Characterization of the C6D7-RBD Human Monoclonal Antibody Specific to the SARS-CoV-2 S Protein Receptor-Binding Domain.

The new coronavirus infection COVID-19 is an acute viral disease that affects primarily the upper respiratory tract. The etiological agent of COVID-19 is the SARS-CoV-2 RNA virus (Coronaviridae family, Betacoronavirus genus, Sarbecovirus subgenus). We have developed a high-affinity human monoclonal antibody, called C6D7-RBD, which is specific to the S protein receptor-binding domain (RBD) from the SARS-CoV-2 Wuhan-Hu-1 strain and exhibits virus-neutralizing activity in a test with recombinant antigens: angiotensin-converting enzyme 2 (ACE2) and RBD.

Production and Characterization of Rat Monoclonal Antibodies against the PAL Antigen of Legionella spp.

The purpose of this work was to obtain genus-specific monoclonal antibodies against the Legionella spp. recombinant PAL protein, which will subsequently allow to use them as a basis for the development of new express tests for pathogenic legionella detection. A short three-week immunization protocol for Wistar rats was used to generate rat-mouse heterohybridomas producing antibodies against PAL. Mouse myeloma cell line Sp2/0-Ag14 served as the fusion partner. Hybridization was performed using two methods: PEG-mediated fusion and electrofusion. Subsequent screening was performed by indirect solid-phase ELISA against the target protein rPAL. Specificity analysis was performed by dot-blot using a panel of lysates obtained from 39 pure cultures of different strains, which included closely related and heterologous microorganisms among others. No difference in the efficiency of stable hybridoma clones production by the two indicated cell-fusion methods was detected. Twelve clones producing specific rat monoclonal antibodies were obtained based on the screening results. The obtained rat monoclonal antibodies are highly specific towards the PAL protein of L. pneumophila of different serological groups and other pathogenic legionella and are good candidates to be used as the components of diagnostic test systems for the detection of pathogenic representatives of the Legionella genus.

Mechanism of Action of Monoclonal Antibodies That Block the Activity of the Lethal Toxin of Bacillus Anthracis.

Neutralization of the lethal toxin of Bacillus anthracis is an important topic of both fundamental medicine and practical health care, regarding the fight against highly dangerous infections. We have generated a neutralizing monoclonal antibody 1E10 against the lethal toxin of Bacillus anthracis and described the stages of receptor interaction between the protective antigen (PA) and the surface of eukaryotic cells, the formation of PA oligomers, assembly of the lethal toxin (LT), and its translocation by endocytosis into the eukaryotic cell, followed by the formation of a true pore and the release of LT into the cell cytosol. The antibody was shown to act selectively at the stage of interaction between Bacillus anthracis and the eukaryotic cell, and the mechanism of toxin-neutralizing activity of the 1E10 antibody was revealed. The interaction between the 1E10 monoclonal antibody and PA was found to lead to inhibition of the enzymatic activity of the lethal factor (LF), most likely due to a disruption of true pore formation by PA, which blocks the release of LF into the cytosol.

DNA Barcoding of an Assembly of Montane Andean Butterflies (Satyrinae): Geographical Scale and Identification Performance.

DNA barcoding is a technique used primarily for the documentation and identification of biological diversity based on mitochondrial DNA sequences. Butterflies have received particular attention in DNA barcoding studies, although varied performance may be obtained due to different scales of geographic sampling and speciation processes in various groups. The montane Andean Satyrinae constitutes a challenging study group for taxonomy. The group displays high richness, with more of 550 species, and remarkable morphological similarity among taxa, which renders their identification difficult. In the present study, we evaluated the effectiveness of DNA barcodes in the identification of montane Andean satyrines and the effect of increased geographical scale of sampling on identification performance. Mitochondrial sequences were obtained from 104 specimens of 39 species and 16 genera, collected in a forest remnant in the northwest Andes. DNA barcoding has proved to be a useful tool for the identification of the specimens, with a well-defined gap and producing clusters with unambiguous identifications for all the morphospecies in the study area. The expansion of the geographical scale with published data increased genetic distances within species and reduced those among species, but did not generally reduce the success of specimen identification. Only in Forsterinaria rustica (Butler, 1868), a taxon with high intraspecific variation, the barcode gap was lost and low support for monophyly was obtained. Likewise, expanded sampling resulted in a substantial increase in the intraspecific distance in Morpho sulkowskyi (Kollar, 1850); Panyapedaliodes drymaea (Hewitson, 1858); Lymanopoda obsoleta (Westwood, 1851); and Lymanopoda labda Hewitson, 1861; but for these species, the barcode gap was maintained. These divergent lineages are nonetheless worth a detailed study of external and genitalic morphology variation, as well as ecological features, in order to determine the potential existence of cryptic species. Even including these cases, DNA barcoding performance in specimen identification was 100% successful based on monophyly, an unexpected result in such a taxonomically complicated group.

Graphita gen. nov., a New Genus for Neonympha griphe C. Felder & R. Felder, 1867 (Lepidoptera, Nymphalidae, Satyrinae).

A new genus is described for Neonympha griphe C. Felder & R. Felder, 1867, to contribute toward a revision of the polyphyletic genus Euptychoides Forster, 1964. Based on DNA sequence data, Graphita Nakahara, Marín & Barbosa, gen. nov. is strongly supported as a member of a clade of predominantly southeastern Brazilian taxa, in which it is weakly supported as sister to a well-supported clade containing Pharneuptychia Forster, 1964, Moneuptychia Forster, 1964 and the E. castrensis (Schaus, 1902) species complex. The data show that Graphita griphe comb. nov. is not related to other Euptychoides and not very closely related to any other sampled euptychiines, and thus support the description of this new genus. In addition, we provide morphological illustrations and a distribution map for this taxon based on museum specimens.

[Perceptions and experiences of access to health services and their utilization among the immigrant population]. / Percepción y experiencias en el acceso y el uso de los servicios sanitarios en población inmigrante.

OBJECTIVE: To identify and describe the needs and problems of the immigrant population related to access and utilization of health services. METHOD: A descriptive, qualitative, phenomenological study was conducted using focus groups. The study area was the county of Campo de Gibraltar (Spain), which represents the gateway to Europe for immigration from Africa. The final sample size (51 immigrants from 11 countries) was determined by theoretical saturation. A narrative analysis was conducted with QSR NVivo9 software. RESULTS: Immigrants' discourse showed four categories of analysis: response to a health problem, system access, knowledge of social and health resources, and health literacy needs. Responses to health problems and the route of access to the health care system differed according to some sociodemographic characteristics (nationality/culture of origin, length of residence, and economic status). In general, immigrants primarily used emergency services, hampering health promotion and prevention. The health literacy needs identified concerned language proficiency and the functioning of the health system. CONCLUSIONS: There is a need to promote interventions to enhance health literacy among immigrants. These interventions should take into account diversity and length of residence, and should be based on an action-participation methodology.

Morphological and molecular marker contributions to disentangling the cryptic Hermeuptychia hermes species complex (Nymphalidae: Satyrinae: Euptychiina).

The genus Hermeuptychia is common and widespread through the Americas, from Argentina to the southern United States of America. All eight recognized species within Hermeuptychia are small and brown, with very similar interspecific external morphologies and intraspecifically variable ocelli patterns that render taxonomic identification based on morphology difficult. In our study, we surveyed variability within Hermeuptychia, and evaluated species boundaries based on molecular data (sequences of the 'barcode' mitochondrial DNA COI gene) and morphology (mainly male genitalia), using a phylogenetic approach. We found eight DNA-based and 12 morphological groups in our sampling. Species names were assigned based mainly on comparisons with male genitalia morphology descriptions corresponding to name-bearing type specimens. Morphological and DNA variability were highly congruent, with the exception of group H, the Hermeuptychia cucullina complex. Also, the barcode region showed a clear threshold for intra- and interspecific mean distances around 2%. Based on these results, we circumscribe the species boundaries in the genus Hermeuptychia and discuss conflicts between mitochondrial genes and classic morphological approaches for identifying and delimiting species. Our study revealed cryptic diversity within an ubiquitous genus of Neotropical butterflies.

Long-term evaluation of phrenic nerve pacing for respiratory failure due to high cervical spinal cord injury.

OBJECTIVES: Phrenic nerve pacing is a method of respiratory support that can replace mechanical ventilation in high-level cervical spinal cord injury patients with diaphragmatic paralysis. Our objective was to evaluate survival and long-term quality of life in patients with external respiratory support by PNP vs volumetric respirator in patients with severe respiratory insufficiency due to a high-level spinal cord injury. DESIGN: This is a retrospective review study of a prospectively collected database for evaluate the survival and a questionnaire for quality of life has been collected face-to-face or by telephone at present. PATIENTS: Cervical SCI patients with permanent respiratory support (PNP or MV). METHODS: Long-term evaluation of a cohort of PNP-supported patients. We performed a comparison between these patients and volumetric respirator-supported patients. For survival analysis, we used the Kaplan-Meier method and Cox proportional hazards model. The health-related quality of life was assessed with SF-36 questionnaire, a general HRQL evaluation. RESULTS: One hundred twenty six patients on permanent respiratory support were evaluated during the study period. Of these, 38 were on PNP and 88 were mechanically ventilated. Paced patients were younger and had a longer survival, but in a multivariate analysis adjusted for age using a multiple logistic correlation we found that length of survival was greater for PNP patients. In terms of HRQL, the PNP-supported patients showed better results in terms of social functioning. CONCLUSIONS: PNP is a stable and effective method of long-term respiratory support in this type of patients (SCI patients dependent on external respiratory support). In these patients it improves the length of survival and some social issues by quality of life when compared with patients under MV.

Euptychia boulleti (Le Cerf) n. comb. (Lepidoptera: Nymphalidae: Satyrinae), a rare and endangered butterfly from Southeastern Brazil.

This paper discusses the systematic position of the rare and endangered satyrine butterfly Caenoptychia boulleti Le Cerf, the only included species in Caenoptychia (type species), based on adult morphology and molecular data. The results showed that Caenoptychia Le Cerf belongs to the Euptychia Hübner clade, and the genus is synonymized with Euptychia, new synonymy. Euptychia boulleti (Le Cerf) is a new combination. The male genitalia of E. boulleti showed at least one important synapomorphy with the other species of Euptychia, which is the presence of a posterior projection of the tegumen above the uncus. Molecular data reinforces the position of Caenoptychia within the genus Euptychia.

[Factors explaining the application of the law of patient autonomy by surgical staff]. / Factores explicativos de la aplicación de la Ley de autonomía del paciente por personal médico y de enfermería del área quirúrgica.

OBJECTIVE: To examine which contextual factors explain the implementation of the Patient Autonomy Law (LAP) by professionals involved in the surgical process. METHODS: A descriptive cross sectional study survey. It was conducted at the Hospital Virgen Macarena in Seville with the surgical staff. Dependent variable: the application of LAP. INDEPENDENT VARIABLES: the perception of socio-structural organizational characteristics according to Kanter's model (organizational culture, sociopolitical support, access to information, and access to resources), the perception of interprofessional collaboration, and knowledge of the LAP. We performed exploratory analysis, bivariate with parametric and nonparametric tests depending on the nature of the distributions; simple regression to determine the weight of each explanatory variable on the application of the LAP and, multiples introducing in the models those variables that had obtained significant regression coefficients. The analysis performed with a confidence level of 95%. PARTICIPANTS: 170 professionals (response rate: 60.28%). Behaviour according to the LAP is present in a medium-high level among physicians and nurses. Significant differences were observed in the application of the LAP according to the different sociolaboral variables and considering medical staff, the working environment and the specialty. Multiple regression analyses revealed the relationship between the application of the LAP and interprofessional collaboration, in physicians. CONCLUSIONS: Kanter's model and the scale of interprofessional collaboration, may contribute to the explanation of the influence of the hospital context in the implementation of the LAP and the results may guide us when designing strategies to implement the law. It would be necessary to overcome the traditional structures and create decentralized and flexible conditions that encourage teamwork and shared decision making.

From the phylogeny of the Satyrinae butterflies to the systematics of Euptychiina (lepidoptera: nymphalidae): history, progress and prospects.

We review the various proposals of evolutionary and classification schemes for Satyrinae and particularly Euptychiina butterflies, assessing progress and prospects of research for the group. Among the highlights is the proposal to include Morphini, Brassolini and Amathusiini as part of Satyrinae. Although it is clear that this hypothesis requires further investigation, phylogenetic studies recently conducted recover this clade as part of Satyrinae with high support. The phylogenetic analyses for Euptychiina carried out to date recover the monophyly of the group and have identified a variety of genera as non-monophyletic. Further work is necessary to resolve the position of the subtribe and the evolutionary relationships of several genera.

Insuficiencia del ligamento cruzado anterior. Valor diagnóstico de la exploración clinica y RM / Anterior cruciate ligament failure: Diagnostic value of the clinical examination and magnetic resonance imaging

Objetivo. El diagnóstico clínico de la insuficiencia del LCA está sometido a criterios subjetivos y su sensibilidad es baja. La RM se ha convertido en el estándar en la evaluación no invasiva de las lesiones de rodilla. Es importante determinar el valor diagnóstico de la exploración clínica (EC) y resonancia magnética (RM) frente al patrón artroscopia en lesiones del LCA y conocer y precisar sus parámetros exploratorios objetivos. Material y método. Se han revisado 101 H. Clínicas con las condiciones: EC-radiológica sospechosa de patología LCA, exploración protocolizada con test subjetivos/objetivos, RMN, EC objetiva bajo anestesia y artroscopia diagnóstica/terapéutica. Se seleccionaron datos epidemiológicos, tiempos desde la lesión del LCA hasta la cirugía, intervenciones previas sobre rodilla afecta, datos subjetivos, datos objetivos incluyendo artrometría, número de centros que realizaron la RM, número de RM por centro y observadores, y artroscopia. Material y método. Se ha realizado estudio estadístico comparando variables cuantitativas y cualitativas, precisión, fiabilidad y consistencia entre medidas. Resultados. Respectivamente el 94%, el 100% y el 83% fueron diagnosticados mediante EC-sin-anestesia, con-anestesia y RM, con una sensibilidad del 94,06%, 100%, y 83,17%. Discusión. La EC, con/sin-anestesia, diagnóstica más veces la insuficiencia del LCA que la RM. La RM diagnostica un número total mayor de lesiones concomitantes y detecta más lesiones del menisco externo. El test de Lachman y la artrometría son los métodos diagnósticos más significativos en la insuficiencia del LCA (AU)

Objective. The clinical diagnosis of anterior cruciate ligament (ACL) failure uses subjective criteria and its sensitivity is low. Magnetic resonance imaging (MRI) has become the standard in the non-invasive evaluation of knee injuries. It is important to determine the diagnosis by clinical examination (CE) and MR versus the arthroscopy pattern in the ACL injuries, and to know and specify their objective exploratory parameters. Materials and Methods. A total of 101 medical records were reviewed: CE-radiological suspicion of ACL failure, examination using a subjective/objective test protocol, MR, objective CE under anaesthesia and diagnostic/therapeutic arthroscopy. We selected epidemiological data, times from the ACL to surgery, previous operation on the affected knee, subjective data, objective data including arthrometry, number of centres that performed MR, the number of MR per centre, observations and arthroscopy. Results. A statistical study was performed comparing quantitative and qualitative variables, precision, reliability and consistency between actions. A total of 94%, 100% and 83% were diagnosed using EC without anaesthesia, with anaesthesia and MR, with a sensitivity of 94.06%, 100% and 83.17%, respectively. Discussion. The CE with/without-anaesthesia, diagnosed ACL failure more often than MR. MR diagnoses the total number of concomitant injuries and detects more external meniscus injuries. The Lachman test and arthrometry are the most significant diagnostic methods in ACL failure (AU)