Human Neutrophils Couple Nitric Oxide Production and Extracellular Traps Formation in Allergic Asthma.

RATIONALE: Nitric oxide (NO) is elevated in the airways and serum of allergic asthmatic patients, suggesting an important role in asthma. NO production has been widely attributed to the canonical inducible nitric oxide synthase (iNOS). Much effort has been made to inhibit this enzyme with two outcomes: no asthma improvement; and partial NO reduction, suggesting the involvement of an iNOS-independent source. OBJECTIVES: Neutrophils produce NO under inflammatory conditions and their role in asthma has been overlooked. The present study analyzes their possible role as source of NO. METHODS: Our hypothesis was tested in 99 allergic patients with intermittent bronchial asthma and 26 healthy donors. NO production by blood and sputum neutrophils in response to allergens, anti-IgE, and anti-IgE receptors Abs was assessed by Griess, flow cytometry and confocal microscopy. Extracellular traps (ETs) formation, as a possible consequence of NO production, was quantified by western blot and confocal microscopy, and reactive oxygen species by luminol-enhanced chemiluminescence. RESULTS: Among blood and sputum granulocytes from allergic asthmatic patients, only neutrophils, produce NO by an IgE-dependent mechanism. This production is independent of NOS, but dependent on a reaction between L-arginine and reactive oxygen species from NOX2. NO and ETosis are induced in parallel, and NO amplifies ETs formation, which is a key mediator in asthma. CONCLUSIONS: Our findings reveal a novel role of neutrophils as the unique allergen/IgE-dependent NO source in allergic asthma enhancing ETs formation. These results suggest that NO produced by neutrophils needs further consideration in the treatment of allergic asthma.

Associated Factors of Pneumonia in Individuals with Chronic Obstructive Pulmonary Disease (COPD) Apart from the Use of Inhaled Corticosteroids.

Inhaled corticosteroids (ICSs) are widely used in chronic obstructive pulmonary disease (COPD) and in combination with long-acting ß2 agonists (LABAs) to reduce exacerbations and improve patient lung function and quality of life. However, ICSs have been associated with an increased risk of pneumonia in individuals with COPD, although the magnitude of this risk remains unclear. Therefore, it is difficult to make informed clinical decisions that balance the benefits and adverse effects of ICSs in people with COPD. There may be other causes of pneumonia in patients with COPD, and these causes are not always considered in studies on the risks of using ICSs in COPD. We consider it very useful to clarify these aspects in assessing the influence of ICSs on the incidence of pneumonia and their role in the treatment of COPD. This issue has important implications for current practice and the evaluation and management of COPD, since COPD patients may benefit from specific ICS-based treatment strategies. Many of the potential causes of pneumonia in patients with COPD can act synergistically, so they can be included in more than one section.

Regulation and directed inhibition of ECP production by human neutrophils.

Background: Neutrophils are involved in the pathophysiology of allergic asthma, where the Eosinophil Cationic Protein (ECP) is a critical inflammatory mediator. Although ECP production is attributed to eosinophils, we reported that ECP is also present in neutrophils from allergic patients where, in contrast to eosinophils, it is produced in an IgE-dependent manner. Given the key role of ECP in asthma, we investigated the molecular mechanisms involved in ECP production as well as the effects induced by agonists and widely used clinical approaches. We also analyzed the correlation between ECP production and lung function. Methods: Neutrophils from allergic asthmatic patients were challenged with allergens, alone or in combination with cytokines, in the presence of cell-signaling inhibitors and clinical drugs. We analyzed ECP levels by ELISA and confocal microscopy. Lung function was assessed by spirometry. Results: IgE-mediated ECP release is dependent on phosphoinositide 3-kinase, the extracellular signal-regulated kinase (ERK1/2) and the production of reactive oxygen species by NADPH-oxidase. Calcineurin phosphatase and the transcription factor NFAT are also involved. ECP release is enhanced by the cytokines interleukin (IL)-5 and granulocyte macrophage-colony stimulating factor, and inhibited by interferon-γ, IL-10, clinical drugs (formoterol, tiotropium and budesonide) and allergen-specific IT. We also found an inverse correlation between asthma severity and ECP levels. Conclusions: Our results suggest the molecular pathways involved in ECP production and potential therapeutic targets. We also provide a new method to evaluate disease severity in asthmatic patients based on the quantification of in vitro ECP production by peripheral neutrophils.

Striking lineage diversity of severe acute respiratory syndrome coronavirus 2 from non-human sources.

Due to the necessity to control human-to-human spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the overwhelming majority of the generated data on this virus was solely related to the genomic characteristics of strains infecting humans; conversely, this work aimed to recover and analyze the diversity of viral genomes from non-human sources. From a set of 3595 publicly available SARS-CoV-2 genome sequences, 128 lineages were identified in non-human hosts, the majority represented by the variants of concern Delta (n = 1105, 30.7%) and Alpha (n = 466, 12.9%), followed by B.1.1.298 lineage (n = 458, 12.7%). Environment, Neovison vison, Odocoileus virginianus and Felis catus were the non-human sources with the highest number of lineages (14, 12 and 10, respectively). Phylogenomic analyses showed viral clusters from environmental sources, N. vison, O. virginianus, Panthera tigris, and Panthera leo. These clusters were collectively related to human viruses as well as all other non-human sources that were heterogeneously distributed in the phylogenetic tree. Further, the genetic details of viral genomes from bats and pangolins were independently investigated owing to their high divergence, revealing five distinct clusters. Cluster 4 exclusively included bat-sourced genomes and the SARS-CoV-2 reference strain Wuhan-01. In summary, this study identified new genetic landmarks of SARS-CoV-2 evolution. We propose potential interspecies transmission routes of SARS-CoV-2 between animals and humans, which should be considered in order to establish better pathogen surveillance and containment strategies.

Development of an Amplicon-Based Next-Generation Sequencing Protocol to Identify Leishmania Species and Other Trypanosomatids in Leishmaniasis Endemic Areas.

Trypanosomatid infections are an important public health threat affecting many low-income countries across the tropics, particularly in the Americas. Trypanosomatids can infect many vertebrate, invertebrate, and plant species and play an important role as human pathogens. Among these clinically relevant pathogens are species from the genera Leishmania and Trypanosoma. Mixed trypanosomatid infections remain a largely unexplored phenomenon. Herein, we describe the application of an amplicon-based next-generation sequencing (NGS) assay to detect and identify trypanosomatid species in mammalian reservoirs, human patients, and sand fly vectors throughout regions of Leishmania endemicity. Sixty-five samples from different departments of Colombia, including two samples from Venezuela, were analyzed: 49 samples from cutaneous leishmaniasis (CL) patients, 8 from sand flies, 2 from domestic reservoirs (Canis familiaris), and 6 from wild reservoirs (Phyllostomus hastatus). DNA from each sample served to identify the presence of trypanosomatids through conventional PCR using heat shock protein 70 (HSP70) gene as the target. PCR products underwent sequencing by Sanger sequencing and NGS, and trypanosomatid species were identified by using BLASTn against a reference database built from trypanosomatid-derived HSP70 sequences. The alpha and beta diversity indexes of amplicon sequence variants were calculated for each group. The results revealed the presence of mixed infections with more than two Leishmania species in 34% of CL samples analyzed. Trypanosoma cruzi was identified in samples from wild reservoirs, as well as in sand fly vectors. Coinfection events with three different Leishmania species were identified in domestic reservoirs. These findings depose the traditional paradigm of leishmaniasis as being a single-species-driven infection and redraw the choreography of host-pathogen interaction in the context of multiparasitism. Further research is needed to decipher how coinfections may influence disease progression. This knowledge is key to developing an integrated approach for diagnosis and treatment. IMPORTANCE Traditionally, there has been a frequent, yet incorrect assumption that phlebotomine vectors, animal reservoirs, and human hosts are susceptible to Leishmania infection by a single parasite species. However, current evidence supports that these new vector-parasite-reservoir associations lend vectors and reservoirs greater permissiveness to certain Leishmania species, thus promoting the appearance of coinfection events, particularly in disease-endemic regions. The present study describes the application of an amplicon-based next-generation sequencing (NGS) assay to detect and identify trypanosomatid species in mammalian reservoirs, human patients, and sand fly vectors from regions of endemicity for leishmaniasis. This changes our understanding of the clinical course of leishmaniasis in areas of endemicity.

Percepción de los estudiantes de Kinesiología y Fisioterapia sobre la calidad del aula virtual

RESUMEN Introducción: la pandemia de COVID-19 ha hecho que la enseñanza en instituciones de educación superior de casi todo el mundo cambié drásticamente de la modalidad física presencial a la virtual. Objetivo: conocer la percepción que tienen los estudiantes de Kinesiología y Fisioterapia de Paraguay sobre la calidad del aula virtual en el contexto de la pandemia de COVID-19. Metodología: estudio observacional, exploratorio, descriptivo de corte transverso, prospectivo, con muestreo no probabilístico, de casos consecutivos. Un instrumento en línea fue compartido a través de las redes sociales y aplicaciones de mensajería, con el fin de estudiar características sociodemográficas y socioeconómicas, así como aquellas relacionadas a la calidad educativa. Para esto último se utilizó el "Cuestionario de Evaluación de la Calidad de los cursos virtuales de Educación Social, Calidad General del Entorno y de la Metodología Didáctica" de Santoveña Casal. Resultados: 331 estudiantes participaron de la investigación. El 74,62 % fue del sexo femenino con una edad media de 23,06±4,9. El 93,35 % utilizó por primera vez el aula virtual. Respecto a la calidad del aula virtual, el 45,62 % de los participantes manifestó que se presta "algo" de atención al proceso de enseñanza-aprendizaje y el 47,43 % mencionó que es "algo" eficaz y eficiente. 32,63 % de los participantes respondió que "muy poco" se compensa la inversión económica con la calidad del curso. Conclusión: hoy en día existe un nuevo quehacer en las labores docentes, y donde elementos de la virtualidad han comenzado a utilizarse con el objetivo de no frenar los procesos de enseñanza-aprendizaje. Nuestra investigación pone de manifiesto que la mayoría de los participantes ha utilizado por primera vez un aula virtual durante esta pandemia y, además, nos permite tener una lectura preliminar sobre la calidad de la misma en estos tiempos tan complejos.

ABSTRACT Introduction: The COVID-19 pandemic has caused teaching in higher education institutions around the world to drastically change from physical to virtual mode. Objective: To know the perception of Kinesiology and Physiotherapy students in Paraguay about the quality of the virtual classroom in the context of the COVID-19 pandemic. Methodology: observational, exploratory, descriptive, cross-sectional, prospective study, with non-probability sampling, of consecutive cases. An online instrument was shared through social networks and messaging applications, in order to study sociodemographic and socioeconomic characteristics, as well as those related to educational quality. For the latter, the "Cuestionario de Evaluación de la Calidad de los cursos virtuales de Educación Social, Calidad General del Entorno y de la Metodología Didáctica" by Santoveña Casal was used. Results: 331 students participated in the research. 74.62% were female with a mean age of 23.06±4.9. 93.35% used the virtual classroom for the first time. Regarding the quality of the virtual classroom, 45.62% of the participants stated that "some" attention is paid to the teaching-learning process and 47.43% mentioned that it is "somewhat" effective and efficient. 32.63% of the participants responded that "very little" is compensated for the economic investment with the quality of the course. Conclusion: Today there is a new task in the teaching work, and where elements of virtuality have begun to be used with the aim of not slowing down the teaching-learning processes. Our research shows that most of the participants have used a virtual classroom for the first time during this pandemic and, in addition, it allows us to have a preliminary reading on the quality of it in these complex times.

SARS-CoV-2 spread across the Colombian-Venezuelan border.

INTRODUCTION: Venezuela and Colombia both adopted measures of containment early in response to the COVID-19 pandemic. However, Venezuela's ongoing humanitarian crisis has decimated its health care system, and forced millions of Venezuelans to flee through its porous border with Colombia. The extensive shared border, and illegal cross-border transit through improvised trails between the two countries are major challenges for public health authorities. We report the first SARS-CoV-2 genomes from Venezuela, and present a snapshot of the SARS-CoV-2 epidemiologic landscape in the Colombian-Venezuelan border region. METHODS: We sequenced and assembled viral genomes from total RNA extracted from nasopharyngeal (NP) clinical specimens using a custom reference-based analysis pipeline. Three assemblies obtained were subjected to typing using the Phylogenetic Assignment of Named Global Outbreak LINeages 'Pangolin' tool. A total of 376 publicly available SARS-CoV-2 genomes from South America were obtained from the GISAID database to perform comparative genomic analyses. Additionally, the Wuhan-1 strain was used as reference. RESULTS: We found that two of the SARS-CoV-2 genomes from Venezuela belonged to the B1 lineage, and the third to the B.1.13 lineage. We observed a point mutation in the Spike protein gene (D614G substitution), previously reported to be associated with increased infectivity, in all three Venezuelan genomes. Additionally, three mutations (R203K/G204R substitution) were present in the nucleocapsid (N) gene of one Venezuelan genome. CONCLUSIONS: Genomic sequencing demonstrates similarity between SARS-CoV-2 lineages from Venezuela and viruses collected from patients in bordering areas in Colombia and from Brazil, consistent with cross-border transit despite administrative measures including lockdowns. The presence of mutations associated with increased infectivity in the 3 Venezuelan genomes we report and Colombian SARS-CoV-2 genomes from neighboring borders areas may pose additional challenges for control of SARS-CoV-2 spread in the complex epidemiological landscape in Latin American countries. Public health authorities should carefully follow the progress of the pandemic and its impact on displaced populations within the region.

Allergens Induce the Release of Lactoferrin by Neutrophils from Asthmatic Patients.

BACKGROUND: Despite the evidence that Lactoferrin (Lf) is involved in allergic asthma processes, it is unknown whether neutrophils can be one of the main cellular sources of this key inflammatory mediator directly in response of an IgE mediated stimulus. The present study was undertaken to analyze this question. METHODS: Neutrophils from healthy subjects (n = 34) and neutrophils from allergic asthmatic patients (n = 102) were challenged in vitro with specific allergens to which the patients were sensitized, PAF, or agonist mAbs against IgE-receptors, and the levels of Lf were measured in the culture supernatant. The levels of serum IgE together with the severity of symptoms were also analyzed. RESULTS: Lf was released into the culture supernatant of neutrophils from allergic asthmatic patients in response to allergens and PAF. This response was highly allergen-specific, and did not happen in neutrophils from healthy donors. Allergen effect was mimicked by Abs against FcεRI and galectin-3 but not by FcεRII. The levels of released Lf correlated well with the levels of serum specific IgE and severity of asthma symptoms. These observations represent a novel view of neutrophils as an important source of Lf in allergic asthma. Importantly, the levels of released Lf by neutrophils could therefore be used to evaluate disease severity in allergic asthmatic patients.

Dual DNA vaccination of rainbow trout (Oncorhynchus mykiss) against two different rhabdoviruses, VHSV and IHNV, induces specific divalent protection.

DNA vaccines encoding the glycoprotein genes of the salmonid rhabdoviruses VHSV and IHNV are very efficient in eliciting protective immune responses against their respective diseases in rainbow trout (Oncorhynchus mykiss). The early anti-viral response (EAVR) provides protection by 4 days post vaccination and is non-specific and transient while the specific anti-viral response (SAVR) is long lasting and highly specific. Since both VHSV and IHNV are endemic in rainbow trout in several geographical regions of Europe and Atlantic salmon (Salmo salar) on the Pacific coast of North America, co-vaccination against the two diseases would be a preferable option. In the present study we demonstrated that a single injection of mixed DNA vaccines induced long-lasting protection against both individual and a simultaneous virus challenge 80 days post vaccination. Transfected muscle cells at the injection site expressed both G proteins. This study confirms the applied potential of using a combined DNA vaccination for protection of fish against two different rhabdoviral diseases.

Enfermedades de transmisión sexual en el curso del VIH/SIDA. Sanatorio Santiago de las Vegas 1997-1998 / Sexually trasnmitted diseases in the course of HIV/AIDS

Se seleccionaron 42 pacientes seropositivos al VIH/SIDA que fueron diagnosticados en el período de 1 año con otra enfermedad de transmisión sexual, teniendo en cuenta que éstas han constituido en todos los tiempos un gran problema de salud y con el objetivo de determinar el nivel de conocimientos de los pacientes acerca de estas afecciones y la eficiencia de la labor de enfermería en la actividad preventivo-educativa. Con la aplicación de encuestas se comprobó el bajo nivel de conocimientos que poseían la mayoría de los pacientes, ya que el 70 porciento demostró poseer conocimientos muy bajos, por lo que se elaboró una estrategia de intervención a través de actividades de educación para la salud, que con su puesta en marcha logró resultados alentadores y satisfactorios, ya que del porcentaje inicial se redujo al 15 porciento los pacientes con bajo nivel de conocimientos y aumentó la cifra en los niveles de conocimientos mediano y elevado hasta el 85 porciento. Además se incentivó al personal de enfermería a potenciar su labor preventivo-educativa, encomienda ésta que constituye parte medular en el desempeño de sus funciones

Enterotest y parasitismo intestinal / Enterotest and intestinal parasitism

Se comparó la sensibilidad de la cuerda encapsulada con el examen seriado de heces, para demostrar parasitismo intestinal en treinta pacientes admitidos en el Servicio de Tropa del Hospital MIlitar Central. Los resultados demostraron una alta incidencia de parasitismo intestinal en nuestros pacientes, una baja sensibilidad de la cuerda encapsulada y la elevada morbilidad que produce esta infestación: anemia crónica, diarrea y desnutrición.

Tuberculosis hepática y fiebre de origen desconocido / Hepatic tuberculosis and fever of unknown origin

Se presenta una paciente de 44 años de edad, proveniente de Iquitos con fiebre de tres meses de evolución y pérdida progresiva de peso; el examen clínico fue normal a excepción de hepatomegalia moderada. Frente a este síndrome febril crónico de origen desconocido, se programó una serie de procedimientos y exámenes auxiliares en forma gradual y sistemática; siguiendo un protocolo previamente establecido en nuestro servicio para estos casos. La biopsia laparoscópica del hígado mostró conglomerados linfomonocitarios múltiples y Mycobacterium tuberculosis. La evolución de la paciente luego del tratamiento específico fue satisfactoria, dándosele de alta (diecinueve días después de iniciarse la terapia) afebril, en buenas condiciones físicas para continuar su tratamiento en forma ambulatoria. Presentamos esta paciente con tuberculosis extrapulmonar hepática micronodular múltiple que cursa con fiebre de más de tres semanas de evolución para insistir que en nuestro medio es necesario considerar esta etiología, dada su prevalencia, como causa de síndrome febril crónico de origen desconocido en pacientes adultos con hepatomegalia.

Metaplasia mieloide agnogénica: reporte de un caso atípico / Agnogenic myeloid metaplasia: case of report atipic

Se presenta un paciente varón de 19 años de edad, procedente de Papayacu, departamento de Amazonas, con metaplasia mieloide agnogénica (MMA), cuya sintomatología y evolución-con excepción de la fiebre y la esplenomegalia-fueron inusuales, lo cual obscureció el diagnóstico que sólo pudo evidenciarse post-mortem. Paralelamente era portador de una severa desnutrición calórico-proteica y de parasitismo intestinal múltiple, lo que contribuyó a modificar el curso clínico del proceso. Creemos por ello que este caso de metaplasia mieloide agnogénica de presentación muy singular debe ser reportado en la literatura nacional.

Fiebre tifoidea y ciprofloxacina / Typhoid fever and ciprofloxacine

Se presenta a un varón, de 20 años de edad, procedente de Andahuaylas, con fiebre tifoidea complicada con hepatitis severa, ascitis, hemorragia digestiva, deshidratación e insuficiencia prerrenal aguda; tratado exitosamente con un ciclo corto de ciprofloxacina endovenosa: 400 mg bid durante seis días, en el Hospital Militar Central de Lima. Concomitantemente, el paciente sufrió de malaria y parasitismo intestinal con Ancylostoma duodenale. Al trigésimo sexto día de su internamiento, y estando asintomático, desarrollo neumonía basal izquierda que se trató con eritromicina oral. Se hace una revisión de la literatura en relación al uso de quinolonas en el tratamiento de la fiebre tifoidea y se discute la fisiopatología del paciente

La enfermería en la atención primaria / Nursing in primary attention

Se expone brevemente la evolución de la atención comunitaria en nuestro país, donde se resalta el trabajo del enfermero y los cambios cuantitativos y cualitativos que se han producido en el personal de Enfermería. Se señalan las cifras de enfermeros dedicados a los servicios de atención primaria y en especial en el plan del médico de la familia. Se esbosa la formación de recursos humanos, la matrícula actual de estudiantes de Enfermería en cursos básicos, y los cambios significativos en el nuevo plan de estudio del curso posbásico en la modalidad de curso dilatado y de la Licenciatura en Enfermería en curso regular de trabajadores por encuentro, con lo cual se dará respuesta al amplio pérfil de trabajo en la atención a grupos etarios y las acciones directas en el medio familiar y social. Se destaca la necesidad de los diferentes modelos de atención primaria que han surgido en estos últimos años, los cuales estan en correspondencia con el desarrollo socioeconómico del país

Planificación de los recursos humanos en enfermería / Planning of human resources in nursing

Se muestran algunos indicadores de enfermería basados en el cálculo de camas y de habitantes según los documentos que norman la plantilla del Sistema Nacional de Salud. En los antecedentes se refleja el desarrollo en la formación del personal de enfermería en correspondencia con los programas de salud y las unidades, desde la creación del Sistema Nacional de Salud al triunfo de la Revolución. Se plantea el procedimiento actual para el cálculo de las plantillas por categorías ocupacionales y cargos. Se señala cómo están agrupadas las normativas. Se dan ejemplos para determinar el número de enfermeros necesarios por especialidades y áreas de trabajo mediante la aplicación de los documentos que norman esto así como su distribución por turno de trabajo. Se expone el sistema de normas para la formación de enfermeros