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Climate change is contributing to biodiversity redistributions and species declines. However, cooler microclimate conditions provided by old-growth forest structures compared with surrounding open or younger forests have been hypothesized to provide thermal refugia for species that are sensitive to climate warming and dampen the negative effects of warming on population trends of animals (i.e., the microclimate buffering hypothesis). In addition to thermal refugia, the compositional and structural diversity of old-growth forest vegetation itself may provide resources to species that are less available in forests with simpler structure (i.e., the insurance hypothesis). We used 8 years of breeding bird abundance data from a forested watershed, accompanied with sub-canopy temperature data, and ground- and LiDAR-based vegetation data to test these hypotheses and identify factors influencing bird population changes from 2011 to 2018. After accounting for imperfect detection, we found that for 5 of 20 bird species analyzed, abundance trends tended to be less negative or neutral at sites with cooler microclimates, which supports the microclimate buffering hypothesis. Negative effects of warming on two species were also reduced in locations with greater forest compositional diversity supporting the insurance hypothesis. We provide the first empirical evidence that complex forest structure and vegetation diversity confer microclimatic advantages to some animal populations in the face of climate change. Conservation of old-growth forests, or their characteristics in managed forests, could help slow the negative effects of climate warming on some breeding bird populations via microclimate buffering and possibly insurance effects.
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Florestas , Microclima , Animais , Biodiversidade , Aves , Mudança Climática , Ecossistema , ÁrvoresRESUMO
DNA Holliday junction (HJ) is a four-way stranded DNA intermediate that formed in replication fork regression, homology-dependent repair and mitosis, performing a significant role in genomic stability. Failure to remove HJ can induce an acceptable replication fork stalling and DNA damage in normal cells, leading to a serious chromosomal aberration and even cell death in HJ nuclease-deficient tumor cells. Thus, HJ is becoming an attractive target in cancer therapy. However, the development of HJ-targeting ligand faces great challenges because of flexile cavities on the center of HJs. This review introduces the discovery history of HJ, elucidates the formation and dissociation procedures of HJ in corresponding bio-events, emphasizes the importance of prompt HJ-removing in genome stability, and summarizes recent advances in HJ-based ligand discovery. Our review indicate that target HJ is a promising approach in oncotherapy.
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DNA Cruciforme , DNA , DNA/metabolismo , Replicação do DNA , DNA Cruciforme/genética , Instabilidade Genômica , Humanos , LigantesRESUMO
Three new tuliposides H-J (1-3) and 11 known compounds were obtained from the methanolic extracts of the bulbs of Amana edulis for the first time. Their structures were elucidated by NMR, MS, and IR spectroscopic data, optical rotation, and Mosher's method. The melanogenesis properties of all the isolates were evaluated in B16 melanoma cells. Consequently, tributyl citrate (9) had anti-melanogenesis activity but was cytotoxic toward B16. (+)-Pyroglutamic acid (4), (+)-butyl 5-oxopyrrolidine-2-carboxylate (6), (-)-3-hydroxy-2-methylbutyrolactone (10), and 5-(hydroxymethyl)furfural (12) had increased melanin productions and tyrosinase activities. Those active components could be further studied as the candidates against melanoma and vitiligo for skin diseases or whitening/hypopigmentation for hair.
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Glucosídeos/farmacologia , Liliaceae/química , Melanoma Experimental/tratamento farmacológico , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/farmacologia , Animais , Melaninas/metabolismo , Camundongos , Monofenol Mono-Oxigenase/metabolismo , Células Tumorais CultivadasRESUMO
Species of eucalypts are commonly cultivated for solid wood and pulp products. The expansion of commercially managed eucalypt plantations has chiefly been driven by their rapid growth and suitability for propagation across a very wide variety of sites and climatic conditions. Infection of foliar fungal pathogens of eucalypts is resulting in increasingly negative impacts on commercial forest industries globally. To assist in evaluating this threat, the present study provides a global perspective on foliar pathogens of eucalypts. We treat 110 different genera including species associated with foliar disease symptoms of these hosts. The vast majority of these fungi have been grown in axenic culture, and subjected to DNA sequence analysis, resolving their phylogeny. During the course of this study several new genera and species were encountered, and these are described. New genera include: Lembosiniella (L. eucalyptorum on E. dunnii, Australia), Neosonderhenia (N. eucalypti on E. costata, Australia), Neothyriopsis (N. sphaerospora on E. camaldulensis, South Africa), Neotrichosphaeria (N. eucalypticola on E. deglupta, Australia), Nothotrimmatostroma (N. bifarium on E. dalrympleana, Australia), Nowamyces (incl. Nowamycetaceae fam. nov., N. globulus on E. globulus, Australia), and Walkaminomyces (W. medusae on E. alba, Australia). New species include (all from Australia): Disculoides fraxinoides on E. fraxinoides, Elsinoe piperitae on E. piperita, Fusculina regnans on E. regnans, Marthamyces johnstonii on E. dunnii, Neofusicoccum corticosae on E. corticosa, Neotrimmatostroma dalrympleanae on E. dalrympleana, Nowamyces piperitae on E. piperita, Phaeothyriolum dunnii on E. dunnii, Pseudophloeospora eucalyptigena on E. obliqua, Pseudophloeospora jollyi on Eucalyptus sp., Quambalaria tasmaniae on Eucalyptus sp., Q. rugosae on E. rugosa, Sonderhenia radiata on E. radiata, Teratosphaeria pseudonubilosa on E. globulus and Thyrinula dunnii on E. dunnii. A new name is also proposed for Heteroconium eucalypti as Thyrinula uruguayensis on E. dunnii, Uruguay. Although many of these genera and species are commonly associated with disease problems, several appear to be opportunists developing on stressed or dying tissues. For the majority of these fungi, pathogenicity remains to be determined. This represents an important goal for forest pathologists and biologists in the future. Consequently, this study will promote renewed interest in foliar pathogens of eucalypts, leading to investigations that will provide an improved understanding of the biology of these fungi.
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Species of Sporocadaceae are endophytic, plant pathogenic or saprobic, and associated with a wide range of host plants. Recent molecular studies that have attempted to address familial and generic boundaries of fungi belonging to Sporocadaceae were based on a limited number of samples and DNA loci. The taxonomy of this group of fungi is therefore still not fully resolved. The aim of the present study is to provide a natural classification for the Sporocadaceae based on multi-locus phylogenetic analyses, using LSU, ITS, tef-1α, tub2 and rpb2 loci, in combination with morphological data. A total of 30 well-supported monophyletic clades in Sporocadaceae are recognised, representing 23 known and seven new genera. Typifications are proposed for the type species of five genera (Diploceras, Discosia, Monochaetia, Sporocadus and Truncatella) to stabilise the application of these names. Furthermore, Neotruncatella and Dyrithiopsis are synonymised under Hymenopleella, and the generic circumscriptions of Diploceras, Disaeta, Hymenopleella, Monochaetia, Morinia, Pseudopestalotiopsis, Sarcostroma, Seimatosporium, Synnemapestaloides and Truncatella are emended. A total of 51 new species, one nomina nova and 15 combinations are introduced.
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Homologous recombination (HR) is a complex biological process and is central to meiosis and for repair of DNA double-strand breaks. Although the HR process has been the subject of intensive study for more than three decades, the complex protein-protein and protein-DNA interactions during HR present a significant challenge for determining the molecular mechanism(s) of the process. This knowledge gap is largely because of the dynamic interactions between HR proteins and DNA which is difficult to capture by routine biochemical or structural biology methods. In recent years, single-molecule fluorescence microscopy has been a popular method in the field of HR to visualize these complex and dynamic interactions at high spatiotemporal resolution, revealing mechanistic insights of the process. In this review, we describe recent efforts that employ single-molecule fluorescence microscopy to investigate protein-protein and protein-DNA interactions operating on three key DNA-substrates: single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), and four-way DNA called Holliday junction (HJ). We also outline the technological advances and several key insights revealed by these studies in terms of protein assembly on these DNA substrates and highlight the foreseeable promise of single-molecule fluorescence microscopy in advancing our understanding of homologous recombination.
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Recombinação Homóloga/genética , Microscopia de Fluorescência/métodos , Imagem Individual de Molécula/métodos , Animais , DNA/genética , DNA Cruciforme/metabolismo , Humanos , Ligação ProteicaRESUMO
Forest plays a significant role in the global carbon budget and ecological processes. The precise mapping of forest cover can help significantly reduce uncertainties in the estimation of terrestrial carbon balance. A reliable and operational method is necessary for a rapid regional forest mapping. In this study, the goal relies on mapping forest and subcategories in Northeast China through the use of high spatio-temporal resolution HJ-1 imagery and time series vegetation indices within the context of an object-based image analysis and decision tree classification. Multi-temporal HJ-1 images obtained in a single year provide an opportunity to acquire phenology information. By analyzing the difference of spectral and phenology information between forest and non-forest, forest subcategories, decision trees using threshold values were finally proposed. The resultant forest map has a high overall accuracy of 0.91 ± 0.01 with a 95% confidence interval, based on the validation using ground truth data from field surveys. The forest map extracted from HJ-1 imagery was compared with two existing global land cover datasets: GlobCover 2009 and MCD12Q1 2009. The HJ-1-based forest area is larger than that of MCD12Q1 and GlobCover and more closely resembles the national statistics data on forest area, which accounts for more than 40% of the total area of the Northeast China. The spatial disagreement primarily occurs in the northern part of the Daxing'an Mountains, Sanjiang Plain and the southwestern part of the Songliao Plain. The compared result also indicated that the forest subcategories information from global land cover products may introduce large uncertainties for ecological modeling and these should be cautiously used in various ecological models. Given the higher spatial and temporal resolution, HJ-1-based forest products could be very useful as input to biogeochemical models (particularly carbon cycle models) that require accurate and updated estimates of forest area and type.
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Crop yield estimation is important for formulating informed regional and national food trade policies. The introduction of remote sensing in agricultural monitoring makes accurate estimation of regional crop yields possible. However, remote sensing images and crop distribution maps with coarse spatial resolution usually cause inaccuracy in yield estimation due to the existence of mixed pixels. This study aimed to estimate the annual yields of maize and sunflower in Hetao Irrigation District in North China using 30 m spatial resolution HJ-1A/1B CCD images and high accuracy multi-year crop distribution maps. The Normalized Difference Vegetation Index (NDVI) time series obtained from HJ-1A/1B CCD images was fitted with an asymmetric logistic curve to calculate daily NDVI and phenological characteristics. Eight random forest (RF) models using different predictors were developed for maize and sunflower yield estimation, respectively, where predictors of each model were a combination of NDVI series and/or phenological characteristics. We calibrated all RF models with measured crop yields at sampling points in two years (2014 and 2015), and validated the RF models with statistical yields of four counties in six years. Results showed that the optimal model for maize yield estimation was the model using NDVI series from the 120th to the 210th day in a year with 10 days' interval as predictors, while that for sunflower was the model using the combination of three NDVI characteristics, three phenological characteristics, and two curve parameters as predictors. The selected RF models could estimate multi-year regional crop yields accurately, with the average values of root-mean-square error and the relative error of 0.75 t/ha and 6.1% for maize, and 0.40 t/ha and 10.1% for sunflower, respectively. Moreover, the yields of maize and sunflower can be estimated fairly well with NDVI series 50 days before crop harvest, which implicated the possibility of crop yield forecast before harvest.
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Agricultura/tendências , Tecnologia de Sensoriamento Remoto/tendências , Comunicações Via Satélite/tendências , Zea mays/crescimento & desenvolvimento , China , Humanos , Estações do AnoRESUMO
A recent taxonomic revision of Microascaceae with an emphasis on synnematous fungi enabled re-identification of previously isolated indoor strains of Cephalotrichum. All available Cephalotrichum strains from the culture collection of the Westerdijk Institute were studied, 20 originating from the built environment. Phylogenetic relationships were inferred from DNA sequence data from the internal transcribed spacer 1 and 2 and intervening 5.8S nrDNA (ITS), and parts of ß-tubulin (tub2) and translation elongation factor 1-α (tef1) genes. Additionally, herbarium material of 14 Cephalotrichum species described from soil in China was studied, and the taxonomy of C. album, not considered in recent revisions, was reevaluated. Sixteen phylogenetic species in Cephalotrichum are distinguished, five described as new species: C. domesticum, C. lignatile, C. telluricum, C. tenuissimum and C. transvaalense. Five Cephalotrichum species occur in the built environment: C. domesticum, C. gorgonifer (formerly known as Trichurus spiralis), C. microsporum, C. purpureofuscum, and C. verrucisporum. Based on the number of isolates, C. gorgonifer (nine strains) is the most common indoor species. The study of the Chinese herbarium material resulted in the acceptance of three additional Cephalotrichum species: C. casteneum, C. ellipsoideum, and C. spirale. Four species are considered nomena dubia (C. cylindrosporum, C. macrosporum, C. ovoideum, and C. robustum), five are placed in synonymy with other Cephalotrichum species (C. acutisporum, C. inflatum, C. longicollum, C. oblongum, C. terricola) and one species, C. verrucipes, is probably a synonym of Penicillium clavigerum. Cephalotrichum columnare, former Doratomyces columnaris, is transferred to Kernia. Cephalotrichum album, formerly known as Doratomyces putredinis, is transferred to Acaulium and redescribed.
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An effective method to monitor heavy metal stress in crops is of critical importance to assure agricultural production and food security. Phenology, as a sensitive indicator of environmental change, can respond to heavy metal stress in crops and remote sensing is an effective method to detect plant phenological changes. This study focused on identifying the rice phenological differences under varied heavy metal stress using EVI (enhanced vegetation index) time-series, which was obtained from HJ-1A/B CCD images and fitted with asymmetric Gaussian model functions. We extracted three phenological periods using first derivative analysis: the tillering period, heading period, and maturation period; and constructed two kinds of metrics with phenological characteristics: date-intervals and time-integrated EVI, to explore the rice phenological differences under mild and severe stress levels. Results indicated that under severe stress the values of the metrics for presenting rice phenological differences in the experimental areas of heavy metal stress were smaller than the ones under mild stress. This finding represents a new method for monitoring heavy metal contamination through rice phenology.
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Oryza , Agricultura , Produtos Agrícolas , Metais PesadosRESUMO
The taxonomy of the synnematous genera Cephalotrichum, Doratomyces and Trichurus, and other related genera Gamsia, Wardomyces and Wardomycopsis, has been controversial and relies mainly on morphological criteria. These are microascaceous saprobic fungi mostly found in air and soil and with a worldwide distribution. In order to clarify their taxonomy and to delineate generic boundaries within the Microascaceae, we studied 57 isolates that include clinical, environmental and all the available ex-type strains of a large set of species by means of morphological, physiological and molecular phylogenetic analyses using DNA sequence data of four loci (the ITS region, and fragments of rDNA LSU, translation elongation factor 1α and ß-tubulin). The results demonstrate that Cephalotrichum, Doratomyces and Trichurus are congeneric and the genus Cephalotrichum is accepted here with Echinobotryum as a further synonym. The genera Acaulium and Fairmania, typified by A. albonigrescens and F. singularis, respectively, are distinct from Microascus and Scopulariopsis, Gamsia is distinct from Wardomyces, and Wardomycopsis is confirmed as a separate genus in the Microascaceae. Two new species of Cephalotrichum are described as C. brevistipitatum and C. hinnuleum. Nine new combinations are proposed, i.e. Acaulium acremonium, A. caviariforme, Cephalotrichum asperulum, C. columnare, C. cylindricum, C. dendrocephalum, C. gorgonifer, Gamsia columbina and Wardomyces giganteus. A neotype is designed for C. stemonitis. Lectotypes and epitypes are designated for A. acremonium, A. albonigrescens, C. gorgonifer, C. nanum and W. anomalus. Cephalotrichum cylindricum, C. microsporum, F. singularis and Gamsia columbina are also epitypified with new specimens. Descriptions of the phenotypic features and dichotomous keys for identification are provided for accepted species in the different genera.
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Accurate and timely mapping of paddy rice is vital for food security and environmental sustainability. This study evaluates the utility of temporal features extracted from coarse resolution data for object-based paddy rice classification of fine resolution data. The coarse resolution vegetation index data is first fused with the fine resolution data to generate the time series fine resolution data. Temporal features are extracted from the fused data and added with the multi-spectral data to improve the classification accuracy. Temporal features provided the crop growth information, while multi-spectral data provided the pattern variation of paddy rice. The achieved overall classification accuracy and kappa coefficient were 84.37% and 0.68, respectively. The results indicate that the use of temporal features improved the overall classification accuracy of a single-date multi-spectral image by 18.75% from 65.62% to 84.37%. The minimum sensitivity (MS) of the paddy rice classification has also been improved. The comparison showed that the mapped paddy area was analogous to the agricultural statistics at the district level. This work also highlighted the importance of feature selection to achieve higher classification accuracies. These results demonstrate the potential of the combined use of temporal and spectral features for accurate paddy rice classification.
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Monitoramento Ambiental/métodos , Oryza/crescimento & desenvolvimento , Agricultura , Algoritmos , Produtos Agrícolas/crescimento & desenvolvimentoRESUMO
Tomatoes have been described as a functional food because of their particular composition of different bioactive compounds. In this study, the proximate composition, minerals and trace elements, and antioxidant compounds were determined in two tomato cultivars (Mariana and Dunkan) that were grown in Gran Canaria (Spain) either conventionally or hydroponically. Although compositional data of this type require being subjected to the specific statistical techniques of compositional analysis, this approach has not usually been considered in this context. In the present case, a compositional Mann-Whitney U test of the data showed significant differences for each factor (cultivar and cultivation system) in several of the compositional variables studied. For the differences between cultivars, these parameters were the protein, Mg, lycopene, ascorbic acid, citric acid, and fumaric acid contents. For the differences between cultivation systems, they were mainly those of the mineral and trace elements group. Although one-year data are insufficient to make clear relationship among compounds because more repetitions in several localities and years are necessary, the compositional HJ-biplot (in which the links provide estimates of the linear relationship among variables) results agreed with other scientific results about linear relationship among some compounds analyzed.
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Antioxidantes/análise , Carotenoides/análise , Frutas/química , Magnésio/análise , Proteínas de Plantas/análise , Solanum lycopersicum/química , Antioxidantes/metabolismo , Ácido Ascórbico/análise , Ácido Ascórbico/metabolismo , Carotenoides/metabolismo , Cátions Bivalentes , Ácido Cítrico/análise , Ácido Cítrico/metabolismo , Gráficos por Computador , Frutas/metabolismo , Fumaratos/análise , Fumaratos/metabolismo , Hidroponia , Solanum lycopersicum/classificação , Solanum lycopersicum/metabolismo , Magnésio/metabolismo , Melhoramento Vegetal , Proteínas de Plantas/metabolismo , Especificidade da Espécie , Estatísticas não ParamétricasRESUMO
Lantibiotic bovicin HJ50 is produced by Streptococcus bovis HJ50 and acts as the extracellular signal to autoregulate its own biosynthesis through BovK/R two-component system. Bovicin HJ50 shows a linear N-terminal and glubolar C-terminal structure, and the sensor histidine kinase BovK contains eight transmembrane segments lacking any extensive surface-exposed sensory domain. The signal recognition mechanism between bovicin HJ50 and BovK is still unknown. We performed saturated alanine scanning mutagenesis and other amino acid substitutions on bovicin HJ50 using a semi-in vitro biosynthesis. Results of the mutants inducing activities indicated that several charged and hydrophobic amino acids in ring B of bovicin HJ50, as well as two glycines were key residues to recognize BovK. Circular dichroism analyses indicated that both glycines contributed to bovicin HJ50 structural changes in the membrane. Biotin-labeled bovicin HJ50 could interact with the N-terminal sensor of BovK, and several charged residues and a conserved hydrophobic region in the N-terminal portion of BovK sensor domain were important for interacting with the signal bovicin HJ50. By combining the results, we suggested a mechanism of bovicin HJ50 recognizing and activating BovK mainly through electrostatic and hydrophobic interactions.
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Proteínas de Bactérias/metabolismo , Bacteriocinas/metabolismo , Proteínas Quinases/metabolismo , Transdução de Sinais/fisiologia , Streptococcus bovis/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Bacteriocinas/química , Bacteriocinas/genética , Histidina Quinase , Mutação , Proteínas Quinases/química , Proteínas Quinases/genética , Estrutura Terciária de Proteína , Streptococcus bovis/química , Streptococcus bovis/genéticaRESUMO
The ascomycete family Nectriaceae (Hypocreales) includes numerous important plant and human pathogens, as well as several species used extensively in industrial and commercial applications as biodegraders and biocontrol agents. Members of the family are unified by phenotypic characters such as uniloculate ascomata that are yellow, orange-red to purple, and with phialidic asexual morphs. The generic concepts in Nectriaceae are poorly defined, since DNA sequence data have not been available for many of these genera. To address this issue we performed a multi-gene phylogenetic analysis using partial sequences for the 28S large subunit (LSU) nrDNA, the internal transcribed spacer region and intervening 5.8S nrRNA gene (ITS), the large subunit of the ATP citrate lyase (acl1), the RNA polymerase II largest subunit (rpb1), RNA polymerase II second largest subunit (rpb2), α-actin (act), ß-tubulin (tub2), calmodulin (cmdA), histone H3 (his3), and translation elongation factor 1-alpha (tef1) gene regions for available type and authentic strains representing known genera in Nectriaceae, including several genera for which no sequence data were previously available. Supported by morphological observations, the data resolved 47 genera in the Nectriaceae. We re-evaluated the status of several genera, which resulted in the introduction of six new genera to accommodate species that were initially classified based solely on morphological characters. Several generic names are proposed for synonymy based on the abolishment of dual nomenclature. Additionally, a new family is introduced for two genera that were previously accommodated in the Nectriaceae.
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BACKGROUND: Pre-school age is important for developing healthy attitudes towards physical activity (PA). However, research on pre-schoolers' pedometer-determined PA is limited. AIM: To describe pre-schoolers' ambulatory activity; investigate step count differences in respect to Body Mass Index (BMI) categories and examine the prevalence of obesity. SUBJECTS AND METHODS: Pre-school aged children (n = 250; 5.5 ± 0.4 years) from Komotini (Greece) wore Omron HJ-720IT-E2 pedometers for 10 consecutive days. Height and weight were measured and BMI was calculated. RESULTS: Three-way repeated measures ANOVAs revealed that children performed more steps on weekdays than during weekends (p < 0.001) and during leisure time than school (p < 0.001). Significant differences appeared between normal and obese children's counts on weekdays (p < 0.001), weekend days (p < 0.05), during school (p < 0.001), after school (p < 0.005) and in weekly steps (p < 0.005). No gender differences were detected. Moreover, according to a sample t-test analysis, children's daily steps were significantly different from the 10,000 steps/day guideline, while obesity prevalence was 15.6%. CONCLUSION: School-based ambulatory activity is lower than after school ambulatory activity, independent of BMI-category and gender, although obese children demonstrated fewer steps. Taking into account the high rate of both the obesity prevalence and children not meeting the 10,000 steps/day guideline, the need for preventive policies becomes obvious.
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Atividade Motora , Obesidade/epidemiologia , Acelerometria , Criança , Pré-Escolar , Feminino , Grécia/epidemiologia , Humanos , Masculino , Obesidade/etiologia , PrevalênciaRESUMO
BACKGROUND: Keratomycosis is a form of infectious keratitis, an infection of the cornea, which is caused by fungi. This disease is a leading cause of ocular morbidity globally with at least 60 % of the affected individuals becoming monocularly blind. OBJECTIVE: This bibliometric analysis aimed to comprehensively assess the existing body of literature, providing insights of the evolution of keratomycosis research by identifying key themes and research gaps. METHODS: This work used the modeling method Latent Dirichlet Allocation (LDA) to identify and interpret scientific information on topics concerning existing categories in a set of documents. The HJ-Biplot method was also used to determine the relationship between the analyzed topics, taking into consideration the years under study. RESULTS: This bibliometric analysis was performed on a total of 2,599 scientific articles published between 1992 and 2022. The five leading countries with more scientific production and citations on keratomycosis were The United States of America, followed by India, China, United Kingdom and Australia. The top five topics studied were Case Reports and Corneal Infections, which exhibited a decreasing trend; followed by Penetrating Keratoplasty and Corneal Surgery, Ocular Effects of Antifungal Drugs, Gene Expression and Inflammatory Response in the Cornea and Patient Data which have been increasing throughout the years. However Filamentous Fungi and Specific Pathogens, and Antifungal Therapies research has been decreasing in trend. CONCLUSION: Additional investigation into innovative antifungal drug therapies is crucial for proactively tackling the potential future resistance to antifungal agents in scientific writing.
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Bibliometria , Infecções Oculares Fúngicas , Ceratite , Humanos , Ceratite/microbiologia , Infecções Oculares Fúngicas/microbiologia , Antifúngicos/uso terapêutico , Saúde Global , Fungos/classificação , Fungos/isolamento & purificação , Córnea/microbiologiaRESUMO
ETHNOPHARMACOLOGY RELEVANCE: Zanthoxylum bungeanum Maxim. (Z. bungeanum), a member of the Rutaceae family, has a rich history of traditional use in Asia for treating arthritis and toothache conditions. As characteristic chemical components, numerous kinds of alkaloids have been extracted from plants and their diverse biological activities have been reported. However, research on the isoquinoline alkaloid, a specific type of alkaloids, in Z. bungeanum was scarce. AIM OF THE STUDY: The study aimed to isolate a novel isoquinoline alkaloid from Z. bungeanum and explore its pharmacological activity in vitro and analgesic activity in vivo. MATERIALS AND METHODS: Isoquinoline alkaloid isolation and identification from Z. bungeanum were conducted using chromatographic and spectroscopic methods. The whole-cell patch-clamp technique was applied to assess its impact on neuronal excitability, and endogenous voltage-gated potassium (Kv) and sodium (Nav) currents in acutely isolated mouse small-diameter dorsal root ganglion (DRG) neurons. Its inhibitory impacts on channels were further validated with HEK293 cells stably expressing Nav1.7 and Nav1.8, and Chinese hamster ovary (CHO) cells transiently expressing Kv2.1. The formalin inflammatory pain model was utilized to evaluate the potential analgesic activity in vivo. RESULTS: A novel isoquinoline alkaloid named HJ-69 (N-13-(3-methoxyprop-1-yl)rutaecarpine) was isolated and identified from Z. bungeanum for the first time. HJ-69 significantly suppressed the firing frequency and amplitudes of action potentials in DRG neurons. Consistently, it state-dependently inhibited endogenous Nav currents of DRG neurons, with half maximal inhibitory concentration (IC50) values of 13.06 ± 2.06 µM and 30.19 ± 2.07 µM for the inactivated and resting states, respectively. HJ-69 significantly suppressed potassium currents in DRG neurons, which notably inhibited the delayed rectifier potassium (IK) currents (IC50 = 6.95 ± 1.29 µM) and slightly affected the transient outward potassium (IA) currents (IC50 = 523.50 ± 39.16 µM). Furtherly, HJ-69 exhibited similar potencies on heterologously expressed Nav1.7, Nav1.8, and Kv2.1 channels, which correspondingly represent the main components in neurons. Notably, intraperitoneal administration of 30 mg/kg and 100 mg/kg HJ-69 significantly alleviated pain behaviors in the mouse inflammatory pain model induced by formalin. CONCLUSION: The study concluded that HJ-69 is a novel and active isoquinoline alkaloid, and the inhibition of Nav and Kv channels contributes to its analgesic activity. HJ-69 may be a promising prototype for future analgesic drug discovery based on the isoquinoline alkaloid.
Assuntos
Analgésicos , Gânglios Espinais , Dor , Zanthoxylum , Animais , Zanthoxylum/química , Humanos , Células HEK293 , Analgésicos/farmacologia , Analgésicos/química , Analgésicos/isolamento & purificação , Analgésicos/uso terapêutico , Gânglios Espinais/efeitos dos fármacos , Gânglios Espinais/metabolismo , Camundongos , Masculino , Dor/tratamento farmacológico , Isoquinolinas/farmacologia , Isoquinolinas/isolamento & purificação , Isoquinolinas/química , Alcaloides/farmacologia , Alcaloides/isolamento & purificação , Alcaloides/química , Alcaloides/uso terapêutico , Bloqueadores dos Canais de Potássio/farmacologia , Canal de Sódio Disparado por Voltagem NAV1.7/metabolismo , Inflamação/tratamento farmacológico , Bloqueadores do Canal de Sódio Disparado por Voltagem/farmacologia , Bloqueadores do Canal de Sódio Disparado por Voltagem/isolamento & purificação , Canais de Potássio de Abertura Dependente da Tensão da Membrana/metabolismo , Canais de Potássio de Abertura Dependente da Tensão da Membrana/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/química , Camundongos Endogâmicos C57BL , CricetulusRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Urinary tract infections (UTIs) are globally prevalent infectious diseases, predominantly caused by uropathogenic Escherichia coli (UPEC). The misuse of antibiotics has led to the emergence of several drug-resistant strains. Traditional Chinese Medicine (TCM) has its own advantages in the treatment of UTIs. HJ granules is a herbal formula used for the treatment of UTIs. However, its mechanism of action is not clear. AIM OF THE STUDY: The aim of this study was to investigate the therapeutic efficacy and mechanism of action of HJ granules in a rat model of UTI caused by Escherichia coli (E coli) CFT073. MATERIALS AND METHODS: SD rats were selected to establish a rat UTI model by injecting UPEC strain CFT073 into the bladder using the transurethral placement method. HJ granules were administered to rats after modelling and the efficacy of HJ granule was investigated by measuring urinary decanalogue, inflammatory factors in bladder tissue and pathological changes in the bladder after 3d of administration. Expression of sonic hedgehog (SHH), NOD-like receptor thermoprotein domain 3 (NLRP3), apoptosis-associated speck-like protein (ASC) and activation of cysteinyl aspartate specific proteinase-1 (caspase-1) were detected by western blotting and immunofluorescence staining in rat bladder tissue. NLRP3, ASC and caspase-1, a cysteine-containing aspartic protein, were expressed and activated. RESULTS: The results showed that infection of rats with UPEC resulted in increased pH and erythrocytes in bladder irrigation fluid; increased expression of IL-1ß, IL-6 and SHH and decreased expression of IL-10 in bladder tissue; and significant upregulation of the expression of both SHH and NLRP3 inflammasom and significant activation of NLRP3 inflammasom. HJ granules significantly increased the concentration of IL-10 in the bladder, inhibited the expression of SHH and NLRP3 inflammasom in bladder tissue, and suppressed the activation of NLRP3 inflammasom, thereby reducing inflammatory lesions in bladder tissue. CONCLUSION: HJ granules may improve bladder injury and treat UTIs by inhibiting the expression and activation of NLRP3 inflammasom.
Assuntos
Infecções por Escherichia coli , Infecções Urinárias , Escherichia coli Uropatogênica , Ratos , Animais , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Escherichia coli , Interleucina-10 , Proteínas Hedgehog , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/patologia , Ratos Sprague-Dawley , Infecções Urinárias/tratamento farmacológico , Infecções Urinárias/patologia , Caspase 1/metabolismoRESUMO
Video 1Narrated case of an EUS-guided hepaticogastrostomy facilitated by opacification and distention of the left intrahepatic ducts using an existing percutaneous drain tract.