Green bottle flies (Calliphoridae, Luciliinae) of Ecuador: geographic distribution, checklist and DNA barcodes

Green bottle flies (Diptera, Calliphoridae, Luciliinae) comprise a diverse and cosmopolitan taxon, known from at least 1,500 species. They have become crucial elements in forensic investigations, as they spend part of their life cycle in decaying remains. Here, we review the distribution of eleven Luciliinae species in Ecuador: the monotypic Blepharicnema and ten Lucilia species. We identified specimens using morphological characters. Additionally, we DNA barcoded 43 specimens from three species using 658bp segments of the standard Cytochrome Oxidase I (COI) mitochondrial gen. Molecular and morphological identifications presented high correspondence, suggesting COI barcodes are an efficient tool for the identification of these three green bottle flies species. Geographical records are biased towards the northern Andean region, particularly near to large urban settlements. We remark the value to applied forensic research of continuous sampling of necrophagous flies under a variety of habitats and crime conditions.Fil: Moreno Coellar, Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina. Pontificia Universidad Católica del Ecuador; EcuadorFil: Amat, Eduardo. Tecnológico de Antioquia; ColombiaFil: Meneses, Pablo E.. Universidad Regional Amazónica Ikiam; EcuadorFil: Donoso, David A.. Escuela Politécnica Nacional; EcuadorFil: Barragán, Álvaro R.. Pontificia Universidad Católica del Ecuador; Ecuado

Diversity among clinical isolates of penicillin-resistant Streptococcus mitis: indication for a PBP1-dependent way to reach high levels of penicillin resistance

A total of 12 non-epidemiologically related clinical isolates of Streptococcus mitis that showed different levels of resistance to penicillin were studied. Membrane-protein profiles and penicillin-binding protein (PBP) patterns showed a great polymorphism; and patterns of 4—7 PBPs, withsizes that ranged from ~101 kDa to ~40 kDa, were detected in eachstrai n. No association could be found between PBP pattern and resistance level to penicillin among these isolates. Arbitrarily primed PCR confirmed the genetic diversity among this group of streptococci. One of the isolates of intermediate level of resistance to penicillin, which showed a PBP pattern similar to that of the high-resistance strains, was used as a laboratory model to analyse the mechanism underlying high-resistance acquisition by these strains. A 14-fold increase in penicillin resistance was obtained after a single selection step, which resulted in a decrease in penicillin affinity for PBP1. The size of this PBP (92 kDa) and the differences in PBP profiles of the penicillin-resistant clinical isolates suggest the existence in S. mitis of PBP-mediated mechanisms to acquire high-level resistance to penicillin, among which alterations in PBP1 seem to play a main role, in contrast to the PBP2X mediated mechanism described for other streptococci

Dietary Intervention Modulates the Expression of Splicing Machinery in Cardiovascular Patients at High Risk of Type 2 Diabetes Development: From the CORDIOPREV Study

Type-2 diabetes mellitus (T2DM) has become a major health problem worldwide. T2DM risk can be reduced with healthy dietary interventions, but the precise molecular underpinnings behind this association are still incompletely understood. We recently discovered that the expression profile of the splicing machinery is associated with the risk of T2DM development. Thus, the aim of this work was to evaluate the influence of 3-year dietary intervention in the expression pattern of the splicing machinery components in peripheral blood mononuclear cells (PBMCs) from patients within the CORDIOPREV study. Expression of splicing machinery components was determined in PBMCs, at baseline and after 3 years of follow-up, from all patients who developed T2DM (Incident-T2DM, n = 107) and 108 randomly selected non-T2DM subjects, who were randomly enrolled in two healthy dietary patterns (Mediterranean or low-fat diets). Dietary intervention modulated the expression of key splicing machinery components (i.e., up-regulation of SPFQ/RMB45/RNU6, etc., down-regulation of RNU2/SRSF6) after three years, independently of the type of healthy diet. Some of these changes (SPFQ/RMB45/SRSF6) were associated with key clinical features and were differentially induced in Incident-T2DM patients and non-T2DM subjects. This study reveals that splicing machinery can be modulated by long-term dietary intervention, and could become a valuable tool to screen the progression of T2DM

Los factores ambientales, luz y temperatura, modifican la formación de la biopelícula en Pseudomonas syringae pv. syringae.

Comunicación a congreso en formato pósterLas biopelículas bacterianas están formadas por agregados celulares embebidos en una matriz extracelular de producción propia, formada principalmente por exopolisacáridos (EPS), proteínas y ADN extracelular. Existen datos recientes que sugieren que entre el 40-80% de las bacterias se encuentran formando biopelículas en la naturaleza, y que la formación de las biopelículas se ve afectada por factores ambientales. Pseudomonas syringae pv. syringae (Pss) es una bacteria fitopatógena causante de la necrosis apical del mango (NAM), enfermedad limitante de la producción de este cultivo en la región mediterránea. En Pss se ha descrito una conexión entre factores ambientales y la incidencia y severidad de la NAM, jugando los EPS celulosa y uno tipo-Psl, un papel relevante en la transición entre un estilo de vida epífito o patogénico. En este trabajo, se ha analizado el papel de algunos factores ambientales relevantes como son la luz y la temperatura en la formación de la biopelícula in vitro en una colección de cepas de Pss aisladas de mango. Los resultados obtenidos sugieren que la temperatura y particularmente la luz pueden influir en la formación de las biopelículas a través de la biosíntesis de los EPS, principalmente celulosa. Además, también se ha observado como niveles más bajos de producción de celulosa podrían estar asociados con una mayor virulencia. Estos resultados indican que los EPS, y principalmente celulosa juegan un papel clave en la ecología de Pss sobre la planta de mango a través de la formación de la biopelícula.Este trabajo ha sido financiado por Proyectos de Excelencia de la Junta de Andalucía (P12- AGR-1473), cofinanciado con ayudas a proyectos I+D+i en el marco del Programa Operativo FEDER Andalucía (UMA20-FEDERJA-060), y la Universidad de Málaga. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Nephrin mutations cause childhood- and adult-onset focal segmental glomerulosclerosis

Mutations in the NPHS1 gene cause congenital nephrotic syndrome of the Finnish type presenting before the first 3 months of life. Recently, NPHS1 mutations have also been identified in childhood-onset steroid-resistant nephrotic syndrome and milder courses of disease, but their role in adults with focal segmental glomerulosclerosis remains unknown. Here we developed an in silico scoring matrix to evaluate the pathogenicity of amino-acid substitutions using the biophysical and biochemical difference between wild-type and mutant amino acid, the evolutionary conservation of the amino-acid residue in orthologs, and defined domains, with the addition of contextual information. Mutation analysis was performed in 97 patients from 89 unrelated families, of which 52 presented with steroid-resistant nephrotic syndrome after 18 years of age. Compound heterozygous or homozygous NPHS1 mutations were identified in five familial and seven sporadic cases, including one patient 27 years old at onset of the disease. Substitutions were classified as ‘severe’ or ‘mild’ using this in silico approach. Our results suggest an earlier onset of the disease in patients with two ‘severe’ mutations compared to patients with at least one ‘mild’ mutation. The finding of mutations in a patient with adult-onset focal segmental glomerulosclerosis indicates that NPHS1 analysis could be considered in patients with later onset of the disease

Polarimetric imaging for the detection of synthetic models of SARS-CoV-2: A proof of concept

Objective: To conduct a proof-of-concept study of the detection of two synthetic models of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using polarimetric imaging. Approach: Two SARS-CoV-2 models were prepared as engineered lentiviruses pseudotyped with the G protein of the vesicular stomatitis virus, and with the characteristic Spike protein of SARS-CoV-2. Samples were prepared in two biofluids (saline solution and artificial saliva), in four concentrations, and deposited as 5-µL droplets on a supporting plate. The angles of maximal degree of linear polarization (DLP) of light diffusely scattered from dry residues were determined using Mueller polarimetry from87 samples at 405 nm and 514 nm. A polarimetric camera was used for imaging several samples under 380–420 nm illumination at angles similar to those of maximal DLP. Per-pixel image analysis included quantification and combination of polarization feature descriptors in 475 samples. Main results: The angles (from sample surface) of maximal DLP were 3° for 405 nm and 6° for 514 nm. Similar viral particles that differed only in the characteristic spike protein of the SARS-CoV-2, their corresponding negative controls, fluids, and the sample holder were discerned at 10-degree and 15-degree configurations. Significance: Polarimetric imaging in the visible spectrum may help improve fast, non-contact detection and identification of viral particles, and/or other microbes such as tuberculosis, in multiple dry fluid samples simultaneously, particularly when combined with other imaging modalities. Further analysis including realistic concentrations of real SARS-CoV-2 viral particles in relevant human fluids is required. Polarimetric imaging under visible light may contribute to a fast, cost-effective screening of SARS-CoV-2 and other pathogens when combined with other imaging modalities.12 página

Hyperspectral image processing for the identification and quantification of lentiviral particles in fluid samples

Optical spectroscopic techniques have been commonly used to detect the presence of biofilm-forming pathogens (bacteria and fungi) in the agro-food industry. Recently, near-infrared (NIR) spectroscopy revealed that it is also possible to detect the presence of viruses in animal and vegetal tissues. Here we report a platform based on visible and NIR (VNIR) hyperspectral imaging for non-contact, reagent free detection and quantification of laboratory-engineered viral particles in fluid samples (liquid droplets and dry residue) using both partial least square-discriminant analysis and artificial feed-forward neural networks. The detection was successfully achieved in preparations of phosphate buffered solution and artificial saliva, with an equivalent pixel volume of 4 nL and lowest concentration of 800 TU.mu L-1. This method constitutes an innovative approach that could be potentially used at point of care for rapid mass screening of viral infectious diseases and monitoring of the SARS-CoV- 2 pandemic.This research was funded by grants number COV20-00080 and COV20-00173 of the 2020 Emergency Call for Research Projects about the SARS-CoV-2 virus and the COVID-19 disease of the Institute of Health 'Carlos III', Spanish Ministry of Science and Innovation, and by grant number EQC2019-006240-P of the 2019 Call for Acquisition of Scientific Equipment, FEDER Program, Spanish Ministry of Science and Innovation. This work has been supported by the European Commission through the JRC HUMAINT project. ABR was supported by grant number RTI2018-094465-J funded by the Spanish National Agency of Research. The authors would like to gratefully acknowledge the assistance of the members of the EOD-CBRN Group of the Spanish National Police, whose identities cannot be disclosed, and who are represented here by JMNG. Authors thank continuous support from their institutions