Adhesion Secretion Research Articles

Mesocosm and Microcosm Experiments On the Feeding of Temperate Salt Marsh Foraminifera

AbstractAgglutinated foraminifera dominate in temperate salt marsh sediment, making them key indicators for monitoring sea level and environmental changes. Little is known about the biology of these benthic foraminifera because of difficulties in distinguishing live from dead specimens in laboratory cultures. We present data from 10 years of laboratory experiments using comparisons of the agglutinant trochamminids Trochammina inflata and Entzia macrescens and the miliolid Miliammina fusca with the calcareous rotalids Helenina anderseni and Elphidium williamsoni. Specimens were taken from a laboratory mesocosm representing Chezzetcook Inlet, a cool-temperate salt marsh in eastern Canada. We determined culture requirements for the agglutinated foraminifera in Petri dishes over 10–12 week periods. Five inexpensive, non-terminal ways of identifying live organisms were developed: spatial movement, detritus-gathering, attachment, clustering, and test opacity. Comparison with rose Bengal staining showed <10% diversion for calcareous species and T. inflata but M. fusca was over-counted by >30%. Terminal chambers of Trochammina inflata were examined by transmission electron microscopy to visualise food consumption and identify food in digestive vacuoles, both in specimens from mesocosm and in culture. Bacteria and unidentified detritus in the vacuoles establish that this agglutinated species is a saprophagous and bacterivorous detritivore. The adhesive secretions by these species apparently help them gather and possibly farm food while being relatively immobile in the sediments. Our observations of movement and feeding orientation in the agglutinants suggest links between form and function that underscore their value as ultra high resolution sea-level proxies. Mesocosm biomass and abundance counts show that foraminifera represent >50% of the meiofaunal biomass, emphasising their importance in the food web and energy-flow dynamics of temperate salt marsh systems.

Mechanisms utilized by feline adipose-derived mesenchymal stem cells to inhibit T lymphocyte proliferation

BackgroundFeline adipose-derived mesenchymal stem cells (ASCs) have been successfully used in clinical trials for the treatment of immune-mediated diseases with T cell dysregulation. However, the immunomodulatory pathways utilized by feline ASCs to suppress T cell activation have not been fully determined. We investigated the mechanisms used by feline ASCs to inhibit T cell proliferation, including the soluble factors and the cell-cell contact ligands responsible for ASC-T cell interaction.MethodsThe immunomodulatory activity of feline ASCs was evaluated via cell cycle analysis and in vitro mixed leukocyte reaction using specific immunomodulatory inhibitors. Cell-cell interactions were assessed with static adhesion assays, also with inhibitors.ResultsFeline ASCs decrease T cell proliferation by causing cell cycle arrest in G0–G1. Blocking prostaglandin (PGE2), but not IDO, partially restored lymphocyte proliferation. Although PDL-1 and CD137L are both expressed on activated feline ASCs, only the interaction of intercellular adhesion molecule 1 (ICAM-1, CD54) with its ligand, lymphocyte function-associated antigen 1 (LFA-1, CD11a/CD18), was responsible for ASC-T cell adhesion. Blocking this interaction reduced cell-cell adhesion and mediator (IFN-γ) secretion and signaling.ConclusionsFeline ASCs utilize PGE2 and ICAM-1/LFA-1 ligand interaction to inhibit T cell proliferation with a resultant cell cycle arrest in G0–G1. These data further elucidate the mechanisms by which feline ASCs interact with T cells, help define appropriate T cell-mediated disease targets in cats that may be amenable to ASC therapy, and may also inform potential translational models for human diseases.

Adhesive defence mucus secretions in the red triangle slug (Triboniophorus graeffei) can incapacitate adult frogs

AbstractGastropods are known to secrete mucus for a variety of purposes, including locomotion, reproduction, adhesion to surfaces and lubrication. A less commonly known function of mucus secretion in this group involves its use as a defence against predation. Amongst the terrestrial slugs, mucus that serves this particular purpose has been studied for only a handful of species under laboratory conditions, where it is thought to be produced to make individuals unpalatable or difficult to consume. However, the mechanisms of how these defensive secretions operate and their effectiveness in deterring predation in the natural world have not been described in much detail. In this study, we provide evidence of adhesive mucus secretions in the red triangle slug (Triboniophorus graeffei) as an adaptation against predation. Field observations of a large red‐eyed green tree frog (Litoria chloris) trapped in the mucus secretions of a nearby T. graeffei revealed that this mucus serves to incapacitate predators rather than just an overall deterrence. Mechanical stimulation of T. graeffei under laboratory conditions revealed that adhesive secretions were produced from discrete sections of the dorsal surface when disturbed, leading to the production of a highly sticky and elastic mucus that was unlike the thin and slippery mucus used during locomotion. The adhesiveness of the defensive secretions was strengthened and reactivated when in contact with water. This appears to not only be the first description of defensive mucus production in this slug species but one of the first natural observations of slug secretions incapacitating a relatively large vertebrate. The biomechanical properties of this mucus and its ability to maintain and strengthen its hold under wet conditions make it potentially useful in the development of new adhesive materials.

Silk fibroin coating through EDC/NHS crosslink is an effective method to promote graft remodeling of a polyethylene terephthalate artificial ligament.

Anterior cruciate ligament reconstruction using polyethylene terephthalate artificial ligaments is one of the research hotspots in sports medicine but it is still challenging to achieve biological healing. The purpose of this study was to modify polyethylene terephthalate ligament with silk fibroin through ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)/N-hydroxysuccinimide (NHS) crosslink and to investigate the performance of graft remodeling in vitro and in vivo. After silk fibroin coating, changes in the surface properties of ligament were characterized by scanning electron microscopy, attenuated total reflectance-Fourier transform infrared spectroscopy and water contact angle measurements. The compatibility of polyethylene terephthalate ligament with silk fibroin coating was investigated in vitro. The results showed the silk fibroin coating significantly improved adhesion, proliferation and extracellular matrix secretion of fibroblast cells. Moreover, a rabbit anterior cruciate ligament reconstruction model was established to evaluate the effect of ligament with silk fibroin coating in vivo. The gross observation and histological results showed that the silk fibroin coating significantly inhibited inflammation response and promoted new tissue regeneration with fusiform cells infiltration in and around the graft. Furthermore, the expressions of collagen I protein and mRNA in the silk fibroin-coated polyethylene terephthalate group were much higher than those in the control group according to the immunohistochemical and real-time polymerase chain reaction results. Therefore, silk fibroin coating through EDC/NHS crosslink promotes the biocompatibility and remodeling process of polyethylene terephthalate artificial ligament in vitro and in vivo. It can be considered as a potential solution to the problem of poor remodeling of artificial ligaments after anterior cruciate ligament reconstruction in the clinical applications.

Ultrastructure of the anterior adhesive apparatus of the gill parasite Macrogyrodactylus clarii and skin parasite M. congolensis (Monogenea; Gyrodactylidae) from the catfish Clarias gariepinus

Transmission electron microscopy (TEM) was used for the first time to study the anterior adhesive apparatus of the monogeneans Macrogyrodactylus clarii Gussev, 1961 and M. congolensis (Prudhoe, 1957) Yamaguti, 1963 inhabiting gills and skin respectively of the same catfish Clarias gariepinus. Despite the different microhabitats occupied by these parasites, the present study revealed that they have a similar anterior adhesive system. In both parasites, the anterior adhesive apparatus consists of three types of gland cells: G1 cells that produce rod-shaped bodies (S1), G2 cells manufacture irregularly shaped bodies (S2) and G3 cells form mucoid-like secretions (S3). In the cytoplasm of G1 cells, a single layer of microtubules encloses each developing rod-shaped body. A unique feature of S1 secretory bodies is that some fully developed S1 bodies are attached to each other, forming large condensed globules in the cytoplasm of G1 gland cells and terminal portion of the G1 ducts, but none were detected in the adhesive sacs outside the ducts. In the adhesive sacs, G1 ducts open with multiple apertures whereas each of the G2 and G3 ducts have a single opening. The adhesive sacs are lined with two types of tegument (st1 and st2). A third tegument type (st3) connects the st2 tegument with the general body tegument. Only st1 has microvilli. Each adhesive sac is provided with a spike-like sensillum and single uniciliated sense organ. The possible functions of microvilli in increasing the surface area and assistance in spreading and mixing of the adhesive secretion, and the role of sense organs associated with the adhesive sacs are discussed.

Biomechanical properties of fishing lines of the glowworm Arachnocampa luminosa (Diptera; Keroplatidae)

Animals use adhesive secretions in highly diverse ways, such as for settlement, egg anchorage, mating, active or passive defence, etc. One of the most interesting functions is the use of bioadhesives to capture prey, as the bonding has to be performed within milliseconds and often under unfavourable conditions. While much is understood about the adhesive and biomechanical properties of the threads of other hunters such as spiders, barely anything is documented about those of the New Zealand glowworm Arachnocampa luminosa. We analysed tensile properties of the fishing lines of the New Zealand glowworm Arachnocampa luminosa under natural and dry conditions and measured their adhesion energy to different surfaces. The capture system of A. luminosa is highly adapted to the prevailing conditions (13–15 °C, relative humidity of 98%) whereby the wet fishing lines only show a bonding ability at high relative humidity (>80%) with a mean adhesive energy from 20–45 N/m and a stronger adhesion to polar surfaces. Wet threads show a slightly higher breaking strain value than dried threads, whereas the tensile strength of wet threads was much lower. The analyses show that breaking stress and strain values in Arachnocampa luminosa were very low in comparison to related Arachnocampa species and spider silk threads but exhibit much higher adhesion energy values. While the mechanical differences between the threads of various Arachnocampa species might be consequence of the different sampling and handling of the threads prior to the tests, differences to spiders could be explained by habitat differences and differences in the material ultrastructure. Orb web spiders produce viscid silk consisting of β-pleated sheets, whereas Arachnocampa has cross-β–sheet crystallites within its silk. As a functional explanation, the low tear strength for A. luminosa comprises a safety mechanism and ensures the entire nest is not pulled down by prey which is too heavy.

Secretome analysis of Strongyloides venezuelensis parasitic stages reveals that soluble and insoluble proteins are involved in its parasitism

BackgroundParasites excrete and secrete a wide range of molecules that act as the primary interface with their hosts and play critical roles in establishing parasitism during different stages of infection. Strongyloides venezuelensis is a gastrointestinal parasite of rats that is widely used as a laboratory model and is known to produce both soluble and insoluble (adhesive) secretions during its parasitic stages. However, little is known about the constituents of these secretions.ResultsUsing mass spectrometry, we identified 436 proteins from the infective third-stage larvae (iL3s) and 196 proteins from the parasitic females of S. venezuelensis. The proteins that were secreted by the iL3s were enriched with peptidase activity, embryo development and the oxidation-reduction process, while those of the parasitic females were associated with glycolysis, DNA binding (histones) and other unknown functions. Trypsin inhibitor-like domain-containing proteins were identified as the main component of the adhesive secretion from parasitic females. An absence of secretion signals in many of the proteins indicated that they are secreted via non-classical secretion pathways.ConclusionsWe found that S. venezuelensis secretes a wide range of proteins to establish parasitism. This includes proteins that have previously been identified as being involved in parasitism in other helminths as well as proteins that are unique to this species. These findings provide insights into the molecular mechanisms underlying Strongyloides parasitism.

Silk–Graphene Hybrid Hydrogels with Multiple Cues to Induce Nerve Cell Behavior

Cell behavior is dependent in part on chemical and physical cues from the extracellular matrix. Although the influence of various cues on cell behavior has been studied, challenges remain to incorporate multiple cues to matrix systems to optimize and control cell outcomes. Here, aligned silk fibroin (SF)-graphene hydrogels with preferable stiffness were developed through arranging SF nanofibers and SF-modified graphene sheets under an electric field. Different signals, such as bioactive graphene, nanofibrous structure, aligned topography, and mechanical stiffness, were tailored into the hydrogel system, providing niches for nerve cell responses. The desired adhesion, proliferation, differentiation, extensio,n and growth factor secretion of multiple nerve-related cells was achieved on these hydrogels, suggesting strong synergistic action through the combination of different cues. Based on the fabrication strategy, our present study provides a useful materials engineering platform for revealing cooperative influences of different signals on nerve cell behavior, to help in the understanding of cell-biomaterial interactions, with potential toward studies related to nerve regeneration.

Molecular Expression of Bone Marrow Angiogenic Factors, Cell-Cell Adhesion Molecules and Matrix-Metallo-Proteinases in Plasmacellular Disorders: a Molecular Panel to İnvestigate Disease Progression.

Increasing levels of angiogenesis play an important role in the pathogenesis and progression of multiple myeloma (MM). Malignant plasma cells promote a gradual increase in the degree of angiogenesis, modulation of specific cell-cell adhesion molecules and secretion of matrix-metallo-proteinases (MMPs), changing the BM composition from benign conditions, such as MGUS, to smouldering multiple myeloma (SM) and to active MM. We aimed to identify a gene expression profile, helpful to discriminate the “angiogenic potential” in BM and PB plasma cells from MGUS, SMM and active MM patients analyzed at diagnosis. We analyzed the expression of cell-cell adhesion molecules such as VE-Cadherin, E-Cadherin MCAM/MUC18/CD146 and of the MMP-2 and MMP-9. MCAM/MUC18 expression resulted mostly associated with that of the pivotal angiogenic factors VEGF and Ang2, and in MGUS the pattern was different in steady state, compared to progression towards SM. Furthermore, E-Cadherin, the main epithelial cell-cell-adhesion molecule, unexpectedly resulted overexpressed in MM.
 

A Novel Approach via Surface Modification of Degradable Polymers With Adhesive DOPA-IGF-1 for Neural Tissue Engineering

The highly damaging state of spinal cord injuries has provided much inspiration for the design of surface modification of the implants that can promote nerve regeneration and functional reconstruction. DOPA-IGF-1, a new recombinant protein designed in our previous study, exhibited strong binding affinity to titanium and significantly enhanced the growth of NIH3T3 cells on the surface of titanium with the same biological activity as IGF-1. In this article, surface modification of poly(lactide-co-glycolide) (PLGA) films with recombinant DOPA-IGF-1 was performed to promote the paracrine activity of human umbilical cord mesenchymal stem cells (hUCMSCs) by secreting neurotrophic factors. DOPA-IGF-1 exhibited the strongest binding ability to PLGA films than commercial IGF-1 and nonhydroxylated YKYKY-IGF-1. In vitro cultures of hUCMSCs on the modified PLGA films showed that DOPA-IGF-1@PLGA substrates significantly improved the proliferation, adhesion, and neurotrophic factors secretion of hUCMSCs, especially for nerve growth factor, as confirmed by qRT-PCR and western blot analysis. Subsequently, the acquired neurotrophic factors secreted by the hUCMSCs cultured on the DOPA-IGF-1@PLGA films obviously enhanced neurite outgrowth of PC12 cells. Taken together, PLGA substrates with DOPA-IGF-1 immobilization is a promising platform for neural tissue engineering via neurotrophic factors secretion from MSCs and should be further tested in vivo.

Properties of temporary adhesion systems of marine and freshwater organisms.

Underwater adhesive secretions are a promising source of inspiration for biomedical and industrial applications. Although marine permanent adhesives have been extensively investigated, reversible adhesion, e.g. as used for locomotion and feeding, is still poorly understood. Here, we summarise the current knowledge on secretion-based, temporary adhesive systems in aquatic environments, with a special emphasis on the morphology and structure of adhesive organs and adhesive material. Many animals employing temporary adhesion to the substratum rely on so-called duo-gland adhesive organs, consisting of two secretory gland cells and one supportive cell. We give a detailed depiction of a basic duo-gland adhesive organ and variations thereof. Additionally, we discuss temporary adhesive systems with an alternative building plan. Next, the topography of secreted adhesive footprints is described based on examples. The limited data on the composition of temporary adhesives are summarised, separating known protein components and carbohydrate residues. There are still large gaps in our understanding of temporary adhesion. We discuss three proposed models for detachment, although the actual mechanism of voluntary detachment is still a matter for debate.

Transcriptome analysis of Porphyromonas gingivalis and Acinetobacter baumannii in polymicrobial communities.

Acinetobacter baumannii is a nosocomial, opportunistic pathogen that causes several serious conditions including meningitis, septicemia, endocarditis, and pneumonia. It can be found in the oral biofilm, which may be a reservoir for pneumonia and chronic obstructive pulmonary disease. Subgingival colonization by A.baumannii is associated with chronic and aggressive periodontitis as well as refractory periodontal disease. Porphyromonas gingivalis, a keystone periodontal pathogen localized to subgingival plaque, is also implicated in several chronic conditions including aspiration pneumonia. Although both bacteria are found together in subgingival plaque and can cause multiple polymicrobial infections, nothing is known about the interactions between these two important human pathogens. In this study, we used RNA sequencing to understand the transcriptional response of both species as they adapt to heterotypic communities. Among the differentially regulated genes were those encoding a number of important virulence factors for both species including adhesion, biofilm formation, and protein secretion. Additionally, the presence of A.baumannii increased the abundance of P.gingivalis in model dual-species communities. Collectively these results suggest that both P.gingivalis and A.baumannii adapt to each other and have synergistic potential for increased pathogenicity. In identifying the mechanisms that promote pathogenicity and refractory disease, novel approaches to mitigate polymicrobial synergistic interactions may be developed to treat or prevent associated diseases.

Genome-wide analysis of circular RNAs in bovine cumulus cells treated with BMP15 and GDF9

Circular RNAs (circRNAs) are important members of the non-coding RNA family, and those relating to animal physiologies have been widely studied in recent years. This study aimed to explore the roles of circRNAs in the regulation of follicular development. We constructed four bovine cumulus cell cDNA libraries, including a negative control group (NC) and groups treated with BMP15, GDF9 and BMP15 + GDF9, and we sequenced the libraries on the Illumina HiSeq Xten platform. We identified 1706 circRNAs and screened for differential circRNA expression. We conducted a bioinformatics analysis of these circRNAs and screened for differential circRNAs. Functional annotation and enrichment analysis of the host genes showed that the differential circRNAs were related to locomotion, reproduction, biological adhesion, growth, rhythmic processes, biological phases and hormone secretion. According to the differential expression of circRNA between groups, there were 3 up-regulated and 6 down-regulated circRNAs in the BMP15 group as well as 12 up-regulated and 24 down-regulated circRNAs in the GDF9 group. Co-addition of both BMP15 and GDF9 resulted in 15 up-regulated and 13 down-regulated circRNAs. circ_n/a_75,circ_12691_1 and circ_n/a_303 were altered in both the BMP15 and GDF9 groups as well as in the BMP15 + GDF9 combination group. We focused on these three circRNAs because they were potentially associated with the additive effect of BMP15 and GDF9. Quantitative PCR analysis showed that the expression levels of these three circRNAs were consistent with the sequencing results. In addition, the target miRNAs of circ_n/a_75 and circ_n/a_303, miR-339a, miR-2400 and miR-30c, were down-regulated in the experimental group, which was in contrast to the circRNAs trend. These findings demonstrated that BMP15 and GDF9 may regulate the target gene through circRNA, as a miRNA sponge, in order to regulate the status of bovine cumulus cells and affect follicular development.

A retinoic acid-dependent stroma-leukemia crosstalk promotes chronic lymphocytic leukemia progression

In chronic lymphocytic leukemia (CLL), the non-hematopoietic stromal microenvironment plays a critical role in promoting tumor cell recruitment, activation, survival, and expansion. However, the nature of the stromal cells and molecular pathways involved remain largely unknown. Here, we demonstrate that leukemic B lymphocytes induce the activation of retinoid acid synthesis and signaling in the microenvironment. Inhibition of RA-signaling in stromal cells causes deregulation of genes associated with adhesion, tissue organization and chemokine secretion including the B-cell chemokine CXCL13. Notably, reducing retinoic acid precursors from the diet or inhibiting RA-signaling through retinoid-antagonist therapy prolong survival by preventing dissemination of leukemia cells into lymphoid tissues. Furthermore, mouse and human leukemia cells could be distinguished from normal B-cells by their increased expression of Rarγ2 and RXRα, respectively. These findings establish a role for retinoids in murine CLL pathogenesis, and provide new therapeutic strategies to target the microenvironment and to control disease progression.

Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion.

Chemerin, a chemoattractant protein, acts via a G-protein coupled chemokine receptor, i.e. Chemokine like Receptor 1/ChemR23; levels of which are elevated in pro-inflammatory states such as obesity and type 2 diabetes mellitus (T2DM). Obesity and T2DM patients are at high risk of developing cardiovascular disorders such as atherosclerosis. We have reported that chemerin induces human endothelial cell angiogenesis and since dysregulated angiogenesis and endothelial dysfunction are hallmarks of vascular disease; we sought to determine the effects of chemerin on monocyte-endothelial adhesion, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), a critical pro-inflammatory transcription factor. Human endothelial cells were transfected with pNF-kappaB-Luc plasmid. Chemerin induced NF-κB activation via the MAPK and PI3K/Akt pathways. Western blot analyses and monocyte-endothelial adhesion assay showed that chemerin increased endothelial cell adhesion molecule expression and secretion, namely E-selectin (Endothelial Selectin), VCAM-1 (Vascular Cell Adhesion Molecule-1) and ICAM-1 (Intracellular Adhesion Molecule-1), leading to enhancement of monocyte-endothelial adhesion. Additionally, we showed a synergistic response of the pro-inflammatory mediator, Interleukin-1β with chemerin induced effects. Chemerin plays an important role in endothelial inflammation, as it induces monocyte-endothelial adhesion, a critical step in the development of atherosclerosis.

Ordered Topographically Patterned Silicon by Insect-Inspired Capillary Submicron Stamping.

Insect-inspired capillary submicron stamping and subsequent surface-limited metal-assisted chemical etching (MACE) with ammonium bifluoride as a HF source are employed for the high-throughput production of ordered topographically patterned silicon (tpSi). Insect feet often possess hairy contact elements through which adhesive secretion is deployed. Thus, arrays of adhesive secretion drops remain as footprints on contact surfaces. Stamps for insect-inspired capillary submicron stamping having surfaces topographically patterned with contact elements mimic the functional principles of such insect feet. They contain spongy continuous nanopore networks penetrating the entire stamps. Any ink (organic or aqueous) may be supplied from the backside of the nanoporous stamps to the contact elements. We generated ordered arrays of submicron AgNO3 dots extending square millimeters on Si by manual stamping with cycle times of a few seconds under ambient conditions; at higher load, ordered holey AgNO3 films were obtained. Surface-limited MACE correspondingly yielded either macroporous tpSi or Si pillar arrays. Inkjet printing of polymer solutions onto the tpSi yielded patterns of polymer blots conformally covering the tpSi. Such blot patterns could potentially represent a starting point for the development of persistent and scratch-resistant identity labels or quick response codes on silicon surfaces.

Involvement of sulfated biopolymers in adhesive secretions produced by marine invertebrates.

ABSTRACTMany marine invertebrates use adhesive secretions to attach to underwater surfaces and functional groups borne by their adhesive proteins and carbohydrates, such as catechols and phosphates, play a key role in adhesion. The occurrence of sulfates as recurrent moieties in marine bioadhesives suggests that they could also be involved. However, in most cases, their presence in the adhesive material remains speculative. We investigated the presence of sulfated biopolymers in five marine invertebrates representative of the four types of adhesion encountered in the sea: mussels and tubeworms for permanent adhesion, limpets for transitory adhesion, sea stars for temporary adhesion and sea cucumbers for instantaneous adhesion. The dry adhesive material of mussels, sea stars and sea cucumbers contained about 1% of sulfate. Using anti-sulfotyrosine antibodies and Alcian Blue staining, sulfated proteins and sulfated proteoglycans and/or polysaccharides were identified in the secretory cells and adhesive secretions of all species except the tubeworm. Sulfated proteoglycans appear to play a role only in the non-permanent adhesion of sea stars and limpets in which they could mediate cohesion within the adhesive material. In mussels and sea cucumbers, sulfated biopolymers would rather have an anti-adhesive function, precluding self-adhesion.

Electron Tomography and Correlative Approaches in Platelet Studies.

Blood platelets play a central role in the arrest of bleeding and the development of thrombosis. Unraveling the complex processes of platelet biogenesis from megakaryocytes, platelet adhesion, aggregation, and secretory responses are important topics in the field of hemostasis and thrombosis. Analysis of the ultrastructural changes that occur during these processes is essential for understanding the rapid membrane dynamics and has contributed substantially to our present knowledge of platelet formation and functioning. Recent developments in real-time imaging, correlative light and electron microscopy imaging (CLEM), and 3D (cryo) electron microscopy and tomography offer exciting opportunities to improve studies of the platelet adhesive responses and secretion at the ultrastructural level in a close to native environment. In this chapter we discuss and illustrate cryo preparation techniques (high-pressure freezing, vitrification), correlative LM and EM workflows, and 3D cryo-electron tomography that we apply in our current research projects.

Targetome Analysis Revealed Involvement of MiR-126 in Neurotrophin Signaling Pathway: A Possible Role in Prevention of Glioma Development.

Objective For the first time, we used molecular signaling pathway enrichment analysis to determine possible involvement of miR-126 and IRS-1 in neurotrophin pathway. Materials and Methods In this prospective study, validated and predicted targets (targetome) of miR-126 were collected following searching miRtarbase (http://mirtarbase.mbc.nctu.edu.tw/) and miRWalk 2.0 databases, respectively. Then, approximate expression of miR-126 targeting in Glioma tissue was examined using UniGene database (http://www.ncbi. nlm.nih.gov/unigene). In silico molecular pathway enrichment analysis was carried out by DAVID 6.7 database (http://david. abcc.ncifcrf.gov/) to explore which signaling pathway is related to miR-126 targeting and how miR-126 attributes to glioma development. Results MiR-126 exerts a variety of functions in cancer pathogenesis via suppression of expression of target gene including PI3K, KRAS, EGFL7, IRS-1 and VEGF. Our bioinformatic studies implementing DAVID database, showed the involvement of miR-126 target genes in several signaling pathways including cancer pathogenesis, neurotrophin functions, Glioma formation, insulin function, focal adhesion production, chemokine synthesis and secretion and regulation of the actin cytoskeleton. ConclusionTaken together, we concluded that miR-126 enhances the formation of glioma cancer stem cell probably via down regulation of IRS-1 in neurotrophin signaling pathway.

Extracellular micro and nanostructures forming the velvet worm solidified adhesive secretion

The onychophoran Epiperipatus hilkae secrets a sticky slime that solidifies almost immediately upon contact with air and under high humidy environmental condition forming a glassy like material. The general adhesive biochemical composition, the releasing and hardening mechanism have been partially described in literature. In this study, the structural characterization of the extracellular microstructures and nanostructures forming the solid adhesive of the secretion from Epiperipatus hilkae velvet worm is presented. The adhesive secretion is formed by macro-threads, which, in their solid state, are composed of globular particles approximately 700 nm in diameter that are distributed homogeneously throughout the matrix surface, and nanoparticles approximately 70 nm in diameter that and 6 nm in height self-assemble forming fiber-like structures. Nanoparticules with approximately 2 nm heights and others with non roundish forms are also observed. These 70 nm nano particles could be associated to proteins that form high density coverage films with low roughness; suggesting the formation of 2D ordered films. A crystalline and an amorphous phase composes the solidified secretion. The glassy or viscoelastic properties depend on the time in contact with air before being adhered to a solid surface and/or the mechanical stimulus; suggesting a key role of the drying on the hardening process.