Patrones de recurrencia y resistencia asociadas a la variabilidad genética de Plasmodium vivax durante la malaria asintomática en la localidad de Mazán-Iquitos

Plasmodium vivax agente etiológico de la malaria, exhibe una gran variabilidad genética durante episodios recurrentes de la enfermedad. Esta recurrencia es informada como de baja prevalencia asociada con la malaria asintomática. Así mismo los episodios recurrentes (reinfecciones o relapsos) a menudo pueden ser confundidos por resistencia a fármacos como la cloroquina. Por lo tanto el objetivo principal de este estudio fue relacionar los patrones de recurrencia y la resistencia con la variabilidad genética de P. vivax. En este estudio se evaluaron las muestras secuenciales de individuos provenientes de una región endémica del Perú (Mazán-Iquitos), diagnosticados previamente con malaria, por microscopía, durante seguimientos activos y sometidos a un régimen de tratamiento estándar con cloroquina. La genotipificación realizada en base al gen pvmsp3-α, utilizando el Nested PCR y la digestión enzimática, permitió identificar una alta variabilidad genética de P. vivax, a partir de la cual, se identificaron los patrones de recurrencia, establecidos como relapsos, a partir de estadios latentes o hipnozoitos homólogos (con haplotipos idénticos) y reinfecciones (con haplotipos diferentes). Los rangos de tiempo permitieron una identificación más precisa, observándose mayores frecuencias de relapsos por hipnozoitos homólogos antes de los 90 días post-primera evaluación y mayores frecuencias de reinfecciones después de este periodo. Así mismo las recurrencias en el primer periodo de tiempo, por haplotipos diferentes, pueden deberse también a hipnozoitos heterólogos. Complementando el estudio, el análisis de secuenciamiento del gen pvmdr1, permitió identificar SNPs, codificantes de mutaciones no sinónimas, relacionadas con resistencia a cloroquina. Estos SNPs, a través del software U-Melt (análisis in sílico), presentaron variaciones en las temperaturas de fusión. Finalmente los resultados de cuantificación relativa con qPCR Real Time no mostraron diferencias significativas en el número de copias del gen pvmdr1. Palabras clave: Cloroquina, Genotipificación, Haplotipos, Hipnozoito, Malaria Asintomática, Recurrencia, Variabilidad.--- Plasmodium vivax etiologic agent of malaria has a large genetic variability during recurrent episodes of the disease. This recurrence is reported as low prevalence associated with asymptomatic malaria. Also recurrent episodes (reinfection or relapse) can often be mistaken for drug resistance as chloroquine. Therefore the main objective of this study was to correlate the patterns of recurrence and resistance to the genetic variability of P. vivax. In this study, we evaluated the sequential samples of individuals from an endemic region of Peru (Mazán-Iquitos), previously diagnosed with malaria microscopy during active follows and subjected to a standard treatment regimen with chloroquine. Genotyping based on the pvmsp3-α gene, using Nested PCR and enzymatic digestion, identified high genetic variability of P. vivax, from which were identified recurrence patterns established as relapse, from latent stages or homologous hypnozoites (with identical haplotypes) and reinfections (with different haplotypes). The time ranges allow more accurate identification, with higher frequency of relapses by homologous hypnozoites before 90 days post-first evaluation and higher frequencies of reinfection after this period. Also recurrences in the first period of time, for different haplotypes may also be due to heterologous hypnozoites. Complementing the study, the sequencing analysis of the gene pvmdr1, identified SNPs, encoding nonsynonymous mutations related to resistance to chloroquine. These SNPs, through U-Melt software (in sílico analysis), showed variations in the melting temperatures. Finally the results of relative cuantification with Real Time qPCR no showed significant differences in copy number of the pvmdr1 gene.} Keywords: Chloroquine, Genotyping, Haplotypes, Hipnozoite, Recurrence, Asymptomatic Malaria, Variability.Tesi

Genetic variability of Plasmodium vivax and patterns of recurrence in asymptomatic malaria at Mazan, Iquitos, Peru

El Plasmodium vivax muestra una alta variabilidad genética durante episodios recurrentes de la enfermedad. Objetivos: Determinar la variabilidad genética de P. vivax y los patrones de recurrencia durante la malaria asintomática. Diseño: Estudio descriptivo analítico. Institución: Laboratorio de Investigación en Enfermedades Infecciosas-Universidad Peruana Cayetano Heredia. Población: Individuos provenientes de Mazán-Iquitos, región endémica en malaria. Intervención: Se analizó 222 individuos con dos muestras de sangre secuenciales, entre junio de 2006 y noviembre de 2008. Principales medidas de resultados: Identificación de P. vivax, genotipificación en base al gen pvmsp3-α, variabilidad genética de P. vivax y patrones de recurrencia. Resultados: Primera evaluación: Positivos a P. vivax 191/222 (86%), a P. falciparum 2/222 (0,9%), con infección mixta 21/222 (9,5%) y negativos 8/222 (3,6%). Segunda evaluación: Permanecieron positivos a P. vivax 180/191 (94,2%). La genotipificación por nested PCR y digestión enzimática mostró haplotipos policlonales en 17/180 (9,4%) y monoclonales en 163/180 (90,6%). Se observó haplotipos diferentes (reinfección) en 88/180 (48,9%) y haplotipos idénticos (relapso) en 75/180 (41,7%). Conclusiones: Existió una alta variabilidad genética de P. vivax, y los patrones de recurrencia, basados en la genotipificación, indicaron diferencias entre reinfecciones y relapsos en individuos asintomáticos para malaria en Mazán, Iquitos.Plasmodium vivax displays a high genetic variability for recurrent episodes of illness. Objectives: To determine the genetic variability of P. vivax and patterns of recurrence in asymptomatic malaria. Design: Descriptive analytical. Setting: Laboratory of Infectious Disease Research, Universidad Peruana Cayetano Heredia. Population: Individuals from Mazan, Iquitos, Peru, a malaria endemic region. Intervention: Between June 2006 and November 2008, 222 individuals were analyzed with two sequential blood samples. Main outcome measures: Identification of P. vivax, genotyping based on gene pvmsp3-α, genetic variability of P. vivax and patterns of recurrence. Results: First evaluation: Positive for P. vivax 191/222 (86%), P. falciparum 2/222 (0.9%), mixed infection 21/222 (9.5%) and negative 8/222 (3.6%). Second evaluation: 180/191 (94.2%) remained positive for P. vivax. Genotyping by nested PCR and enzymatic digestion showed polyclonal haplotypes in 17/180 (9.4%) and monoclonal in 163/180 (90.6%). Different haplotypes (reinfection) were observed in 88/180 (48.9%) and identical haplotypes (relapse) in 75/180 (41.7%). Conclusions: There was P. vivax high genetic variability and patterns of malaria recurrence based on genotyping showed differences between reinfection and relapse in asymptomatic individuals in Mazan, Iquitos

Use of a Chagas Urine Nanoparticle Test (Chunap) to Correlate with Parasitemia Levels in T. cruzi/HIV Co-infected Patients

BackgroundEarly diagnosis of reactivated Chagas disease in HIV patients could be lifesaving. In Latin America, the diagnosis is made by microscopical detection of the T. cruzi parasite in the blood; a diagnostic test that lacks sensitivity. This study evaluates if levels of T. cruzi antigens in urine, determined by Chunap (Chagas urine nanoparticle test), are correlated with parasitemia levels in T. cruzi/HIV co-infected patients.Methodology/Principal FindingsT. cruzi antigens in urine of HIV patients (N = 55: 31 T. cruzi infected and 24 T. cruzi serology negative) were concentrated using hydrogel particles and quantified by Western Blot and a calibration curve. Reactivation of Chagas disease was defined by the observation of parasites in blood by microscopy. Parasitemia levels in patients with serology positive for Chagas disease were classified as follows: High parasitemia or reactivation of Chagas disease (detectable parasitemia by microscopy), moderate parasitemia (undetectable by microscopy but detectable by qPCR), and negative parasitemia (undetectable by microscopy and qPCR). The percentage of positive results detected by Chunap was: 100% (7/7) in cases of reactivation, 91.7% (11/12) in cases of moderate parasitemia, and 41.7% (5/12) in cases of negative parasitemia. Chunap specificity was found to be 91.7%. Linear regression analysis demonstrated a direct relationship between parasitemia levels and urine T. cruzi antigen concentrations (p 105 pg was chosen to determine patients with reactivation of Chagas disease (7/7). Antigenuria levels were 36.08 times (95% CI: 7.28 to 64.88) higher in patients with CD4+ lymphocyte counts below 200/mL (p = 0.016). No significant differences were found in HIV loads and CD8+ lymphocyte counts.ConclusionChunap shows potential for early detection of Chagas reactivation. With appropriate adaptation, this diagnostic test can be used to monitor Chagas disease status in T. cruzi/HIV co-infected patients.Author SummaryReactivation of Chagas disease in people living with HIV is a serious clinical condition that is associated with high mortality. Hence, early diagnosis and treatment can be lifesaving. Although there are not well accepted criteria to identify patients at risk of reactivation, parasitemia levels are usually considered as the best predictor. Microscopy is used in Latin America for detection of parasitemia levels. However, this has low sensitivity, which usually leads to a delay in diagnosis and treatment. Quantitative PCR is used only for research proposes in endemic areas. Antigens in urine (antigenuria) are correlated with parasitemia levels in animal models, as well as in cases of congenital Chagas disease. We believe that antigenuria can also be used for prediction of parasitemia levels in T. cruzi/HIV co-infected patients. In this study, Chunap (Chagas urine nanoparticle test) was used for concentration and quantification of T. cruzi antigens in urine of T. cruzi/HIV co-infected patients. Values of more than 105 pg of T. cruzi antigens in urine were observed only in patients with reactivation of Chagas disease. This study shows that antigenuria levels are highly correlated to levels of parasitemia and can be used as a non-invasive technique for monitoring parasitemia levels in T. cruzi/HIV co-infected patients

Multiplex Real-Time PCR Assay Using TaqMan Probes for the Identification of Trypanosoma cruzi DTUs in Biological and Clinical Samples

Background: Trypanosoma cruzi has been classified into six Discrete Typing Units (DTUs), designated as TcI–TcVI. In order to effectively use this standardized nomenclature, a reproducible genotyping strategy is imperative. Several typing schemes have been developed with variable levels of complexity, selectivity and analytical sensitivity. Most of them can be only applied to cultured stocks. In this context, we aimed to develop a multiplex Real-Time PCR method to identify the six T. cruzi DTUs using TaqMan probes (MTq-PCR).Methods/Principal Findings: The MTq-PCR has been evaluated in 39 cultured stocks and 307 biological samples from vectors, reservoirs and patients from different geographical regions and transmission cycles in comparison with a multi-locus conventional PCR algorithm. The MTq-PCR was inclusive for laboratory stocks and natural isolates and sensitive for direct typing of different biological samples from vectors, reservoirs and patients with acute, congenital infection or Chagas reactivation. The first round SL-IR MTq-PCR detected 1 fg DNA/reaction tube of TcI, TcII and TcIII and 1 pg DNA/reaction tube of TcIV, TcV and TcVI reference strains. The MTq-PCR was able to characterize DTUs in 83% of triatomine and 96% of reservoir samples that had been typed by conventional PCR methods. Regarding clinical samples, 100% of those derived from acute infected patients, 62.5% from congenitally infected children and 50% from patients with clinical reactivation could be genotyped. Sensitivity for direct typing of blood samples from chronic Chagas disease patients (32.8% from asymptomatic and 22.2% from symptomatic patients) and mixed infections was lower than that of the conventional PCR algorithm.Conclusions/Significance: Typing is resolved after a single or a second round of Real-Time PCR, depending on the DTU. This format reduces carryover contamination and is amenable to quantification, automation and kit production.This work received financial support from the Ministry of Science and Technology of Argentina [PICT 2011-0207 to AGS] and the National Scientific and Technical Research Council in Argentina (CONICET) [PIP 112 2011-010-0974 to AGS]. Work related to evaluation of biological samples was partially sponsored by the Pan-American Health Organization (PAHO) [Small Grants Program PAHO-TDR]; the Drugs and Neglected Diseases Initiative (DNDi, Geneva, Switzerland), Wellcome Trust (London, United Kingdom), SANOFI-AVENTIS (Buenos Aires, Argentina) and the National Council for Science and Technology in Mexico (CONACYT) [FONSEC 161405 to JMR]

Medio ambiente, sociedad, ética, auditoría y educación. La Investigación Contable en UNIMINUTO Virtual y a Distancia: Contexto y Oportunidades.

En los últimos años la educación a distancia ha mostrado ser capaz de reducir la brecha de inequidad social, de capacitar personas en lugares remotos, de brindar la posibilidad a la comunidad discapacitada y de permitirle a quienes no cuentan con suficientes recursos económicos acceder a la educación superior, la presente investigación da cuenta de los aspectos clave asociados a la decisión de matricularse en el programa de Contaduría Pública en la modalidad distancia tradicional. Se realizó un análisis de las narrativas basado en los resultados de un instrumento aplicado a 150 estudiantes. Para el análisis de datos se empleó la herramienta SenseMaker®. Los resultados evidencian aspectos personales, sociales y culturales que posicionan la oferta de educación en la modalidad a distancia como la institución que permite resignificar la vida de las persona

Medio ambiente, sociedad, ética, auditoría y educación. La Investigación Contable en UNIMINUTO Virtual y a Distancia: Contexto y Oportunidades.

En los últimos años la educación a distancia ha mostrado ser capaz de reducir la brecha de inequidad social, de capacitar personas en lugares remotos, de brindar la posibilidad a la comunidad discapacitada y de permitirle a quienes no cuentan con suficientes recursos económicos acceder a la educación superior, la presente investigación da cuenta de los aspectos clave asociados a la decisión de matricularse en el programa de Contaduría Pública en la modalidad distancia tradicional. Se realizó un análisis de las narrativas basado en los resultados de un instrumento aplicado a 150 estudiantes. Para el análisis de datos se empleó la herramienta SenseMaker®. Los resultados evidencian aspectos personales, sociales y culturales que posicionan la oferta de educación en la modalidad a distancia como la institución que permite resignificar la vida de las persona

Impact of COVID-19 on cardiovascular testing in the United States versus the rest of the world

Objectives: This study sought to quantify and compare the decline in volumes of cardiovascular procedures between the United States and non-US institutions during the early phase of the coronavirus disease-2019 (COVID-19) pandemic. Background: The COVID-19 pandemic has disrupted the care of many non-COVID-19 illnesses. Reductions in diagnostic cardiovascular testing around the world have led to concerns over the implications of reduced testing for cardiovascular disease (CVD) morbidity and mortality. Methods: Data were submitted to the INCAPS-COVID (International Atomic Energy Agency Non-Invasive Cardiology Protocols Study of COVID-19), a multinational registry comprising 909 institutions in 108 countries (including 155 facilities in 40 U.S. states), assessing the impact of the COVID-19 pandemic on volumes of diagnostic cardiovascular procedures. Data were obtained for April 2020 and compared with volumes of baseline procedures from March 2019. We compared laboratory characteristics, practices, and procedure volumes between U.S. and non-U.S. facilities and between U.S. geographic regions and identified factors associated with volume reduction in the United States. Results: Reductions in the volumes of procedures in the United States were similar to those in non-U.S. facilities (68% vs. 63%, respectively; p = 0.237), although U.S. facilities reported greater reductions in invasive coronary angiography (69% vs. 53%, respectively; p < 0.001). Significantly more U.S. facilities reported increased use of telehealth and patient screening measures than non-U.S. facilities, such as temperature checks, symptom screenings, and COVID-19 testing. Reductions in volumes of procedures differed between U.S. regions, with larger declines observed in the Northeast (76%) and Midwest (74%) than in the South (62%) and West (44%). Prevalence of COVID-19, staff redeployments, outpatient centers, and urban centers were associated with greater reductions in volume in U.S. facilities in a multivariable analysis. Conclusions: We observed marked reductions in U.S. cardiovascular testing in the early phase of the pandemic and significant variability between U.S. regions. The association between reductions of volumes and COVID-19 prevalence in the United States highlighted the need for proactive efforts to maintain access to cardiovascular testing in areas most affected by outbreaks of COVID-19 infection

Measurement of the top quark forward-backward production asymmetry and the anomalous chromoelectric and chromomagnetic moments in pp collisions at √s = 13 TeV

Abstract The parton-level top quark (t) forward-backward asymmetry and the anomalous chromoelectric (d̂ t) and chromomagnetic (μ̂ t) moments have been measured using LHC pp collisions at a center-of-mass energy of 13 TeV, collected in the CMS detector in a data sample corresponding to an integrated luminosity of 35.9 fb−1. The linearized variable AFB(1) is used to approximate the asymmetry. Candidate t t ¯ events decaying to a muon or electron and jets in final states with low and high Lorentz boosts are selected and reconstructed using a fit of the kinematic distributions of the decay products to those expected for t t ¯ final states. The values found for the parameters are AFB(1)=0.048−0.087+0.095(stat)−0.029+0.020(syst),μ̂t=−0.024−0.009+0.013(stat)−0.011+0.016(syst), and a limit is placed on the magnitude of | d̂ t| &lt; 0.03 at 95% confidence level. [Figure not available: see fulltext.

Measurement of the W gamma Production Cross Section in Proton-Proton Collisions at root s=13 TeV and Constraints on Effective Field Theory Coefficients

A fiducial cross section for W gamma production in proton-proton collisions is measured at a center-of-mass energy of 13 TeV in 137 fb(-1) of data collected using the CMS detector at the LHC. The W -> e nu and mu nu decay modes are used in a maximum-likelihood fit to the lepton-photon invariant mass distribution to extract the combined cross section. The measured cross section is compared with theoretical expectations at next-to-leading order in quantum chromodynamics. In addition, 95% confidence level intervals are reported for anomalous triple-gauge couplings within the framework of effective field theory.Peer reviewe

Search for dark photons in Higgs boson production via vector boson fusion in proton-proton collisions at √s = 13 TeV

A search is presented for a Higgs boson that is produced via vector boson fusion and that decays to an undetected particle and an isolated photon. The search is performed by the CMS collaboration at the LHC, using a data set corresponding to an integrated luminosity of 130 fb−1, recorded at a center-of-mass energy of 13 TeV in 2016–2018. No significant excess of events above the expectation from the standard model background is found. The results are interpreted in the context of a theoretical model in which the undetected particle is a massless dark photon. An upper limit is set on the product of the cross section for production via vector boson fusion and the branching fraction for such a Higgs boson decay, as a function of the Higgs boson mass. For a Higgs boson mass of 125 GeV, assuming the standard model production rates, the observed (expected) 95% confidence level upper limit on the branching fraction is 3.5 (2.8)%. This is the first search for such decays in the vector boson fusion channel. Combination with a previous search for Higgs bosons produced in association with a Z boson results in an observed (expected) upper limit on the branching fraction of 2.9 (2.1)% at 95% confidence level