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Epigenetic dysregulation has emerged as an important etiological mechanism of neurodevelopmental disorders (NDDs). Pathogenic variation in epigenetic regulators can impair deposition of histone post-translational modifications leading to aberrant spatiotemporal gene expression during neurodevelopment. The male-specific lethal (MSL) complex is a prominent multi-subunit epigenetic regulator of gene expression and is responsible for histone 4 lysine 16 acetylation (H4K16ac). Using exome sequencing, here we identify a cohort of 25 individuals with heterozygous de novo variants in MSL complex member MSL2. MSL2 variants were associated with NDD phenotypes including global developmental delay, intellectual disability, hypotonia, and motor issues such as coordination problems, feeding difficulties, and gait disturbance. Dysmorphisms and behavioral and/or psychiatric conditions, including autism spectrum disorder, and to a lesser extent, seizures, connective tissue disease signs, sleep disturbance, vision problems, and other organ anomalies, were observed in affected individuals. As a molecular biomarker, a sensitive and specific DNA methylation episignature has been established. Induced pluripotent stem cells (iPSCs) derived from three members of our cohort exhibited reduced MSL2 levels. Remarkably, while NDD-associated variants in two other members of the MSL complex (MOF and MSL3) result in reduced H4K16ac, global H4K16ac levels are unchanged in iPSCs with MSL2 variants. Regardless, MSL2 variants altered the expression of MSL2 targets in iPSCs and upon their differentiation to early germ layers. Our study defines an MSL2-related disorder as an NDD with distinguishable clinical features, a specific blood DNA episignature, and a distinct, MSL2-specific molecular etiology compared to other MSL complex-related disorders.
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Epilepsia , Trastornos del Neurodesarrollo , Ubiquitina-Proteína Ligasas , Adolescente , Niño , Preescolar , Femenino , Humanos , Masculino , Discapacidades del Desarrollo/genética , Metilación de ADN/genética , Epigénesis Genética , Epilepsia/genética , Histonas/metabolismo , Histonas/genética , Células Madre Pluripotentes Inducidas/metabolismo , Discapacidad Intelectual/genética , Trastornos del Neurodesarrollo/genética , Fenotipo , Ubiquitina-Proteína Ligasas/metabolismoRESUMEN
Carbapenem-resistant Klebsiella pneumoniae (CR-KP) are a public health concern, causing infections with a high mortality rate, limited therapeutic options and challenging infection control strategies. In Portugal, the CR-KP rate has increased sharply, but the factors associated with this increase are poorly explored. In order to address this question, phylogenetic and resistome analysis were used to compare the draft genomes of 200 CR-KP isolates collected in 2017-2019 from five hospitals in the Lisbon region, Portugal. Most CR-KP belonged to sequence type (ST) 13 (29%), ST17 (15%), ST348 (13%), ST231 (12%) and ST147 (7%). Carbapenem resistance was conferred mostly by the presence of KPC-3 (74%) or OXA-181 (18%), which were associated with IncF/IncN and IncX plasmids, respectively. Almost all isolates were multi-drug resistant, harbouring resistance determinants to aminoglycosides, beta-lactams, trimethoprim, fosfomycin, quinolones and sulphonamides. In addition, 11% of isolates were resistant to colistin. Colonizing and infecting isolates were highly related, and most colonized patients (89%) reported a previous hospitalization. Moreover, among the 171 events of cross-dissemination identified by core genome multi-locus sequence typing data analysis (fewer than five allelic differences), 41 occurred between different hospitals and 130 occurred within the same hospital. The results suggest that CR-KP dissemination in the Lisbon region results from acquisition of carbapenemases in mobile genetic elements, influx of CR-KP into the hospitals by colonized ambulatory patients, and transmission of CR-KP within and between hospitals. Prudent use of carbapenems, patient screening at hospital entry, and improvement of infection control are needed to decrease the burden of CR-KP infection in Portugal.
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Enterobacteriaceae Resistentes a los Carbapenémicos , Genoma Bacteriano , Hospitales , Infecciones por Klebsiella , Klebsiella pneumoniae , Portugal/epidemiología , Humanos , Infecciones por Klebsiella/microbiología , Infecciones por Klebsiella/epidemiología , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/aislamiento & purificación , Klebsiella pneumoniae/clasificación , Enterobacteriaceae Resistentes a los Carbapenémicos/genética , Enterobacteriaceae Resistentes a los Carbapenémicos/efectos de los fármacos , Enterobacteriaceae Resistentes a los Carbapenémicos/aislamiento & purificación , Enterobacteriaceae Resistentes a los Carbapenémicos/clasificación , Anciano , Persona de Mediana Edad , Masculino , Antibacterianos/farmacología , Femenino , Carbapenémicos/farmacología , Anciano de 80 o más Años , Infección Hospitalaria/microbiología , Infección Hospitalaria/epidemiología , Adulto , Plásmidos/genética , Farmacorresistencia Bacteriana Múltiple/genética , Filogenia , Adulto Joven , Pruebas de Sensibilidad Microbiana , AdolescenteRESUMEN
A common basis to address the dynamics of directly transmitted infectious diseases, such as COVID-19, are compartmental (or SIR) models. SIR models typically assume homogenous population mixing, a simplification that is convenient but unrealistic. Here we validate an existing model of a scale-free fractal infection process using high-resolution data on COVID-19 spread in São Caetano, Brazil. We find that transmission can be described by a network in which each infectious individual has a small number of susceptible contacts, of the order of 2-5. This model parameter correlated tightly with physical distancing measured by mobile phone data, such that in periods of greater distancing the model recovered a lower average number of contacts, and vice versa. We show that the SIR model is a special case of our scale-free fractal process model in which the parameter that reflects population structure is set at unity, indicating homogeneous mixing. Our more general framework better explained the dynamics of COVID-19 in São Caetano, used fewer parameters than a standard SIR model and accounted for geographically localized clusters of disease. Our model requires further validation in other locations and with other directly transmitted infectious agents.
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BACKGROUND: Accurate diagnosis of TB infection (TBI) is challenging due to the lack of a gold standard. Tuberculin skin test (TST) and interferon-gamma release assay (IGRA) are currently useful in TBI diagnosis, but both have several limitations. This study aims to evaluate inter-operator variability in TST measurements and determine its impact on TBI diagnosis and treatment.METHODS: This was a retrospective analysis of patients screened for TBI using at least TST at a public outpatient clinic specialised in TB from January 2019 to August 2021. TST readings performed by five experienced nurses were compared.RESULTS: A total of 671 screenings were analysed. TST positivity rate (P < 0.001) and mean TST measurements obtained by our nurses were significantly different (P < 0.001). Concordance of TST and IGRA results was of 83.4% in the overall population (κ = 0.479). However, TST/IGRA agreement was significantly different among nurses (P = 0.003).CONCLUSION: Our analysis of TST measurements by experienced nurses shows significant differences in TST positivity rate, mean measured values and overall concordance with IGRA. This led to significant different outcomes in TBI diagnosis and subsequent treatment. TST measurement differences could potentially be more pronounced if we considered untrained operators or those with occasional reading experience.
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Tuberculosis Latente , Humanos , Ensayos de Liberación de Interferón gamma/métodos , Tuberculosis Latente/diagnóstico , Tamizaje Masivo/métodos , Estudios Retrospectivos , Prueba de Tuberculina/métodosRESUMEN
Aromatic compounds are an important class of chemicals with different industrial applications. They are usually produced by chemical synthesis from petroleum-derived feedstocks, such as toluene, xylene and benzene. However, we are now facing threats from the excessive use of fossil fuels causing environmental problems such as global warming. Furthermore, fossil resources are not infinite, and will ultimately be depleted. To cope with these problems, the sustainable production of aromatic chemicals from renewable non-food biomass is urgent. With this in mind, the search for alternative methodologies to produce aromatic compounds using low-cost and environmentally friendly processes is becoming more and more important. Microorganisms are able to produce aromatic and aromatic-derivative compounds from sugar-based carbon sources. Metabolic engineering strategies as well as bioprocess optimization enable the development of microbial cell factories capable of efficiently producing aromatic compounds. This review presents current breakthroughs in microbial production of specialty aromatic and aromatic-derivative products, providing an overview on the general strategies and methodologies applied to build microbial cell factories for the production of these compounds. We present and describe some of the current challenges and gaps that must be overcome in order to render the biotechnological production of specialty aromatic and aromatic-derivative attractive and economically feasible at industrial scale.
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Biotecnología , Medicina , Biomasa , Ingeniería Metabólica/métodos , Compuestos Orgánicos/metabolismoRESUMEN
Pantoea agglomerans es un bacilo gramnegativo reportado principalmente en heridas penetrantes, infecciones neonatales o como contaminante. La infección relevante por esta bacteria es rara. Nuestra revisión de la literatura sugirió una asociación consistente entre la infección por Pantoea agglomerans en pacientes inmunodeprimidos y una neoplasia maligna previamente diagnosticada. Los autores describen un derrame pleural paraneumónico en el que el aislamiento de Pantoea agglomerans permitió el diagnóstico de novo de cáncer de pulmón
Pantoea agglomerans is a gram-negative rod reported mainly in penetrating wounds, neonatal infections or as a contaminant. Relevant infection by this bacterium is rare. Our review of available literature suggested a consistent association between Pantoea agglomerans infection in immunocompromised patients and previously diagnosed malignancy. The authors describe a parapneumonic pleural effusion in which the isolation of Pantoea agglomerans allowed for de novo diagnosis of lung cancer
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Humanos , Masculino , Anciano , Neoplasias Pulmonares/microbiología , Adenocarcinoma/microbiología , Inmunocompetencia , Adenocarcinoma/inmunología , Neoplasias Pulmonares/inmunología , Pantoea/aislamiento & purificación , Neoplasias Pulmonares/patología , Adenocarcinoma/patología , Derrame Pleural/microbiología , Derrame Pleural/patología , Resultado Fatal , Biomarcadores de Tumor/aislamiento & purificaciónRESUMEN
BACKGROUND: Little is known about the endoscopic and histologic findings of non-esophageal eosinophilic gastrointestinal diseases (EGID). AIM: To characterize the presenting endoscopic and histologic findings in patients with eosinophilic gastritis (EG), eosinophilic gastroenteritis (EGE), and eosinophilic colitis (EC) at diagnosis and 6 months after initiating the treatment. METHODS: We conducted a retrospective cohort study at 6 US centers associated with the Consortium of Eosinophilic Gastrointestinal Researchers. Data abstracted included demographics, endoscopic findings, tissue eosinophil counts, and associated histologic findings at diagnosis and, when available, after initial treatment. RESULTS: Of 373 subjects (317 children and 56 adults), 142 had EG, 123 EGE, and 108 EC. Normal endoscopic appearance was the most common finding across all EGIDs (62% of subjects). Baseline tissue eosinophil counts were quantified in 105 (74%) EG, 36 (29%) EGE, and 80 (74%) EC subjects. The mean peak gastric eosinophil count across all sites was 87 eos/hpf for EG and 78 eos/hpf for EGE. The mean peak colonic eosinophil count for EC subjects was 76 eos/hpf (range 10-500). Of the 29% of subjects with post-treatment follow-up, most had an improvement in clinical, endoscopic, and histologic findings regardless of treatment utilized. Reductions in tissue eosinophilia correlated with improvements in clinical symptoms as well as endoscopic and histologic findings. CONCLUSIONS: In this large cohort, normal appearance was the most common endoscopic finding, emphasizing the importance of biopsy, regardless of endoscopic appearance. Decreased tissue eosinophilia was associated with improvement in symptoms, endoscopic, and histologic findings, showing that disease activity is reversible.
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Colitis/patología , Endoscopía Gastrointestinal , Enteritis/patología , Eosinofilia/patología , Eosinófilos/patología , Gastritis/patología , Adolescente , Adulto , Anciano , Biopsia , Niño , Preescolar , Estudios de Cohortes , Colon/patología , Eritema/patología , Femenino , Humanos , Lactante , Intestino Delgado/patología , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Estómago/patología , Úlcera/patología , Adulto JovenRESUMEN
Epithelial barrier dysfunction is a significant factor in many allergic diseases, including eosinophilic esophagitis (EoE). Infiltrating leukocytes and tissue adaptations increase metabolic demands and decrease oxygen availability at barrier surfaces. Understanding of how these processes impact barrier is limited, particularly in allergy. Here, we identified a regulatory axis whereby the oxygen-sensing transcription factor HIF-1α orchestrated epithelial barrier integrity, selectively controlling tight junction CLDN1 (claudin-1). Prolonged experimental hypoxia or HIF1A knockdown suppressed HIF-1α-dependent claudin-1 expression and epithelial barrier function, as documented in 3D organotypic epithelial cultures. L2-IL5OXA mice with EoE-relevant allergic inflammation displayed localized eosinophil oxygen metabolism, tissue hypoxia, and impaired claudin-1 barrier via repression of HIF-1α/claudin-1 signaling, which was restored by transgenic expression of esophageal epithelial-targeted stabilized HIF-1α. EoE patient biopsy analysis identified a repressed HIF-1α/claudin-1 axis, which was restored via pharmacologic HIF-1α stabilization ex vivo. Collectively, these studies reveal HIF-1α's critical role in maintaining barrier and highlight the HIF-1α/claudin-1 axis as a potential therapeutic target for EoE.
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Claudina-1/metabolismo , Esofagitis Eosinofílica/metabolismo , Células Epiteliales/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Transducción de Señal , Uniones Estrechas/metabolismo , Adolescente , Adulto , Animales , Línea Celular Transformada , Niño , Preescolar , Claudina-1/genética , Esofagitis Eosinofílica/genética , Esofagitis Eosinofílica/patología , Células Epiteliales/patología , Femenino , Regulación de la Expresión Génica , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Masculino , Ratones , Ratones Transgénicos , Estabilidad Proteica , Uniones Estrechas/genética , Uniones Estrechas/patologíaRESUMEN
During mammalian embryogenesis, changes in morphology and gene expression are concurrent with epigenomic reprogramming. Using human embryonic stem cells representing the preimplantation blastocyst (naive) and postimplantation epiblast (primed), our data in 2iL/I/F naive cells demonstrate that a substantial portion of known human enhancers are premarked by H3K4me1, providing an enhanced open chromatin state in naive pluripotency. The 2iL/I/F enhancer repertoire occupies 9% of the genome, three times that of primed cells, and can exist in broad chromatin domains over 50 kb. Enhancer chromatin states are largely poised. Seventy-seven percent of 2iL/I/F enhancers are decommissioned in a stepwise manner as cells become primed. While primed topologically associating domains are largely unaltered upon differentiation, naive 2iL/I/F domains expand across primed boundaries, affecting three-dimensional genome architecture. Differential topologically associating domain edges coincide with 2iL/I/F H3K4me1 enrichment. Our results suggest that naive-derived 2iL/I/F cells have a unique chromatin landscape, which may reflect early embryogenesis.
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Blastocisto/metabolismo , Cromatina/genética , Elementos de Facilitación Genéticos/genética , Estratos Germinativos/metabolismo , Células Madre Embrionarias Humanas/metabolismo , Animales , Blastocisto/citología , Diferenciación Celular/genética , Embrión de Mamíferos/citología , Embrión de Mamíferos/embriología , Embrión de Mamíferos/metabolismo , Desarrollo Embrionario/genética , Regulación del Desarrollo de la Expresión Génica , Estratos Germinativos/citología , Células Madre Embrionarias Humanas/citología , HumanosRESUMEN
Human mobility is an important driver of geographic spread of infectious pathogens. Detailed information about human movements during outbreaks are, however, difficult to obtain and may not be available during future epidemics. The Ebola virus disease (EVD) outbreak in West Africa between 2014-16 demonstrated how quickly pathogens can spread to large urban centers following one cross-species transmission event. Here we describe a flexible transmission model to test the utility of generalised human movement models in estimating EVD cases and spatial spread over the course of the outbreak. A transmission model that includes a general model of human mobility significantly improves prediction of EVD's incidence compared to models without this component. Human movement plays an important role not only to ignite the epidemic in locations previously disease free, but over the course of the entire epidemic. We also demonstrate important differences between countries in population mixing and the improved prediction attributable to movement metrics. Given their relative rareness, locally derived mobility data are unlikely to exist in advance of future epidemics or pandemics. Our findings show that transmission patterns derived from general human movement models can improve forecasts of spatio-temporal transmission patterns in places where local mobility data is unavailable.
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Ebolavirus , Fiebre Hemorrágica Ebola/epidemiología , Fiebre Hemorrágica Ebola/transmisión , Migración Humana , Modelos Biológicos , África Occidental/epidemiología , HumanosRESUMEN
A growing number of infectious pathogens are spreading among geographic regions. Some pathogens that were previously not considered to pose a general threat to human health have emerged at regional and global scales, such as Zika and Ebola Virus Disease. Other pathogens, such as yellow fever virus, were previously thought to be under control but have recently re-emerged, causing new challenges to public health organisations. A wide array of new modelling techniques, aided by increased computing capabilities, novel diagnostic tools, and the increased speed and availability of genomic sequencing allow researchers to identify new pathogens more rapidly, assess the likelihood of geographic spread, and quantify the speed of human-to-human transmission. Despite some initial successes in predicting the spread of acute viral infections, the practicalities and sustainability of such approaches will need to be evaluated in the context of public health responses.
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The yellow fever virus (YFV) epidemic in Brazil is the largest in decades. The recent discovery of YFV in Brazilian Aedes species mosquitos highlights a need to monitor the risk of reestablishment of urban YFV transmission in the Americas. We use a suite of epidemiological, spatial, and genomic approaches to characterize YFV transmission. We show that the age and sex distribution of human cases is characteristic of sylvatic transmission. Analysis of YFV cases combined with genomes generated locally reveals an early phase of sylvatic YFV transmission and spatial expansion toward previously YFV-free areas, followed by a rise in viral spillover to humans in late 2016. Our results establish a framework for monitoring YFV transmission in real time that will contribute to a global strategy to eliminate future YFV epidemics.
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Brotes de Enfermedades/prevención & control , Monitoreo Epidemiológico , Genómica/métodos , Fiebre Amarilla/prevención & control , Fiebre Amarilla/transmisión , Virus de la Fiebre Amarilla/aislamiento & purificación , Aedes/virología , Factores de Edad , Animales , Brasil/epidemiología , Brotes de Enfermedades/estadística & datos numéricos , Evolución Molecular , Humanos , Filogenia , Reacción en Cadena de la Polimerasa , Riesgo , Factores Sexuales , Análisis Espacio-Temporal , Fiebre Amarilla/epidemiología , Fiebre Amarilla/virología , Virus de la Fiebre Amarilla/clasificación , Virus de la Fiebre Amarilla/genéticaRESUMEN
The polyphenol resveratrol (3,5,4'-trihydroxystilbene) is a well-known plant secondary metabolite, commonly used as a medical ingredient and a nutritional supplement. Due to its health-promoting properties, the demand for resveratrol is expected to continue growing. This stilbene can be found in different plants, including grapes, berries (blackberries, blueberries and raspberries), peanuts and their derived food products, such as wine and juice. The commercially available resveratrol is usually extracted from plants, however this procedure has several drawbacks such as low concentration of the product of interest, seasonal variation, risk of plant diseases and product stability. Alternative production processes are being developed to enable the biotechnological production of resveratrol by genetically engineering several microbial hosts, such as Escherichia coli, Corynebacterium glutamicum, Lactococcus lactis, among others. However, these bacterial species are not able to naturally synthetize resveratrol and therefore genetic modifications have been performed. The application of emerging metabolic engineering offers new possibilities for strain and process optimization. This mini-review will discuss the recent progress on resveratrol biosynthesis in engineered bacteria, with a special focus on the metabolic engineering modifications, as well as the optimization of the production process. These strategies offer new tools to overcome the limitations and challenges for microbial production of resveratrol in industry.
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Bacterias/genética , Bacterias/metabolismo , Ingeniería Metabólica/métodos , Estilbenos/metabolismo , Proteínas Bacterianas/genética , Vías Biosintéticas/genética , Biotecnología , Regulación Bacteriana de la Expresión Génica , Ingeniería Genética , Microbiología Industrial , Malonil Coenzima A/metabolismo , Microorganismos Modificados Genéticamente , Resveratrol , Estilbenos/químicaAsunto(s)
Anomalías Congénitas/virología , Feto/anomalías , Leche Humana/virología , Enfermedades del Sistema Nervioso/virología , Virus Zika/aislamiento & purificación , Adulto , Líquidos Corporales/virología , Brasil , Anomalías Congénitas/diagnóstico por imagen , Femenino , Feto/diagnóstico por imagen , Feto/virología , Genotipo , Humanos , Recién Nacido , Madres , Enfermedades del Sistema Nervioso/diagnóstico por imagen , Filogenia , Embarazo , ARN Viral/análisis , ARN Viral/orina , Ultrasonografía , Virus Zika/genéticaRESUMEN
OBJECTIVES: Sequelae of eosinophilic esophagitis (EoE) include food impaction and esophageal stricture. Duration of inflammation is a predicted risk factor; however, complications remain unpredictable. Studies using the functional lumen imaging probe (FLIP) have demonstrated decreased distensibility of the esophagus in adult patients with EoE. As the impact of inflammation on the developing esophagus is unknown, we investigated esophageal distensibility in a pediatric cohort to determine the effect of age, ongoing inflammation, and fibrotic features on distensibility. METHODS: We conducted a prospective observational study at two tertiary pediatric institutions. Subjects underwent FLIP evaluation during endoscopy to determine distensibility of the esophagus. During stepwise distension, simultaneous intrabag pressure and 16 channels of cross-sectional areas were measured. The minimal diameter at maximal esophageal distention at an intrabag pressure of 40 mm Hg was identified. Distensibility was compared between EoE and non-EoE subjects and between clinical variables within the EoE cohort. Potential confounding variables were identified. RESULTS: Forty-four non-EoE and 88 EoE subjects aged 3-18 years were evaluated. Age positively correlated with esophageal distensibility in the non-EoE cohort, but this trend was not observed in the EoE population. Subjects with EoE had reduced distensibility even after adjusting for age. Active inflammation (eosinophils >15 eos/high-power field), histological lamina propria fibrosis, and various features of a fibrotic phenotype (stricture, food impaction, circumferential rings on endoscopy) were associated with decreased distensibility within the EoE cohort. FLIP was safe, feasible, and well tolerated. CONCLUSIONS: These findings suggest that remodeling occurs in the pediatric EoE population, warranting early diagnosis and initiation of therapy prior to the onset of disease complications.
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Esofagitis Eosinofílica/fisiopatología , Estenosis Esofágica/fisiopatología , Adolescente , Factores de Edad , Niño , Servicios de Salud del Niño , Preescolar , Estudios de Cohortes , Colorado , Esofagoscopía , Femenino , Humanos , Masculino , Pennsylvania , Estudios ProspectivosRESUMEN
Transmission of Zika virus (ZIKV) in the Americas was first confirmed in May 2015 in northeast Brazil. Brazil has had the highest number of reported ZIKV cases worldwide (more than 200,000 by 24 December 2016) and the most cases associated with microcephaly and other birth defects (2,366 confirmed by 31 December 2016). Since the initial detection of ZIKV in Brazil, more than 45 countries in the Americas have reported local ZIKV transmission, with 24 of these reporting severe ZIKV-associated disease. However, the origin and epidemic history of ZIKV in Brazil and the Americas remain poorly understood, despite the value of this information for interpreting observed trends in reported microcephaly. Here we address this issue by generating 54 complete or partial ZIKV genomes, mostly from Brazil, and reporting data generated by a mobile genomics laboratory that travelled across northeast Brazil in 2016. One sequence represents the earliest confirmed ZIKV infection in Brazil. Analyses of viral genomes with ecological and epidemiological data yield an estimate that ZIKV was present in northeast Brazil by February 2014 and is likely to have disseminated from there, nationally and internationally, before the first detection of ZIKV in the Americas. Estimated dates for the international spread of ZIKV from Brazil indicate the duration of pre-detection cryptic transmission in recipient regions. The role of northeast Brazil in the establishment of ZIKV in the Americas is further supported by geographic analysis of ZIKV transmission potential and by estimates of the basic reproduction number of the virus.
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Infección por el Virus Zika/transmisión , Infección por el Virus Zika/virología , Virus Zika/aislamiento & purificación , Américas/epidemiología , Número Básico de Reproducción , Brasil/epidemiología , Variación Genética , Genoma Viral/genética , Humanos , Microcefalia/epidemiología , Microcefalia/virología , Epidemiología Molecular , Filogeografía , Análisis Espacio-Temporal , Virus Zika/genética , Infección por el Virus Zika/epidemiologíaRESUMEN
UNLABELLED: While combined antiretroviral therapy (cART) can result in undetectable plasma viral loads, it does not eradicate HIV infection. Furthermore, HIV-infected individuals while on cART remain at an increased risk of developing serious comorbidities, such as cancer, neurological disease, and atherosclerosis, suggesting that during cART, tissue-based HIV may contribute to such pathologies. We obtained DNA and RNA env, nef, and pol sequences using single-genome sequencing from postmortem tissues of three HIV(+) cART-treated (cART(+)) individuals with undetectable viral load and metastatic cancer at death and performed time-scaled Bayesian evolutionary analyses. We used a sensitive in situ hybridization technique to visualize HIV gag-pol mRNA transcripts in cerebellum and lymph node tissues from one patient. Tissue-associated virus evolved at similar rates in cART(+) and cART-naive (cART(-)) patients. Phylogenetic trees were characterized by two distinct features: (i) branching patterns consistent with constant viral evolution and dispersal among tissues and (ii) very recently derived clades containing both DNA and RNA sequences from multiple tissues. Rapid expansion of virus near death corresponded to wide-spread metastasis. HIV RNA(+) cells clustered in cerebellum tissue but were dispersed in lymph node tissue, mirroring the evolutionary patterns observed for that patient. Activated, infiltrating macrophages were associated with HIV RNA. Our data provide evidence that tissues serve as a sanctuary for wild-type HIV during cART and suggest the importance of macrophages as an alternative reservoir and mechanism of virus spread. IMPORTANCE: Combined antiretroviral therapy (cART) reduces plasma HIV to undetectable levels; however, removal of cART results in plasma HIV rebound, thus highlighting its inability to entirely rid the body of infection. Additionally, HIV-infected individuals on cART remain at high risk of serious diseases, which suggests a contribution from residual HIV. In this study, we isolated and sequenced HIV from postmortem tissues from three HIV(+) cART(+) individuals who died with metastatic cancer and had no detectable plasma viral load. Using high-resolution evolutionary analyses, we found that tissue-based HIV continues to replicate, evolve, and migrate among tissues during cART. Furthermore, cancer onset and metastasis coincided with increased HIV expansion, suggesting a linked mechanism. HIV-expressing cells were associated with tissue macrophages, a target of HIV infection. Our results suggest the importance of tissues, and macrophages in particular, as a target for novel anti-HIV therapies.
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Antirretrovirales/uso terapéutico , Infecciones por VIH/complicaciones , Infecciones por VIH/virología , VIH/aislamiento & purificación , Neoplasias/complicaciones , Respuesta Virológica Sostenida , Carga Viral , Terapia Antirretroviral Altamente Activa , Autopsia , Cerebelo/virología , ADN Viral/genética , Variación Genética , VIH/clasificación , VIH/genética , Infecciones por VIH/tratamiento farmacológico , Hibridación in Situ , Ganglios Linfáticos/virología , Filogenia , ARN Viral/genética , Análisis de Secuencia de ADN , Homología de Secuencia , Productos del Gen env del Virus de la Inmunodeficiencia Humana/genética , Productos del Gen nef del Virus de la Inmunodeficiencia Humana/genética , Productos del Gen pol del Virus de la Inmunodeficiencia Humana/genéticaRESUMEN
AIM: This study investigated the effect of antimicrobial photo-dynamic therapy (aPDT) over Streptococcus mutans biofilm. MATERIALS AND METHODS: Eighteen (n = 18) patients were selected and one palatine device with dental blocks was used. The biofilm was treated by curcumin and Photogem® with a LED and the effect was analyzed by CFU/ml. RESULTS: Although, statistical analysis showed significant reductions for aPDT mainly with Photogem® (p = 0.02), these were low. CONCLUSION: The results suggest a low antimicrobial effect of aPDT over S. mutans biofilm. Some parameters used need to be improved. CLINICAL SIGNIFICANCE: This technique can be a promising in Dentistry.