lobe and accompanied status epilepticus. Sequential imaging showed that the cytotoxic edema in DWI confirmed recovery to normal after 12 days in the presence of cortical edema in fluid-attenuated inversion recovery image after aggressive neurocritical management. Thus, prompt, proper management is indispensable during the acute period and DWI may be a valuable tool for reflecting the clinical course of COVID-19 associated encephalitis.

Abstract 112: Endothelial Mixed Lineage Leukemia 1 (kmt2a/MLL1) As A Central Mediator Of Immunothrombosis During SARS-CoV-2 Infection

Background: Endotheliitis in severe SARS-CoV-2 (CoV2) results in increased cellular adhesion molecules (CAMs) and procoagulant molecules (PCMs). Cell surface CAMs increase endothelial cell (EC) interactions with circulating immune cells (IC). Increased EC-IC interactions and a procoagulant EC phenotype augment immunothrombotic risk during CoV2 infection. Prophylactic anticoagulants during CoV2 infection are associated with unacceptably high bleeding risk. New targets to treat endotheliitis are required. Hypothesis: We hypothesize chromatin modifying enzymes (CME)s associated with inflammatory NF-kB signaling drive transcriptional changes in endothelial cells (ECs) leading to a prothrombotic and procoagulant phenotype. Aims: Identify potential CMEs that drive CoV-2 endotheliitis. Methods: Public human single cell RNA sequencing data evaluated the pulmonary endothelial epigenetic transcriptome (PMID34876692). Murine venous ECs (mVECs), EOMA cells, and human venous ECs (HUVECs) were treated with murine beta-coronavirus (MHV-A59) or CoV2 spike protein (S1), respectively. Pharmacologic and siRNA inhibition were used to accomplish MLL1 silencing. Results: Humans with fatal CoV2 infection demonstrated marked increase in the KMT2/MLL class of histone methyltransferases in pulmonary ECs. mVEC infection significantly increased transcription of CME kmt2a, CAM esel, and PCMs plau and f3. Significant enrichment of H3K4 methylation and MLL1 are seen on promotors of identified genes. Inhibition of MLL1 reduced CAM and PCM transcripts. Functional EC-IC adhesion testing revealed significant decreases in monocyte adhesion with MLL1 inhibition. Conclusion: Kmt2a/MLL1 positively regulates CoV-2 induction of CAMs and PCMs, endothelial inflammation via positive regulation of CAM gene transcription, and resultant EC-IC interactions. MLL1 may represent a novel therapeutic target to ameliorate immunothrombotic risk during CoV-2 infection.

A novel indirect ELISA for serodiagnosis of mucormycosis using antigens from Rhizopus arrhizus.

Due to a delay in diagnosis by conventional techniques and high mortality, the development of a standardised and rapid non-culture-based technique is an unmet need in pulmonary, gastrointestinal, and disseminated forms of mucormycosis. Though limited studies have been conducted for molecular diagnosis, there are no established serologic tests for this highly fatal infection. To develop and evaluate an indirect in-house enzyme-linked immunosorbent assay (ELISA) utilising antigens of Rhizopus arrhizus for detecting anti-Rhizopus antibodies (IgG and IgM) in sera of patients with mucormycosis. We extracted both secretory and mycelial Rhizopus antigens using standardised protocols. Bradford assay was used for protein quantification. We then standardised an indirect ELISA using R. arrhizus mycelial and secretory antigens (10.0 μg/mL in bicarbonate buffer pH 9.2) for detecting anti-Rhizopus IgG and IgM antibodies in patient sera. We included patients with mucormycosis, other fungal infections, and healthy controls. Antibody index value (E-value) was calculated for each patient sample. Asparagine broth culture filtrate utilising 85% ammonium sulphate salt fractionation and mycelial homogenate grown in yeast extract peptone dextrose (YPD) broth precipitated with trichloroacetic acid (TCA) yielded a large amount of good-quality protein for the assay. We included 55 patients with mucormycosis (rhino-orbito-cerebral mucormycosis [ROCM, n = 39], pulmonary [n = 15], gastrointestinal [n = 1]), 24 with other fungal infections (probable aspergillosis [n = 14], candidiasis [n = 10]), and healthy controls (n = 16). The sensitivity of the antibody test for diagnosing mucormycosis ranged from 83.6-92.7% for IgG and 72.7-87.3% for IgM, with a specificity of 91.7-92.5% for IgG and 80-82.5% for IgM. The sera from patients with other fungal infections and healthy individuals did not show significant cross-reactivity. The detection of anti-Rhizopus IgG antibody performed significantly better in comparison to IgM-based ELISA for diagnosing both ROCM (sensitivity of 84.6% vs. 69.2%) and pulmonary cases (86.6% vs. 80.0%). More extensive studies are required to confirm our findings.