Current advances in experimental and computational approaches to enhance CAR T cell manufacturing protocols and improve clinical efficacy.

Since the FDA's approval of chimeric antigen receptor (CAR) T cells in 2017, significant improvements have been made in the design of chimeric antigen receptor constructs and in the manufacturing of CAR T cell therapies resulting in increased in vivo CAR T cell persistence and improved clinical outcome in certain hematological malignancies. Despite the remarkable clinical response seen in some patients, challenges remain in achieving durable long-term tumor-free survival, reducing therapy associated malignancies and toxicities, and expanding on the types of cancers that can be treated with this therapeutic modality. Careful analysis of the biological factors demarcating efficacious from suboptimal CAR T cell responses will be of paramount importance to address these shortcomings. With the ever-expanding toolbox of experimental approaches, single-cell technologies, and computational resources, there is renowned interest in discovering new ways to streamline the development and validation of new CAR T cell products. Better and more accurate prognostic and predictive models can be developed to help guide and inform clinical decision making by incorporating these approaches into translational and clinical workflows. In this review, we provide a brief overview of recent advancements in CAR T cell manufacturing and describe the strategies used to selectively expand specific phenotypic subsets. Additionally, we review experimental approaches to assess CAR T cell functionality and summarize current in silico methods which have the potential to improve CAR T cell manufacturing and predict clinical outcomes.

Síndrome de burnout en pandemia y satisfacción laboral en médicos serumistas de Ayacucho / Pandemic burnout and job satisfaction among doctors from Ayacucho engaged in the SERUMS program

RESUMEN Objetivo: Evaluar la asociación entre el síndrome de burnout (SB) y la satisfacción laboral (SL) en los médicos serumistas de Ayacucho en el contexto de la pandemia por la COVID-19. Materiales y métodos: Estudio observacional, transversal, que usó un modelo crudo y ajustado para evaluar la asociación entre el SB, valorado por medio del cuestionario Maslach Burnout Inventory (MBI), y la SL, por medio del cuestionario de satisfacción S20/23. Las asociaciones se presentaron como razón de prevalencia (RP) con sus respectivos intervalos de confianza al 95 % (IC al 95 %). Resultados: De los 70 médicos serumistas, se encontró que el 77,14 % tenía entre 18-33 años, el 52,86 % eran hombres, el 51,43 % tenía menos de un año de experiencia laboral y el 88,57 % laboró en un centro de salud del Ministerio de Salud (Minsa). La prevalencia del SB fue de 45,71 %. La prevalencia del SB con insatisfacción fue del 30 %; con satisfacción, 32,86 %, y los indiferentes, 37,14 %. En el análisis crudo encontramos significancia estadística entre el SB y la SL (RP = 13,5; IC del 95 %:3,08-59,24), y en el análisis ajustado por sexo, estado civil y tiempo de experiencia laboral, la asociación entre el SB y la SL permaneció estadísticamente significativa (RP = 14,15; IC del 95 %: 3,02-66,32). Conclusiones: El SB se asocia a la SL de manera negativa, encontrando uno de cada dos médicos con SB. Además, se sugiere que existe mayor probabilidad de insatisfacción laboral en el personal con SB. Ello da a entender que podría usarse como indicador en diferentes áreas de la gestión y ser un punto a evaluar en la toma de decisiones en las directivas de salud. La solución de estos problemas contribuiría a mejorar las condiciones laborales actuales y, por ende, el sistema de salud.

ABSTRACT Objective: To determine the association between psychosocial work factors and symptoms of anxiety or depression among healthcare personnel in the Intensive Care Unit (ICU) at Hospital Regional Eleazar Guzmán Barrón in Nuevo Chimbote. Materials and methods: An observational, cross-sectional study was conducted, using both crude and adjusted models, to determine the association between burnout syndrome (BS), evaluated through the Maslach Burnout Inventory (MBI), and job satisfaction (JS), assessed via the Job Satisfaction Questionnaire S20/23. The associations were expressed as prevalence ratios (PR) with their respective 95 % confidence intervals (95 % CI). Results: Out of the 70 doctors engaged in the SERUMS program, 77.14 % were aged between 18 and 33 years, 52.86 % were males, 51.43 % had less than one year work experience and 88.57 % worked at a health center of the Ministry of Health. The prevalence of BS was 45.71 %. The prevalence of JS was as follows: dissatisfaction 30 %, satisfaction 32.86 % and indifference 37.14 %. In the crude analysis, a statistically significant association between BS and JS was found (PR = 13.5; 95 % CI: 3.08 59.24) and in the analysis adjusted for sex, marital status and length of work experience, the association between BS and JS remained statistically significant (PR = 14.15; 95 % CI: 3.02 66.32). Conclusions: BS is negatively associated with JS, with one in two doctors experiencing BS. Moreover, there appears to be a higher likelihood of job dissatisfaction among personnel with BS. This suggests that BS could serve as an indicator in different management areas and be considered in health management decision-making. Solving these issues could contribute to improving current working conditions and, therefore, the healthcare system.

Utilizing immunogenomic approaches to prioritize targetable neoantigens for personalized cancer immunotherapy.

Advancements in sequencing technologies and bioinformatics algorithms have expanded our ability to identify tumor-specific somatic mutation-derived antigens (neoantigens). While recent studies have shown neoantigens to be compelling targets for cancer immunotherapy due to their foreign nature and high immunogenicity, the need for increasingly accurate and cost-effective approaches to rapidly identify neoantigens remains a challenging task, but essential for successful cancer immunotherapy. Currently, gene expression analysis and algorithms for variant calling can be used to generate lists of mutational profiles across patients, but more care is needed to curate these lists and prioritize the candidate neoantigens most capable of inducing an immune response. A growing amount of evidence suggests that only a handful of somatic mutations predicted by mutational profiling approaches act as immunogenic neoantigens. Hence, unbiased screening of all candidate neoantigens predicted by Whole Genome Sequencing/Whole Exome Sequencing may be necessary to more comprehensively access the full spectrum of immunogenic neoepitopes. Once putative cancer neoantigens are identified, one of the largest bottlenecks in translating these neoantigens into actionable targets for cell-based therapies is identifying the cognate T cell receptors (TCRs) capable of recognizing these neoantigens. While many TCR-directed screening and validation assays have utilized bulk samples in the past, there has been a recent surge in the number of single-cell assays that provide a more granular understanding of the factors governing TCR-pMHC interactions. The goal of this review is to provide an overview of existing strategies to identify candidate neoantigens using genomics-based approaches and methods for assessing neoantigen immunogenicity. Additionally, applications, prospects, and limitations of some of the current single-cell technologies will be discussed. Finally, we will briefly summarize some of the recent models that have been used to predict TCR antigen specificity and analyze the TCR receptor repertoire.

Delivering an Immunocastration Vaccine via a Novel Subcutaneous Implant.

Immunocastration relies on the vaccine-mediated stimulation of an immune response to gonadotropin-releasing hormone (GnRH) in order to interrupt spermatogenesis. This approach offers a less painful alternative to traditional castration approaches but the current, commercially available options require multiple doses of vaccine to maintain sterility. Thus, a series of pilot studies were conducted to determine the feasibility of a single-dose immunocastration vaccine implant. These five studies utilized a total of 44 Holstein bulls to determine the optimal vaccine composition and validate the ability of a stainless-steel subcutaneous implant to deliver a vaccine. Outcome measures included the duration of implant retention, scrotal dimensions and temperature, implant site temperature, anti-GnRH antibodies, and serum testosterone concentration. Over the course of several studies, anti-GnRH antibodies were successfully stimulated by vaccine implants. No significant treatment effects on scrotal dimensions or testosterone were detected over time, but changes in spermatogenesis were detected across treatment groups. Results indicate that a single-dose implantable immunocastration vaccine elicits a humoral immune response and could impact spermatogenesis in bulls. These findings provide opportunities for the refinement of this technology to improve implant retention over longer periods of time. Taken together, this approach will offer producers and veterinarians an alternative to physical castration methods, to improve animal welfare during routine livestock management procedures.

Presentación neurológica severa en COVID-19 / Severe neurological presentation in COVID-19

Se presenta el caso de un varón de 53 años de edad con un tiempo de enfermedad de tres días, con malestar general, fiebre, dolor faríngeo, y tos seca. Al ingresar al hospital, se le halló febril y presentó convulsiones tónicas-clónicas y desorientación. La tomografía pulmonar mostró lesiones compatibles con COVID-19; TAC cerebral normal; y, rT-PCR para COVID-19: positivo. La punción lumbar mostró solo hiperglucorraquia, El paciente falleció al tercer día. Se concluye que se trató de un paciente con COVID-19 y compromiso neurológico tipo encefalitis. (AU)

This is the case of a 53-year-old male with a three-day disease time, with malaise, fever, pharyngeal pain and dry cough. Upon entering the hospital, he was febrile and had tonic-clonic seizures and disorientation. The pulmonary tomography showed COVID-19-compatible lesions; normal brain CT; and, rT-PCR for COVID-19: positive. The lumbar puncture showed only hyperglucorraquia, The patient died on the third day. It is concluded that it was a patient with COVID-19 and neurological involvement type encephalitis. (AU)

Characterizing the soil microbiome and quantifying antibiotic resistance gene dynamics in agricultural soil following swine CAFO manure application.

As agriculture industrializes, concentrated animal feeding operations (CAFOs) are becoming more common. Feces from CAFOs is often used as fertilizer on fields. However, little is known about the effects manure has on the soil microbiome, which is an important aspect of soil health and fertility. In addition, due to the subtherapeutic levels of antibiotics necessary to keep the animals healthy, CAFO manure has elevated levels of antibiotic resistant bacteria. Using 16s rRNA high-throughput sequencing and qPCR, this study sought to determine the impact of swine CAFO manure application on both the soil microbiome and abundance of select antibiotic resistance genes (ARGs) and mobile element genes (erm(B), erm(C), sul1, str(B), intI1, IncW repA) in agricultural soil over the fall and spring seasons. We found the manure community to be distinct from the soil community, with a majority of bacteria belonging to Bacteroidetes and Firmicutes. The soil samples had more diverse communities dominated by Acidobacteria, Actinobacteria, Proteobacteria, Verrucomicrobia, and unclassified bacteria. We observed significant differences in the soil microbiome between all time points, except between the spring samples. However, by tracking manure associated taxa, we found the addition of the manure microbiome to be a minor driver of the shift. Of the measured genes, manure application only significantly increased the abundance of erm(B) and erm(C) which remained elevated in the spring. These results suggest bacteria in the manure do not survive well in soil and that ARG dynamics in soil following manure application vary by resistance gene.

Water for African elephants (Loxodonta africana): faecal microbial loads affect use of artificial waterholes.

In semi-arid protected areas, artificial waterholes ensure that water is locally available to animals for extended periods. However, artificial waterholes may limit animal movement, which contributes towards habitat deterioration. Challenges of artificial water provisioning worsen in the presence of ecosystem engineers like African elephants Loxodonta africana, capable of transforming environments. Camera traps were used to monitor elephant visitation at 21 artificial waterholes in the Kruger National Park, South Africa. We also assessed if water quality parameters influenced elephant preference for certain waterholes. There were no significant correlations between elephant abundance and water physicochemical properties. However, there was a strong negative correlation between elephant abundance and levels of Escherichia coli in water. Our findings suggest that elephants avoid drinking water with high levels of faecal microbial loads. Whereas most studies addressing animal management in protected areas consider waterholes as homogeneous units, we posit that water quality could also determine local landscape use and movement patterns of key species like elephants, a finding with relevant implications in reserve management practices.