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1.
Nat Commun ; 11(1): 862, 2020 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-32054841

RESUMO

Complex hierarchical structure governs emergent properties in biopolymeric materials; yet, the material processing involved remains poorly understood. Here, we investigated the multi-scale structure and composition of the mussel byssus cuticle before, during and after formation to gain insight into the processing of this hard, yet extensible metal cross-linked protein composite. Our findings reveal that the granular substructure crucial to the cuticle's function as a wear-resistant coating of an extensible polymer fiber is pre-organized in condensed liquid phase secretory vesicles. These are phase-separated into DOPA-rich proto-granules enveloped in a sulfur-rich proto-matrix which fuses during secretion, forming the sub-structure of the cuticle. Metal ions are added subsequently in a site-specific way, with iron contained in the sulfur-rich matrix and vanadium coordinated by DOPA-catechol in the granule. We posit that this hierarchical structure self-organizes via phase separation of specific amphiphilic proteins within secretory vesicles, resulting in a meso-scale structuring that governs cuticle function.


Assuntos
Materiais Revestidos Biocompatíveis/química , Metaloproteínas/química , Mytilus edulis/química , Estruturas Animais/anatomia & histologia , Estruturas Animais/química , Estruturas Animais/ultraestrutura , Animais , Di-Hidroxifenilalanina/química , Imageamento Tridimensional , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Mytilus edulis/anatomia & histologia , Mytilus edulis/ultraestrutura , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Vesículas Secretórias/química , Vesículas Secretórias/ultraestrutura
2.
Food Chem ; 303: 125399, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31470274

RESUMO

It is still a challenge to solve the matrix interferences in veterinary drug residue analysis. In this study, we reported a thin layer chromatography (TLC)-high-performance liquid chromatography (HPLC) method for determining total florfenicol (FF) residues, expressed as florfenicol amine (FFA), in porcine edible tissues. The tissue homogenate were acid-hydrolyzed to liberate the bound residues and convert them into FFA. The hydrolysates were washed with ethyl acetate and subsequently extracted with ethyl acetate under alkaline conditions. The supernatants were concentrated through evaporation, defatted with hexane, purified by TLC and analyzed by HPLC at 225 nm. The optimal developing solvent for TLC purification was ethyl acetate-acetone-ammonium hydroxide mixtures (2:8:0.5, v/v/v). The method was fully validated according to decision 2002/657/EC, and could be used for the routine monitoring of FF residues in pig. TLC showed excellent purification efficiency, and was expected to solve the matrix interferences in veterinary drug residue analysis.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Cromatografia em Camada Delgada/métodos , Resíduos de Drogas/análise , Tianfenicol/análogos & derivados , Drogas Veterinárias/análise , Estruturas Animais/química , Animais , Cromatografia Líquida/métodos , Carne/análise , Suínos , Tianfenicol/análise
3.
Biol Bull ; 237(1): 1-15, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31441702

RESUMO

The pen, or gladius, of the squid is an internalized shell. It serves as a site of attachment for important muscle groups and as a protective barrier for the visceral organs. The pen's durability and flexibility are derived from its unique composition of chitin and protein. We report the characterization of the structure, development, and composition of pens from Doryteuthis pealeii. The nanofibrils of the polysaccharide ß-chitin are arranged in an aligned configuration in only specific regions of the pen. Chitin is secreted early in development, enabling us to characterize the changes in pen morphology prior to hatching. The chitin and proteins are assembled in the shell sac surrounded by fluid that has a significantly different ionic composition from squid plasma. Two groups of proteins are associated with the pen: those on its surface and those embedded within the pen. Only 20 proteins are identified as embedded within the pen. Embedded proteins are classified into six groups, including chitin associated, protease, protease inhibitors, intracellular, extracellular matrix, and those that are unknown. The pen proteins share many conserved domains with proteins from other chitinous structures. We conclude that the pen is one of the least complex, load-bearing, chitin-rich structures currently known and is amenable to further studies to elucidate natural construction mechanisms using chitin and protein.


Assuntos
Quitina/metabolismo , Decapodiformes/anatomia & histologia , Proteínas/metabolismo , Estruturas Animais/anatomia & histologia , Estruturas Animais/química , Estruturas Animais/crescimento & desenvolvimento , Animais , Decapodiformes/química , Decapodiformes/crescimento & desenvolvimento
4.
Neotrop Entomol ; 48(6): 1030-1038, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31456168

RESUMO

Chemical compounds present on the cuticle of social insects are important in communication, as they are used in recognition of nestmates and sexual partners as well as in caste distinction, varying according to several factors, such as genetic and environmental. In this context, some studies have explored the cuticular chemical profile as a tool for assessing intra- and interspecific differences in social insects, although few studies have investigated this in social wasps. This study aimed to assess the differences in cuticular chemical profiles among different geographic samples of the wasp Mischocyttarus consimilis Zikán. Our hypothesis was that environmental factors are decisive to compose the cuticular chemical profiles of colonies of these social wasps and that there are differences regarding the geographic distribution among colonies. We used Fourier Transform Infrared-Photoacoustic Spectroscopy (FTIR-PAS) to assess the chemical profiles of samples. Our results show that despite there are differences between the cuticular chemical composition of the wasps' samples from different populations, there is no significant correlation compared to the spatial distribution of the colonies nor with the environment. Thus, our hypothesis was refuted, and we can infer that in this species neither exogenous nor genetic factors stand out to differentiate the chemical signature of their colonies, but a combination of both.


Assuntos
Estruturas Animais/química , Variação Biológica da População , Espectroscopia de Infravermelho com Transformada de Fourier , Vespas/química , Animais , Brasil , Geografia
5.
Int J Biol Macromol ; 135: 959-968, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31136752

RESUMO

The structure and properties of natural sheep casing and collagen films with various crosslinking treatments have been investigated in detail to develop satisfied artificial casings prepared from collagen. The sheep casing consists of large number of thick collagen fibers oriented at ±45° from longitudinal direction with high-density interwoven network structure. The structural feature of sheep casing gave the special mouthfeel of 'cracking bite' of sausages. Whereas, layered structure filled with fine collagen fibrils and large gaps in collagen film results in poor mechanical properties and higher swelling ratio in water. Furthermore, a degree of denaturation of collagen during extraction process also lead to poor mechanical properties. After glutaraldehyde (GTA) and dehydrothermal (DHT) treatments, the formation of crosslinking improved mechanical properties of collagen films significantly and the tensile strength and tensile modulus increased more than three times compared with those of untreated collagen film in wet before and after boiling. The swelling ratio of treated collagen films also decreased dramatically. No obvious effects on denaturation of collagen film after GTA treatment, but the degree of denaturation of DHT treated collagen film increased slightly.


Assuntos
Produtos Biológicos/química , Fenômenos Químicos , Colágeno/química , Membranas Artificiais , Estrutura Molecular , Estruturas Animais/química , Animais , Bovinos , Reagentes para Ligações Cruzadas/química , Fenômenos Mecânicos , Reologia , Ovinos
6.
Theranostics ; 9(4): 932-944, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30867807

RESUMO

Detailed spatio-temporal information on drug distribution in organs is of paramount importance to assess drug clinically-relevant properties and potential side-effects. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) as a label-free and sensitive imaging modality provides an additional means of accurately visualizing drug and its metabolites distributions in tissue sections. However, technical limitations, complex physiochemical environment of surface and low abundance of target drugs make quantitative MALDI imaging of drug and its metabolites quite challenging. Methods: In this study, an internal standard correction strategy was applied for quantitative MALDI imaging of tetrandrine in multiple organs of rats including lung, liver, kidney, spleen, and heart. The feasibility and reliability of the developed quantitative MSI method were validated by conventional liquid chromatography-tandem MS (LC-MS/MS) analysis, and the two methods showed a significant correlation. Results: The quantitative MALDI imaging method met the requirements of specificity, sensitivity and linearity. Tissue-specific spatio-temporal distribution patterns of tetrandrine in different organs were revealed after intravenous administration in the rat. Moreover, demethylated metabolite was detected in liver tissues. Conclusions: The current work illustrates that quantitative MALDI imaging provides an alternative means of accurately addressing the problem of drug and its metabolites distribution in tissues, complementary to traditional LC-MS/MS of tissue homogenates and whole-body autoradiography (WBA). Quantitative spatio-chemical information obtained here can improve our understanding of pharmacokinetics (PK), pharmacodynamics (PD), and potential transient toxicities of tetrandrine in organs, and possibly direct further optimization of drug properties to reduce drug-induced organ toxicity.


Assuntos
Estruturas Animais/química , Antimaláricos/farmacocinética , Antineoplásicos Fitogênicos/farmacocinética , Benzilisoquinolinas/farmacocinética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Antimaláricos/administração & dosagem , Antineoplásicos Fitogênicos/administração & dosagem , Benzilisoquinolinas/administração & dosagem , Ratos , Análise Espaço-Temporal
7.
Mar Pollut Bull ; 137: 152-156, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30503421

RESUMO

The purpose of this study is to establish the presence of POPs in the Bering flounder (Hippoglossoides robustus) from the Sea of Okhotsk (North-West Pacific). Concentration of OCPs (α-HCH, ß-HCH, γ-HCH, o,p'-DDT, p,p'-DDT, o,p'-DDD, p,p'-DDD, o,p'-DDE and p,p'-DDE) and PCBs (28, 52, 155, 101, 118, 143, 153, 138, 180, 207) in samples were measured by GC-MS and GC-ECD. The mean OCP concentrations in flounder from East and South areas of the Sea of Okhotsk were 99.8 ±â€¯125.4 and 53.6 ±â€¯40.5 ng/g lipid, respectively; PCB congeners - 112 ±â€¯94.2 and 88.8 ±â€¯50.8 ng/g lipid, respectively. POPs in fish tissue decreased in the order: PCBs > HCHs > DDTs. Our results indicate that consumers will have no health risk due to fish consumption from Sea of Okhotsk. OCP and PCB levels in the Sea of Okhotsk may be considered as background level for the North Pacific.


Assuntos
Estruturas Animais/química , Linguado/metabolismo , Hidrocarbonetos Clorados/química , Praguicidas/química , Bifenilos Policlorados/química , Poluentes Químicos da Água/química , Estruturas Animais/metabolismo , Animais , Cromatografia Gasosa-Espectrometria de Massas , Hidrocarbonetos Clorados/metabolismo , Praguicidas/metabolismo , Bifenilos Policlorados/metabolismo , Poluentes Químicos da Água/metabolismo
8.
Zoolog Sci ; 35(6): 535-547, 2018 12 04.
Artigo em Inglês | MEDLINE | ID: mdl-30520360

RESUMO

Most ascidian species settle on underwater substrates during a short free-swimming tadpole larval period. During this process, "rapid adhesion" occurs on adhesive papillae located at the anterior region of the cephalenteron. Settled and transformed ascidians subsequently expand the attachment area by "slow adhesion" with ampullae. In the present study, we attempted to identify the ultrastructures related to the adhesion process and adhesive materials in the ascidian tunic and to elucidate the biological function of vanadium in adhesion. We focused on an adhesive organ named the adhesive projection, which is newly generated by the adhered tunic to enlarge the bonding area between ascidian and substrate. Based on its structure and the presence of vanadiumcontaining blood cells, the adhesive projection was considered to be a large tunic vessel. At the adhered tunic, eosinophilic regions and migrated tunic cells were observed, but metal deposition was not detected. We speculate that the eosinophilic materials were components of the adhesive glue, and these are likey produced in epithelial cells, tunic cells, or both. Furthermore, using imaging mass spectrometry, we identified eight tunic-specific molecules as glue candidates.


Assuntos
Estruturas Animais/química , Urocordados/fisiologia , Estruturas Animais/fisiologia , Animais , Epiderme , Espectrometria de Massas
9.
J R Soc Interface ; 15(148)2018 11 14.
Artigo em Inglês | MEDLINE | ID: mdl-30429263

RESUMO

Scarab beetles (Coleoptera: Scarabaeidae) can exhibit striking colours produced by pigments and/or nanostructures. The latter include helicoidal (Bouligand) structures that can generate circularly polarized light. These have a cryptic evolutionary history in part because fossil examples are unknown. This suggests either a real biological signal, i.e. that Bouligand structures did not evolve until recently, or a taphonomic signal, i.e. that conditions during the fossilization process were not conducive to their preservation. We address this issue by experimentally degrading circularly polarizing cuticle of modern scarab beetles to test the relative roles of decay, maturation and taxonomy in controlling preservation. The results reveal that Bouligand structures have the potential to survive fossilization, but preservation is controlled by taxonomy and the diagenetic history of specimens. Further, cuticle of specific genus (Chrysina) is particularly decay-prone in alkaline conditions; this may relate to the presence of certain compounds, e.g. uric acid, in the cuticle of these taxa.


Assuntos
Estruturas Animais , Besouros , Fósseis/ultraestrutura , Nanoestruturas , Pigmentação , Estruturas Animais/química , Estruturas Animais/ultraestrutura , Animais , Besouros/química , Besouros/ultraestrutura , Nanoestruturas/química , Nanoestruturas/ultraestrutura
10.
J Agric Food Chem ; 66(44): 11531-11543, 2018 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-30345762

RESUMO

Anthocyanins (ACNs) are promising health-enhancing phenolic compounds. We focus on ACN animal tissue bioavailability to provide an evidentiary link between tissue ACNs and their associated health properties. We performed a systematic review of electronic libraries; 279 results were retrieved, and 13 publications met inclusion criteria. Extracted information included animal model employed, administration route, doses, analysis method, and ACN concentration values in tissues. Total ACN concentrations were detected in mice kidney (2.17 × 105 pmol/g), liver (1.73 × 105 pmol/g), heart (3.6 × 103 pmol/g), and lung (1.16 × 105 pmol/g); and in pig brain (6.08 × 103 pmol/g). ACNs showed a predominance of parent ACNs in long-term experiments versus an ACN metabolite predominance in short-term experiments. ACNs detected in animal tissues, such as cyanidin-3-glucoside, suggest it may have an important role in human health. This information could be useful to determine proper ACN-intake biomarkers in biological samples in futures studies.


Assuntos
Antocianinas/metabolismo , Extratos Vegetais/metabolismo , Estruturas Animais/química , Estruturas Animais/metabolismo , Animais , Antocianinas/química , Disponibilidade Biológica , Humanos , Extratos Vegetais/química , Suínos
11.
J R Soc Interface ; 15(147)2018 10 24.
Artigo em Inglês | MEDLINE | ID: mdl-30355807

RESUMO

Marine mussels (Mytilus spp.) attach to a wide variety of surfaces underwater using a network of byssal threads, each tipped with a protein-based adhesive plaque that uses the surrounding seawater environment as a curing agent. Plaques undergo environmental post-processing, requiring a basic seawater pH be maintained for up to 8 days for the adhesive to strengthen completely. Given the sensitivity of plaques to local pH conditions long after deposition, we investigated the effect of other aspects of the seawater environment that are known to vary in nearshore habitats on plaque curing. The effect of seawater temperature, salinity and dissolved oxygen concentration were investigated using tensile testing, atomic force microscopy and amino acid compositional analysis. High temperature (30°C) and hyposalinity (1 PSU) had no effect on adhesion strength, while incubation in hypoxia (0.9 mg l-1) caused plaques to have a mottled coloration and prematurely peel from substrates, leading to a 51% decrease in adhesion strength. AFM imaging of the plaque cuticle found that plaques cured in hypoxia had regions of lower stiffness throughout, indicative of reductions in DOPA cross-linking between adhesive proteins. A better understanding of the dynamics of plaque curing could aid in the design of better synthetic adhesives, particularly in medicine where adhesion must take place within wet body cavities.


Assuntos
Adesividade , Estruturas Animais/química , Di-Hidroxifenilalanina/química , Di-Hidroxifenilalanina/fisiologia , Mytilus/fisiologia , Oxigênio/química , Animais , Microscopia de Força Atômica
12.
Theranostics ; 8(15): 4033-4049, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30128034

RESUMO

Rationale: Successful treatment of pancreatic cancer remains a challenge due to desmoplasia and prevalence of KRAS mutation. While hedgehog (Hh) ligand levels are upregulated in pancreatic cancer cells and contribute to desmoplasia, there is significant downregulation of tumor suppressor let-7b, which targets mutant KRAS, C-MYC and several other genes involved in pancreatic cancer progression, invasion, and metastasis. We recently explored combination therapy of GDC-0449 (Hh inhibitor) and let-7b mimic using poly(ethylene glycol)-block-poly(2-methyl-2-carboxyl-propylene carbonate-graft-dodecanol-graft-tetraethylenepentamine) (PEG-b-PCC-g-DC-g-TEPA) micelles in pancreatic tumor mouse model. Here, our objective was to determine the biodistribution (BD), pharmacokinetics (PK), therapeutic efficacy and toxicity of this micellar formulation. Methods: We determined the PK of micelles encapsulating Cy5.5-let-7b and GDC-0449 following intravenous injection in orthotopic pancreatic tumor-bearing NSG mice at doses of 2 mg/kg and 10 mg/kg, respectively. Mice were scanned for fluorescence by IVIS to determine the biodistribution of Cy5.5-let-7b at the whole-body level, and its concentration in plasma and major organs was determined by measuring fluorescence using a fluorimeter and by real-time RT-PCR. GDC-0449 concentration was determined by LC/MS/MS. Therapeutic efficacy and toxicity of the micellar formulation of let-7b and GDC-0449 was also determined after two weeks of treatment. Results: The use of a micellar formulation markedly prolonged the elimination half-life (t1/2, e) of Cy5.5-let-7b in plasma from 0.49 ± 0.19 h to 2.65 ± 0.46 h and increased the area-under-the-curve (AUC 0-∞ ) by 7-fold, while t1/2,e and AUC 0-∞ of GDC-0449 were increased by 1.78-fold and 3.2-fold, respectively. The micelles significantly decreased the clearance of both encapsulated let-7b mimic and GDC-0449 compared to the emulsion formulation. Compared to the emulsion counterpart, the micellar formulation elevated the delivery of Cy5.5-let-7b and GDC-0449 to the orthotopic pancreatic tumor tissue by 7.8- and 4.2-fold, respectively. Furthermore, there was a significant reduction in tumor volume and negligible systemic toxicity as evident by hematological parameters and histological evaluation. Conclusion: PEG-b-PCC-g-DC-g-TEPA micelles carrying GDC-0449 and let-7b mimic have great potential to improve drug delivery for pancreatic cancer treatment.


Assuntos
Antineoplásicos/farmacologia , Antineoplásicos/farmacocinética , Portadores de Fármacos/administração & dosagem , Micelas , MicroRNAs/farmacologia , MicroRNAs/farmacocinética , Neoplasias Pancreáticas/tratamento farmacológico , Anilidas/farmacocinética , Anilidas/farmacologia , Estruturas Animais/química , Animais , Carbocianinas/farmacocinética , Cromatografia Líquida , Modelos Animais de Doenças , Fluorometria , Camundongos , Imagem Óptica , Neoplasias Pancreáticas/patologia , Plasma/química , Piridinas/farmacocinética , Piridinas/farmacologia , Espectrometria de Massas em Tandem , Resultado do Tratamento , Imagem Corporal Total
13.
Rapid Commun Mass Spectrom ; 32(15): 1257-1262, 2018 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-29777557

RESUMO

RATIONALE: Baleen plates are anatomical structures composed of inert tissue that hang from the upper jaw in mysticetes. Baleen plates may differ in size and in coloration between different segments of the filtering row or between sides of the mouth. Concern has been raised that variation in baleen plate characteristics may reflect dissimilar structural composition and growth rates liable to affect stable isotope ratios and their oscillation patterns. METHODS: We measured stable carbon (δ13 C values) and nitrogen (δ15 N values) isotope ratios at intervals of 1 cm along the longitudinal axis of six baleen plates collected from different positions along the mouth of a fin whale. All samples were analysed using a continuous flow isotope ratio mass spectrometer. Generalized additive models were fitted to the data from each baleen plate and the results of the models were compared visually. RESULTS: A total of 206 samples were analysed. Visually, all baleen plates presented nearly identical oscillations, independent of the position or the coloration of the baleen plate. However, the variation in δ13 C and δ15 N values occurring between the different baleen plates was higher in the segments of oscillations exhibiting steeper slopes. CONCLUSIONS: Differences in size between plates in an individual are due to differential erosion rates according to their position in the mouth. Therefore, the position of sampling along the baleen plate row should not be a reason for concern when conducting stable isotope studies.


Assuntos
Estruturas Animais/química , Isótopos de Carbono/análise , Baleia Comum/fisiologia , Boca/química , Isótopos de Nitrogênio/análise , Animais , Comportamento Alimentar , Masculino , Espectrometria de Massas
14.
Environ Int ; 114: 212-218, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29522985

RESUMO

Persistent organic pollutants (POPs) are long-range transported to the Arctic via atmospheric and oceanic currents, where they biomagnify to high concentrations in the tissues of apex predators such as polar bears (Ursus maritimus). A major concern of POP exposure is their physiological effects on vital organ-tissues posing a threat to the health and survival of polar bears. Here we examined the relationship between selected POPs and baculum bone mineral density (BMD) in the East Greenland and seven Canadian subpopulations of polar bears. BMD was examined in 471 bacula collected between years 1996-2015 while POP concentrations in adipose tissue were determined in 67-192 of these individuals collected from 1999 to -2015. A geographical comparison showed that baculum BMD was significantly lowest in polar bears from East Greenland (EG) when compared to Gulf of Boothia (GB), Southern Hudson (SH) and Western Hudson (WH) Bay subpopulations (all p < 0.05). The calculation of a T-score osteoporosis index for the EG subpopulation using WH bears as a reference group gave a T-score of -1.44 which indicate risk of osteopenia. Concentrations of ΣPCB74 (polychlorinated biphenyls), ΣDDT3 (dichlorodiphenyltrichloroethanes), p,p'-DDE (dichlorodiphenyldichloroethylene), ΣHCH3 (hexachlorohexane) and α-HCH was significantly highest in EG bears while ΣPBDE (polybrominated diphenyl ethers), BDE-47 and BDE-153 was significantly highest in SH bears (all p < 0.04). Statistical analyses of individual baculum BMD vs. POP concentrations showed that BMD was positively correlated with ΣPCB74, CB-153, HCB (hexachlorobenzene), ΣHCH, ß-HCH, ClBz (chlorobenzene), ΣPBDE and BDE-153 (all p < 0.03). In conclusion, baculum density was significantly lowest in East Greenland polar bears despite the positive statistical correlations of BMD vs. POPs. Other important factors such as nutritional status, body mass and body condition was not available for the statistical modelling. Since on-going environmental changes are known to affect these, future studies need to incorporate nutritional, endocrine and genetic parameters to further understand how POP exposure may disrupt bone homeostasis and affect baculum BMD across polar bear subpopulations.


Assuntos
Densidade Óssea/fisiologia , Osso e Ossos/química , Poluentes Ambientais/análise , Bifenilos Policlorados/análise , Ursidae , Estruturas Animais/química , Animais , Masculino , Pênis/química
15.
Sci Rep ; 8(1): 723, 2018 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-29335509

RESUMO

Interleukin 1 is a pleiotropic cytokine that mediates diverse functions through its receptor, type I interleukin 1 receptor (IL-1R1). Most previous studies have focused on the expression and function of IL-1R1 in immune cells. Here we performed a comprehensive mapping of IL-1R1 distribution in multiple peripheral tissues using our IL-1R1 reporter (IL-1R1GR/GR) mice. This method yielded the highest sensitivity of in situ detection of IL-1R1 mRNA and protein. Besides validating previously reported IL-1R1 expression in the endocrine tissues including pituitary and pancreas, our results refuted previously reported exclusive IL-1R1 expression in neurons of the spinal cord dorsal horn and dorsal root ganglia (DRG). Instead, IL-1R1 expression was detected in endothelial cells within DRG, spinal cord, pancreas, colon, muscles and many immune organs. In addition, gp38+ fibroblastic reticular cells (FRCs), rather than tissue macrophages or other immune cells, were found to express high levels of IL-1R1 in colon and many immune organs. A functional test of spleen FRCs showed that they responded rapidly to systemic IL-1ß stimulation in vivo. Taken together, this study provides a rigorous re-examination of IL-1R1 expression in peripheral tissues and reveals tissue FRCs as a previously unappreciated novel high IL-1R1-expressing cell type in peripheral IL-1 signaling.


Assuntos
Estruturas Animais/química , Estruturas Animais/fisiologia , Perfilação da Expressão Gênica , Receptores de Interleucina-1/biossíntese , Animais , Camundongos , Receptores de Interleucina-1/análise , Receptores de Interleucina-1/genética
16.
Sci Rep ; 8(1): 1601, 2018 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-29371632

RESUMO

L-ergothioneine (ET) is a diet-derived amino acid that accumulates at high concentrations in animals and humans. Numerous studies have highlighted its antioxidant abilities in vitro, and possible cytoprotective capabilities in vivo. We investigated the uptake and distribution of ET in various organs by a highly sensitive and specific liquid chromatography coupled tandem mass spectrometry (LC-MS/MS) technique, both before and after oral administration of pure ET (35 and 70 mg/kg/day for 1, 7, and 28 days) to male C57BL6J mice. ET primarily concentrates in the liver and whole blood, and also in spleen, kidney, lung, heart, intestines, eye, and brain tissues. Strong correlations were found between ET and its putative metabolites - hercynine, ET-sulfonate (ET-SO3H), and S-methyl ET. Hercynine accumulates in the brain after prolonged ET administration. This study demonstrates the uptake and distribution of ET and provides a foundation for future studies with ET to target oxidative damage in a range of tissues in human diseases.


Assuntos
Estruturas Animais/química , Antioxidantes/farmacocinética , Ergotioneína/farmacocinética , Administração Oral , Animais , Antioxidantes/administração & dosagem , Cromatografia Líquida , Ergotioneína/administração & dosagem , Masculino , Camundongos Endogâmicos C57BL , Espectrometria de Massas em Tandem
17.
PLoS One ; 12(12): e0189920, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29261770

RESUMO

Wound healing is a complex biological process, and current research finds that jellyfish have a great capacity for promoting growth and healing. However, the underlying mechanisms remain unclear. Thus, this study was conducted to investigate the molecular mechanisms and effects of a tentacle extract (TE) from the jellyfish Cyanea capillata (C. capillata) on cell proliferation and migration in human umbilical vein endothelial cells (HUVECs). First, our results showed that TE at the concentration of 1 µg/ml could promote cell proliferation over various durations, induce a transition of the cells from the G1-phase to the S/G2-phase of the cell cycle, and increase the expression of cell cycle proteins (CyclinB1 and CyclinD1). Second, we found that TE could activate the PI3K/Akt, ERK1/2 and JNK MAPK signaling pathways but not the NF-κB signaling pathway or the apoptosis signaling cascade. Finally, we demonstrated that the TE-induced expression of cell cycle proteins was decreased by ERK1/2 inhibitor PD98059 but not by PI3K inhibitor LY294002 or JNK inhibitor SP600125. Similarly, the TE-enhanced migration ability of HUVECs was also markedly attenuated by PD98059. Taken together, our findings indicate that TE-induced proliferation and migration in HUVECs mainly occurred through the ERK1/2 MAPK signaling pathway. These results are instructively important for further research on the isolation and purification of growth-promoting factors from C. capillata and are hopeful as a means to improve human wound repair in unfavorable conditions.


Assuntos
Estruturas Animais/química , Movimento Celular/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/citologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Cifozoários/anatomia & histologia , Extratos de Tecidos/farmacologia , Animais , Ciclo Celular/efeitos dos fármacos , Proteínas de Ciclo Celular/metabolismo , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Ciclinas/metabolismo , Imunofluorescência , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , NF-kappa B/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação/efeitos dos fármacos , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Cicatrização/efeitos dos fármacos
18.
Cold Spring Harb Protoc ; 2017(12): pdb.prot098566, 2017 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-29196600

RESUMO

This protocol details protein extraction from mouse tissues for immunoprecipitation purposes and has been applied for the performance of large-scale immunoprecipitations of target proteins from various tissues for the identification of associated proteins by mass spectroscopy. The key factors in performing a successful immunoprecipitation directly relate to the abundance of target protein in a particular tissue type and whether or not the embryonic, newborn, or adult mouse-derived tissues contain fibrous and other insoluble material. Several tissue types, including lung and liver as well as carcinomas, contain significant amounts of fibrous tissue that can interfere with an immunoprecipitation.


Assuntos
Estruturas Animais/química , Estruturas Animais/efeitos dos fármacos , Misturas Complexas/isolamento & purificação , Detergentes/metabolismo , Imunoprecipitação/métodos , Proteínas/isolamento & purificação , Animais , Camundongos
19.
Sci Rep ; 7(1): 15162, 2017 11 09.
Artigo em Inglês | MEDLINE | ID: mdl-29123198

RESUMO

As less consumed animal by-product, beef and pork offal have chances to sneak into the authentic ground beef meat products, and thus a rapid and accurate detection and quantification technique is highly required. In this study, Fourier transformed-infrared (FT-IR) spectroscopy was investigated to develop an optimized protocol for analyzing ground beef meat potentially adulterated with six types of beef and pork offal. Various chemometric models for classification and quantification were constructed for the collected FT-IR spectra. Applying optimized chemometric models, FT-IR spectroscopy could differentiate authentic beef meat from adulterated samples with >99% accuracy, to identify the type of offal in the sample with >80% confidence, and to quantify five types of offal in an accurate manner (R 2 > 0.81). An optimized protocol was developed to authenticate ground beef meat as well as identify and quantify the offal adulterants using FT-IR spectroscopy coupled with chemometric models. This protocol offers a limit of detection <10% w/w of offal in ground beef meat and can be applied by governmental laboratories and food industry to rapidly monitor the integrity of ground beef meat products.


Assuntos
Estruturas Animais/química , Análise de Alimentos/métodos , Produtos da Carne , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Animais , Bovinos , Contaminação de Alimentos , Sensibilidade e Especificidade
20.
Sci Rep ; 7(1): 7641, 2017 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-28794497

RESUMO

Tiger sharks were sampled off the western (Ningaloo Reef, Shark Bay) and eastern (the Great Barrier Reef; GBR, Queensland and New South Wales; NSW) coastlines of Australia. Multiple tissues were collected from each shark to investigate the effects of location, size and sex of sharks on δ13C and δ15N stable isotopes among these locations. Isotopic composition of sharks sampled in reef and seagrass habitats (Shark Bay, GBR) reflected seagrass-based food-webs, whereas at Ningaloo Reef analysis revealed a dietary transition between pelagic and seagrass food-webs. In temperate habitats off southern Queensland and NSW coasts, shark diets relied on pelagic food-webs. Tiger sharks occupied roles at the top of food-webs at Shark Bay and on the GBR, but not at Ningaloo Reef or off the coast of NSW. Composition of δ13C in tissues was influenced by body size and sex of sharks, in addition to residency and diet stability. This variability in stable isotopic composition of tissues is likely to be a result of adaptive foraging strategies that allow these sharks to exploit multiple shelf and offshore habitats. The trophic role of tiger sharks is therefore both context- and habitat-dependent, consistent with a generalist, opportunistic diet at the population level.


Assuntos
Organismos Aquáticos/fisiologia , Comportamento Alimentar , Cadeia Alimentar , Tubarões/fisiologia , Estruturas Animais/química , Animais , Austrália , Biometria , Isótopos de Carbono/análise , Ecossistema , Feminino , Masculino , Isótopos de Nitrogênio/análise , Comportamento Predatório , Tubarões/anatomia & histologia
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