Lipidomics profile and volatile compounds of squids (Illex argentinus, Ommastrephes Bartram and Dosidicus gigas) in the Argentine, North Pacific Ocean, Equator and Peru by UPLC-triple TOF-MS and HS-SPME-GC-O-MS.

Comprehensive lipid and volatile compound analyses were performed with squids collected from four varied geographical locations to discriminate the regional characteristics. A total of 1442 lipid molecules and 110 volatiles were detected in the squid muscle samples. There were significant differences in the lipid profiles between Argentine squid (Illex argentinus, AGT), North Pacific Ocean squid (Ommastrephes Bartram, NPO), Equatorial squid (Dosidicus gigas, EQ), and Peruvian squid (Dosidicus gigas, PR) muscle. Phosphatidylcholines (14.64%), triacylglycerols (12.42%), and ceramides (10.97%) were the main lipid components. The contents of polyunsaturated fatty acid in phospholipids and in glycerolipids were 30.35-52.05% and 18.11-25.15%, respectively. The volatiles in squids exhibited significant regional variation; 1-pentanol and 1-octanol, 2-ethyl-1-hexanol and terpinen-4-ol, 2,7-ethyl-1-hexanol, 3-methy-1-butanol and 2-propyl-1-pentanol were identified as characteristic flavor compounds in AGT, NPO, EQ, and PR, respectively. Sphingomyelin, phosphatidylserine, phosphatidylethanolamine, and ceramide were strongly correlated with volatiles in squid muscle. Our study is a reference for the lipid nutritional value and flavor compounds of squids.

Optimizing Josephson Junction Reproducibility in 30 kV E-Beam Lithography: An Analysis of Backscattered Electron Distribution.

This paper explores methods to enhance the reproducibility of Josephson junctions, which are crucial elements in superconducting quantum technologies, when employing the Dolan technique in 30 kV e-beam processes. The study explores the influence of dose distribution along the bridge area on reproducibility, addressing challenges related to fabrication sensitivity. Experimental methods include e-beam lithography, with electron trajectory simulations shedding light on the behavior of backscattered electrons. Wedescribe the fabrication of various Josephson junction geometries and analyze the correlation between the success rates of different lithography patterns and the simulated distribution of backscattered electrons. Our findings demonstrate a success rate of up to 96.3% for the double-resist 1-step low-energy e-beam lithography process. As a means of implementation strategy, we provide a geometric example that takes advantage of simulated stability regions to administer a controlled, uniform dose across the junction area, introducing novel features to overcome the difficulties associated with fabricating bridge-like structures.

Effects of Heating Treatment on the Physicochemical and Volatile Flavor Properties of Argentinian Shortfin Squid (Illex argentinus).

In this study, the effect of different heating temperatures (80, 90, 100, and 121 °C) on the physicochemical and volatile flavor properties of fried mantles (Argentinian shortfin) was investigated. The squid mantles were soaked in a maltose syrup solution (20% w/v) for 10 s and fried in soybean oil for 10 s (160 °C), vacuum-packed, and processed at different temperatures for 10 min. Then, the squid mantles were subjected to colorimetric analysis, sensory evaluation, free amino acid analysis, and texture profile analysis. In addition, the volatile organic compounds (VOCs) in the squid mantles were analyzed. The results revealed that lower treating temperatures (80 and 90 °C) improved the chromatic and textural properties, along with organoleptic perception. Additionally, the content of amino acid in the squid mantles treated at 121 °C was significantly lower than that of the samples treated at other temperatures (p < 0.05). Headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) was used to detect 41 VOCs, including their monomers and dimers. Among these detected VOCs, the contents of alcohols, ketones, and pyrazines were positively correlated with temperature. However, the content of aldehydes in the squid mantles gradually decreased as the heating temperature increased (p < 0.05). The combined HS-GC-IMS and E-nose results revealed that the lower temperatures (80 and 90 °C) were more suitable for flavor development and practical processing. This study provides valuable information for properly controlling the heating process of squid products, as well as flavor and practical applications for the aquatic industry.

Assessing the safety and efficacy of EVOH Pre-Embolization in head and neck paraganglioma tumors: A comprehensive analysis of current literature.

BACKGROUND: Head and Neck Paragangliomas are characterized by having a rich blood supply. Presurgical embolization with Onyx as a neoadjuvant treatment is not a consensus regarding its efficacy and safety. Our study aimed to answer this matter through a single-arm meta-analysis. METHODS: We systematically reviewed 4 databases. Sixteen studies were described and suitable papers were selected for meta-analysis of estimated intraoperative blood loss (EBL), percentage of tumor devascularization, and complications associated with embolization. RESULTS: The study identified 198 patients with 203 tumors, aged between 8 and 70 years. Commonly reported symptoms included neck mass perception and cranial nerve impairment. Carotid Body Tumors were most prevalent (127, 62.5 %), followed by jugular (48, 23.6 %), or vagal (29, 14.2 %) tumors. Eight studies reported estimated intraoperative blood loss (EBL) averaging 261.89 ml (95 %CI: 128.96 to 394.81 ml). In an analysis of 9 studies, 99 % (95 %CI: 96 to 100 %) achieved 70 % or more devascularization, and 79 % (95 %CI: 58 to 100 %) achieved 90 % or more devascularization. Complications from endovascular procedures were observed in 3 % (95 %CI: 0 to 8 %) of 96 patients across 10 studies, including 4 facial nerve deficits. Eighteen postoperative neurological deficits were reported across 15 articles. CONCLUSION: Despite acknowledged limitations, with refined indications, EVOH, especially Onyx embolization may significantly bolster patient safety, decreasing EBL and easing surgical resection. Further research with larger studies will refine criteria, optimize techniques, and improve patient care and treatment outcomes in the management of head and neck paragangliomas.

Argentine squid (Illex argentinus): A source of mycosporine-like amino acids with antioxidant properties.

Here we report on the occurrence of mycosporine-like amino acids (MAAs) in the Argentine shortfin squid, Illex argentinus, the second fishery resource mostly exploited in the Argentinean continental shelf. The total content of four MAAs was evaluated by reverse-phase-HPLC in different tissues (eyes, skin, liver, and gonads). Also, the antioxidant activity of crude extracts was assessed by in-vitro determinations: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Folin-Ciocalteu, and ferrous ion-chelating capacity assays. The content of MAAs was found to be almost ten times higher in female gonads than in other tissues (11,89 ± 0,56 mg/g dry weight). Extracts from skin, female gonads and eyes, exhibit higher antioxidant activity than the reference compounds ascorbic acid and TROLOX. Overall, Argentine squid waste is a promising potential source of MAAs with antioxidant and UV photoprotective properties, which could bear interest in food, cosmetic and pharmaceutical industries, thus encouraging maximal and sustainable use of fishing resources.

Aprovechamiento alternativo de calamar (Dosidicus gigas) como fuente de proteína para la fortificación de pan tipo baguette / Alternative use of squid (Dosidicus gigas) flour as a protein source to improve the nutritional quality of baguette bread

Introducción. Los cereales tienen aminoácidos limitantes y mezclarlos con harinas de origen marino presentan un mejor equilibrio en éstos mejorando la composición nutricional en productos de panificación, principalmente de proteína y aminoácidos. Objetivo. Desarrollar panes tipo "baguette" sustituyendo parcialente harina de trigo (HT) por harina de calamar (HC) y evaluar su calidad física, química y sensorial. Materiales y métodos. Se utilizó la técnica de masa esponja incorporando 7,5, 10 y 12,5 % de HC. Análisis físicos: tiempo de amasado, peso y fermentación de la masa, volumen específico, peso de los panes (g), % pérdida humedad y color de miga; análisis químicos: humedad, proteína cruda, cenizas, extracto etéreo, fibra cruda, carbohidratos, aminoácidos indispensables y energía bruta y evaluación sensorial: textura, olor, color y sabor. Resultados. Las propiedades físicas no presentaron diferencias (p>0,05), pero sí (p<0,05) para las variables de color en todos los panes, al igual que en los análisis químicos y perfil de aminoácidos (p<0,05), incrementándose la proteína (15,63 % pan con 7,5 % de HC hasta 23,27 % pan con 12,5 % de HC) y mismo comportamiento de aminoácidos indispensables conforme aumentó la inclusión de HC. Los resultados de la evaluación sensorial fueron similares (p>0,05) para 7,5 % de HC (me gusta), para 10,0 y 12,5 % fue indiferente. Conclusiones. La inclusión de HC (7,5 %) en baguette logró un aumento de proteínas y aminoácidos esenciales mejorando la calidad nutricional, con buena aceptación de acuerdo con la prueba de carácter exploratorio con consumidores que se empleó(AU)

Introduction. Cereals have limiting amino acids and mixing them with flours of marine origin could provide a better balance in these improving the nutritional composition in bakery products, mainly protein and amino acids. Objective. To develop and evaluate the physical, chemical, and sensory quality of three white breads of baguette type by partially substitution of wheat flour (HT) with squid flour (HC). Materials and methods. The sponge dough technique was used incorporating 7,5, 10,0 and 12,5 % of HC. Physical analysis: kneading time, dough weight, dough fermentation, specific volume, bread weight (g), % loss of moisture and crumb color; chemical analysis: moisture, crude protein, ash, ether extract, crude fiber, carbohydrates, indispensable amino acids and sensory evaluation: texture, odor, color, and flavor. Results. Physical properties did not show differences (p>0.05), differences (p<0.05) for color variables in all breads as well as in chemical analysis and amino acid profile (p<0.05), the protein show an increase (15,63 % for bread with HT to 23,27 % for bread with 12.5 % of HC) and the same behavior for indispensable amino acids, that increase as inclusion of HC increases. The results of the sensory evaluation were similar (p>0.05) in control and 7,5 % of HC (I like it), for 10 and 12,5% it was indifferent. Conclusions. The inclusion of squid flour (7,5 %) in the preparation of baguette bread achieved an increase of proteins and essential amino acids that improved the nutritional quality of this food, which also presented a good acceptance according to an exploratory test with consumers(AU)

Comparison of physicochemical and volatile flavor properties of neon flying squid (Ommastrephes bartramii), jumbo squid (Dosidicus gigas), and Argentine shortfin squid (Illex argentinus) during chilled storage.

The difference and similarities in the physicochemical and volatile flavor properties were determined in neon flying squid (OB), jumbo squid (DG), and Argentine squid (IA) mantles during 8 days of chilled storage. Physicochemical analysis indicated the chilled conditions induced rapid increases in pH value, total volatile basic nitrogen (TVBN), and carbonyl and malondialdehyde (MDA) content of the three squid species. In addition, myofibrillar protein (MP) content decreased and springiness in the OB, DG, and IA mantle samples declined with the extension of storage time. Importantly, OB mantles presented less chemical stability than the other two squid samples during 8 days of chilled storage. In addition, histological observations suggest DG mantle tissues presented more compact structures than those of the other two samples. Volatile flavor analysis showed propionaldehyde, 3-pentanone, trimethylamine, 3-furanmethanol, 2-methyl butyric acid, and 2-butanone were highly abundant in the squid mantles after storage, likely resulting from decomposition, oxidation, and degradation of proteins and lipids in the squid mantle, which varied with different squid species. The findings provide insight into the performance of three squid species during chilled storage.

Giant Squid (Dosidicus gigas) Meal in Chicken Diets to Enrich Meat with n-3 Fatty Acids.

The main marketed parts of squid are the mantle, the head with tentacles, and fins. However, when the whole squid does not meet quality standards for human consumption it can be used for broiler feed. The objective of the study was to include giant squid (Dosidicus gigas) meal (GSM) in broiler rations to increase the content of the n-3 fatty acids eicosapentaenoic (EPA), docosapentaenoic (DPA), and docosahexaenoic (DHA) in chicken meat. Two hundred Ross 380 chickens, half male, half female, and one day old, were randomly distributed in a 4x2x2 factorial arrangement. The factors were the treatment (0%, 1.67%, 3.34%, and 5.01% of GSM in the diet), sex, and content of n-3 in the legs with thighs and the breasts. Each treatment had five repetitions with 10 birds each. There were no differences (p > 0.05) in the production parameters for both sexes. The contents of EPA, DPA, and DHA increased in the females and in the legs with thighs (p < 0.05) with GSM. Acceptance for the flavor and texture of meat was higher in the treatment with 1.67% GSM than in the other treatments. It is concluded that GSM is an alternative for increasing the amount of n-3 in chicken meat.

Social relationship dynamics mediate climate impacts on income inequality: evidence from the Mexican Humboldt squid fishery.

Small-scale fisheries are critically important for livelihoods around the world, particularly in tropical regions. However, climate variability and anthropogenic climate change may seriously impact small-scale fisheries by altering the abundance and distribution of target species. Social relationships between fishery users, such as fish traders, can determine how each individual responds and is affected by changes in fisheries. These informal cooperative and competitive relationships provide access, support, and incentives for fishing and affect the distribution of benefits. Yet, individuals' actions and impacts on individuals are often the primary focus of the economic analyses informing small-scale fisheries' formal management. This focus dismisses relevant social relationships. We argue that this leads to a disconnect between reality and its model representation used in formal management, which may reduce formal fisheries management's efficiency and efficacy and potentially trigger adverse consequences. Here, we examine this argument by comparing the predictions of a simple bioeconomic fishery model with those of a social-ecological model that incorporates the dynamics of cooperative relationships between fish traders. We illustrate model outcomes using an empirical case study in the Mexican Humboldt squid fishery. We find that (1) the social-ecological model with relationship dynamics substantially improves accuracy in predicting observed fishery variables to the simple bioeconomic model. (2) Income inequality outcomes are associated with changes in cooperative trade relationships. When environmental temperature is included in the model as a driver of species production dynamics, we find that climate-driven temperature variability drives a decline in catch that, in turn, reduce fishers' income. We observe an offset of this loss in income by including cooperative relationships between fish traders (oligopoly) in the model. These relationships break down following species distribution changes and result in an increase in prices fishers receive. Finally, (3) our social-ecological model simulations show that the current fishery development program, which seeks to increase fishers' income through an increase in domestic market demand, is supported by predictions from the simple bioeconomic model, may increase income inequality between fishers and traders. Our findings highlight the real and urgent need to re-think fisheries management models in the context of small-scale fisheries and climate change worldwide to encompass social relationship dynamics. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s10113-021-01747-5).

Factors affecting mercury concentrations in two oceanic cephalopods of commercial interest from the southern Caribbean.

Mercury (Hg) concentrations have significantly increased in oceans during the last century. This element accumulates in marine fauna and can reach toxic levels. Seafood consumption is the main pathway of methylmercury (MeHg) toxicity in humans. Here, we analyzed total Hg (T-Hg) concentrations in two oceanic squid species (Ommastrephes bartramii and Thysanoteuthis rhombus) of an increasing commercial interest off Martinique, French West Indies. Stable isotope ratios reveal a negative linear relationship between δ15N or δ13C in diamondback squid samples. No significant trend was observed between δ34S values and T-Hg concentrations, contrasting with the sulfate availability and sulfide abundance hypotheses. This adds to a growing body of evidence suggesting Hg methylation via sulfate-reducing bacteria is not the main mechanism driving Hg bioavailability in mesopelagic organisms. All squid samples present T-Hg levels below the maximum safe consumption limit (0.5 ppm), deeming the establishment of a commercial squid fishery in the region safe for human consumption.

Utilization of Marine Waste to Obtain ß-Chitin Nanofibers and Films from Giant Humboldt Squid Dosidicus gigas.

ß-chitin was isolated from marine waste, giant Humboldt squid Dosidicus gigas, and further converted to nanofibers by use of a collider machine under acidic conditions (pH 3). The FTIR, TGA, and NMR analysis confirmed the efficient extraction of ß-chitin. The SEM, TEM, and XRD characterization results verified that ß-chitin crystalline structure were maintained after mechanical treatment. The mean particle size of ß-chitin nanofibers was in the range between 10 and 15 nm, according to the TEM analysis. In addition, the ß-chitin nanofibers were converted into films by the simple solvent-casting and drying process at 60 °C. The obtained films had high lightness, which was evidenced by the CIELAB color test. Moreover, the films showed the medium swelling degree (250-290%) in aqueous solutions of different pH and good mechanical resistance in the range between 4 and 17 MPa, depending on film thickness. The results obtained in this work show that marine waste can be efficiently converted to biomaterial by use of mild extractive conditions and simple mechanical treatment, offering great potential for the future development of sustainable multifunctional materials for various industrial applications such as food packaging, agriculture, and/or wound dressing.

Interaction of Squid (Dosidicus giga) Mantle Protein with a Mixtures of Potato and Corn Starch in an Extruded Snack, as Characterized by FTIR and DSC.

The majority of snacks expanded by extrusion (SEE) are made with vegetable sources, to improve their nutritional content; it has been proposed to incorporate squid (Dosidicus gigas), due to its high protein content, low price and high availability. However, the interaction of proteins of animal origin with starch during extrusion causes negative effects on the sensory properties of SEE, so it is necessary to know the type of protein-carbohydrate interactions and their effect on these properties. The objective of this research was to study the interaction of proteins and carbohydrates of SEE elaborated with squid mantle, potato and corn. The nutritional composition and protein digestibility were evaluated, Fourier transform infrared (FTIR) and Differential Scanning Calorimetry (DSC) were used to study the formation of protein-starch complexes and the possible regions responsible for their interactions. The SEE had a high protein content (40-85%) and biological value (>93%). The melting temperature (Tm) was found between 145 and 225 °C; the Tm values in extruded samples are directly proportional to the squid content. The extrusion process reduced the amine groups I and II responsible for the protein-protein interaction and increased the O-glucosidic bonds, so these bonds could be responsible for the protein-carbohydrate interactions.

Thysanoteuthis rhombus Troschel, 1857 (Teuthida: Thysanoteuthidae) calamar diamante, registro en la zona costera de Ica, Perú / Thysanoteuthis rhombus Troschel, 1857 (Teuthida: Thysanoteuthidae) Diamond squid, registered from coastal area of Ica, Peru

Resumen En el presente trabajo a través de la pesca artesanal se registra por primera vez Thysanoteuthis rhombus en el litoral de Pisco y se amplía su rango de distribución en el pacífico oriental hasta Ica, Perú. Un ejemplar de 68 cm LM fue capturado entre los 13°50'00" S y 77°08'00" W a 50 metros de profundidad. Se realiza una breve descripción de la especie, con observaciones taxonómicas y comentarios sobre su distribución geográfica y pesquería.

Abstract In this paper, Thysanoteuthis rhombus is first registered in the coast of Pisco and its distribution range in the Eastern Pacific is extended to Ica, Peru. A 68 cm LM specimen was captured between 13°50'00"S and 77°08'00"W at 50 meters depth. A brief description of the species is given, with taxonomic observations and comments on its geographic distribution and fishery.

Influence of pH, ionic strength and isoascorbic acid on the gel-forming ability of Jumbo squid muscle (Dosidicus gigas).

The effect of pH and ionic strength (µ) on the extraction capacity of myofibrillar proteins from Jumbo squid mantle muscle along with the addition of isoascorbic acid (IA) in its gel-forming ability (GFA) were evaluated. The results indicate that µ had a greater impact (p < 0.05) than pH on the extraction of muscle myofibrillar proteins. The effectiveness of IA, as the precursor of dehydro-isoascorbic acid (DIA), on the oxidation of sulfhydryl groups (-SH) to disulfide bonds (-SS-) of extracted proteins at 0.6 µ was also evaluated. During the sol-gel transition the -SH groups initially present in the protein system decreased (p < 0.05) due to the combined effect of heat treatment (90 °C/30 min) and the addition of IA; however, the oxidative effect of IA reduced (p < 0.05) the GFA of Jumbo squid muscle proteins. Results also indicated that NaCl at 2.8% rather than at 2.5% during gel preparation significantly (p < 0.05) improves its GFA.

Changes in the volatile components of squid (illex argentinus) for different cooking methods via headspace-gas chromatography-ion mobility spectrometry.

Squid products are becoming more and more popular with consumers because of their high yields and nutrition, including novel textures with desirable sensory properties. However, it has not been determined whether the cooking method has effects on the flavor of the squid. In this study, the aroma and volatile substances of squid samples from different cooking methods (boiled, steamed, sous vide) were determined and analyzed by headspace-gas chromatography-ion mobility spectrometry and differentiated by using, as well, an electronic nose and sensory evaluation. A total of 43 characteristic flavor compounds were identified. Based on the signal intensity of the identified violate compounds, we established a fingerprint of heat-treated squid from different cooking methods. Due to the long-term low-temperature heating conditions under vacuum, the flavor of sous vide squid is different from steamed and boiled squid, and it has unique special flavor compounds. Different cooking methods can affect the aroma of squid, providing support for the industrial production of squid.

Morphological comparison between Doryteuthis pleii and D. sanpaulensis (Cephalopoda, Myopsida, Loliginidae) from Brazil

The distinction of squid species in the genus Doryteuthis is not easy due to their morphological similarity, lack of conspicuous specific characters, and overlap geographical occurrence. This difficulty has leading to an almost exclusive molecular approach, and a premature neglect of the morpho-anatomy. To emphasize that the squid phenotypic features can be useful to identify, as well as to perform any comparative analyses (such as taxonomy and phylogeny), two close species were selected as outset. Doryteuthis pleii and D. sanpaulensis are common sympatric squids in Brazilian waters, commonly used in fisheries, not so difficult to distinguish by external features of the adult specimens. The samples were analyzed from biometric data to dissections, and the found most expressive characters to distinguish them are the mantle-fin ratio; morphology of the tentacle club, its ratio compared to the mantle length; hectocotylus morphology and nidamental gland morphology.(AU)

Morphological comparison between Doryteuthis pleii and D. sanpaulensis (Cephalopoda, Myopsida, Loliginidae) from Brazil

The distinction of squid species in the genus Doryteuthis is not easy due to their morphological similarity, lack of conspicuous specific characters, and overlap geographical occurrence. This difficulty has leading to an almost exclusive molecular approach, and a premature neglect of the morpho-anatomy. To emphasize that the squid phenotypic features can be useful to identify, as well as to perform any comparative analyses (such as taxonomy and phylogeny), two close species were selected as outset. Doryteuthis pleii and D. sanpaulensis are common sympatric squids in Brazilian waters, commonly used in fisheries, not so difficult to distinguish by external features of the adult specimens. The samples were analyzed from biometric data to dissections, and the found most expressive characters to distinguish them are the mantle-fin ratio; morphology of the tentacle club, its ratio compared to the mantle length; hectocotylus morphology and nidamental gland morphology.

Partial Characterization of a Low-Molecular-Mass Fraction with Cryoprotectant Activity from Jumbo Squid (Dosidicus gigas) Mantle Muscle.

Freezing conditions affect fish muscle protein functionality due to its denaturation/aggregation. However, jumbo squid (Dosidicus gigas) muscle protein functionality remains stable even after freezing, probably due to the presence of low-molecular-mass compounds (LMMC) as cryoprotectants. Thus, water-soluble LMMC (<1 kDa) fraction obtained from jumbo squid muscle was evaluated by Fourier transform infrared spectrometry. From its spectra, total carbohydrates, free monosaccharides, free amino acids and ammonium chloride were determined. Cryoprotectant capacity and protein cryostability conferred by LMMC were investigated by differential scanning calorimetry. Fraction partial characterization showed that the main components are free amino acids (18.84 mg/g), carbohydrates (67.1 µg/mg) such as monosaccharides (51.1 µg/mg of glucose, fucose and arabinose in total) and ammonium chloride (220.4 µg/mg). Arginine, sarcosine and taurine were the main amino acids in the fraction. LMMC, at the mass fraction present in jumbo squid muscle, lowered the water freezing point to -1.2 °C, inhibiting recrystallization at 0.66 °C. Significant myofibrillar protein stabilization by LMMC was observed after a freeze-thaw cycle compared to control (muscle after extraction of LMMC), proving the effectiveness on jumbo squid protein muscle cryo- stability. Osmolytes in LMMC fraction inhibited protein denaturation/aggregation and ice recrystallization, maintaining the muscle structure stable under freezing conditions. LMMC conferred protein cryostability even at the very low mass fraction in the muscle.

Effect of Temperature and pH on the Secondary Structure and Denaturation Process of Jumbo Squid Hepatopancreas Cathepsin D.

BACKGROUND: Cathepsin D is a lysosomal enzyme that is found in all organisms acting in protein turnover, in humans it is present in some types of carcinomas, and it has a high activity in Parkinson's disease and a low activity in Alzheimer disease. In marine organisms, most of the research has been limited to corroborate the presence of this enzyme. It is known that cathepsin D of some marine organisms has a low thermostability and that it has the ability to have activity at very acidic pH. Cathepsin D of the Jumbo squid (Dosidicus gigas) hepatopancreas was purified and partially characterized. The secondary structure of these enzymes is highly conserved so the role of temperature and pH in the secondary structure and in protein denaturation is of great importance in the study of enzymes. The secondary structure of cathepsin D from jumbo squid hepatopancreas was determined by means of circular dichroism spectroscopy. OBJECTIVE: In this article, our purpose was to determine the secondary structure of the enzyme and how it is affected by subjecting it to different temperature and pH conditions. METHODS: Circular dichroism technique was used to measure the modifications of the secondary structure of cathepsin D when subjected to different treatments. The methodology consisted in dissecting the hepatopancreas of squid and freeze drying it. Then a crude extract was prepared by mixing 1: 1 hepatopancreas with assay buffer, the purification was in two steps; the first step consisted of using an ultrafiltration membrane with a molecular cut of 50 kDa, and the second step, a pepstatin agarose resin was used to purification the enzyme. Once the enzyme was purified, the purity was corroborated with SDS PAGE electrophoresis, isoelectric point and zymogram. Circular dichroism is carried out by placing the sample with a concentration of 0.125 mg / mL in a 3 mL quartz cell. The results were obtained in mdeg (millidegrees) and transformed to mean ellipticity per residue, using 111 g/mol molecular weight/residue as average. Secondary-structure estimation from the far-UV CD spectra was calculated using K2D Dichroweb software. RESULTS: It was found that α helix decreases at temperatures above 50 °C and above pH 4. Heating the enzyme above 70°C maintains a low percentage of α helix and increases ß sheet. Far-UV CD measurements of cathepsin D showed irreversible thermal denaturation. The process was strongly dependent on the heating rate, accompanied by a process of oligomerization of the protein that appears when the sample is heated, and maintained a certain time at this temperature. An amount typically between 3 and 4% α helix of their secondary structure remains unchanged. It is consistent with an unfolding process kinetically controlled due to the presence of an irreversible reaction. The secondary structure depends on pH, and a pH above 4 causes α helix structures to be modified. CONCLUSION: In conclusion, cathepsin D from jumbo squid hepatopancreas showed retaining up to 4% α helix at 80°C. The thermal denaturation of cathepsin D at pH 3.5 is under kinetic control and follows an irreversible model.

Effects of cold atmospheric plasma on squid proteases and gel properties of protein concentrate from squid (Argentinus ilex) mantle.

The effect of cold atmospheric plasma (CAP) on protein concentrate from squid (Argentinus ilex) mantle has been evaluated in terms of gel properties, protease inhibition, texture profile, color attributes, and water holding capacity. Different exposure time (15, 60, 120, 180, 240 and 300 s) at 60 kV have been employed. Our results indicated that protease activity decreases with increasing treatment time. The highest reduction (p < 0.05) in protease activity (64%) was observed after 240 s of CAP treatment. Texture profile analysis, color properties and water holding capacity of the treated squid gel revealed a significant increase. Protein carbonyl and sulfhydryl group contents findings' showed a significant increase in carbonyl content (about three times of the original content), while the total sulfhydryl group decreased (up to about 40%) in the crude extract. Microstructure and SDS-PAGE analysis revealed a high degree of protein aggregation in the squid gel treated with CAP.