Microclimate factors related to dengue virus burden clusters in two endemic towns of Mexico.

In dengue-endemic areas, transmission control is limited by the difficulty of achieving sufficient coverage and sustainability of interventions. To maximize the effectiveness of interventions, areas with higher transmission could be identified and prioritized. The aim was to identify burden clusters of Dengue virus (DENV) infection and evaluate their association with microclimatic factors in two endemic towns from southern Mexico. Information from a prospective population cohort study (2·5 years of follow-up) was used, microclimatic variables were calculated from satellite information, and a cross-sectional design was conducted to evaluate the relationship between the outcome and microclimatic variables in the five surveys. Spatial clustering was observed in specific geographic areas at different periods. Both, land surface temperature (aPR 0·945; IC95% 0·895-0·996) and soil humidity (aPR 3·018; IC95% 1·013-8·994), were independently associated with DENV burden clusters. These findings can help health authorities design focused dengue surveillance and control activities in dengue endemic areas.

Environmental and socio-economic determinants of the occurrence of malaria clusters in Colombia.

This study identifies the environmental and socio-economic determinants of clusters of high malaria incidence in Colombia during the period of 2008-2019. The malaria cases were obtained from the National System of Surveillance in Public Health, with 798,897 cases reported in the 986 Colombian municipalities evaluated during the study period. Spatial autocorrelation of incidence was examined with global and local indices. Clusters were identified in the Amazon, Pacific, and Uraba-Bajo Cauca-Alto Sinú regions. The factors associated with a municipality belonging to a high-incidence cluster were identified using a logistic regression model with mixed effects and showed a positive association for the variables (forest coverage and minimum multi-year average rainfall). An inverse relationship was observed for aqueduct coverage and the odds of belonging to a cluster. A 1% increase in forest coverage was associated with a 4.2% increase in the odds of belonging to a malaria cluster. The association with minimum multi-year average rainfall was positive (OR = 1.0011; 95% CI 1.0005-1.0027). A 1% increase in aqueduct coverage was associated with a 4.3% decrease in the odds of belonging to malaria cluster. The identification of malaria cluster determinants in Colombia could help guide surveillance and disease control policies.

Study of efficacy and longevity of immune response to third and fourth doses of COVID-19 vaccines in patients with cancer: A single arm clinical trial.

Background: Cancer patients show increased morbidity with COVID-19 and need effective immunization strategies. Many healthcare regulatory agencies recommend administering 'booster' doses of COVID-19 vaccines beyond the standard two-dose series, for this group of patients. Therefore, studying the efficacy of these additional vaccine doses against SARS-CoV-2 and variants of concern is of utmost importance in this immunocompromised patient population. Methods: We conducted a prospective single arm clinical trial enrolling patients with cancer that had received two doses of mRNA or one dose of AD26.CoV2.S vaccine and administered a third dose of mRNA vaccine. We further enrolled patients that had no or low responses to three mRNA COVID vaccines and assessed the efficacy of a fourth dose of mRNA vaccine. Efficacy was assessed by changes in anti-spike antibody, T-cell activity, and neutralization activity, which were again assessed at baseline and 4 weeks. Results: We demonstrate that a third dose of COVID-19 vaccine leads to seroconversion in 57% of patients that were seronegative after primary vaccination series. The immune response is durable as assessed by anti-SARS-CoV-2 (anti-S) antibody titers, T-cell activity, and neutralization activity against wild-type (WT) SARS-CoV2 and BA1.1.529 at 6 months of follow-up. A subset of severely immunocompromised hematologic malignancy patients that were unable to mount an adequate immune response (titer <1000 AU/mL) after the third dose and were treated with a fourth dose in a prospective clinical trial which led to adequate immune boost in 67% of patients. Low baseline IgM levels and CD19 counts were associated with inadequate seroconversion. Booster doses induced limited neutralization activity against the Omicron variant. Conclusions: These results indicate that third dose of COVID vaccine induces durable immunity in cancer patients and an additional dose can further stimulate immunity in a subset of patients with inadequate response. Funding: Leukemia Lymphoma Society, National Cancer Institute. Clinical trial number: NCT05016622.

People with cancer have a higher risk of death or severe complications from COVID-19. As a result, vaccinating cancer patients against COVID-19 is critical. But patients with cancer, particularly blood or lymphatic system cancers, are less likely to develop protective immunity after COVID-19 vaccination. Immune suppression caused by cancer or cancer therapies may explain the poor vaccine response. Booster doses of the vaccine may improve the vaccine response in patients with cancer. But limited information is available about how well booster doses protect patients with cancer against COVID-19. Thakkar et al. show that a third dose of a COVID-19 vaccine can induce a protective immune response in half of the patients with cancer with no immunity after the first two doses. In the experiments, Thakkar et al. tracked the immune reaction to COVID-19 booster shots in 106 cancer patients. A third booster dose protected patients for up to four to six months and reduced breakthrough infection rates to low levels. Eighteen patients with blood cancers and severe immune suppression had an inadequate immune response after three doses of the vaccine; a fourth dose boosted the immune response for two-thirds of them, which for some included neutralization of variants such as Omicron. The experiments show that booster doses can increase COVID-19 vaccine protection for patients with cancer, even those who do not respond to the initial vaccine series. Thakkar et al. also show that pre-vaccine levels of two molecules linked to the immune system, (immunoglobin M and the CD19 antigen) predicted the patients' vaccine response, which might help physicians identify which individuals would benefit from booster doses.

Habilidades de comprensión y producción narrativa en niños montevideanos de 5 años. Una comparación por nivel socioeconómico / Narrative abilities in a group of Uruguayan preschoolers from different socioeconomic backgrounds

Resumen Este estudio caracteriza y compara las habilidades narrativas y el vocabulario receptivo de 54 preescolares montevideanos de nivel socioeconómico medio (NSM) y bajo (NSB). Los niños realizaron tareas de vocabulario receptivo, producción narrativa (en dos condiciones: manipulando títeres al contar la historia y sin manipularlos), y comprensión narrativa (recuento oral y preguntas posteriores a la reconstrucción). Las narrativas fueron transcritas, codificadas y analizadas en términos de super y macroestructura siguiendo la gramática de historias de Stein y Glenn (1979) (coherencia) y de microestructura (productividad y complejidad). Los niños de ambos grupos tuvieron un desempeño acorde a su edad en comprensión y producción narrativa, a pesar de que el grupo de NSM tuvo un mejor desempeño en vocabulario receptivo. Hubo diferencias en comprensión (cantidad de episodios recuperados y en la respuesta a preguntas) que favorecieron al NSM, pero no en la cantidad de categorías recuperadas ni en la extensión y complejidad del recuento. Para producción narrativa no hubo diferencias en secuencia narrativa, ni en la cantidad de episodios completos en ninguna de las condiciones, pero sí en cuanto a la complejidad sintáctica en la condición sin títeres. Los resultados muestran una relación compleja entre vocabulario y habilidades narrativas, dado que diferencias importantes en vocabulario no se reflejaron de manera homogénea en el desempeño narrativo. Estos resultados aportan a los debates actuales sobre el papel del vocabulario en el desarrollo de habilidades narrativas, así como a pensar la validez ecológica de las evaluaciones en el desarrollo cognitivo y lingüístico.

Abstract Narrative abilities are an important part of communication, academic success, and healthy relationships. These abilities involve complex language and cognitive skills, such as precise vocabulary, control of the coherence markers, relations of cause-effect, and planning. They are also relevant during the elementary school years and interact with the socio-emotional skills necessary to understand different points of view. Oral narrative production develops dramatically from 3 to 5 years of age and is a key factor in a child's ability to communicate about the world. During this period narratives are a product of increasing linguistic sophistication over the preschool period and there is a complex relationship between early narratives and language proficiency. So far, most research about this topic has been pursued in populations other than Latin American preschoolers. At the same time, a considerably lesser number of studies about narrative abilities development have been carried out comparing typically developing children from different socioeconomic backgrounds. To our knowledge, there are no studies in Uruguay that assess the narrative abilities development in typically developing preschool children who grow up in vulnerable contexts. It is therefore of the utmost relevance to produce empirical evidence for this population. For these reasons, this study aimed at characterizing and comparing narrative abilities and receptive vocabulary in a group of a total of 54 Uruguayan preschoolers from different socioeconomic backgrounds (middle and low socioeconomic status, SES). Children were assessed in receptive vocabulary and narrative abilities across two task conditions: (1) narrative comprehension through a story retelling task which included some final questions about the story; (2) narrative production elicited from a set of thematically related puppets. The examiner gives a child a puppet set and asks him/her to elaborate a narrative using them. After that, puppets are removed and the child is asked to retell the story without puppets. Children's oral productions were video-taped and then transcribed and categorized using the ELAN software (Max Planck Institute for Psycholinguistics, 2019). The verbal information was categorized according to Stein and Glenn's (1979) story grammar and considering the microstructural aspects (productivity and complexity). Analyses showed that both groups performed according to their stage of development in both narrative task conditions. With regards to the narrative comprehension task, no differences in the number of recovered categories, extension, or narrative complexity were observed. Concerning the narrative production task, there were no differences between the groups in narrative sequences, nor in the number of completed episodes in conditions neither with puppets nor without puppets. Results showed that children who grow up in poverty perform more poorly than their peers from middle-income families in receptive vocabulary, in the number of recovered episodes, ask-answer items of narrative comprehension tasks. Moreover, concerning the microstructural parameters analyses showed that children from middle socioeconomic backgrounds scored better in syntactic complexity in the without puppets condition. No differences between the groups were observed in syntactic complexity in the puppets condition. Taken together these results indicate a complex link between vocabulary and narrative abilities. It is worth noting that important differences in vocabulary did not reflect in the children's narrative performance. These findings are also relevant as a contribution to an ongoing debate about the role of vocabulary in the development of narrative abilities. Furthermore, these results could inform the discussion about the ecological validity of the test of cognitive and linguistic development. Finally, to provide some additional evidence to Uruguayan Spanish language about the relationships between oral language and cognitive development allows to carry out early interventions before formal schooling sets children for success in school and life.

Habilidades prelectoras en niños uruguayos de diferente nivel socioeconómico / Differential pre-literacy skills development in Uruguayan children from different socioeconomic backgrounds

Resumen El objetivo del presente estudio fue comparar las habilidades prelectoras en 50 preescolares uruguayos de 5 años de edad de diferente nivel socioeconómico (NSE) y analizar el impacto de estas habilidades en el aprendizaje de la lectura. Para ello, se evaluó a los niños mediante pruebas de vocabulario receptivo, conciencia fonológica, conocimiento sobre el nombre y el sonido de las letras, y denominación rápida de objetos a fin del nivel preescolar Tiempo 1 (T1). Un año más tarde, se evaluó a un subgrupo de la muestra inicial mediante una prueba de lectura de palabras Tiempo 2 (T2). Los resultados señalaron la existencia de correlaciones significativas entre los predictores (T1) y la lectura de palabras (T2) y entre todas las variables evaluadas y el nivel socioeconómico de los niños. La comparación del desempeño intergrupal señaló la existencia de diferencias significativas en todas las habilidades evaluadas a favor del nivel socioeconómico medio. Sin embargo, el desempeño en la lectura de palabras de ambos grupos fue bajo. Por otra parte, un análisis de regresión mostró que, para los niños de nivel socioeconómico bajo, el nivel de conciencia fonológica fue el que explicó la mayor parte de la varianza en la eficiencia lectora. El nivel de lectura de los niños de nivel socioeconómico medio fue mayormente explicado por el conocimiento del nombre de las letras. Los resultados ponen en evidencia la importancia de atender a las diferencias que se generan temprano en el desarrollo de habilidades lingüísticas fundamentales para aprender a leer.

Abstract Learning to read transforms lives. Reading contributes to knowledge acquisition, cultural engagement, and success in the school. The unequal distribution of literacy skills in a society is associated with economic and social inequalities as a result, children with a poor foundation in literacy before entering formal schooling are more likely to struggle academically and to drop out of school. For these reasons, there has been an intense scientific interest for decades in understanding how children learn to read. It is well established that in the early stages of reading development, phonological awareness, letter name-sounds knowledge, and the naming speed are three independent longitudinal predictors of children's later word-reading skills in alphabetic-writing systems. Phonological awareness constitutes the ability to recognize and manipulate the sounds of their own language, meanwhile letter knowledge promotes the discovery of systematic relationships between writing and oral language. As early readers develop some level of phonological awareness and some level of letter knowledge, they can recognize written words through phonological recoding processes, in which graphemes are recoded as phonemes and assembled to pronounce words. In addition, rapid naming expresses the speed at which phonological information is accessed from a graphic label. Phonological processing and letter knowledge are powerfully affected by the experience, stimulation, and support that children receive before beginning formal education. Most children acquire these abilities relatively effortlessly during early childhood. However, there is a significant number of children in Latin America who experience difficulties in their pre-literacy skills development. This study examined the cognitive profiles of a total of 50 Uruguayan preschoolers from different socioeconomic backgrounds from two public schools in Montevideo, Uruguay. Twenty-six children from low-income households were compared to peers from middle-income. At the end of the pre-schooling period (time 1) receptive vocabulary, phonological awareness, letter name-sounds knowledge, and object naming speed tests were administered to children. One year later (time 2), word -reading of a subgroup of children was measured. Significant correlations were observed between all predictors at time 1; between predictors at time 1 and word reading at time 2; and between all measured abilities and socioeconomic status. Comparative analysis between children of different socioeconomic status showed that children growing up in poverty contexts performed more poorly than their peers from middle-income families in all the tests. Nonetheless, both groups performed poorly in word reading. Descriptive statistics indicated that, out of a total of 26 words, low SES children correctly read a total of 7 words per minute, and medium SES children a total of 14 words. Finally, regression analyses indicated that phonological awareness contributed 30 % variance in predicting the total score achieved in a reading-word test in children of low-income families, meanwhile letter name knowledge contributed 74 % variance in predicting the total score achieved in a reading-word test in their peers from middle-income families. In general terms, results of pre-reading skills and reading performance seem to indicate that children of different socioeconomic status use different word recognition strategies according to their level of letter-knowledge of and phonological processing. Discussion considers international literature pointing out that children who enter elementary school with limited reading-related skills are unlikely to be able to keep pace with their peers. These findings warn about the importance to elaborate systematic and high-quality educational proposals to try to reduce the gap in reading development for children from low-income families. Developing literacy and language skills before formal schooling sets a child up for success in school and life. Results also suggest the importance of analyzing the variables that affect reading development in populations that are not the majority described.

Efficacy and Safety of COVID-19 Convalescent Plasma in Hospitalized Patients: A Randomized Clinical Trial.

Importance: There is clinical equipoise for COVID-19 convalescent plasma (CCP) use in patients hospitalized with COVID-19. Objective: To determine the safety and efficacy of CCP compared with placebo in hospitalized patients with COVID-19 receiving noninvasive supplemental oxygen. Design, Setting, and Participants: CONTAIN COVID-19, a randomized, double-blind, placebo-controlled trial of CCP in hospitalized adults with COVID-19, was conducted at 21 US hospitals from April 17, 2020, to March 15, 2021. The trial enrolled 941 participants who were hospitalized for 3 or less days or presented 7 or less days after symptom onset and required noninvasive oxygen supplementation. Interventions: A unit of approximately 250 mL of CCP or equivalent volume of placebo (normal saline). Main Outcomes and Measures: The primary outcome was participant scores on the 11-point World Health Organization (WHO) Ordinal Scale for Clinical Improvement on day 14 after randomization; the secondary outcome was WHO scores determined on day 28. Subgroups were analyzed with respect to age, baseline WHO score, concomitant medications, symptom duration, CCP SARS-CoV-2 titer, baseline SARS-CoV-2 serostatus, and enrollment quarter. Outcomes were analyzed using a bayesian proportional cumulative odds model. Efficacy of CCP was defined as a cumulative adjusted odds ratio (cOR) less than 1 and a clinically meaningful effect as cOR less than 0.8. Results: Of 941 participants randomized (473 to placebo and 468 to CCP), 556 were men (59.1%); median age was 63 years (IQR, 52-73); 373 (39.6%) were Hispanic and 132 (14.0%) were non-Hispanic Black. The cOR for the primary outcome adjusted for site, baseline risk, WHO score, age, sex, and symptom duration was 0.94 (95% credible interval [CrI], 0.75-1.18) with posterior probability (P[cOR<1] = 72%); the cOR for the secondary adjusted outcome was 0.92 (95% CrI, 0.74-1.16; P[cOR<1] = 76%). Exploratory subgroup analyses suggested heterogeneity of treatment effect: at day 28, cORs were 0.72 (95% CrI, 0.46-1.13; P[cOR<1] = 93%) for participants enrolled in April-June 2020 and 0.65 (95% CrI, 0.41 to 1.02; P[cOR<1] = 97%) for those not receiving remdesivir and not receiving corticosteroids at randomization. Median CCP SARS-CoV-2 neutralizing titer used in April to June 2020 was 1:175 (IQR, 76-379). Any adverse events (excluding transfusion reactions) were reported for 39 (8.2%) placebo recipients and 44 (9.4%) CCP recipients (P = .57). Transfusion reactions occurred in 2 (0.4) placebo recipients and 8 (1.7) CCP recipients (P = .06). Conclusions and Relevance: In this trial, CCP did not meet the prespecified primary and secondary outcomes for CCP efficacy. However, high-titer CCP may have benefited participants early in the pandemic when remdesivir and corticosteroids were not in use. Trial Registration: ClinicalTrials.gov Identifier: NCT04364737.

Functional convalescent plasma antibodies and pre-infusion titers shape the early severe COVID-19 immune response.

Transfer of convalescent plasma (CP) had been proposed early during the SARS-CoV-2 pandemic as an accessible therapy, yet trial results worldwide have been mixed, potentially due to the heterogeneous nature of CP. Here we perform deep profiling of SARS-CoV-2-specific antibody titer, Fc-receptor binding, and Fc-mediated functional assays in CP units, as well as in plasma from hospitalized COVID-19 patients before and after CP administration. The profiling results show that, although all recipients exhibit expanded SARS-CoV-2-specific humoral immune responses, CP units contain more functional antibodies than recipient plasma. Meanwhile, CP functional profiles influence the evolution of recipient humoral immunity in conjuncture with the recipient's pre-existing SARS-CoV2-specific antibody titers: CP-derived SARS-CoV-2 nucleocapsid-specific antibody functions are associated with muted humoral immune evolution in patients with high titer anti-spike IgG. Our data thus provide insights into the unexpected impact of CP-derived functional anti-spike and anti-nucleocapsid antibodies on the evolution of SARS-CoV-2-specific response following severe infection.

Single-Dilution COVID-19 Antibody Test with Qualitative and Quantitative Readouts.

The coronavirus disease 2019 (COVID-19) global pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to place an immense burden on societies and health care systems. A key component of COVID-19 control efforts is serological testing to determine the community prevalence of SARS-CoV-2 exposure and quantify individual immune responses to prior SARS-CoV-2 infection or vaccination. Here, we describe a laboratory-developed antibody test that uses readily available research-grade reagents to detect SARS-CoV-2 exposure in patient blood samples with high sensitivity and specificity. We further show that this sensitive test affords the estimation of viral spike-specific IgG titers from a single sample measurement, thereby providing a simple and scalable method to measure the strength of an individual's immune response. The accuracy, adaptability, and cost-effectiveness of this test make it an excellent option for clinical deployment in the ongoing COVID-19 pandemic.IMPORTANCE Serological surveillance has become an important public health tool during the COVID-19 pandemic. Detection of protective antibodies and seroconversion after SARS-CoV-2 infection or vaccination can help guide patient care plans and public health policies. Serology tests can detect antibodies against past infections; consequently, they can help overcome the shortcomings of molecular tests, which can detect only active infections. This is important, especially when considering that many COVID-19 patients are asymptomatic. In this study, we describe an enzyme-linked immunosorbent assay (ELISA)-based qualitative and quantitative serology test developed to measure IgG and IgA antibodies against the SARS-CoV-2 spike glycoprotein. The test can be deployed using commonly available laboratory reagents and equipment and displays high specificity and sensitivity. Furthermore, we demonstrate that IgG titers in patient samples can be estimated from a single measurement, enabling the assay's use in high-throughput clinical environments.

Functional Antibodies in COVID-19 Convalescent Plasma.

In the absence of an effective vaccine or monoclonal therapeutic, transfer of convalescent plasma (CCP) was proposed early in the SARS-CoV-2 pandemic as an easily accessible therapy. However, despite the global excitement around this historically valuable therapeutic approach, results from CCP trials have been mixed and highly debated. Unlike other therapeutic interventions, CCP represents a heterogeneous drug. Each CCP unit is unique and collected from an individual recovered COVID-19 patient, making the interpretation of therapeutic benefit more complicated. While the prevailing view in the field would suggest that it is administration of neutralizing antibodies via CCP that centrally provides therapeutic benefit to newly infected COVID-19 patients, many hospitalized COVID-19 patients already possess neutralizing antibodies. Importantly, the therapeutic benefit of antibodies can extend far beyond their simple ability to bind and block infection, especially related to their ability to interact with the innate immune system. In our work we deeply profiled the SARS-CoV-2-specific Fc-response in CCP donors, along with the recipients prior to and after CCP transfer, revealing striking SARS-CoV-2 specific Fc-heterogeneity across CCP units and their recipients. However, CCP units possessed more functional antibodies than acute COVID-19 patients, that shaped the evolution of COVID-19 patient humoral profiles via distinct immunomodulatory effects that varied by pre-existing SARS-CoV-2 Spike (S)-specific IgG titers in the patients. Our analysis identified surprising influence of both S and Nucleocapsid (N) specific antibody functions not only in direct antiviral activity but also in anti-inflammatory effects. These findings offer insights for more comprehensive interpretation of correlates of immunity in ongoing large scale CCP trials and for the design of next generation therapeutic design.

Treatment of severe COVID-19 with convalescent plasma in Bronx, NYC.

Convalescent plasma with severe acute respiratory disease coronavirus 2 (SARS-CoV-2) antibodies (CCP) may hold promise as a treatment for coronavirus disease 2019 (COVID-19). We compared the mortality and clinical outcome of patients with COVID-19 who received 200 mL of CCP with a spike protein IgG titer ≥ 1:2430 (median 1:47,385) within 72 hours of admission with propensity score-matched controls cared for at a medical center in the Bronx, between April 13 and May 4, 2020. Matching criteria for controls were age, sex, body mass index, race, ethnicity, comorbidities, week of admission, oxygen requirement, D-dimer, lymphocyte counts, corticosteroid use, and anticoagulation use. There was no difference in mortality or oxygenation between CCP recipients and controls at day 28. When stratified by age, compared with matched controls, CCP recipients less than 65 years had 4-fold lower risk of mortality and 4-fold lower risk of deterioration in oxygenation or mortality at day 28. For CCP recipients, pretransfusion spike protein IgG, IgM, and IgA titers were associated with mortality at day 28 in univariate analyses. No adverse effects of CCP were observed. Our results suggest CCP may be beneficial for hospitalized patients less than 65 years, but data from controlled trials are needed to validate this finding and establish the effect of aging on CCP efficacy.

Treatment of Severe COVID-19 with Convalescent Plasma in the Bronx, NYC.

Convalescent plasma with severe acute respiratory disease coronavirus 2 (SARS-CoV-2) antibodies (CCP) may hold promise as treatment for Coronavirus Disease 2019 (COVID-19). We compared the mortality and clinical outcome of patients with COVID-19 who received 200mL of CCP with a Spike protein IgG titer ≥1:2,430 (median 1:47,385) within 72 hours of admission to propensity score-matched controls cared for at a medical center in the Bronx, between April 13 to May 4, 2020. Matching criteria for controls were age, sex, body mass index, race, ethnicity, comorbidities, week of admission, oxygen requirement, D-dimer, lymphocyte counts, corticosteroids, and anticoagulation use. There was no difference in mortality or oxygenation between CCP recipients and controls at day 28. When stratified by age, compared to matched controls, CCP recipients <65 years had 4-fold lower mortality and 4-fold lower deterioration in oxygenation or mortality at day 28. For CCP recipients, pre-transfusion Spike protein IgG, IgM and IgA titers were associated with mortality at day 28 in univariate analyses. No adverse effects of CCP were observed. Our results suggest CCP may be beneficial for hospitalized patients <65 years, but data from controlled trials is needed to validate this finding and establish the effect of ageing on CCP efficacy.

Development, clinical translation, and utility of a COVID-19 antibody test with qualitative and quantitative readouts.

The COVID-19 global pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) continues to place an immense burden on societies and healthcare systems. A key component of COVID-19 control efforts is serologic testing to determine the community prevalence of SARS-CoV-2 exposure and quantify individual immune responses to prior infection or vaccination. Here, we describe a laboratory-developed antibody test that uses readily available research-grade reagents to detect SARS-CoV-2 exposure in patient blood samples with high sensitivity and specificity. We further show that this test affords the estimation of viral spike-specific IgG titers from a single sample measurement, thereby providing a simple and scalable method to measure the strength of an individual's immune response. The accuracy, adaptability, and cost-effectiveness of this test makes it an excellent option for clinical deployment in the ongoing COVID-19 pandemic.

The Fungal Cell Wall: Candida, Cryptococcus, and Aspergillus Species.

The fungal cell wall is located outside the plasma membrane and is the cell compartment that mediates all the relationships of the cell with the environment. It protects the contents of the cell, gives rigidity and defines the cellular structure. The cell wall is a skeleton with high plasticity that protects the cell from different stresses, among which osmotic changes stand out. The cell wall allows interaction with the external environment since some of its proteins are adhesins and receptors. Since, some components have a high immunogenic capacity, certain wall components can drive the host's immune response to promote fungus growth and dissemination. The cell wall is a characteristic structure of fungi and is composed mainly of glucans, chitin and glycoproteins. As the components of the fungal cell wall are not present in humans, this structure is an excellent target for antifungal therapy. In this article, we review recent data on the composition and synthesis, influence of the components of the cell wall in fungi-host interaction and the role as a target for the next generation of antifungal drugs in yeasts (Candida and Cryptococcus) and filamentous fungi (Aspergillus).

Diffusing Capacity of the Lung for Carbon Monoxide in Mexican/Latino Children. Quality Control and Reference Values.

RATIONALE: Single-breath diffusing capacity of the lung for carbon monoxide (DlCOsb) values are used to evaluate gas exchange; however, the quality of maneuvers performed by children has not been evaluated, and reference values for young people living at moderate altitudes are not well established. OBJECTIVES: Our objectives were 1) to determine whether DlCOsb maneuvers performed by a pediatric population would meet 2017 European Respiratory Society/American Thoracic Society (ERS/ATS) quality control standards; and 2) to report normal DlCOsb values for Mexican/Latino children and adolescents living at moderate altitudes. METHODS: This study involved healthy young people 4-20 years of age from the metropolitan area of Mexico City (2,240 m above sea level) who were recruited in schools from July 2014 to August 2017. DlCOsb testing was performed according to the 2005 ATS/ERS standards, and the quality control of each maneuver was analyzed according to the 2017 ERS/ATS standards. We constructed models for DlCOsb with linear and quadratic terms for weight, height, and age as independent variables using shrinkage statistics, variance inflation factors, the Akaike information criterion, and R2 to compare the results of different models. RESULTS: Results were obtained for 420 individuals (53% boys) with a mean age of 11.7 ± 4.5 standard deviation (SD) years; 47% of maneuvers from children age 4-6 years were grade A (13% grade B), and 90% of those in children older than 13 years were grade A or B. Forty-six percent of the subjects had a DlCOsb repeatability of <1 ml/min/mm Hg. The mean DlCOsb was higher for boys than for girls (32.4 ± 13.6 [SD] vs. 24.1 ± 7.5 ml/min/mm Hg, respectively). The reference equation for boys was DlCOsb = exp(1.63469 + [0.03251 × age] + [0.00846 × height] + [0.00304 × weight]), R2 = 0.87; for girls, the best equation was DlCOsb = exp(1.56516 + [0.0193 × age] + [0.00893 × height] + [0.00273 × weight]), R2 = 0.75. The single-breath transfer coefficient of the lung for carbon monoxide remained constant with age and height, with a lower limit of normal of 6.5 ml/min/mm Hg/L in boys and 5.4 ml/min/mm Hg/L in girls. Measured DlCOsb was higher than predicted by other authors (P < 0.001 by paired t test). CONCLUSIONS: Individuals 4-20 years of age can complete high-quality DlCOsb tests. Children and adolescents living at 2,240 m have higher DlCOsb values than those living at sea level. Reference equations for DlCOsb obtained at sea level are poor predictors of the values measured at moderate altitude.

Serum-Mediated Cleavage of Bacillus anthracis Protective Antigen Is a Two-Step Process That Involves a Serum Carboxypeptidase.

Much of our understanding of the activity of anthrax toxin is based on in vitro systems, which delineate the interaction between Bacillus anthracis toxins and the cell surface. However, these systems fail to account for the intimate association of B. anthracis with the circulatory system, including the contribution of serum proteins to the host response and processing of anthrax toxins. Using a variety of immunological techniques to inhibit serum processing of B. anthracis protective antigen (PA) along with mass spectrometry analysis, we demonstrate that serum digests PA via 2 distinct reactions. In the first reaction, serum cleaves PA83 into 2 fragments to produce PA63 and PA20 fragments, similarly to that observed following furin digestion. This is followed by carboxypeptidase-mediated removal of the carboxy-terminal arginine and lysines from PA20IMPORTANCE Our findings identify a serum-mediated modification of PA20 that has not been previously described. These observations further imply that the processing of PA is more complex than currently thought. Additional study is needed to define the contribution of serum processing of PA to the host response and individual susceptibility to anthrax.

Naïve B cells reduce fungal dissemination in Cryptococcus neoformans infected Rag1-/- mice.

IgM and B-1 cell deficient mice exhibit early C. neoformans dissemination from lungs to brain, but a definitive role for B cells in conferring resistance to C. neoformans dissemination has not been established. To address this question, we developed an intranasal (i.n.) C. neoformans infection model in B and T cell deficient Rag1-/- mice and found they also exhibit earlier fungal dissemination and higher brain CFU than wild-type C57Bl/6 (wild-type) mice. To probe the effect of B cells on fungal dissemination, Rag1-/- mice were given splenic (intravenously) or peritoneal (intraperitoneally) B cells from wild-type mice and infected i.n. with C. neoformans 7 d later. Mice that received B cells had lung histopathology resembling wild type mice 14 d post-infection, and B-1, not B-2 or T cells in their lungs, and serum and lung IgM and IgG 21 d post-infection. Lung CFU were comparable in wild-type, Rag1-/-, and Rag1-/- mice that received B cells 21 d post-infection, but brain CFU were significantly lower in mice that received B cells than Rag1-/- mice that did not. To determine if natural antibody can promote immunity in our model, we measured alveolar macrophage phagocytosis of C. neoformans in Rag1-/- mice treated with naive wild-type IgM-sufficient or sIgM-/- IgM-deficient sera before infection. Compared to IgM-deficient sera, IgM-sufficient sera significantly increased phagocytosis. Our data establish B cells are able to reduce early C. neoformans dissemination in mice and suggest natural IgM may be a key mediator of early antifungal immunity in the lungs.

A Small Protein Associated with Fungal Energy Metabolism Affects the Virulence of Cryptococcus neoformans in Mammals.

The pathogenic yeast Cryptococcus neoformans causes cryptococcosis, a life-threatening fungal disease. C. neoformans has multiple virulence mechanisms that are non-host specific, induce damage and interfere with immune clearance. Microarray analysis of C. neoformans strains serially passaged in mice associated a small gene (CNAG_02591) with virulence. This gene, hereafter identified as HVA1 (hypervirulence-associated protein 1), encodes a protein that has homologs of unknown function in plant and animal fungi, consistent with a conserved mechanism. Expression of HVA1 was negatively correlated with virulence and was reduced in vitro and in vivo in both mouse- and Galleria-passaged strains of C. neoformans. Phenotypic analysis in hva1Δ and hva1Δ+HVA1 strains revealed no significant differences in established virulence factors. Mice infected intravenously with the hva1Δ strain had higher fungal burden in the spleen and brain, but lower fungal burden in the lungs, and died faster than mice infected with H99W or the hva1Δ+HVA1 strain. Metabolomics analysis demonstrated a general increase in all amino acids measured in the disrupted strain and a block in the TCA cycle at isocitrate dehydrogenase, possibly due to alterations in the nicotinamide cofactor pool. Macrophage fungal burden experiments recapitulated the mouse hypervirulent phenotype of the hva1Δ strain only in the presence of exogenous NADPH. The crystal structure of the Hva1 protein was solved, and a comparison of structurally similar proteins correlated with the metabolomics data and potential interactions with NADPH. We report a new gene that modulates virulence through a mechanism associated with changes in fungal metabolism.

The Membrane Phospholipid Binding Protein Annexin A2 Promotes Phagocytosis and Nonlytic Exocytosis of Cryptococcus neoformans and Impacts Survival in Fungal Infection.

Cryptococcus neoformans is a fungal pathogen with a unique intracellular pathogenic strategy that includes nonlytic exocytosis, a phenomenon whereby fungal cells are expunged from macrophages without lysing the host cell. The exact mechanism and specific proteins involved in this process have yet to be completely defined. Using murine macrophages deficient in the membrane phospholipid binding protein, annexin A2 (ANXA2), we observed a significant decrease in both phagocytosis of yeast cells and the frequency of nonlytic exocytosis. Cryptococcal cells isolated from Anxa2-deficient (Anxa2(-/-)) bone marrow-derived macrophages and lung parenchyma displayed significantly larger capsules than those isolated from wild-type macrophages and tissues. Concomitantly, we observed significant differences in the amount of reactive oxygen species produced between Anxa2(-/-) and Anxa2(+/+) macrophages. Despite comparable fungal burden, Anxa2(-/-) mice died more rapidly than wild-type mice when infected with C. neoformans, and Anxa2(-/-) mice exhibited enhanced inflammatory responses, suggesting that the reduced survival reflected greater immune-mediated damage. Together, these findings suggest a role for ANXA2 in the control of cryptococcal infection, macrophage function, and fungal morphology.

Platelet-activating factor contributes to Bacillus anthracis lethal toxin-associated damage.

The lethal toxin (LeTx) of Bacillus anthracis plays a central role in the pathogenesis of anthrax-associated shock. Platelet-activating factor (PAF) is a potent lipid mediator that has been implicated in endotoxin-associated shock. In this study, we examined the contribution of PAF to the manifestations of lethal toxin challenge in WT mice. LeTx challenge resulted in transient increase in serum PAF levels and a concurrent decrease in PAF acetylhydrolase activity. Inhibition of PAF activity using PAF antagonists or toxin challenge of PAF receptor negative mice reversed or ameliorated many of the pathologic features of LeTx-induced damage, including changes in vascular permeability, hepatic necrosis, and cellular apoptosis. In contrast, PAF inhibition had minimal effects on cytokine levels. Findings from these studies support the continued study of PAF antagonists as potential adjunctive agents in the treatment of anthrax-associated shock.

Microbicidal power of alpha radiation in sterilizing germinating Bacillus anthracis spores.

Radioimmunotherapy (RIT) takes advantage of the specificity and affinity of the antigen-antibody interaction to deliver microbicidal radioactive nuclides to a site of infection. In this study, we investigated the microbicidal properties of an alpha particle-emitting 213Bi-labeled monoclonal antibody (MAb), EA2-1 (213Bi-EA2-1), that binds to the immunodominant antigen on Bacillus anthracis spores. Our results showed that dormant spores were resistant to 213Bi-EA2-1. Significant spore killing was observed following treatment with EA2-1 labeled with 300 µCi 213Bi; however, this effect was not dependent on the MAb. In contrast, when spores were germinating, 213Bi-EA2-1 mediated MAb-specific killing in a dose-dependent manner. Dormant spores are very resistant to RIT, and RIT should focus on targeting vegetative cells and germinating spores.