A Multiplex PCR and DNA-Sequencing Workflow on Serum for the Diagnosis and Species Identification for Invasive Aspergillosis and Mucormycosis.

There has been significant increase in the use of molecular tools for the diagnosis of invasive aspergillosis (IA) and mucormycosis. However, their range of detection may be too limited as species diversity and coinfections are increasing. Here, we aimed to evaluate a molecular workflow based on a new multiplex PCR assay detecting the whole Aspergillus genus and the Mucorales order followed by a species-specific PCR or a DNA-sequencing approach for IA and/or mucormycosis diagnosis and species identification on serum. Performances of the MycoGENIE Aspergillus spp./Mucorales spp. duplex PCR kit were analyzed on a broad range of fungal strains and on sera from high-risk patients prospectively over a 12-month period. The kit allowed the detection of nine Aspergillus species and 10 Mucorales (eight genera) strains assessed. No cross-reactions between the two targets were observed. Sera from 744 patients were prospectively analyzed, including 35 IA, 16 mucormycosis, and four coinfections. Sensitivity varies from 85.7% (18/21) in probable/proven IA to 28.6% (4/14) in COVID-19-associated pulmonary aspergillosis. PCR-positive samples corresponded to 21 A. fumigatus, one A. flavus, and one A. nidulans infections. All the disseminated mucormycosis were positive in serum (14/14), including the four Aspergillus coinfections, but sensitivity fell to 33.3% (2/6) in localized forms. DNA sequencing allowed Mucorales identification in serum in 15 patients. Remarkably, the most frequent species identified was Rhizomucor pusillus (eight cases), whereas it is barely found in fungal culture. This molecular workflow is a promising approach to improve IA and mucormycosis diagnosis and epidemiology.

COVID-19 vaccine immunogenicity among CD19 receptor T-cell (CAR-T) therapy

Background. Patients receiving CAR-T therapy may have impaired humoral responses to SARS-CoV-2 vaccinations due to their high net state of immunosuppression associated with the underlying disease, prior lines of therapy and CAR-T treatment associated hypogammaglobinemia. Comprehensive data on vaccine immunogenicity in this patient population are currently lacking. Methods. A single-center retrospective study of adults receiving CD19 CAR-T therapy for non-Hodgkin's lymphoma was conducted between 3/27/2018 - 8/31/ 2021. Patients received at least two doses of COVID-19 vaccinations with BNT162b2 (Pfizer, BioNTech), mRNA-1273 (Moderna), or 1 dose of Ad26.COV2.S (Janssen) and had SARS-CoV-2 anti-spike (S) levels measured at least one month after the last vaccine dose. We excluded patients who received COVID-19 monoclonal antibody therapy or immunoglobulin within 3 months of the index anti-S titer. Patients were followed from the time of the first COVID-19 vaccines through their index anti-S antibody result. Patients were censored on the first day of any additional antineoplastic therapy after disease relapse. Our primary endpoint was the percentage of patients who develop a positive anti-S response (assessed by anti-S assay cutoff of >0.8 U/mL, Roche assay). Results. Twenty-five patients met eligibility. Median age was 65 years (range 41 - 78), and majority of patients were male (72%). The number of patients with a positive antibody response was 12 (48%). Median number of vaccines received was 3. 18 patients (72%) received Pfizer vaccines, 4 patients (16%) received Moderna, 2 patients (8%) received Moderna and Pfizer, and 1 patient (4%) received Janssen and Pfizer. Median anti-S titers among patients with a positive response was 111 U/mL (range 2.44 - 12500). Two patients (8%) had COVID-19, both with negative anti-S responses. Conclusion. Our analysis shows that only 48% of patients who received CAR-T therapy developed a positive antibody response after at least two COVID-19 vaccine doses, with a low median titer among responders. This patient population is at higher risk for developing severe COVID-19 disease and likely remains vulnerable even after vaccination. Alternative approaches are needed to prevent COVID-19 and mitigate disease severity in patients undergoing CAR-T.

Cross-sectional study to determine the prevalence of telogen effluvium among patients experiencing COVID-19 infection or vaccination in Saudi Arabia and Arabic countries

Background: Telogen effluvium (TE) after COVID-19 infection or vaccination is a common sequelae in dermatology clinics. Objective(s): to study the prevalence of telogen effluvium in COVID-19 patients and its association with COVID vaccine. Method(s): Cross-sectional study via online questionnaire in Saudi Arabia and other Arabic countries. Result(s): Prevalence of hair loss among COVID-19 patients is ~85% with 45.9% meeting the criteria of TE. Majority of those with high fever associated with COVID-19 developed TE (87.5%). 100% of hospitalized patients exhibited TE with 58% having diffuse hair loss for less than 6 months (acute TE) and 32% for more than 6 months (chronic TE). 63.2% of our participants had hair loss after COVID vaccination regardless of vaccine type with the majority experiencing it after the first dose (55.8%). Limitation(s): The majority of the participants were female. Other factors associated with hair loss were taken into consideration. Conclusion(s): COVID-19 infection and its vaccines carry a high risk for development of telogen effluvium. Copyright © 2022 Ubiquity Press. All rights reserved.

A proposition of a latent needs identifying method based on an experiment of working prototype-based interview

The purpose of this research is to verify the method in the elicitation of latent needs from consumer needs by conducting the working prototype-based interview and collecting raw data which is responses from the consumer. Latent needs are those that many consumers recognize as important in a final product but do not or cannot articulate in advance. The challenge in identifying latent needs is finding the method to elicit from consumers the needs which are not addressed by any inventors yet in the present market but would delight consumers if delivered tomorrow. After conducting a consumer feedback questionnaire via the internet, a working prototype was created. The working prototype then was used as material to prepare presentation slides. The first presentation slides were focused on the background problems and ideas for the solutions while the second presentation slides provided consumers with a prototype and story of the product that was believed would be one of the solutions to the problems. Based on the results, latent needs interpreted from interviewees’ responses and the categories of the needs obtained from the Prototype-based interviews are more than from the Problem-based interview. The latent needs that we were able to obtain from this research were for example, “The device is able to detect small changes in a child while changing a diaper” and “The device is able to detect small changes in a child while watching he/she sleeping” which could lead into the prevention of unwanted incident such as sudden infant death syndrome (SIDS). This supports our assumption that showing working prototype based materials with story descriptions can be effective in uncovering potential latent needs. However, due to the COVID-19 pandemic, we were unable to give the interviewees chances to touch and look closely at the working prototype therefore latent needs possibly gained from this experience are still uncovered. Although there are still limitations in our findings, the method that we proposed is able to support discovering latent needs in future. © 2022 The Japan Society of Mechanical.

The “New Caledonia COVID-19 Paradox”: Dramatic indirect impact of the pandemic on organ donation and transplantation in a non-prevalence country

Introduction With 270,000 inhabitants, New Caledonia has a very high prevalence of End-stage Kidney Disease (ESKD), with the fourth-highest globally. New Caledonia started a deceased and live-donor kidney transplantation program five years ago. On March 18th, 2020, the New Caledonian government halted all flights because of potential risk of COVID circulation. The consequences in term of ESKD patient management was significant. Description The purpose was to evaluate the impact of the COVID epidemic in 2020 on the rate of kidney transplantation in an isolated country free of local COVID cases over the period of one year. Methods Number of Covid tests performed in the general population, number of positive tests and number of transplantation performed over the year 2020 were collected. Results An extensive screening program was implemented with an average COVID-19 RT-PCR test of 14,800 per million. Data showed that COVID-19 didn’t circulate among the population. Only a small number of imported cases were diagnosed in the context of a comprehensive screening policy and strict isolation rules. This government policy was very effective in protecting the country from the direct consequences of COVID-19, but it dramatically impacted kidney transplantation. No patient could be transplanted through the Australian and New Zealand kidney paired exchange program (ANZKX). Only 11 kidney transplants were performed in 2020 compared to an average of 23 transplants for the four preceding years, representing a nearly 50% drop in transplant activities (Fig. 1). Conclusion Halting elective surgeries was deleterious on the newly-established living-donor transplant program established less than one year prior in almost complete autonomy. The reduced mobility of surgeons from expert centers in France and Australia was an essential factor. The 2020 COVID-19 pandemic protective closure measures nipped this country's local kidney transplant program in the bud.

COVID-19 Patient' s Background Frequency on EEG Predictive of Hospital Discharge Disposition

Objective: To determine whether background frequency on encephalogram (EEG) was associated with hospital discharge disposition in COVID-19 patients. Background: Neurological complications, including encephalopathy, have been reported up to 80% of hospitalized COVID-19 patients. It is unclear how encephalopathy during acute illness is related to clinical outcomes in COVID-19 patients. Design/Methods: Subjects were drawn from hospitalized patients who received an EEG at single tertiary center between March 2020 and February 2021. Included were adult subjects with confirmed COVID-19 during hospitalization who were not in the intensive care unit at the time of EEG evaluation. Clinical outcome measured was disposition upon discharge, graded as variables: 0-home, 1-acute rehabilitation, 2-subacute rehabilitation, 3-long term acute care hospital, and 4-death. Background frequency on EEG was used as a measure of encephalopathy severity and was assessed in occipital channels. EEG and medical records were reviewed retrospectively. We used two-independent t test for univariate analysis. Significant variables were then analyzed with multivariate linear regression in SPSS. Study was approved by the University of Chicago IRB. Results: A total of 50 subjects were included. Reasons for EEG were concern for seizure (n=14, 28.0%), altered mental status (n=33, 66.0%), and others (n=3, 6.0%). According to WHO COVID-19 severity scale, on admission 37 (74%) patients had ambulatory disease, 11 (22%) had moderate disease, and 2 (4.0%) severe disease. Thirteen (26.0%) patients received COVID-19 treatment, 4 (8.0%) with steroid. Sodium upon admission, history of epilepsy, cefepime use, intubation, and EEG background frequency were all significant predictors of disposition with univariate analysis (p <0.05). Multivariate analysis showed only intubation (B 1.347, p=0.03) and background frequency on EEG (B -0.282, p <0.001) as independent predictors of disposition. Conclusions: EEG background frequency was a predictor of discharge disposition in COVID-19 patients, and patient with low background frequencies were less likely to be discharged home.

L'association tocilizumab et dexaméthasone au cours du COVID-19 sévère est-elle associée à un risque de surinfection ?

Introduction La prise en charge des patients atteints de COVID-19 a changé depuis le début de la pandémie. Le tocilizumab est recommandé en association avec la dexaméthasone en cas de forme inflammatoire sévère. Néanmoins, les données concernant le risque de surinfection de cette association sont encore rares et discordantes, notamment en réanimation : certaines suggèrent un risque accru de surinfection chez ces patients. Matériels et méthodes Nous avons mené une étude rétrospective monocentrique afin de décrire le risque de surinfections bactériennes et fongiques associées au COVID-19 sévère en réanimation chez des patients traités par dexaméthasone avec ou sans tocilizumab. Résultats Entre mars 2020 et août 2021, 246 patients (72 % hommes) d'âge médian 61 ans ont été inclus : 64 (26 %) présentaient une immunodépression et 57 (23 %) n'avaient aucune comorbidité. Un SDRA était présent à l'admission chez 226 (92 %) patients et 56 (23 %) ont nécessité une ventilation invasive. Durant l'étude, 150 (61 %) patients ont reçu de la dexaméthasone et du tocilizumab (groupe 1) et 96 (39 %) patients ont reçu de la dexaméthasone sans tocilizumab (groupe 2). Les scores médians IGS2 et SOFA étaient de 23 et 3, respectivement, sans différence entre les groupes 1 et 2. Au cours du suivi, 59 patients ont développé 66 surinfections : 34 (23 %) patients dans le groupe 1 et 25 (26 %) patients dans le groupe 2 (p=0,32). La surinfection était respiratoire pour 39 patients, documentée par un bacille Gram négatif dans 19 cas (45 %). Ces surinfections sont survenues dans un délai médian de 10 jours après l'admission. Parmi ces 66 surinfections, nous avons documenté 11 aspergilloses pulmonaires dans un délai médian de 9 jours après l'admission. Huit patients avaient une bactériémie et six patients une infection urinaire. Enfin, un patient a développé une pneumocystose et un autre une infection à Clostridioides difficile. En analyse multivariée, les facteurs associés à un sur risque de surinfection étaient la présence d'une hémopathie (OR=2,47), le recours à la ventilation invasive (OR, 3,74) et l'IGS2 à l'admission (OR, 1,03). L'administration de tocilizumab, en plus de la dexaméthasone, n'était pas associée à un sur risque de surinfection (OR=0,61). Conclusion Notre étude montre que la co prescription de tocilizumab et de dexaméthasone en réanimation pour COVID-19 sévère, n'était pas associée à un risque accru de surinfections par rapport à l'administration de dexaméthasone seule. Aucun lien d'intérêt

Immunogenicity and Reactogenicity of COVID-19 mRNA Vaccines in Allogeneic Stem Cell Transplant Recipients

Background. Allogeneic stem cell transplant (SCT) recipients are at an increased risk of poor outcomes from COVID-19. While the mRNA-1273 (Moderna) and BNT162b2 (Pfizer) COVID-19 mRNA vaccines are highly immunogenic in the general population, the immune response in SCT recipients is poorly understood. We characterized the immunogenicity and reactogenicity of COVID-19 mRNA vaccines in a cohort of SCT patients. Methods. We performed a prospective cohort study of 16 allogeneic SCT patients and 23 healthy controls. Blood samples for both cohorts were collected prior to first vaccination (baseline), at the time of second vaccination, and approximately 28 days post-second vaccination. Anti-Spike (S), anti-S1, anti-receptor binding domain (RBD), and anti-Nucleocapsid (N) IgG levels were measured quantitatively from plasma using a multiplexed single molecule array (Simoa) immunoassay. Reactogenicity was captured for the SCT cohort via a self-reported post-vaccination diary for 7 days after each dose. Results. Demographics and SCT recipients' characteristics are shown in Table 1. In the SCT cohort, we observed a significantly lower anti-S (p< 0.0001), S1 (p< 0.0001), and RBD (p< 0.0001) IgG responses as compared to healthy controls, both at the time of dose 2 and 28 days post-vaccine series (Fig 1). Overall, 62.5% of SCT recipients were responders after vaccine series completion, as compared to 100% of healthy controls (Fig 2). While no patients had a reported history of COVID-19 diagnosis, 2 patients in the SCT cohort had elevated anti-S IgG levels and 1 showed elevated anti-N at baseline. 10/16 participants in the SCT cohort completed at least one post-vaccination diary. Local and systemic reactions were reported by 67% and 22% of participants, respectively, after dose 1, and 63% and 50% after dose 2 (Figure 3). All reported events were mild. Anti-Spike (A), anti-S1 (B), anti-RBD (C), and anti-nucleocapsid (D) IgG titers were measured at baseline, time of second dose, and approximately 28 days after second vaccination. IgG levels were measured quantitatively using multiplexed single molecule array (Simoa) immunoassays, and are reported as Normalized Average Enzymes per Bead (AEB). Allogeneic stem cell transplant recipients (mauve) showed significantly lower anti-S, S1, and RBD IgG responses as compared to healthy controls (mint). Low titers of anti-N IgG demonstrates no history of COVID-19 natural infection during the course of the study. 10 allogeneic stem cell transplant recipients completed at least one diary for 7 days after vaccination. Reactions after dose 1 are shown in light blue, and reactions after dose 2 are shown in dark blue. Local reactions (A) were reported by 67% (6/9) of participants after dose 1, and 63% (5/8) after dose 2. Systemic reactions (B) were reported by 22% (2/9) of participants after dose 1, and 50% (4/8) after dose 2. All reported events were mild (Grade 1). Conclusion. Among SCT recipients, mRNA COVID-19 vaccines were well-tolerated but less immunogenic than in healthy controls. Further study is warranted to better understand heterogeneous characteristics that may affect the immune response in order to optimize COVID-19 vaccination strategies for SCT recipients. Figure 2: Response Rate to COVID-19 Vaccination An internally validated threshold for responders was established using pre-pandemic sera from healthy adults. A positive antibody response was was defined as individuals with anti-Spike IgG levels above the 1.07 Normalized AEB threshold.

Immunogenicity of COVID-19 mRNA Vaccines in Patients with Lymphoid Malignancies

Background. Patients with lymphoid malignancies are at high risk of severe COVID-19 disease and were not included in the phase 3 mRNA vaccine trials. Many patients with lymphoid malignancies receive immunosuppressive therapies, including B-cell depleting agents, that may negatively impact humoral response to vaccination. Methods. We recruited patients with lymphoid malignancies and healthy participants who planned to receive two doses of SARS-CoV-2 mRNA vaccine (BNT162b2 or mRNA-1273). Blood was drawn at baseline, prior to second dose of vaccine, and 28 days after last vaccination. Disease characteristics and therapies were extracted from patients' electronic medical record. An ultrasensitive, single molecule array (Simoa) assay detected anti-Spike (S), anti-S1, anti-receptor binding domain (RBD), and anti-Nucleocapsid (N) IgG from plasma at each timepoint. Results. 23 healthy participants and 37 patients with lymphoid malignancies were enrolled (Table 1). Low titers of anti-N (Fig 1A) demonstrate no prior exposure or acquisition of COVID-19 before vaccination or during the study. 37.8% of the lymphoid malignancy cohort responded to the vaccine, using an internally validated AEB cutoff of 1.07. A significantly higher magnitude of anti-S (p< 0.0001), anti-S1 (p< 0.0001) and anti-RBD (p< 0.0001) are present in the healthy as compared to lymphoid malignancy cohort at the second dose and day 28 post-series (Fig 1B, Fig 1C and Fig 1D). Anti-S IgG titers were compared between the healthy cohort, treatment naI&Die;ve, and treatment experienced groups (Fig 2). The treatment naI&Die;ve cohort had high titers by series completion which were not significantly different from the healthy cohort (p=0.2259), although the treatment experienced group had significantly decreased titers (p< 0.0001). Of the 20 patients who had received CD20 therapy, there was no clear correlation of anti-S IgG response with time from CD20 therapy, although most patients who received CD20 therapies within 12 months from the vaccine had no response (Figure 3). Conclusion. The vaccine-induced immune response was poor among treatment-experienced patients with lymphoid malignancies, especially among those who received CD20 therapies within 12 months.