Efficacy of Artificial Tears Based on an Extract of Artemia salina Containing Dinucleotides in a Rabbit Dry Eye Model.

(1) Background: Artemia salina is a brine shrimp containing high concentrations of dinucleotides, molecules with properties for dry eye treatment. For this reason, the purpose of the study was to evaluate the effect of the artificial tears based on an extract of Artemia salina in a rabbit dry eye model. (2) Methods: A prospective and randomized study was carried out. Twenty rabbits were divided into 4 groups (n = 5, each group): healthy rabbits, dry eye rabbits, dry eye rabbits treated with hypromellose (HPMC), and dry eye rabbits treated with Artemia salina. Dry eye was induced by the topical instillation of 0.2% benzalkonium chloride. The measurements were performed before and after the treatment for 5 consecutive days. (3) Results: The topical instillation of artificial tears containing Artemia salina showed beneficial effects on tear secretion, tear break-up time, corneal staining, the density of Goblet cells, heigh of mucin cloud secreted by these cells, and mRNA levels of IL-1ß and MMP9 in conjunctival cells. Compared with the HPMC, there was a statistically significant improvement (p < 0.05) with the Artemia salina in all the variables under study, except for the conjunctival hyperemia, density of Goblet cells, and mRNA levels of IL-6. (4) Conclusions: The potential of artificial tears based on Artemia salina as a secretagogue agent for dry eye treatment was confirmed, opening the door for future clinical trials and studies to extrapolate the findings for dry eye patients.

Embryonic cuticle from artemia cyst shell displays amyloid-like characteristics and nontoxicity after oral consumption.

Artemia cysts are the essential food product for industrial larviculture of fishes. The cyst shell protects the artemia embryo from mechanical damage, ultraviolet light, excessive water loss, thermal variation and anoxia condition. However, the underlying mechanism of such environmental protection is largely unclear. The embryonic cuticle of cyst shell mainly constitutes chitin and proteins. Absence of cyst shell proteins compromises embryo survival. In literature, there are few examples of functional amyloids where proteins adapt amyloid-like structures and act as protective covering. We hypothesized that the proteins from the embryonic cuticle of artemia cyst shell may have amyloid-like properties. Using FTIR and CD analysis, we found that proteins in embryonic cuticle have high ß-sheet secondary structures. Embryonic cuticles displayed high Congo red binding affinity and stained samples showed apple-green birefringence under polarized light, confirming the presence of amyloid-like structures. Amyloid structures have a tendency to propagate and cause amyloidosis. However, feeding of amyloid rich embryonic cuticles to zebrafish did not show any signs of discomfort or morbidity and amyloid deposition. Taken together, the study reveals that amyloid-like structures are present in embryonic cuticle of artemia cyst and their consumption does not induce amyloidosis in zebrafish.

Target enzymes are stabilized by AfrLEA6 and a gain of α-helix coincides with protection by a group 3 LEA protein during incremental drying.

Anhydrobiotic organisms accumulate late embryogenesis abundant (LEA) proteins, a family of intrinsically disordered proteins (IDPs) reported to improve cellular tolerance to water stress. Here we show that AfrLEA6, a Group 6 LEA protein only recently discovered in animals, protects lactate dehydrogenase (LDH), citrate synthase (CS) and phosphofructokinase (PFK) against damage during desiccation. In some cases, protection is enhanced by trehalose, a naturally-occurring protective solute. An open question is whether gain of secondary structure by LEA proteins during drying is a prerequisite for this stabilizing function. We used incremental drying (equilibration to a series of relative humidities, RH) to test the ability of AfrLEA2, a Group 3 LEA protein, to protect desiccation-sensitive PFK. AfrLEA2 was chosen due to its exceptional ability to protect PFK. In parallel, circular dichroism (CD) spectra were obtained for AfrLEA2 across the identical range of relative water contents. Protection of PFK by AfrLEA2, above that observed with trehalose and BSA, coincides with simultaneous gain of α-helix in AfrLEA2. At 100% RH, the CD spectrum for AfrLEA2 is typical of random coil, while at decreasing RH, the spectrum shows higher ellipticity at 191 nm and minima at 208 and 220 nm, diagnostic of α-helix. This study provides experimental evidence linking the gain of α-helix with stabilization of a target protein across a graded series of hydration states. Mechanistically, it is intriguing that certain other functions of these IDPs, like preventing aggregation of target proteins, can occur in fully hydrated cells and apparently do not require gain of α-helix.

Construction of thermally robust and porous shrimp ferritin crystalline for molecular encapsulation through intermolecular arginine-arginine attractions.

Protein colloid crystals are considered as high porous soft materials, presenting great potentials in nutrients and drug encapsulation, but protein crystal fabrication usually needs precipitant and high protein concentration. Herein, an easy implemented approach was reported for the construction of protein colloid crystals in diluted solution with shimp ferritin as building blocks by taking advantage of the strength of multiple intermolecular arginine-arginine interactions. The X-ray single-crystal structure reveals that a group of exquisite arginine-arginine interactions between two neighboring ferritin enable them self-assembly into long-range ordered protein soft materials. The arginine-arginine interactions mediate crystal generation favored at pH 9.5 with 200 mM NaCl, and the resulting colloid crystals exhibit high thermal stability (90 °C for 30 min). Importantly, the interglobular cavity in colloid crystals is three times larger in volume than that of intrinsic ferritin cavity in each unit cell, which can be used for molecular encapsulation.

Development of a novel Artemia eggshell-zirconium nanocomposite for efficient fluoride removal.

Fluoride pollution in water has attracted widespread concern worldwide. In this study, an Artemia eggshell-zirconium (Aes-Z) nanocomposite has been used for fluoride removal. Material characterization results showed that nano-ZrO2 was immobilized on the inner surface of the Artemia eggshell, and there was no pore blockage on the composite material. Various parameters influencing on the fluoride removal, including treatment time, composite dosage, pH, initial fluoride concentration, and other anions, were analyzed. The removal efficiency of the composite material was better than that of the single zirconia material. The removal percentage of fluoride reached 93% in 30 min with an initial fluoride concentration of 10 mg/L and a nanocomposite dosage of 8.0 g/L. The composite material had a high removal efficiency for fluoride in the pH region 4.0-10.0. The adsorption of fluoride was not influenced by the common anions (e.g., Cl-, SO42-, and NO3-) in water. The regeneration revealed that the Aes-Z composite material could be reused and remove fluoride effectively in four cycles. The pseudo-second-order rate model adequately represented the adsorption kinetics of the Aes-Z composite material. A possible, defluoridation mechanism of the Aes-Z composite material was also proposed. This study demonstrates that Aes-Z is a promising adsorbent material for fluoride removal.

Adsorption of sulfonamides to marine diatoms and arthropods.

Sulfonamides are frequently detected in the environment, where these compounds adsorb to soil particles and are retained in the environment. However, adsorption of sulfonamides to planktonic particles in the sea is not known. Here we demonstrate that sulfonamides adsorb to a diatom (Chaetoceros) and an arthropod (Artemia), albeit at low levels, under laboratory conditions. In both plankton, sulfamethazine (SMT) was more readily adsorbed than was sulfamethoxazole (SMX). The adsorption occurred quickly and the concentration on the plankton was stable for at least 24 h (Chaetoceros) or 5 h (Artemia). These data suggest that marine plankton may retain sulfonamides, although the adsorbed concentration per cell or individual is not high.

Efficient, fast and inexpensive bioassay to monitor benthic microalgae toxicity: Application to Ostreopsis species.

Even though HPLC-MS is commonly used to quantify the toxin content of Ostreopsis spp. cells, there is a need to develop easy-to-use toxicological tests to set thresholds during Ostreopsis spp. blooms. The crustacean Artemia has been widely used to evaluate the presence and toxicity of chemicals and biological contaminants and we anticipated that it could also be useful to test Ostreopsis spp. toxicity. Its relevance was first assessed by investigating the variability of the toxic effects among Ostreopsis spp. strains and throughout the dinoflagellate life cycle in combination with chemical analyses of the toxinic content by UHPLC-HRMS. After testing the toxicity of fractions prepared from Ostreopsis spp. cells, the known ova- and paly-toxins were not the only toxic metabolites to Artemia franciscana, indicating that other toxic compounds synthesized by Ostreopsis spp. still remain to be identified. To extend the bioassay to in situ monitoring, the toxicity of the benthic microalgal consortium was tested during a natural bloom of Ostreopsis cf. ovata in the NW Mediterranean Sea. The results highlight the accuracy and sensitivity of the ecotoxicological assay with Artemia franciscana to assess the toxicity of Ostreopsis spp. blooms.

Oral vaccination using Artemia coated with recombinant Saccharomyces cerevisiae expressing cyprinid herpesvirus-3 envelope antigen induces protective immunity in common carp (Cyprinus carpio var. Jian) larvae.

Cyprinid herpesvirus 3 (CyHV-3) is the etiological agent of koi herpersvirus disease (KHVD), which causes serious economic losses in global common carp and ornamental koi carp production of larvae as well as adult type fish. To control KHVD, vaccines against CyHV-3 utilizing different immunization routes have been developed, among them, oral vaccination is the most desirable method to prevent fish diseases occurring at the early larval stage. Here, we developed an oral subunit vaccine through the Saccharomyces cerevisiae cell surface display of CyHV-3 envelope protein pORF65, then, the recombinant yeast fed to Artemia which served as bio-encapsulation vector by subsequently feeding the common carp (Cyprinus carpio var. Jian) larvae. The fluorescent observation showed that the Artemia and S. cerevisiae could deliver intact antigen to the hindgut of carp larvae suggesting the possibility of the vector for oral immunization. On this basis, after three immunizations at a week interval, the oral vaccine induced high level of specific anti-pORF65 antibody. Meanwhile, a significant difference of immune-related genes expression occurred including cxca, IL-1ß, IFN-a1, lysozyme, IgM and CD8α between vaccined group and blank control group. In addition, 30% of relative percent survival of carp larvae after immunization was obtained post the animal infection assay, offered an certain immune protection. Our results indicated that the oral pORF65 subunit vaccine bioencapsulated in Artemia induced the activation of immune response and high level of antibodies, which could be served as an oral vaccine candidate for the prevention of CyHV-3 infection.

Live and frozen Artemia nauplii for catfish Lophiosilurus alexandri (Steindachner, 1876) larvae in different salinities.

The objective of the study was to verify the combination of live and frozen Artemia nauplii, in different salinized water, and its influence in growth and survival of the catfish Lophiosilurus alexandri (Steindachner, 1876) larvae. Larvae were exposed to a combination of two feeds and two different salinities: live Artemia nauplii + 0‰ salinity in the water; live Artemia nauplii + 2‰ salinity in the water; frozen Artemia nauplii + 0‰ salinity in the water; and frozen Artemia nauplii + 2‰ salinity in the water. The experiment was conducted in a randomized design, in a 2 × 2 factorial scheme, 5 replicates, during 15 days. The larvae final weight, weight gain, food conversion, standard and total length, biomass, survival, body width, intestinal quotient, Fulton's condition factor, and water quality were evaluated. The use of Artemia nauplii and 2 ppm salinized water for L. alexandri larviculture proved to be adequate providing greater growth. This was due to Artemia nauplii being kept alive and attractive for the cultivation of larvae in salinized water, even though salt was a stress factor. Already, the frozen Artemia nauplii was a viable food but should be used preferably in times of lack of live nauplii and without salinization of the water.

Purification and partial characterization of a new lectin from Parkia panurensis Benth. ex H.C. Hopkins seeds (Leguminosae family; Mimosoideae subfamily) and evaluation of its biological effects.

Lectins are proteins that have as one of their main characteristics recognizing and reversibly binding to carbohydrates. In this work, it was possible to purify and characterize a lectin from Parkia panurensis (Leguminosae family; Mimosoideae subfamily) seeds by a combination of the techniques: protein precipitation, along with affinity and then ion exchange chromatography using the Sepharose-mannose and diethylaminoethyl matrices, respectively. The pure lectin, called PpaL, has affinity by D-mannose, D-glucose and derivatives. PpaL was stable over a wide range of temperature and pH, and it showed an SDS-PAGE profile of only one protein band with apparent mass of 45 kDa, subsequently confirmed by mass spectrometry, and presented a molecular mass of 50,566 ± 1 Da. PAGE analysis and molecular exclusion chromatography demonstrated that PpaL is presented as a dimer in solution. Partial sequencing of the primary structure resulted in a total of 334 amino acid residues with approximately 97% similarity to Parkia biglobosa and Parkia platycephala seed lectins. PpaL was shown to be toxic against Artemia nauplii and had an LC50 of 20 µg/mL. The effects of biological activities presented by these proteins make them important biotechnological tools, demonstrating the importance of bioprospection of new lectins.

Novel pH-sensitive films based on starch/polyvinyl alcohol and food anthocyanins as a visual indicator of shrimp deterioration.

Intelligent packaging can provide better preservation and advanced convenience for consumers. In this study, corn starch and polyvinyl alcohol (PVA) were used to produce films. Two easily accessible anthocyanin sources-purple sweet potato extracts (PSPE) and red cabbage extracts (RCE), were added respectively to evaluate their potential of indicating food freshness. Film incorporated with PSPE or RCE showed distinguishable color changes in different buffers. Water vapor permeability (WVP) was not significantly (p > 0.05) affected by addition of RCE or lower level PSPE. As the extract content increased, a significant (p < 0.05) increase in thickness (from 64.0 to 97.7 or 85.5 µm with addition of PSPE or RCE, respectively), mechanical (from 7.3 to 11.3 or 9.1 MPa in TS, and from 92% to 249% or 284% in EB, added with PSPE or RCE, respectively), and thermal properties was observed, with the light transmittance reduced profoundly. PS-PSPE film exhibited bolder color, better mechanical properties and lower light transmittance than PS-RCE film at lower extract content. In addition, application for indicating shrimp freshness was conducted. The developed film presented visual color changes corresponding to TVB-N variation when shrimps were spoiled, showing its great potential as an indicator for monitoring shrimp freshness.

Estimation of bisulfate in edible plant foods, dog urine, and drugs: picomolar level detection and bio-imaging in living organisms.

In order to explore the properties of any species in solution, the actual, i.e. equilibrium concentration of the free species should be taken into account. Researchers have not paid attention to the deprotonation equilibrium between HSO4- and SO42- while probing bisulfate ion. In this study, we have addressed this concern and developed two zwitterions, CG (coumarin-integrated glycine) and CA (coumarin-integrated alanine), for the selective detection of HSO4- at a picomolar level (50 to 325 pM) with very high binding affinity (∼108 M-1) in pure water at physiological pH. The principle of HSO4- recognition was established via UV-vis and fluorescence techniques. DFT calculations suggested that the H-bonding interactions between the probes and HSO4- are the driving force for this unforeseen selectivity. The membrane penetration ability and nontoxicity of CG/CA enable them to function as staining agents in living brine shrimps and bacteria. The use of these probes for the estimation of HSO4- in various day-to-day edible foods and drugs along with urine samples is unprecedented. The significance and novelty of this study lies in the application and development of assays for estimating bisulfate in several real-world samples that are predominantly aqueous in nature, which are the first of their kind.

Bioencapsulation efficacy of sulfated galactans in adult Artemia salina for enhancing immunity in shrimp Litopenaeus vannamei.

Live food organisms like Artemia have been used for delivery of different substances such as nutrients, probiotics and immune-stimulants to aquatic animals. Previously, we reported that sulfated galactans (SG) from the red seaweed Gracilaria fisheri (G. fisheri) increased immune activity in shrimp. In the present study we further investigated the capacity and efficiency of bioencapsulation of SG in adult Artemia for delivery to tissues and potentially boosting the expression of immune genes in post larvae shrimp. SG were labelled with FITC (FITC-SG) for in vivo tracking in shrimp. Bioencapsulation of adult Artemia with FITC-SG (0-100 µg mL-1) was performed and the fluorescence intensity was detected in the gut lumen after enrichment periods of 30 min, 1 h, 2 h, 6 h and 24 h. The results showed the Artemia took up SG over time in a concentration-dependent manner. Shrimp were fed with the bioencapsulated Artemia (FITC-SG, 20 µg mL-1) and the shrimp were evaluated under a stereo-fluorescent microscope. At 24 h after administration, FITC-SG was located in gills and hepatopancreas and also bound with haemocytes. With daily SG administration, the genes IMD, IKKß were up-regulated (after 1 day) while genes dicer and proPO-I were up-regulated later (after 7 days). Moreover, continued monitoring of shrimp fed for 3 consecutive days only with SG at the dose of 0.5 mg g-1 BW showed increases in the expression of IMD, IKKß genes on day 1 and which gradually declined to normal levels on day 14, while the expression of dicer and proPO-I was increased on day 3 and remained high on day 14. These results demonstrate that bioencapsulation of SG in adult Artemia successfully delivers SG to shrimp tissues, which then bind with haemocytes and subsequently activate immune genes, and potentially increase immunity in shrimp. In addition, the present study suggests that a 3-consecutive-day regimen of SG supplemented in Artemia (0.5 mg g-1 BW) may boost and sustain the enhanced immune functions in post larvae shrimp.

Time-dependent effects of polystyrene nanoparticles in brine shrimp Artemia franciscana at physiological, biochemical and molecular levels.

Micro- (<5 mm) and nanoplastics (<1 µm) are emerging threats for marine ecosystems worldwide. Brine shrimp Artemia is recognized as a suitable model among planktonic species for studying the impact of polystyrene nanoparticles (PS NPs) through short and long-term bioassays. Our study aims to evaluate the time-dependent effects of cationic amino-modified PS-NH2 (50 nm) in A. franciscana after short- (48 h) and long-term exposure (14 days). For this purpose, nauplii were exposed to a concentration range of PS-NH2 (0.1, 1, 3 and 10 µg/mL) in natural sea water (NSW), and physiological, biochemical and molecular responses were investigated. Short-term exposure to PS-NH2 caused a decrease in nauplii growth and affected the development in a concentration-dependent manner, long-term exposure impaired the survival, but not the growth and feeding behavior. Oxidative stress was detected after short term exposure as the decrease in the activity of antioxidant enzymes, and was fully evident in the long-term as lipid peroxidation, suggesting an accumulative effect. The decrease in Cholinesterase (ChE) activity observed indicates possible neurotoxic action of PS-NH2. Also, Carboxylesterase (CbE) inhibition by PS-NH2, described for the first time in this study, anticipates potential effects in biotransformation of exogenous and endogenous compounds, being the crustacean juvenile hormone methyl farnesoate (MF) that regulates development and molting, one candidate. Furthermore, short- and long-term exposure to PS-NH2 affect the expression of genes involved in cell protection, development and molting. Overall, our results reveal that low PS-NH2 concentrations induce physiological, biochemical and molecular (changes in gene expression) alterations in Artemia, and point at their potential risk for this model organism, supporting the general concern about nanoplastics occurrences in aquatic environments and their ability to represent an ecological threat for aquatic zooplanktonic species.

An Anthracene Excimer Fluorescence Probe on Mesoporous Silica for Dual Functions of Detection and Adsorption of Mercury (II) and Copper (II) with Biological In Vivo Applications.

Dual functional activity by the same organic-inorganic hybrid material toward selective metal ion detection and its adsorption has drawn more attraction in the field of sensing. However, most of the hybrid materials in the literature are either for sensing studies or adsorption studies. In this manuscript, a fluorescent active hybrid material SiO2 @PBATPA is synthesized by covalent coupling of anthracene-based chelating ligand N,N'-(propane-1,3-diyl) bis(N-(anthracen-9-ylmethyl)-2-((3-(triethoxysilyl)propyl) amino) acetamide) (PBATPA) within the mesopores of newly synthesized cubic mesoporous silica. The synthetic strategy is designed to form an exclusively intramolecular excimer on a solid surface, which is then used as a sensory tool for selective detection of metal ions through fluorescence quenching by the destruction of excimer upon metal ion binding. The dual functions of sensing and adsorption studies show selectivity toward Hg2+ and Cu2+ among various metal ions with detection limits of 37 and 6 ppb, respectively, and adsorption capacities of 482 and 246 mg g-1 , respectively. This material can be used as a sensory cum adsorbent material in real food samples and living organisms such as the brine shrimp Artemia salina without any toxic effects from the material.

Identification and Functional Mechanism of Novel Angiotensin I Converting Enzyme Inhibitory Dipeptides from Xerocomus badius Cultured in Shrimp Processing Waste Medium.

ACE inhibitory dipeptides from Xerocomus badius fermented shrimp processing waste were isolated with ethanol, macroporous resin, chloroform, and Sephadex G-10 in sequence and identified by LC-MS/MS system coupled with electrospray ionization source. Molecular docking was performed for exploring the mechanism of their inhibitions. The results showed that the identified ACE inhibitory dipeptides were Cys-Cys and Cys-Arg with IC50 values of 4.37 ± 0.07 and 475.95 ± 0.11 µM, respectively. The difference between ACE inhibitor potency of Cys-Cys and Cys-Arg could be explained by results of molecular docking. Cys-Cys formed crucial coordination between carboxyl oxygen and Zn(II), hydrogen bonds with residues Ala354(O), Ala356(HN), and Tyr523(OH), and a bump with the residue His387(NE2) at the active site of ACE. There was no coordination, except for 5 hydrogen bonds (at residues His353, Ala354, Glu384, Glu403, and Arg522) and a bump (Glu411) between Cys-Arg and active site of ACE. These findings highlighted that Cys-Cys could be considered as a novel potent ACE inhibitor, and coordination between its carboxyl oxygen and Zn(II) played significant role in defining its ACE inhibitor potency.

Alternative mitochondrial respiratory chains from two crustaceans: Artemia franciscana nauplii and the white shrimp, Litopenaeus vannamei.

Mitochondrial ATP is synthesized by coupling between the electron transport chain and complex V. In contrast, physiological uncoupling of these processes allows mitochondria to consume oxygen at high rates without ATP synthesis. Such uncoupling mechanisms prevent reactive oxygen species overproduction. One of these mechanisms are the alternative redox enzymes from the mitochondrial respiratory chain, which may help cells to maintain homeostasis under stress independently of ATP synthesis. To date, no reports have been published on alternative redox enzymes in crustaceans mitochondria. Specific inhibitors were used to identify alternative redox enzymes in mitochondria isolated from Artemia franciscana nauplii, and the white shrimp, Litopenaeus vannamei. We report the presence of two alternative redox enzymes in the respiratory chain of A. franciscana nauplii, whose isolated mitochondria used glycerol-3-phosphate as a substrate, suggesting the existence of a glycerol-3-phosphate dehydrogenase. In addition, cyanide and octyl-gallate were necessary to fully inhibit this species' mitochondrial oxygen consumption, suggesting an alternative oxidase is present. The in-gel activity analysis confirmed that additional mitochondrial redox proteins exist in A. franciscana. A mitochondrial glycerol-3-phosphate dehydrogenase oxidase was identified by protein sequencing as part of a branched respiratory chain, and an alternative oxidase was also identified in this species by western blot. These results indicate different adaptive mechanisms from artemia to face environmental challenges related to the changing levels of oxygen concentration in seawater through their life cycles. No alternative redox enzymes were found in shrimp mitochondria, further efforts will determine the existence of an uncoupling mechanism such as uncoupling proteins.

Synthesis and in vitro Bio-activity Evaluation of N4-benzyl Substituted 5-Chloroisatin-3-thiosemicarbazones as Urease and Glycation Inhibitors.

A series of fifteen N4-benzyl substituted 5-chloroisatin-3-thiosemicarbazones 5a-o were synthesized and screened mainly for their antiurease and antiglycation effects. Lemna aequinocitalis growth and Artemia salina assays were carried out to determine their phytotoxicity and cytotoxicity potential. All the compounds proved to be extremely effective urease inhibitors, demonstrating enzyme inhibition much better than the reference inhibitor, thiourea (IC50 values 1.31 ± 0.06 to 3.24 ± 0.15 vs. 22.3 ± 1.12 µM). On the other hand, eight out of fifteen compounds tested, i.e. 5b, 5c, 5h-k, 5m and 5n were found to be potent glycation inhibitors. Of these, five viz. 5c, 5h-j and 5n proved to be exceedingly efficient, displaying glycation inhibition greater than the reference inhibitor, rutin (IC50 values 114.51 ± 1.08 to 229.94 ± 3.40 vs. 294.5 ± 1.5 µM).

Potential functions of LEA proteins from the brine shrimp Artemia franciscana - anhydrobiosis meets bioinformatics.

Late embryogenesis abundant (LEA) proteins are a large group of anhydrobiosis-associated intrinsically disordered proteins, which are commonly found in plants and some animals. The brine shrimp Artemia franciscana is the only known animal that expresses LEA proteins from three, and not only one, different groups in its anhydrobiotic life stage. The reason for the higher complexity in the A. franciscana LEA proteome (LEAome), compared with other anhydrobiotic animals, remains mostly unknown. To address this issue, we have employed a suite of bioinformatics tools to evaluate the disorder status of the Artemia LEAome and to analyze the roles of intrinsic disorder in functioning of brine shrimp LEA proteins. We show here that A. franciscana LEA proteins from different groups are more similar to each other than one originally expected, while functional differences among members of group three are possibly larger than commonly anticipated. Our data show that although these proteins are characterized by a large variety of forms and possible functions, as a general strategy, A. franciscana utilizes glassy matrix forming LEAs concurrently with proteins that more readily interact with binding partners. It is likely that the function(s) of both types, the matrix-forming and partner-binding LEA proteins, are regulated by changing water availability during desiccation.

Photoluminescent reduced graphene oxide quantum dots from latex of Calotropis gigantea for metal sensing, radical scavenging, cytotoxicity, and bioimaging in Artemia salina: A greener route.

In this work, we report the fabrication of green fluorescent reduced graphene oxide quantum dots (rGOQDs) from the latex of Calotropis gigantea by simple one-step microwave assisted greener route. The latex of Calotropis gigantea calcined at 300°C and its ethanolic extract is used for the synthesis of QDs, The rGOQDs showed particle size ranging from 2 to 8nm and it exhibited green fluorescent in longer UV region at 360-520nm. The rGOQDs graphitic nature was confirmed by RAMAN and XRD analysis. The FTIR, XPS demonstrate that presence of functional groups such as CO, COC, -OH, hence it's addressing them as rGOQDs. It is used to design the greener and economically adopted fluorescent probe for the detection of Pb2+ ions. It provides simple and appropriate for the selective and sensitive detection of Pb2+ ions in water purification process. It also trapped the free radicals and neutralized that and act as an excellent radical scavenger in DPPH radical scavenging assessment. These rGOQDs showed excellent biocompatibility on brine shrimp nauplii (Artemia salina) up to 160µg/mL for 24h incubation. Furthermore, rGOQDS were demonstrated as fluorescent bioimaging probe selectively in the inner digestion part of Artemia salina. In summary, stable, economically viable, highly biocompatible, greener method based rGOQDs were prepared for heavy metal ion detecting, radical scavenging, bioimaging applications which can play a vital role in the future nanotechnology-based biomedical field.