Association between bacterial community and cadmium distribution across Colombian cacao crops.

Assessing the bacterial community composition across cacao crops is important to understand its potential role as a modulator of cadmium (Cd) translocation to plant tissues under field conditions; Cd mobility between soil and plants is a complex and multifactorial problem that cannot be captured only by experimentation. Although microbes have been shown to metabolize and drive the speciation of Cd under controlled conditions, regardless of the link between soil bacterial community (SBC) dynamics and Cd mobilization in the rhizosphere, only a few studies have addressed the relationship between soil bacterial community composition (SBCC) and Cd content in cacao seeds (Cdseed). Therefore, this study aimed to explore the association between SBCC and different factors influencing the distribution of Cd across cacao crop systems. This study comprised 225 samples collected across five farms, where we used an amplicon sequencing approach to characterize the bacterial community composition. The soil Cd concentration alone (Cdsoil) was a poor predictor of Cdseed. Still, we found that this relationship was more apparent when the variation within farms was controlled, suggesting a role of heterogeneity within farms in modulating Cd translocation and, thus, seed Cd content. Our results provide evidence of the link between soil bacterial communities and the distribution of Cd across Colombian cacao crops, and highlight the importance of incorporating fine-spatial-scale studies to advance the understanding of factors driving Cd uptake and accumulation in cacao plants. IMPORTANCE: Cadmium (Cd) content in cacao crops is an issue that generates interest due to the commercialization of chocolate for human consumption. Several studies provided evidence about the non-biological factors involved in its translocation into the cacao plant. However, factors related to this process, including soil bacterial community composition (SBCC), still need to be addressed. It is well known that soil microbiome could impact compounds' chemical transformation, including Cd, on the field. Here, we found the first evidence of the link between soil bacterial community composition and Cd concentration in cacao soils and seeds. It highlights the importance of including the variation of bacterial communities to assess the factors driving the Cd translocation into cacao seeds. Moreover, the results highlight the relevance of the spatial heterogeneity within and across cacao farms, influencing the variability of Cd concentrations.

Hospital-based proton therapy implementation during the COVID pandemic: early clinical and research experience in a European academic institution.

INTRODUCTION: A rapid deploy of unexpected early impact of the COVID pandemic in Spain was described in 2020. Oncology practice was revised to facilitate decision-making regarding multimodal therapy for prevalent cancer types amenable to multidisciplinary treatment in which the radiotherapy component searched more efficient options in the setting of the COVID-19 pandemic, minimizing the risks to patients whilst aiming to guarantee cancer outcomes. METHODS: A novel Proton Beam Therapy (PBT), Unit activity was analyzed in the period of March 2020 to March 2021. Institutional urgent, strict and mandatory clinical care standards for early diagnosis and treatment of COVID-19 infection were stablished in the hospital following national health-authorities' recommendations. The temporary trends of patients care and research projects proposals were registered. RESULTS: 3 out of 14 members of the professional staff involved in the PBR intra-hospital process had a positive test for COVID infection. Also, 4 out of 100 patients had positive tests before initiating PBT, and 7 out of 100 developed positive tests along the weekly mandatory special checkup performed during PBT to all patients. An update of clinical performance at the PBT Unit at CUN Madrid in the initial 500 patients treated with PBT in the period from March 2020 to November 2022 registers a distribution of 131 (26%) pediatric patients, 63 (12%) head and neck cancer and central nervous system neoplasms and 123 (24%) re-irradiation indications. In November 2022, the activity reached a plateau in terms of patients under treatment and the impact of COVID pandemic became sporadic and controlled by minor medical actions. At present, the clinical data are consistent with an academic practice prospectively (NCT05151952). Research projects and scientific production was adapted to the pandemic evolution and its influence upon professional time availability. Seven research projects based in public funding were activated in this period and preliminary data on molecular imaging guided proton therapy in brain tumors and post-irradiation patterns of blood biomarkers are reported. CONCLUSIONS: Hospital-based PBT in European academic institutions was impacted by COVID-19 pandemic, although clinical and research activities were developed and sustained. In the post-pandemic era, the benefits of online learning will shape the future of proton therapy education.

Talaromyces santanderensis: A New Cadmium-Tolerant Fungus from Cacao Soils in Colombia.

Inorganic pollutants in Colombian cocoa (Theobroma cacao L.) agrosystems cause problems in the production, quality, and exportation of this raw material worldwide. There has been an increased interest in bioprospecting studies of different fungal species focused on the biosorption of heavy metals. Furthermore, fungi constitute a valuable, profitable, ecological, and efficient natural soil resource that could be considered in the integrated management of cadmium mitigation. This study reports a new species of Talaromyces isolated from a cocoa soil sample collected in San Vicente de Chucurí, Colombia. T. santanderensis is featured by Lemon Yellow (R. Pl. IV) mycelium on CYA, mono-to-biverticillade conidiophores, and acerose phialides. T. santanderensis is distinguished from related species by its growth rate on CYAS and powdery textures on MEA, YES and OA, high acid production on CREA and smaller conidia. It is differentiated from T. lentulus by its growth rate on CYA medium at 37 °C without exudate production, its cream (R. PI. XVI) margin on MEA, and dense sporulation on YES and CYA. Phylogenetic analysis was performed using a polyphasic approach, including different phylogenetic analyses of combined and individual ITS, CaM, BenA, and RPB2 gene sequences that indicate that it is new to science and is named Talaromyces santanderensis sp. nov. This new species belongs to the Talaromyces section and is closely related to T. lentulus, T. soli, T. tumuli, and T. pratensis (inside the T. pinophilus species complex) in the inferred phylogeny. Mycelia growth of the fungal strains was subjected to a range of 0-400 mg/kg Cd and incorporated into malt extract agar (MEA) in triplicates. Fungal radial growth was recorded every three days over a 13-day incubation period and In vitro cadmium tolerance tests showed a high tolerance index (0.81) when the mycelium was exposed to 300 mg/kg of Cd. Results suggest that T. santanderensis showed tolerance to Cd concentrations that exceed the permissible limits for contaminated soils, and it is promising for its use in bioremediation strategies to eliminate Cd from highly contaminated agricultural soils.

Multi-Tissue Transcriptomic-Informed In Silico Investigation of Drugs for the Treatment of Dengue Fever Disease.

Transcriptomics, proteomics and pathogen-host interactomics data are being explored for the in silico-informed selection of drugs, prior to their functional evaluation. The effectiveness of this kind of strategy has been put to the test in the current COVID-19 pandemic, and it has been paying off, leading to a few drugs being rapidly repurposed as treatment against SARS-CoV-2 infection. Several neglected tropical diseases, for which treatment remains unavailable, would benefit from informed in silico investigations of drugs, as performed in this work for Dengue fever disease. We analyzed transcriptomic data in the key tissues of liver, spleen and blood profiles and verified that despite transcriptomic differences due to tissue specialization, the common mechanisms of action, "Adrenergic receptor antagonist", "ATPase inhibitor", "NF-kB pathway inhibitor" and "Serotonin receptor antagonist", were identified as druggable (e.g., oxprenolol, digoxin, auranofin and palonosetron, respectively) to oppose the effects of severe Dengue infection in these tissues. These are good candidates for future functional evaluation and clinical trials.

Fungal Endophytes of Tahiti Lime (Citrus citrus × latifolia) and Their Potential for Control of Colletotrichum acutatum J. H. Simmonds Causing Anthracnose.

Colletotrichum acutatum is one of the causal agents of anthracnose in several crops, and of post-flowering fruit drop (PFD) in citrus and key lime anthracnose (KLA). The pathogen normally attacks flowers, causing lesions only in open flowers. Under very favorable conditions, however, it can also affect flower buds and small fruits, causing complete rotting of the fruit and a premature fall, resulting in major economic crop losses. We isolated endophytic fungi from Tahiti lime to evaluate its diversity, verify its antagonistic capacity against the phytopathogen Colletotrichum acutatum C-100 in dual tests, and evaluate the ability of various endophytic agents to control flowers with induced anthracnose. 138 fungal isolates were obtained from 486 fragments of branches, leaves, and fruit; from which 15 species were identified morphologically. A higher isolation frequency was found in branches and leaves, with a normal level of diversity compared to other citrus species. Of the 15 morphospecies, 5 were trialed against C. acutatum in antagonism tests, resulting in a finding of positive inhibition. 2 endophytic fungi from the antagonism tests demonstrated high inhibition of the phytopathogen, and were thus used in in vivo tests with Tahiti lime flowers, applied in a spore solution. Spore solutions of two molecularly identified species, Xylaria adscendens, and Trichoderma atroviride, reduced the lesions caused by the phytopathogen in these in vivo tests. The finding that these endophytes react antagonistically against C. acutatum may make them good candidates for further biological control research in an agroindustry that requires environmental sustainability.

Association of Early Nasopharyngeal Immune Markers With COVID-19 Clinical Outcome: Predictive Value of CCL2/MCP-1.

Early recognition of severe forms of coronavirus disease 2019 (COVID-19) is essential for an opportune and effective intervention, reducing life-risking complications. An altered inflammatory immune response seems to be associated with COVID-19's pathogenesis and progression to severity. Here we demonstrate the utility of early nasopharyngeal swab samples for detection of the early expression of immune markers and the potential value of CCL2/MCP-1 in predicting disease outcome.

Caracterización de la tuberculosis en pacientes de la provincia Guantánamo, Cuba / Characterization of tuberculosis in patients from the Guantánamo province, Cuba

RESUMEN Introducción: La lucha contra la tuberculosis es una responsabilidad social y profesional que requiere de su caracterización, la que no se ha realizado en Guantánamo en la última década. Objetivo: Caracterizar la tuberculosis en pacientes de la provincia Guantánamo durante el periodo comprendido entre 2012 y 2019. Método: El universo se constituyó por el total de pacientes diagnosticados (n=136). Se estudiaron las siguientes variables: edad, sexo, localización de la enfermedad, resultados de la baciloscopía, categoría al egreso y grupos de riesgo de tuberculosis. La información se obtuvo mediante los registros de enfermedades de declaración obligatoria cada año, y las encuestas epidemiológicas de los controles de focos realizados, y se resumió en números absolutos y porcentajes. Resultados: En el 80,1 % de los casos la tuberculosis se localizó en los pulmones, y fue más común el diagnóstico de pacientes con baciloscopía positiva (63,2 %). El 76,7 % de los pacientes con baciloscopía positiva fueron masculinos. Los grupos de riesgos para tuberculosis más usuales fueron: fumadores (26,4 %), inmunodeprimidos (21,6 %) y el alcoholismo (19,1 %). Conclusiones: En la provincia Guantánamo prevalece la tuberculosis de localización pulmonar y los pacientes con bacteriología positiva. Los afectados sobre todo son hombres, tienen edad entre 45 a 54 años y son de reciente diagnóstico. La enfermedad incide más en aquellos con antecedente de ser fumadores, inmunodeprimidos y los alcohólicos.

ABSTRACT Introduction: The fight against tuberculosis is a social and professional responsibility, which requires its characterization, which has not been carried out in Guantánamo in the last decade. Objective: To characterize tuberculosis in patients in the Guantánamo province during the period between 2012 and 2019. Method: The universe was made up of the total number of diagnosed patients (n = 136). The following variables were studied: age, sex, location of the disease, smear results, category at discharge, and tuberculosis risk groups. The information was obtained through the notifiable disease registries for each year and the epidemiological surveys of the outbreak controls carried out, and was summarized in absolute numbers and percentages. Results: In 80.1% of cases, tuberculosis is in the lungs, and the diagnosis of patients with positive smear microscopy (63.2%) is more common. 76.7% of smear-positive patients are male. The most common risk groups for tuberculosis are: smokers (26.4%), immunosuppressed (21.6%) and alcoholism (19.1%). Conclusions: In Guantánamo province, pulmonary localization tuberculosis and patients with positive bacteriology prevail. Those affected are mainly men, they are between 45 and 54 years old and have recently been diagnosed. The disease affects more in those with a history of being smokers, immunosuppressed and alcoholics.

Uric Acid Has Direct Proinflammatory Effects on Human Macrophages by Increasing Proinflammatory Mediators and Bacterial Phagocytosis Probably via URAT1.

The relationship of uric acid with macrophages has not been fully elucidated. We investigated the effect of uric acid on the proinflammatory ability of human macrophages and then examined the possible molecular mechanism involved. Primary human monocytes were differentiated into macrophages for subsequent exposure to 0, 0.23, 0.45, or 0.9 mmol/L uric acid for 12 h, in the presence or absence of 1 mmol/L probenecid. Flow cytometry was used to measure proinflammatory marker production and phagocytic activity that was quantified as a percentage of GFP-labeled Escherichia coli positive macrophages. qPCR was used to measure the macrophage expression of the urate anion transporter 1 (URAT1). As compared to control cells, the production of tumor necrosis factor-alpha (TNF-alpha), toll-like receptor 4 (TLR4), and cluster of differentiation (CD) 11c was significantly increased by uric acid. In contrast, macrophages expressing CD206, CX3C-motif chemokine receptor 1 (CX3CR1), and C-C chemokine receptor type 2 (CCR2) were significantly reduced. Uric acid progressively increased macrophage phagocytic activity and downregulated URAT1 expression. Probenecid-a non-specific blocker of URAT1-dependent uric acid transport-inhibited both proinflammatory cytokine production and phagocytic activity in macrophages that were exposed to uric acid. These results suggest that uric acid has direct proinflammatory effects on macrophages possibly via URAT1.

Ontogenia e histoquímica de los esporangios y escamas receptaculares del helecho epífito Pleopeltis macrocarpa (Polypodiaceae)

Introducción: Las investigaciones sobre la ontogenia de los esporangios y más aún, de la estructura y función de las escamas receptaculares presentes en los soros de algunas especies de helechos, son escasos en la literatura científica. Objetivos: Describir y analizar la ontogenia de los esporangios y las escamas receptaculares de Pleopeltis macrocarpa. Metodología: Durante marzo y mayo de 2017 se recolectaron frondas fértiles de esta especie en los troncos de árboles en el vivero El Edén de las Flores en el municipio de Rionegro, Antioquia-Colombia. Las muestras se fijaron y procesaron de acuerdo a protocolos estándar para la inclusión y corte en parafina y resina. Las secciones obtenidas en resina (0.5 µm) se tiñeron con azul de Toluidina. Para descripciones adicionales sobre la anatomía e histoquímica se aplicaron reactivos específicos para determinar paredes primarias, secundarias, núcleos, lignina, polifenoles, polisacáridos, sustancias pécticas y celulosa. Las observaciones y registro fotográfico se efectuaron con microscopio fotónico y microscopía de epifluorescencia. Para observaciones con microscopía electrónica de barrido (MEB), los soros se deshidrataron con 2,2 dimetoxipropano, se desecaron a punto crítico y se metalizaron con oro. Resultados: Los soros son superficiales, vascularizados y de desarrollo mixto y están cubiertos por escamas receptaculares que se desprenden con la maduración de los esporangios. El esporangio de tipo leptosporangio tiene pedicelos largos de paredes primarias, anillos de los esporangios muestran paredes secundarias con engrosamientos en forma de "U" ricos en lignina. Las células epidérmicas de los receptáculos originan a los esporangios y las escamas receptaculares. Los eventos de división mitótica de estas dos estructuras son inicialmente similares, pero luego divergen para la diferenciación reproductiva y vegetativa de estos dos órganos. La meiosis es simultánea y las tétradas de esporas se disponen de forma decusada o tetragonal. El tapete celular es inicialmente uniestratificado pero por una división mitótica se torna biestratificado. Las células del estrato interno del tapete se rompen dando origen a un tapete plasmodial. En el desarrollo del esporodermo, primero se forma el exosporio, compuesto por esporopolenina, luego el endosporio compuesto de celulosa, pectina y polisacáridos carboxilados y finalmente el perisporio. Los resultados histoquímicos y de epifluorescencia indican que las paredes celulares tanto de los esporangios como las escamas receptaculares inmaduras son de naturaleza celulósica. Al madurar, estas estructuras, así como las células de la pared del esporangio mantienen esta composición. En tanto que las células epidérmicas de los escudos de las escamas receptaculares maduras se caracterizan por mostrar cutícula engrosada. Los polifenoles están presentes durante todas las etapas de desarrollo de los esporangios y escamas receptaculares. Los almidones son abundantes en etapas tempranas del desarrollo en las células del receptáculo y primordios de los esporangios. Conclusiones: La ontogenia de los esporangios de P. macrocarpa es similar al descrito para helechos leptosporangidos. Las escamas receptaculares son estructuras principalmente de protección, su morfología y composición de las paredes celulares evitan la desecación o perdida de humedad en los esporangios durante las etapas lábiles de su desarrollo. Estos resultados concuerdan con la función de protección atribuida a las escamas peltadas pluricelulares presentes en las estructuras vegetativas de algunas especies de helechos y angiospermas tolerantes a la sequía.

Introduction: The ontogeny of sporangia and furthermore the structure and function of the receptacle scales showed by the sori of some fern species are topics scarcely represented in the scientific literature. Objectives: To describe and analyze the ontogeny of sporangia and receptacle scales of Pleopeltis macrocarpa. Methods: During March and April of 2017, fertile fronds of P. macrocarpa were collected from tree stems located in the plant nursery "El Edén de las flores", municipality of Rionegro, Antioquia, Colombia. The samples were fixed and processed according to the standard protocols for embedding and sectioning in paraffin and resin. Sections obtained in resin (0.5 µm) were stained with Toluidine blue. The additional descriptions of the anatomy and histochemistry required specific reagents, applied for the determination of primary walls, secondary walls, nuclei, lignin, polyphenols, polysaccharides, pectic substances and cellulose. The observations and photographic records were performed by photonic and epifluorescence microscopy. For the scanning electron microscopy (SEM) technique, the sori were dehydrated with 2,2- Dimethoxypropane, dried to critical point and coated with gold. Results: The sori are superficial, vascularized and have mixed development, covered by receptacle scales that detach as the sporangia reaches maturity. The leptosporangiate type sporangium have long stalks of primary walls, the annulus of the sporangia shows secondary walls with "U" shaped thickenings rich in lignin. The epidermal cells of the receptacle originate the sporangia and receptacle scales. The mitotic division events of these two structures are initially similar, but then diverge for the reproductive and vegetative differentiation of these two organs. Meiosis is simultaneous and the spore tetrads are arranged in a decussate or tetragonal shape. The cellular tapetum is initially unstratified but becomes bistratified by mitotic division. The inner layer of the tapetum cells break originating a plasmodial tapetum. During the sporoderm development, the first structure formed is the exospore, composed of sporopolenin, followed by the endospore composed of cellulose, pectin and carboxilated polysaccharides, and finally the perispore. The histochemistry and epifluorescence results indicate that both the sporangia and immature receptacle scales have cell walls of cellulosic. These structures as well as those of the sporangium wall cells maintain its composition during maturation. Whereas, the epidermal wall cells of the shields from the mature receptacle scales are characterized by thickened cuticle. The polyphenols are present during all the development stages of the sporangia and receptacle scales. Starch is abundant in the early stages of development of the receptacle cells and sporangial primordia. Conclusions: The ontogeny of the sporangia of P. macrocarpa is similar to the described for leptosporangiate ferns. The receptacle scales are mainly protective structures, its morphology and cell wall composition prevent desiccation or humidity loss of the sporangia during the labile stages of development. These results agree with the protective function attributed to the peltated pluricellular scales present in the vegetative structures of drought tolerant species of ferns and angiosperms.

OSBPL10, RXRA and lipid metabolism confer African-ancestry protection against dengue haemorrhagic fever in admixed Cubans.

Ethnic groups can display differential genetic susceptibility to infectious diseases. The arthropod-born viral dengue disease is one such disease, with empirical and limited genetic evidence showing that African ancestry may be protective against the haemorrhagic phenotype. Global ancestry analysis based on high-throughput genotyping in admixed populations can be used to test this hypothesis, while admixture mapping can map candidate protective genes. A Cuban dengue fever cohort was genotyped using a 2.5 million SNP chip. Global ancestry was ascertained through ADMIXTURE and used in a fine-matched corrected association study, while local ancestry was inferred by the RFMix algorithm. The expression of candidate genes was evaluated by RT-PCR in a Cuban dengue patient cohort and gene set enrichment analysis was performed in a Thai dengue transcriptome. OSBPL10 and RXRA candidate genes were identified, with most significant SNPs placed in inferred weak enhancers, promoters and lncRNAs. OSBPL10 had significantly lower expression in Africans than Europeans, while for RXRA several SNPs may differentially regulate its transcription between Africans and Europeans. Their expression was confirmed to change through dengue disease progression in Cuban patients and to vary with disease severity in a Thai transcriptome dataset. These genes interact in the LXR/RXR activation pathway that integrates lipid metabolism and immune functions, being a key player in dengue virus entrance into cells, its replication therein and in cytokine production. Knockdown of OSBPL10 expression in THP-1 cells by two shRNAs followed by DENV2 infection tests led to a significant reduction in DENV replication, being a direct functional proof that the lower OSBPL10 expression profile in Africans protects this ancestry against dengue disease.

Control de la vía aérea en pacientes con traumatismo sobre columna cervical y clínica neurológica. Informe de casos / Airway management in patients with cervical spine trauma and neurological symptoms. Case reports

Patients with traumatic injuries of the cervical spine who undergo a surgical intervention are a great challenge to the anesthesiologist. The airway management inherently involves movements of the cervical spine that may aggravate pre-existing injury. Currently there is not a consensus for the technique of intubation of these patients. We present three patients with traumatic injuries associated neurological symptoms and cervical spine. All intubations were assisted with fiber bronchoscope, without removing the cervical collar and patients remained awake. All patients were examined again after intubation with no evidence of worsening of their neurological symptoms. We can conclude that the fiberoptic assisted intubation has several advantages in the care of these patients.

Los pacientes con lesiones traumáticas de la columna cervical que se someten a un control quirúrgico de las mismas suponen un gran reto para el anestesiólogo. El manejo de la vía aérea conlleva de forma inherente movimientos de la columna cervical que podrían agravar la lesión preexistente. Actualmente no existe un claro consenso en la técnica de intubación de estos pacientes. Presentamos tres pacientes con lesiones traumáticas de columna cervical y clínica neurológica asociada en los que se indicó cirugía. En todos ellos se realizó una intubación asistida con fibrobroncoscopio con el paciente despierto y sin retirar el collarín cervical. Todos los pacientes fueron nuevamente explorados tras la intubación sin evidenciarse en ninguno de ellos agravamiento de sus síntomas neurológicos. Podemos concluir que la intubación asistida con fibrobroncoscopio presenta varias ventajas en el cuidado de estos pacientes.

Role of CC chemokine receptor 1 and two of its ligands in human dengue infection. Three approaches under the Cuban situation.

Any of the four dengue serotypes can cause a severe disease, partly due to systemic inflammation orchestrated by mediators like cytokines and chemokines. We addressed the role of CCR1 and its ligands CCL3/MIP-1α and CCL5/RANTES in dengue infection using three different approaches: an ex vivo model exploring memory immune response in subjects with a well characterized dengue immune background, an in vivo study in patients with primary or secondary dengue infection, and an approach in fatal dengue. CCR1 and CCL3/MIP-1α gene expression showed differences after homotypic and heterotypic challenge according to dengue immune background of subjects, in correspondence with previous observations in Cuban dengue outbreaks. CCL5/RANTES gene expression was higher after homotypic challenge. CCR1 and CCL3/MIP-1α gene expression was higher in patients with secondary infection during critical days of the dengue disease, while the increase in RANTES expression started earlier than the observed for CCR1 and CCL3/MIP-1α. CCR1 and CCL3/MIP-1α gene expression was as high in brain as in spleen tissue from necropsy. Our results confirm the strong influence of previous immunity in subsequent dengue infections, and confer a possible pathogenic role to CCR1 and CCL3/MIP-1α in dengue disease and a possible protective role for CCL5/RANTES, probably through CCR5 interaction.

Variation in inflammatory/regulatory cytokines in secondary, tertiary, and quaternary challenges with dengue virus.

Secondary heterologous dengue infection is a risk factor for severe disease manifestations because of the immune-enhancement phenomenon. Succeeding clinical infections are seldom reported, and the clinical course of tertiary and quaternary dengue infections is not clear. Cuba represents a unique environment to study tertiary/quaternary dengue infections in a population with known clinical and serologic dengue markers and no dengue endemicity. We took advantage of this exceptional epidemiologic condition to study the effect of primary, secondary, tertiary, and quaternary dengue infection exposure on the expression of pro-inflammatory and regulatory cytokines, critical in dengue infection pathogenesis, by using a dengue infection ex vivo model. Whereas secondary exposure induced a high cytokine response, we found a significantly lower expression of tumor necrosis factor-α, interferon-γ, interleukin-10, and tumor growth factor-ß after tertiary and quaternary infectious challenge. Significant differences in expression of the cytokines were seen between the dengue immune profiles, suggesting that the sequence in which the immune system encounters serotypes may be important in determining the nature of the immune response to subsequent infections.

[Low-affinity Fcγ receptor IIa gene polymorphism and dengue bleeding disorder]. / Polimorfismo del receptor FcγRIIa y su posible relación con las manifestaciones clínicas del dengue.

INTRODUCTION: The pathophysiological changes that determine the severity of dengue are still not well known, therefore it is important to study the probable relationship with the host genetic. METHODS: We analyzed the possible association between the FcγRIIa polymorphism and clinical signs in individuals who suffered dengue infection in 2006, using contingency tables. RESULTS: We found that bleeding was significantly associated to FcγRIIa H/H131 genotype (80%). CONCLUSION: Our results suggest that in clinical dengue infection the bleeding could be associated to FcγRIIa H/H131 genotype.

Association of MICA and MICB alleles with symptomatic dengue infection.

Dengue viruses (DV) are one of the most important arthropod-borne viral diseases in the developing world. DV can cause syndromes that are either self-limiting or severe. Allelic variants of human leukocyte antigen (HLA) genes have been demonstrated to be associated with disease susceptibility. Here we report the association of nonclassical HLA class I MICA-MICB genes with disease outcome during DV infection. A sequencing-based typing method and genotyping of MICA and MICB in a well-characterized group of Cuban individuals with dengue hemorrhagic fever (DHF), dengue fever (DF), or asymptomatic dengue infection (ADI) was performed. Statistical analysis revealed a tendency for MICA*008 and MICB*008 to associate with susceptibility to illness when symptomatic versus asymptomatic cases (odds ratio [OR] = 2.1, p(v) = 0.03, and OR = 10.4, p = 0.0096, respectively) were compared. Surprisingly, a stronger association of both allelic forms was observed for the DF patients compared with the ADI group (MICA*008, OR = 5.2, p = 0.0001; and MICB*008, OR = 13.2, p = 0.0025) rather than the severe cases. Major histocompatibility class I-related gene-related natural killer cells and/or γδ and αß T-cell activation might regulate the development of symptomatic DF and DHF.

Long-term persistence of clinical symptoms in dengue-infected persons and its association with immunological disorders.

OBJECTIVES: The acute manifestations of dengue are well known. The clinical symptoms that present during the convalescent phase of infection are less well characterized, but may be autoimmune-based. This study was undertaken to determine the prevalence of persistent clinical symptoms among individuals infected during the 2006 Cuban epidemic and to evaluate the immunological and genetic factors associated with their occurrence. METHODS: In 2008, clinical data and blood samples were collected from a random sample of adult individuals diagnosed during the 2006 epidemic with dengue fever (DF, n=68), dengue hemorrhagic fever (DHF, n=29), or an asymptomatic infection (AI, n=42). The presence of persistent symptoms was evaluated in all individuals and a psychological assessment was performed. IgG titers and the Fc receptor (FcR) were also evaluated. The following autoimmune markers were assessed in a subset (n=26) of symptomatic individuals: complement factors C3/C4, rheumatoid factor (RF), C-reactive protein (CRP), antinuclear antibodies (ANA), and immune complex (IC). RESULTS: Over half (55/97) the individuals with a prior of diagnosis of DF or DHF had persistent clinical symptoms in the 2 years following infection. The sequelae were unrelated to the initial diagnosis and were more common among women (44/55). No symptoms were reported in the AI group and all study participants had normal mental and cognitive function. Persistent clinical symptoms were associated with HH polymorphic variant (p=0.027) and high IgG titer (p=0.041). Autoimmune marker alterations were common (20/26) in the subset of symptomatic individuals evaluated. CONCLUSIONS: Clinical sequelae after recovery from an acute dengue virus infection are common in the 2 years following infection. The results obtained in this study suggest that persistent symptoms are associated with alterations in some immunological parameters and FcγRIIa gene polymorphism. This could suggest an autoimmune-based disturbance.

Tumor necrosis factor-alpha, transforming growth factor-ß1, and interleukin-10 gene polymorphisms: implication in protection or susceptibility to dengue hemorrhagic fever.

Dengue virus infection has emerged as one of the most important arthropod-borne viral diseases. Some dengue infected individuals develop the severe, life-threatening form of the disease, dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS). Host genetic factors may be relevant and may predispose some individuals to the severe illness. Human leukocyte antigen (HLA), FcγR, tumor necrosis factor (TNF)-α, and dendritic cell-specific intracellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN), among others genes have been associated with the pathogenesis of dengue. Little is known, however, about the predictive value of cytokine genotypes for the clinical outcome of dengue infection. In this study, the TNF-α, interleukin (IL)-6, interferon (IFN)-γ, IL-10 and transforming growth factor (TGF)-ß1 gene single nucleotide polymorphisms (SNP) were studied by polymerase chain reaction-sequence-specific primer in a group of individuals with the antecedent of DHF during a secondary infection in the sequence dengue 1/dengue 2. A control group was also included. TNF-α (-308) A allele and IL-10 (-1082/-819/-592) ACC/ATA haplotype were significantly associated with DHF. TNF-α (-308) GG and TGF-ß1 (c25) GG genotypes were associated with protection. Our results suggest that genetic predisposition to a high TNF-α production and a low IL-10 production seems to increase the susceptibility to DHF during a secondary dengue 2 infection, whereas TGF-ß1 high producers might be protected for developing DHF.

MCP-1 and MIP-1α expression in a model resembling early immune response to dengue.

Dengue virus has become endemic in most tropical urban areas throughout the world, and DHF has appeared concomitantly with this expansion. The intensity of dengue virus replication during the early stages of infection could determine clinical outcomes; therefore, it is important to understand the impact of dengue virus infection on the earliest immune defense against microbial infection, which also strongly regulates the adaptive immune responses. This study was aimed at evaluating the expression of the CC-chemokines MIP-1α/CCL3 and MCP-1/CCL2 in peripheral blood leukocytes using an ex vivo model resembling dengue infection in vivo, in subjects with a well characterized dengue immune background, due to the exceptional Cuban epidemiological situation in dengue. The expression of IFNγ, TNFα and IL10 was also evaluated, giving insight about the role of MCP-1 and MIP-1α in the interplay between innate and adaptive immunity. From individuals with different dengue immune background after dengue virus challenge, increased and different expression of the chemokines and cytokines studied was verified in peripheral blood mononuclear cells, thus demonstrating that the previous immunity to a dengue virus serotype has a strong influence on the early immune response after dengue re-infection.

Asymptomatic dengue infection in a Cuban population confirms the protective role of the RR variant of the FcgammaRIIa polymorphism.

The role of human Fcgamma receptors (FcgammaR) has been recognized considerably over the last years. These receptors vary in their affinity for IgG subclasses and the intracellular signals elicited by them. Allelic variants of FcgammaR genes may influence the biological phagocyte activity, accounting for an inherited pre-disposition to disease. The specific FcgammaRIIa (CD32) contains a polymorphic variant (H/R131) that has been associated to a reduced risk for developing dengue hemorrhagic fever (DHF). Here, we investigated the role of this polymorphism in a very well-characterized group of Cuban individuals with antecedents of DHF, dengue fever (DF), or subclinical dengue infection. The HH131 genotype was significantly associated with dengue disease, either DF (*P = 0.016; odds ratio = 4.425; 95% confidence interval = 1.10-20.52) or DHF (P = 0.00018; odds ratio = 10.56; 95% confidence interval = 2.33-54.64) with respect to the subclinical infection.

Secondary heterologous dengue infection risk: Disequilibrium between immune regulation and inflammation?

Increased serum levels of cytokines released by cells of the immune response have been detected in patients suffering from dengue disease. Likewise, secondary infections by a different dengue virus serotype result in a highest risk of development of the severe dengue disease. Both findings suggest that the memory immune response is one of the key players in the pathogenesis of this disease. Here we take advantage of the particular Cuban epidemiological situation in dengue to analyze a broad spectrum of cell-mediated immune response mediators at mRNA and protein level. Evidences for a regulatory immune pattern in homologous (TGF-beta, IL-10) vs. pro-inflammatory pattern (IFN-gamma, TNF-alpha) in heterologous dengue virus re-challenge were found, suggesting a possible association with the higher incidence of severe dengue cases in the latter case.