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1.
Nat Methods ; 20(7): 1010-1020, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37202537

RESUMO

The Cell Tracking Challenge is an ongoing benchmarking initiative that has become a reference in cell segmentation and tracking algorithm development. Here, we present a significant number of improvements introduced in the challenge since our 2017 report. These include the creation of a new segmentation-only benchmark, the enrichment of the dataset repository with new datasets that increase its diversity and complexity, and the creation of a silver standard reference corpus based on the most competitive results, which will be of particular interest for data-hungry deep learning-based strategies. Furthermore, we present the up-to-date cell segmentation and tracking leaderboards, an in-depth analysis of the relationship between the performance of the state-of-the-art methods and the properties of the datasets and annotations, and two novel, insightful studies about the generalizability and the reusability of top-performing methods. These studies provide critical practical conclusions for both developers and users of traditional and machine learning-based cell segmentation and tracking algorithms.


Assuntos
Benchmarking , Rastreamento de Células , Rastreamento de Células/métodos , Aprendizado de Máquina , Algoritmos
2.
Artigo em Inglês | MEDLINE | ID: mdl-38189368

RESUMO

The oceans harbour a myriad of unknown micro-organisms that remain unstudied because of a failure to establish the right growth conditions under laboratory conditions. To overcome this limitation, an isolation effort inspired by the iChip was performed using marine sediments from Memória beach, Portugal. A novel strain, PMIC_1C1BT, was obtained and subjected to a polyphasic study. Cells of strain PMIC_1C1BT were Gram-positive, rod-shaped, divided by binary fission and formed colonies that were shiny light-yellow. Based on its full 16S rRNA gene sequence, strain PMIC_1C1BT was phylogenetically associated to the genus Microbacterium and its closest relatives were Microbacterium aurum KACC 15219T (98.55 %), Microbacterium diaminobutyricum RZ63T (98.48 %) and Microbacterium hatanonis JCM 14558T (98.13 %). Strain PMIC_1C1BT had a genome size of 2 761 607 bp with 67.71 mol% of G+C content and 2582 coding sequences, which is lower than the genus average. Strain PMIC_1C1BT grew from 15 to 30 °C, optimally at 25 °C, at pH 6.0 to 11.0, optimally between pH 6.0 and 8.0, and from 0 to 5 % (w/v) NaCl, optimally between 2.0 and 3.0 %. It grew with casamino acids, glutamine, methionine, N-acetylglucosamine, sodium nitrate, tryptophan, urea and valine as sole nitrogen sources, and arabinose and cellobiose as sole carbon sources. The major cellular fatty acids were anteiso-C15 : 0, iso-C16 : 0 and iso-C17 : 0. Genome mining revealed the presence of four biosynthetic gene clusters (BGCs) with low similarities to other known BCGs. Based on the polyphasic data, strain PMIC_1C1BT is proposed to represent a novel species, for which the name Microbacterium memoriense sp. nov. (=CECT 30366T=LMG 32350T) is proposed.


Assuntos
Actinomycetales , Microbacterium , Portugal , RNA Ribossômico 16S/genética , Composição de Bases , Ácidos Graxos/química , Filogenia , Análise de Sequência de DNA , DNA Bacteriano/genética , Técnicas de Tipagem Bacteriana , Bactérias
3.
Proc Natl Acad Sci U S A ; 118(1)2021 01 05.
Artigo em Inglês | MEDLINE | ID: mdl-33361149

RESUMO

Acetylcholine (ACh) promotes various cell migrations in vitro, but there are few investigations into this nonsynaptic role of ACh signaling in vivo. Here we investigate the function of a muscarinic receptor on an epithelial cell migration in Caenorhabditis elegans We show that the migratory gonad leader cell, the linker cell (LC), uses an M1/M3/M5-like muscarinic ACh receptor GAR-3 to receive extrasynaptic ACh signaling from cholinergic neurons for its migration. Either the loss of the GAR-3 receptor in the LC or the inhibition of ACh release from cholinergic neurons resulted in migratory path defects. The overactivation of the GAR-3 muscarinic receptor caused the LC to reverse its orientation through its downstream effectors Gαq/egl-30, PLCß/egl-8, and TRIO/unc-73 This reversal response only occurred in the fourth larval stage, which corresponds to the developmental time when the GAR-3::yellow fluorescent protein receptor in the membrane relocalizes from a uniform to an asymmetric distribution. These findings suggest a role for the GAR-3 muscarinic receptor in determining the direction of LC migration.


Assuntos
Acetilcolina/metabolismo , Movimento Celular/fisiologia , Receptores Muscarínicos/metabolismo , Acetilcolina/fisiologia , Animais , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/fisiologia , Células Epiteliais/metabolismo , Contração Muscular/fisiologia , Terminações Pré-Sinápticas/metabolismo , Receptores Muscarínicos/fisiologia , Transdução de Sinais
4.
Int J Mol Sci ; 25(8)2024 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-38673736

RESUMO

Abundant in citrus fruits, naringin (NAR) is a flavonoid that has a wide spectrum of beneficial health effects, including its anti-inflammatory activity. However, its use in the clinic is limited due to extensive phase I and II first-pass metabolism, which limits its bioavailability. Thus, lipid nanoparticles (LNPs) were used to protect and concentrate NAR in inflamed issues, to enhance its anti-inflammatory effects. To target LNPs to the CD44 receptor, overexpressed in activated macrophages, functionalization with hyaluronic acid (HA) was performed. The formulation with NAR and HA on the surface (NAR@NPsHA) has a size below 200 nm, a polydispersity around 0.245, a loading capacity of nearly 10%, and a zeta potential of about 10 mV. In vitro studies show the controlled release of NAR along the gastrointestinal tract, high cytocompatibility (L929 and THP-1 cell lines), and low hemolytic activity. It was also shown that the developed LNPs can regulate inflammatory mediators. In fact, NAR@NPsHA were able to decrease TNF-α and CCL-3 markers expression by 80 and 90% and manage to inhibit the effects of LPS by around 66% for IL-1ß and around 45% for IL-6. Overall, the developed LNPs may represent an efficient drug delivery system with an enhanced anti-inflammatory effect.


Assuntos
Anti-Inflamatórios , Flavanonas , Lipossomos , Nanopartículas , Flavanonas/farmacologia , Flavanonas/química , Humanos , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/química , Nanopartículas/química , Animais , Células THP-1 , Ácido Hialurônico/química , Ácido Hialurônico/farmacologia , Camundongos , Linhagem Celular , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Receptores de Hialuronatos/metabolismo , Composição de Medicamentos
5.
Traffic ; 22(9): 306-318, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34288289

RESUMO

The correct targeting and insertion of tail-anchored (TA) integral membrane proteins is critical for cellular homeostasis. TA proteins are defined by a hydrophobic transmembrane domain (TMD) at their C-terminus and are targeted to either the ER or mitochondria. Derived from experimental measurements of a few TA proteins, there has been little examination of the TMD features that determine localization. As a result, the localization of many TA proteins are misclassified by the simple heuristic of overall hydrophobicity. Because ER-directed TMDs favor arrangement of hydrophobic residues to one side, we sought to explore the role of geometric hydrophobic properties. By curating TA proteins with experimentally determined localizations and assessing hypotheses for recognition, we bioinformatically and experimentally verify that a hydrophobic face is the most accurate singular metric for separating ER and mitochondria-destined yeast TA proteins. A metric focusing on an 11 residue segment of the TMD performs well when classifying human TA proteins. The most inclusive predictor uses both hydrophobicity and C-terminal charge in tandem. This work provides context for previous observations and opens the door for more detailed mechanistic experiments to determine the molecular factors driving this recognition.


Assuntos
Retículo Endoplasmático , Eucariotos , Retículo Endoplasmático/metabolismo , Eucariotos/metabolismo , Humanos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Mitocôndrias/metabolismo , Transporte Proteico
6.
Artigo em Inglês | MEDLINE | ID: mdl-37079347

RESUMO

Organisms with distinctive biological features and cellular organization constitute the bacterial phylum Planctomycetota. In this study, we formally describe a novel isolate, strain ICT_H6.2T, isolated from sediment samples collected in the brackish environment of the Tagus River estuary (Portugal) using an iChip-based culturing technique. The 16S rRNA gene analysis placed this strain into the phylum Planctomycetota and family Lacipirellulaceae, with a similarity value of 98.0 % to its closest relative Aeoliella mucimassa Pan181T, the currently only known member of the genus. Strain ICT_H6.2T has a genome size of 7.8 Mbp and a DNA G+C content of 59.6 mol %. Strain ICT_H6.2T is heterotrophic, aerobic and capable of microaerobic growth. This strain grows from 10 to 37 °C and from pH 6.5 to 10.0, requires salt to grow, and can tolerate up to 4 % (w/v) NaCl. Diverse nitrogen and carbon sources are utilized for growth. Morphologically, strain ICT_H6.2T is white to beige pigmented, spherical to ovoid in shape and around 1.4×1.1 µm in size. The strain clusters mainly in aggregates and younger cells show motility. Ultrastructural studies showed a cell plan with cytoplasmatic membrane invaginations and unusual filamentous structures with hexagonal organization in transversal section. Morphological, physiological and genomic comparison between strain ICT_H6.2T and its closest relatives strongly suggests it represents a novel species within the genus Aeoliella, for which we propose the name Aeoliella straminimaris sp. nov., represented by strain ICT_H6.2T as the type strain (=CECT 30574T=DSM 114064T).


Assuntos
Ácidos Graxos , Planctomicetos , Ácidos Graxos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Composição de Bases , Filogenia , DNA Bacteriano/genética , Técnicas de Tipagem Bacteriana
7.
Artigo em Inglês | MEDLINE | ID: mdl-37486346

RESUMO

An isolation effort focused on sporogenous Actinomycetota from the Tagus estuary in Alcochete, Portugal, yielded a novel actinomycetal strain, designated MTZ3.1T, which was subjected to a polyphasic taxonomic study. MTZ3.1T is characterised by morphology typical of members of the genus Streptomyces, with light beige coloured substrate mycelium, which does not release pigments to the culture medium and with helicoidal aerial hyphae that differentiate into spores with a light-grey colour. The phylogeny of MTZ3.1T, based on the full 16S rRNA gene sequence, indicated that its closest relatives were Streptomyces alkaliterrae OF1T (98.48 %), Streptomyces chumphonensis KK1-2T (98.41 %), Streptomyces albofaciens JCM 4342T (98.34 %), Streoptomyces paromomycinus NBRC 15454T (98.34 %) and Streptomyces chrestomyceticus NRBC 13444T (98.34 %). Moreover, average nucleotide identity (ANI), average amino acid identity (AAI) and digital DNA-DNA hybridisation (dDDH) are below the species cutoff values (ANI 67.70 and 68.35 %, AAI 77.06 and 76.71 % and dDDH 22.10 and 21.50 % for S. alkaliterrae OF1T and S. chumphonensis KK1-2T, respectively). Whole genome sequencing revealed that MTZ3.1T has a genome of 5 644 485 bp with a DNA G+C content of 71.29 mol% and 5044 coding sequences. Physiologically, MTZ3.1T is strictly aerobic, able to grow at 15-37 °C, optimally at 25 °C and between pH5 and 8 and showed high salinity tolerance, growing with 0-10 %(w/v) NaCl. Major cellular fatty acids are C15 : 0, iso-C15 : 0, anteiso-C15 : 0 and iso-C16 : 0. Furthermore, it was able to utilise a variety of nitrogen and carbon sources. Antimicrobial screening indicated that MTZ3.1T has potent anti-Staphylococcus aureus activity. On the basis of the polyphasic data, MTZ3.1T is proposed to represent a novel species, Streptomyces meridianus sp. nov. (= CECT 30416T = DSM 114037T=LMG 32463T).


Assuntos
Ácidos Graxos , Streptomyces , Ácidos Graxos/química , RNA Ribossômico 16S/genética , Portugal , Estuários , Análise de Sequência de DNA , Filogenia , Composição de Bases , DNA Bacteriano/genética , Técnicas de Tipagem Bacteriana , Ácido Diaminopimélico/química , Águas Salinas , Fosfolipídeos/química
8.
Artigo em Inglês | MEDLINE | ID: mdl-37489568

RESUMO

A novel actinomycetal strain, designated M600PL45_2T, was isolated from marine sediments obtained from Ingleses beach, Porto, on the Northern Coast of Portugal and was subjected to a polyphasic taxonomic characterisation study. The here described Gram-reaction-positive strain is characterised by the production of a brown pigment in both solid and liquid medium and forms typical helical hyphae that differentiate into smooth spores. The results of a phylogenetic analysis based on the 16S rRNA gene sequence indicated that M600PL45_2T has a high similarity to two members of the genus Streptomyces, Streptomyces bathyalis ASO4wetT (98.51 %) and Streptomyces daqingensis NEAU ZJC8T (98.44 %). The genome of M600PL45_2T has a size of 6 695 159 bp, a DNA G+C content of 70.71 mol% and 5538 coding sequences. M600PL45_2T grows at 15-37 °C and with a maximal growth rate between 25 °C and 30 °C. Growth at pH 6.0 to 9.0 with the optimal range between 6.0 and 7.5 was observed. M600PL45_2T showed a high salinity tolerance, growing with 0-10 % (w/v) NaCl, with best growth with 1-3% (w/v) NaCl. Major cellular fatty acids are iso-C15:0 (25.03 %), anteiso-C15:0 (17.70) and iso-C16:0 (26.90 %). The novel isolate was able to grow in media containing a variety of nitrogen and carbon sources. An antimicrobial activity screening indicated that an extract of M600PL45_2T has inhibitory activity against Staphylococcus aureus. On the basis of the polyphasic data, M600PL45_2T (= CECT 30365T = DSM 114036T) is introduced as the type strain of a novel species, that we named Streptomyces marispadix sp. nov.


Assuntos
Ácidos Graxos , Cloreto de Sódio , Composição de Bases , Ácidos Graxos/química , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , DNA Bacteriano/genética , Técnicas de Tipagem Bacteriana , Sedimentos Geológicos
9.
Nat Rev Mol Cell Biol ; 12(4): 265-73, 2011 04.
Artigo em Inglês | MEDLINE | ID: mdl-21364682

RESUMO

The emerging field of computational morphodynamics aims to understand the changes that occur in space and time during development by combining three technical strategies: live imaging to observe development as it happens; image processing and analysis to extract quantitative information; and computational modelling to express and test time-dependent hypotheses. The strength of the field comes from the iterative and combined use of these techniques, which has provided important insights into plant development.


Assuntos
Processamento de Imagem Assistida por Computador/métodos , Imageamento Tridimensional/métodos , Microscopia Confocal/métodos , Desenvolvimento Vegetal , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Flores/genética , Flores/crescimento & desenvolvimento , Flores/metabolismo , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Cinética , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Meristema/genética , Meristema/crescimento & desenvolvimento , Meristema/metabolismo , Plantas/genética , Plantas/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Fatores de Tempo
10.
Antonie Van Leeuwenhoek ; 116(11): 1209-1225, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37737556

RESUMO

A bacterial strain was isolated from a brackish water sample of Tagus river, Alcochete, Portugal and was designated TO1_6T. It forms light pink colonies on M13 medium supplemented with N-acetylglucosamine. Cells are pear-shaped to spherical, form rosettes and divide by budding. Strain TO1_6T presents a mesophilic and neutrophilic profile, with optimum growth at 20 to 25 °C and pH 7.0 to 7.5, and vitamin supplementation is not required to promote its growth. The genome of the novel isolate is 7.77 Mbp in size and has a DNA G + C content of 56.3%. Based on its 16S rRNA gene sequence, this strain is affiliated with the phylum Planctomycetota. Further taxonomic characterization using additional phylogenetic markers, namely rpoB gene sequence (encoding the ß-subunit of the DNA-dependent RNA polymerase), as well as Percentage of conserved proteins, average nucleotide identity and average amino acid identity, suggest the affiliation of strain TO1_6T to the genus Stieleria, a recently described taxon in the family Pirellulaceae, order Pirellulales and class Planctomycetia. Based on the genotypic, phylogenetic and physiological characterization, we here describe a new species represented by the type strain TO1_6T (= CECT 30432T, = LMG 32465T), for which the name Stieleria tagensis sp. nov. is proposed.


Assuntos
Ácidos Graxos , Rios , Rios/microbiologia , Ácidos Graxos/análise , Fosfolipídeos/análise , Planctomicetos , Análise de Sequência de DNA , Filogenia , RNA Ribossômico 16S/genética , Portugal , DNA Bacteriano/genética , DNA Bacteriano/química , Técnicas de Tipagem Bacteriana
11.
Proc Natl Acad Sci U S A ; 117(44): 27400-27411, 2020 11 03.
Artigo em Inglês | MEDLINE | ID: mdl-33087579

RESUMO

Individual cell migration requires front-to-back polarity manifested by lamellipodial extension. At present, it remains debated whether and how membrane motility mediates this cell morphological change. To gain insights into these processes, we perform live imaging and molecular perturbation of migrating chick neural crest cells in vivo. Our results reveal an endocytic loop formed by circular membrane flow and anterograde movement of lipid vesicles, resulting in cell polarization and locomotion. Rather than clathrin-mediated endocytosis, macropinosomes encapsulate F-actin in the cell body, forming vesicles that translocate via microtubules to deliver actin to the anterior. In addition to previously proposed local conversion of actin monomers to polymers, we demonstrate a surprising role for shuttling of F-actin across cells for lamellipodial expansion. Thus, the membrane and cytoskeleton act in concert in distinct subcellular compartments to drive forward cell migration.


Assuntos
Actinas/metabolismo , Movimento Celular , Crista Neural/fisiologia , Pinocitose , Pseudópodes/metabolismo , Animais , Membrana Celular/metabolismo , Embrião de Galinha , Microscopia Intravital , Crista Neural/citologia , Imagem com Lapso de Tempo
12.
Artigo em Inglês | MEDLINE | ID: mdl-35687662

RESUMO

The phylum Planctomycetota is constituted by bacteria with unique features that are well adapted to a vast range of habitats. Here, we describe a novel planctomycete isolated from marine sediments collected on a beach in Matosinhos (Portugal) using an iChip-based culturing technique. Strain ICM_H10T forms beige-coloured colonies in modified M14 medium and its cells are spherical to ovoid in shape, stalked, rosette-forming and showing motility in a phase of the life cycle. Transmission electron microscopy observations showed a typical planctomycetal cell plan and cell division by budding. This strain requires salt for growth and grows in the range of 2.0-5.0 % (w/v) NaCl, from 20 to 37 °C, within a pH of 6.0-9.0 and is able to use diverse nitrogen and carbon sources. It is heterotrophic, aerobic and capable of microaerobic growth. This strain has a genome size of approximately 6.0 Mb and a G+C content of 58.1 mol%. A 16S rRNA gene-based phylogenetic analysis supports the association of strain ICM_H10T to the phylum Planctomycetota and the family Planctomycetaceae, as it shares only 96.8 and 96.4% similarity to its closest relatives Rubinisphaera italica Pan54T and Rubinisphaera brasiliensis IFAM 1448T, respectively. Other phylogenetic markers also support the separation of this strain into a novel species. Morphological, physiological and genomic comparisons between strain ICM_H10T and its closest relatives strongly suggest that ICM_H10T represents a new species of the genus Rubinisphaera, for which we propose the name Rubinisphaera margarita sp. nov., with ICM_H10T (=CECT 30326T=LMG 32234T) as type strain.


Assuntos
Ácidos Graxos , Sedimentos Geológicos , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Sedimentos Geológicos/microbiologia , Filogenia , Portugal , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
13.
Development ; 145(12)2018 06 26.
Artigo em Inglês | MEDLINE | ID: mdl-29945988

RESUMO

In situ hybridization based on the mechanism of the hybridization chain reaction (HCR) has addressed multi-decade challenges that impeded imaging of mRNA expression in diverse organisms, offering a unique combination of multiplexing, quantitation, sensitivity, resolution and versatility. Here, with third-generation in situ HCR, we augment these capabilities using probes and amplifiers that combine to provide automatic background suppression throughout the protocol, ensuring that reagents will not generate amplified background even if they bind non-specifically within the sample. Automatic background suppression dramatically enhances performance and robustness, combining the benefits of a higher signal-to-background ratio with the convenience of using unoptimized probe sets for new targets and organisms. In situ HCR v3.0 enables three multiplexed quantitative analysis modes: (1) qHCR imaging - analog mRNA relative quantitation with subcellular resolution in the anatomical context of whole-mount vertebrate embryos; (2) qHCR flow cytometry - analog mRNA relative quantitation for high-throughput expression profiling of mammalian and bacterial cells; and (3) dHCR imaging - digital mRNA absolute quantitation via single-molecule imaging in thick autofluorescent samples.


Assuntos
Hibridização In Situ/métodos , Animais , Embrião de Galinha , Escherichia coli/genética , Citometria de Fluxo , Perfilação da Expressão Gênica , Humanos , Imageamento Tridimensional , Sondas RNA/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reprodutibilidade dos Testes , Frações Subcelulares/metabolismo
14.
BMC Infect Dis ; 20(1): 114, 2020 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-32041537

RESUMO

BACKGROUND: Identifying patients with hepatitis C virus (HCV) infection and enhancing the cascade of care are essential for eliminating HCV infection. This study aimed to estimate the prevalence of positive anti-HCV serology in Brasilia, Brazil, and evaluate the efficiency of the cascade of care for HCV-positive individuals. METHODS: This cross-sectional study analyzed 57,697 rapid screening tests for hepatitis C in individuals aged > 40 years between June 2018 and June 2019. HCV-positive patients were contacted and scheduled to undergo the HCV RNA viral test, genotyping, and transient elastography. RESULTS: The prevalence of positive serology was 0.27%. Among 161 patients with positive anti-HCV serology, 124 (77%) were contacted, 109 (67.7%) were tested for HCV RNA viral load, and 69 (42.8%) had positive results. Genotype 1 (75%) was the most prevalent genotype. Among 65 patients (94.2%) who underwent transient elastography, 30 (46.2%) presented with advanced fibrosis. Additionally, of the 161 patients, 55 (34.1%) were referred for treatment, but only 39 (24.2%) complied, with 36 (22.4%) showing sustained virological response. By the end of the study, 16 patients were still awaiting to receive medication. CONCLUSIONS: The prevalence of HCV-positive patients was low in Brasilia, and the gaps in the cascade of care for these patients were significantly below the targets of HCV infection elimination. This study opens new avenues for eliminating HCV infection and suggests that partnerships with clinical laboratories to conduct anti-HCV tests are a useful strategy to improve HCV diagnosis. TRIAL REGISTRATION: Research Ethics Committee of the Faculty of Health Sciences of the University of Brasília - UNB (CAAE number 77818317.2.0000.0030) and by the Ethics Committee of the Health Science Teaching and Research Foundation - FEPECS/SES/DF (CAAE number 77818317.2.3001.5553).


Assuntos
Hepacivirus/isolamento & purificação , Hepatite C/diagnóstico , Hepatite C/epidemiologia , Programas de Rastreamento , Adulto , Idoso , Antivirais/uso terapêutico , Brasil/epidemiologia , Estudos Transversais , Feminino , Genótipo , Hepacivirus/efeitos dos fármacos , Hepacivirus/genética , Hepacivirus/imunologia , Hepatite C/tratamento farmacológico , Hepatite C/virologia , Humanos , Cirrose Hepática/epidemiologia , Cirrose Hepática/patologia , Masculino , Programas de Rastreamento/estatística & dados numéricos , Pessoa de Meia-Idade , Prevalência , Testes Sorológicos , Resposta Viral Sustentada
15.
Cell Tissue Res ; 377(3): 475-503, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31478138

RESUMO

The phylum Mollusca is one of the largest and more diversified among metazoan phyla, comprising many thousand species living in ocean, freshwater and terrestrial ecosystems. Mollusc-feeding biology is highly diverse, including omnivorous grazers, herbivores, carnivorous scavengers and predators, and even some parasitic species. Consequently, their digestive system presents many adaptive variations. The digestive tract starting in the mouth consists of the buccal cavity, oesophagus, stomach and intestine ending in the anus. Several types of glands are associated, namely, oral and salivary glands, oesophageal glands, digestive gland and, in some cases, anal glands. The digestive gland is the largest and more important for digestion and nutrient absorption. The digestive system of each of the eight extant molluscan classes is reviewed, highlighting the most recent data available on histological, ultrastructural and functional aspects of tissues and cells involved in nutrient absorption, intracellular and extracellular digestion, with emphasis on glandular tissues.


Assuntos
Sistema Digestório/ultraestrutura , Moluscos/ultraestrutura , Animais
16.
Int J Syst Evol Microbiol ; 68(4): 982-989, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29458463

RESUMO

An isolate, designated SPSPC-11T, with an optimum growth temperature of about 50 °C and an optimum pH for growth between 7.5 and 8.0, was recovered from a hot spring in central Portugal. Based on phylogenetic analysis of its 16S rRNA sequence, the new organism is most closely related to the species of the genus Thermonema but with a pairwise sequence similarity of <85 %. The isolate was orange-pigmented, formed non-motile long filaments and rod-shaped cells that stain Gram-negative. The organism was strictly aerobic, oxidase-positive and catalase-positive. The major fatty acids were iso-C15:0, iso-C15 : 0 2-OH and iso-C17 : 0 3-OH. The major polar lipids were one aminophospholipid, two aminolipids and three unidentified lipids. Menaquinone 7 was the major respiratory quinone. The DNA G+C content of strain SPSPC-11T was 37.6 mol% (draft genome sequence). The high quality draft genome sequence corroborated many of the phenotypic characteristics of strain SPSPC-11T. Based on genotypic, phylogenetic, physiological and biochemical characterization we describe a new species of a novel genus represented by strain SPSPC-11T (=CECT 9012T=LMG 29233T) for which we propose the name Raineya orbicola gen. nov., sp. nov. We also describe the family Raineyaceae to accommodate this new genus and species.


Assuntos
Bacteroidetes/classificação , Fontes Termais/microbiologia , Filogenia , Técnicas de Tipagem Bacteriana , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , Pigmentação , Portugal , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química
17.
Molecules ; 23(5)2018 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-29702573

RESUMO

Bioprospection of marine invertebrates has been predominantly biased by the biological richness of tropical regions, thus neglecting macro-organisms from temperate ecosystems. Species that were not the object of studies on their biochemical composition include the Heterobranchia gastropods Armina maculata, Armina tigrina and Aglaja tricolorata, inhabitants of the Portuguese Atlantic coastal waters. Here, we present for the first time the fatty acid profile of neutral lipids and homarine content of these three species. Qualitative and quantitative differences in the fatty acid content among species points to the existence of a fatty acid profile of neutral lipids, particularly of each genus. The results from cytotoxicity assays, using the acetonic extracts of the gastropods on human gastric adenocarcinoma (AGS) and human lung adenocarcinoma (A549) cell lines, revealed a pronounced cytotoxic effect of the A. tigrina extract on both cell lines (IC50 values of 68.75 and 69.77 µg mL−1 for AGS and A549, respectively). It is worth noting the significant reduction of NO levels in LPS-challenged RAW 264.7 macrophages exposed to A. tricolorata extract, at concentrations as low as 125 µg mL−1.


Assuntos
Anti-Inflamatórios/análise , Antineoplásicos/análise , Ácidos Graxos/análise , Gastrópodes/metabolismo , Macrófagos/efeitos dos fármacos , Ácidos Picolínicos/análise , Células A549 , Animais , Anti-Inflamatórios/metabolismo , Antineoplásicos/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Ácidos Graxos/metabolismo , Humanos , Lipopolissacarídeos/efeitos adversos , Macrófagos/citologia , Macrófagos/metabolismo , Camundongos , Óxido Nítrico/metabolismo , Ácidos Picolínicos/metabolismo , Células RAW 264.7
18.
Int J Syst Evol Microbiol ; 67(5): 1571-1576, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28036240

RESUMO

One strain of a novel genus and species of the order Planctomycetes, designated FC18T, was isolated from the epiphytic community of Fucusspiralis. This strain was non-pigmented in medium M13 but was slightly pink pigmented on medium M14, containing four-fold the levels of glucose, peptone and yeast extract of M13. The organism was primarily spherical, with unicellular non-motile forms and rosettes. The optimal temperature for growth was about 25 °C and the optimal pH was 7.5. FC18T was chemoorganotrophic and aerobic. Several sugars, polyols and amino acids were assimilated. The major fatty acids were C18 : 1ω9c, C14 : 0 and C16 : 0. The major polar lipids were phosphatidylglycerol (PG) and two unknown lipids. Menaquinone 5 (MK-5) was the main respiratory quinone, but MK-6 was also present. The results of the 16S rRNA gene sequence analysis confirmed the affiliation of this organism to the order Planctomycetales, family Planctomycetaceae, with Blastopirellula marina as the closest relative with only 86 % sequence similarity. On the basis of physiological, biochemical and chemotaxonomic characteristics we propose that FC18T(=LMG 29748T=DSM 26290T) represents a novel species of a novel genus of the family Planctomycetaceae for which we propose the name Mariniblastusfucicola gen. nov., sp. nov.


Assuntos
Filogenia , Planctomycetales/classificação , Alga Marinha/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfatidilgliceróis/química , Planctomycetales/genética , Planctomycetales/isolamento & purificação , Portugal , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/química
19.
Int J Syst Evol Microbiol ; 67(10): 3752-3758, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28895512

RESUMO

Strain EPL6T, a Gram-negative, motile, short rod was isolated from a propanil and 3,4-dichloroaniline enrichment culture produced from rice paddy soil. Based on the analyses of the 16S rRNA gene sequence, strain EPL6T was observed to be a member of the family Comamonadaceae, sharing the highest pairwise identity with type strains of the species Alicycliphilus denitrificans K601T (96.8 %) and Melaminivora alkalimesophila CY1T (96.8 %). Strain EPL6T was able to grow in a temperature range of 15-37 °C, pH 6-9 and in the presence of up to 4 % (w/v) NaCl and tested positive for catalase and oxidase reactions. The major respiratory quinone was Q8. The genomic DNA had a G+C content of 69.4±0.9 mol%. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol, and the major fatty acid methyl esters comprised C16 : 0, C18 : 1ω7c and summed feature 3 (C16 : 1ω7c/iso-C15 : 0 2-OH). Comparison of the genome sequence of strain EPL6T and of its closest neighbours, Melaminivora alkalimesophila CY1T and Alicycliphilus denitrificans K601T, yielded values of ANI ≤84.1 % and of AAI ≤80.3 %. Therefore, the genetic, phylogenetic, phenotypic and chemotaxonomic characteristics support the classification of this organism into a new taxon. Considering the genetic divergence of strain EPL6T from the type strains of the closest species, which belong to distinct genera, we propose a new genus within the family Comamonadaceae, named Oryzisolibacter propanilivorax gen. nov., sp. nov., represented by the isolate EPL6T as the type strain of the species (=LMG 28427T=CECT 8927T).


Assuntos
Filogenia , Propanil/metabolismo , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , Comamonadaceae/classificação , Comamonadaceae/genética , Comamonadaceae/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Oryza , Fosfolipídeos/química , Portugal , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/química
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