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Sensing of human immunodeficiency virus type 1 (HIV-1) DNA is mediated by the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) signaling axis. Signal transduction and regulation of this cascade is achieved by post-translational modifications. Here we show that cGAS-STING-dependent HIV-1 sensing requires interferon-stimulated gene 15 (ISG15). ISG15 deficiency inhibits STING-dependent sensing of HIV-1 and STING agonist-induced antiviral response. Upon external stimuli, STING undergoes ISGylation at residues K224, K236, K289, K347, K338, and K370. Inhibition of STING ISGylation at K289 suppresses STING-mediated type Ⅰ interferon induction by inhibiting its oligomerization. Of note, removal of STING ISGylation alleviates gain-of-function phenotype in STING-associated vasculopathy with onset in infancy (SAVI). Molecular modeling suggests that ISGylation of K289 is an important regulator of oligomerization. Taken together, our data demonstrate that ISGylation at K289 is crucial for STING activation and represents an important regulatory step in DNA sensing of viruses and autoimmune responses. [Display omitted] • STING is modified by ISG15 • ISGylation of STING at K289 facilitates its oligomerization • ISG15 deficiency impairs DNA sensing • SAVI-STING requires ISGylation for constitutive activity Lin et al. demonstrate that STING is ISGylated in response to external stimuli. K289-linked ISGylation of STING is essential for interferon induction and STING-dependent autoimmune disease. [ABSTRACT FROM AUTHOR]

Inherited and acquired genomic abnormalities are known to cause genetic diseases and contribute to cancer formation. Recent studies demonstrated a substantial mutational load in mouse and human embryonic and induced pluripotent stem cells (ESCs and iPSCs). Single nucleotide variants, copy number variations, and larger chromosomal abnormalities may influence the differentiation capacity of pluripotent stem cells and the functionality of their derivatives in disease modeling and drug screening, and are considered a serious risk for cellular therapies based on ESC or iPSC derivatives. This review discusses the types and origins of different genetic abnormalities in pluripotent stem cells, methods for their detection, and the mechanisms of development and enrichment during reprogramming and culture expansion. [ABSTRACT FROM AUTHOR]

548 shark species were considered in this paper. They were classified in function of their depth of occurrence: 218 deep, 114 transitory and 210 shallow. The diets of the 332 deep and transitory species are reviewed. 10 prey categories are recognized here: Chondrichthyes, Teleosts, Cephalopods, Crustaceans, Marine mammals, Annelids, Ctenophores, Egg of Chondrichthyes, Siphonophores, and Bivalves. There is a complete lack of data for 210 species, and 59 have only a limited amount of information available. In general, teleosts, crustaceans, and cephalopods were the main prey found in deep-water sharks. However, some species have a specialized diet. A simple index of dietary knowledge has been developed to highlight the current state of knowledge on the subject, further illustrating the lack of information. The proportion of empty and regurgitated stomachs varies significantly between species and studies. Most data on deep and transitory sharks come from stomach content analysis. Stable isotope analysis provides additional insight into their trophic ecology. Fatty acid profiling and DNA metabarcoding can also add important information, but their use is currently limited with deep-water sharks. The most important conclusion from this synthesis is the lack or scarcity of information for most deep and transitory species. Graphical abstract (COLOR): Representation of the main prey taxa of four different families. A: Squalidae, B: Centrophoridae, C: Etmopteridae, D: Hexanchidae. The size of the prey taxa indicates its relative importance thourough the different studies reviewed. [Display omitted] • We reviewed the diet of deep-water and transitory shark species. • The species are classified in function of depth: 218 deep-water, 114 transitory, and 210 shallow. • In general, teleosts, crustaceans, and cephalopods were the main prey found in deep-water sharks. • A diet knowledge index is proposed, with most species lacking data on the diet with a score of 0. • Stable isotope analysis is more and more used, but fatty acid profiling and DNA metabarcoding are also interesting. [ABSTRACT FROM AUTHOR]

Constant mean curvature (CMC) tori in Euclidean 3-space are described by an algebraic curve, called the spectral curve, together with a line bundle on this curve and a point on S 1 , called the Sym point. For a given spectral curve the possible choices of line bundle and Sym point are easily described. The space of spectral curves of tori is totally disconnected. Hence to characterise the “moduli space” of CMC tori one should, for each genus g , determine the closure P g ¯ of spectral curves of CMC tori within the spectral curves of CMC planes having spectral genus g . We identify a real subvariety R g and a subset S g ⊆ R g such that R max g ⊆ P g ¯ ⊆ S g , where R max g denotes the points of R g having maximal dimension. The lowest spectral genus for which tori exist is g = 2 and in this case R 2 = R max 2 = P 2 ¯ = S 2 . For g > 2 , we conjecture that R g ⊋ R max g = S g . We give a number of alternative characterisations of R max g and in particular introduce a new integer invariant of a CMC plane of finite type, called its winding number. [ABSTRACT FROM AUTHOR]

304L stainless steel syntactic foam test specimens were produced by means of injection moulding of feedstock made from pre-alloyed 304L powder, binder and additions of S60HS micro hollow glass spheres and Fillite 106 cenospheres. After de-binding and sintering the foam pore structure and matrix micro-structure were analysed. Mechanical characterisation was done using tensile and compression tests. The results of the mechanical tests are discussed in terms of the observed foam and matrix microstructures. [ABSTRACT FROM AUTHOR]

Abstract: The induction of teratoma in mice by the transplantation of stem cells into extra-uterine sites has been used as a read-out for cellular pluripotency since the initial description of this phenomenon in 1954. Since then, the teratoma assay has remained the assay of choice to demonstrate pluripotency, gaining prominence during the recent hype surrounding human stem cell research. However, the scientific significance of the teratoma assay has been debated due to the fact that transplanted cells are exposed to a non-physiological environment. Since many mice are used for a result that is heavily questioned, it is time to reconsider the teratoma assay from an ethical point of view. Candidate alternatives to the teratoma assay comprise the directed differentiation of pluripotent stem cells into organotypic cells, differentiation of cells in embryoid bodies, the analysis of pluripotency-associated biomarkers with high correlation to the teratoma forming potential of stem cells, predictive epigenetic footprints, or a combination of these technologies. Each of these assays is capable of addressing one or more aspects of pluripotency, however it is essential that these assays are validated to provide an accepted robust, reproducible alternative. In particular, the rapidly expanding number of human induced pluripotent stem cell lines, requires the development of simple, affordable standardized in vitro and in silico assays to reduce the number of animal experiments performed. [Copyright &y& Elsevier]

Abstract: Therapeutic application of stem cell derivatives requires large quantities of cells produced in defined media that cannot be produced via conventional adherent culture. We have applied human induced pluripotent stem (hiPS) cells expressing eGFP under control of the OCT4 promoter to establish the expansion of undifferentiated human embryonic stem (hES) and hiPS cells in suspension culture. A defined culture medium has been identified that results in up to six-fold increase in cell numbers within four days. Our culture system is based on initial single cell dissociation which is critical for standardized process inoculation. HES / hiPS cells were expanded for up to 17 passages. The cells maintained a stable karyotype, their expression of pluripotency markers and their potential to differentiate into derivatives of all three germ layers. The ability to expand HES / hiPS cells in a scalable suspension culture represents a critical step towards standardized production in stirred bioreactors. [Copyright &y& Elsevier]

Medulloblastoma is the most common malignant brain tumor in childhood and adolescence. Although some patients present with distinct genetic alterations, such as mutated TP53 or MYC amplification, pediatric medulloblastoma is a tumor entity with minimal mutational load and low immunogenicity. We identified tumor-specific mutations using next-generation sequencing of medulloblastoma DNA and RNA derived from primary tumor samples from pediatric patients. Tumor-specific mutations were confirmed using deep sequencing and in silico analyses predicted high binding affinity of the neoantigen-derived peptides to the patients' human leukocyte antigen molecules. Tumor-specific peptides were synthesized and used to induce a de novo T-cell response characterized by interferon gamma and tumor necrosis factor alpha release of CD8+ cytotoxic T cells in vitro. Despite low mutational tumor burden, at least two immunogenic tumor-specific peptides were identified in each patient. T cells showed a balanced CD4/CD8 ratio and mostly effector memory phenotype. Induction of a CD8-specific T-cell response was achieved for the neoepitopes derived from Histidine Ammonia-Lyase (HAL), Neuraminidase 2 (NEU2), Proprotein Convertase Subtilisin (PCSK9), Programmed Cell Death 10 (PDCD10), Supervillin (SVIL) and tRNA Splicing Endonuclease Subunit 54 (TSEN54) variants. Detection of patient-specific, tumor-derived neoantigens confirms that even in tumors with low mutational load a molecular design of targets for specific T-cell immunotherapy is possible. The identified neoantigens may guide future approaches of adoptive T-cell transfer, transgenic T-cell receptor transfer or tumor vaccination. [ABSTRACT FROM AUTHOR]

Currently the ecology of deep-water sharks is poorly documented, especially in situ information for these elusive species are lacking. In this study, stereo-Baited Remote Underwater Videos (stereo-BRUVs) were deployed to collect ecological data from New Zealand deep-sea sharks. The results showed differences in abundance between species, with Etmopterus granulosus (Etmopteridae) found in greatest numbers. Moreover, the known depth range increased for Scymnodon macracanthus (Centrophiridae). Deep-sea shark species were generally found to swim at slower cruise speeds (0.36 ± 0.04 m s−1) than their shallow-water counterparts (0.63 ± 0.05 m s−1). However, the swimming speed of deep-sea sharks was clearly not uniform, with some species displaying higher cruise swimming speeds than others. The fastest sharks (Centrophorus harrissoni , Etmopterus granulosus and Etmopterus molleri) had swimming abilities comparable to benthic shallow water sharks (0.48 ± 0.02 m s−1). The higher cruise swimming speed in the family Etmopteridae could be an advantage for these luminous sharks if they follow isolumes to match their ventral light intensity with the down-welling light of their environment. This study revealed that alternative non-destructive methods can be effective for ecological studies of deep-sea marine fauna. • Deep-sea sharks display a lower cruise swimming speed than shallow-water ones. • Luminous Etmopteridae swim faster than others deep-sea species. • Different swimming speeds and tail beat frequencies in different deep-sea species. • The occurrence and relative abundance of sharks at different depths. • Slight or no size change with depth. [ABSTRACT FROM AUTHOR]

Computer-based analysis of long-term electrocardiogram (ECG) monitoring in animal models represents a cost and time-consuming process as manual supervision is often performed to ensure accuracy in arrhythmia detection. Here, we investigate the performance and feasibility of three ECG interval analysis approaches A) attribute-based, B) attribute- and pattern recognition-based and C) combined approach with additional manual beat-to-beat analysis (gold standard) with regard to subsequent detection of ventricular arrhythmias (VA) and time consumption. ECG analysis was performed on ECG raw data of 5 male cynomolgus monkeys (1000 h total, 2 × 100 h per animal). Both approaches A and B overestimated the total number of arrhythmias compared to gold standard (+8.92% vs. +6.47%). With regard to correct classification of detected VA event numbers (accelerated idioventricular rhythms [AIVR], ventricular tachycardia [VT]) approach B revealed higher accuracy compared to approach A. Importantly, VA burden (% of time) was precisely depicted when using approach B (−1.13%), whereas approach A resulted in relevant undersensing of ventricular arrhythmias (−11.76%). Of note, approach A and B could be performed with significant less working time (−95% and − 91% working time) compared to gold standard. In sum, we show that a combination of attribute-based and pattern recognition analysis (approach B) can reproduce VA burden with acceptable accuracy without using manual supervision. Since this approach allowed analyses to be performed with distinct time saving it represents a valuable approach for cost and time efficient analysis of large preclinical ECG datasets. [ABSTRACT FROM AUTHOR]

Deep-sea elasmobranchs are commonly reported as bycatch of deep-sea fisheries and their subsequent loss has been highlighted as a long-running concern to the ecosystem ecological functioning. To understand the possible consequences of their removal, information on basic ecological traits, such as diet and foraging strategies, is needed. Such aspects have been widely studied through stomach content analysis but the lack of long-term dietary information requires other tools to be used such as stable isotopes. This study examines nitrogen and carbon isotope compositions of the velvet belly lantern shark, Etmopterus spinax , one of the most impacted shark species in northeastern Atlantic fisheries as a result of accidental catches. E. spinax was sampled at four different locations, characterized by contrasting oceanographic and ecological conditions: the western Mediterranean Sea (near the Balearic Islands), the southern Iberian upwelling system, Rockall Trough and southwestern Norwegian fjords. Stomach content analysis revealed similar prey species among sites, with a diet dominated by Euphausiacea (mostly Meganyctiphanes norvegica) and an ontogenetic shift towards small teleost fishes, cephalopods or other crustaceans. Despite these similarities, muscle stable isotope compositions differed across sampled locations. Rather than clear dietary differences, the contrasted isotopic values are likely to reflect differences in environmental settings and biogeochemical processes affecting nutrient dynamics at the base of the food webs. • E. spinax were sampled across the northeastern Atlantic and Mediterranean Sea. • Stomach contents suggested a similar diet. • E. spinax fed mainly on Euphausiacea with an ontogenetic diet shift. • Muscle carbon and nitrogen isotope compositions differed between locations. • Isotopic differences probably resulted from mechanisms affecting the baselines. [ABSTRACT FROM AUTHOR]

Abstract: In most pluripotent stem cell differentiation protocols the formation of embryoid bodies (EBs) is an important step. Here we describe a rapid, straightforward soft lithography approach for the preparation of hydrophilic silicon masters from different templates and the subsequent production of patterned agarose-DMEM microwell surfaces for scalable well standardized stem cell aggregation and EB formation. The non-adhesive agarose microwell plates represent an accurate replication of the templates'' topography and were used for aggregation of murine induced pluripotent stem cells (iPSCs) and human embryonic stem cells (ESCs). Direct microscopic assessment by time-lapse analysis demonstrated rapid formation of uniformly shaped EBs from murine iPSCs with similar or even more consistent results concerning size distribution and harvesting efficiency compared to the commonly used but time-consuming hanging drop technique. For human ESCs, homogenous aggregates were obtained after single cell inoculation on agarose microwells with efficient differentiation into the cardiac lineage using state-of-the-art protocols for directed differentiation via small molecules. With this soft lithography-based strategy, sufficient and reproducible numbers of stem cell-derived cardiomyocytes necessary for tissue engineering purposes can be realized in a highly controllable manner. Moreover, it might be useful for different cell types in any application that requires scalable and highly standardized aggregation. [Copyright &y& Elsevier]

Abstract: Despite recent major advances including reprogramming and directed cardiac differentiation of human cells, therapeutic application of in vitro engineered myocardial tissue is still not feasible due to the inability to construct functional large vascularized contractile tissue patches based on clinically applicable and fully defined matrix components. Typical matrices with preformed porous 3D structure cannot be applied due to the obvious lack of migratory capacity of cardiomyocytes (CM). We have therefore developed a fully defined in situ hydrogelation system based on alginate (Alg) and hyaluronic acid (HyA), in which their aldehyde and hydrazide-derivatives enable covalent hydrazone cross-linking of polysaccharides in the presence of viable myocytes. By varying degrees of derivatization, concentrations and composition of blends in a modular system, mechanophysical properties of the resulting hydrogels are easily adjustable. The hydrogel allowed for the generation of contractile bioartificial cardiac tissue from CM-enriched neonatal rat heart cells, which resembles native myocardium. A combination of HyA and highly purified human collagen I led to significantly increased active contraction force compared to collagen, only. Therefore, our in situ cross-linking hydrogels represent a valuable toolbox for the fine-tuning of engineered cardiac tissue''s mechanical properties and improved functionality, facilitating clinical translation toward therapeutic heart muscle reconstruction. [Copyright &y& Elsevier]