Chemical compositions and nutritional profiles of two edible tunicate species (Halocynthia roretzi and Halocynthia aurantium).

As an abundant marine bioresource, tunicates could be exploited in the food industry. However, limited knowledge of their chemical composition and nutritional profiles prohibited further application. In this study, two common edible tunicate species, Halocynthia roretzi (HR) and Halocynthia aurantium (HA), were subjected to comprehensive composition analysis in terms of moisture, protein, lipids, cellulose, ash, amino acids, fatty acids, non-cellulose carbohydrates and minerals. Reddish HR was much bigger than purple HA with respect to body length and weight, and their moisture fell within 82.98 %-90.92 %. The non-edible outer shell part (OS) and edible internal organs part (IO) had a dry weight ratio of around 3:2 for both two species. Generally, for both HR and HA, IO was more abundant in protein and lipids. In contrast, OS had much higher cellulose contents, confirming the better suitability of IO as a nutritional seafood. IO was richer in essential amino acids and unsaturated fatty acids, while OS had more abundant saturated fatty acids. The detected non-cellulose monosugars ranged from 0.47 % to 1.18 % and indicated the presence of some sulfated glycans. IO of HR had higher contents of essential minerals, such as Cu, Zn, and Fe, while IO of HA showed a higher K content. To sum up, this study identified the chemical composition and nutritional profile variations among different tunicate species and various dissected parts, guiding the development of specific strategies to exploit tunicates for proper food applications.

Cellulose nanocrystals-based optical organohydrogel fiber with customizable iridescent colors for strain and humidity response.

Stimuli-responsive optical hydrogels are widely used in various fields including environmental sensing, optical encryption, and intelligent display manufacturing. However, these hydrogels are susceptible to water losses when exposed to air, leading to structural damage, significantly shortened service lives, and compromised durability. This study presents mechanically robust, environmentally stable, and multi-stimuli responsive optical organohydrogel fibers with customizable iridescent colors. These fibers are fabricated by incorporating tunicate cellulose nanocrystals, alginate, and acrylamide in a glycerol-water binary system. The synthesized fibers exhibit high strength (1.38 MPa), moisture retention capabilities, and elastic properties. Furthermore, a sensor based on these fibers demonstrates high- and low-temperature resistance along with stimuli-responsive characteristics, effectively detecting changes in environmental humidity and strains. Moreover, the fiber sensor demonstrates continuous, repeatable, and quantitatively predictable moisture discoloration responses across a humidity range of 11 % and 98 %. During strain sensing, the optical-organohydrogel-based sensor demonstrates a large working strain (50 %) and excellent cycling stability, underscoring its potential for effectively monitoring a wide range of intricate human motions. Overall, the synthesized fibers and their simple fabrication method can elicit new avenues for numerous related applications including the large-scale implementation of advanced wearable technology.

A novel multi-strategy ameliorated quasi-oppositional chaotic tunicate swarm algorithm for global optimization and constrained engineering applications.

Over the last few decades, a number of prominent meta-heuristic algorithms have been put forth to address complex optimization problems. However, there is a critical need to enhance these existing meta-heuristics by employing a variety of evolutionary techniques to tackle the emerging challenges in engineering applications. As a result, this study attempts to boost the efficiency of the recently introduced bio-inspired algorithm, the Tunicate Swarm Algorithm (TSA), which is motivated by the foraging and swarming behaviour of bioluminescent tunicates residing in the deep sea. Like other algorithms, the TSA has certain limitations, including getting trapped in the local optimal values and a lack of exploration ability, resulting in premature convergence when dealing with highly challenging optimization problems. To overcome these shortcomings, a novel multi-strategy ameliorated TSA, termed the Quasi-Oppositional Chaotic TSA (QOCTSA), has been proposed as an enhanced variant of TSA. This enhanced method contributes the simultaneous incorporation of the Quasi-Oppositional Based Learning (QOBL) and Chaotic Local Search (CLS) mechanisms to effectively balance exploration and exploitation. The implementation of QOBL improves convergence accuracy and exploration rate, while the inclusion of a CLS strategy with ten chaotic maps improves exploitation by enhancing local search ability around the most prospective regions. Thus, the QOCTSA significantly enhances convergence accuracy while maintaining TSA diversification. The experimentations are conducted on a set of thirty-three diverse functions: CEC2005 and CEC2019 test functions, as well as several real-world engineering problems. The statistical and graphical outcomes indicate that QOCTSA is superior to TSA and exhibits a faster rate of convergence. Furthermore, the statistical tests, specifically the Wilcoxon rank-sum test and t-test, reveal that the QOCTSA method outperforms the other competing algorithms in the domain of real-world engineering design problems.

Cellular remodeling and JAK inhibition promote zygotic gene expression in the Ciona germline.

Transcription control is a major determinant of cell fate decisions in somatic tissues. By contrast, early germline fate specification in numerous vertebrate and invertebrate species relies extensively on RNA-level regulation, exerted on asymmetrically inherited maternal supplies, with little-to-no zygotic transcription. However delayed, a maternal-to-zygotic transition is nevertheless poised to complete the deployment of pre-gametic programs in the germline. Here, we focus on early germline specification in the tunicate Ciona to study zygotic genome activation. We first demonstrate that a peculiar cellular remodeling event excludes localized postplasmic Pem-1 mRNA, which encodes the general inhibitor of transcription. Subsequently, zygotic transcription begins in Pem-1-negative primordial germ cells (PGCs), as revealed by histochemical detection of elongating RNA Polymerase II, and nascent Mef2 transcripts. In addition, we uncover a provisional antagonism between JAK and MEK/BMPRI/GSK3 signaling, which controls the onset of zygotic gene expression, following cellular remodeling of PGCs. We propose a 2-step model for the onset of zygotic transcription in the Ciona germline and discuss the significance of germ plasm dislocation and remodeling in the context of developmental fate specification.

Pulsation waves along the Ciona heart tube reverse by bimodal rhythms expressed by a remote pair of pacemakers.

The heart of ascidians (marine invertebrate chordates) has a tubular structure, and heartbeats propagate from one end to the other. The direction of pulsation waves intermittently reverses in the heart of ascidians and their relatives; however, the underlying mechanisms remain unclear. We herein performed a series of experiments to characterize the pacemaker systems in isolated hearts and their fragments, and applied a mathematical model to examine the conditions leading to heart reversals. The isolated heart of Ciona robusta autonomously generated pulsation waves at ∼20 to 25 beats min-1 with reversals at ∼1 to 10 min intervals. Experimental bisections of isolated hearts revealed that independent pacemakers resided on each side and also that their beating frequencies periodically changed as they expressed bimodal rhythms, which comprised an ∼1.25 to 5.5 min acceleration/deceleration cycle of a beating rate of between 0 and 25 beats min-1. Only fragments including 5% or shorter terminal regions of the heart tube maintained autonomous pulsation rhythms, whereas other regions did not. Our mathematical model, based on FitzHugh-Nagumo equations applied to a one-dimensional alignment of cells, demonstrated that the difference between frequencies expressed by the two independent terminal pacemakers determined the direction of propagated waves. Changes in the statuses of terminal pacemakers between the excitatory and oscillatory modes as well as in their endogenous oscillation frequencies were sufficient to lead to heart reversals. These results suggest that the directions of pulsation waves in the Ciona heart reverse according to the changing rhythms independently expressed by remotely coupled terminal pacemakers.

Marine-Derived Peptides with Anti-Hypertensive Properties: Prospects for Pharmaceuticals, Supplements, and Functional Food.

Hypertension, a major health concern linked to heart disease and premature mortality, has prompted a search for alternative treatments due to side effects of existing medications. Sustainable harvesting of low-trophic marine organisms not only enhances food security but also provides a variety of bioactive molecules, including peptides. Despite comprising only a fraction of active natural compounds, peptides are ideal for drug development due to their size, stability, and resistance to degradation. Our review evaluates the anti-hypertensive properties of peptides and proteins derived from selected marine invertebrate phyla, examining the various methodologies used and their application in pharmaceuticals, supplements, and functional food. A considerable body of research exists on the anti-hypertensive effects of certain marine invertebrates, yet many species remain unexamined. The array of assessments methods, particularly for ACE inhibition, complicates the comparison of results. The dominance of in vitro and animal in vivo studies indicates a need for more clinical research in order to transition peptides into pharmaceuticals. Our findings lay the groundwork for further exploration of these promising marine invertebrates, emphasizing the need to balance scientific discovery and marine conservation for sustainable resource use.

Genomic Resources and Annotations for a Colonial Ascidian, the Light-Bulb Sea Squirt Clavelina lepadiformis.

Ascidian embryos have been studied since the birth of experimental embryology at the end of the 19th century. They represent textbook examples of mosaic development characterized by a fast development with very few cells and invariant cleavage patterns and lineages. Ascidians belong to tunicates, the vertebrate sister group, and their study is essential to shed light on the emergence of vertebrates. Importantly, deciphering developmental gene regulatory networks has been carried out mostly in two of the three ascidian orders, Phlebobranchia and Stolidobranchia. To infer ancestral developmental programs in ascidians, it is thus essential to carry out molecular embryology in the third ascidian order, the Aplousobranchia. Here, we present genomic resources for the colonial aplousobranch Clavelina lepadiformis: a transcriptome produced from various embryonic stages, and an annotated genome. The assembly consists of 184 contigs making a total of 233.6 Mb with a N50 of 8.5 Mb and a L50 of 11. The 32,318 predicted genes capture 96.3% of BUSCO orthologs. We further show that these resources are suitable to study developmental gene expression and regulation in a comparative framework within ascidians. Additionally, they will prove valuable for evolutionary and ecological studies.

Effects of color variation and physiological state on ascidian microbiomes.

Ascidians, known for their color variation, host species-specific microbial symbiont communities. Some ascidians can also transition into a nonfiltering (resting) physiological state. Recent studies suggest that the microbial symbiont communities may vary across different physiological states and color morphs of the host. The colonial ascidian, Polyclinum constellatum, which exhibits several color morphs in the Caribbean Sea, periodically ceases its filtering activity. To investigate if color variation in P. constellatum is indicative of sibling speciation, we sequenced fragments of the ribosomal 18S rRNA and the mitochondrial cytochrome oxidase subunit I genes. Additionally, we sequenced a fragment of the 16S rRNA gene to characterize the microbial communities of two common color morphs (red and green) in colonies that were either actively filtering (active) or nonfiltering (resting). Phylogenetic analyses of both ascidian genes resulted in well-supported monophyletic clades encompassing all color variants of P. constellatum. Interestingly, no significant differences were observed among the microbial communities of the green and red morphs, suggesting that color variation in this species is a result of intraspecific variation. However, the host's physiological state significantly influenced the microbial community structure. Nonfiltering (resting) colonies hosted higher relative abundances of Kiloniella (Alphaproteobacteria) and Fangia (Gammaproteobacteria), while filtering colonies hosted more Reugeria (Alphaproteobacteria) and Endozoicomonas (Gammaproteobacteria). This study demonstrates that microbial symbiont communities serve as reliable indicators of the taxonomic state of their host and are strongly influenced by the host's feeding condition.

Preparation and characterization of a novel high barrier mulching film with tunicate cellulose nanocrystals/sodium alginate/alkali lignin.

In this study, the base film (CSL) was prepared by blending tunicate cellulose nanocrystals (TCNCs) extracted from tunicate shells, with sodium alginate (SA) and alkali lignin (AL). Then, the mulching film (CSL-WK) was prepared using water-borne polyurethane (WPU) as binder to install low-energy Kaolin on the surface of CSL film. The influences of composition with different concentrations on mechanical properties were studied. The tensile strength and elongation at break of CSL-WK film could reach 86.58 MPa and 50.49 %, respectively. The mulching films were characterized by degradability test, SEM, FTIR, and TGA. TCNCs had good compatibility with SA and AL, and a rough structure was formed on the surface of the film to improve the hydrophobicity. The barrier properties, including ultraviolet resistance, water contact angle, water vapor permeability, water retention, and flame retardancy, were tested. The results showed that CSL-WK films could block 97 % of ultraviolet light, reduce about 25 % of soil water loss, and self-extinguish within 7 s of open flame ignition. Note that the secondary spraying method significantly improved the barrier property of films. This study lays a foundation for the preparation of ecologically friendly, biodegradable, and high barrier mulching film, and expands the application of marine resources.

Geography and developmental plasticity shape post-larval thermal tolerance in the golden star tunicate, Botryllus schlosseri.

Local adaptation and phenotypic plasticity play key roles in mediating organisms' ability to respond to spatiotemporal variation in temperature. These two processes often act together to generate latitudinal or elevational clines in acute temperature tolerance. Phenotypic plasticity is also subject to local adaptation, with the expectation that populations inhabiting more variable environments should exhibit greater phenotypic plasticity of thermal tolerance. Here we examine the potential for local adaptation and developmental plasticity of thermal tolerance in the widespread invasive tunicate Botryllus schlosseri. By comparing five populations across a thermal gradient spanning 4.4° of latitude in the northwest Atlantic, we demonstrate that warmer populations south of the Gulf of Maine exhibit significantly increased (∼0.2 °C) post-larval temperature tolerance relative to the colder populations within it. We also show that B. schlosseri post-larvae possess a high degree of developmental plasticity for this trait, shifting their median temperature of survival (LT50) upwards by as much as 0.18 °C per 1 °C increase in environmental temperature. Lastly, we found that populations vary in their degrees of developmental plasticity, with populations that experience more pronounced short-term temperature variability exhibiting greater developmental plasticity, suggesting the local adaptation of developmental plasticity. By comparing the thermal tolerance of populations across space and through time, we demonstrate how geography and developmental plasticity have shaped thermal tolerance in B. schlosseri. These results help inform our understanding of how species are able to adjust their thermal physiology in new environments, including those encountered during invasion and under increasingly novel climate conditions.

Di-aldehyde tunicate cellulose nanocrystal (D-tCNC) aerogels for drug delivery: Effect of D-tCNC composition on aerogel structure and release properties.

Aerogels as drug carriers have the characteristics of a large specific surface area, high porosity and an elastic skeleton structure. Compared with traditional drug carriers, the use of aerogels as drug carriers can avoid the complexity of drug delivery and improve the efficiency of drug loading. In this work, the oxidation of tunicate cellulose nanocrystals (tCNCs) with NaIO4 was used to prepare di-aldehyde tunicate cellulose nanocrystals (D-tCNCs). Tetracycline (TC) was used as a drug model and pH-responsive drug-loaded aerogels were prepared by the Schiff base reaction between TC and the aldehyde group on D-tCNCs. The chemical structure, crystallinity, morphology, compression properties, porosity, swelling rate and drug loading properties were investigated by FT-IR, XRD, SEM and universal testing machines. The results showed that the porosity and equilibrium swelling ratio of the D-tCNC-TC aerogels were 95.87 % and 17.52 g/g, respectively. The stress of the D-tCNC-TC aerogel at 15 % compression was 0.07 MPa. Moreover, the analysis of drug-loaded aerogels indicated that the drug loading and encapsulation rates of D-tCNC-TC aerogels were 16.86 % and 78.75 %, respectively. In in vitro release experiments, the cumulative release rate of drug-loaded aerogel at pH = 1.2 and pH = 7.4 was 87.5 % and 79.3 %, respectively. These results indicated that D-tCNC-TC aerogels have good drug loading capacity and are pH-responsive in the pH range of 1.2 to 7.4. The prepared D-tCNC-TC aerogels are expected to be applied in drug delivery systems.

Chemical and microbial diversity of a tropical intertidal ascidian holobiont.

The tropical ascidian Eudistoma vannamei, endemic to the northeastern coast of Brazil, is considered a prolific source of secondary metabolites and hosts Actinomycetota that produce bioactive compounds. Herein, we used an omics approach to study the ascidian as a holobiont, including the microbial diversity through 16S rRNA gene sequencing and metabolite production using mass spectrometry-based metabolomics. Gene sequencing analysis revealed all samples of E. vannamei shared about 50% of the observed ASVs, and Pseudomonadota (50.7%), Planctomycetota (9.58%), Actinomycetota (10.34%), Bacteroidota (12.05%) were the most abundant bacterial phyla. Analysis of tandem mass spectrometry (MS/MS) data allowed annotation of compounds, including phospholipids, amino acids, and pyrimidine alkaloids, such as staurosporine, a member of a well-known chemical class recognized as a microbial metabolite. Isolated bacteria, mainly belonging to Streptomyces and Micromonospora genera, were cultivated and extracted with ethyl acetate. MS/MS analysis of bacterial extracts allowed annotation of compounds not detected in the ascidian tissue, including marineosin and dihydroergotamine, yielding about 30% overlapped ions between host and isolated bacteria. This study reveals E. vannamei as a rich source of microbial and chemical diversity and, furthermore, highlights the importance of omic tools for a comprehensive investigation of holobiont systems.

Functional specialization of Aurora kinase homologs during oogenic meiosis in the tunicate Oikopleura dioica.

A single Aurora kinase found in non-vertebrate deuterostomes is assumed to represent the ancestor of vertebrate Auroras A/B/C. However, the tunicate Oikopleura dioica, a member of the sister group to vertebrates, possesses two Aurora kinases (Aurora1 and Aurora2) that are expressed in proliferative cells and reproductive organs. Previously, we have shown that Aurora kinases relocate from organizing centers to meiotic nuclei and were enriched on centromeric regions as meiosis proceeds to metaphase I. Here, we assessed their respective functions in oogenic meiosis using dsRNA interferences. We found that Aurora1 (Aur1) was involved in meiotic spindle organization and chromosome congression, probably through the regulation of microtubule dynamics, whereas Aurora2 (Aur2) was crucial for chromosome condensation and meiotic spindle assembly. In vitro kinase assays showed that Aur1 and Aur2 had comparable levels of kinase activities. Using yeast two-hybrid library screening, we identified a few novel interaction proteins for Aur1, including c-Jun-amino-terminal kinase-interacting protein 4, cohesin loader Scc2, and mitochondrial carrier homolog 2, suggesting that Aur1 may have an altered interaction network and participate in the regulation of microtubule motors and cohesin complexes in O. dioica.

Development and Validation of a Cyber-Physical System Leveraging EFDPN for Enhanced WSN-IoT Network Security.

In the current digital era, Wireless Sensor Networks (WSNs) and the Internet of Things (IoT) are evolving, transforming human experiences by creating an interconnected environment. However, ensuring the security of WSN-IoT networks remains a significant hurdle, as existing security models are plagued with issues like prolonged training durations and complex classification processes. In this study, a robust cyber-physical system based on the Emphatic Farmland Fertility Integrated Deep Perceptron Network (EFDPN) is proposed to enhance the security of WSN-IoT. This initiative introduces the Farmland Fertility Feature Selection (F3S) technique to alleviate the computational complexity of identifying and classifying attacks. Additionally, this research leverages the Deep Perceptron Network (DPN) classification algorithm for accurate intrusion classification, achieving impressive performance metrics. In the classification phase, the Tunicate Swarm Optimization (TSO) model is employed to improve the sigmoid transformation function, thereby enhancing prediction accuracy. This study demonstrates the development of an EFDPN-based system designed to safeguard WSN-IoT networks. It showcases how the DPN classification technique, in conjunction with the TSO model, significantly improves classification performance. In this research, we employed well-known cyber-attack datasets to validate its effectiveness, revealing its superiority over traditional intrusion detection methods, particularly in achieving higher F1-score values. The incorporation of the F3S algorithm plays a pivotal role in this framework by eliminating irrelevant features, leading to enhanced prediction accuracy for the classifier, marking a substantial stride in fortifying WSN-IoT network security. This research presents a promising approach to enhancing the security and resilience of interconnected cyber-physical systems in the evolving landscape of WSN-IoT networks.

Tunicates as Sources of High-Quality Nutrients and Bioactive Compounds for Food/Feed and Pharmaceutical Applications: A Review.

Tunicates are widely distributed worldwide and are recognized as abundant marine bioresources with many potential applications. In this review, state-of-the-art studies on chemical composition analyses of various tunicate species were summarized; these studies confirmed that tunicates contain nutrients similar to fish (such as abundant cellulose, protein, and ω-3 fatty acid (FA)-rich lipids), indicating their practical and feasible uses for food or animal feed exploration. However, the presence of certain toxic elements should be evaluated in terms of safety. Moreover, recent studies on bioactive substances extracted from tunicates (such as toxins, sphingomyelins, and tunichromes) were analyzed, and their biological properties were comprehensively reviewed, including antimicrobial, anticancer, antioxidant, antidiabetic, and anti-inflammatory activities. In addition, some insights and prospects for the future exploration of tunicates are provided which are expected to guide their further application in the food, animal feed, and pharmaceutical industries. This review is critical to providing a new pathway for converting the common pollution issues of hydroponic nutrients into valuable marine bioresources.

Using CRISPR/Cas9 to identify genes required for mechanosensory neuron development and function.

Tunicates are marine, non-vertebrate chordates that comprise the sister group to the vertebrates. Most tunicates have a biphasic lifecycle that alternates between a swimming larva and a sessile adult. Recent advances have shed light on the neural basis for the tunicate larva's ability to sense a proper substrate for settlement and initiate metamorphosis. Work in the highly tractable laboratory model tunicate Ciona robusta suggests that sensory neurons embedded in the anterior papillae transduce mechanosensory stimuli to trigger larval tail retraction and initiate the process of metamorphosis. Here, we take advantage of the low-cost and simplicity of Ciona by using tissue-specific CRISPR/Cas9-mediated mutagenesis to screen for genes potentially involved in mechanosensation and metamorphosis, in the context of an undergraduate 'capstone' research course. This small screen revealed at least one gene, Vamp1/2/3, which appears crucial for the ability of the papillae to trigger metamorphosis. We also provide step-by-step protocols and tutorials associated with this course, in the hope that it might be replicated in similar CRISPR-based laboratory courses wherever Ciona are available.

De novo genome assembly and comparative genomics for the colonial ascidian Botrylloides violaceus.

Ascidians have the potential to reveal fundamental biological insights related to coloniality, regeneration, immune function, and the evolution of these traits. This study implements a hybrid assembly technique to produce a genome assembly and annotation for the botryllid ascidian, Botrylloides violaceus. A hybrid genome assembly was produced using Illumina, Inc. short and Oxford Nanopore Technologies long-read sequencing technologies. The resulting assembly is comprised of 831 contigs, has a total length of 121 Mbp, N50 of 1 Mbp, and a BUSCO score of 96.1%. Genome annotation identified 13 K protein-coding genes. Comparative genomic analysis with other tunicates reveals patterns of conservation and divergence within orthologous gene families even among closely related species. Characterization of the Wnt gene family, encoding signaling ligands involved in development and regeneration, reveals conserved patterns of subfamily presence and gene copy number among botryllids. This supports the use of genomic data from nonmodel organisms in the investigation of biological phenomena.

Multifunctional hemostatic polysaccharide-based sponge enhanced by tunicate cellulose: A promising approach for photothermal antibacterial activity and accelerated wound healing.

Fast and effective hemostasis and protection against wound infection play a crucial role in trauma care. In this study, a sponge scaffold with a self-expanding interpenetrating macropore structure was designed via two-step cross-linking method for hemostasis and photothermal antimicrobial activity. Oxidized Konjac glucomannan (OKGM) and chitosan (CS) were crosslinked once to form a dynamic covalent bonding network, and a basic three-dimensional fiber porous network framework was constructed by uniformly dispersing Tunicate nanocellulose (TCNCs). Secondary crosslinking introduced Polydopamine (PDA NPs) into the sponge, while dynamic hydrogen bonds were interleaved to stabilize the frame. PDA NPs enhanced the sponge's antibacterial and antioxidant properties due to its good photothermal conversion efficiency and oxygen radical scavenging ability. Compared to cotton gauze and gelatin sponges, the composite sponges showed superior blood cell adhesion and platelet activation. In tests on rat liver trauma models, composite sponges showed shorter hemostasis time (12 ± 2.17 s) and less blood loss (0.1 ± 0.052 g). Sponges can protect wound tissue through their adhesion properties. In the full-thickness wound model infected with S. aureus, the composite sponge accelerated wound healing. Overall, this composite sponge has great potential for clinical use as a wound dressing.