Development of an Enzyme-Linked Immunosorbent Assay (ELISA) as a tool to detect NS1 of dengue virus serotype 2 in female Aedes aegypti eggs for the surveillance of dengue fever transmission.

Dengue is a significant disease transmitted by Aedes mosquitoes in the tropics and subtropics worldwide. The disease is caused by four virus (DENV) serotypes and is transmitted to humans by female Aedes aegypti mosquito bites infected with the virus and vertically to their progeny. Current strategies to control dengue transmission focus on the vector. In this study, we describe an indirect Enzyme-Linked Immunosorbent Assay (ELISA), using a monoclonal antibody against the non-structural dengue virus protein 1 (NS1), to detect DENV2 in Ae. aegypti eggs. The assay detects NS1 in eggs homogenates with 87.5% sensitivity and 75.0% specificity and it is proposed as a tool for the routine entomovirological surveillance of DENV 2 in field mosquito populations.

Analysis of hippocampus in rats with acute brain ischemia-reperfusion injury treated with leuprolide acetate, an agonist of GnRH.

BACKGROUND: The hippocampus is highly vulnerable to damage in the brain ischemia-reperfusion injury model. Leuprolide acetate has been shown to promote neurological recovery after injury in various regions of the central nervous system. OBJECTIVE: The objective of this study was to assess the histology of the hippocampus and the expression of neuronal recovery markers, specifically the 200 kDa neurofilaments and the myelin basic protein, in rats with brain ischemia-reperfusion injury treated with leuprolide acetate. METHODS: The rats were divided into three groups: Sham, ischemia-reperfusion with saline solution, and ischemia-reperfusion treated with leuprolide acetate. Coronal brain slices were obtained and stained with hematoxylin-eosin. The histological analysis involved quantifying the number of neurons in the hippocampal regions CA1, CA3 and DG. The myelin basic protein and neurofilaments were quantified using western blot. RESULTS: The number of neurons in CA1 and DG was significantly higher in the leuprolide acetate group compared to the untreated group. Additionally, the expression of neurofilament and myelin basic protein markers was significantly increased in rats treated with leuprolide acetate compared to the untreated rats. CONCLUSIONS: Leuprolide acetate promotes the recovery of hippocampal neurons in an acute brain ischemia-reperfusion injury model. These findings suggest that leuprolide acetate could be a potential therapeutic intervention for reversing damage in hippocampal ischemic lesions.

The costs of transgenerational immune priming for homologous and heterologous infections with different serotypes of dengue virus in Aedes aegypti mosquitoes.

The immune system is a network of molecules, signaling pathways, transcription, and effector modulation that controls, mitigates, or eradicates agents that may affect the integrity of the host. In mosquitoes, the innate immune system is highly efficient at combating foreign organisms but has the capacity to tolerate vector-borne diseases. These implications lead to replication, dissemination, and ultimately the transmission of pathogenic organisms when feeding on a host. In recent years, it has been discovered that the innate immune response of mosquitoes can trigger an enhanced immunity response to the stimulus of a previously encountered pathogen. This phenomenon, called immune priming, is characterized by a molecular response that prevents the replication of viruses, parasites, or bacteria in the body. It has been documented that immune priming can be stimulated through homologous organisms or molecules, although it has also been documented that closely related pathogens can generate an enhanced immune response to a second stimulus with a related organism. However, the cost involved in this immune response has not been characterized through the transmission of the immunological experience from parents to offspring by transgenerational immune priming (TGIP) in mosquitoes. Here, we address the impact on the rates of oviposition, hatching, development, and immune response in Aedes aegypti mosquitoes, the mothers of which were stimulated with dengue virus serotypes 2 and/or 4, having found a cost of TGIP on the development time of the progeny of mothers with heterologous infections, with respect to mothers with homologous infections. Our results showed a significant effect on the sex ratio, with females being more abundant than males. We found a decrease in transcripts of the siRNA pathway in daughters of mothers who had been exposed to an immune challenge with DV. Our research demonstrates that there are costs and benefits associated with TGIP in Aedes aegypti mosquitoes exposed to DV. Specifically, priming results in a lower viral load in the offspring of mothers who have previously been infected with the virus. Although some results from tests of two dengue virus serotypes show similarities, such as the percentage of pupae emergence, there are differences in the percentage of adult emergence, indicating differences in TGIP costs even within the same virus with different serotypes. This finding has crucial implications in the context of dengue virus transmission in endemic areas where multiple serotypes circulate simultaneously.

Enfermedad gonocócica diseminada: Reporte de caso

Introducción: La infección por Neisseria gonorrhoeae (NG) es la segunda causa enfermedad de transmisión sexual (ETS), con una incidencia en aumento, altas tasas de resistencia antibiótica y con su mayor presentación a nivel genital. Su presentación con bacteriemia es menor del 3 %, con limitante en su diagnóstico por su bajo rendimiento. Por lo anterior reportamos el caso. Presentación caso: Hombre de 60 años con antecedente de diabetes mellitus, quien ingresa por cuadro poliarticular con sinovitis, asociado a un síndrome febril, sin otro hallazgo al examen físico. Se documenta bacteriemia por NG, con nexo temporal con relación sexual previa. Se indica tratamiento antibiótico según antibiograma con profilaxis a pareja. Paciente egresa sin ninguna complicación. Conclusión : La diseminación hemática por NG es una condición rara, con síntomas iniciales muy inespecíficos, con bajo rendimiento de pruebas diagnósticas en esta condición, por lo cual requiere alta sospecha clínica y cobra gran importancia una historia clínica muy detallada. Como en nuestro caso, los síntomas inespecíficos condicionaron a descartar otras condiciones hasta el aislamiento en hemocultivos lleva a diagnósticos definitivo. Es un caso con presentación clínica inusual.

Introduction: Infection by Neisseria gonorrhoeae (NG) is the second cause of sexually transmitted disease (ETS), with an increasing incidence, high rates of antibiotic resistance and with its greatest presentation at the genital level. Its presentation with bacteriemia is less than 3%, limiting its diagnosis due to its low performance. Therefore, we report the case. Case presentation: 60-year-old man with a history of diabetes mellitus, who was admitted due to polyarticular symptoms with synovitis, associated with a febrile syndrome, with no other finding on physical examination. Bacteremia due to Neisseria gonorrhoeae is documented, with a temporal link with previous sexual intercourse. Antibiotic treatment is indicated according to antibiogram with partner prophylaxis. Patient is discharged without any complications. Conclusion: Hematic dissemination by NG is a rare condition, with very non-specific initial symptoms, with low performance of diagnostic tests in this condition, which requires high clinical suspicion, and a very detailed clinical history is of great importance. As in our case, nonspecific symptoms led to ruling out other conditions until isolation in blood cultures leads to a definitive diagnosis. It is a case with unusual clinical presentation

Detection of NS1 protein from dengue virus in excreta and homogenates of wild-caught Aedes aegypti mosquitoes using monoclonal antibodies.

Dengue fever is one of the most devastating infectious diseases worldwide. Development of methods for dengue virus (DENV) detection in mosquitoes to assess prevalence as a preliminary screen for entomological surveillance in endemic regions of DENV will certainly contribute to the control of the disease. A monoclonal antibody against the NS1 (nonstructural protein 1) viral protein was generated using recombinant NS1 protein and used to detect and analyze DENV in both excreta and total homogenates from Aedes aegypti mosquitoes. Results demonstrated expression of NS1 in excreta of DENV laboratory-infected mosquitoes and homogenates from field mosquitoes infected with DENV. The immunodetection method reported here represents a first-line strategy for assessing the prevalence of DENV in mosquitoes, for entomological surveillance in endemic regions of dengue. Detection of DENV prevalence in field mosquitoes could have an impact on vector surveillance measures to interrupt dengue transmission.

Immune priming with inactive dengue virus during the larval stage of Aedes aegypti protects against the infection in adult mosquitoes.

Several studies have observed that the immune response in insects can be conserved, a phenomenon known as immune priming, which has been mostly tested in adult stages. However, it is unknown if induction of immune priming in larval stages protects against dengue virus (DENV) infections in adult mosquitoes. In this work, we primed larval instar 3rd of Aedes aegypti with inactive dengue virus, producing adult mosquitoes with i) an enhanced antiviral-immune response; ii) a reduction in the load and replication of RNA of dengue virus (DENV); iii) a decline in viral infective particles production. Adult mosquitoes previously primed during larval stages over-expressed RNA interference (RNAi) markers Argonaute-2 (AGO-2) and Dicer-2 (DCR-2). We also observed inter-individual variations of DENV infection in adult mosquitoes, indicating a heterogeneous response to DENV infection in the same mosquito strain. However, mosquitoes primed during larval stages appear to control the infection, reducing the viral load. The over-expression of interferon-like factors (VAGO) and AGO-2 in the pupa stage suggests a fast activation of antiviral mechanisms after immune priming in larvae, creating a condition in which adult mosquitoes are resistant to the pathogen in the posterior exposure.

Stability diagrams, defect tolerance, and absorption coefficients of hybrid halide semiconductors: High-throughput first-principles characterization.

On the basis of the screened 29 hybrid halide compounds from our prior study [Y. Li and K. Yang, Energy Environ. Sci. 12, 2233-2243 (2019)], here, we reported a systematic computational study of the stability diagram, defect tolerance, and optical absorption coefficients for these candidate materials using high-throughput first-principles calculations. We took two exemplary compounds, MA2SnI4 and MA3Sb2I9, as examples to show the computational process, and they are discussed in detail. This work is expected to provide a detailed guide for further experimental synthesis and characterization, with the potential to develop novel lead-free optoelectronic devices.

Permeabilización del Canal Radicular y Transporte Apical: Comparación de Tres Sistemas / Root Canal Glide Path and Apical Transportation: Three System Comparison

La mantención de la anatomía original del canal radicular incide directamente en el éxito del tratamiento endodóntico. Para ello, los sistemas rotatorios de instrumentación requieren de canales radiculares permeables. Existen distintas limas y sistemas para la permeabilización o glide path como las limas tipo K manuales de acero inoxidable y los sistemas de NiTi rotatorios PathFile y ProGlider (Dentsply, Maillefer). Así, el objetivo de esta investigación fue comparar la capacidad de distintos sistemas de permeabilización para mantener la anatomía radicular sin producir transporte apical en canales mesiales de molares inferiores humanos extraídos. Se realizó un estudio cuantitativo experimental cuya muestra estuvo conformada por 36 canales mesiales de molares mandibulares humanos extraídos. Las muestras fueron divididas aleatoriamente en tres grupos conformados por 12 canales, cada uno de los cuales fueron sometidos a distintos sistemas de permeabilización (Lima K, PathFile y ProGlider). Las muestras fueron fotografiadas antes y después de la permeabilización utilizando un microscopio con magnificación 30X. Se cuantificó el desplazamiento del canal radicular en sentido mesio - distal y buco - lingual posterior a la permeabilización. Para el análisis de los datos se utilizó el paquete estadístico InfoStat/L y se aplicó la prueba de ANOVA / Tukey. Se observó que el sistema PathFile produce mayor transporte del canal radicular en su tercio apical en la dirección mesio-distal (p=0,77) y el sistema ProGlider en la dirección buco-lingual (p=0,57). Sin embargo, estas diferencias no fueron significativas. En conclusión, los sistemas de permeabilización analizados no presentaron diferencias en cuanto a su capacidad para mantener la anatomía sin producir transporte apical.

Preservation of the original root canal anatomy has a direct influence on the success of an endodontic treatment. In order to achieve this, rotary instrumentation systems require permeable root canals. Different files and systems are used for the establishment of a glide path such as manual stainless steel K files and NiTi rotatory systems like PathFile and ProGlider (Dentsply, Maillefer). Thus, the objective of this research was to compare the ability of different systems to create a glide path and maintain root canal anatomy without producing apical transportation in mesial root canals of extracted human lower molars. A quantitative experimental study was performed with a sample of 36 mesial root canals of extracted human mandibular molars. The samples were randomly divided into three groups consisting of 12 root canals each, which were subjected to different glide path systems (K-Files, PathFile and ProGlider). Samples were photographed before and after creating glide path using a microscope with 30X magnification. The displacement of the root canal in a mesio - distal and bucco - lingual direction was quantified after creating glide path. Data was analyzed using the statistical package InfoStat / L and the ANOVA / Tukey test was applied. The PathFile system produced greater transport of the root canal in its apical third in the mesio-distal direction (p = 0.77) and the ProGlider system in the bucco-lingual direction (p = 0.57). However, these differences were not significant. In conclusion, the glide path systems analyzed do not present any differences in their ability to maintain the anatomy without producing apical transportation.

Limited Specificity in the Injury and Infection Priming against Bacteria in Aedes aegypti Mosquitoes.

Injury and infection priming has been observed in several insect groups, reported as host immune protection against contact with a pathogen caused by a previous infection with the same. However, the specific response against a pathogen has not been demonstrated in all insect species. Investigating the specific priming response in insects is important because their immune strategies probably reflect particular selective pressures exerted by different pathogens. Here, we determined whether previous infection of Aedes aegypti would enhance survival and/or lead to greater and specific AMP expression after a second exposure to the same or a distinct bacterium. Mosquitoes previously immunized with a low dose of Escherichia coli, but not Staphylococcus aureus, showed increased survival. Although the host protection herein demonstrated was not specific, each bacterium elicited differential AMP expression. These results can be explained by the susceptible-primed-infected (SPI) epidemiological model, which poses that in the evolution of memory-like responses (priming), a pivotal role is played by pathogen virulence, associated host damage, and the host capacity of pathogen recognition.

Incorrect use of metered-dose inhalers in adult patients at a hospital in Callao, Peru, 2014: cross-sectional study.

BACKGROUND: Inhalation therapy has proven to be the best way to control the asthma and chronic obstructive pulmonary disease symptoms. The most commonly used delivery system to control these symptoms is the metered-dose inhaler. The primary goal of this study is to demonstrate an association between incorrect inhaler use and patient age. METHODS: This is a cross-sectional study, performed at Centro Médico Naval “Cirujano Mayor Santiago Távara”, in Callao, Peru, in 2014. Patients older than 18 years that used metered-dose inhalers were included. We used film recordings of patients using a metered-dose inhaler and compared their technique with the recommendations on the guidelines on the correct use of inhalers of the Spanish Society of Pneumology and Thoracic Surgery (SEPAR). The main variables measured were age and incorrect inhaler use. The results were analyzed with the Chi squared test for bivariate analysis, and for multivariate analysis we used the Poisson regression model with robust variance. RESULTS: We included 378 patients in the analysis; 167 were older than 60 years. An association was found between incorrect inhalator technique and age (p=0.014) (PR 1.19 95% CI 1.03 to 1.37). The highest prevalence of incorrect technique was found in the young adult population (88%). There was no association between the incorrect technique and the person who taught it (p=0.114). Finally, this study showed that 81.2% of the study population presented an incorrect inhalation technique. CONCLUSIONS: The percentage of incorrect inhaler use, in the general population is high. Even if we found no association between an incorrect technique and the person who taught it; still, there is a high percentage of errors and it was even demonstrated that being instructed by a pulmonologist does not guarantee a correct performance of metered-dose inhaler inhalations.

INTRODUCCIÓN : La terapia inhalatoria ha demostrado ser la más rápida y eficaz para el control del asma y la enfermedad pulmonar obstructiva crónica. El inhalador de dosis medida es el más usado por la población. El objetivo de este estudio es evidenciar la asociación entre la técnica inhalatoria incorrecta y la edad. MÉTODOS: Estudio observacional, analítico, de corte transversal realizado en Perú durante 2014. Se incluyeron pacientes desde los 18 años que utilizaran inhalador de dosis medida. Se utilizó una lista de verificación de pasos establecidos por la Sociedad Española de Neumología y Cirugía Torácica y filmaciones para evaluar la técnica inhalatoria de los pacientes. Las variables principales fueron la edad y la mala técnica inhalatoria práctica. Para el análisis bivariado se utilizó la prueba Chi cuadrado y para el análisis multivariado regresión de Poisson con varianza robusta. RESULTADOS: Se incluyeron 378 pacientes; 167 fueron mayores de 60 años. El estudio reveló que el 81,2% de la población presentó una incorrecta técnica inhalatoria. Se encontró asociación entre la edad y la técnica inhalatoria incorrecta (p=0,014) (PR 1,19 con IC 95% 1,03-1,37). El grupo etario con mayor frecuencia de técnica incorrecta fue el de adultos jóvenes (88%). CONCLUSIONES: La frecuencia de uso incorrecto del inhalador en la población es alta y esta característica predomina en el grupo de adultos jóvenes. A pesar de no haber asociación entre la persona que enseña la técnica inhalatoria y el desempeño de la misma, se demostró que existe alta frecuencia de errores, incluso en aquellos pacientes instruidos por un médico especialista.

Bacterial Exposure at the Larval Stage Induced Sexual Immune Dimorphism and Priming in Adult Aedes aegypti Mosquitoes.

Gender differences in the immune response of insects are driven by natural selection for females and sexual selection for males. These natural forces entail a multitude of extrinsic and intrinsic factors involved in a genotype-environment interaction that results in sex-biased expression of the genes shared by males and females. However, little is known about how an infection at a particular ontogenetic stage may influence later stages, or how it may impact sexual immune dimorphism. Using Aedes aegypti mosquitoes, the aim of the present study was to analyze the effect of a bacterial exposure at the larval stage on adult immunity in males and females. The parameters measured were phenoloxidase activity, nitric oxide production, antimicrobial activity, and the antimicrobial peptide transcript response. As a measure of the immune response success, the persistence of injected bacteria was also evaluated. The results show that males, as well as females, were able to enhance survival in the adult stage as a result of being exposed at the larval stage, which indicates a priming effect. Moreover, there was a differential gender immune response, evidenced by higher PO activity in males as well as higher NO production and greater antimicrobial activity in females. The greater bacterial persistence in females suggests a gender-specific strategy for protection after a previous experience with an elicitor. Hence, this study provides a primary characterization of the complex and gender-specific immune response of male and female adults against a bacterial challenge in mosquitoes primed at an early ontogenetic stage.

Is genetic structure of the southern pygmy mouse Baiomys musculus (Cricetidae) related to human-induced spatial landscape heterogeneity in a tropical dry forest?

Tropical dry forests are biologically important biomes sustaining a high rate of endemic species. However, these forests are highly threatened by human activities that negatively impact them on distinct levels, including the genetic diversity. Within the framework of landscape genetics (that seeks to evaluate the relation of characteristics of the environmental matrix with population genetics), we used ISSR markers to evaluate the relationship between the alteration of tropical dry forest by human-induced activities (conserved vs. disturbed) with the genetic structure of four breeding sites of the southern pygmy mouse Baiomys musculus. Averaging among the 105 loci used, the unbiased heterozygosis per population (0.247-0.305) was statistically similar among the four sites, as well as between conserved and disturbed conditions (2-way ANOVA F ((3,16)) = 1.47, P = 0.1984). The genetic differentiation among the four breeding sites was high (F (ST) = 0.1122; 95 % CI 0.082-0.146) considering the geographical scale evaluated (ca. 4 km). Both, a factorial correspondence analysis and a model-based clustering analysis showed the existence of four genetic groups (one per breeding site). However, no association of genetic structure with disturbance conditions was revealed by these analyses. The absence of differences in mean genetic diversity and the lack of association of genetic structure with habitat transformation suggest that B. musculus has enough behavioral plasticity and enough genetic diversity to respond to environmental heterogeneity caused by human activities. We discussed conditions in which habitat transformation could indeed favor B. musculus.