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1.
Mar Drugs ; 21(5)2023 May 19.
Artigo em Inglês | MEDLINE | ID: mdl-37233506

RESUMO

Industrial wastes with hazardous dyes serve as a major source of water pollution, which is considered to have an enormous impact on public health. In this study, an eco-friendly adsorbent, the porous siliceous frustules extracted from the diatom species Halamphora cf. salinicola, grown under laboratory conditions, has been identified. The porous architecture and negative surface charge under a pH of 7, provided by the various functional groups via Si-O, N-H, and O-H on these surfaces, revealed by SEM, the N2 adsorption/desorption isotherm, Zeta-potential measurement, and ATR-FTIR, respectively, made the frustules an efficient mean of removal of the diazo and basic dyes from the aqueous solutions, 74.9%, 94.02%, and 99.81% against Congo Red (CR), Crystal Violet (CV), and Malachite Green (MG), respectively. The maximum adsorption capacities were calculated from isotherms, as follows: 13.04 mg g-1, 41.97 mg g-1, and 33.19 mg g-1 against CR, CV, and MG, respectively. Kinetic and isotherm models showed a higher correlation to Pore diffusion and Sips models for CR, and Pseudo-Second Order and Freundlich models for CV and MG. Therefore, the cleaned frustules of the thermal spring-originated diatom strain Halamphora cf. salinicola could be used as a novel adsorbent of a biological origin against anionic and basic dyes.


Assuntos
Diatomáceas , Poluentes Químicos da Água , Corantes/química , Corantes de Rosanilina/química , Vermelho Congo , Água/química , Adsorção , Cinética , Concentração de Íons de Hidrogênio , Poluentes Químicos da Água/química
2.
Mar Drugs ; 20(4)2022 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-35447907

RESUMO

The marine pennate diatom Haslea ostrearia has long been known for its characteristic blue pigment marennine, which is responsible for the greening of invertebrate gills, a natural phenomenon of great importance for the oyster industry. For two centuries, this taxon was considered unique; however, the recent description of a new blue Haslea species revealed unsuspected biodiversity. Marennine-like pigments are natural blue dyes that display various biological activities-e.g., antibacterial, antioxidant and antiproliferative-with a great potential for applications in the food, feed, cosmetic and health industries. Regarding fundamental prospects, researchers use model organisms as standards to study cellular and physiological processes in other organisms, and there is a growing and crucial need for more, new and unconventional model organisms to better correspond to the diversity of the tree of life. The present work, thus, advocates for establishing H. ostrearia as a new model organism by presenting its pros and cons-i.e., the interesting aspects of this peculiar diatom (representative of benthic-epiphytic phytoplankton, with original behavior and chemodiversity, controlled sexual reproduction, fundamental and applied-oriented importance, reference genome, and transcriptome will soon be available); it will also present the difficulties encountered before this becomes a reality as it is for other diatom models (the genetics of the species in its infancy, the transformation feasibility to be explored, the routine methods needed to cryopreserve strains of interest).


Assuntos
Cosméticos , Diatomáceas , Ostreidae , Animais , Antioxidantes/farmacologia , Diatomáceas/fisiologia , Pigmentação
3.
Int J Mol Sci ; 23(19)2022 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-36233015

RESUMO

Nanostructures-assisted laser desorption/ionization mass spectrometry (NALDI-MS) is gaining attention for the analysis of a wide range of molecules. In this present investigation, Pseudostaurosira trainorii mediated biosynthesized iron-oxide nanoparticles (IONPs) have been utilized as nanostructures assisting ionization and desorption for laser desorption/ionization mass spectrometry (LDI-MS). The chain forming diatom, P. trainorii showed efficiency in the production of IONPs against 0.01 M Fe+3 (pH 2) aqueous solution at the intracellular and extracellular level. The whole biomass and external media turned dark orange in color after 3 days of reaction with Fe3+ solution. Scanning electron microscopic (SEM) images illustrated that the surface of Fe3+ exposed frustules of P. trainorii were entirely covered by synthesized nanostructures contrasting with the natural surface ornamentation of control cells. The IONPs loaded frustules also exhibited catalytic properties by decolorizing yellow colored nitrophenol after 3 h of reaction. Transmission electron microscopic (TEM) images confirmed that the produced particles are spindle-shaped with ~50-70 nm length and ~10-30 nm width. The biogenic IONPs were utilized as an inorganic matrix in LDI-MS and showed high sensitivity towards small molecules as glucose, alanine and triacylglycerols at nano- and picomolar level per spot, respectively. The presented biocompatible technique offers new perspectives in nanobiotechnology for the production of spindle-shaped IONPs that can be applied in future for the preparation of NALDI plates.


Assuntos
Diatomáceas , Nanopartículas , Alanina , Glucose , Ferro , Lasers , Nanopartículas/química , Nitrofenóis , Óxidos/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Triglicerídeos
4.
Biochem Biophys Res Commun ; 534: 261-265, 2021 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-33280817

RESUMO

3-methylglutaconic (3MGC) aciduria is associated with a growing number of discrete inborn errors of metabolism. Herein, an antibody-based approach to detection/quantitation of 3MGC acid has been pursued. When trans-3MGC acid conjugated keyhole limpet hemocyanin (KLH) was inoculated into rabbits a strong immune response was elicited. Western blot analysis provided evidence that immune serum, but not pre-immune serum, recognized 3MGC-conjugated bovine serum albumin (BSA). In competition ELISAs using isolated immune IgG, the limit of detection for free trans-3MGC acid was compared to that for cis-3MGC acid and four structurally related short-chain dicarboxylic acids. Surprisingly, cis-3MGC acid yielded a much lower limit of detection (∼0.1 mg/ml) than trans-3MGC acid (∼1.0 mg/ml) while all other dicarboxylic acids tested were poor competitors. The data suggest trans-3MGC- isomerized during, or after, conjugation to KLH such that the immunogen was actually comprised of KLH harboring a mixture of cis- and trans-3MGC haptens. To investigate this unexpected isomerization reaction, trans-3MGC CoA was prepared and incubated at 37 °C in the presence of BSA. Evidence was obtained that non-enzymatic isomerization of trans-3MGC CoA to cis-3MGC CoA precedes intramolecular catalysis to form cis-3MGC anhydride plus CoASH. Anhydride-dependent acylation of BSA generated 3MGCylated BSA, as detected by anti-3MGC immunoblot. The results presented provide an explanation for the unanticipated detection of 3MGCylated proteins in a murine model of primary 3MGC aciduria. Furthermore, non-enzymatic hydrolysis of cis-3MGC anhydride represents a potential source of cis-3MGC acid found in urine of subjects with 3MGC aciduria.


Assuntos
Glutaratos/química , Glutaratos/imunologia , Acilação , Animais , Coenzima A/metabolismo , Ácidos Dicarboxílicos/análise , Ácidos Dicarboxílicos/imunologia , Glutaratos/análise , Haptenos/imunologia , Hemocianinas/imunologia , Hemocianinas/metabolismo , Temperatura Alta , Soros Imunes/imunologia , Imunoglobulina G/imunologia , Isomerismo , Coelhos , Soroalbumina Bovina/imunologia
5.
Mol Phylogenet Evol ; 158: 106985, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33059066

RESUMO

The Bacillariaceae is a very species-rich family of raphid diatoms and includes the large and taxonomically difficult genus Nitzschia, whose species are often small-celled and finely structured and have few discrete morphological characters visible in the light microscope. The classification of Nitzschia is still mostly based on one developed in the second half of the 19th century by Grunow, who separated the genus into a series of sections largely on cell shape and symmetry, the position of the raphe, transverse extension of the fibulae, and folding of the valve. We assembled and analysed single-gene and concatenated alignments of nSSU, nLSU, rbcL, psbC and cox1 to test Grunow's and subsequent classifications and to examine selected morphological characters for their potential to help define monophyletic groups. The maximum likelihood trees were equivocal as to monophyly of the family itself but showed good support for each of eight main clades of Bacillariaceae, three of which corresponded more or less to existing genera (Hantzschia, Cylindrotheca and Bacillaria). The other five main clades and some subclades comprised groups of Nitzschia species or assemblies of Nitzschia species with other genera (Pseudo-nitzschia, Fragilariopsis, Neodenticula, Tryblionella, Psammodictyon). Relationships between most of the eight main clades were not resolved robustly but all analyses recovered Nitzschia as non-monophyletic. The Grunowian classification of Nitzschia into sections was not supported, though in some respects (e.g. treatment of sigmoid species) it is better than subsequent reclassifications. Several of the main clades and subclades are cryptic (lacking morphological synapomorphies) and homoplasy is common in both light microscopical and ultrastructural characters (to the extent that organisms initially assigned to the same species sometimes prove to belong to a different main clade). Nevertheless, some characters, including the structure of the raphe canal and girdle, seem to be sufficiently conservative evolutionarily to give a provisional estimate of relationships if molecular data are unavailable. No new formal classifications are proposed but various options are explored and research needs identified.


Assuntos
Diatomáceas/classificação , Cloroplastos/classificação , Cloroplastos/genética , Diatomáceas/genética , Diatomáceas/fisiologia , Complexo IV da Cadeia de Transporte de Elétrons/classificação , Complexo IV da Cadeia de Transporte de Elétrons/genética , Funções Verossimilhança , Microscopia Eletrônica de Varredura , Filogenia , RNA Ribossômico 18S/classificação , RNA Ribossômico 18S/genética , RNA Ribossômico 28S/classificação , RNA Ribossômico 28S/genética
6.
J Phycol ; 57(5): 1472-1491, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34165803

RESUMO

Presented here are new insights into the marine monoraphid diatom genera Schizostauron and Astartiella, based on molecular and morphological data, including descriptions of new species. Although no unambiguous morphological synapomorphies between the two genera are currently recognized, they are closely related by DNA sequence data. Heterovalvate frustules of Schizostauron are characterized by a bifid stauros on the raphe-bearing valve and intricate areolate occlusions on the sternum valve. In Astartiella, the raphe-bearing valve is characterized by a process resembling a fistula by morphology, while the sternum valve presents a particular striation pattern. Observations by light and electron microscopy were made, along with a molecular phylogenetic analysis using a three-gene (SSU, rbcL, and psbC) concatenated dataset. Three new Schizostauron species are described (S. kajotkei, S. rawaii, S. papilliareae), and two new combinations proposed (S. citronella and S. trachyderma) for species that were previously included either in Achnanthes and Cocconeis, respectively. Likewise, six new species of Astartiella (A. almalikii, A. bornmanii, A. chunlianlii, A. marksii, A. persica, and A. wangii) are described. Molecular results exclude Schizostauron and Astartiella from three clades of exclusively monoraphid diatoms, the Achnanthaceae, Cocconeidaceae, and Achnanthidiaceae, instead placing them in the Stauroneidaceae. Morphological features of Schizostauron and Astartiella, such as the stauros, fistula, and coaxial internal proximal raphe endings, are found in other genera in this clade, whereas the only common feature with monoraphid diatoms as whole group is the heterovalvy of frustules.


Assuntos
Diatomáceas , Fístula , Diatomáceas/genética , Microscopia Eletrônica , Filogenia
7.
Int J Mol Sci ; 22(13)2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34201641

RESUMO

A new catalyst based on biosilica doped with palladium(II) chloride nanoparticles was prepared and tested for efficient degradation of methyl orange (MO) in water solution under UV light excitation. The obtained photocatalyst was characterized by X-ray diffraction, TEM and N2 adsorption/desorption isotherms. The photocatalytic degradation process was studied as a function of pH of the solution, temperature, UV irradiation time, and MO initial concentration. The possibilities of recycling and durability of the prepared photocatalysts were also tested. Products of photocatalytic degradation were identified by liquid chromatography-mass spectrometry analyses. The photocatalyst exhibited excellent photodegradation activity toward MO degradation under UV light irradiation. Rapid photocatalytic degradation was found to take place within one minute with an efficiency of 85% reaching over 98% after 75 min. The proposed mechanism of photodegradation is based on the assumption that both HO• and O2•- radicals, as strongly oxidizing species that can participate in the dye degradation reaction, are generated by the attacks of photons emitted from diatom biosilica (photonic scattering effect) under the influence of UV light excitation. The degradation efficiency significantly increases as the intensity of photons emitted from biosilica is enhanced by palladium(II) chloride nanoparticles immobilized on biosilica (synergetic photonic scattering effect).


Assuntos
Compostos Azo/química , Diatomáceas/química , Paládio/química , Poluentes Químicos da Água/química , Catálise , Concentração de Íons de Hidrogênio , Cinética , Nanopartículas Metálicas/química , Microscopia Eletrônica de Transmissão , Fotólise , Reciclagem , Espectrometria de Massas por Ionização por Electrospray , Temperatura , Raios Ultravioleta , Difração de Raios X
8.
Int J Mol Sci ; 22(13)2021 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-34281209

RESUMO

We sequenced the plastid genomes of three diatoms from the genus Climaconeis, including two strains formerly designated as Climaconeis scalaris. At 208,097 and 216,580 bp, the plastid genomes of the latter strains are the largest ever sequenced among diatoms and their increased size is explained by the massive expansion of the inverted repeat region. Important rearrangements of gene order were identified among the two populations of Climaconeis cf. scalaris. The other sequenced Climaconeis chloroplast genome is 1.5 times smaller compared with those of the Climaconeis cf. scalaris strains and it features an usual quadripartite structure. The extensive structural changes reported here for the genus Climaconeis are compared with those previously observed for other algae and plants displaying large plastid genomes.


Assuntos
Expansão das Repetições de DNA , Diatomáceas/genética , Genomas de Plastídeos , Sequências Repetidas Invertidas , Diatomáceas/ultraestrutura
9.
Int J Mol Sci ; 22(20)2021 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-34681800

RESUMO

We provide for the first time the complete plastid and mitochondrial genomes of a monoraphid diatom: Schizostauron trachyderma. The mitogenome is 41,957 bp in size and displays two group II introns in the cox1 gene. The 187,029 bp plastid genome features the typical quadripartite architecture of diatom genomes. It contains a group II intron in the petB gene that overlaps the large single-copy and the inverted repeat region. There is also a group IB4 intron encoding a putative LAGLIDADG homing endonuclease in the rnl gene. The multigene phylogenies conducted provide more evidence of the proximity between S. trachyderma and fistula-bearing species of biraphid diatoms.


Assuntos
Diatomáceas/genética , Genoma Mitocondrial , Genomas de Plastídeos , Diatomáceas/classificação , Diatomáceas/citologia , Evolução Molecular , Fases de Leitura Aberta/genética , Filogenia , Análise de Sequência de DNA
10.
J Biol Chem ; 294(10): 3634-3646, 2019 03 08.
Artigo em Inglês | MEDLINE | ID: mdl-30635405

RESUMO

Amyloid deposits of apolipoprotein A-I (apoA-I) and inflammation are common in atherosclerotic arteries. In this study, we investigated the interplay between oxidation of apoA-I methionine residues (Met(O)-ApoA-I), a known amyloidogenic modification of apoA-I, and the inflammatory response of immune cells. Soluble pre-fibrillar Met(O)-ApoA-I, but not apoA-I, induced intracellular accumulation of pro-interleukin (IL)-1ß and secretion of the pro-inflammatory cytokines tumor necrosis factor α (TNFα) and IL-6 in mouse bone marrow-derived macrophages (BMDMs) and human primary monocytes. Additionally, secretion of mature IL-1ß was also activated in human monocytes. The pro-inflammatory activity of Met(O)-ApoA-I was Toll-like receptor 4 (TLR4)-dependent and CD36-independent and was solely determined by oxidation of apoA-I methionine residues, in particular Met-86 and Met-148. In contrast, amyloid fibrils or reconstituted high-density lipoproteins (HDLs) generated from Met(O)-ApoA-I did not induce cytokine production in BMDMs. Although lipid-free Met(O)-ApoA-I remained functional in extracting lipids from cells and generating HDL, it gained strong pro-inflammatory properties that may aggravate local inflammation in the arteries and atherosclerosis. Our study indicates that oxidation of apoA-I methionine residues produces a potent danger-associated molecular pattern capable of stimulating pro-inflammatory cytokine secretion at levels similar to those induced by known pathogen-associated molecular patterns, such as lipopolysaccharide.


Assuntos
Apolipoproteína A-I/química , Apolipoproteína A-I/metabolismo , Metionina/metabolismo , Animais , Humanos , Inflamação/metabolismo , Camundongos , Oxirredução , Receptor 4 Toll-Like/metabolismo
11.
Biochem Biophys Res Commun ; 533(3): 573-579, 2020 12 10.
Artigo em Inglês | MEDLINE | ID: mdl-32981676

RESUMO

Intricate mesoporous biosilica has many biomedical applications as a nanocarrier. However, its potential use in photodynamic therapy (PDT) has received little attention. This work reports the first fabrication of bio-engineered materials by covalently conjugating C-phycocyanin (C-PC), a natural photosensitizer, to biosilica for the PDT of tumor-associated macrophages. The resulting hybrid material showed outstanding photodynamic activity under 620 nm laser irradiation. Furthermore, it enhanced the relatively weak photodynamic effect of C-PC. This study also explored methods of biofunctionalizing biosilica for cancer phototherapy, a new pharmacological application of non-toxic C-PC.


Assuntos
Neoplasias/tratamento farmacológico , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Ficocianina/química , Dióxido de Silício/química , Animais , Sobrevivência Celular , Camundongos , Células RAW 264.7
12.
Mol Phylogenet Evol ; 148: 106808, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32243996

RESUMO

Although previous phylogenetic analyses suggested that the araphid diatom family Plagiogrammaceae is monophyletic, there is still not a clear understanding of relationships among the genera, and the taxonomy of several genera--Dimeregramma and Plagiogramma--remains questionable in light of paraphyly for both genera using molecular and morphological data. We have expanded the available DNA for molecular work for dozens of plagiogrammacean clones and analyzed 29 morphological characters from plagiogrammarian taxa and closely related genera, to increase understanding of the evolutionary history and systematics of the family and re-evaluate the current taxonomical classification of plagiogrammacean genera. The addition of more taxa and more data confirm the results from previous molecular phylogenies: most plagiogrammacean genera are monophyletic, except for Dimeregramma and Plagiogramma. Interestingly, the morphological analysis resolves only Talaroneis and Glyphodesmis as monophyletic. Given these results, we feel there is limited support for retaining Dimeregramma and Plagiogramma as distinct genera, and formally propose amending Plagiogramma and transferring six Dimeregramma species. As the Plagiogrammaceae is also one of the first-diverging clades of pennate diatoms, we also used these molecular data to estimate the age of the family, based on multiple calibration points derived from fossil taxa within or close to the Plagiogrammaceae. The results indicated that the Plagiogrammaceae evolved more than 114 million year ago and its diversification appears to correspond to a time of climate cooling. Additionally, we described a new monotypic genus (Coccinelloidea) with one new species C. gracilis, and five new species within established genera, e.g. Plagiogramma marginalis, Plagiogramma harenae, Plagiogramma porcipellis, Neofragilaria montgomeryii and Psammogramma anacarae.


Assuntos
Diatomáceas/classificação , Diatomáceas/genética , Filogenia , Animais , Teorema de Bayes , Mudança Climática , Diatomáceas/citologia , Diatomáceas/ultraestrutura , Fósseis , Análise de Sequência de DNA
13.
J Phycol ; 56(4): 953-978, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32259285

RESUMO

Detailed morphological documentation is provided for established Proschkinia taxa, including the generitype, P. bulnheimii, and P. complanata, P. complanatula, P. complanatoides and P. hyalosirella, and six new species. All established taxa are characterized from original material from historical collections. The new species described in this paper (P. luticola, P. staurospeciosa, P. impar, P. modesta, P. fistulispectabilis, and P. rosowskii) were isolated from the Western Pacific (Yellow Sea coast of Korea) and the Atlantic (Scottish and Texas coasts). Thorough documentation of the frustule, valve and protoplast architecture revealed the combination of characters diagnostic of the genus Proschkinia: a single-lobed chloroplast; a broad girdle composed of U-shaped, perforated bands; the position of the conopeate raphe-sternum relative to the external and internal valve surface; and the presence of an occluded process through the valve, termed the "fistula". Seven strains of Proschkinia were grown in culture and five of these were sequenced for nuclear ribosomal SSU and plastid-encoded rbcL. Phylogenetic analysis recovered a clade of Proschkinia with Fistulifera, another fistula-bearing diatom genus, and together these were sister to a clade formed of the Stauroneidaceae; in turn, all of these were sister to a clade composed of Parlibellus and two monoraphid genera Astartiella and Schizostauron. Despite morphological similarities between Proschkinia and the Naviculaceae, these two taxa are distant in our analysis. We document the variation in the morphology of Proschkinia, including significant variability in the fistula, suggesting that fistula ultrastructure might be one of the key features for species identification within the genus.


Assuntos
Diatomáceas , Filogenia , República da Coreia
14.
Invest New Drugs ; 37(6): 1309, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31032525

RESUMO

The authors would like to note an omission of disclosure in this paper. Author JDS is cofounder, equity-holder, and consultant of GILTRx Therapeutics.

15.
FASEB J ; 32(6): 3149-3165, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29401604

RESUMO

Apolipoprotein A-I (apoA-I) shares with other exchangeable apolipoproteins a high level of structural plasticity. In the lipid-free state, the apolipoprotein amphipathic α-helices interact intra- and intermolecularly, providing structural stabilization by self-association. We have reported that lipid-free apoA-I becomes amyloidogenic upon physiologically relevant (myeloperoxidase-mediated) Met oxidation. In this study, we established that Met oxidation promotes amyloidogenesis by reducing the stability of apoA-I monomers and irreversibly disrupting self-association. The oxidized apoA-I monomers also exhibited increased cellular cholesterol release capacity and stronger association with macrophages, compared to nonoxidized apoA-I. Of physiologic relevance, preformed oxidized apoA-I amyloid fibrils induced amyloid formation in nonoxidized apoA-I. This process was enhanced when self-association of nonoxidized apoA-I was disrupted by thermal treatment. Solid state NMR analysis revealed that aggregates formed by seeded nonoxidized apoA-I were structurally similar to those formed by the oxidized protein, featuring a ß-structure-rich amyloid fold alongside α-helices retained from the native state. In atherosclerotic lesions, the conditions that promote apoA-I amyloid formation are readily available: myeloperoxidase, active oxygen species, low pH, and high concentration of lipid-free apoA-I. Our results suggest that even partial Met oxidation of apoA-I can nucleate amyloidogenesis, thus sequestering and inactivating otherwise antiatherogenic and HDL-forming apoA-I.-Witkowski, A., Chan, G. K. L., Boatz, J. C., Li, N. J., Inoue, A. P., Wong, J. C., van der Wel, P. C. A., Cavigiolio, G. Methionine oxidized apolipoprotein A-I at the crossroads of HDL biogenesis and amyloid formation.


Assuntos
Amiloide/química , Apolipoproteína A-I/química , Lipoproteínas HDL/química , Metionina/química , Amiloide/metabolismo , Apolipoproteína A-I/metabolismo , Aterosclerose/metabolismo , Aterosclerose/patologia , Humanos , Lipoproteínas HDL/metabolismo , Metionina/metabolismo , Ressonância Magnética Nuclear Biomolecular , Oxirredução , Peroxidase/química , Peroxidase/metabolismo
16.
J Phycol ; 55(2): 425-441, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30615190

RESUMO

New molecular and morphological insights are presented on Campylosira africana and two new species, Extubocellulus cupola sp. nov. and Plagiogrammopsis castigatus sp. nov. Species descriptions were based on LM/SEM micrographs and a data set with concatenated sequences of SSU, rbcL and psbC loci constructed from 56 cymatosiroid strains isolated from global geographic locations. Extubocellulus cupola is distinguished by a mesh-like marginal ridge and dome-shaped areolation, and P. castigatus, by a prominent marginal extension at the valve center. Campylosira africana, a species from South Africa described by Professor Malcolm Giffen, has never been documented through LM or SEM, which led some authors to place it in synonymy under Campylosira cymbelliformis. We confirm that C. africana is a distinct species and provide microphotographic documentation and DNA sequences. We present morphological evidence for the well-known cymatosiroid species Extubocellulus spinifer possessing complete pili, and we emend the generic description of Extubocellulus.


Assuntos
Diatomáceas , Filogenia , África do Sul
17.
Invest New Drugs ; 36(5): 743-754, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29335887

RESUMO

Neuroblastoma is a childhood malignancy that accounts for approximately 15% of childhood cancer deaths. Only 20-35% of children with metastatic neuroblastoma survive with standard therapy. Identification of more effective therapies is essential to improving the outcome of children with high-stage disease. Sphingadienes (SD) are growth-inhibitory sphingolipids found in natural sources including soy. They exhibit chemopreventive activity in mouse models of colon cancer, where they mediate cytotoxicity by inhibiting key pro-carcinogenic signaling pathways. In this study, the effect of SD on neuroblastoma was analyzed. Low micromolar concentrations of SD were cytotoxic to transformed and primary neuroblastoma cells independently of N-Myc amplification status. SD induced both caspase-dependent apoptosis and autophagy in neuroblastoma cells. However, only inhibition of caspase-dependent apoptosis protected neuroblastoma cells from SD-mediated cytotoxicity. SD also inhibited AKT activation in neuroblastoma cells as shown by reduced phosphorylated AKT levels. Pre-treatment with insulin attenuated SD-mediated cytotoxicity in vitro. SD-loaded nanoparticles (NP) administered parenterally to immunodeficient mice carrying neuroblastoma xenografts resulted in cytotoxic levels of SD in the circulation and significantly reduced tumor growth compared to vehicle-treated controls. Analysis of tumor extracts demonstrated reduced AKT activation in tumors of mice treated with SD-NP compared to controls treated with empty NP. Our findings indicate SD are novel potential chemotherapeutic agents that promote neuroblastoma cell death and reduce tumorigenicity in vivo.


Assuntos
Antineoplásicos/administração & dosagem , Nanopartículas/administração & dosagem , Neuroblastoma/tratamento farmacológico , Esfingolipídeos/administração & dosagem , Animais , Antineoplásicos/sangue , Antineoplásicos/farmacocinética , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Feminino , Humanos , Camundongos SCID , Neuroblastoma/metabolismo , Neuroblastoma/patologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Esfingolipídeos/sangue , Esfingolipídeos/farmacocinética , Carga Tumoral/efeitos dos fármacos
18.
Arch Biochem Biophys ; 630: 38-46, 2017 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-28754322

RESUMO

Wnt signaling is essential for embryonic development and adult homeostasis in multicellular organisms. A conserved feature among Wnt family proteins is the presence of two structural domains. Within the N-terminal (NT) domain there exists a motif that is superimposable upon saposin-like protein (SAPLIP) family members. SAPLIPs are found in plants, microbes and animals and possess lipid surface seeking activity. To investigate the function of the Wnt3a saposin-like subdomain (SLD), recombinant SLD was studied in isolation. Bacterial expression of this Wnt fragment was achieved only when the core SLD included 82 NT residues of Wnt3a (NT-SLD). Unlike SAPLIPs, NT-SLD required the presence of detergent to achieve solubility at neutral pH. Deletion of two hairpin loop extensions present in NT-SLD, but not other SAPLIPs, had no effect on the solubility properties of NT-SLD. Far UV circular dichroism spectroscopy of NT-SLD yielded 50-60% α-helix secondary structure. Limited proteolysis of isolated NT-SLD in buffer and detergent micelles showed no differences in cleavage kinetics. Unlike prototypical saposins, NT-SLD exhibited weak membrane-binding affinity and lacked cell lytic activity. In cell-based canonical Wnt signaling assays, NT-SLD was unable to induce stabilization of ß-catenin or modulate the extent of ß-catenin stabilization induced by full-length Wnt3a. Taken together, the results indicate neighboring structural elements within full-length Wnt3a affect SLD conformational stability. Moreover, SLD function(s) in Wnt proteins appear to have evolved away from those commonly attributed to SAPLIP family members.


Assuntos
Proteína Wnt3A/química , Humanos , Lipídeos de Membrana/genética , Lipídeos de Membrana/metabolismo , Ligação Proteica , Domínios Proteicos , Estrutura Secundária de Proteína , Proteína Wnt3A/genética , Proteína Wnt3A/metabolismo
19.
J Phycol ; 53(2): 342-360, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-27988926

RESUMO

The family Cymatosiraceae, composed of two subfamilies (Cymatosiroideae and Extubocelluloideae), are elongate, non-pennate diatoms and are commonly reported in marine planktonic and benthic assemblages all over the world. The combination of the gross bilateral symmetry of the frustule and radial symmetry of the valve poration, distinct pore fields at the valve apices and DNA sequence data place this family in the controversial diatom class Mediophyceae, sister to the pennate diatoms. The relationships of the Cymatosiraceae within the Mediophyceae, and the monophyly of the two cymatosiroid subfamilies, have become less stable with the addition of new DNA sequence data. In this paper, we examined 24 cymatosiraceaen strains, including the new taxa Lambertocellus africana (Dabek & Witkowski) Dabek, Witkowski & Ashworth comb. nov., Leyanella probus Ashworth, Dabek & Witkowski sp. nov., and Leyanella pauciporis Ashworth, Dabek & Park sp. nov. to create the most complete molecular phylogeny of the Cymatosiraceae to date, using a three-gene (SSU, rbcL and psbC) data set. The results of the phylogenetic analysis supported the monophyly of the Cymatosiraceae, but not the monophyly of the subfamilies Cymatosiroideae and Extubocelluloideae. The phylogenetic analysis also suggested a close relationship of the Cymatosiraceae to the family Eupodiscaceae. The phylogenetic results have lead us to re-evaluate the taxonomy of L. africana, which is sister to Cymatosira lorenziana rather than Minutocellus as suggested in the original description.


Assuntos
Diatomáceas/genética , Evolução Molecular , Análise de Sequência de DNA/métodos , Diatomáceas/classificação , Família Multigênica/genética , Filogenia
20.
J Biol Chem ; 290(17): 10958-71, 2015 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-25759391

RESUMO

High plasma levels of apolipoprotein A-I (apoA-I) correlate with cardiovascular health, whereas dysfunctional apoA-I is a cause of atherosclerosis. In the atherosclerotic plaques, amyloid deposition increases with aging. Notably, apoA-I is the main component of these amyloids. Recent studies identified high levels of oxidized lipid-free apoA-I in atherosclerotic plaques. Likely, myeloperoxidase (MPO) secreted by activated macrophages in atherosclerotic lesions is the promoter of such apoA-I oxidation. We hypothesized that apoA-I oxidation by MPO levels similar to those present in the artery walls in atherosclerosis can promote apoA-I structural changes and amyloid fibril formation. ApoA-I was exposed to exhaustive chemical (H2O2) oxidation or physiological levels of enzymatic (MPO) oxidation and incubated at 37 °C and pH 6.0 to induce fibril formation. Both chemically and enzymatically oxidized apoA-I produced fibrillar amyloids after a few hours of incubation. The amyloid fibrils were composed of full-length apoA-I with differential oxidation of the three methionines. Met to Leu apoA-I variants were used to establish the predominant role of oxidation of Met-86 and Met-148 in the fibril formation process. Importantly, a small amount of preformed apoA-I fibrils was able to seed amyloid formation in oxidized apoA-I at pH 7.0. In contrast to hereditary amyloidosis, wherein specific mutations of apoA-I cause protein destabilization and amyloid deposition, oxidative conditions similar to those promoted by local inflammation in atherosclerosis are sufficient to transform full-length wild-type apoA-I into an amyloidogenic protein. Thus, MPO-mediated oxidation may be implicated in the mechanism that leads to amyloid deposition in the atherosclerotic plaques in vivo.


Assuntos
Amiloide/metabolismo , Apolipoproteína A-I/metabolismo , Aterosclerose/metabolismo , Peroxidase/metabolismo , Placa Aterosclerótica/metabolismo , Amiloide/genética , Apolipoproteína A-I/genética , Aterosclerose/genética , Aterosclerose/patologia , Humanos , Peróxido de Hidrogênio/farmacologia , Concentração de Íons de Hidrogênio , Metionina/genética , Metionina/metabolismo , Oxidantes/farmacologia , Oxirredução/efeitos dos fármacos , Peroxidase/genética , Placa Aterosclerótica/genética , Placa Aterosclerótica/patologia
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