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1.
Fish Shellfish Immunol ; 127: 965-974, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35843528

RESUMO

The aim of this study was to investigate the seasonal changes in the epidermal structure and the innate immunity parameters of skin mucus in rainbow trout. The skin epidermis and mucus samples were collected over three consecutive seasons including winter, spring and late summer from three different weight groups i.e., 2-20 g (W1), 100-200 g (W2) and 400-600 g (W3) fish. The skin mucosal immunity analysis of rainbow trout showed that the haemagglutination activity increased significantly with increasing fish size from W1 to W3 in all three seasons, while no significant seasonal changes occurred in haemagglutination activity. Moreover, the bactericidal activity against fish pathogens increased significantly with increasing water bacterial load in late summer. The SDS-PAGE analysis of mucus showed a high amount of low molecular weight proteins (<35 kDa) in the late summer that was correlated with the increase in bactericidal activity. Histological analysis of the epidermis structure of rainbow trout skin showed that the density and size of goblet cells and consequently the mucus secretion significantly increased in W3 group in all seasons. In all three weight groups of fish, the density of goblet cells significantly increased from winter to spring and late summer along with increasing water temperature. Moreover, the goblet cell density showed a significant positive relationship with the soluble protein concentration and haemagglutination activity (p < 0.01). The results of this study demonstrated the more active immune role of the skin epidermal cells and mucus in rainbow trout during summer to protect fish against the pathogenic microorganisms. Given its potent bactericidal properties and the lack of haemolytic activity, the rainbow trout mucus might be used as a safe and inexpensive source for developing antimicrobial agents to prevent and treat some bacterial diseases in human and fish.


Assuntos
Doenças dos Peixes , Oncorhynchus mykiss , Agricultura , Animais , Epiderme , Humanos , Estações do Ano , Pele , Água/análise
2.
Proc Natl Acad Sci U S A ; 115(38): 9456-9461, 2018 09 18.
Artigo em Inglês | MEDLINE | ID: mdl-30190433

RESUMO

With quantum computers of significant size now on the horizon, we should understand how to best exploit their initially limited abilities. To this end, we aim to identify a practical problem that is beyond the reach of current classical computers, but that requires the fewest resources for a quantum computer. We consider quantum simulation of spin systems, which could be applied to understand condensed matter phenomena. We synthesize explicit circuits for three leading quantum simulation algorithms, using diverse techniques to tighten error bounds and optimize circuit implementations. Quantum signal processing appears to be preferred among algorithms with rigorous performance guarantees, whereas higher-order product formulas prevail if empirical error estimates suffice. Our circuits are orders of magnitude smaller than those for the simplest classically infeasible instances of factoring and quantum chemistry, bringing practical quantum computation closer to reality.

3.
Rapid Commun Mass Spectrom ; 31(10): 842-850, 2017 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-28295771

RESUMO

RATIONALE: 17ß-Estradiol (E2), estrone (E1) and estriol (E3) are steroid hormones responsible for the regulation of the female reproductive system. Estradiol is planned to be used to feminize eels in aquaculture in order to improve their size and marketability. The residual levels of these hormones in fish tissue must be monitored to meet the requirements of food regulatory agencies. Few studies have studied these hormones in complex biological matrices such as fish tissue. METHODS: We developed a method to analyze E1, E2 and E3 in fish tissue using liquid chromatography in combination with differential ion mobility spectrometry (DMS) and tandem mass spectrometry (MS/MS). The mass spectrometer was operated in negative polarity selected reaction monitoring (SRM) mode. To test the performance of this method, residual levels of E1, E2 and E3 were measured in the muscle tissue of juvenile eels subjected to feminization treatment with E2. RESULTS: We report that following 17ß-estradiol treatment, E2 is rapidly metabolized from the eel tissue, with a 50% depletion rate per day. Five days post-treatment, E2 returned to the level found in non-treated controls, similar to levels found in wild mature female eels. CONCLUSIONS: The method presented herein allows the quantitative analysis of E1, E2 and E3 in fish tissue samples. Under the experimental conditions, E2 in fish tissue samples returned to physiological levels post hormonal treatment. Copyright © 2017 John Wiley & Sons, Ltd.


Assuntos
Cromatografia Líquida/métodos , Resíduos de Drogas/análise , Estradiol/análise , Estriol/análise , Estrona/análise , Anguilla , Animais , Feminino , Produtos Pesqueiros/análise , Limite de Detecção , Músculo Esquelético/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem
4.
Artigo em Inglês | MEDLINE | ID: mdl-28089856

RESUMO

This research investigated how ploidy level (diploid versus triploid) affects the heat shock protein (HSP) response in erythrocytes under different thermal stress regimes, both in vivo and in vitro, in Atlantic salmon (Salmo salar) and brook charr (Salvelinus fontinalis) in order to address the question of why triploids typically have reduced thermal tolerance. A preliminary study confirmed that identical volumes of diploid and triploid erythrocytes (which equates to a smaller number of larger cells for triploids compared to diploids) did not differ in total protein synthesis rates. After chronic (100d) acclimation of fish to 5, 15 and 25°C, triploid erythrocytes had lower HSP70, HSP90, heat shock factor 1 (HSF1) and ubiquitin (free and total) levels than diploids in both species. Furthermore, Atlantic salmon erythrocytes showed significantly higher protein breakdown (based on conjugated ubiquitin levels) in triploids than diploids after acute heat stress in vitro, but no significant difference was detected between ploidies after acute cold stress. These results indicate that: 1) triploid erythrocytes synthesize more total protein per cell than diploids as a result of increased cell size; 2) triploids have sufficient total HSP levels for survival under low stress conditions; and 3) the lower basal titres of HSPs in triploids may be a handicap when combating acute stress. Taken together, this suggests that triploids are limited in their ability to withstand thermal stress because of a reduced ability to maintain proteostasis under stressful conditions.


Assuntos
Aclimatação , Diploide , Eritrócitos/metabolismo , Proteínas de Choque Térmico/biossíntese , Salmão/fisiologia , Triploidia , Truta/fisiologia , Animais , Aquicultura , Tamanho Celular , Temperatura Baixa/efeitos adversos , Proteínas de Ligação a DNA/biossíntese , Proteínas de Ligação a DNA/sangue , Proteínas de Ligação a DNA/metabolismo , Eritrócitos/citologia , Proteínas de Peixes/biossíntese , Proteínas de Peixes/sangue , Proteínas de Peixes/metabolismo , Regulação da Expressão Gênica , Proteínas de Choque Térmico HSP70/biossíntese , Proteínas de Choque Térmico HSP70/sangue , Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Choque Térmico HSP90/biossíntese , Proteínas de Choque Térmico HSP90/sangue , Proteínas de Choque Térmico HSP90/metabolismo , Fatores de Transcrição de Choque Térmico , Proteínas de Choque Térmico/sangue , Proteínas de Choque Térmico/metabolismo , Técnicas In Vitro/veterinária , Masculino , Estabilidade Proteica , Salmão/genética , Salmão/metabolismo , Especificidade da Espécie , Estresse Fisiológico , Fatores de Transcrição/biossíntese , Fatores de Transcrição/sangue , Fatores de Transcrição/metabolismo , Truta/genética , Truta/metabolismo , Ubiquitina/biossíntese , Ubiquitina/sangue , Ubiquitina/metabolismo
5.
Heart Lung Circ ; 26(12): 1297-1302, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28318819

RESUMO

BACKGROUND: The Magnetic Navigation System (MNS) catheter was shown to be stable in the presence of significant cardiac wall motion and delivered more effective lesions compared to manual control. This stability could potentially make AV junctional re-entrant tachycardia (AVNRT) ablation safer. The aim of this study is to describe the method of mapping and ablation of AVNRT with MNS and 3-D electro-anatomical mapping system (CARTO, Biosense Webster, Diamond bar, CA, USA) anatomical mapping, with a view to improve the safety of ablation. METHODS: The method of precise mapping and ablation with MNS is described. Consecutive AVNRT cases (n=30) from 2012 January to 2015 November, in which magnetic navigation was used, are analysed. RESULTS: Ablation was successful in 27 (90%) out of 30 patients. In three cases, ablation was abandoned due to the proximity of the three-dimensional His image to the potential ablation site. No complications, including AV nodal injury, occurred. The distance from the nearest His position to successful ablation site in both LAO and RAO projections of CARTO images was 26.4±8.8 and 27±7.7mm respectively. Only in two (9%) patients, ablation needed to be extended superior to the plane of coronary sinus ostium, towards the His bundle region, to achieve slow pathway modification. CONCLUSION: AVNRT ablation with MNS allows for accurate mapping of the AV node and stable ablation at a safe distance, which could help avoid AV nodal injury. We recommend this modality for younger patients with AVNRT.


Assuntos
Ablação por Cateter/métodos , Magnetismo/métodos , Cirurgia Assistida por Computador/métodos , Adulto , Seio Coronário , Eletrocardiografia , Feminino , Seguimentos , Humanos , Masculino , Estudos Retrospectivos , Taquicardia por Reentrada no Nó Atrioventricular/diagnóstico , Taquicardia por Reentrada no Nó Atrioventricular/fisiopatologia , Taquicardia por Reentrada no Nó Atrioventricular/cirurgia , Resultado do Tratamento
6.
Philos Trans A Math Phys Eng Sci ; 374(2059)2016 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-26667914

RESUMO

It is now well documented that over 400 subglacial lakes exist across the bed of the Antarctic Ice Sheet. They comprise a variety of sizes and volumes (from the approx. 250 km long Lake Vostok to bodies of water less than 1 km in length), relate to a number of discrete topographic settings (from those contained within valleys to lakes that reside in broad flat terrain) and exhibit a range of dynamic behaviours (from 'active' lakes that periodically outburst some or all of their water to those isolated hydrologically for millions of years). Here we critique recent advances in our understanding of subglacial lakes, in particular since the last inventory in 2012. We show that within 3 years our knowledge of the hydrological processes at the ice-sheet base has advanced considerably. We describe evidence for further 'active' subglacial lakes, based on satellite observation of ice-surface changes, and discuss why detection of many 'active' lakes is not resolved in traditional radio-echo sounding methods. We go on to review evidence for large-scale subglacial water flow in Antarctica, including the discovery of ancient channels developed by former hydrological processes. We end by predicting areas where future discoveries may be possible, including the detection, measurement and significance of groundwater (i.e. water held beneath the ice-bed interface).

7.
Philos Trans A Math Phys Eng Sci ; 374(2059)2016 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-26667913

RESUMO

Accessing and sampling subglacial environments deep beneath the Antarctic Ice Sheet presents several challenges to existing drilling technologies. With over half of the ice sheet believed to be resting on a wet bed, drilling down to this environment must conform to international agreements on environmental stewardship and protection, making clean hot-water drilling the most viable option. Such a drill, and its water recovery system, must be capable of accessing significantly greater ice depths than previous hot-water drills, and remain fully operational after connecting with the basal hydrological system. The Subglacial Lake Ellsworth (SLE) project developed a comprehensive plan for deep (greater than 3000 m) subglacial lake research, involving the design and development of a clean deep-ice hot-water drill. However, during fieldwork in December 2012 drilling was halted after a succession of equipment issues culminated in a failure to link with a subsurface cavity and abandonment of the access holes. The lessons learned from this experience are presented here. Combining knowledge gained from these lessons with experience from other hot-water drilling programmes, and recent field testing, we describe the most viable technical options and operational procedures for future clean entry into SLE and other deep subglacial access targets.

8.
J Sports Sci ; 31(7): 787-94, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23521618

RESUMO

The purpose of this study was to examine the distribution of pace self-selected by cyclists of varying ability, biological age and sex performing in a mountain bike World Championship event. Data were collected on cyclists performing in the Elite Male (ELITEmale; n = 75), Elite Female (ELITEfemale; n = 50), Under 23 Male (U23male; n = 62), Under 23 Female (U23female; n = 34), Junior Male (JNRmale; n = 71) and Junior Female (JNRfemale; n = 30) categories of the 2009 UCI Cross-Country Mountain Bike World Championships. Split times were recorded for the top, middle and bottom 20% of all finishers of each category. Timing splits were positioned to separate the course into technical and non-technical, uphill, downhill and rolling/flat sections. Compared with bottom performers, top performers in all male categories (ELITEmale, U23male, JNRmale) maintained a more even pace over the event as evidenced by a significantly lower standard deviation and range in average lap speed. Top performers, males, and ELITEmale athletes spent a lower percentage of overall race time on technical uphill sections of the course, compared with middle and bottom placed finishers, females, and JNRmale athletes, respectively. Better male performers adopt a more even distribution of pace throughout cross-country mountain events. Performance of lower placed finishers, females and JNRmale athletes may be improved by enhancing technical uphill cycling ability.


Assuntos
Desempenho Atlético , Ciclismo , Comportamento Competitivo , Resistência Física , Esforço Físico , Adolescente , Adulto , Feminino , Humanos , Masculino , Fatores Sexuais , Adulto Jovem
9.
Nat Commun ; 14(1): 6507, 2023 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-37875503

RESUMO

The East Antarctic Ice Sheet (EAIS) has its origins ca. 34 million years ago. Since then, the impact of climate change and past fluctuations in the EAIS margin has been reflected in periods of extensive vs. restricted ice cover and the modification of much of the Antarctic landscape. Resolving processes of landscape evolution is therefore critical for establishing ice sheet history, but it is rare to find unmodified landscapes that record past ice conditions. Here, we discover an extensive relic pre-glacial landscape preserved beneath the central EAIS despite millions of years of ice cover. The landscape was formed by rivers prior to ice sheet build-up but later modified by local glaciation before being dissected by outlet glaciers at the margin of a restricted ice sheet. Preservation of the relic surfaces indicates an absence of significant warm-based ice throughout their history, suggesting any transitions between restricted and expanded ice were rapid.

10.
JACC Clin Electrophysiol ; 7(10): 1274-1284, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34454889

RESUMO

OBJECTIVES: This study describes the clinical and electrophysiological characteristics of basal-septal ventricular tachycardias (VTs) in patients with structural heart disease (SHD). BACKGROUND: The basal septum is a common source of VT in patients with SHD. METHODS: Data from 312 consecutive patients with SHD undergoing catheter ablation of ventricular arrhythmias were reviewed. RESULTS: Thirty-three basal-septal VTs in 31 patients (mean age 67.4 ± 14.2 years, mean left ventricular ejection fraction [LVEF] 42% ± 15%) were identified. Patients with VTs with left ventricular basal-septal breakthrough were more likely to have ischemic cardiomyopathy and lower LVEF; patients with right ventricular basal-septal VT were more likely to have sarcoidosis or right ventricular cardiomyopathy of unknown significance, with higher LVEF. Atrioventricular block was present in 45% of patients and intraventricular block including persistent biventricular pacing in 77%. Unipolar scar was larger than bipolar scar (area 18.8% ± 19.4% vs 12.7% ± 14.6%; P < 0.001). VTs with right bundle branch block configuration and S wave in lead V6 with positive V3/V4 polarity consistently indicated left ventricular basal-septal breakthrough. Inferior limb-lead discordance with right bundle branch block configuration and "reverse pattern break in lead V2" were identified in left ventricular basal inferior-septal origin in 3 patients. VT noninducibility was achieved in 55%, and VT recurred in 42% of patients after a single procedure, but VT burden was significantly reduced after ablation (59 episodes before vs 2 episodes after ablation; P = 0.02). CONCLUSIONS: Basal-septal VTs in patients with SHD have a distinct clinical, electrocardiographic, and electrophysiological profile depending on the breakthrough site, accompanied by a deep intramural septal substrate that limits procedural success after catheter ablation.


Assuntos
Cardiomiopatias , Ablação por Cateter , Taquicardia Ventricular , Idoso , Idoso de 80 Anos ou mais , Cardiomiopatias/cirurgia , Humanos , Pessoa de Meia-Idade , Volume Sistólico , Taquicardia Ventricular/cirurgia , Função Ventricular Esquerda
11.
J Innov Card Rhythm Manag ; 12(4): 4467-4472, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33936862

RESUMO

In ischemic cardiomyopathy, endocardial reentry has traditionally been the mechanistic paradigm for understanding ventricular tachycardia (VT). However, recognition is growing that epicardial myocardium is a critical component for VT substrate, even in patients with ischemic cardiomyopathy. In this report, we present a novel case of a three-dimensional VT reentry involving epicardial components and an endocardial exit.

12.
Chemosphere ; 263: 128315, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33297250

RESUMO

The traditional approach to extracting estrogens from water matrices, solid-phase extraction (SPE), presents a number of challenges when applied to complex wastewater matrices. Conversely, the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) clean-up method offers an alternative sample preparation approach that omits sample filtration and overcomes additional challenges associated with SPE. The objective of this study was to implement and validate a scaled QuEChERS method, using a standard addition approach, for extracting estrone (E1), 17ß-estradiol (E2), and estriol (E3) from the estrogenic influent of a recirculating aquaculture system containing American eels (Anguilla rostrata). While traditional QuEChERS protocols do not facilitate considerable sample concentration, a 500-fold concentration factor was implemented for reliable quantitation of parts-per-trillion concentrations of estrogens from an initial sample volume of 20 mL to a final extract volume of 40 µL. Following analysis via high-performance liquid chromatography-mass spectrometry, excellent process efficiencies were observed at spiked concentrations of 10 and 50 ng L-1 for E2 and E1 (101 to 111%; %RSD ≤ 16), and moderate to acceptable process efficiencies were achieved for E3 (75 to 87%; %RSD ≤ 16). Validation of method parameters, including specificity, linearity, accuracy (recovery and process efficiencies), precision (intra-day precision, and inter-day precision), matrix effects, method detection limit, and limit of quantitation, led to reliable quantitation of unknown concentrations of E1, E2, and E3 in the aquaculture influent as low as 52, 20, and 33 ng L-1, respectively. This study provides a validated analytical method for waste systems requiring quantitation of estrogens in their complex wastewater matrices.


Assuntos
Estrogênios , Água , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Estrogênios/análise , Limite de Detecção , Extração em Fase Sólida , Espectrometria de Massas em Tandem
13.
Artigo em Inglês | MEDLINE | ID: mdl-18342561

RESUMO

The mucus layer on the surface of fish consists of several antimicrobial agents that provide a first line of defense against invading pathogens. To date, little is known about the antimicrobial properties of the mucus of Arctic char (Salvelinus alpinus), brook trout (S. fontinalis), koi carp (Cyprinus carpio sub sp. koi), striped bass (Morone saxatilis), haddock (Melanogrammus aeglefinus) and hagfish (Myxine glutinosa). The epidermal mucus samples from these fish were extracted with acidic, organic and aqueous solvents to identify potential antimicrobial agents including basic peptides, secondary metabolites, aqueous and acid soluble compounds. Initial screening of the mucus extracts against a susceptible strain of Salmonella enterica C610, showed a significant variation in antimicrobial activity among the fish species examined. The acidic mucus extracts of brook trout, haddock and hagfish exhibited bactericidal activity. The organic mucus extracts of brook trout, striped bass and koi carp showed bacteriostatic activity. There was no detectable activity in the aqueous mucus extracts. Further investigations of the activity of the acidic mucus extracts of brook trout, haddock and hagfish showed that these fish species had specific activity for fish and human pathogens, demonstrating the role of fish mucus in antimicrobial protection. In comparison to brook trout and haddock, the minimum bactericidal concentrations of hagfish acidic mucus extracts were found to be approximately 1.5 to 3.0 times lower against fish pathogens and approximately 1.6 to 6.6 folds lower for human pathogens. This preliminary information suggests that the mucus from these fish species may be a source of novel antimicrobial agents for fish and human health related applications.


Assuntos
Anti-Infecciosos/farmacologia , Epiderme/química , Peixes/metabolismo , Muco/química , Extratos de Tecidos/farmacologia , Animais , Bactérias/efeitos dos fármacos , Eletroforese em Gel de Poliacrilamida , Epiderme/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Muco/efeitos dos fármacos , Leveduras/efeitos dos fármacos
14.
Nat Commun ; 9(1): 4576, 2018 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-30385741

RESUMO

Satellite imagery reveals flowstripes on Foundation Ice Stream parallel to ice flow, and meandering features on the ice-shelf that cross-cut ice flow and are thought to be formed by water exiting a well-organised subglacial system. Here, ice-penetrating radar data show flow-parallel hard-bed landforms beneath the grounded ice, and channels incised upwards into the ice shelf beneath meandering surface channels. As the ice transitions to flotation, the ice shelf incorporates a corrugation resulting from the landforms. Radar reveals the presence of subglacial water alongside the landforms, indicating a well-organised drainage system in which water exits the ice sheet as a point source, mixes with cavity water and incises upwards into a corrugation peak, accentuating the corrugation downstream. Hard-bedded landforms influence both subglacial hydrology and ice-shelf structure and, as they are known to be widespread on formerly glaciated terrain, their influence on the ice-sheet-shelf transition could be more widespread than thought previously.

15.
Comp Biochem Physiol B Biochem Mol Biol ; 148(3): 256-63, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17618153

RESUMO

Fish epidermal mucus and its components provide the first line of defense against pathogens. Little is known about the role of epidermal mucus enzymes in the innate immune system of fish species such as Arctic char (Salvelinus alpinus), brook trout (S. fontinalis), koi carp(Cyprinus carpio), striped bass (Morone saxatilis), haddock, (Melanogrammus aeglefinus), Atlantic cod (Gadus morhua) and hagfish (Myxine glutinosa). The epidermal mucus samples from these fish were analysed for the specific activities of various hydrolytic enzymes including lysozyme, alkaline phosphatase, cathepsin B and proteases and the enzyme levels were compared among the fish species. Of all the species hagfish mucus showed a high activity for lysozyme and proteases and koi carp mucus had the highest levels of alkaline phosphatase and cathepsin B. A wide variation in enzyme activities was observed among the seven species and also between species of same family such as Arctic char and brook trout (salmonidae), haddock and cod (gadidae). Only lysozyme levels showed a marked variation with salinity where seawater fish showed approximately two times higher lysozyme activity than freshwater-reared fish species. Characterization of proteases with specific inhibitors showed Arctic char, brook trout, haddock and cod having higher levels of serine over metalloproteases whereas koi carp and striped bass had higher levels of metalloproteases over serine proteases. In contrast, hagfish had almost equal proportion of both serine and metalloproteases. This study demonstrates variation in the level of hydrolytic enzymes in the epidermal mucus of fish. These results provide preliminary information for a better understanding of the role of epidermal mucus and its components in the fish innate immune system.


Assuntos
Epiderme/imunologia , Imunidade Inata , Muco/imunologia , Animais , Peixes , Especificidade da Espécie
16.
Comp Biochem Physiol B Biochem Mol Biol ; 144(4): 488-95, 2006 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16781177

RESUMO

A study was conducted to compare astaxanthin binding ability of solubilized muscle proteins of Atlantic salmon (Salmo salar L.), haddock (Melanogrammus aeglefinus L.) and Atlantic halibut (Hippoglossus hippoglossus L.). Muscle proteins of juvenile Atlantic salmon, haddock and halibut were solubilized by sequential extraction of muscle tissue using low ionic strength solutions. Electrophoretic protein profiles of the six solubilized fractions from these species were similar. Each solubilized fraction from the three species was examined for its relative astaxanthin binding capacity. The amount of bound astaxanthin was significantly different (P<0.05) among the six fractions of each species. Significant differences in astaxanthin binding were only found for fractions A and E among the species. The amount of bound astaxanthin in various fractions of each species showed a good correlation (R2=0.80-0.92) with the ANS (8-anilino-1-naphthalenesulfonate) fluorescence intensity of those fractions. The pattern and extent of astaxanthin binding to the muscle proteins of juvenile salmon, haddock and halibut is comparable to that reported previously for adult Atlantic salmon (Saha, M.R., Ross, N.W., Gill, T.A., Olsen, R.E., Lall, S.P., 2005. Development of a method to assess binding of astaxanthin to Atlantic salmon S. salar L. muscle proteins. Aquacult. Res. 36, 336-343.). These combined observations suggest that the carotenoid binding capacity of the muscle proteins of salmon is not the limiting factor in the deposition of carotenoid in their flesh.


Assuntos
Proteínas de Peixes/química , Linguado/metabolismo , Gadiformes/metabolismo , Proteínas Musculares/química , Salmo salar/metabolismo , Animais , Músculos/metabolismo , Ligação Proteica , Xantofilas/química
17.
Comp Biochem Physiol B Biochem Mol Biol ; 144(2): 206-14, 2006 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-16644255

RESUMO

The rubicund pigmentation in salmon and trout flesh is unique and is due to the deposition of dietary carotenoids, astaxanthin and canthaxanthin in the muscle. The present study was undertaken to determine which protein was responsible for pigment binding. Salmon muscle proteins were solubilized by sequential extractions with non-denaturing, low ionic strength aqueous solutions and segregated as such into six different fractions. Approximately 91% of the salmon myofibrillar proteins were solubilized under non-denaturing conditions using a protocol modified from a method described by Krishnamurthy et al. [Krishnamurthy, G., Chang, H.S., Hultin, H.O., Feng, Y., Srinivasan, S., Kelleher. S.D., 1996. Solubility of chicken breast muscle proteins in solutions of low ionic strength. J. Agric. Food Chem. 44: 408-415.] for the dissolution of avian muscle. To our knowledge, this is the first time this solubilization approach has been applied to the study of molecular interactions in myofibrillar proteins. Astaxanthin binding in each fraction was determined using an in vitro binding assay. In addition, SDS-PAGE and quantitative densitometry were used to separate and determine the relative amounts of each of the proteins in the six fractions. The results showed that alpha-actinin was the only myofibrillar protein correlating significantly (P<0.05) with astaxanthin binding. Alpha-actinin was positively identified using electrophoretic techniques and confirmed by tandem mass spectroscopy. Purified salmon alpha-actinin bound synthetic astaxanthin in a molar ratio of 1.11:1.00. The study was repeated using halibut alpha-actinin, which was found to have a molar binding ratio of astaxanthin to alpha-actinin of 0.893:1. These results suggest that the difference in pigmentation between white fish and Atlantic salmon is not due to binding capacity in the muscle, but rather differences in the metabolism or transport of pigment.


Assuntos
Pigmentação/fisiologia , Ligação Proteica/fisiologia , Salmo salar/fisiologia , Actinina/isolamento & purificação , Sequência de Aminoácidos , Animais , Eletroforese em Gel de Poliacrilamida , Linguado/fisiologia , Espectrometria de Massas , Dados de Sequência Molecular , Músculos/química , Pigmentos Biológicos/isolamento & purificação , Xantofilas/metabolismo
18.
Dis Aquat Organ ; 68(1): 29-38, 2005 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-16465831

RESUMO

Aeromonas salmonicida subsp. salmonicida is a Gram-negative bacterium that is the etiological agent of furunculosis, a serious infectious disease of salmonids. Aeromonas spp. are ubiquitous waterborne bacteria responsible for a wide spectrum of diseases among aquatic organisms and humans. Bacterial outer membrane proteins (OMPs) play a significant role in virulence as they comprise the outermost surface in contact with host cells and immune defense factors. To identify the major OMPs of A. salmonicida a proteomic analysis was undertaken using a carbonate OMP-enrichment protocol. The enriched OMP-extracts were separated by 2-dimensional electrophoresis (2-DE) and the spots identified using liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS) via an electrospray ionization source. In total, 76 unique proteins were identified from the 125 spots observed on the 2-D gel. The surface layer (S-layer) VapA protein dominated the A. salmonicida OMP 2-D profile, accounting for 60% of the protein on the 2-D gels. Among the other outer membrane proteins identified were at least 10 porins and various receptors involved in nutrient acquisition. Also identified in the carbonate insoluble fraction were phosphoglycerate kinase, enolase and others that lacked classical export sorting signals. The putative association of these proteins with the cell surface might provide new insights concerning the biological and pathogenic roles of these molecules in A. salmonicida infection. This work represents the first systematic attempt to characterize the cell surface of A. salmonicida.


Assuntos
Aeromonas salmonicida/química , Proteínas da Membrana Bacteriana Externa/classificação , Doenças dos Peixes/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , 2,2'-Dipiridil/metabolismo , Aeromonas salmonicida/metabolismo , Proteínas da Membrana Bacteriana Externa/isolamento & purificação , Proteínas da Membrana Bacteriana Externa/metabolismo , Cromatografia Líquida/métodos , Cromatografia Líquida/veterinária , Eletroforese em Gel Bidimensional/métodos , Regulação Bacteriana da Expressão Gênica , Infecções por Bactérias Gram-Negativas/microbiologia , Ferro/metabolismo , Proteômica , Espectrometria de Massas por Ionização por Electrospray/veterinária
19.
Comp Biochem Physiol B Biochem Mol Biol ; 135(1): 95-108, 2003 May.
Artigo em Inglês | MEDLINE | ID: mdl-12781977

RESUMO

Haddock (Melanogrammus aeglefinus) is a gadoid fish species that deposits dietary lipid mainly in the liver. The fatty acid (FA) beta-oxidation activity of various tissues was evaluated in juvenile haddock fed graded levels of lipid. The catabolism of a radiolabelled FA, [1-(14)C]palmitoyl-CoA, through peroxisomal and mitochondrial beta-oxidation was determined in the liver, red and white muscle of juvenile haddock fed 12, 18 and 24% lipid in the diet. There was no significant increase in the mitochondrial or peroxisomal beta-oxidation activity in the tissues tested as the dietary lipid level increased from 12 to 24%. Peroxisomes accounted for 100% of the beta-oxidation observed in the liver, whereas mitochondrial beta-oxidation dominated in the red (91%) and white muscle (97%) of juvenile haddock. Of the tissues tested, red muscle possessed the highest specific activity for beta-oxidation expressed on a per mg protein or per g wet weight basis. However, white muscle, which forms over 50% of the body mass in gadoid fish was the most important tissue in juvenile haddock for overall FA catabolism. The total lipid and FA composition of these tissues were also determined. This study confirmed that the liver was the major lipid storage organ in haddock. The hepatosomatic index (HSI; 10.0-15.2%) and lipid (73.8-79.3% wet wt.) in the liver increased significantly as dietary lipid was increased from 12 to 24% lipid. There was no significant increase in the lipid composition of the white muscle (0.8% wet wt.), red muscle (1.9% wet wt.) or heart (2.5% wet wt.).


Assuntos
Gorduras na Dieta/farmacologia , Ácidos Graxos/metabolismo , Peixes/metabolismo , Metabolismo dos Lipídeos , Ração Animal/análise , Animais , Radioisótopos de Carbono , Ácidos Graxos/análise , Peixes/sangue , Peixes/crescimento & desenvolvimento , Lipídeos/química , Lipoproteínas VLDL/sangue , Mitocôndrias Hepáticas/química , Mitocôndrias Hepáticas/metabolismo , Músculos/química , Músculos/metabolismo , Miocárdio/química , Miocárdio/metabolismo , Especificidade de Órgãos , Oxirredução/efeitos dos fármacos , Palmitoil Coenzima A/farmacologia
20.
Dis Aquat Organ ; 52(1): 57-68, 2002 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-12517006

RESUMO

Physiological, immunological and biochemical parameters of blood and mucus, as well as skin histology, were compared in 3 salmonid species (rainbow trout Oncorhynchus mykiss, Atlantic salmon Salmo salar and coho salmon O. kisutch) following experimental infection with sea lice Lepeophtheirus salmonis. The 3 salmonid species were cohabited in order to standardize initial infection conditions. Lice density was significantly reduced on coho salmon within 7 to 14 d, while lice persisted in higher numbers on rainbow trout and Atlantic salmon. Lice matured more slowly on coho salmon than on the other 2 species, and maturation was slightly slower on rainbow trout than on Atlantic salmon. Head kidney macrophages from infected Atlantic salmon had diminished respiratory burst and phagocytic capacity at 14 and 21 d post-infection (dpi), while infected rainbow trout macrophages had reduced respiratory burst and phagocytic capacities at 21 dpi, compared to controls. The slower development of lice, coupled with delayed suppression of immune parameters, suggests that rainbow trout are slightly more resistant to lice than Atlantic salmon. Infected rainbow trout and Atlantic salmon showed increases in mucus lysozyme activities at 1 dpi, which decreased over the rest of the study. Mucus lysozyme activities of infected rainbow trout, however, remained higher than controls over the entire period. Coho salmon lysozyme activities did not increase in infected fish until 21 dpi. Mucus alkaline phosphatase levels were also higher in infected Atlantic salmon compared to controls at 3 and 21 dpi. Low molecular weight (LMW) proteases increased in infected rainbow trout and Atlantic salmon between 14 and 21 dpi. Histological analysis of the outer epithelium revealed mucus cell hypertrophy in rainbow trout and Atlantic salmon following infection. Plasma cortisol, glucose, electrolyte and protein concentrations and hematocrit all remained within physiological limits for each species, with no differences occurring between infected and control fish. Our results demonstrate that significant differences in mucus biochemistry and numbers of L. salmonis occur between these species.


Assuntos
Copépodes/crescimento & desenvolvimento , Ectoparasitoses/veterinária , Doenças dos Peixes/imunologia , Oncorhynchus kisutch , Oncorhynchus mykiss , Salmo salar , Fosfatase Alcalina/metabolismo , Animais , Copépodes/patogenicidade , Suscetibilidade a Doenças/veterinária , Ectoparasitoses/imunologia , Ectoparasitoses/parasitologia , Doenças dos Peixes/parasitologia , Interações Hospedeiro-Parasita , Muco/citologia , Muco/enzimologia , Muco/imunologia , Muramidase/metabolismo , Fagocitose , Pele/citologia , Pele/enzimologia , Pele/imunologia , Especificidade da Espécie
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