Vertebrate mineralized tissues: A modular structural analysis.

Vertebrate mineralized tissues, present in bones, teeth and scales, have complex 3D hierarchical structures. As more of these tissues are characterized in 3D using mainly FIB SEM at a resolution that reveals the mineralized collagen fibrils and their organization into collagen fibril bundles, highly complex and diverse structures are being revealed. In this perspective we propose an approach to analyzing these tissues based on the presence of modular structures: material textures, pore shapes and sizes, as well as extents of mineralization. This modular approach is complimentary to the widely used hierarchical approach for describing these mineralized tissues. We present a series of case studies that show how some of the same structural modules can be found in different mineralized tissues, including in bone, dentin and scales. The organizations in 3D of the various structural modules in different tissues may differ. This approach facilitates the framing of basic questions such as: are the spatial relations between modular structures the same or similar in different mineralized tissues? Do tissues with similar sets of modules carry out similar functions or can similar functions be carried out using a different set of modular structures? Do mineralized tissues with similar sets of modules have a common developmental or evolutionary pathway? STATEMENT OF SIGNIFICANCE: 3D organization studies of diverse vertebrate mineralized tissues are revealing detailed, but often confusing details about the material textures, the arrangements of pores and differences in the extent of mineralization within a tissue. The widely used hierarchical scheme for describing such organizations does not adequately provide a basis for comparing these tissues, or addressing issues such as structural components thought to be characteristic of bone, being present in dermal tissues and so on. The classification scheme we present is based on identifying structural components within a tissue that can then be systematically compared to other vertebrate mineralized tissues. We anticipate that this classification approach will provide insights into structure-function relations, as well as the evolution of these tissues.

Mechanical, optical, chemical, and biological evaluations of fish scale-derived scaffold for corneal replacements: A systematic review.

Corneal blindness is commonly treated through corneal replacement with allogeneic corneal donors, which may face shortage. Regarding this issue, xenogeneic alternatives are explored. Fish scale-derived scaffolds (FSSs) are among the alternatives due to the lower risk of infection and abundant sources of raw materials. Unfortunately, the information about mechanical, optical, chemical, and biological performances of FSSs for corneal replacements is still scattered, as well as about the fabrication techniques. This study aims to gather scattered pieces of information about the mentioned performances and fabrication techniques of FSSs for corneal replacements. Sorted from four scientific databases and using the PRISMA checklist, eleven relevant articles are collected. FSSs are commonly fabricated using decellularization and decalcification processes, generating FSSs with parallel multilayers or crossed fibers with topographic microchannels. In the collected studies, similar mechanical properties of FSSs to native tissues are discovered, as well as good transparency, light remittance, but poorer refractive indexes than native tissues. Biological evaluations mostly discuss histology, cell proliferations, and immune responses on FSSs, while only a few studies examine the vascularization. No studies completed comprehensive evaluations on the four properties. The current progress of FSS developments demonstrates the potential of FSS use for corneal replacements.

Efficacy and side effects of bio-fabricated sardine fish scale silver nanoparticles against malarial vector Anopheles stephensi.

Mosquitoes are a great menace for humankind since they transmit pathogenic organisms causing Malaria, Dengue, Chikungunya, Elephantiasis and Japanese encephalitis. There is an urgent need to discover new and novel biological tools to mitigate mosquito-borne diseases. To develop bioinsecticides through newly developed nanotechnology is another option in the present research scenario. In this study we synthesize and characterize sardine fish scales with silver nitrate by adopting various instrumental techniques such as UV- and FTIR-spectroscopy, energy-dispersive X-ray (EDAX), X-ray diffraction analyses (XRD) and scanning electron microscopy (SEM). Toxicity bioassays were conducted with young developmental stages of mosquito vectors. Significant mortality appeared after different life stages of mosquito vectors (young larval and pupal instars were exposed to the nanomaterials). LC50 values were 13.261 ppm for young first instar larvae and 32.182 ppm for pupae. Feeding and predatory potential of G. affinis, before and after exposure to nanoparticles against mosquito larval (I & II) instars of the mosquitoes showed promising results in laboratory experiments. Feeding potential of mosquito fish without nanoparticle treatment was 79.7% and 70.55% for the first and second instar larval populations respectively. At the nanoparticle-exposed situation the predatory efficiency of mosquitofish was 94.15% and 84.3%, respectively. Antioxidant enzymes like (SOD), (CAT), and (LPO) were estimated in the gill region of sardine fish in control and experimental waters. A significant reduction of egg hatchability was evident after nanoparticle application. It became evident from this study that the nano-fabricated materials provide suitable tools to control the malaria vector Anopheles stephensi in the aquatic phase of its life cycle. This finding suggests an effective novel approach to mosquito control.

Physicochemical and Antioxidant Properties of Gelatin and Gelatin Hydrolysates Obtained from Extrusion-Pretreated Fish (Oreochromis sp.) Scales.

Fish gelatin and its hydrolysates exhibit a variety of biological characteristics, which include antihypertensive and antioxidant properties. In this study, fish gelatins were extracted from extrusion-pretreated tilapia scales, and then subjected to analyses to determine the physicochemical properties and antioxidant activity of the extracted gelatins. Our findings indicate that TSG2 (preconditioned with 1.26% citric acid) possessed the greatest extraction yield, as well as higher antioxidant activities compared with the other extracted gelatins. Hence, TSG2 was subjected to further hydrolyzation using different proteases and ultrafiltration conditions, which yielded four gelatin hydrolysates: TSGH1, TSGH2, TSGH3, and TSGH4. The results showed that TSGH4 (Pepsin + Pancreatin and ultrafiltration < 3000 Da) had a higher yield and greater antioxidant activity in comparison with the other gelatin hydrolysates. As such, TSGH4 was subjected to further fractionation using a Superdex peptide column and two-stage reverse-phase column HPLC chromatography, yielding a subfraction TSGH4-6-2-b, which possessed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity compared with the other fractions. Further LC-ESI/MS/MS analysis of TSGH4-6-2-b suggested two novel peptides (GYDEY and EPGKSGEQGAPGEAGAP), which could have potential as naturally-occurring peptides with antioxidant properties. These promising results suggest that these antioxidant peptides could have applications in food products, nutraceuticals, and cosmetics.

Lepidopteran wing scales contain abundant cross-linked film-forming histidine-rich cuticular proteins.

Scales are symbolic characteristic of Lepidoptera; however, nothing is known about the contribution of cuticular proteins (CPs) to the complex patterning of lepidopteran scales. This is because scales are resistant to solubilization, thus hindering molecular studies. Here we succeeded in dissolving developing wing scales from Bombyx mori, allowing analysis of their protein composition. We identified a distinctive class of histidine rich (His-rich) CPs (6%-45%) from developing lepidopteran scales by LC-MS/MS. Functional studies using RNAi revealed CPs with different histidine content play distinct and critical roles in constructing the microstructure of the scale surface. Moreover, we successfully synthesized films in vitro by crosslinking a 45% His-rich CP (BmorCPR152) with laccase2 using N-acetyl- dopamine or N-ß-alanyl-dopamine as the substrate. This molecular study of scales provides fundamental information about how such a fine microstructure is constructed and insights into the potential application of CPs as new biomaterials.

Protective effects of crimson snapper scales peptides against oxidative stress on Drosophila melanogaster and the action mechanism.

Peptides derived from crimson snapper scales (CSSPs) were reported to possess excellent free radical scavenging activities in vitro. In present study, the anti-aging and anti-oxidative stress effects of CSSPs were evaluated in Drosophila melanogaster models. Results showed that the addition of CSSPs in the diets of normal Drosophila could effectively extend their lifespan and improve the motor ability of aged Drosophila. Moreover, CSSPs could protect Drosophila from oxidative damage induced by H2O2, paraquat and UV irradiation. The extension of lifespan was found to be associated with the effects of CSSPs in improving the antioxidant defense system of Drosophila, manifesting as the reduction of oxidation products MDA and PCO, the elevated activities of T-SOD, CAT and GSH-Px, and the upregulated expression of antioxidant related genes after CSSPs supplemented. Furthermore, CSSPs at 6 mg/mL significantly downregulated mTOR signaling pathway and activated autophagy in aged male Drosophila, and the inhibition on mTOR activation was probably mediated by the antioxidant effects of CSSPs. Our findings suggest that CSSPs have the potential in making dietary supplements against natural aging and oxidative stress in organisms.

General and specific microscopic characteristics of the dorsal tail scales and the spines of the crest in the tuatara Sphenodon pucntatus (Reptilia; Rhynchocephalia; Sphenodontidae).

Dorsal crest scales and those of the tail spines of the tuatara (Sphenodon punctatus) represent different specializations involved in display and protection. Erection of the dorsal crest occurs in males during combat and courtship, but tail spines are not noticeably involved in these activities. In both scale derivatives corneous beta proteins (CBPs, formerly called beta-keratins) and intermediate filaments keratins (IFKs) were determined by immunolabelling. The dermis is dense with few sparse fibrocytes surrounded by collagen bundles, the latter rather randomly oriented in the crest scales. In the tail ridge scales banded collagen I fibrils form more regular, orthogonally aligned bundles of alternating layers with connections to the basal epidermal membrane. A conglomerate of dermal melanonophores and iridophores is present under the epidermis. The iridophores are the likely origin of the whitish colour of the crest. The epidermis shows a thicker beta-layer with serrated/indented corneocytes in the tail scales while the beta layer is reduced in the crest but contains CBPs. A relatively thick mesos layer is present in both scale derivatives, especially in the crest where its role, aside from limiting transpiration, is not known. The alpha-layer is formed by corneocytes with irregular perimeter and sparse desmosomal remnants. The high labelling intensity for CBPs in the beta-layer disappears in the mesos layer but occurs, albeit strongly reduced, in the alpha-layer as in the other body scales. The take-home message is that the dense dermis and its apical beta-layer strengthen mechanically the ridge spines while the crest is mainly supported by the firm but pliable and less dense or regular dermis.

Amalgamation and application of nano chitosan cross-linked with fish scales based activated carbon as an adsorbent for the removal of reactive dye (RB9).

The extensive discomfort in the expulsion of toxic pollutants even at mild concentrations has demanded the need for prompt methods for the evacuation of dyes and heavy metals. The effective method for depuration of dye from the effluent is by sorption. Chitosan is a bio-polymer which is gaining an increasing interest as one of the sorbents. It was obtained from the crab shells by undergoing several chemical processes and used as an adsorbent for dye, metal removal and also for pharmaceutical purposes. Cross linking it with other co polymers will increase the capacity of adsorption to a maximum level. Fish scales are considered to be a major waste in the food industry and since it takes a long time for decomposing it is considered to be one of the pollutants. Hence it is utilised by converting it into activated carbon by preliminary treatment and into a muffle furnace. The obtained activated carbon is combined with chitosan by using a cross linker and utilised for adsorption mechanism. To analyse the effect of chitosan cross linked with activated carbon obtained from fish scales in adsorption of dye Reactive Blue 9 (RB9) to evaluate the adsorption studies, kinetics, mass transfer studies, thermodynamics of the bio adsorbent.

Differentiating salmonid migratory ecotypes through stable isotope analysis of collagen: Archaeological and ecological applications.

The ability to distinguish between different migratory behaviours (e.g., anadromy and potamodromy) in fish can provide important insights into the ecology, evolution, and conservation of many aquatic species. We present a simple stable carbon isotope (δ13C) approach for distinguishing between sockeye (anadromous ocean migrants) and kokanee (potamodromous freshwater residents), two migratory ecotypes of Oncorhynchus nerka (Salmonidae) that is applicable throughout most of their range across coastal regions of the North Pacific Ocean. Analyses of kokanee (n = 239) and sockeye (n = 417) from 87 sites spanning the North Pacific (Russia to California) show that anadromous and potamodromous ecotypes are broadly distinguishable on the basis of the δ13C values of their scale and bone collagen. We present three case studies demonstrating how this approach can address questions in archaeology, archival, and conservation research. Relative to conventional methods for determining migratory status, which typically apply chemical analyses to otoliths or involve genetic analyses of tissues, the δ13C approach outlined here has the benefit of being non-lethal (when applied to scales), cost-effective, widely available commercially, and should be much more broadly accessible for addressing archaeological questions since the recovery of otoliths at archaeological sites is rare.

Structural color in Junonia butterflies evolves by tuning scale lamina thickness.

In diverse organisms, nanostructures that coherently scatter light create structural color, but how such structures are built remains mysterious. We investigate the evolution and genetic regulation of butterfly scale laminae, which are simple photonic nanostructures. In a lineage of buckeye butterflies artificially selected for blue wing color, we found that thickened laminae caused a color shift from brown to blue. Deletion of the optix patterning gene also altered color via lamina thickening, revealing shared regulation of pigments and lamina thickness. Finally, we show how lamina thickness variation contributes to the color diversity that distinguishes sexes and species throughout the genus Junonia. Thus, quantitatively tuning one dimension of scale architecture facilitates both the microevolution and macroevolution of a broad spectrum of hues. Because the lamina is an intrinsic component of typical butterfly scales, our findings suggest that tuning lamina thickness is an available mechanism to create structural color across the Lepidoptera.

From iridescent blues to vibrant purples, many butterflies display dazzling 'structural colors' created not by pigments but by microscopic structures that interfere with light. For instance, the scales that coat their wings can contain thin films of chitin, the substance that normally makes the external skeleton of insects. In slim layers, however, chitin can also scatter light to produce color, the way that oil can create iridescence at the surface of water. The thickness of the film, which is encoded by the genes of the butterfly, determines what color will be produced. Yet, little is known about how common thin films are in butterflies, exactly how genetic information codes for them, and how their thickness and the colors they produce can evolve. To investigate, Thayer et al. used a technique called Helium Ion Microscopy and examined the wings of ten related species of butterflies, showing that thin film structures were present across this sample. However, the different species have evolved many different structural colors over the past millions of years by changing the thickness of the films. Next, Thayer et al. showed that this evolution could be reproduced at a faster pace in the laboratory using common buckeye butterflies. These insects mostly have brown wings, but they can have specks of blue created by thin film structures. Individuals with more blue on their wings were mated and over the course of a year, the thickness of the film structures increased by 74%, leading to shiny blue butterflies. Deleting a gene called optix from the insects also led to blue wings. Optix was already known to control the patterns of pigments in butterflies, but it now appears that it controls structural colors as well. From solar panels to new fabrics, microscopic structures that can scatter light are useful in a variety of industries. Understanding how these elements exist and evolve in organisms may help to better design them for human purposes.

Varying fish scale derived hydroxyapatite bound hybrid peptide nanofiber scaffolds for potential applications in periodontal tissue regeneration.

New peptide based hybrid scaffolds were prepared by blending two different fish scale derived hydroxyapatite with functionalized peptide nanofibers for potential applications in periodontal tissue regeneration. The nanofibers were prepared by self-assembly of the newly designed peptide bolaamphiphile Bis (N-α-amido-glutamic acid) 1,7 heptane tetracarboxylate and functionalized with a segment of the tyrosine rich amylogenin peptide sequence MPLPPHPGHPGYINF followed by polygalacturnonic acid and hydroxyapatite derived from salmon or red-snapper fish scales. The binding interactions of the components of the scaffold was confirmed by FTIR spectroscopy as well as SEM imaging. Hybrids scaffolds with salmon scale derived HaP showed higher mechanical strength and Young's Modulus compared to snapper scale derived scaffolds. Our results indicated that while both the scaffolds supported cell proliferation and efficiently formed cell-scaffold matrices with gingival fibroblasts, we observed greater alignment of the cells in the case of scaffolds that contained snapper scale derived hydroxyapatite. Furthermore, higher differentiation ability into osteoblast like cells was seen in the case of the snapper scale derived HaP based scaffolds. Our studies indicate that the hybrid peptide nanofiber scaffold matrices, particularly those prepared using snapper scales may have significant utility in the development of biomaterials for periodontal tissue regeneration.

Contributions of intermolecular bonding and lubrication to the mechanical behavior of a natural armor.

Among many dermal armors, fish scales have become a source of inspiration in the pursuit of "next-generation" structural materials. Although fish scales function in a hydrated environment, the role of water and intermolecular hydrogen bonding to their unique structural behavior has not been elucidated. Water molecules reside within and adjacent to the interpeptide locations of the collagen fibrils of the elasmodine and provide lubrication to the protein molecules during deformation. We evaluated the contributions of this lubrication and the intermolecular bonding to the mechanical behavior of elasmodine scales from the Black Carp (Mylopharyngodon piceus). Scales were exposed to polar solvents, followed by axial loading to failure and the deformation mechanisms were characterized via optical mechanics. Displacement of intermolecular water molecules by liquid polar solvents caused significant (p ≤ 0.05) increases in stiffness, strength and toughness of the scales. Removal of this lubrication decreased the capacity for non-linear deformation and toughness, which results from the increased resistance to fibril rotations and sliding caused by molecular friction. The intermolecular lubrication is a key component of the "protecto-flexibility" of scales and these natural armors as a system; it can serve as an important component of biomimetic-driven designs for flexible armor systems. STATEMENT OF SIGNIFICANCE: The natural armor of fish has become a topic of substantial scientific interest. Hydration is important to these materials as water molecules reside within the interpeptide locations of the collagen fibrils of the elasmodine and provide lubrication to the protein molecules during deformation. We explored the opportunity for tuning the mechanical behavior of scales as a model for next-generation engineering materials by adjusting the extent of hydrogen bonding with polar solvents and the corresponding interpeptide molecular lubrication. Removal of this lubrication decreased the capacity for non-linear deformation and toughness due to an increase in resistance to fibril rotations and sliding as imparted by molecular friction. We show that intermolecular lubrication is a key component of the "protecto-flexibility" of natural armors and it is an essential element of biomimetic approaches to develop flexible armor systems.

Role of Surface Chemistry in the Superhydrophobicity of the Springtail Orchesella cincta (Insecta:Collembola).

Collembola are ancient arthropods living in soil with extensive exposure to dirt, bacteria, and fungi. To protect from the harsh environmental conditions and to retain a layer of air for breathing when submerged in water, they have evolved a superhydrophobic, liquid-repelling cuticle surface. The nonfouling and self-cleaning properties of springtail cuticle make it an interesting target of biomimetic materials design. Recent research has mainly focused on the intricate microstructures at the cuticle surface. Here we study the role of the cuticle chemistry for the Collembola species Orchesella cincta (Collembola, Entomobryidae). O. cincta uses a relatively simple cuticle structure with primary granules arranged to function as plastrons. In contrast to the Collembolan cuticle featuring structures on multiple length scales that is functional irrespective of surface chemistry, we found that the O. cincta cuticle loses its hydrophobic properties after being rinsed with dichloromethane. Sum frequency generation spectroscopy and time-of-flight secondary ion mass spectrometry in combination with high-resolution mass spectrometry show that a nanometer thin triacylglycerol-containing wax layer at the cuticle surface is essential for maintaining the antiwetting properties. Removal of the wax layer exposes chitin, terpenes, and lipid layers in the cuticle. With respect to biomimetic applications, the results show that, combined with a carefully chosen surface chemistry, superhydrophobicity may be achieved using a relatively unsophisticated surface structure rather than a complex, re-entrant surface structure alone.

Influence of female cuticular hydrocarbon (CHC) profile on male courtship behavior in two hybridizing field crickets Gryllus firmus and Gryllus pennsylvanicus.

BACKGROUND: The hybridizing field crickets, Gryllus firmus and Gryllus pennsylvanicus have several barriers that prevent gene flow between species. The behavioral pre-zygotic mating barrier, where males court conspecifics more intensely than heterospecifics, is important because by acting earlier in the life cycle it has the potential to prevent a larger fraction of hybridization. The mechanism behind such male mate preference is unknown. Here we investigate if the female cuticular hydrocarbon (CHC) profile could be the signal behind male courtship. RESULTS: While males of the two species display nearly identical CHC profiles, females have different, albeit overlapping profiles and some females (between 15 and 45%) of both species display a male-like profile distinct from profiles of typical females. We classified CHC females profile into three categories: G. firmus-like (F; including mainly G. firmus females), G. pennsylvanicus-like (P; including mainly G. pennsylvanicus females), and male-like (ML; including females of both species). Gryllus firmus males courted ML and F females more often and faster than they courted P females (p < 0.05). Gryllus pennsylvanicus males were slower to court than G. firmus males, but courted ML females more often (p < 0.05) than their own conspecific P females (no difference between P and F). Both males courted heterospecific ML females more often than other heterospecific females (p < 0.05, significant only for G. firmus males). CONCLUSIONS: Our results suggest that male mate preference is at least partially informed by female CHC profile and that ML females elicit high courtship behavior in both species. Since ML females exist in both species and are preferred over other heterospecific females, it is likely that this female type is responsible for most hybrid offspring production.

Development of microstructured fish scale collagen scaffolds to manufacture a tissue-engineered oral mucosa equivalent.

The present study aimed to develop a more biomimetic tissue-engineered oral mucosa equivalent comprising 1% type I tilapia scale collagen scaffold having microstructures mimicking the dermal-epidermal junction of oral mucosa and oral keratinocytes as graft materials for human use. We designed four micropattern prototypes mimicking the dermal-epidermal junction. Using a semiconductor process and soft lithography, negative molds were fabricated to develop microstructures using both polydimethylsiloxane and silicon substrates. Micropattern configurations of dermal-epidermal junctions manufactured from fish collagen consisting of a fibril network using our micropatterning system were well preserved, although the internal fibril network of the pillar pattern was sparse. Mixing 1% chondroitin sulfate with the collagen matrix minimized tissue-engineered oral mucosa equivalent contraction. Histologic examinations showed a flattening of the vertical dimensions of all microstructures and expansion of their pitches, indicating changes in the originally designed configurations. Nonetheless, histologic examinations revealed that a fully differentiated and stratified epithelial layer was developed on all scaffolds, suggesting that the microstructured fish scale collagen scaffolds have potential in the manufacturing of tissue-engineered oral mucosa equivalents for clinical use; however, enhancement of the mechanical properties of micropatterns is required. Our micropatterning technology can also apply to the development of oral mucosa in vitro models.

Physicochemical properties and hepatoprotective effects of glycated Snapper fish scale peptides conjugated with xylose via maillard reaction.

The physicochemical properties and hepatoprotective effects of fish scales peptides (FSP) and the glycated peptides conjugated with xylose via Maillard reaction (FSP-MRPs) were investigated. Results showed that the FSP was rich in oligopeptides within 2-10 amino acids, the degree of grafting of FSP-MRPs was 52.97 ± 1.58% and the antioxidant activities in vitro of FSP were improved through Maillard reaction. In order to investigate the antioxidant activities of FSP-MRPs after digestion, the simulated gastrointestinal digestion experiments of FSP and FSP-MRPs in vitro were conducted. Results indicated that the antioxidant activities of FSP and FSP-MRPs remained as stronger as before even under the digestive conditions. Furthermore, FSP-MRPs could significantly reduce the elevated activities of serum aspartate aminotransferase and alanine aminotransferase, decrease the elevated the levels of hepatic malondialdehyde and triglyceride, and inhibit the decrease of hepatic superoxide dismutase, catalase and glutathione peroxidase caused by alcohol-induced liver damage. These findings suggest that the glycated peptides formed by FSP and xylose via Maillard reaction may be potential to be exploited as a potential functional ingredient in food industry.

Synthesized bioadsorbent from fish scale for chromium (III) removal.

Presence of heavy metal in industrial wastewater is hazardous to the surrounding environment. Biosorption of heavy metal is an effective technology for the treatment of industrial wastewater. This research work has been carried out on removal of chromium (III) metal ions by employing waste fish scales as bioadsorbent. A batch adsorption process was carried out with different adsorbent dosage, solution pH and contact time. The results show the highest 99.7518 % chromium (III) metal ions at bioadsorbent dosage 0.8 g, pH of the solution 5 and contact time 90 min, initial concentration 150 mg/l chromium ion. The adsorption isotherms data fitted well with the Langmuir isotherm model with R2 = 0.9998, qmax = 18.3486 mg/g, and RL = 0.00007325. As well as pseudo-first and second kinetics model was also analyzed for the description of adsorption and found to be well fitted (R2 = 1) for adsorption kinetics. The surface properties activated fish scales and chromium loaded fish scale were investigated by scanning electron microscopy, X-ray spectroscopy, Fourier-transform infrared spectroscopy, and thermal analysis and agree with outcomes.

Bioactive peptides derived from crimson snapper and in vivo anti-aging effects on fat diet-induced high fat Drosophila melanogaster.

Crimson snapper is an important marine economy fish, and its scales are rich in collagen. Crimson snapper scale peptides (CSSPs) obtained from crimson snapper scales by enzymatic hydrolysis exhibited significantly antioxidant activities to DPPH, ABTS and hydroxyl radicals in vitro. In vivo antioxidant and anti-aging effects of CSSPs on the Drosophila melanogaster aging model constructed by lard were investigated in this study. The results showed that CSSPs could prolong the mean lifespan, 50% survival days and the maximum lifespan of Drosophila, and especially when the concentration of CSSPs was increased to 6 mg mL-1, the mean lifespan of male and female Drosophila increased by 21.2% and 26.2%, respectively, indicating the effective life extension effect of CSSPs. Meanwhile, the addition of CSSPs effectively reduced the contents of malondialdehyde (MDA) and protein carbonylation (PCO) and increased the levels of total superoxide dismutase (T-SOD) and catalase (CAT) in Drosophila, which showed a significant dose dependence. Besides, the antioxidant-related genes SOD1, SOD2 and CAT were up-regulated in Drosophila when fed with diets containing CSSPs. In conclusion, CSSPs played an antioxidant and anti-aging role by reducing the accumulation of peroxide products and improving the activity of antioxidant enzymes, as well as by up-regulating the expression of antioxidant-related genes. The results suggest that CSSPs have potential to be used in the food and healthcare industries as potential food-borne antioxidant and anti-aging agents.

Effects of gelatin-based antifreeze peptides on cell viability and oxidant stress of Streptococcus thermophilus during cold stage.

Cold stage adversely affects cell proliferation and cell viability of probiotics such as Streptococcus thermophilus in food industry, new type of cryoprotectants continues to be needed. Gelatin-based antifreeze peptide becomes a popular topic because of its cryoprotective effects on cold-stressed probiotics. In this study the effects of tilapia scales antifreeze peptides (TSAPP) on cell viability and oxidant stress of S. thermophilus during cold stage were investigated. The results showed that the percentage of viable cells was increased 10.85 folds compared with control groups. Addition of TSAPP activated the activities of ATPases, relieved the hyperpolarization of cell membrane potential and regulated the intracellular Ca2+ concentration. Furthermore, TSAPP significantly inhibited reactive oxygen species level and malonaldehyde content in cells. Under cryopreservation with TSAPP, cells of S. thermophilus maintained higher activities of antioxidant enzymes including catalase, peroxidase and total antioxidant capacity. These findings indicate that TSAPP likely offered its cellular protection by maintaining membrane integrity and alleviation of oxidative stress.

Enhanced extraction of hydroxyapatite from bighead carp (Aristichthys nobilis) scales using deep eutectic solvent.

Fish-scale waste is rich in biocompatible hydroxyapatite (HAp). In the present study, an environmentally friendly method of extracting HAp from fish-scale waste was developed in an effort to promote environmental sustainability. Deep eutectic solvents (choline chloride/glycerol, 1/2) were used to extract HAp from bighead carp (Aristichthys nobilis) scales. A relatively high extraction rate of 47.67% ± 1.81% was obtained under optimum conditions (70 °C, a solid/liquid ratio of 1/15 g/g and a 2.5 hr extraction time). The obtained HAp was characterized and its purity was determined using Fourier transform infrared spectroscopy and X-ray diffraction, respectively. The chemical composition was performed by energy-dispersive X-ray spectrometry and inductively coupled plasma-optical emission spectroscopy. Its morphology and particle size were observed using scanning electron microscopy and particle size distribution analysis. Thermogravimetric analysis was used to determine its thermal stability. Blood compatibility was determined using a hemolytic test. The results showed that this extraction yielded HAp with the irregular morphology, the higher Ca/P ratio, good thermal stability, and blood compatibility, indicating that the proposed method is an excellent alternative for the improved utilization of fish scale waste. PRACTICAL APPLICATION: Biocompatible hydroxyapatite (HAp) was extracted from fish scale (FS) waste by using an environmentally friendly deep eutectic solvent. The optimized extraction and structure characterization of extracted HAp were investigated in this study. The results showed that the extracted HAp had the irregular morphology, the higher Ca/P ratio, good thermal stability, and blood compatibility, which indicated that the proposed method was an excellent alternative to improving the utilization of FS waste.