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Simple Summary: Freshwater ecosystems are among the most severely affected environments by species loss. In this context, biomonitoring plays a key role by providing reliable data on biological diversity and ecological status. Environmental DNA (eDNA) metabarcoding is a powerful and non-invasive alternative to traditional morphology-based sampling and identification methods. This study represents the first application of eDNA analysis to assess the invertebrate communities in three Sicilian Lakes: Poma, Piana degli Albanesi and Scanzano. A total of 27 species were identified, belonging to Phyla of Annelida, Arthropoda and Rotifera. Moreover, the analysis revealed the presence of alien species, dangerous species associated with the transmission of viral diseases, and potential new records for Sicily. These results provide a preliminary snapshot of invertebrate biodiversity at these sites, demonstrating how eDNA has the potential to complement, but not replace, traditional methods. Freshwater ecosystems are among the most severely affected environments by species loss caused by climate change and intense anthropogenic pressure. To preserve biodiversity, biomonitoring plays a key role by providing reliable data on biological diversity and ecological status. Environmental DNA (eDNA) metabarcoding has emerged as a powerful and non-invasive alternative to traditional morphology-based sampling and identification methods. This study represents the first application of eDNA analysis to assess the invertebrate communities in three Sicilian Lakes: Poma, Piana degli Albanesi and Scanzano. Water samples were collected at two points in each lake and after filtration with nitrocellulose membranes, eDNA was extracted and metabarcoding analysis was performed. A total of 27 species were identified, belonging to Phyla of Annelida, Arthropoda and Rotifera. Notably, the analysis revealed the presence of alien species (Daphnia parvula and Acanthocyclops americanus), a dangerous species associated with the transmission of viral diseases (Culex pipiens), and potential new records for Sicily (Stylaria lacustris, Platypalpus exilis, Pammene aurana, Limnephilus rhombicus). These results provide a preliminary snapshot of invertebrate biodiversity at these sites, demonstrating how eDNA has the potential to complement, but not replace, traditional methods, contributing to the assessment of ecosystem status. [ABSTRACT FROM AUTHOR]

Simple Summary: The Genomic Hybridization (CGH) of the total DNA from two individual animals labeled with two different fluorescent dyes are mapped on a target metaphase, which can be from the same species or from a different species. This approach permits identifying the pattern of distribution of repetitive sequences on chromosomes. Furthermore, it is possible to identify loss or gain of repetitive sequences among species through this approach, changes that can occur during the species evolution. These changes can be useful for delineating repetitive sequences dynamic or phylogenetic and conservation issues. The intraspecies and interspecies Comparative Genomic Hybridization (CGH) between the closely related Cebidae species, capuchin monkeys (Cebus capucinus, Sapajus apella), and the tamarins (Saguinus mystax, Leontocebus fuscicollis) was performed to analyze their genomes. In particular, this approach determines balanced and unbalanced repetitive DNA sequence distribution and reveals dynamics during evolution. Capuchin monkeys are considered the most ancestral group with conserved syntenies compared to the hypothetical ancestral New World monkeys' karyotype. Also, more derived karyotypes of phylogenetically distant species from the Saguinus and Leontocebus genera are analyzed here. The distribution of repetitive sequences has been traditionally studied through classical staining methods of cytogenetics. It has been hypothesized that repeats are species-specific and their conservation across closely related species are also common; their role in the genome has been extensively studied even though its role in speciation is not well studied and understood. The CGH shows bright signals with balanced and imbalanced DNA involving different genome regions: such as predominantly repetitive DNA at centromeric positions, and interstitial distribution with extended blocks. Cross-species CGH demonstrated the origin of some heterochromatic regions and identified apomorphic heterochromatin expansion events. The uncovered distribution of repetitive sequences is analyzed from an evolutionary perspective to elucidate the genomic dynamics of the repetitive sequences at the level of chromosomal organization. [ABSTRACT FROM AUTHOR]

In recent years, the invasive Atlantic blue crab (Callinectes sapidus) has increased its spread throughout the Mediterranean Sea, threatening native biodiversity and local economies. This study aimed to valorize C. sapidus sampled in Sicily by utilizing its exoskeleton as a source of chitosan, astaxanthin, and bio-phenolic compounds. These biomolecules were evaluated for their reducing, radical scavenging, and antitumor activity. The ferric ion reducing antioxidant power (FRAP) and the free radical scavenging activity against radical 2,2-Diphenyl-1-picrylhydrazyl (DPPH) were significantly higher for chitosan (3.16 ± 0.10 mg AAE/g and 8.1 ± 0.10 µmol TE/g). No significant differences were observed among the tested biomolecules in their activity in scavenging the radical 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). Both bio-phenolic compounds and astaxanthin exhibited dose-dependent cytotoxicity on CaCo-2 (IC50 = 12.47 and 18 µg/mL) and HepG2 (IC50 = 10.25 and 1.26 µg/mL) cell lines, while only bio-phenols showed no cytotoxic effect on differentiated CaCo-2 cells up to 20 µg/mL. These findings highlight the value of blue crab by-products in supporting a circular economy, offering a sustainable approach to managing this invasive species while providing bioactive compounds with promising medical and nutraceutical applications. [ABSTRACT FROM AUTHOR]

This study evaluates, for the first time, the reducing capacity, radical scavenger activity, and in vitro antitumor and anti-inflammatory effects of chitosan, astaxanthin, and bio-phenols extracted from the exoskeleton of Sicilian Procambarus clarkii, the most widespread species of invasive crayfish in the Mediterranean region. Among the extracted compounds, astaxanthin exhibited the highest antioxidant activity in all assays. Chitosan and polyphenols demonstrated reducing and radical scavenging activity; chitosan showed significant ferric ion reducing capacity in the FRAP test, while bio-phenolic compounds displayed notable radical scavenging activity in the DPPH and ABTS assays. Both astaxanthin and polyphenols showed dose-dependent cytotoxicity on two different cancer cell lines, with IC50 values of 1.45 µg/mL (phenolic extract) and 4.28 µg/mL (astaxanthin extract) for HepG2 cells and 2.45 µg/mL (phenolic extract) and 4.57 µg/mL (astaxanthin extract) for CaCo-2 cells. The bio-phenolic extract also showed potential anti-inflammatory effects in vitro by inhibiting nitric oxide production in inflamed RAW 264.7 macrophages, reducing the treated/control NO ratio to 77% and 74% at concentrations of 1.25 and 1.5 μg/mL, respectively. These results suggest that P. clarkii exoskeletons could be a valuable source of bioactive molecules for biomedical, pharmaceutical, and nutraceutical application while contributing to the sustainable management of this invasive species. [ABSTRACT FROM AUTHOR]

Simple Summary: Holothuria tubulosa is a widespread edible Mediterranean species of sea cucumber. In an attempt to identify new compounds that might exert an anti-diabetic effect, the aqueous extract from the coelomic fluid (CFE) of H. tubulosa was prepared and tested for its extracellular glucose-lowering activity on hepatic cells in culture. Here, we have identified some cellular and molecular aspects of the beneficial effect of the CFE that stimulates glucose consumption/uptake and glycogen storage inducing the increased synthesis and translocation to the cell surface of the glucose transporters GLUT-2 and -4. These marine-derived natural material is worth further exploration aimed at developing novel alternative agents against diabetes mellitus and beneficial supplements for the formulation of functional food. The cell-free aqueous extract from the coelomic fluid of Holothuria tubulosa was prepared and examined for its glucose-lowering effect on HepG2 cells in vitro. In particular, employing a combination of cytochemical, flow cytometric, PCR, and protein blot techniques, we evaluated its role on glucose internalization and storage and on the upregulation and surface translocation of the two glucose transporters GLUT-2 and -4. The changes in expression, synthesis, and/or activation of the GLUT2-related transcription factor hepatocyte nuclear factor-1 alpha (HNF1α) and the GLUT-4-translocation regulatory factors insulin receptor substrate-1 (IRS-1) and AKT were also studied. Our results showed the improved glucose response by HepG2 cells, leading to an evident increase in glucose consumption/uptake and glycogen storage upon exposure. Moreover, the extract induced molecular reprogramming involving the upregulation of (i) IRS1 gene expression, (ii) the transcription and translation levels of HNF1α, AKT, and GLUT-4, (iii) the phosphorylation level of AKT, (iv) the synthesis of GLUT-2 protein, and (v) the translocation of GLUT-2 and -4 transporters onto the plasma membrane. Cumulatively, our results suggest that the coelomic fluid extract from H. tubulosa can be taken into consideration for the development of novel treatment agents against diabetes mellitus. [ABSTRACT FROM AUTHOR]

Objectives: Essential hypertension is a common chronic condition that can exacerbate or complicate various neurological diseases that may necessitate neuroimaging. Given growing medical imaging costs and the need to understand relationships between population blood pressure control and neuroimaging utilization, we seek to quantify the relationship between maximum blood pressure recorded in a given year and same-year utilization of neuroimaging CT or MR in a large healthcare population. Methods: A retrospective population-based cohort study was performed by extracting aggregate data from a multi-institutional dataset of patient encounters from 2016, 2018, and 2020 using an informatics platform (Cosmos) consisting of de-duplicated data from over 140 academic and non-academic health systems, comprising over 137 million unique patients. A population-based sample of all patients with recorded blood pressures of at least 50 mmHg DBP or 90 mmHg SBP were included. Cohorts were identified based on maximum annual SBP and DBP meeting or exceeding pre-defined thresholds. For each cohort, we assessed neuroimaging CT and MR utilization, defined as the percentage of patients undergoing ≥1 neuroimaging exam of interest in the same calendar year. Results: The multi-institutional population consisted of >38 million patients for the most recent calendar year analyzed, with overall utilization of 3.8–5.1% for CT and 1.5–2.0% for MR across the study period. Neuroimaging utilization increased substantially with increasing annual maximum BP. Even a modest BP increase to 140 mmHg systolic or 90 mmHg diastolic is associated with 3-4-fold increases in MR and 5-7-fold increases in CT same-year imaging compared to BP values below 120 mmHg / 80 mmHg. Conclusion: Higher annual maximum recorded blood pressure is associated with higher same-year neuroimaging CT and MR utilization rates. These observations are relevant to public health efforts on hypertension management to mitigate costs associated with growing imaging utilization. [ABSTRACT FROM AUTHOR]

The Barcelona Tissue Bank was established from the merge of two previous multi-tissue banks. Potential donors are screened by Donor Center staff and multi-tissue retrieval is performed by specialized own teams. Tissue processing and preservation is performed in clean room facilities by specialised personnel. After quality control of both donor and all tissues results, the heart valves and vascular segments are stored until medical request. The aim of this report is to present the cardiovascular tissue activity and retrospectively evaluate the outcomes of the changes performed in last 20 years. Cardiovascular tissue from 4088 donors was received, specifically 3115 hearts and 2095 vascular segments were processed and evaluated. A total of 48% of the aortic valves, 68% of the pulmonary valves and 75% of the vascular segments were suitable for transplant. The main reason for discarding tissue was macroscopic morphology followed by microbiological results, for both valves and arteries. Altogether, 4360 tissues were distributed for transplantation: 2032 (47%) vascular segments, 1545 (35%) pulmonary valves and 781 (18%) aortic valves. The most common indication for aortic valve surgery was the treatment of endocarditis, while for pulmonary valves, it was congenital malformation reconstruction. Vascular segments were mainly used for reconstruction after ischemia. During this period, a number of changes were made with the goal of enhancing tissue quality, safety and efficacy. These improvements were achieved through the use of a new antibiotic cocktail, increasing of donor age criteria and changing the microbiological control strategy. [ABSTRACT FROM AUTHOR]

The study of bioactive molecules of marine origin has created an important bridge between biological knowledge and its applications in biotechnology and biomedicine. Current studies in different research fields, such as biomedicine, aim to discover marine molecules characterized by biological activities that can be used to produce potential drugs for human use. In recent decades, increasing attention has been paid to a particular group of marine invertebrates, the Ascidians, as they are a source of bioactive products. We describe omics data and computational methods relevant to identifying the mechanisms and processes of innate immunity underlying the biosynthesis of bioactive molecules, focusing on innovative computational approaches based on Artificial Intelligence. Since there is increasing attention on finding new solutions for a sustainable supply of bioactive compounds, we propose that a possible improvement in the biodiscovery pipeline might also come from the study and utilization of marine invertebrates' innate immunity. [ABSTRACT FROM AUTHOR]

Wine by-products, generated in large quantities and wasted with serious environmental and economic consequences, are an exceptional alternative to conventional and non-sustainable food sources. The aim of the study was to understand if these by-products could be reused in feed for broiler chickens, improving certain blood parameters important in immune responses, by testing different concentrations and administration times. The effects of the grape-seed and pomace flour of red-wine waste were evaluated. Three different diets were tested containing pomace (0%, 3%, and 6%), grape seeds (0%, 3%, and 6%), and a mix of pomace and grape seeds (0 and 3%) for 7, 21, and 42 days. Esterase, alkaline phosphatase, and peroxidase activity, along with glucose, reactive oxygen species, and glutathione levels, were evaluated in blood samples. The results showed significant changes in the parameters analyzed, especially after 21 and 42 days of administration. All diets administered showed significant effects based on exposure time and by-product concentration used. The greatest effects seem to have come from grape-seed flour. All the by-products of wine production analyzed in this study showed important antioxidant activities and can be included in the food context of farm animals in different growth phases at different concentrations. [ABSTRACT FROM AUTHOR]

Simple Summary: Freshwater ecosystems are important for global diversity and are subject to anthropogenic impacts. Knowing the biodiversity of these sites is important, and a revolutionary method to survey this is currently the use of environmental DNA (eDNA) released by organisms into the environment. In this study, eDNA evaluation was used to analyze water samples obtained from Lake Poma, Piana degli Albanesi Lake and Lake Scanzano. The results showed that by using eDNA, it was possible to provide the first snapshot of vertebrate biodiversity in these three lakes. Moreover, the results also showed that eDNA could be a useful tool to evaluate the ecology of the environment. Freshwater ecosystems play a key role in global diversity and are subject to a series of anthropic impacts, often leading to biodiversity loss. The organisms inhabiting these sites continuously release DNA into the environment through cells, excrement, gametes and/or decomposing matter; thus, evaluation of this eDNA could revolutionize the monitoring of biodiversity. In this study, environmental DNA metabarcoding was used for the first time in three Sicilian lakes: Lake Poma, Piana degli Albanesi Lake and Lake Scanzano. Results obtained provide the first snapshot of vertebrate biodiversity in these three lakes, where little is known, to provide valuable information useful for creating a baseline of knowledge regarding the biodiversity in these three lakes. Another important result was the detection of marine species, most likely due to some kind of anthropogenic contamination. Environmental DNA is a useful tool to evaluate both biodiversity and the ecological status of the environment; it has the potential to complement traditional methods, and the use of both approaches may offer a more comprehensive understanding of the ecosystem. [ABSTRACT FROM AUTHOR]

The effects of lipopolysaccharide (LPS) on Mif (macrophage migration inhibitory factor) gene expression in the pharynx (haemapoetic tissue) of Ciona robusta were investigated using quantitative reverse‐transcription PCR (qRT‐PCR) and in situ hybridisation (ISH). To verify the induction of an inflammatory response in the pharynx, a qRT‐PCR analysis was performed to evaluate the change in the expression of proinflammatory marker genes such as Mbl, Ptx‐like, Tnf‐α and Nf‐kb, which were shown to be upregulated 1 h post LPS challenge. The change in the expression of the two Mif paralogs in the pharynx was assessed before and after stimulation, and qRT‐PCR and ISH results showed that, although Mif2 and Mif2 were expressed in clusters of haemocytes in pharynx vessels, only Mif1 expression increased after LPS stimulation. This indicates that the Mif genes are differently regulated and respond to different ambient inputs that need further analysis. [ABSTRACT FROM AUTHOR]

Simple Summary: Posidonia oceanica is a widely distributed and abundant endemic seagrass in the Mediterranean area. In the attempt to identify new compounds that might inhibit specific targets in the signal transduction of liver carcinogenesis, aqueous extracts from green and brown (beached) leaves and rhizomes of the plant were prepared and tested for their cytotoxic activity on HepG2 hepatic cancer cells in culture. Here, we have identified the death-promoting mechanisms of green-leaf and rhizome extracts that involve the modulation of autophagy, apoptosis, and cell redox status, although partially differentiating conceivably due to their different compositions. These marine-derived natural materials are worth further exploration aimed at developing novel alternative prevention and/or treatment agents against liver tumors and beneficial supplements for the formulation of functional food and food-packaging material endowed with antioxidant and anticancer properties. Aqueous extracts from Posidonia oceanica's green and brown (beached) leaves and rhizomes were prepared, submitted to phenolic compound and proteomic analysis, and examined for their potential cytotoxic effect on HepG2 liver cancer cells in culture. The chosen endpoints related to survival and death were cell viability and locomotory behavior, cell-cycle analysis, apoptosis and autophagy, mitochondrial membrane polarization, and cell redox state. Here, we show that 24 h exposure to both green-leaf- and rhizome-derived extracts decreased tumor cell number in a dose–response manner, with a mean half maximal inhibitory concentration (IC50) estimated at 83 and 11.5 μg of dry extract/mL, respectively. Exposure to the IC50 of the extracts appeared to inhibit cell motility and long-term cell replicating capacity, with a more pronounced effect exerted by the rhizome-derived preparation. The underlying death-promoting mechanisms identified involved the down-regulation of autophagy, the onset of apoptosis, the decrease in the generation of reactive oxygen species, and the dissipation of mitochondrial transmembrane potential, although, at the molecular level, the two extracts appeared to elicit partially differentiating effects, conceivably due to their diverse composition. In conclusion, P. oceanica extracts merit further investigation to develop novel promising prevention and/or treatment agents, as well as beneficial supplements for the formulation of functional foods and food-packaging material with antioxidant and anticancer properties. [ABSTRACT FROM AUTHOR]

The Ascidian C. robusta is a powerful model for studying innate immunity. LPS induction activates inflammatory-like reactions in the pharynx and the expression of several innate immune genes in granulocyte hemocytes such as cytokines, for instance, macrophage migration inhibitory factors (CrMifs). This leads to intracellular signaling involving the Nf-kB signaling cascade that triggers downstream pro-inflammatory gene expression. In mammals, the COP9 (Constitutive photomorphogenesis 9) signalosome (CSN) complex also results in the activation of the NF-kB pathway. It is a highly conserved complex in vertebrates, mainly engaged in proteasome degradation which is essential for maintaining processes such as cell cycle, DNA repair, and differentiation. In the present study, we used bioinformatics and in-silico analyses combined with an in-vivo LPS exposure strategy, next-generation sequencing (NGS), and qRT-PCR to elucidate molecules and the temporal dynamics of Mif cytokines, Csn signaling components, and the Nf-κB signaling pathway in C. robusta. A qRT-PCR analysis of immune genes selected from transcriptome data revealed a biphasic activation of the inflammatory response. A phylogenetic and STRING analysis indicated an evolutionarily conserved functional link between the Mif-Csn-Nf-kB axis in ascidian C. robusta during LPS-mediated inflammation response, finely regulated by non-coding molecules such as microRNAs (miRNAs). [ABSTRACT FROM AUTHOR]

Simple Summary: Drugs released into the aquatic environment create serious problems for the organisms that live there. For this reason, the present study investigates the in vitro effects of the antibiotic sulfamethoxazole, widely found in wastewater, on the fertilization and development of the Arbacia lixula sea urchin. The results showed a significant reduction in the percentage of fertilized oocytes at the highest drug concentrations, together with an increase in anomalies and delays in the development of the embryo. Therefore, the data obtained suggest urgent intervention on the release of these drugs in order to prevent important alterations in the species' development and to preserve biodiversity. To date, drugs released into the aquatic environment are a real problem, and among antibiotics, sulfamethoxazole is the one most widely found in wastewater; thus, the evaluation of its toxicity on marine organisms is very important. This study, for the first time, investigates the in vitro effects of 4 concentrations of sulfamethoxazole (0.05 mg/L, 0.5 mg/L, 5 mg/L, 50 mg/L) on the fertilization and development of the sea urchin Arbacia lixula. The gametes were exposed to drugs in three different stages: simultaneously with, prior to, and post-fertilization. The results show a significant reduction in the percentage of fertilized oocytes at the highest drug concentrations. Moreover, an increase in anomalies and delays in embryo development following the treatment with the drug was demonstrated. Therefore, the data suggest that this antibiotic can alter the development of marine organisms, making it urgent to act to reduce their release and to determine the concentration range with the greatest impact. [ABSTRACT FROM AUTHOR]

From an environmental and economic viewpoint, it is a win-win strategy to use materials obtained from renewable resources for the production of high-performance elastomer composites. Lignin, being a renewable biomass, was employed as a functional filler material to obtain an elastomer composite with a higher degree of mechanical performance. In the presence of a suitable coupling agent, an elevated temperature was preferred for the reactive mixing of lignin with polybutadiene rubber (BR). It is quite fascinating that the mechanical performance of this composite was comparable with carbon black-filled composites. The extraordinary reinforcing behavior of lignin in the BR matrix was understood by an available model of rubber reinforcement. In rubber composite preparation, the interfacial interaction between polybutadiene rubber and lignin in the presence of a coupling agent enabled the efficient dispersion of lignin into the rubber matrix, which is responsible for the excellent mechanical properties of the rubber composites. The rubber composites thus obtained may lead to the development of a sustainable and cost-effective end product with reliable performance. This novel approach could be implemented in other type of elastomeric materials, enabling a genuine pathway toward a sustainable globe. [ABSTRACT FROM AUTHOR]

The close phylogenetic relationship between Ciona robusta and vertebrates makes it a powerful model for studying innate immunity and the evolution of immune genes. To elucidate the nature and dynamics of the immune response, the molecular mechanisms by which bacterial infection is detected and translated into inflammation and how potential pattern recognition receptors (PRRs) are involved in pathogen recognition in tunicate C. robusta (formerly known as Ciona intestinalis), we applied an approach combining bacterial infections, next-generation sequencing, qRT-PCR, bioinformatics and in silico analyses (criteria of a p-value < 0.05 and FDR < 0.05). A STRING analysis indicated a functional link between components of the Tlr/MyD88-dependent signalling pathway (Tlr2, MyD88, and Irak4) and components of the Nf-κB signalling pathway (Nf-κB, IκBα, and Ikkα) (p-value < 0.05, FDR < 0.05). A qRT-PCR analysis of immune genes selected from transcriptome data revealed Mif as more frequently expressed in the inflammatory response than inflammation mediator or effector molecules (e.g., Il-17s, Tnf-α, Tgf-β, Mmp9, Tlrs, MyD88, Irak4, Nf-κB, and galectins), suggesting close interplay between Mif cytokines and Nf-κB signalling pathway components in the biphasic activation of the inflammatory response. An in silico analyses of the 3′-UTR of Tlr2, MyD88, IκBα, Ikk, and Nf-κB transcripts showed the presence of GAIT elements, which are known to play key roles in the regulation of immune gene-specific translation in humans. These findings provide a new level of understanding of the mechanisms involved in the regulation of the C. robusta inflammatory response induced by LPS and suggest that in C. robusta, as in humans, a complex transcriptional and post-transcriptional control mechanism is involved in the regulation of several inflammatory genes. [ABSTRACT FROM AUTHOR]

In recent years, social networks, microblogs, and short message service have deeply penetrated peoples lives, and thus, chat-style text is a common phenomenon. This chat-style text has many unknown features for linguists, which can be discovered by analyzing a chat-style corpus. The process of constructing a corpus conforms to specific corpus criteria, such as representativeness, sampling, variety, and chronology. Up to now, literature does not provide specific corpus criteria for creating a chat-style-text corpus. In contrast to related work, corpus criteria for creating a chat-style corpus are provided. An exhaustive and reliable Malay chat-style text corpus is still lacking. Thus, the provided criteria are used to demonstrate the process of constructing a Twitter corpus known as the Malay Chat-style Corpus (MCC). The MCC, which has 1 million twitter messages, consists of 14,484,384 word instances, 646,807 terms and metadata, such as posting time, used twitter client application, and type of Twitter message (simple Tweet, Retweet, Reply). Furthermore, the results of the analysis of the MCC reveal characteristics of the corpus including the most frequent terms and collocations, Zipf law diagram, Twitter peak hours, and percentages of message types. Finally, representativeness of the corpus is evaluated by employing cartography and automatic language identification methods. This corpus and the process of corpus creating are valuable for researchers working in linguistics, natural language processing, and data mining. [ABSTRACT FROM AUTHOR]

High-definition optical coherence tomography ( HD- OCT) permits real-time 3D imaging of the impact of selected agents on human skin allografts. The real-time 3D HD- OCT assessment of (i) the impact on morphological and cellular characteristics of the processing of human acellular dermal matrices ( HADMs) and (ii) repopulation of HADMs in vitro by human fibroblasts and remodelling of the extracellular matrix by these cells. Four different skin decellularization methods, Dispase II/Triton X-100, Dispase II/ SDS (sodium dodecyl sulphate), NaCl/Triton X-100 and NaCl/ SDS, were analysed by HD- OCT. HD- OCT features of epidermal removal, dermo-epidermal junction ( DEJ) integrity, cellularity and dermal architecture were correlated with reflectance confocal microscopy ( RCM), histopathology and immunohistochemistry. Human adult dermal fibroblasts were in vitro seeded on the NaCl/Triton X-100 processed HADMs, cultured up to 19 days and evaluated by HD- OCT in comparison with MTT proliferation test and histology. Epidermis was effectively removed by all treatments. DEJ was best preserved after NaCl/Triton X-100 treatment. Dispase II/ SDS treatment seemed to remove all cellular debris in comparison with NaCl/Triton X-100 but disturbed the DEJ severely. The dermal micro-architectural structure and vascular spaces of (sub)papillary dermis were best preserved with the NaCl/Triton X-100. The impact on the 3D structure and vascular holes was detrimental with Dispase II/ SDS. Elastic fibre fragmentation was only observed after Dispase II incubation. HD- OCT showed that NaCl/Triton X-100 processed matrices permitted in vitro repopulation by human dermal fibroblasts (confirmed by MTT test and histology) and underwent remodelling upon increasing incubation time. Care must be taken in choosing the appropriate processing steps to maintain selected properties of the extracellular matrix in HADMs. Processing HADMs with NaCl/Triton X-100 permits in vitro the proliferation and remodelling activity of human dermal fibroblasts. HD- OCT provides unique real-time and non-invasive 3D imaging of tissue-engineered skin constructs and complementary morphological and cytological information. [ABSTRACT FROM AUTHOR]

ABSTRACT Two distinct Ciona intestinalis phenoloxidases ( CinPO1, 2) had previously been cloned and sequenced. The CinPO2 is involved in innate immunity and is expressed by inflammatory hemocytes that populate the tunic and pharynx vessels as a response to LPS inoculation. In situ hybridization and immunohistochemistry assays on histological section, showed that the expression of this gene and the produced protein are shared with oogenesis, embryogenesis and larval morphogenesis. Intriguingly, upregulation of gene transcription was found in the test cell layer that envelopes the ovary follicle, ovulated egg, and gastrula, as well as it was modulated in the zygotic nucleus of outer balstomers of 32-cell embryo, neurula presumptive epidermis tissue and larval mesenchyme. The anti- CinPO2 antibodies, specific for adult inflammatory cells, recognize epitopes in the cytoplasm of ovarian oocytes, ovulated eggs, development stages and larval mesenchyme. The overall findings disclose the precocious activation of the CinPO2 immunity-related gene, and show a developmentally programmed expression of this phenoloxidase. Furthermore, these findings support the multifunctional roles of immunity-related genes and allows us to explore new perspectives on ascidian development and immunity. J. Exp. Zool. (Mol. Dev. Evol.) 324B: 141-151, 2015. © 2015 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

A subtractive hybridization strategy for the identification of differentially expressed genes was performed between LPS-challenged and naive Ciona intestinalis. This strategy allowed the characterization of two transcripts (Ci8short and Ci8long) generated by the use of two Alternative Polyadenylation sites. The Ci8long transcript contains a protein domain with relevant homology to several components of the Receptor Transporting Protein (RTP) family not present in the Ci8short mRNA. By means of Real Time PCR and Northern Blot, the Ci8short and Ci8long transcripts showed a different pattern of gene expression with the Ci8short mRNA being strongly activated after LPS injection in the pharynx. In situ hybridization analysis demonstrated that the activation of the APA site also influenced the tissue localization of the Ci8short transcript. This analysis showed that the Ci8long mRNA was expressed in hemocytes meanwhile the Ci8short mRNA was highly transcribed also in vessel endothelial cells and in the epithelium of pharynx. These findings demonstrated that regulation of gene expression based on different polyadenylation sites is an ancestral powerful strategy influencing both the level of expression and tissue distribution of alternative transcripts. [ABSTRACT FROM AUTHOR]

Interest is increasing in biological scaffolds for tissue regeneration such as extracellular matrix membranes, developed through soft tissue decellularization. Extracellular matrix membranes were developed to heal different tendon and soft tissue lesions that are very frequent in the general population with high health-care costs and patient morbidity. The aim of this research was to evaluate a human dermal matrix (HDM) decellularized by a chemico-physical method. A primary culture of rat tenocytes was performed: tenocytes were seeded on HDM samples and on polystyrene wells as controls (CTR). Cell viability and synthetic activity were evaluated at 3 and 7 days. An in vitro microwound model was used to evaluate HDM bioactivity: after tenocyte expansion, artificial wounds were created, HDM extracts were added, and closure time and decorin synthesis were monitored histomorphometrically at 1, 4, 24, and 72 hr. A significant higher amount of collagen I was observed when cells were cultured on HDM in comparison with that on CTR (3 days: p < 0.0001; 7 days: p < 0.05). In HDM group, fibronectin synthesis was significantly higher at both experimental times ( p < 0.0001). At 3 days, proteoglycans and transforming growth factor-β1 releases were significantly higher on HDM ( p < 0.0001 and p < 0.005, respectively). The artificial microwound closure time and decorin expression were significantly enhanced by the addition of 50% HDM extract ( p < 0.05). In vitro data showed that the decellularization technique enabled the development of a matrix with adequate biological and biomechanical properties. [ABSTRACT FROM AUTHOR]

The CAP superfamily is a group of proteins that have been linked to several biological functions such as reproduction, cancer, and immune defense. A differential screening between lipopolysaccharide (LPS)-challenged and naive Ciona intestinalis has been performed to identify LPS-induced genes. This strategy has allowed the isolation of a full-length 1471-bp cDNA encoding for a 413-amino-acid protein (CiCAP). In silico analysis has shown that this polypeptide displays a modular structure with similarities to vertebrate CAP-superfamily proteins and to a collagen-binding adhesin of Streptococcus mutans. Domain organization analysis and alignment of CiCAP to other vertebrate CAP proteins have revealed a novel structure suggesting that this protein originated from a common ancestor gene that gave rise to many subfamilies of mosaic proteins with novel functions. Quantitative mRNA expression performed by real-time polymerase chain reaction analysis has demonstrated that this gene is rapidly activated in the pharynx of C. intestinalis a few hours after LPS injection. Moreover, in situ hybridization has shown that CiCAP mRNA is highly expressed by hemocytes with large granules contained inside the pharynx vessels. Thus, CiCAP represents a protein with novel structural domains involved in ascidian immune responses. [ABSTRACT FROM AUTHOR]

Linguistic knowledge plays an important role in phrase movement in statistical machine translation. To efficiently incorporate linguistic knowledge into phrase reordering, we propose a new approach: Linguistically Annotated Reordering (LAR). In LAR, we build hard hierarchical skeletons and inject soft linguistic knowledge from source parse trees to nodes of hard skeletons during translation. The experimental results on large-scale training data show that LAR is comparable to boundary word-based reordering (BWR) (Xiong, Liu, and Lin 2006), which is a very competitive lexicalized reordering approach. When combined with BWR, LAR provides complementary information for phrase reordering, which collectively improves the BLEU score significantly. To further understand the contribution of linguistic knowledge in LAR to phrase reordering, we introduce a syntax-based analysis method to automatically detect constituent movement in both reference and system translations, and summarize syntactic reordering patterns that are captured by reordering models. With the proposed analysis method, we conduct a comparative analysis that not only provides the insight into how linguistic knowledge affects phrase movement but also reveals new challenges in phrase reordering. [ABSTRACT FROM AUTHOR]

In situ hybridisation and immunohistochemistry analyses have shown that the Ciona intestinalis tumour necrosis factor alpha gene ( CiTNFα), which has been previously cloned and sequenced, is expressed either during the inflammatory pharynx response to lipopolysaccharide (LPS) or during the swimming larval phase of development. Granulocytes with large granules and compartment/morula cells are CiTNFα-producing cells in both inflamed pharynx and larvae. Pharynx vessel endothelium also takes part in the inflammatory response. Haemocyte nodules in the vessel lumen or associated with the endothelium suggest the involvement of CiTNFα in recruiting lymphocyte-like cells and promoting the differentiation of inflammatory haemocytes. Specific antibodies against a CiTNFα peptide have identified a 43-kDa cell-bound form of the protein. Observations of pharynx histological sections (at 4 and 8 h post-LPS inoculation) from naive and medium-inoculated ascidians have confirmed the CiTNFα-positive tissue response. Larval histological sections and whole-mount preparations have revealed that CiTNFα is expressed by trunk mesenchyme, preoral lobe and tunic cells, indicating CiTNFα-expressing cell immigration events and an ontogenetic role. [ABSTRACT FROM AUTHOR]

The purification, cloning, sequencing, molecular properties and expression of a fucose-binding lectin from the serum of Dicentrarchus labrax (DlFBL) have been previously reported. We now describe the distribution and expression of DlFBL during fish ontogeny. Immunohistochemistry and in situ hybridization assays were carried out at various developmental stages (from 10 days post-hatching larvae to juveniles). Another fucose-binding lectin, similar to DlFBL in biochemical, immunochemical and agglutinating properties, was extracted and purified from eggs and appeared to be localized in the embryo yolk sack residual. DlFBL was found in columnar and goblet cells of the intestinal epithelium of larvae (from 20 days post-hatching) and juveniles and in parenchymal tissue of juveniles. DlFBL mRNA and protein were detected in the intestinal epithelium and in hepatocytes. An amplification product from degenerate primers indicates that lectin isotypes with DlFBL epitopes are expressed in eggs and embryos. Whether the lectin fraction isolated from eggs and embryos includes DlFBL of maternal origin remains unclear. [ABSTRACT FROM AUTHOR]

A tumor necrosis factor-alpha (TNFα)-like gene from Ciona intestinalis (CiTNFα-like) body wall challenged with bacterial lipopolysaccharide (LPS) was cloned and sequenced 4 h after LPS inoculation. An open reading frame of 936 bp encoding a propeptide of 312 amino acids (35.4 kDa) displaying a transmembrane domain from positions 7 to 29, a TACE cleavage site, and a mature peptide domain of 185 amino acids (20.9 kDa), was determined with a predicted isoelectric point of 9.4. The phylogenetic tree based on deduced amino acid sequences of invertebrate TNF-like protein and vertebrate TNFs supported the divergence between the ascidian and vertebrate TNF families, whereas D. melanogaster Eiger A and B TNF-like sequences were distinctly separated from the chordate TNFs. Thus, the ascidian TNFα-like cytokine was upregulated by in vivo LPS challenge supporting its pro-inflammatory role. In the pharynx, increased expression levels were found following analysis by real-time polymerase chain reaction, whereas in situ hybridization assay showed positive hemocytes both in the tissue and in circulating hemocytes. Finally, Western blot with monoclonal antibodies disclosed human TNFα epitopes in a 15-kDa protein component of the hemolymph serum and in a 43-kDa protein contained in the hemocyte lysate supernatant prepared in the presence of detergents. Both soluble and hemocyte-bound CiTNFα-like protein therefore appeared to be modulated by the LPS challenge. [ABSTRACT FROM AUTHOR]

Convolution tree kernel has shown promising results in semantic role labeling (SRL). However, this kernel does not consider much linguistic knowledge in kernel design and only performs hard matching between subtrees. To overcome these constraints, this paper proposes a grammar-driven convolution tree kernel for SRL by introducing more linguistic knowledge. Compared with the standard convolution tree kernel, the proposed grammar-driven kernel has two advantages: 1) grammar-driven approximate substructure matching, and 2) grammar-driven approximate tree node matching. The two approximate matching mechanisms enable the proposed kernel to better explore linguistically motivated structured knowledge. Experiments on the CoNLL-2005 SRL shared task and the PropBank I corpus show that the proposed kernel outperforms the standard convolution tree kernel significantly. Moreover, we present a composite kernel to integrate a feature-based polynomial kernel and the proposed grammar-driven convolution tree kernel for SRL. Experimental results show that our composite kernel-based method significantly outperforms the previously best-reported ones. [ABSTRACT FROM AUTHOR]

Studies on inducible ascidian lectins may shed light on the evolutionary emergence of cytokine functions. Here, we show that the levels of opsonins, with IL1α-epitopes, increase in Ciona intestinalis hemolymph as a response to an inflammatory stimulus and, in particular, to intratunic injection of lipopolysaccharide (LPS). The inflammatory agent promptly (within 4 h) enhances Ca2+-independent serum hemagglutinating and opsonizing activities, which are both inhibited by D-galactose and D-galactosides (α-lactose, N-acetyl-D-lactosamine, thio-digalactoside), suggesting that anti-rabbit erythrocyte lectins with galectin properties are involved as opsonins. Inducible galectin molecules contain interleukin-1α (IL1α) epitopes, and their activities are specifically inhibited by anti-human recombinant IL1α antibody. Analysis by SDS-polyacrylamide gel electrophoresis has revealed that the density of the bands of several serum proteins increases within 4 h after LPS injection, correlated with the enhanced serum activity. Moreover, Western blot patterns demonstrate that several serum proteins (59, 37, 30, 23, 15 kDa) cross-react with the antibody as early as 4 h post-injection. Although we have not been able to establish whether, in adition to galectins, various types of D-galactose-specific lectins are contained in the serum, we show, for the first time in invertebrates, that galectin molecules with opsonic properties can be enhanced in response to a non-specific inflammatory stimulus, and that their release can be further stimulated by LPS. Finally, we reveal that multiple galectins share human IL1α epitopes, probably because of steric configuration and the oligomerization process. [ABSTRACT FROM AUTHOR]

Pretreatment of date palm seedlings with an hypoaggressive Fusarium isolate (AHD) protected them partially from further infection by Fusarium oxysporum f.sp. albedinis (Foa), the Bayoud disease pathogen. No mortality occurred during 2–3 months of incubation in plants pretreated with AHD, as opposed to aggressive isolate (ZAG) inoculated controls where up to 100% mortality was observed 15–30 days after inoculation. Such protection involved biochemical interactions between the host plant and AHD since no direct competition or antagonism was revealed between AHD and ZAG. The examination of the accumulation of phenolics and peroxidase activity, two parameters previously reported to be involved in date palm resistance to Foa, indicated that the response to AHD was correlated with the ability of pretreated date palm tissues to establish a faster defence response in the roots of both susceptible and resistant cultivars. Plants pretreated with AHD accumulated higher amounts of phenolics, mainly non-constitutive hydroxycinnamic acid derivatives, which play a crucial role in date palm defence against Foa, as previously described by our group. These compounds were accumulated along with the constitutive caffeoylshikimic acids. A faster induction of peroxidase activity in response to Foa was also recorded in plants pretreated with AHD. Given the multi-component nature of these induced responses, AHD could be part of integrated disease management strategies for a sustainable control of the palm tree Bayoud disease. [ABSTRACT FROM AUTHOR]

Cytochromes P450 (CYP) are enzymes responsible for the biotransformation of most endogenous and exogenous agents. The expression of each CYP is influenced by a unique combination of mechanisms and factors including genetic polymorphisms, induction by xenobiotics, and regulation by cytokines and hormones. In recent years, Ciona robusta, one of the closest living relatives of vertebrates, has become a model in various fields of biology, in particular for studying inflammatory response. Using an in vivo LPS exposure strategy, next-generation sequencing (NGS) and qRT-PCR combined with bioinformatics and in silico analyses, compared whole pharynx transcripts from naïve and LPS-exposed C. robusta, and we provide the first view of cytochrome genes expression and miRNA regulation in the inflammatory response induced by LPS in a hematopoietic organ. In C. robusta, cytochromes belonging to 2B,2C, 2J, 2U, 4B and 4F subfamilies were deregulated and miRNA network interactions suggest that different conserved and species-specific miRNAs are involved in post-transcriptional regulation of cytochrome genes and that there could be an interplay between specific miRNAs regulating both inflammation and cytochrome molecules in the inflammatory response in C. robusta. [ABSTRACT FROM AUTHOR]

The transforming growth factor-β (TGF-β) family of cytokines performs a multifunctional signaling, which is integrated and coordinated in a signaling network that involves other pathways, such as Wintless, Forkhead box-O (FOXO) and Hedgehog and regulates pivotal functions related to cell fate in all tissues. In the hematopoietic system, TGF-β signaling controls a wide spectrum of biological processes, from immune system homeostasis to the quiescence and self-renewal of hematopoietic stem cells (HSCs). Recently an important role in post-transcription regulation has been attributed to two type of ncRNAs: microRNAs and pseudogenes. Ciona robusta, due to its philogenetic position close to vertebrates, is an excellent model to investigate mechanisms of post-transcriptional regulation evolutionarily highly conserved in immune homeostasis. The combined use of NGS and bioinformatic analyses suggests that in the pharynx, the hematopoietic organ of Ciona robusta, the Tgf-β, Wnt, Hedgehog and FoxO pathways are involved in tissue homeostasis, as they are in human. Furthermore, ceRNA network interactions and 3′UTR elements analyses of Tgf-β, Wnt, Hedgehog and FoxO pathways genes suggest that different miRNAs conserved (cin-let-7d, cin-mir-92c, cin-mir-153), species-specific (cin-mir-4187, cin-mir-4011a, cin-mir-4056, cin-mir-4150, cin-mir-4189, cin-mir-4053, cin-mir-4016, cin-mir-4075), pseudogenes (ENSCING00000011392, ENSCING00000018651, ENSCING00000007698) and mRNA 3′UTR elements are involved in post-transcriptional regulation in an integrated way in C. robusta. [ABSTRACT FROM AUTHOR]

In this paper, we report on the use of amorphous lignin, a waste by-product of the paper industry, for the production of high performance carbon fibers (CF) as precursor with improved thermal stability and thermo-mechanical properties. The precursor was prepared by blending of lignin with polyacrylonitrile (PAN), which was previously dissolved in an ionic liquid. The fibers thus produced offered very high thermal stability as compared with the fiber consisting of pure PAN. The molecular compatibility, miscibility, and thermal stability of the system were studied by means of shear rheological measurements. The achieved mechanical properties were found to be related to the temperature-dependent relaxation time (consistence parameter) of the spinning dope and the diffusion kinetics of the ionic liquids from the fibers into the coagulation bath. Furthermore, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and dynamic mechanical tests (DMA) were utilized to understand in-depth the thermal and the stabilization kinetics of the developed fibers and the impact of lignin on the stabilization process of the fibers. Low molecular weight lignin increased the thermally induced physical shrinkage, suggesting disturbing effects on the semi-crystalline domains of the PAN matrix, and suppressed the chemically induced shrinkage of the fibers. The knowledge gained throughout the present paper allows summarizing a novel avenue to develop lignin-based CF designed with adjusted thermal stability. [ABSTRACT FROM AUTHOR]