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1.
Plant Cell Environ ; 47(9): 3638-3653, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38757412

RESUMO

Salinity tolerance requires coordinated responses encompassing salt exclusion in roots and tissue/cellular compartmentation of salt in leaves. We investigated the possible control points for salt ions transport in roots and tissue tolerance to Na+ and Cl- in leaves of two contrasting mungbean genotypes, salt-tolerant Jade AU and salt-sensitive BARI Mung-6, grown in nonsaline and saline (75 mM NaCl) soil. Cryo-SEM X-ray microanalysis was used to determine concentrations of Na, Cl, K, Ca, Mg, P, and S in various cell types in roots related to the development of apoplastic barriers, and in leaves related to photosynthetic performance. Jade AU exhibited superior salt exclusion by accumulating higher [Na] in the inner cortex, endodermis, and pericycle with reduced [Na] in xylem vessels and accumulating [Cl] in cortical cell vacuoles compared to BARI Mung-6. Jade AU maintained higher [K] in root cells than BARI Mung-6. In leaves, Jade AU maintained lower [Na] and [Cl] in chloroplasts and preferentially accumulated [K] in mesophyll cells than BARI Mung-6, resulting in higher photosynthetic efficiency. Salinity tolerance in Jade AU was associated with shoot Na and Cl exclusion, effective regulation of Na and Cl accumulation in chloroplasts, and maintenance of high K in root and leaf mesophyll cells.


Assuntos
Cloretos , Cloroplastos , Células do Mesofilo , Folhas de Planta , Raízes de Plantas , Potássio , Tolerância ao Sal , Sódio , Vigna , Raízes de Plantas/metabolismo , Raízes de Plantas/fisiologia , Cloroplastos/metabolismo , Sódio/metabolismo , Folhas de Planta/metabolismo , Folhas de Planta/fisiologia , Células do Mesofilo/metabolismo , Potássio/metabolismo , Cloretos/metabolismo , Vigna/metabolismo , Vigna/fisiologia , Fotossíntese , Transporte Biológico
2.
Environ Sci Technol ; 2024 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-38340051

RESUMO

Here, isotopically labeled 68ZnO NPs (ZnO NPs) and 68ZnO NPs with a thin 68Zn3(PO4)2 shell (ZnO_Ph NPs) were foliarly applied (40 µg Zn) to pepper plants (Capsicum annuum) to determine the effect of surface chemistry of ZnO NPs on the Zn uptake and systemic translocation to plant organs over 6 weeks. Despite similar dissolution of both Zn-based NPs after 3 weeks, the Zn3(PO4)2 shell on ZnO_Ph NPs (48 ± 12 nm; -18.1 ± 0.6 mV) enabled a leaf uptake of 2.31 ± 0.34 µg of Zn, which is 2.7 times higher than the 0.86 ± 0.18 µg of Zn observed for ZnO NPs (26 ± 8 nm; 14.6 ± 0.4 mV). Further, ZnO_Ph NPs led to higher Zn mobility and phloem loading, while Zn from ZnO NPs was stored in the epidermal tissues, possibly through cell wall immobilization as a storage strategy. These differences led to higher translocation of Zn from the ZnO_Ph NPs within all plant compartments. ZnO_Ph NPs were also more persistent as NPs in the exposed leaf and in the plant stem over time. As a result, the treatment of ZnO_Ph NPs induced significantly higher Zn transport to the fruit than ZnO NPs. As determined by spICP-TOFMS, Zn in the fruit was not in the NP form. These results suggest that the Zn3(PO4)2 shell on ZnO NPs can help promote the transport of Zn to pepper fruits when foliarly applied. This work provides insight into the role of Zn3(PO4)2 on the surface of ZnO NPs in foliar uptake and in planta biodistribution for improving Zn delivery to edible plant parts and ultimately improving the Zn content in food for human consumption.

3.
Neurochem Res ; 48(8): 2406-2423, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-36976393

RESUMO

The available data on the localization of transforming growth factor beta1 (TGF-ß1), glial cell line-derived neurotrophic factor (GDNF), and platelet-derived growth factor-BB (PDGF-BB) in the adult primate and human central nervous system (CNS) are limited and lack comprehensive and systematic information. This study aimed to investigate the cellular localization and distribution of TGF-ß1, GDNF, and PDGF-BB in the CNS of adult rhesus macaque (Macaca mulatta). Seven adult rhesus macaques were included in the study. The protein levels of TGF-ß1, PDGF-BB, and GDNF in the cerebral cortex, cerebellum, hippocampus, and spinal cord were analyzed by western blotting. The expression and location of TGF-ß1, PDGF-BB, and GDNF in the brain and spinal cord was examined by immunohistochemistry and immunofluorescence staining, respectively. The mRNA expression of TGF-ß1, PDGF-BB, and GDNF was detected by in situ hybridization. The molecular weight of TGF-ß1, PDGF-BB, and GDNF in the homogenate of spinal cord was 25 KDa, 30 KDa, and 34 KDa, respectively. Immunolabeling revealed GDNF was ubiquitously distributed in the cerebral cortex, hippocampal formation, basal nuclei, thalamus, hypothalamus, brainstem, cerebellum, and spinal cord. TGF-ß1 was least distributed and found only in the medulla oblongata and spinal cord, and PDGF-BB expression was also limited and present only in the brainstem and spinal cord. Besides, TGF-ß1, PDGF-BB, and GDNF were localized in the astrocytes and microglia of spinal cord and hippocampus, and their expression was mainly found in the cytoplasm and primary dendrites. The mRNA of TGF-ß1, PDGF-BB, and GDNF was localized to neuronal subpopulations in the spinal cord and cerebellum. These findings suggest that TGF-ß1, GDNF and PDGF-BB may be associated with neuronal survival, neural regeneration and functional recovery in the CNS of adult rhesus macaques, providing the potential insights into the development or refinement of therapies based on these factors.


Assuntos
Fator Neurotrófico Derivado de Linhagem de Célula Glial , Fator de Crescimento Transformador beta1 , Animais , Becaplermina , Macaca mulatta/metabolismo , RNA Mensageiro , Medula Espinal/metabolismo , Fator de Crescimento Transformador beta1/metabolismo
4.
BMC Neurosci ; 23(1): 20, 2022 03 30.
Artigo em Inglês | MEDLINE | ID: mdl-35354428

RESUMO

BACKGROUND: C-C motif chemokine ligand 2 (CCL2) is reported to be involved in the pathogenesis of various neurological and/or psychiatric diseases. Tissue or cellular expression of CCL2, in normal or pathological condition, may play an essential role in recruiting monocytes or macrophages into targeted organs, and be involved in a certain pathogenic mechanism. However, few studies focused on tissue and cellular distribution of the CCL2 peptide in brain grey and white matters (GM, WM), and the changes of the GM and WM cellular CCL2 level in septic or endotoxic encephalopathy was not explored. Hence, the CCL2 cellular distribution in the front brain cortex and the corpus callosum (CC) was investigated in the present work by using immunofluorescent staining. RESULTS: (1) CCL2 like immunoreactivity (CCL2-ir) in the CC is evidently higher than the cortex. When the measurement includes ependymal layer attached to the CC, CCL2-ir intensity is significantly higher than cortex. (2) Structures in perivascular areas, most of them are GFAP positive, contribute major CCL2-ir positive profiles in both GM and WM, but apparently more in the CC, where they are bilaterally distributed in the lateral CC between the cingulate cortex and ventricles. (3) The neuron-like CCL2-ir positive cells in cortex are significantly more than in the CC, and that number is significantly increased in the cortex following systemic lipopolysaccharide (LPS), but not in the CC. (4) In addition to CCL2-ir positive perivascular rings, more CCL2-ir filled cashew shape elements are observed, probably inside of microvasculature, especially in the CC following systemic LPS. (5) Few macrophage/microglia marker-Iba-1 and CCL2-ir co-labeled structures especially the soma is found in normal cortex and CC; the co-localizations are significantly augmented following systemic LPS, and co-labeled amoeba like somata are presented. (6) CCL2-ir and astrocyte marker GFAP or Iba-1 double labeled structures are also observed within the ependymal layer. No accumulation of neutrophils was detected. CONCLUSION: There exist differences in the cellular distribution of the CCL2 peptide in frontal cortex GM and subcortical WM-CC, in both the physiological condition and experimental endotoxemia. Which might cause different pathological change in the GM and WM.


Assuntos
Corpo Caloso , Lipopolissacarídeos , Animais , Lobo Frontal , Ligantes , Microglia/metabolismo
5.
Plant Cell Environ ; 45(5): 1490-1506, 2022 05.
Artigo em Inglês | MEDLINE | ID: mdl-35128687

RESUMO

Halophytes accumulate and sequester high concentrations of salt in vacuoles while maintaining lower levels of salt in the cytoplasm. The current data on cellular and subcellular partitioning of salt in halophytes are, however, limited to only a few dicotyledonous C3 species. Using cryo-scanning electron microscopy X-ray microanalysis, we assessed the concentrations of Na, Cl, K, Ca, Mg, P and S in various cell types within the leaf-blades of a monocotyledonous C4 halophyte, Rhodes grass (Chloris gayana). We also linked, for the first time, elemental concentrations in chloroplasts of mesophyll and bundle sheath cells to their ultrastructure and photosynthetic performance of plants grown in nonsaline and saline (200 mM NaCl) conditions. Na and Cl accumulated to the highest levels in xylem parenchyma and epidermal cells, but were maintained at lower concentrations in photosynthetically active mesophyll and bundle sheath cells. Concentrations of Na and Cl in chloroplasts of mesophyll and bundle sheath cells were lower than in their respective vacuoles. No ultrastructural changes were observed in either mesophyll or bundle sheath chloroplasts, and photosynthetic activity was maintained in saline conditions. Salinity tolerance in Rhodes grass is related to specific cellular Na and Cl distributions in leaf tissues, and the ability to regulate Na and Cl concentrations in chloroplasts.


Assuntos
Tolerância ao Sal , Plantas Tolerantes a Sal , Cloroplastos/metabolismo , Íons/metabolismo , Folhas de Planta/metabolismo , Poaceae/metabolismo , Plantas Tolerantes a Sal/metabolismo , Sódio/metabolismo , Vacúolos/metabolismo
6.
Cell Mol Neurobiol ; 42(4): 1049-1064, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-33258018

RESUMO

Histone deacetylases (HDACs) have been described to have both neurotoxic and neuroprotective roles, and partly, depend on its sub-cellular distribution. HDAC inhibitors have a long history of use in the treatment of various neurological disorders including epilepsy. Key role of HDACs in GABAergic neurotransmission, synaptogenesis, synaptic plasticity and memory formation was demonstrated whereas very less is known about their role in drug-resistant epilepsy pathologies. The present study was aimed to investigate the changes in the expression of HDACs, activity and its sub-cellular distribution in mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS) patients. For this study, surgically resected hippocampal tissue specimens of 28 MTLE-HS patients and 20 hippocampus from post-mortem cases were obtained. Real-time PCR was done to analyse the mRNA expression. HDAC activity and the protein levels of HDACs in cytoplasm as well as nucleus were measured spectrophotometrically. Further, sub-cellular localization of HDACs was characterized by immunofluorescence. Significant upregulation of HDAC1, HDAC2, HDAC4, HDAC5, HDAC6, HDAC10 and HDAC11 mRNA were observed in MTLE-HS. Alterations in the mRNA expression of glutamate and gamma-aminobutyric acid (GABA) receptor subunits have been also demonstrated. We observed significant increase of HDAC activity and nuclear level of HDAC1, HDAC2, HDAC5 and HDAC11 in the hippocampal samples obtained from patients with MTLE-HS. Moreover, we found altered cytoplasmic level of HDAC4, HDAC6 and HDAC10 in the hippocampal sample obtained from patients with MTLE-HS. Alterations in the level of HDACs could potentially be part of a dynamic transcription regulation associated with MTLE-HS. Changes in cytoplasmic level of HDAC4, 6 and 10 suggest that cytoplasmic substrates may play a crucial role in the pathophysiology of MTLE-HS. Knowledge regarding expression pattern and sub-cellular distribution of HDACs may help to devise specific HDACi therapy for epilepsy.


Assuntos
Epilepsia do Lobo Temporal , Epilepsia , Epilepsia/patologia , Epilepsia do Lobo Temporal/metabolismo , Hipocampo/metabolismo , Histona Desacetilases/metabolismo , Humanos , Imageamento por Ressonância Magnética , Esclerose/patologia
7.
Biochem Biophys Res Commun ; 569: 106-111, 2021 09 10.
Artigo em Inglês | MEDLINE | ID: mdl-34243065

RESUMO

ALS2, a product of the causative gene for familial amyotrophic lateral sclerosis (ALS) type 2, plays a pivotal role in the regulation of endosome dynamics by activating small GTPase Rab5 via its intrinsic guanine nucleotide-exchange factor activity. Previously, we have reported that the N-terminal region of ALS2 has crucial roles in its endosomal localization and self-oligomerization, both of which are indispensable for the cellular function of ALS2. The N-terminus of ALS2 contains the regulator of chromosome condensation 1-like domain (RLD), which is predicted to form a seven-bladed ß-propeller structure. Interestingly, the RLD is interrupted by the intrinsically disordered region (IDR), within which there are several amino acid residues which undergo phosphorylation. In this study, we sought to investigate as to whether and how the IDR as well as phosphorylation at either Ser483, Ser492 or Thr510 affect the intracellular localization and self-oligomerization of ALS2. All phospho- and dephospho-mimetic ALS2 mutants that were transiently expressed in HeLa cells were diffusely distributed throughout the cytosol with a partial localization to early endosomes. When expressed under Rac1-activating conditions, these mutants were localized to membrane ruffles as well as enlarged endosomes. Further, gel-filtration analysis revealed that these mutants primarily existed as a tetramer in cells. However, all these phenotypes were indistinguishable from those of wild-type ALS2. On the other hand, IDR-deleted ALS2 mutant was exclusively present in perinuclear aggregates colocalizing with the autophagy-related protein SQSTM1. Moreover, IDR-deleted ALS2 mutant formed an abnormally high molecular weight complex compared to wild-type ALS2. These results indicate that the IDR of ALS2 plays a crucial role not only in the regulation of intracellular localization but also in the self-oligomerization of ALS2 in cells, whereas phosphorylation of certain residues within the IDR exerts limited effects on such phenotypes.


Assuntos
Fatores de Troca do Nucleotídeo Guanina/química , Espaço Intracelular/metabolismo , Proteínas Intrinsicamente Desordenadas/química , Multimerização Proteica , Esclerose Lateral Amiotrófica/genética , Esclerose Lateral Amiotrófica/metabolismo , Western Blotting , Endossomos/metabolismo , Predisposição Genética para Doença/genética , Fatores de Troca do Nucleotídeo Guanina/genética , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Células HeLa , Humanos , Proteínas Intrinsicamente Desordenadas/genética , Proteínas Intrinsicamente Desordenadas/metabolismo , Microscopia de Fluorescência , Mutação , Fosforilação , Ligação Proteica , Transporte Proteico , Proteína Sequestossoma-1/metabolismo , Proteínas rab5 de Ligação ao GTP/metabolismo
8.
J Exp Bot ; 72(4): 1490-1505, 2021 02 24.
Artigo em Inglês | MEDLINE | ID: mdl-33170269

RESUMO

Very few of the >650 Proteaceae species in south-western Australia cope with the high calcium (Ca) levels in young, calcareous soils (soil indifferent); most are Ca sensitive and occur on nutrient-impoverished, acidic soils (calcifuge). We assessed possible control points for Ca transport across roots of two soil-indifferent (Hakea prostrata and Banksia prionotes) and two calcifuge (H. incrassata and B. menziesii) Proteaceae. Using quantitative X-ray microanalysis, we investigated cell-specific elemental Ca concentrations at two positions behind the apex in relation to development of apoplastic barriers in roots of plants grown in nutrient solution with low or high Ca supply. In H. prostrata, Ca accumulated in outer cortical cells at 20 mm behind the apex, but [Ca] was low in other cell types. In H. incrassata, [Ca] was low in all cells. Accumulation of Ca in roots of H. prostrata corresponded to development of apoplastic barriers in the endodermis. We found similar [Ca] profiles in roots and similar [Ca] in leaves of two contrasting Banksia species. Soil-indifferent Hakea and Banksia species show different strategies to inhabit calcareous soils: H. prostrata intercepts Ca in roots, reducing transport to shoots, whereas B. prionotes allocates Ca to specific leaf cells.


Assuntos
Proteaceae , Fósforo , Raízes de Plantas/química , Solo , Austrália Ocidental
9.
Glycobiology ; 30(2): 120-129, 2020 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-31584064

RESUMO

Galectin-13 (Gal-13) plays numerous roles in regulating the relationship between maternal and fetal tissues. Low expression levels or mutations of the lectin can result in pre-eclampsia. The previous crystal structure and gel filtration data show that Gal-13 dimerizes via formation of two disulfide bonds formed by Cys136 and Cys138. In the present study, we mutated them to serine (C136S, C138S and C136S/C138S), crystalized the variants and solved their crystal structures. All variants crystallized as monomers. In the C136S structure, Cys138 formed a disulfide bond with Cys19, indicating that Cys19 is important for regulation of reversible disulfide bond formation in this lectin. Hemagglutination assays demonstrated that all variants are inactive at inducing erythrocyte agglutination, even though gel filtration profiles indicate that C136S and C138S could still form dimers, suggesting that these dimers do not exhibit the same activity as wild-type (WT) Gal-13. In HeLa cells, the three variants were found to be distributed the same as with WT Gal-13. However, a Gal-13 variant (delT221) truncated at T221 could not be transported into the nucleus, possibly explaining why women having this variant get pre-eclampsia. Considering the normally high concentration of glutathione in cells, WT Gal-13 should exist mostly as a monomer in cytoplasm, consistent with the monomeric variant C136S/C138S, which has a similar ability to interact with HOXA1 as WT Gal-13.


Assuntos
Dissulfetos , Galectinas , Proteínas da Gravidez , Cristalografia por Raios X , Dissulfetos/química , Dissulfetos/metabolismo , Feminino , Galectinas/química , Galectinas/metabolismo , Células HeLa , Humanos , Oxirredução , Pré-Eclâmpsia/metabolismo , Gravidez , Proteínas da Gravidez/química , Proteínas da Gravidez/metabolismo , Domínios Proteicos , Relação Estrutura-Atividade
10.
Cell Mol Life Sci ; 76(3): 441-451, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30374521

RESUMO

MicroRNAs (miRNAs) are a class of endogenous small noncoding RNAs that participate in a majority of biological processes via regulating target gene expression. The post-transcriptional repression through miRNA seed region binding to 3' UTR of target mRNA is considered as the canonical mode of miRNA-mediated gene regulation. However, emerging evidence suggests that other regulatory modes exist beyond the canonical mechanism. In particular, the function of intranuclear miRNA in gene transcriptional regulation is gradually revealed, with evidence showing their contribution to gene silencing or activating. Therefore, miRNA-mediated regulation of gene transcription not only expands our understanding of the molecular mechanism underlying miRNA regulatory function, but also provides new evidence to explain its ability in the sophisticated regulation of many bioprocesses. In this review, mechanisms of miRNA-mediated gene transcriptional and post-transcriptional regulation are summarized, and the synergistic effects among these actions which form a regulatory network of a miRNA on its target are particularly elaborated. With these discussions, we aim to emphasize the importance of miRNA regulatory network on target gene regulation and further highlight the potential application of the network mode in the achievement of a more effective and stable modulation of the target gene expression.


Assuntos
Regulação da Expressão Gênica/genética , MicroRNAs/metabolismo , Sistemas de Liberação de Medicamentos , Inativação Gênica , Humanos , MicroRNAs/genética , Processamento de Proteína Pós-Traducional/genética , Ativação Transcricional/genética
11.
J Fish Dis ; 43(7): 791-799, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32449196

RESUMO

Koi herpesvirus (KHV) is an emerging pathogen of koi and common carp that causes a severe disease and mass mortality of infected fish. The KHV ORF72 protein is an important capsid protein that has been suggested to be a candidate for the development of diagnostic reagents and KHV vaccines. The purpose of this study was to clone and express the KHV ORF72 gene for further preparation of a specific monoclonal antibody (mAb) and to analyse cellular distribution of the viral protein. The mAb 3E1 could specifically recognize the expressed ORF72 protein of transfected cells by indirect immunofluorescence, and the antigenic site recognized by the mAb 3E1 was mapped to the region of N-terminal 124 residues of KHV ORF72. This mAb was further demonstrated to specifically detect the KHV-infected fish tissue by immunohistochemistry, thereby suggesting its high diagnostic potential. In addition, the cellular distribution analysis of the KHV ORF72 protein revealed that the region of amino acid residues 125-247 was related to mitochondrial localization and proliferation. Furthermore, a putative nuclear export signal (NES) of ORF72 at the residues 201-212 was confirmed on the basis of its function associated with NES activity.


Assuntos
Anticorpos Monoclonais/isolamento & purificação , Anticorpos Antivirais/isolamento & purificação , Doenças dos Peixes/imunologia , Infecções por Herpesviridae/veterinária , Herpesviridae/imunologia , Proteínas Virais/isolamento & purificação , Animais , Doenças dos Peixes/virologia , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/virologia , Distribuição Tecidual
12.
J Exp Bot ; 70(18): 4991-5002, 2019 09 24.
Artigo em Inglês | MEDLINE | ID: mdl-31106833

RESUMO

Salinity tolerance is associated with Na 'exclusion' from, or 'tissue tolerance' in, leaves. We investigated whether two contrasting chickpea genotypes, salt-tolerant Genesis836 and salt-sensitive Rupali, differ in leaf tissue tolerance to NaCl. We used X-ray microanalysis to evaluate cellular Na, Cl, and K concentrations in various cell types within leaflets and also in secretory trichomes of the two chickpea genotypes in relation to photosynthesis in control and saline conditions. TEM was used to assess the effects of salinity on the ultrastructure of chloroplasts. Genesis836 maintained net photosynthetic rates (A) for the 21 d of salinity treatment (60 mM NaCl), whereas A in Rupali substantially decreased after 11 d. Leaflet tissue [Na] was low in Genesis836 but had increased markedly in Rupali. In Genesis836, Na was accumulated in epidermal cells but was low in mesophyll cells, whereas in Rupali cellular [Na] was high in both cell types. The excessive accumulation of Na in mesophyll cells of Rupali corresponded to structural damage to the chloroplasts. Maintenance of photosynthesis and thus salinity tolerance in Genesis836 was associated with an ability to 'exclude' Na from leaflets and in particular from the photosynthetically active mesophyll cells, and to compartmentalize Na in epidermal cells.


Assuntos
Cicer/fisiologia , Células do Mesofilo/metabolismo , Tolerância ao Sal , Sódio/metabolismo , Cloro/metabolismo , Potássio/metabolismo
13.
Fish Shellfish Immunol ; 94: 373-380, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31533080

RESUMO

Retinoic acid-inducible gene-I (RIG-I) is a cytoplasmic viral RNA sensor that triggers the production of type I interferons (IFNs) and proinflammatory cytokines during viral infection. RIG-I gene has been identified previously in Japanese eel, Anguilla japonica. In the present study, we have characterized a novel isoform of RIG-I (designated as AjRIG-Ib) and its truncated variant (AjRIG-Ibv). The AjRIG-Ib encodes 940 amino acids (aa) consisting of two N-terminal caspase activation and recruitment domains (CARDs), a DEX(D/H) box RNA helicase domain, and a C-terminal regulatory domain (CTD). The AjRIG-Ibv encodes a protein of 843 aa, that shares similar structural organization with AjRIG-Ib, but lacking CTD. The gene expression analyses showed that AjRIG-Ib and AjRIG-Ibv were detectable in all tissues/organs examined, and AjRIG-Ib was the predominant form. The mRNA level of AjRIG-Ibv was upregulated rapidly at 8 h after the Poly I:C injection, and the significant increase of AjRIG-Ib was observed at 16 and 24 h post-injection (hpi). Laser confocal microscopy showed that AjRIG-Ib and AjRIG-Ibv were both located in cytoplasm. In addition, the overexpression of AjRIG-Ib or AjRIG-Ibv led to the increased activity of IFN promoter in transient transfection assay. Taken together, our results indicated that AjRIG-Ib and AjRIG-Ibv may play cooperative or somewhat complementary roles in coordinating the antiviral response in fish.


Assuntos
Anguilla/genética , Anguilla/imunologia , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Sequência de Aminoácidos , Animais , Proteínas de Peixes/química , Perfilação da Expressão Gênica/veterinária , Filogenia , Poli I-C/farmacologia , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Alinhamento de Sequência/veterinária
14.
Int J Phytoremediation ; 21(1): 4-13, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30638050

RESUMO

A field experiment in which a hyperaccumulator (Arabis alpina) was intercropped with winter crop (Vicia faba), was conducted to understand effect of the root exudates on the content and accumulated amounts, sub-cellular distribution of Cd and Pb of the intercropped plants during the ripening period of V. faba (120 d after sowing). The results showed that contents of soluble sugars exuded from the roots of intercropped A. alpina were 67.6% less than that of the monocropped plant, whereas contents of free amino acids was 57.9% greater. The total contents of organic acids exuded from roots of intercropped A. alpina and V. faba were 578.8% and 37.8% greater than that of monocropped plants, respectively. The contents of tartaric acid and malic acid exuded by roots of intercropped A. alpina were greater 31.9 times and 15.9 times than those of monocropped A. alpina, respectively. The contents and accumulated amounts of Cd and Pb in intercropped A. alpina were greater than those of monocropped A. alpina. The contents of Pb bound to organic matter in cell walls, cytoplasm and organelles of intercropped plants were greater than those of monocropped plants. These results demonstrate that increases in accumulated amounts of Pb and Cd caused by intercropping were closely related to migration of Cd and Pb in plants mediated by the composition and content of the root exudates.


Assuntos
Arabis , Vicia faba , Biodegradação Ambiental , Cádmio , Chumbo , Raízes de Plantas , Triticum
15.
Planta ; 248(1): 49-68, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29564629

RESUMO

MAIN CONCLUSION: Salicylic acid alleviates lead toxicity in Brassica juncea (L.) by promoting growth under non-stress and activating stress-defense mechanism (s) under lead stress conditions. It also boosts the ascorbate-glutathione cycle and thus helps in minimizing oxidative and DNA damage. Brassica juncea plants were exposed to different concentrations (0, 500, 1000 and 2000 mg kg-1) of lead (Pb) and subsequently sprayed with 0.5 mM of salicylic acid (SA) to check for morphological and leaf gas exchange parameters like transpiration rate (E), stomatal conductance (GH2O), net photosynthetic rate (A) and maximum quantum yield of PS II (Fv/Fm). Leaf epidermis by scanning electron microscopy (SEM), enzymatic and non-enzymatic components of ascorbate-glutathione (AsA-GSH) cycle, DNA damage by comet assay, lipid peroxidation and endogenous SA quantification by HPLC were analyzed. Lead accumulation in root, shoot and its sub-cellular distribution ratio (SDR) and localization was also determined using atomic absorption spectroscopy (AAS) and rhodizonate-dye staining method, respectively. Results revealed that notable amount of Pb was accumulated in root and shoot in dose-dependent manner which significantly (P ≤ 0.05) posed the toxicity on the majority of morphological parameters, structural integrity of epidermal and guard cells, photosynthetic pigments, malondialdehyde (MDA) and H2O2 content. Notable decrease in leaf gas exchange parameters, Fv/Fm, poor performance of AsA-GSH cycle and striking amount of DNA damage, was found as well. However, SA revoked Pb toxicity to a great extent by promoting growth, chlorophyll content, improving the A, Fv/Fm, boosting the overall performance of AsA-GSH cycle and by lessening the DNA damage.


Assuntos
Chumbo/toxicidade , Mostardeira/efeitos dos fármacos , Ácido Salicílico/farmacologia , Dano ao DNA/efeitos dos fármacos , Relação Dose-Resposta a Droga , Chumbo/análise , Peroxidação de Lipídeos/efeitos dos fármacos , Microscopia Eletrônica de Varredura , Fotossíntese/efeitos dos fármacos , Complexo de Proteína do Fotossistema II/efeitos dos fármacos , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Folhas de Planta/ultraestrutura , Raízes de Plantas/química , Brotos de Planta/química , Estômatos de Plantas/efeitos dos fármacos , Transpiração Vegetal/efeitos dos fármacos
16.
Fish Shellfish Immunol ; 81: 374-382, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30016685

RESUMO

Myeloid differentiation factor 88 (MyD88) is a key adaptor protein required for the signaling of all Toll-like receptors except TLR3, which results to the interaction of activated TLR complexes via C-terminal TIR domain and the binding of downstream kinase via N-terminal death domain. In this study, the MyD88 gene from the Japanese eel (Anguilla japonica) was identified. The open reading frame of AjMyD88 was 918 bp in length, encoding a protein composed of conserved N-terminal death domain and C-terminal TIR domain, respectively. Multiple alignment revealed highly conserved sites across all examined vertebrate lineages in death and TIR domains. Site-directed mutagenesis and luciferase analysis revealed that the W78A, L91A and L95A mutations in death domain had modest impairment of their ability in activating NF-κB promoter. The expression level of AjMyD88 was investigated by real-time PCR in response to poly I:C stimulation and Edwardsiella tarda infection. Significantly increased MyD88 expression was observed at early phase in all tested tissues/organs in response to E. tarda infection and slight increase was detected in intestine and gill at 16 hpi and in head kidney, spleen and liver at 24 hpi after poly I:C stimulation. Immunofluorescence staining revealed that AjMyD88 is present as condensed forms in the cytoplasm. Taken together, sequence characterization, gene expression and cellular distribution data obtained in this study suggest that AjMyD88, similar to its mammalian ortholog, plays an important role in eel immune response against bacteria.


Assuntos
Anguilla/genética , Fator 88 de Diferenciação Mieloide/genética , Sequência de Aminoácidos , Anguilla/imunologia , Anguilla/microbiologia , Animais , DNA Complementar/genética , Edwardsiella tarda , Fatores Imunológicos/farmacologia , Fator 88 de Diferenciação Mieloide/imunologia , Filogenia , Poli I-C/farmacologia , Alinhamento de Sequência
17.
Biometals ; 31(1): 17-28, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-29188540

RESUMO

Plants suffer from combined stress of sulfur deficiency and cadmium toxicity in some agricultural lands. However, little is known about the reaction in plants, such as responses in antioxidant enzymes and non-protein thiol compounds, to such combined stress. Therefore, in this study, four treatments, S-sufficiency (TS-Cd), S-deficiency (T-S-Cd), Cd stress (TS+Cd) and combined stress of S-deficiency and Cd stress (T-S+Cd), were set up to investigate (1) the effects of sulfur deficiency or sulfur sufficiency on Cd toxicity to kidney bean cultivar seedlings and the related mechanisms, and (2) the responses of two kidney bean cultivars to combined stress of S-deficiency and Cd-tolerance. The results showed significant increases in hydrogen peroxide (H2O2) and malondialdehyde contents and significant increases in antioxidant enzyme (superoxide dismutase, catalase, peroxidase, and glutathione S-transferase) activities and non-protein thiol compounds (non-protein thiols, reduced glutathione, phytochelatins) synthesis in the plants in TS+Cd and T-S+Cd. On the tissue level, higher proportion of Cd was found to be immobilized/deposited in roots, while on the sub-cell level, higher proportion of Cd was located in cell walls and vacuole fractions with lower in cell organelles. Taken together, the results indicated that Cd detoxification was achieved by the two kidney bean cultivars through antioxidant enzyme activation, non-protein thiol compound synthesis and sub-cellular compartmentalization. In addition, the results indicated that sufficient S supply helped to relieve Cd toxicity, which is of special significance for remediation or utilization of Cd-contaminated soils as S is a plant essential nutrient.


Assuntos
Adaptação Fisiológica/genética , Cádmio/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , Phaseolus/efeitos dos fármacos , Proteínas de Plantas/genética , Enxofre/deficiência , Catalase/genética , Catalase/metabolismo , Parede Celular/efeitos dos fármacos , Parede Celular/metabolismo , Glutationa/metabolismo , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Peróxido de Hidrogênio/agonistas , Peróxido de Hidrogênio/metabolismo , Estresse Oxidativo , Phaseolus/genética , Phaseolus/metabolismo , Fitoquelatinas/metabolismo , Células Vegetais/efeitos dos fármacos , Células Vegetais/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/metabolismo , Especificidade da Espécie , Estresse Fisiológico , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Vacúolos/efeitos dos fármacos , Vacúolos/metabolismo
18.
Molecules ; 23(11)2018 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-30424011

RESUMO

Eosinophils are present in tissues, such as the respiratory tract, spleen, lymph nodes and blood vessels. The significant presence of eosinophils in these tissues are associated with various diseases, including asthma, allergies, acute myeloid leukemia, etc. Charcot-Leyden crystal protein/galectin-10 is overexpressed in eosinophils and has also been identified in basophils and macrophages. In human body, this protein could spontaneously form Charcot-Leyden crystal in lymphocytes or in the lysates of lymphocytes. At present, the role of Charcot-Leyden crystal protein/galectin-10 in lymphocytes is not fully understood. This review summarizes research progress on Charcot-Leyden crystal protein/galectin-10, with emphasis on its history, cellular distributions, relations to diseases, structures and ligand binding specificity.


Assuntos
Galectinas/química , Galectinas/metabolismo , Glicoproteínas/química , Glicoproteínas/metabolismo , Lisofosfolipase/química , Lisofosfolipase/metabolismo , Pesquisa , Animais , Cristalização , Suscetibilidade a Doenças , Galectinas/genética , Glicoproteínas/genética , Humanos , Ligantes , Linfócitos/metabolismo , Lisofosfolipase/genética , Regiões Promotoras Genéticas , Ligação Proteica , Transporte Proteico , Coloração e Rotulagem , Relação Estrutura-Atividade
19.
NMR Biomed ; 30(4)2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27119601

RESUMO

Iron is an essential trace element involved in a variety of biological mechanisms in the human body. Disturbances of iron homeostasis have been observed in several inflammatory and degenerative diseases, which have raised strong interest in non-invasive iron mapping techniques. Numerous MRI techniques have been proposed so far, mostly based on the field changes induced by the magnetic properties of iron. Each of these approaches has a specific sensitivity for iron and its microstructural environment. Quantitative susceptibility mapping is the latest development and provides a direct measure of bulk susceptibility. However, field changes induced by iron are not always directly related to the concentration of iron, but rather reflect the structure of iron compounds and its cellular distribution. This review provides an overview of the most relevant iron compounds in the human body, their magnetic properties and their cellular distribution. In addition, MRI methods based on direct or indirect susceptibility changes are presented and discussed with respect to technical aspects and clinical applicability. Copyright © 2016 John Wiley & Sons, Ltd.


Assuntos
Química Encefálica , Interpretação de Imagem Assistida por Computador/métodos , Ferro/análise , Ferro/química , Espectroscopia de Ressonância Magnética/métodos , Imagem Molecular/métodos , Animais , Biomarcadores/análise , Biomarcadores/química , Encéfalo/anatomia & histologia , Humanos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Distribuição Tecidual
20.
J Biol Inorg Chem ; 22(8): 1295-1304, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29043454

RESUMO

Oxaliplatin is the backbone of chemotherapy for advanced colorectal cancer and undergoes clinical trials for treatment of other tumour entities. However, acquired resistance is a major hurdle. Confocal microscopy is a useful tool to get an insight into the mechanisms of resistance but it requires fluorescent compounds. This work describes the synthesis of the novel oxaliplatin derivative (CFDA-oxPt) featuring 5(6)-carboxyfluorescein diacetate and evaluation of its applicability for the investigation of oxaliplatin resistance using imaging techniques. CFDA-oxPt was somewhat less cytotoxic than oxaliplatin in sensitive colorectal cancer cells, with EC50 values of 26 and 5.8 µM, respectively. Nevertheless, the potency of the novel complex was significantly decreased to the EC50 of 711.2 µM in oxaliplatin-resistant cells, as was the case for oxaliplatin (EC50 = 81 µM). After incubation, both nuclear and cytosolic localisation was observed. Over time CFDA-oxPt concentrated near the cell membrane and in the vesicular structures, in contrast to the platinum-free label, which was rapidly excreted. These findings suggest that CFDA-oxPt can be used to study oxaliplatin resistance and open the route to new fluorophore-tethered oxaliplatin derivatives.


Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Resistencia a Medicamentos Antineoplásicos , Corantes Fluorescentes/química , Imagem Molecular , Compostos Organoplatínicos/química , Compostos Organoplatínicos/farmacologia , Antineoplásicos/metabolismo , Transporte Biológico , Linhagem Celular Tumoral , Fluoresceínas/química , Humanos , Compostos Organoplatínicos/metabolismo , Oxaliplatina
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