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MolPredictX is a free-access web tool in which it is possible to analyze the prediction of biological activity of chemical molecules. MolPredictX has been available online to the general public for just over a year and has now gone through its first update. We also developed its version for android, being the first free app capable of predicting biological activities. MolPredictX is available for free at https://www.molpredictX.ufpb.br/ , and its mobile application version can be obtained from Google Play.

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Aprendizaje Automático , Aplicaciones Móviles , Programas Informáticos , Internet , Biología Computacional/métodos , Humanos

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This study examines the influence of body mass index (BMI) on the relationship between quantitative sensory testing measures and clinical characteristics in fibromyalgia syndrome (FMS). Utilizing BMI as a categorical covariate (≥25 or ≥30 kg/m²) in associations between quantitative sensory testing metrics (pain-60, conditioned pain modulation, and temporal summation of pain [TSP]) and FMS clinical features, we explored BMI's role as both a confounder (change-in-estimate criterion-change equal or higher than 10%) and effect modifier (interaction term). Significant interactions revealed overweight/obese BMI as a modifier in the relationship between conditioned pain modification and both depression and symptom impact, with a homeostatic relationship between better clinical profile and pain inhibitory response observed solely in the normal-weight group. Similar results were found for pain-60 and depression. Additionally, BMI ≥30 kg/m² modified TSP's effect on pain, demonstrating lower pain with increased TSP, exclusively in the nonobese group. This study highlights the significant role of BMI in moderating the relationships of important pain inhibitory control processes and pain intensity, depression, and the overall impact of FMS symptoms. Our results suggest that high BMI states disrupt the homeostatic effects of pain inhibition, reducing its salutogenic response in FMS participants. We discuss the mechanistic and therapeutic implications of targeting BMI in FMS clinical trials and the potential impact of this important relationship. PERSPECTIVE: This investigation highlights the disruptive influence of high BMI on pain inhibitory control in fibromyalgia, unbalancing clinical symptoms such as pain and depression. It underscores the necessity of integrating BMI considerations into therapeutic approaches to enhance pain management and patient outcomes.

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Introduction: Photobiomodulation with low-level laser treatment can enhance bone formation by stimulating the cell division of osteoblasts and increasing the amount of protein deposition, thus encouraging the formation of new bone. The aim of this study was to evaluate the effects of photobiomodulation with a low-level laser on proliferation and gene expression related to calcium signaling in human osteoblasts. Methods: Osteoblastic cell lines of the hFOB1.19 lineage, human osteoblasts, were grown and assigned into two groups, control (C; n=78 cultured wells) and photobiomodulation (L; n=78 cultured wells) with n=6 per day of the experimental period. Cells were cultured (immature at 34 ºC), and after maturation at 37 ºC, group L cells were exposed to laser irradiation with a low-level laser device (gallium and aluminum arsenide), at a wavelength of 808 nm, a power output of 200 mW, and a power density of 200 mW/cm2. The energy delivered to the cells was 37 J/cm2, with a beam area of 0.02 mm2 and an exposure time of 5 seconds. This treatment was applied daily for a period of 13 days. Following this, the number of cells was counted, and RNA was isolated, measured, and then converted into cDNA for further quantification using a comparative Ct method with real-time polymerase chain reaction. The results were then subjected to statistical analysis through a Mann-Whitney test, with a significance level of P<0.05. Results: The cell count in the L group (37.25x10±4±22.02) was statistically higher compared to the control group (22.75x10±4±7.660) with a P value of 0.0259. The values of 2-ΔΔCt for S100A6, plasma membrane calcium ATPase (PMCA), and calmodulin genes indicated hyper-expression on the thirteenth day, while the osteocalcin gene showed hypo-expression. Conclusion: The study suggests that the photobiomodulation mechanism with a low-level laser may regulate gene expression in human osteoblasts in a dose-dependent and cumulative manner.

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Cell replacement therapies using medial ganglionic eminence (MGE)-derived GABAergic precursors reduce seizures by restoring inhibition in animal models of epilepsy. However, how MGE-derived cells affect abnormal neuronal networks and consequently brain oscillations to reduce ictogenesis is still under investigation. We performed quantitative analysis of pre-ictal local field potentials (LFP) of cortical and hippocampal CA1 areas recorded in vivo in the pilocarpine rat model of epilepsy, with or without intrahippocampal MGE-precursor grafts (PILO and PILO+MGE groups, respectively). The PILO+MGE animals had a significant reduction in the number of seizures. The quantitative analysis of pre-ictal LFP showed decreased power of cortical and hippocampal delta, theta and beta oscillations from the 5 min. interictal baseline to the 20 s. pre-ictal period in both groups. However, PILO+MGE animals had higher power of slow and fast oscillations in the cortex and lower power of slow and fast oscillations in the hippocampus compared to the PILO group. Additionally, PILO+MGE animals exhibited decreased cortico-hippocampal synchrony for theta and gamma oscillations at seizure onset and lower hippocampal CA1 synchrony between delta and theta with slow gamma oscillations compared to PILO animals. These findings suggest that MGE-derived cell integration into the abnormally rewired network may help control ictogenesis.

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Corteza Cerebral , Modelos Animales de Enfermedad , Epilepsia , Hipocampo , Pilocarpina , Animales , Pilocarpina/toxicidad , Hipocampo/fisiopatología , Masculino , Corteza Cerebral/fisiopatología , Epilepsia/inducido químicamente , Epilepsia/fisiopatología , Ratas , Ondas Encefálicas/fisiología , Ratas Wistar , Electroencefalografía , Eminencia Ganglionar

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AIM: This single-arm interventional trial aimed to investigate the efficacy of ultrasonic irrigation as a supplementary disinfection approach after chemomechanical procedures using molecular techniques based on ribosomal RNA (rRNA) and rRNA genes (referred to as DNA). METHODOLOGY: Samples were collected from 35 single-rooted teeth with radiographic evidence of apical periodontitis. Samples were taken after gaining root canal access (S1), chemomechanical procedures (CMP, S2), and ultrasonic irrigation (S3). DNA-targeted qPCR using universal primers was used to estimate total bacterial levels, while rRNA-targeted qPCR was used to assess bacterial activity. Ratios between rRNA and DNA levels were calculated to search for active bacteria in the samples (rRNA/ DNA ≥ 1). Wilcoxon matched-pairs signed-rank test was used to compare the differences in DNA levels between samples and DNA and rRNA levels within samples (P <.05). RESULTS: DNA-based methods revealed a significant decrease in bacterial levels from S1 to S2 and S2 to S3 (both P <.05). Notably, 11 out of 35 (31.4%) root canals did not harbor bacterial DNA after CMP, whereas ultrasonic activation increased DNA-negative samples to 17 (48.6%). However, all DNA-positive samples were also positive for rRNA, with significantly higher rRNA than DNA levels (P <.05), indicating bacterial activity at the sampling time. CONCLUSIONS: Ultrasonic irrigation improved the disinfection of root canals after chemomechanical procedures by reducing bacterial levels. However, persisting bacteria remained active in the root canals after CMP and ultrasonic irrigation.

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Dystrophin Dp71 is essential for the development of the nervous system. Its alteration is associated with intellectual disability. Different Dp71 isoforms are generated by alternative splicing; however, their functions have not been fully described. Here, we identified Dp71dΔ71-associated proteins to understand the complex functions. PC12 cells, stably transfected with pTRE2pur-Myc/Dp71dΔ71 or pTRE2pur-Myc empty vector (EV), were analyzed by immunoprecipitation followed with quantitative proteomics with data-independent acquisition and ion mobility separation. We used the Top3 method to quantify absolutely every protein detected. A total of 106 proteins were quantified with Progenesis QI software and the database UP000002494. Seven new proteins associated with Dp71dΔ71 were selected with at least 2-fold quantity between immunoprecipitated proteins of PC12-Myc/Dp71dΔ71 versus PC12-EV cells. These results revealed new proteins that interact with Dp71dΔ71, including ß-Tubulin, S-adenosylmethionine synthase isoform type-2, adapter molecule crk, helicase with zinc finger 2, WD repeat domain 93, cyclin-L2 and myosin-10, which are related to cell migration and/or cell growth. The results lay the foundation for future research on the relationship between these proteins and Dp71 isoforms.

RESUMEN

PREMISE: Understanding the responses of functional traits in tree species to climate variability is essential for predicting the future of tropical montane cloud forest (TMCF) tree species, especially in Andean montane environments where fog pockets act as moisture traps. METHODS: We studied the distribution of Magnolia gentryi, measured its spatial arrangement, identified local hotspots, and evaluated the extent to which climate-related factors are associated with its distribution. We then analyzed the variation in 13 functional traits of M. gentryi and the relationship with climate. RESULTS: Andean TMCF climatic factors constrain M. gentryi spatial distribution with significant patches or gaps that are associated with high precipitation and mean minimum temperature. The functional traits of M. gentryi are limited by the Andean TMCF climatic factors, resulting in reduced within-species variation in traits associated with water deficit. CONCLUSIONS: The association between functional traits and climate oscillation is crucial for understanding the growth conditions of relict-endemic species and is essential for conservation efforts. Forest trait diversity and species composition change because of fluctuations in hydraulic safety-efficiency gradients.

Asunto(s)

Bosques , Magnolia , Clima Tropical , Perú , Magnolia/fisiología , Magnolia/genética , Cambio Climático , Árboles/fisiología

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BACKGROUND: Emerging evidence suggests that individuals with chronic non-specific neck pain may experience altered sensory processing, potentially contributing to the modest response to therapeutic exercise treatments. OBJECTIVE: This systematic review aims to explore the effect of therapeutic exercise on pain processing among patients with chronic non-specific neck pain. METHODS: A systematic search was conducted in multiple databases (PubMed, EMBASE, CINAHL, PEDro, SportDiscus, and Cochrane CENTRAL) from inception to June 2023. Inclusion criteria included randomized controlled trials (RCT) comparing therapeutic exercise to non-exercise treatments or no treatment. The screening and data extraction was conducted by two reviewers. The methodological quality was evaluated using the PEDro scale and the certainty of evidence using GRADE. The primary outcomes assessed were pressure pain threshold (PPT), temporal summation, and conditioned pain modulation. RESULTS: Thirteen trials included a total of 948 participants, with 586 in the exercise therapy group and 362 in the non-exercise group. The therapeutic exercise was not superior to non-exercise treatments for both local and PPT in the immediate (MD = 0.13, 95%CI = -0.18 to 0.43), and short-term follow-up (MD = 0.17, 95%CI = -0.27 to 0.61). In the medium term, therapeutic exercise demonstrated a small effect size in increasing local PPT (Kg/cm2) (MD = 0.64, 95%CI = 0.08 to 1.19) compared to non-exercise interventions. The certainty of evidence for these outcomes was very low. CONCLUSIONS: There is very low certainty of evidence that therapeutic exercise is not superior than non-exercise treatment on pain processing in patients with chronic non-specific neck pain.

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In the search for natural and non-toxic products alternatives to synthetic pesticides, the fumigant and repellent activities of 35 essential oils are predicted in the human head louse (Pediculus humanus capitis) through the Quantitative Structure-Activity Relationships (QSAR) theory. The number of constituents of essential oils with weight percentage composition greater than 1% varies from 1 to 15, encompassing up to 213 structurally diverse compounds in the entire dataset. The 27,976 structural descriptors used to characterizing these complex mixtures are calculated as linear combinations of non-conformational descriptors for the components. This approach is considered simple enough to evaluate the effects that changes in the composition of each component could have on the studied bioactivities. The best linear regression models found, obtained through the Replacement Method variable subset selection method, are applied to predict 13 essential oils from a previous study with unknown property data. The results show that the simple methodology applied here could be useful for predicting properties of interest in complex mixtures such as essential oils.

Asunto(s)

Insecticidas , Aceites Volátiles , Pediculus , Relación Estructura-Actividad Cuantitativa , Aceites Volátiles/química , Aceites Volátiles/farmacología , Pediculus/efectos de los fármacos , Pediculus/química , Animales , Insecticidas/química , Insecticidas/farmacología , Modelos Lineales , Humanos

RESUMEN

Soil contamination by heavy metals (HM) from pesticides poses a serious environmental threat, affecting sustainability and agricultural productivity. Soil enzymes are essential for biochemical reactions such as organic matter decomposition and nutrient cycling and are vital for maintaining soil health. However, the effects of HM on soil enzyme activity are not yet well understood. This study examined the impact of HM contamination on enzymatic stoichiometry in regions with intensive pesticide use. We selected flower cultivation areas with 5 years (CA1) and 10 years (CA2) of pesticide exposure and a native forest area (NFA) as a reference during the dry and rainy seasons. We measured Cd, Cu, Mn, Pb, and Zn levels and employed ecological risk indices to assess contamination levels. We also analyzed enzyme activities (arylsulfatase, ß-glucosidase, acid phosphatase, urease) and enzymatic stoichiometry. CA2 exhibited the highest concentrations of Cd, Cu, and Mn in both periods, while Zn was highest in both CA1 and CA2. CA2 had higher values for all indices, indicating significant contamination. Compared with NFA, arylsulfatase activity was lower in cultivated areas during both periods, suggesting decreased soil quality. We found negative correlations between Cu, Mn, Zn, and arylsulfatase, as well as a reduction in urease with Cd; these elements also increased microbial C limitation. Our findings show that continuous pesticide input increases HM levels and that enzyme activity and stoichiometry are effective bioindicator of soil contamination. This study underscores the urgent need for guidelines to protect soils from prolonged HM buildup.

Asunto(s)

Agricultura , Monitoreo del Ambiente , Metales Pesados , Plaguicidas , Contaminantes del Suelo , Suelo , Contaminantes del Suelo/análisis , Plaguicidas/análisis , Metales Pesados/análisis , Suelo/química

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Purslane (Portulaca oleracea) and spinach (Spinacea oleracea) are species with elevated levels of oxalic acid, an antinutrient that interferes in the bioaccessibility of minerals such as calcium and iron. Evaluating methods to determine oxalic acid content with reduced matrix interference, such as employing Flame Atomic Absorption Spectrometry (FAAS), can enhance the specificity of determinations. The different matrices of purslane (whole plant, leaves, and juice) and spinach (whole plant) were tested using three extraction methods (M1, M2, and M3). The oxalic acid content was evaluated by UV-vis spectrophotometry and FAAS (Flame Atomic Absorption Spectrometry). The absence of the precipitation step in M1 resulted in high levels of oxalic acid in the investigated matrices. The quantification of oxalic acid by FAAS for M2 (6M HCl for 1 hour at 100°C) and M3 (0.25N HCl for 15 minutes at 100°C) in the samples of purslane leaves and spinach whole plants yielded statistically similar results. However, the analysis by UV-vis spectrophotometry for M2 and M3 showed significant discrepancies in all evaluated samples, suggesting interference from colored compounds in the food matrix.•Comparison of methods of extraction•Comparison of UV-vis spectrophotometer and FAAS in the quantification of oxalic acid•Analysis of antinutrients in plant matrices.

RESUMEN

Introduction: Human trophoblastic cell lines, such as BeWo, are commonly used in 2D models to study placental Trypanosoma cruzi infections. However, these models do not accurately represent natural infections. Three-dimensional (3D) microtissue cultures offer a more physiologically relevant in vitro model, mimicking tissue microarchitecture and providing an environment closer to natural infections. These 3D cultures exhibit functions such as cell proliferation, differentiation, morphogenesis, and gene expression that resemble in vivo conditions. Methods: We developed a 3D culture model using the human trophoblastic cell line BeWo and nonadherent agarose molds from the MicroTissues® 3D Petri Dish® system. Both small (12-256) and large (12-81) models were tested with varying initial cell numbers. We measured the diameter of the 3D cultures and evaluated cell viability using Trypan Blue dye. Trophoblast functionality was assessed by measuring ß-hCG production via ELISA. Cell fusion was evaluated using confocal microscopy, with Phalloidin or ZO-1 marking cell edges and DAPI staining nuclei. T. cruzi infection was assessed by microscopy and quantitative PCR, targeting the EF1-α gene for T. cruzi and GAPDH for BeWo cells, using three parasite strains: VD (isolated from a congenital Chagas disease infant and classified as Tc VI), and K98 and Pan4 (unrelated to congenital infection and classified as Tc I). Results: Seeding 1000 BeWo cells per microwell in the large model resulted in comparable cellular viability to 2D cultures, with a theoretical diameter of 408.68 ± 12.65 µm observed at 5 days. Functionality, assessed through ß-hCG production, exceeded levels in 2D cultures at both 3 and 5 days. T. cruzi infection was confirmed by qPCR and microscopy, showing parasite presence inside the cells for all three tested strains. The distribution and progression of the infection varied with each strain. Discussion: This innovative 3D model offers a simple yet effective approach for generating viable and functional cultures susceptible to T. cruzi infection, presenting significant potential for studying the placental microenvironment.

Asunto(s)

Enfermedad de Chagas , Placenta , Trofoblastos , Trypanosoma cruzi , Humanos , Trofoblastos/parasitología , Trypanosoma cruzi/genética , Trypanosoma cruzi/crecimiento & desarrollo , Trypanosoma cruzi/fisiología , Femenino , Embarazo , Placenta/parasitología , Enfermedad de Chagas/parasitología , Línea Celular , Técnicas de Cultivo de Célula/métodos , Supervivencia Celular , Técnicas de Cultivo Tridimensional de Células/métodos

RESUMEN

In this study, nine commercial ice creams and four prototypes were assessed. For the sensory analysis, quantitative descriptive analysis (QDA), and temporal dominance of sensations (TDS) methodology were used. According to the QDA results, full-composition ice cream showed significantly higher differences (p < .05) in terms such as vanilla and sweet flavors, brightness, creamy texture and appearance, and viscosity, exhibiting longer melting times. Functional ice creams showed significantly higher differences (p < .05) in terms like viscosity, creamy texture, and appearance. On the other hand, agave fructans in prototype ice creams were found to be able to reduce fat, but not fat and sugar simultaneously, showing a significant decrease (p < .05) in terms such as hardness, crystallized texture, gummy texture, and porosity. Based on the PCA results, the analysis accounted for 75.28% of data variability. Full-composition ice creams and one functional ice cream were related to terms such as viscosity, fatty sensation, creamy texture and appearance, dense, gummy, among others. The rest of the commercial ice creams were related to vanilla and caramel flavor and smell, artificial aftertaste, aerated, porosity, among others. Prototype ice creams were related to hard texture, salty and milk flavor, and crystallized texture. Based on the results of the TDS method, all formulations were found to be significantly dominant (5%) in terms such as vanilla flavor and sweet flavor at the beginning of the test. Formulations containing butyric fat and/or vegetable fat, or agave fructans were significantly dominant (5%) in fatty sensation.

Asunto(s)

Helados , Gusto , Helados/análisis , Viscosidad , Humanos , Aromatizantes , Agave/química , Fructanos/química , Vanilla/química

RESUMEN

This work aimed to describe and quantify the tissue components of the digestive tube of the neotropical freshwater stingray, Potamotrygon wallacei. For this, conventional histology and stereological methods were used to estimate tissue volume. The volumes of the four fundamental layers and the tissue components in the stomach (cardiac and pyloric) and spiral intestine were also estimated. In the cardiac stomach, the mucosa layer occupies 44.7% of the total volume of the organ wall. The gastric glands are the main components, and these structures alone represent 49.7% of this layer. This large number of gastric glands suggests a high potential for processing food items with a high protein content. The stereological methods were sensitive enough to show a reduction in the volume of the gastric glands from the cardiac region toward the pyloric region. Gastric glands are absent in the pyloric region of the stomach. However, the muscularis becomes thicker towards the pyloric region. The increase in smooth muscle thickness is due to the thickening of the inner muscular layer. This suggests that the role of the pyloric stomach may be related to the mixing of the chyme and assisting its passage to the spiral intestine. In the spiral intestine, data on the volume of the mucosa layer (and epithelial lining) suggest that the spiral valve has a large absorptive area. In several respects, the morphology of the digestive tube of P. wallacei is similar to that of other batoids. However, its slight morphological variations may be related to the habitat specificity of this species.

Asunto(s)

Rajidae , Animales , Rajidae/anatomía & histología , Agua Dulce , Estómago/anatomía & histología , Elasmobranquios/anatomía & histología , Mucosa Gástrica/anatomía & histología , Tracto Gastrointestinal/anatomía & histología , Intestinos/anatomía & histología

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The cocoa tree (Theobroma cacao L.) has seeds that after processing become a valuable agricultural commodity in the world. In Brazil, the state of Pará located in the Eastern Amazon is the main producer, accounting for more than 53% of the national production. Although the region is the largest producer, no studies are found in the literature containing data on the sensory quality of the beans. Thus, the purpose of this study is to establish the sensory profile of Amazonian cocoa from the main cocoa-producing regions of the Eastern Amazon (Lower Tocantins River, Northeast, West, Southeast and Trans-Amazon). The sensory profile was established from the Quantitative Descriptive Analysis (QDA), in addition to using an affective test to verify consumer preference for the chocolates produced. Physical, physicochemical and bioactive compound evaluations were carried out. Beans from different regions had a similar sensory profile; however, it was possible to observe some differences in certain descriptor terms. In the physical evaluation, the samples presented an acceptable commercial standard, and the humidity was within the values established by the legislation. It was found that the post-harvest and/or processing parameters had no influence on the quality attributes of cocoa beans.

RESUMEN

In Brazil, Dengue, Zika and Chikungunya viruses constitute a major threat to the public health system. Simultaneous circulation of these arboviruses occurs in many regions of the world due to the expansion of transmission vectors. The infection by these arboviruses triggers similar symptoms during their acute phase. However, in some cases, severe symptoms may occur, leading to different types of disabilities and even death. In this context, considering the similarity of the symptoms, the problems caused by the infection of these arboviruses, and the increasing risk of coinfection in humans, the differential diagnosis of these infections is essential for clinical management and epidemiological investigation. Thus, this study aimed to identify, through diagnosis via Quantitative Polymerase Chain Reaction with Reverse Transcription, arbovirus coinfection in patients from the Tocantins state (Northern Brazil). A total of 495 samples were analyzed, three from which were determined to be a coinfection of Dengue and Chikungunya viruses. The data obtained here indicate the co-circulation and coinfection by Dengue and Chikungunya viruses in the Tocantins state. These results highlight the importance of monitoring the circulation of these arboviruses for the development of health actions that aim their prevention and combat, as well as their clinical and therapeutic management.

Asunto(s)

Arbovirus , Fiebre Chikungunya , Coinfección , Dengue , Reacción en Cadena de la Polimerasa Multiplex , Humanos , Brasil/epidemiología , Fiebre Chikungunya/diagnóstico , Dengue/diagnóstico , Coinfección/virología , Arbovirus/genética , Arbovirus/aislamiento & purificación , Adulto , Femenino , Masculino , Infección por el Virus Zika/diagnóstico , Adulto Joven , Persona de Mediana Edad , Adolescente , Niño , Reacción en Cadena en Tiempo Real de la Polimerasa , Infecciones por Arbovirus/virología , Infecciones por Arbovirus/diagnóstico , Preescolar , Virus del Dengue/genética , Virus del Dengue/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Virus Chikungunya/genética , Virus Chikungunya/aislamiento & purificación

RESUMEN

Antibiotics and herbicides are contaminants of emerging concern in aquatic environments. Lake Villarrica is a relevant freshwater body in Chile and was recently designated a 'saturated nutrient zone'. Here, we investigated the occurrence of multiple antibiotic resistance (MAR) and herbicide catabolic profiles among bacteria present in the surface sediments of Lake Villarrica. The occurrence of antibiotic-resistant genes (ARGs; blaTEM, catA and tetM) and herbicide-catabolic genes (HCGs; phnJ and atzA) was investigated by qPCR. Subsequently, the presence of culturable bacteria with multiple resistance to amoxicillin (AMX), chloramphenicol (CHL) and oxytetracycline (OXT) was studied. Forty-six culturable MAR (AMX + CHL + OXT) strains were isolated and characterized with respect to their resistance to 11 antibiotics by using a disc diffusion assay and testing their ability to use herbicides as a nutrient source. qPCR analyses revealed that ARGs and HCGs were present in all sediment samples (101 to 103 gene copies g-1), with significant (P ≤ 0.05) higher values in sites near Villarrica city and cattle pastures. The plate method was used to recover MAR isolates from sediment (103-106 CFU g-1), and most of the 46 isolates also showed resistance to oxacillin (100%), cefotaxime (83%), erythromycin (96%) and vancomycin (93%). Additionally, 54 and 57% of the MAR isolates were able to grow on agar supplemented (50 mg L-1) with atrazine and glyphosate as nutrient sources, respectively. Most of the MAR isolates were taxonomically close to Pseudomonas (76.1%) and Pantoea (17.4%), particularly those isolated from urbanized sites (Pucón city). This study shows the presence of MAR bacteria with herbicide catabolic activity in sediments, which is valuable for conservation strategies and risk assessments of Lake Villarrica. However, major integrative studies on sediments as reservoirs or on the fate of MAR strains and traces of antibiotics and herbicides as a result of anthropic pressure are still needed.

Asunto(s)

Antibacterianos , Bacterias , Sedimentos Geológicos , Herbicidas , Lagos , Contaminantes Químicos del Agua , Herbicidas/farmacología , Lagos/microbiología , Sedimentos Geológicos/microbiología , Bacterias/genética , Bacterias/efectos de los fármacos , Bacterias/aislamiento & purificación , Antibacterianos/farmacología , Chile , Monitoreo del Ambiente , Farmacorresistencia Bacteriana Múltiple/genética

RESUMEN

Pseudomonas aeruginosa is one of the most important nosocomial pathogens that possess the ability to produce multiple antibiotic resistance and virulence factors. Elastase B (LasB) is the major factor implicated in tissue invasion and damage during P. aeruginosa infections, whose synthesis is regulated by the quorum sensing (QS) system. Anti-virulence approach is now considered as potential therapeutic alternative and/or adjuvant to current antibiotics' failure. The aim of this study is primarily to find out the impact of the efflux pump inhibitor (EPI) phenylalanine arginyl ß-naphthylamide (PAßN) on the production of elastase B and the gene expression of lasI quorum sensing and lasB virulence factor in clinical isolates of P. aeruginosa. Five P. aeruginosa isolates recovered from patients with respiratory tract infections were examined in this study. Antimicrobial susceptibility of isolates was performed by the disk agar diffusion method. Effect of the PAßN on imipenem susceptibility, bacterial viability, and elastase production was evaluated. The expression of lasB and lasI genes was measured by quantitative real-time PCR in the presence of PAßN. All isolates were identified as multidrug-resistant (MDR) and showed resistance to carbapenem (MIC = 64-256 µg/mL). Susceptibility of isolates to imipenem was highly increased in the presence of efflux inhibitor. PAßN significantly reduced elastase activity in three isolates tested without affecting bacterial growth. In addition, the relative expression of both lasB and lasI genes was diminished in all isolates in the presence of inhibitor. Efflux inhibition by using the EPI PAßN could be a potential target for controlling the P. aeruginosa virulence and pathogenesis. Furthermore, impairment of drug efflux by PAßN indicates its capability to be used as antimicrobial adjuvant that can decrease the resistance and lower the effective doses of current drugs.

Asunto(s)

Antibacterianos , Proteínas Bacterianas , Dipéptidos , Imipenem , Pruebas de Sensibilidad Microbiana , Elastasa Pancreática , Infecciones por Pseudomonas , Pseudomonas aeruginosa , Percepción de Quorum , Factores de Virulencia , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/aislamiento & purificación , Percepción de Quorum/efectos de los fármacos , Factores de Virulencia/genética , Factores de Virulencia/metabolismo , Elastasa Pancreática/genética , Elastasa Pancreática/metabolismo , Antibacterianos/farmacología , Humanos , Infecciones por Pseudomonas/microbiología , Dipéptidos/farmacología , Imipenem/farmacología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Farmacorresistencia Bacteriana , Metaloendopeptidasas

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'Candidatus Phytoplasma brasiliense' (CPB) is a phytoplasma originally discovered in South America and is known to infect a wide variety of economically important crops. It is most prevalent in Hibiscus spp., where it causes witches broom symptoms, and papaya, where it causes bunchy top. Recently, CPB was documented for the first time in North America in a new host, globe sedge. In this study, two quantitative PCR assays are developed: one using high-resolution melt curve analysis (HRMA) based on the secA gene and the other a TaqMan assay based on the dnaK gene. The secA/HRMA and dnaK/TaqMan assay successfully amplified two of the three isolates of CPB. Both assays were screened against available isolates of 16SrI, 16SrII, and 16SrIV phytoplasmas. The secA/HRMA assay failed to amplify 16SrI and 16SrIV phytoplasmas but successfully amplified 16SrII phytoplasmas. The resulting melting point (Tm) products of CPB and 16SrII phytoplasmas displayed a difference of 0.5°C, easily distinguishing them by melt curves. The dnaK/TaqMan assay failed to amplify all non-CPB phytoplasma isolates in the study. The development of these assays provides a valuable tool that will significantly improve monitoring programs in Florida and will aid in developing a better fundamental understanding of the epidemiology of this phytoplasma.

Asunto(s)

Proteínas Bacterianas , Phytoplasma , Enfermedades de las Plantas , Phytoplasma/genética , Phytoplasma/aislamiento & purificación , Phytoplasma/clasificación , Enfermedades de las Plantas/microbiología , Proteínas Bacterianas/genética , ADN Bacteriano/genética , Hibiscus/microbiología , Adenosina Trifosfatasas/genética , Canales de Translocación SEC/genética , Proteína SecA , Proteínas de Transporte de Membrana/genética , ARN Ribosómico 16S/genética , Carica/microbiología , Reacción en Cadena de la Polimerasa/métodos

RESUMEN

This manuscript presents a paired dataset with experimental holograms and their corresponding reconstructed phase maps of human red blood cells (RBCs). The holographic images were recorded using an off-axis telecentric Digital Holographic Microscope (DHM). The imaging system consists of a 40 × /0.65NA infinity-corrected microscope objective (MO) lens and a tube lens (TL) with a focal distance of 200 mm, recording diffraction-limited holograms. A CMOS camera with dimensions of 1920 × 1200 pixels and a pixel pitch of 5.86 µm was located at the back focal plane of the TL lens, capturing image-plane holograms. The off-axis, telecentric, and diffraction-limited DHM system guarantees accurate quantitative phase maps. Initially comprising 300 holograms, the dataset was augmented to 36,864 instances, enabling the investigation (i.e., training and testing) of learning-based models to reconstruct aberration-free phase images from raw holograms. This dataset facilitates the training and testing of end-to-end models for quantitative phase imaging using DHM systems operating at the telecentric regime and non-telecentric DHM systems where the spherical wavefront has been compensated physically. In other words, this dataset holds promise for advancing investigations in digital holographic microscopy and computational imaging.