Detection of Parvalbumin in Various Fish Species by 1D and 2D Electrophoresis and Western Blots Using a Commercially Available Anti-frog Parvalbumin MAb

In plant and animal nematode parasites, proteins derived from esophageal gland cells have been shown to be important in the host-nematodes relationship but little is known about the allergenic potential of these proteins in the genus Anisakis. Taking into account the increase of anisakiasis and allergies related to these nematodes, immunoreactive properties of gland cell proteins were investigated. Two hundred ventricles were manually dissected from L3 stage larvae of Aniskakis simplex s.s. to allow direct protein analysis. Denaturing gel electrophoresis followed by monochromatic silver staining which revealed the presence of differential (enriched) proteins when compared to total nematode extracts. Such comparison was performed by means of 1D and 2D electrophoresis. Pooled antisera from Anisakis spp.-allergic patients were used in western blots revealing the presence of 13 immunoreactive bands in the ventricular extracts in 1D, with 82 spots revealed in 2D. The corresponding protein bands and spots were excised from the silver-stained gel and protein assignation was made by MALDI-TOF/TOF. A total of 13 (including proteoforms) were unambiguously identified. The majority of these proteins are known to be secreted by nematodes into the external environment, of which three are described as being major allergens in other organisms with different phylogenetic origin and one is an Anisakis simplex allergen.

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Characterization and evolution of spearfishing in Madeira archipelago, Eastern Atlantic

Aquatic Living Resources ◽

10.1051/alr/2020015 ◽

2020 ◽

Vol 33 ◽

pp. 15

Author(s):

Roi Martínez-Escauriaza ◽

Claudio Vieira ◽

Lídia Gouveia ◽

Nuno Gouveia ◽

Margarida Hermida

Keyword(s):

Fish Species ◽

Young Men ◽

Coastal Areas ◽

Teleost Fishes ◽

The North ◽

Southeast Coast ◽

Madeira Archipelago ◽

First Time

Data obtained from licenses of spearfishers and surveys conducted in 2004 and 2017 allowed for the analysis, for the first time, of the practice of spearfishing in the Madeira archipelago. Only a small percentage of the population practices spearfishing, mostly local young men. Most of them practice the activity with a partner throughout most of the year and along most of the island's coastal areas, although preferentially along the North and Southeast coast. Results show how, in recent years, despite the population of spearfishers decreasing, the abundance in the annual catch potentially increased, probably due to the higher investment of time in this activity. It has been observed that many fishers complement their catches with manual collecting of invertebrates. Overall, 40 teleost fishes and also 4 crustaceans and 8 molluscs were identified. The most frequently captured fish species were parrotfish and white seabream, while limpets were the most collected invertebrates in both selected periods.

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Functional Characterization of PM6/13, a β3-specific (GPIIIa/CD61) Monoclonal Antibody that Shows Preferential Inhibition of Fibrinogen Binding over Fibronectin Binding to Activated Human Platelets

Thrombosis and Haemostasis ◽

10.1055/s-0037-1614247 ◽

1998 ◽

Vol 79 (01) ◽

pp. 177-185 ◽

Cited By ~ 10

Author(s):

Ashia Siddiqua ◽

Michael Wilkinson ◽

Vijay Kakkar ◽

Yatin Patel ◽

Salman Rahman ◽

...

Keyword(s):

Monoclonal Antibody ◽

Platelet Aggregation ◽

Platelet Rich Plasma ◽

Functional Characterization ◽

Human Platelets ◽

Western Blots ◽

Activated Platelets ◽

Reducing Conditions ◽

Fibronectin Binding

SummaryWe report the characterization of a monoclonal antibody (MAb) PM6/13 which recognises glycoprotein IIIa (GPIIIa) on platelet membranes and in functional studies inhibits platelet aggregation induced by all agonists examined. In platelet-rich plasma, inhibition of aggregation induced by ADP or low concentrations of collagen was accompanied by inhibition of 5-hydroxytryptamine secretion. EC50 values were 10 and 9 [H9262]g/ml antibody against ADP and collagen induced responses respectively. In washed platelets treated with the cyclooxygenase inhibitor, indomethacin, PM6/13 inhibited platelet aggregation induced by thrombin (0.2 U/ml), collagen (10 [H9262]g/ml) and U46619 (3 [H9262]M) with EC50 = 4, 8 and 4 [H9262]g/ml respectively, without affecting [14C]5-hydroxytryptamine secretion or [3H]arachidonate release in appropriately labelled cells. Studies in Fura 2-labelled platelets revealed that elevation of intracellular calcium by ADP, thrombin or U46619 was unaffected by PM6/13 suggesting that the epitope recognised by the antibody did not influence Ca2+ regulation. In agreement with the results from the platelet aggregation studies, PM6/13 was found to potently inhibit binding of 125I-fibrinogen to ADP activated platelets. Binding of this ligand was also inhibited by two other MAbs tested, namely SZ-21 (also to GPIIIa) and PM6/248 (to the GPIIb-IIIa complex). However when tested against binding of 125I-fibronectin to thrombin stimulated platelets, PM6/13 was ineffective in contrast with SZ-21 and PM6/248, that were both potent inhibitors. This suggested that the epitopes recognised by PM6/13 and SZ-21 on GPIIIa were distinct. Studies employing proteolytic dissection of 125I-labelled GPIIIa by trypsin followed by immunoprecipitation with PM6/13 and analysis by SDS-PAGE, revealed the presence of four fragments at 70, 55, 30 and 28 kDa. PM6/13 did not recognize any protein bands on Western blots performed under reducing conditions. However Western blotting analysis with PM6/13 under non-reducing conditions revealed strong detection of the parent GP IIIa molecule, of trypsin treated samples revealed recognition of an 80 kDa fragment at 1 min, faint recognition of a 60 kDa fragment at 60 min and no recognition of any product at 18 h treatment. Under similar conditions, SZ-21 recognized fragments at 80, 75 and 55 kDa with the 55kDa species persisting even after 18 h trypsin treatment. These studies confirm the epitopes recognised by PM6/13 and SZ-21 to be distinct and that PM6/13 represents a useful tool to differentiate the characteristics of fibrinogen and fibronectin binding to the GPIIb-IIIa complex on activated platelets.

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Cross-Linked αsγt-Chain Hybrids in Plasma Clots Studied by 1D- and 2D Electrophoresis and Western Blotting

Thrombosis and Haemostasis ◽

10.1055/s-0038-1649601 ◽

1993 ◽

Vol 70 (03) ◽

pp. 438-442 ◽

Cited By ~ 6

Author(s):

B Grøn ◽

C Filion-Myklebust ◽

S Bjørnsen ◽

P Haidaris ◽

F Brosstad

Keyword(s):

Molecular Weight ◽

Monoclonal Antibodies ◽

Human Plasma ◽

Western Blotting ◽

Isoelectric Point ◽

Factor Xiii ◽

Cross Linking ◽

2D Electrophoresis ◽

Complex Phenomenon ◽

Fibrinopeptide A

SummaryFibrinogen and fibrin related chains in reduced human plasma as well as the bonds interlinking partially cross-linked fibrin from plasma clots have been studied by means of 1D- and 2D electrophoresis and Western blotting. Immunovisualization of reduced plasma or partially cross-linked fibrin with monoclonal antibodies specific for the α-chains or the γ-chains have shown that several bands represent material belonging to both chains. In order to decide whether these bands constitute αγ-chain hybrids or superimposed α- and γ-chain dimers, the cross-linked material was separated according to both isoelectric point (pI) and molecular weight (MW) using Pharmacia’s Multiphor II system. Western blotting of the second dimension gels revealed that partially cross-linked fibrin contains αsγt-chain hybrids and γ- polymers, in addition to the well-known γ-dimers and α-polymers. The main αsγt-chain hybrid has a pI between that of the α- and the γ-chains, a MW of about 200 kDa and contains Aα-chains with intact fibrinopeptide A (FPA). It was also observed that soluble fibrinogen/fibrin complexes as well as partially cross-linked fibrin contain degraded α-dimers with MWs close to the γ-dimers. These findings demonstrate that factor XIII-catalyzed cross-linking of fibrin is a more complex phenomenon than earlier recognized.

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DISTRIBUTION, BIOLOGY AND POPULATION STATUS OF BENTO-PELAGIC FISH SPECIES IN THE AREA OF THE SEA CHANNEL OF CARGO TRANSPORTATION IN THE NORTH-EAST CASPIAN

Biological journal ◽

10.32743/2658-6460.2020.5.16.294 ◽

2020 ◽

Vol 16 (5) ◽

Author(s):

Esbol Kasymbekov ◽

Berdibek Abilov

Keyword(s):

Fish Species ◽

Pelagic Fish ◽

Population Status ◽

North East ◽

The North ◽

Cargo Transportation

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Effects of Drying Methods on the Biochemical Composition and Organoleptic Properties of Two Freshwater Fish Species (Mormyrus rume and Labeo coubie) of Cross River Obubra, Nigeria

Journal of Marine Science Research and Oceanography ◽

10.33140/jmsro.02.01.01 ◽

2019 ◽

Vol 2 (1) ◽

Keyword(s):

Moisture Content ◽

Fish Species ◽

Nutrition Status ◽

Drying Methods ◽

Organoleptic Properties ◽

Nutrient Quality ◽

Oven Drying ◽

Fish Samples ◽

Fish And Fishery Products ◽

Mormyrus Rume

Consumers all over the world are increasingly becoming aware of the health and nutrition status of fish and fishery products. There have develop some preference for fish species and even their processing methods. This study aims to investigate the effects of two drying methods (smoking and oven drying) on the biochemical components and organoleptic properties of two less preferred food fishes abundant in the study area. The fish samples Mormyrus rume and Labeo coubie were purchased from fresh landings of fishermen at Ahaha beach. The biochemical assay of the moisture, protein, fat, carbohydrate, ash, calcium (Ca), magnesium (Mg), phosphorus (P), iron (Fe), sodium (Na) and potassium (K) were conducted in the labouratory using standard methods. The proximate composition of raw M. rume was determined as moisture 70.38%, protein 17.43%, carbohydrate 1.13%, fat 5.93% and ash 2.77% while oven dried were 18.78, 63.85, 3.37, 8.73 and 7.58% respectively. All the proximate parameters and gross energy level investigated were higher (p < 0.05) in M. rume except the moisture content of dried samples of L. coubie. The mineral content were in the order Ca < Mg < Fe < Na < P and < K in both species and were higher in M. rume except Na. The drying methods showed increase in the proximate and mineral compositions in the order raw < smoking and < oven drying except moisture content that decreased respectively is both species. Organoleptic properties revealed that dried samples (smoked and oven) were not significantly different (p < 0.05) in taste, flavour, texture and overall acceptance, however while oven dried had better colour (4.33), smoked dried taste (4.63) better. The drying methods shows concentration of the required nutrients in human diets and were also found to be most palatable by the panellist. To improve fish nutrient quality smoke and oven drying should be encourage.

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