Tumor intrinsic activity of Chromobox 2 (CBX2) remodels the tumor microenvironment in high grade serous carcinoma.

Chromobox 2 (CBX2), an epigenetic reader and component of Polycomb Repressor Complex 1 (PRC1), is highly expressed in >75% of high-grade serous carcinoma (HGSC). Increased CBX2 expression is associated with poorer survival, while CBX2 knockdown leads to improved chemotherapy sensitivity. In an HGSC immune competent murine model, knockdown of CBX2 decreased tumor progression. We sought to explore the impact of modulation of CBX2 on the tumor immune microenvironment (TIME), understanding that the TIME plays a critical role in disease progression and development of therapy resistance. Exploration of existing datasets demonstrated that elevated CBX2 expression significantly correlated with the specific immune cell types in the TIME. RNA-seq and pathway analysis of differentially expressed genes demonstrated immune signature enrichment. Confocal microscopy and co-culture experiments found modulation of CBX2 leads to increased recruitment and infiltration of macrophages. Flow cytometry of macrophages cultured with CBX2 overexpressing cells showed increased M2-like macrophages and decreased phagocytosis activity. Cbx2 knockdown in the Trp53, Brca2 null ID8 syngeneic murine model (ID8 Trp53-/- Brca2-/-) led to decreased tumor progression compared to control. NanoString Immuno-Oncology Panel analysis suggested knock down in Cbx2 shifts immune cell composition, with an increase in macrophages. Multispectral immunohistochemistry further confirmed an increase in macrophage infiltration. Increased CBX2 expression leads to recruitment and polarization of pro-tumor macrophages and targeting CBX2 may serve to modulate the TIME to enhance the efficacy of immune therapies.

Characterization of Novel Trypanosoma cruzi-Specific Antigen with Potential Use in the Diagnosis of Chagas Disease.

Chagas disease is caused by the parasite Trypanosoma cruzi. In humans, it evolves into a chronic disease, eventually resulting in cardiac, digestive, and/or neurological disorders. In the present study, we characterized a novel T. cruzi antigen named Tc323 (TcCLB.504087.20), recognized by a single-chain monoclonal antibody (scFv 6B6) isolated from the B cells of patients with cardiomyopathy related to chronic Chagas disease. Tc323, a ~323 kDa protein, is an uncharacterized protein showing putative quinoprotein alcohol dehydrogenase-like domains. A computational molecular docking study revealed that the scFv 6B6 binds to an internal domain of Tc323. Immunofluorescence microscopy and Western Blot showed that Tc323 is expressed in the main developmental forms of T. cruzi, localized intracellularly and exhibiting a membrane-associated pattern. According to phylogenetic analysis, Tc323 is highly conserved throughout evolution in all the lineages of T. cruzi so far identified, but it is absent in Leishmania spp. and Trypanosoma brucei. Most interestingly, only plasma samples from patients infected with T. cruzi and those with mixed infection with Leishmania spp. reacted against Tc323. Collectively, our findings demonstrate that Tc323 is a promising candidate for the differential serodiagnosis of chronic Chagas disease in areas where T. cruzi and Leishmania spp. infections coexist.

The programmed death ligand 1 interactome demonstrates bidirectional signaling coordinating immune suppression and cancer progression in head and neck squamous cell carcinoma.

BACKGROUND: The programmed cell death protein 1 (PD-1) and programmed death ligand 1 (PD-L1) are validated cancer targets; however, emerging mechanisms and impact of PD-L1 intracellular signaling on cancer behavior are poorly understood. METHODS: We investigated the cancer cell intrinsic role of PD-L1 in multiple patient-derived models in vitro and in vivo. PD-L1 overexpression, knockdown, and PD-L1 intracellular domain (PD-L1-ICD) deletion (Δ260-290PD-L1) models were assessed for key cancer properties: clonogenicity, motility, invasion, and immune evasion. To determine how PD-L1 transduces signals intracellularly, we used the BioID2 platform to identify the PD-L1 intracellular interactome. Both human papillomavirus-positive and negative patient-derived xenografts were implanted in NOD-scid-gamma and humanized mouse models to investigate the effects of recombinant PD-1, anti-PD-L1, and anti-signal transducer and activator of transcription 3 (STAT3) in vivo. RESULTS: PD-L1 intracellular signaling increased clonogenicity, motility, and invasiveness in multiple head and neck squamous cell carcinoma (HNSCC) models, and PD-1 binding enhanced these effects. Protein proximity labeling revealed the PD-L1 interactome, distinct for unbound and bound PD-1, which initiated cancer cell-intrinsic signaling. PD-L1 binding partners interleukin enhancer binding factors 2 and 3 (ILF2-ILF3) transduced their effect through STAT3. Δ260-290PD-L1 disrupted signaling and reversed pro-growth properties. In humanized HNSCC in vivo models bearing T-cells, PD-1 binding triggered PD-L1 signaling, and dual PD-L1 and STAT3 inhibition were required to achieve tumor control. CONCLUSIONS: Upon PD-1 binding, the PD-L1 extracellular and intracellular domains exert a synchronized effect to promote immune evasion by inhibiting T-cell function while simultaneously enhancing cancer cell-invasive properties.

Endogenous Retrovirus RNA Expression Differences between Race, Stage and HPV Status Offer Improved Prognostication among Women with Cervical Cancer.

Endogenous human retroviruses (ERVs) are remnants of exogenous retroviruses that have integrated into the human genome. Using publicly available RNA-seq data from 63 cervical cancer patients, we investigated the expression of ERVs in cervical cancers. Four aspects of cervical cancer were investigated: patient ancestral background, tumor HPV type, tumor stage and patient survival. Between the racial subgroups, 74 ERVs were significantly differentially expressed, with Black Americans having 30 upregulated and 44 downregulated (including MER21C, HERV9-int, and HERVH-int) ERVs when compared to White Americans. We found that 3313 ERVs were differentially expressed between HPV subgroups, including MER41A, HERVH-int and HERVK9. There were 28 downregulated (including MLT1D and HERVH-int) and 61 upregulated (including MER41A) ERVs in locally advanced-stage compared to early-stage samples. Tissue microarrays of cervical cancer patients were used to investigate the protein expression of ERVs with protein coding potential (i.e., HERVK and ERV3). Significant differences in protein expression of ERV3 (p = 0.000905) were observed between early-stage and locally advanced-stage tumors. No significant differential expression at the protein level was found for HERVK7 (p = 0.243). We also investigated a prognostic model, supplementing a baseline prediction model using FIGO stage, age and HPV positivity with ERVs data. The expression levels of all ERVs in the HERVd were input into a Lasso-Cox proportional hazards model, developing a predictive 67-ERV panel. When ERVs expression levels were supplemented with the clinical data, a significant increase in prognostic power (p = 9.433 × 10-15) relative to that obtained with the clinical parameters alone (p = 0.06027) was observed. In summary, ERV RNA expression in cervical cancer tumors is significantly different among racial cohorts, HPV subgroups and disease stages. The combination of the expression of certain ERVs in cervical cancers with clinical factors significantly improved prognostication compared to clinical factors alone; therefore, ERVs may serve as future prognostic biomarkers and therapeutic targets. Novelty and Impact: When endogenous retroviral (ERV) expression signatures were combined with currently employed clinical prognosticators of relapse of cervical cancer, the combination outperformed prediction models based on clinical prognosticators alone. ERV expression signatures in tumor biopsies may therefore be useful to help identify patients at greater risk of recurrence. The novel ERV expression signatures or adjacent genes possibly impacted by ERV expression described here may also be targets for the development of future therapeutic interventions.

Expression of Inhibitory Receptors TIGIT, TIM-3, and LAG-3 on CD4+ T Cells from Patients with Different Clinical Forms of Chronic Chagas Disease.

T cells are central to the adaptive immune response against Trypanosoma cruzi infection. In chronic Chagas disease (CCD), circulating parasite-specific memory T cells show reduced functionality and increased expression of inhibitory receptors as a result of persistent antigenic stimulation. This phenotype has been linked to progression of cardiac pathology, whereas the presence of polyfunctional T cells shows association with therapeutic success. In this study, we demonstrate that T. cruzi-specific human CD4+ T cells can be identified by their expression of OX40 and CD25 upon in vitro stimulation. We characterized the expression of the inhibitory receptors T cell immunoreceptor with Ig and ITIM domains (TIGIT), T cell Ig and mucin-domain containing-3 (TIM-3), and lymphocyte activation gene 3 (LAG-3) in CD4+ T cells from CCD patients with and without cardiac alterations. Our results show that, independently of their clinical stage, CCD patients present an increased frequency of CD4+ T cells expressing TIGIT in comparison with non-T. cruzi-infected donors. Exposure to parasite Ags increases the expression of TIM-3 in CD4+ T cells from CCD patients, especially in those with cardiac compromise. Upregulation of LAG-3 was also detected in CCD individuals without cardiac manifestations, predominantly within the subpopulation of cells that did not become activated upon stimulation. Further differences were found between groups in the coexpression of these receptors. Blockade of each individual receptor did not affect activation or the production of IFN-γ and IL-10 by CD4+ T cells in response to parasite Ags. Our results suggest a role for TIGIT, TIM-3, and LAG-3 in the modulation of inflammatory phenomena thought to ultimately lead to tissue damage and cardiac pathology.

Activation-induced marker assays for identification of Trypanosoma cruzi-specific CD4 or CD8 T cells in chronic Chagas disease patients.

Antigen-specific T cells are central to the adaptive immune response against T. cruzi infection and underpin the efficacy of on-going vaccine strategies. In this context, the present study focuses on T-cell assays that define the parasite-specificity on the basis of upregulation of TCR stimulation-induced surface markers. For this purpose, we tested different dual marker combinations (OX40, CD25, CD40L, CD137, CD69, PD-L1, CD11a, CD49d, HLA-DR, CD38) to reliably identify activated CD4+ and CD8+ T-cell populations from PBMCs of chronic Chagas disease (CCD) patients after 12 or 24 h of stimulation with T. cruzi lysate. Results demonstrated that activation-induced markers (AIM) assays combining the expression of OX40, CD25, CD40L, CD137, CD69 and/or PD-L1 surface markers are efficient at detecting T. cruzi-specific CD4+ T cells in CCD patients, in comparison to non-infected donors, after both stimulation times. For CD8+ T cells, only PD-L1/OX40 after 24 h of antigen exposure resulted to be useful to track a parasite-specific response. We also demonstrated that the agnostic activation is mediated by different T. cruzi strains, such as Dm28c, CL Brener or Sylvio. Additionally, we successfully used this approach to identify the phenotype of activated T lymphocytes based on the expression of CD45RA and CCR7. Overall, our results show that different combinations of AIM markers represent an effective and simple tool for the detection of T. cruzi-specific CD4+ and CD8+ T cells.

Cardiovascular risk in hashimoto thyroiditis patients according to their thyroid state: a cross-sectional study / Riesgo cardiovascular en pacientes con tiroiditis hashimoto según su estada tiroidea: un estudio transversal

Objetivo: investigar la asociación entre los factores de riesgo cardiovascular y el nivel de riesgo cardiovascular con el estado tiroideo de pacientes con Tiroiditis de Hashimoto. Métodos y técnicas: treinta y ocho adultos con tiroiditis de Hashimoto participaron en este estudio descriptivo transversal. Los factores de riesgo cardiovascular considerados incluyeron edad, sexo, presión arterial, índice de masa corporal, glucemia, perfil lipídico, comorbilidades cardiovasculares, Proteína C Reactiva y Eritrosedimentación. Se utilizó la calculadora de riesgo cardiovascular de Framingham. La muestra se clasificó según el estado tiroideo en eutiroidismo (n = 15), hipotiroidismo clínico (n = 9) e hipotiroidismo subclínico (n = 13) e incluyó la presencia de anticuerpos antitiroideos. Se utilizó la prueba exacta de Fisher para determinar la asociación entre las variables estudiadas. Resultados: el 100 % de la muestra fue de sexo femenino; con una edad media entre 39-59 años. La categoría con bajo riesgo fue la mayor muestra (n = 30), equivalente al 78,9 %; riesgo moderado, no se obtuvo ningún paciente; alto riesgo (n = 8) constituyó el 21,1 %. Se encontró significancia estadística entre la edad y el nivel de riesgo cardiovascular en pacientes con hipotiroidismo clínico (p < 1), score in patients with clinical hypothyroidism was found (p < 1), IC 95 %. El nivel de glucosa en el hipotiroidismo subclínico y el hipotiroidismo clínico tuvo significancia estadística. Del mismo modo, se demostró que la presencia de antitiroglobulina (antiTg) está estrechamente relacionada con el nivel de riesgo cardiovascular en pacientes con hipotiroidismo subclínico. Conclusión: la edad, la glucemia, los anti-Tg, el antecedente de diabetes mellitus, la dislipemia y haber padecido algún accidente cerebrovascular se ha relacionado con un aumento del riesgo de desarrollar un evento cardiovascular hasta en 10 años en función de su perfil tiroideo. No se encontró evidencia de una relación directa entre la puntuación de riesgo cardiovascular y el estado de la tiroides en los participantes de este estudio.

Aims: To investigate the association between cardiovascular risk factors and cardiovascular risk score with the thyroid status of patients with Hashimoto's Thyroiditis. Methods: Thirty-eight consenting adults with Hashimoto's Thyroiditis participated in this cross-sectional study. The cardiovascular risk factors considered included age, sex, blood pressure, body mass index, fast blood glucose, lipid profile, cardiovascular comorbidities, C reactive protein, and erythrocyte sedimentation rate. The Framingham CV risk score was performed. The sample was classified into euthyroid (n = 15), clinical hypothyroidism (n = 9), and subclinical hypothyroidism (n = 13), and included the presence of antithyroid antibodies. Fisher's exact test was used to determine the association between the variables studied. Results: 100% of the sample were women; a mean age between 39-59 years old. The category with low risk was the largest (n = 30), equivalent to 78.9%; moderate risk, no patient was obtained; high risk (n = 8) constituted 21.1%. Statistical significance between age and CV risk score in patients with clinical hypothyroidism was found (p < 1), 95% CI. The glucose level in the subclinical hypothyroidism and clinical hypothyroidism had statistical significance. The presence of anti-Thyroglobulin (antiTg) was shown to be closely related to the level of CV risk in patients with subclinical hypothyroidism. Conclusion: Age, glycemia, anti-Tg, history of DM, dyslipidemia, or cerebrovascular accidents have been linked to raising the risk of developing CVD in up to 10 years depending on their thyroid profile. No evidence of a direct relationship between CV risk score and thyroid state was found in the participants of this study.

[Use of Multiple Methods to Measure Adherence to Antiretroviral Treatment in La Romana, Dominican RepublicMedição da adesão a antirretrovirais com múltiplos métodos em La Romana, República Dominicana]. / Medición de adherencia a antirretrovirales con métodos múltiples en La Romana, República Dominicana.

Objective: To develop a tool that measures levels of adherence to antiretroviral treatment (ART) in resource-poor settings, based on a combination of four methods for measuring adherence. Methods: Retrospective review of 500 medical records of people living with HIV, randomly selected from October 2017 to January 2020. Adherence to ART was measured by combining four measurement methods (coverage of prescribed ART, ART picked up at pharmacies, viral load, and self-reported adherence). Chi-squared tests were performed with p<0.05 for statistically significant differences and logistic binary regression to identify the probability of optimal and suboptimal adherence. Spearman tests were performed for correlation of categories, and Cronbach's alpha was used to measure the internal consistency of the tool. Results: We obtained 497 adherence scores. Of these, 307 (61.8%) users qualified as adherent, 141 (28.4%) as semi-adherent, and 49 (9.8%) as non-adherent. A higher probability of optimal adherence was found in groups aged 60 years and older (odds ratio [OR]: 1.6; CI95%: 0.8-3.5), with no difference between men and women (OR: 0.9; CI95%: 0.7-1.4). Spearman's test reported a relationship (r = 0.8) between viral load levels and final score, and Cronbach's alpha yielded modest internal consistency (α = 0.7). Conclusions: A tool was developed to measure adherence to ART in a resource-poor environment. The tool shows modest levels of internal consistency and a strong correlation between viral load and adherence.

Objetivo: Desenvolver uma ferramenta para medir os níveis de adesão à terapia antirretroviral (TARV) em um ambiente de poucos recursos, com base na combinação de quatro métodos de medição de adesão. Métodos: Revisão retrospectiva de 500 prontuários de pessoas que vivem com HIV, selecionadas aleatoriamente de outubro de 2017 a janeiro de 2020. A adesão à TARV foi medida pela combinação de quatro métodos (porcentagem de cobertura da TARV prescrita, coleta de TARV na farmácia, nível de carga viral e adesão autorrelatada). Foram realizados testes de qui-quadrado com P < 0,05 para diferenças estatisticamente significativas e regressão logística binária para identificar probabilidades de adesão ótima e subótima. Foram realizados testes de Spearman para a correlação de categorias e alfa de Cronbach para medir a consistência interna do instrumento. Resultados: Foram obtidos 497 índices de adesão. Entre eles, 307 (61,8%) usuários foram classificados como aderentes, 141 (28,4%) como semiaderentes e 49 (9,8%) como não aderentes. Foi encontrada maior probabilidade de adesão ótima nos grupos de 60 anos ou mais (odds ratio [OR]: 1,6; IC95%: 0,8-3,5), sem diferença entre homens e mulheres (OR: 0,9; IC95%: 0,7-1,4). O teste de Spearman constatou uma relação (r = 0,8) entre os níveis de carga viral e a pontuação final, e o teste alfa de Cronbach mostrou uma consistência interna modesta (α = 0,7). Conclusões: Foi desenvolvida uma ferramenta para medir a adesão em um ambiente de poucos recursos. A ferramenta apresenta níveis modestos de consistência interna e forte correlação de categoria entre carga viral e adesão.

Medición de adherencia a antirretrovirales con métodos múltiples en La Romana, República Dominicana

[RESUMEN]. Objetivo. El objetivo fue desarrollar una herramienta para medir los niveles de adherencia al tratamiento antirretroviral (la TARV) en un entorno de escasos recursos, a partir de la combinación de cuatro métodos de medición de adherencia. Métodos. Revisión retrospectiva de 500 expedientes médicos de personas que viven con VIH, elegidos de manera aleatoria desde octubre del 2017 hasta enero del 2020. Se midió la adherencia a la TARV combinando cuatro métodos de medición (porcentaje de cobertura de la TARV recetada, recogida de la TARV en farma cia, nivel de carga viral y autoinforme de adherencia). Se realizaron pruebas de chi al cuadrado con P <0,05 para diferencias estadísticamente significativas y regresión binaria logística para identificar probabilidades de adherencia óptima y subóptima. Realizamos pruebas de Spearman para correlación de categorías y alfa de Cronbach para medir la consistencia interna de la herramienta. Resultados. Obtuvimos 497 calificaciones de adherencia. De estas, 307 (61,8%) usuarios se calificaron como adherentes, 141(28,4%) como semiadherentes y 49 (9,8%) como no adherentes. Se encontró una mayor probabilidad de adherencia óptima en grupos de 60 años o más (odds ratio [OR]: 1,6; IC95%: 0,8-3,5) sin diferencia entre hombres y mujeres (OR: 0,9; IC95%: 0,7-1,4). La prueba de Spearman informó una relación (r = 0,8) entre los niveles de carga viral y la calificación final, y la prueba alfa de Cronbach arrojó una modesta consistencia interna (α = 0,7). Conclusiones. Se desarrolló una herramienta para medir adherencia en un entorno de escasos recursos. La herramienta presenta niveles modestos de consistencia interna y una correlación fuerte en la categoría de carga viral y adherencia.

[RESUMO]. Objetivo. Desenvolver uma ferramenta para medir os níveis de adesão à terapia antirretroviral (TARV) em um ambiente de poucos recursos, com base na combinação de quatro métodos de medição de adesão. Métodos. Revisão retrospectiva de 500 prontuários de pessoas que vivem com HIV, selecionadas aleato riamente de outubro de 2017 a janeiro de 2020. A adesão à TARV foi medida pela combinação de quatro métodos (porcentagem de cobertura da TARV prescrita, coleta de TARV na farmácia, nível de carga viral e adesão autorrelatada). Foram realizados testes de qui-quadrado com P < 0,05 para diferenças estatistica mente significativas e regressão logística binária para identificar probabilidades de adesão ótima e subótima. Foram realizados testes de Spearman para a correlação de categorias e alfa de Cronbach para medir a con sistência interna do instrumento. Resultados. Foram obtidos 497 índices de adesão. Entre eles, 307 (61,8%) usuários foram classificados como aderentes, 141 (28,4%) como semiaderentes e 49 (9,8%) como não aderentes. Foi encontrada maior probabilidade de adesão ótima nos grupos de 60 anos ou mais (odds ratio [OR]: 1,6; IC95%: 0,8-3,5), sem diferença entre homens e mulheres (OR: 0,9; IC95%: 0,7-1,4). O teste de Spearman constatou uma relação (r = 0,8) entre os níveis de carga viral e a pontuação final, e o teste alfa de Cronbach mostrou uma consistência interna modesta (α = 0,7). Conclusões. Foi desenvolvida uma ferramenta para medir a adesão em um ambiente de poucos recursos. A ferramenta apresenta níveis modestos de consistência interna e forte correlação de categoria entre carga viral e adesão.

[ABSTRACT]. Objective. To develop a tool that measures levels of adherence to antiretroviral treatment (ART) in resour ce-poor settings, based on a combination of four methods for measuring adherence. Methods. Retrospective review of 500 medical records of people living with HIV, randomly selected from October 2017 to January 2020. Adherence to ART was measured by combining four measurement methods (coverage of prescribed ART, ART picked up at pharmacies, viral load, and self-reported adherence). Chi squared tests were performed with p<0.05 for statistically significant differences and logistic binary regression to identify the probability of optimal and suboptimal adherence. Spearman tests were performed for correlation of categories, and Cronbach's alpha was used to measure the internal consistency of the tool. Results. We obtained 497 adherence scores. Of these, 307 (61.8%) users qualified as adherent, 141 (28.4%) as semi-adherent, and 49 (9.8%) as non-adherent. A higher probability of optimal adherence was found in groups aged 60 years and older (odds ratio [OR]: 1.6; CI95%: 0.8–3.5), with no difference between men and women (OR: 0.9; CI95%: 0.7–1.4). Spearman's test reported a relationship (r = 0.8) between viral load levels and final score, and Cronbach's alpha yielded modest internal consistency (α = 0.7). Conclusions. A tool was developed to measure adherence to ART in a resource-poor environment. The tool shows modest levels of internal consistency and a strong correlation between viral load and adherence.

Covid-19, manejo obstétrico de un caso exitoso / COVID-19, obstetrical management of a successful case / COVID-19, gestão obstétrica de um caso de sucesso

RESUMEN La infección COVID-19 en su amplia manifestación de sintomas y comportamiento clínico hemodinámico en la paciente embarazada, ha demostrado que puede variar su espectro desde leve a severo. En el presente trabajo se reporta el manejo y abordaje exitoso de paciente de 24 SS de gestación con insuficiência respiratoria tipo I asociada al Sindrome de dificultad respiratoria aguda (SDRA) primario severo con fenotipo H de tipo infamatorio con hipoxemia severa y ocupación alveolar de los cuatro cuadrantes con choque séptico y neumonía severa por SARS-CoV-2 con PCR positiva para COVID-19.

ABSTRACT The COVID-19 infection, in its broad manifestation of symptoms and hemodynamic clinical behavior in the pregnant patient, has shown that its spectrum can vary from severe level. In the present work, we report the successful management and approach of a 24-SS patient with type I respiratory failure associated with severe primary Acute Respiratory Distress Syndrome (ARDS) with inflammatory-type H phenotype with severe hypoxemia and alveolar occupation quadrants with septic shock and severe pneumonia due to SARS-CoV-2 with positive PCR for COVID-19.

RESUMO A infecção pela COVID-19 em sua ampla manifestação de sintomas e comportamento clínico hemodinâmico na paciente grávida mostrou variar em espectro de leve a grave. No presente trabalho relatamos a gestão e abordagem bem sucedida de um paciente de 24 semanas de gestação com insuficiência respiratória tipo I associada à síndrome de desconforto respiratório agudo primário grave (SARS-CoV-2) com fenótipo H de tipo inflamatório com hipoxemia grave e ocupação alveolar dos quatro quadrantes com choque séptico e pneumonia grave devido à SARS-CoV-2 com PCR positiva para COVID-19.

Treatment of benign tumours and related pathologies with radiotherapy: experience of the General Hospital of Mexico.

Background: Benign diseases include tumours or localized growths with low potential for progression. The use of radiotherapy (RT) at a low dose (LD) or intermediate dose for benign pathologies has been widely proposed and studied. Currently, the use of RT is limited mainly to hyperproliferative and inflammatory diseases as a first or second line of treatment. Materials and methods: This was a retrospective, observational and descriptive study conducted in the Radiotherapy Unit of the Oncology Service of the General Hospital of Mexico "Dr. Eduardo Liceaga" from January 2, 2016, to December 31, 2020. Patients diagnosed with benign pathology and treated with RT were included. The response to treatment was recorded based on the imaging study report and/or clinical review that determined control of the disease, and toxicity was recorded based on the RTOG evaluation for acute effects and CTCAE V. 4.0 for chronic effects. Results: The records of 222 patients were analysed. The mean follow-up duration was 31.53 months (range 6-61), and the median was 24 months. Of all of the analysed pathologies that were treated with RT, keloid scars predominated in 112 patients (50.5%), and paragangliomas predominated in 72 patients (32.4%); the other patients were treated for rare pathologies. The prescribed dose was dependent on the diagnosis, with the mean dose being 31.63 Gy (1500-6000 cGy) and the median being 2000 cGy. Most of the cases of acute and chronic toxicity were grades 1 and 2, and a disease response was achieved in 94.1% of the patients. Conclusion: Our series shows that for cases of benign pathology, RT offers acceptable toxicity, improves quality of life and yields a good response, achieving disease control. These results suggest the inclusion of inflammatory pathology among the indications for treatment.

The Kinetoplastid-Specific Protein TcCAL1 Plays Different Roles During In Vitro Differentiation and Host-Cell Invasion in Trypanosoma cruzi.

In the pathogen Typanosoma cruzi, the calcium ion (Ca2+) regulates key processes for parasite survival. However, the mechanisms decoding Ca2+ signals are not fully identified or understood. Here, we investigate the role of a hypothetical Ca2+-binding protein named TcCAL1 in the in vitro life cycle of T. cruzi. Results showed that the overexpression of TcCAL1 fused to a 6X histidine tag (TcCAL1-6xHis) impaired the differentiation of epimastigotes into metacyclic trypomastigotes, significantly decreasing metacyclogenesis rates. When the virulence of transgenic metacyclic trypomastigotes was explored in mammalian cell invasion assays, we found that the percentage of infection was significantly higher in Vero cells incubated with TcCAL1-6xHis-overexpressing parasites than in controls, as well as the number of intracellular amastigotes. Additionally, the percentage of Vero cells with adhered metacyclic trypomastigotes significantly increased in samples incubated with TcCAL1-6xHis-overexpressing parasites compared with controls. In contrast, the differentiation rates from metacyclic trypomastigotes to axenic amastigotes or the epimastigote proliferation in the exponential phase of growth have not been affected by TcCAL1-6xHis overexpression. Based on our findings, we speculate that TcCAL1 exerts its function by sequestering intracellular Ca2+ by its EF-hand motifs (impairing metacyclogenesis) and/or due to an unknown activity which could be amplified by the ion binding (promoting cell invasion). This work underpins the importance of studying the kinetoplastid-specific proteins with unknown functions in pathogen parasites.

Immunization With Lipopolysaccharide-Activated Dendritic Cells Generates a Specific CD8+ T Cell Response That Confers Partial Protection Against Infection With Trypanosoma cruzi.

Lipopolysaccharide (LPS) induces the activation of dendritic cells (DCs) throughout the engagement of toll-like receptor 4. LPS-activated DCs show increased capacity to process and present pathogen-derived antigens to activate naïve T cells. DCs-based vaccines have been successfully used to treat some cancer types, and lately transferred to the field of infectious diseases, in particular against HIV. However, there is no vaccine or DC therapy for any parasitic disease that is currently available. The immune response against Trypanosoma cruzi substantially relies on T cells, and both CD4+ and CD8+ T lymphocytes are required to control parasite growth. Here, we develop a vaccination strategy based on DCs derived from bone marrow, activated with LPS and loaded with TsKb20, an immunodominant epitope of the trans-sialidase family of proteins. We extensively characterized the CD8+ T cell response generated after immunization and compared three different readouts: a tetramer staining, ELISpot and Activation-Induced Marker (AIM) assays. To our knowledge, this work shows for the first time a proper set of T cell markers to evaluate specific CD8+ T cell responses in mice. We also show that our immunization scheme confers protection against T. cruzi, augmenting survival and reducing parasite burden in female but not male mice. We conclude that the immunization with LPS-activated DCs has the potential to prime significant CD8+ T cell responses in C57BL/6 mice independently of the sex, but this response will only be effective in female, possibly due to mice sexual dimorphisms in the response generated against T. cruzi.

Malnutrición en una población de escolares en Santiago, República Dominicana / Malnutrition of an elementary school population in Santiago, Dominican Republic

Introducción: la malnutrición infantil representa uno de los problemas de salud pública más importantes de la República Dominicana (RD) y el mundo. A pesar de esto, actualmente, no existen estudios en la RD que describa el estado nutricional en los niños de la escuela primaria. Material y métodos: este estudio transversal describió las principales variables antropométricas en niños de 1ero a 6to de primaria en Santiago, RD, además de determinar la relación existente entre la antropometría y variables sociodemográficas. Resultados: de los 2,271 estudiantes estudiados, la media del peso fue 33,2 ± 11,4 kg, la talla fue 1,36 ± 0,13 m. La media del percentil fue 65,33 %. El 3.92 % (n=89) estuvo en bajo peso, el 17.57 % (n=399) estuvo en sobrepeso y el 22.94 % (n=521) estuvo en obesidad. Conclusión: el sobrepeso y la obesidad infantil fueron los trastornos más comunes en zonas rurales y urbanas, y tanto en centros privados como públicos

Introduction: Child malnutrition represents one of the most important public health issues in the Dominican Republic (DR) and the whole world. Despite this reality, there are currently no studies in the DR that describe the nutritional status in primary school children. Material and method: This cross-sectional study described the main anthropometric variables in children from elementary schools in Santiago, RD, in addition to determining the relationship between anthropometry status and some sociodemographic variables. Results: 2,271 participants were analyzed; the mean weight was 33.2 ± 11.4 kg, height was 1.36 ± 0.13 m. The mean percentile was 65.33%. 3.92% (n=89) were underweight, 17.57% (n=399) were overweight, and 22.94% (n=521) were obese. Conclusion: Childhood overweight and obesity was the most prevalent disorder, both in rural and urban areas, and both in private and public centers

Optimization and Preliminary Physicochemical Characterization of Pectin Extraction from Watermelon Rind (Citrullus lanatus) with Citric Acid.

Watermelon rind was used for the pectin extraction with citric acid as the extractant solvent. The effects of pH (2.0-3.0), extraction time (45-75 min), and liquid-solid ratio (10 : 1 to 40 : 1 mL/g) on the pectin yield, degree of esterification, methoxyl content, and anhydrouronic acid content were investigated using Box-Behnken surface response experimental design. The pH was the most significant variable for the pectin yield and properties. The responses optimized separately showed different optimal conditions for each one of the variables studied in this work. Therefore, the desirability function was used to determine the sole theoretical optimum for the highest pectin yield and highest anhydrouronic acid content, which was found to be pH of 2.0, extraction time of 62.31 min, and liquid-solid ratio of 35.07 mL/g. Under this optimal condition, the pectin yield, degree of esterification, methoxyl content, and anhydrouronic acid content were 24.30%, 73.30%, 10.45%, and 81.33%, respectively. At optimal conditions, watermelon rind pectin can be classified as high methoxyl and rapid-set pectin with high quality and high purity. Practical Applications. This study evaluated the pectin extraction from watermelon rind and carried out an optimization of multiple responses as a function of pH, time, and liquid-solid ratio to obtain the best preliminary quality parameters (pectin yield and anhydrouronic acid content). The results revealed that watermelon rind waste can be an inexpensive source to obtain good pectin quality and high purity. According to the chemical characterization and physicochemical properties studied, the extracted pectin from watermelon rind would have a high potential to be used in food industry.

Placenta accreta in early pregnancy: case report / Placenta acreta en un embarazo temprano: reporte de caso

ABSTRACT First trimester placenta accreta is an uncommon event. There are few cases reported in the literature, several associated with uterine rupture and all of them with risk factors. This pathology can be life-threatening for the mother. At such an early stage of pregnancy, abnormal placentation is diagnosed mainly due to massive hemorrhage during uterine curettage. It is of extreme importance to diagnose it before delivery or uterine evacuation, to prevent associated morbidity and mortality. We present the case of a patient with placenta accreta in early pregnancy who consulted for abnormal uterine bleeding. We describe the clinical history, imaging and management of the patient.

RESUMEN La placenta acreta del primer trimestre es un evento poco común. Hay pocos casos publicados en la literatura, varios asociados a rotura uterina y en todos ellos con factores de riesgo. Esta patología puede poner en peligro la vida de la madre. En una etapa tan temprana del embarazo, la placentación anormal se diagnostica principalmente debido a una hemorragia masiva durante el legrado uterino. Es de extrema importancia diagnosticarla antes del parto o de la evacuación uterina, para prevenir la morbimortalidad asociada. Presentamos el caso de una paciente con placenta acreta en embarazo temprano quien consultó por hemorragia uterina anormal. Describimos la historia clínica, las imágenes y el manejo realizado a la paciente.

Medición de adherencia a antirretrovirales con métodos múltiples en La Romana, República Dominicana / Use of Multiple Methods to Measure Adherence to Antiretroviral Treatment in La Romana, Dominican Republic / Medição da adesão a antirretrovirais com múltiplos métodos em La Romana, República Dominicana

RESUMEN Objetivo. El objetivo fue desarrollar una herramienta para medir los niveles de adherencia al tratamiento antirretroviral (la TARV) en un entorno de escasos recursos, a partir de la combinación de cuatro métodos de medición de adherencia. Métodos. Revisión retrospectiva de 500 expedientes médicos de personas que viven con VIH, elegidos de manera aleatoria desde octubre del 2017 hasta enero del 2020. Se midió la adherencia a la TARV combinando cuatro métodos de medición (porcentaje de cobertura de la TARV recetada, recogida de la TARV en farmacia, nivel de carga viral y autoinforme de adherencia). Se realizaron pruebas de chi al cuadrado con P <0,05 para diferencias estadísticamente significativas y regresión binaria logística para identificar probabilidades de adherencia óptima y subóptima. Realizamos pruebas de Spearman para correlación de categorías y alfa de Cronbach para medir la consistencia interna de la herramienta. Resultados. Obtuvimos 497 calificaciones de adherencia. De estas, 307 (61,8%) usuarios se calificaron como adherentes, 141(28,4%) como semiadherentes y 49 (9,8%) como no adherentes. Se encontró una mayor probabilidad de adherencia óptima en grupos de 60 años o más (odds ratio [OR]: 1,6; IC95%: 0,8-3,5) sin diferencia entre hombres y mujeres (OR: 0,9; IC95%: 0,7-1,4). La prueba de Spearman informó una relación (r = 0,8) entre los niveles de carga viral y la calificación final, y la prueba alfa de Cronbach arrojó una modesta consistencia interna (α = 0,7). Conclusiones. Se desarrolló una herramienta para medir adherencia en un entorno de escasos recursos. La herramienta presenta niveles modestos de consistencia interna y una correlación fuerte en la categoría de carga viral y adherencia.

ABSTRACT Objective. To develop a tool that measures levels of adherence to antiretroviral treatment (ART) in resource-poor settings, based on a combination of four methods for measuring adherence. Methods. Retrospective review of 500 medical records of people living with HIV, randomly selected from October 2017 to January 2020. Adherence to ART was measured by combining four measurement methods (coverage of prescribed ART, ART picked up at pharmacies, viral load, and self-reported adherence). Chi-squared tests were performed with p<0.05 for statistically significant differences and logistic binary regression to identify the probability of optimal and suboptimal adherence. Spearman tests were performed for correlation of categories, and Cronbach's alpha was used to measure the internal consistency of the tool. Results. We obtained 497 adherence scores. Of these, 307 (61.8%) users qualified as adherent, 141 (28.4%) as semi-adherent, and 49 (9.8%) as non-adherent. A higher probability of optimal adherence was found in groups aged 60 years and older (odds ratio [OR]: 1.6; CI95%: 0.8-3.5), with no difference between men and women (OR: 0.9; CI95%: 0.7-1.4). Spearman's test reported a relationship (r = 0.8) between viral load levels and final score, and Cronbach's alpha yielded modest internal consistency (α = 0.7). Conclusions. A tool was developed to measure adherence to ART in a resource-poor environment. The tool shows modest levels of internal consistency and a strong correlation between viral load and adherence.

RESUMO Objetivo. Desenvolver uma ferramenta para medir os níveis de adesão à terapia antirretroviral (TARV) em um ambiente de poucos recursos, com base na combinação de quatro métodos de medição de adesão. Métodos. Revisão retrospectiva de 500 prontuários de pessoas que vivem com HIV, selecionadas aleatoriamente de outubro de 2017 a janeiro de 2020. A adesão à TARV foi medida pela combinação de quatro métodos (porcentagem de cobertura da TARV prescrita, coleta de TARV na farmácia, nível de carga viral e adesão autorrelatada). Foram realizados testes de qui-quadrado com P < 0,05 para diferenças estatisticamente significativas e regressão logística binária para identificar probabilidades de adesão ótima e subótima. Foram realizados testes de Spearman para a correlação de categorias e alfa de Cronbach para medir a consistência interna do instrumento. Resultados. Foram obtidos 497 índices de adesão. Entre eles, 307 (61,8%) usuários foram classificados como aderentes, 141 (28,4%) como semiaderentes e 49 (9,8%) como não aderentes. Foi encontrada maior probabilidade de adesão ótima nos grupos de 60 anos ou mais (odds ratio [OR]: 1,6; IC95%: 0,8-3,5), sem diferença entre homens e mulheres (OR: 0,9; IC95%: 0,7-1,4). O teste de Spearman constatou uma relação (r = 0,8) entre os níveis de carga viral e a pontuação final, e o teste alfa de Cronbach mostrou uma consistência interna modesta (α = 0,7). Conclusões. Foi desenvolvida uma ferramenta para medir a adesão em um ambiente de poucos recursos. A ferramenta apresenta níveis modestos de consistência interna e forte correlação de categoria entre carga viral e adesão.

A Single Dose of a Hybrid hAdV5-Based Anti-COVID-19 Vaccine Induces a Long-Lasting Immune Response and Broad Coverage against VOC.

Most approved vaccines against COVID-19 have to be administered in a prime/boost regimen. We engineered a novel vaccine based on a chimeric human adenovirus 5 (hAdV5) vector. The vaccine (named CoroVaxG.3) is based on three pillars: (i) high expression of Spike to enhance its immunodominance by using a potent promoter and an mRNA stabilizer; (ii) enhanced infection of muscle and dendritic cells by replacing the fiber knob domain of hAdV5 by hAdV3; (iii) use of Spike stabilized in a prefusion conformation. The transduction with CoroVaxG.3-expressing Spike (D614G) dramatically enhanced the Spike expression in human muscle cells, monocytes and dendritic cells compared to CoroVaxG.5 that expressed the native fiber knob domain. A single dose of CoroVaxG.3 induced a potent humoral immunity with a balanced Th1/Th2 ratio and potent T-cell immunity, both lasting for at least 5 months. Sera from CoroVaxG.3-vaccinated mice was able to neutralize pseudoviruses expressing B.1 (wild type D614G), B.1.117 (alpha), P.1 (gamma) and B.1.617.2 (delta) Spikes, as well as an authentic P.1 SARS-CoV-2 isolate. Neutralizing antibodies did not wane even after 5 months, making this kind of vaccine a likely candidate to enter clinical trials.

In vitro differentiation of Trypanosoma cruzi epimastigotes into metacyclic trypomastigotes using a biphasic medium.

The pathogen Trypanosoma cruzi differentiates from epimastigotes (E) into infective metacyclic trypomastigotes (MTs) to invade the mammalian cell. This process, called metacyclogenesis, is mimicked in vitro by nutrient starvation or incubation with minimal media. Here, we describe an alternative protocol for metacyclogenesis by incubating E forms in a biphasic medium supplemented with human blood. Although time consuming, this procedure yields fully differentiated MTs without the presence of intermediate forms, even for cultures that have been maintained as E for years.

Trypanosoma cruzi Induces Regulatory B Cell Alterations in Patients With Chronic Chagas Disease.

The clinical evolution of patients with chronic Chagas disease (CCD) is mainly associated with an excessive inflammation and a defective immunomodulatory profile caused by the interaction between T. cruzi and the host. Regulatory B (Breg) cells exert immune suppression mostly through IL-10 production (B10 cells), but also through IL-10-independent mechanisms. Previously, we demonstrated that CCD patients with cardiomyopathy show changes in the ex vivo Breg cell phenotypic distribution although maintain IL-10 production capacity. Here, we sought to identify potential alterations on Breg cells upon in vitro stimulation. Isolated B cells from CCD patients with or without cardiomyopathy and non-infected (NI) donors were stimulated with T. cruzi lysate or CpG + CD40L, and characterized by flow cytometry based on the expression of CD24, CD27, CD38, and the regulatory molecules IL-10 and PD-L1. IL-10 and IL-17 secretion in the supernatant of B cells was evaluated by ELISA. Data showed that T. cruzi stimulation diminished the expression of CD24 and CD38 on CD27- B cells while reducing the percentage of CD24high inside CD27+ B cells. Furthermore, T. cruzi induced a regulatory B cell phenotype by increasing B10 cells and IL-10 secretion in all the groups. The innate-like B10 cells expansion observed in patients with cardiomyopathy would be associated with CD27- B10 cell subsets, while no predominant phenotype was found in the other groups. Patients with cardiomyopathy also displayed higher IL-17 secretion levels in T. cruzi-activated B cells. CpG + CD40L stimulation revealed that B cells from CCD patients and NI donors had the same ability to differentiate into B10 cells and secrete IL-10 in vitro. Additionally, CCD patients showed an increased frequency of CD24-CD27- B cells and a reduction in the percentage of CD24highCD27+ Breg cells, which appeared to be inversely correlated with the presence of T. cruzi DNA in blood. Finally, CCD patients exhibited a higher frequency of PD-L1+ B cells in T. cruzi-stimulated samples, suggesting that IL-10-independent mechanisms could also be tangled in the control of inflammation. Altogether, our results provide evidence about the potential role of Breg cells in the immune response developed against T. cruzi and its contribution to chronic Chagas cardiomyopathy.