Quantitative real‐time PCR (qPCR) has become one of the most used techniques to measure gene expression. However, normalization of gene expression data against reference genes is essential, although ...these are usually used without any kind of validation. The expression of seven genes was compared in organs of Rhodnius prolixus under diverse conditions, using published software to test gene expression stability. Rp18S and elongation factor 1 (RpEF ‐1) were the most reliable genes for normalization in qPCR when gene expression in different organs was compared. Moreover, both genes were found to be the best references when transcript levels were compared in the posterior midgut of insects infected with Trypanosoma cruzi. Rp18S was also the best reference gene in the fat bodies of unfed and fed insects. By contrast, RpEF‐1 was found to be the best reference gene for comparison between posterior midguts, and RpMIP or RpActin should be used to compare gene expression in the ovaries. Although Rp18S is indicated here as the best reference in most cases, reports from the literature show that it is difficult to find an optimum reference gene. Nevertheless, validation of candidate genes to be taken as references is important when new experimental conditions are tested to avoid incorrect data interpretation.
Science studies have been a field of research for different knowledge areas, and they have been successfully used to analyse the construction of scientific knowledge, practice and dissemination. In ...this study, we aimed to verify how the Zika epidemic has moulded the scientific articles published worldwide by analysing international collaborations and the knowledge landscape through time, as well as research topics and country involvement.
We searched the Web of Science (WoS), Scopus and PubMed for studies published up to 31st December 2018 on Zika using the search terms "zika", "zkv" or "zikv". We analysed the scientific production regarding which countries have published the most, on which topics, as well as country level collaboration. We performed a scientometric analysis of research on Zika focusing on knowledge mapping and the scientific research path over time and space.
We found two well defined research areas divided into three subtopics accounting for six clusters. With regard to country analysis, the USA and Brazil were the countries with the highest numbers of publications on Zika. China entered as a new player focusing on specific research areas. When we took into consideration the epidemics and reported cases, Brazil and France were the leading research countries on related topics. As for international collaboration, the USA followed by England and France stand out as the main hubs. The research areas most published included public health-related topics from 2015 until the very beginning of 2016, followed by an increase in topics related to the clinical aspects of the disease in 2016 and the emergence of laboratory research in 2017/2018.
Mapping the response to Zika, a public health emergency, demonstrated a clear pattern of the participation of countries in the scientific advances. The pattern of knowledge production found in this study represented varying country perspectives, research capacity and interests based first on their level of exposure to the epidemic and second on their financial positions regarding science.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Background/Aim
Childhood dental trauma poses a prevalent concern, and the limited knowledge of lay caregivers toward the needed immediate actions can adversely affect a child's well‐being. This study ...aimed to create and validate an educational comic book for lay individuals, designed to guide immediate actions in dental trauma cases, and assess its effects toward enhancing reader knowledge.
Methodology
The four‐stage study involved: (1) Literature review and script development; (2) Version 1 of the comic book creation (including illustrations, layout, and design) and Flesch Readability Index (FI) calculation; (3) Version 1 validation by expert judges, assessed by the Content Validity Index (CVI); (4) Version 1 refinement based on the judges' feedback, recalculation of FI, and a pilot test (CVI) involving lay individuals who validated the final material (Version 2). The validation process involved 67 participants, including 13 specialist judges and 54 elementary school teachers.
Results
The CVI for Version 1 was calculated at 0.89 (89%), indicating a favorable level of agreement on the evaluated criteria. The FI was determined as 91.1%, indicating a high level of readability. The FI for Version 2 was 92.1% (indicating a very easy level of comprehension). The CVI calculated for the target audience after the pilot test was 0.99 (99%), denoting a substantial agreement among the participants. In assessing the level of knowledge, a significant increase was observed in the rate of correct answers after the comic book was read by lay individuals (p < 0.05), inasmuch as the percentage rose from 50.5% before to 81.2% after reading it.
Conclusion
The educational comic book proved valid in terms of readability, appearance, and content. Furthermore, it had a positive impact on gaining the knowledge needed to take the appropriate actions, thus making it a valuable tool for guiding the lay population in cases of dental trauma.
Amazon rainforest has been subjected to particularly high rates of deforestation caused mainly by the expansion of cattle pasture and agriculture. A commonly observed response to land-use change is a ...negative impact on biodiversity of plant and animal species. However, its effect on the soil microbial community and ecosystem functioning is still poorly understood. Here, we used a DNA metagenomic sequencing approach to investigate the impact of land-use change on soil microbial community composition and its potential functions in three land-use systems (primary forest, pasture and secondary forest) in the Amazon region. In general, the microbial community structure was influenced by changes in soil physicochemical properties. Aluminum and water-holding capacity significantly correlated to overall community structure and most of microbial phyla. Taxonomic changes were followed by potential functional changes in the soil microbial community, with pasture presenting the most distinct profile in comparison with other sites. Although taxonomic structure was very distinct among sites, we observed a recovery of the potential functions in secondary forest after pasture abandonment. Our findings elucidate a significant shift in belowground microbial taxonomic and potential functional diversity following natural forest re-establishment and have implications for ecological restoration programs in tropical and sub-tropical ecosystems.
Purpose
Triple-negative breast cancer (TNBC), an aggressive breast cancer subtype, is genetically heterogeneous which challenges the identification of clinically effective molecular makers. ...Extracellular vesicles (EVs) are key players in the intercellular signaling communication and have been shown to be involved in tumorigenesis. The main goal of this study was to evaluate the role and mechanisms of EVs derived from TNBC cells in modulating proliferation and cytotoxicity to chemotherapeutic agents in non-tumorigenic breast cells (MCF10A).
Methods
EVs were isolated from TNBC cell lines and characterized by nanoparticle tracking analysis, Western blot, and transmission electron microscopy. MCF10A cells were treated with the isolated EVs and evaluated for cell proliferation and cytotoxicity to Docetaxel and Doxorubicin by the MTT and MTS assays, respectively. Gene and miRNA expression profiling was performed in the treated cells to determine expression changes that may be caused by EVs treatment.
Results
MCF10A cells treated with HCC1806-EVs (MCF10A/HCC1806-EVs) showed a significant increase in cell proliferation and resistance to the therapeutic agents tested. No significant effects were observed in the MCF10A cells treated with EVs derived from MDA-MB-231 cells. Gene and miRNA expression profiling revealed 138 genes and 70 miRNAs significantly differentially expressed among the MCF10A/HCC1806-EVs and the untreated MCF10A cells, affecting mostly the PI3K/AKT, MAPK, and HIF1A pathways.
Conclusion
EVs isolated from the HCC1806 TNBC cells are capable of inducing proliferation and drug resistance on the non-tumorigenic MCF10A breast cells, potentially mediated by changes in genes and miRNAs expression associated with cell proliferation, apoptosis, invasion, and migration.
Sulfometuron-methyl is a broad-spectrum herbicide, used throughout Brazil; however, its environmental impacts in biochar (BC) amended soils is not fully understood. Biochar is known to enhance soil ...quality but can also have undesired effects such as altering the bioavailability and behavior of herbicides. Microbial communities can degrade herbicides such as sulfometuron-methyl in soils; however, they are known to be affected by BC. Therefore, it is important to understand the tripartite interaction between these factors. This research aimed to evaluate the sorption-desorption and biodegradation of sulfometuron-methyl in Amazonian soils amended with BC, and to assess the effects of the interactions between BC and sulfometuron-methyl on soil bacterial communities. Soil samples were collected from field plots amended with BC at three doses (0, 40 and 80 t ha−1) applied ten years ago. The herbicide sorption and desorption were evaluated using a batch equilibrium method. Mineralization and biodegradation studies were conducted in microcosms incubated with 14C-sulfometuron-methyl for 80 days. Systematic soil sampling, followed by DNA extraction, quantification (qPCR) and 16S rRNA amplicon sequencing were performed. The presence of BC increased the sorption of the herbicide to the soil by 11% (BC40) and 16% (BC80) compared to unamended soil. The presence of BC also affected the degradation of 14C-sulfometuron-methyl, reducing the mineralization rate and increasing the degradation half-life times (DT50) from 36.67 days in unamended soil to 52.11 and 55.45 days in BC40 and BC80 soils, respectively. The herbicide application altered the bacterial communities, affecting abundance and richness, and changing the taxonomic diversity (i.e., some taxa were promoted and other inhibited). A tripartite interaction was found between BC, the herbicide and soil bacterial communities, suggesting that it is important to consider the environmental impact of soil applied herbicides in biochar amended soils.
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•The effects of sulfometuron-methyl on soil bacteria were explored in biochar-amended soils.•Biochar addition increased herbicide sorption in soils by more than 10%.•Biochar affected herbicide degradation, increasing half-life from 36 to more than 50 days.•The herbicide reduced bacterial richness, especially in the presence of biochar.•Different taxa were promoted/inhibited by the herbicide, depending on the biochar levels.
Lactobacillus acidophilus were encapsulated by complex coacervation followed by transglutaminase crosslinking, aiming to improve the resistance of the microcapsules and improve the protection for ...probiotics. Subsequently, microcapsules were dried by freeze drying. The encapsulation efficiency, morphology, thermal resistance, gastrointestinal simulation and storage stability were analysed for wet and dry forms. The treatments offered high encapsulation efficiency (68.20–97.72%). Transglutaminase maintained the structure rounded, multinucleate and homogeneous distribution of probiotics in the microcapsules. In relation to the thermal resistance, in general, microencapsulation was effective in protecting and crosslinked microcapsules demonstrated greater protection for probiotics, obtaining viable cell counts of up to 10 log CFU g−1, approximately. On exposure to the simulated gastrointestinal tract, microencapsulation coupled to crosslinking demonstrated good results and the dry form was more efficient in the protection and the treatment with greater amount of transglutaminase was highlighted (9.07 log CFU g−1). As for storage, probiotic viability was maintained for up to 60 days in freezing temperature, with counts of up to 9.59 log CFU g−1. The results obtained in the present work are innovative and present a promising alternative for the protection of probiotics and their addition in food products.
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•Transglutaminase maintained the structure of the microcapsules.•Crosslinked microcapsules demonstrated greater protection for probiotics.•Transglutaminase influenced the survival of the microorganism in storage.
Cotton plants are subjected to the attack of several insect pests. In Brazil, the cotton boll weevil, Anthonomus grandis, is the most important cotton pest. The use of insecticidal proteins and gene ...silencing by interference RNA (RNAi) as techniques for insect control are promising strategies, which has been applied in the last few years. For this insect, there are not much available molecular information on databases. Using 454-pyrosequencing methodology, the transcriptome of all developmental stages of the insect pest, A. grandis, was analyzed. The A. grandis transcriptome analysis resulted in more than 500.000 reads and a data set of high quality 20,841 contigs. After sequence assembly and annotation, around 10,600 contigs had at least one BLAST hit against NCBI non-redundant protein database and 65.7% was similar to Tribolium castaneum sequences. A comparison of A. grandis, Drosophila melanogaster and Bombyx mori protein families' data showed higher similarity to dipteran than to lepidopteran sequences. Several contigs of genes encoding proteins involved in RNAi mechanism were found. PAZ Domains sequences extracted from the transcriptome showed high similarity and conservation for the most important functional and structural motifs when compared to PAZ Domains from 5 species. Two SID-like contigs were phylogenetically analyzed and grouped with T. castaneum SID-like proteins. No RdRP gene was found. A contig matching chitin synthase 1 was mined from the transcriptome. dsRNA microinjection of a chitin synthase gene to A. grandis female adults resulted in normal oviposition of unviable eggs and malformed alive larvae that were unable to develop in artificial diet. This is the first study that characterizes the transcriptome of the coleopteran, A. grandis. A new and representative transcriptome database for this insect pest is now available. All data support the state of the art of RNAi mechanism in insects.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The propolis is extensively used in folk medicine in natura or to prepare pharmaceutical formulations since ancient time to improve health or prevent diseases, among them gastrointestinal disorders. ...Aiming to contribute in the scientific validation about the popular use of Brazilian Green propolis (BGP) against gastritis and gastric ulcer, this work evaluated the antiulcer potential of isolated compounds from BGP, three prenylated p-coumaric acid derivatives and two flavonoids, respectively named: 3,5 diprenyl-4-hydroxycinnamic acid (artepillin C) (1), 3-prenyl-4-dihydroxycinnamoiloxy cinnamic acid (baccharin) (2), 3-prenyl-4-hydroxycinnamic acid (drupanin) (3), aromadendrin-4′-O-methyl-ether (4) and kaempferide (5).
The compounds were characterized by nuclear magnetic resonance and mass spectrometry. Their gastroprotective effects were evaluated against ethanol/HCl- and indomethacin-induced ulcer in mice. Further, histological, histochemical, oxidative and inflammatory parameters were analyzed at ulcerated tissue. Acid antisecretory activities also were also assessed.
Compound 2 did not reduce the ethanol/HCl- induced ulcer at 30 mg/kg (p.o), whereas the minimum oral gastroprotective doses of 1, 3, 4 and 5 were 0.3, 0.3, 3 and 3 mg/kg, respectively. Besides, these compounds prevented ethanol/HCl-induced ulcer by intraperitoneal route, as well as indomethacin-induced ulcer by oral route. The gastroprotection was accompanied by normalization of superoxide dismutase, catalase and glutathione-S-transferase activities and reduction in myeloperoxidase activity. Moreover, the compounds 4 and 5 increased the gastric mucin content and 1 reduced TNF amount. Furthermore, 1, 3, 4 and 5 decreased volume, pH, total acidity and pepsin activity of the gastric juice from rats.
Together, our findings showed a diversified mode of action elicited by 1, 3, 4 and 5 on the gastroprotection and contribute to explain the anti-ulcer activity reported for BGP.
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Endophytic fungi are associated with plant health and represent a remarkable source of potential of enzymes and bioactive compounds, but the diversity of endophytes remains uncertain and poorly ...explored, especially in Cactaceae, one of the most species‐rich families adapted to growing in arid and semi‐arid regions. The aim of this study was to conduct a systematic review on the diversity and bioprospecting of endophytic fungi from Cactaceae. We analysed peer‐reviewed articles from seven databases using PRISMA guidelines. The results showed that the Cactaceae family is a source of new taxa, but the diversity of endophytic fungi of Cactaceae is little explored, mainly the diversity among tissues and by metagenomics. Bioprospecting studies have shown that these microorganisms can be used in the production of enzymes and larvicidal and antifungal compounds. Our results are relevant as a starting point for researchers to develop studies that expand the knowledge of plant mycobiota in arid and semi‐arid ecosystems, as well as comprising a remarkable source of fungal compounds with several biotechnological applications.
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