Visualization of Brain Tumors with Infrared-Labeled Aptamers for Fluorescence-Guided Surgery.

Gliomas remain challenging brain tumors to treat due to their infiltrative nature. Accurately identifying tumor boundaries during surgery is crucial for successful resection. This study introduces an innovative intraoperative visualization method utilizing surgical fluorescence microscopy to precisely locate tumor cell dissemination.

Littoporins: Novel actinoporin-like proteins in caenogastropod genus Littorina.

In the course of searching for genes controlling the immune system in caenogastropod mollusks, we characterized and phylogenetically placed five new actinoporin-like cytolysins expressed in periwinkles of the genus Littorina. These newly discovered proteins, named littoporins (LitP), contain a central cytolysin/lectin domain and exhibit a predicted protein fold that is almost identical to the three-dimensional structures of actinoporins. Two of these proteins, LitP-1 and LitP-2, were found to be upregulated in L.

Neuronal gene expression in two generations of the marine parasitic worm, Cryptocotyle lingua.

Trematodes, or flukes, undergo intricate anatomical and behavioral transformations during their life cycle, yet the functional changes in their nervous system remain poorly understood. We investigated the molecular basis of nervous system function in Cryptocotyle lingua, a species of relevance for fisheries. Transcriptomic analysis revealed a streamlined molecular toolkit with the absence of key signaling pathways and ion channels.

Development of DNA aptamers for visualization of glial brain tumors and detection of circulating tumor cells.

Here, we present DNA aptamers capable of specific binding to glial tumor cells , , and for visualization diagnostics of central nervous system tumors. We selected the aptamers binding specifically to the postoperative human glial primary tumors and not to the healthy brain cells and meningioma, using a modified process of systematic evolution of ligands by exponential enrichment to cells; sequenced and analyzed ssDNA pools using bioinformatic tools and identified the best aptamers by their binding abilities; determined three-dimensional structures of lead aptamers (Gli-55 and Gli-233) with small-angle X-ray scattering and molecular modeling; isolated and identified molecular target proteins of the aptamers by mass spectrometry; the potential binding sites of Gli-233 to the target protein and the role of post-translational modifications were verified by molecular dynamics simulations. The anti-glioma aptamers Gli-233 and Gli-55 were used to detect circulating tumor cells in liquid biopsies.

Transcriptomic profiling of immune-associated molecules in the coelomocytes of lugworm Arenicola marina (Linnaeus, 1758).

Organization and functioning of immune system remain unevenly studied in different taxa of lophotrochozoan animals. We analyzed transcriptomic data on coelomocytes of the lugworm Arenicola marina (Linnaeus, 1758; Annelida, Polychaeta) to gain insights into the molecular mechanisms involved in polychaete immunity. Coelomocytes are specialized motile cells populating coelomic fluid of annelids, responsible for cellular defense reactions and providing humoral immune factors.

Identification of peptidoglycan recognition proteins in hemocytes and kidney of common periwinkle Littorinalittorea.

Peptidoglycan Recognition Proteins (PGRPs) are a diverse group of proteins involved in innate immunity. In particular, PGRPs have been shown to participate in immune pattern recognition in various mollusks. However, they have not been described in Caenogastropoda, a large molluscan group comprising sea, freshwater and land snails.

Toll-like signaling pathway in the transcriptome of Littorina littorea.

The transcriptome of the caenogastropod mollusk Littorina littorea was scanned for the presence of sequences encoding Toll-like receptors (TLRs) and corresponding proteins involved in downstream TLR signaling pathway. In the transcriptomic snapshots of hemocytes and kidney tissues, 45 complete TLRs encoded by 35 genes were identified. Out of the 59 non-TLR molecules involved in a canonical TLR signaling pathway, 35 genes were classified as homologous and could be placed within the TLR-mediated MyD88-and MAPK-dependent circuitries.

Immune response of a caenogastropod host: A case study of Littorina littorea and its digenean parasites.

The caenogastropod mollusk Littorina littorea is a promising experimental model for comparative studies on host/parasite immune conflict. Several different digenean parasites use L. littorea as the first intermediate host, overcoming snail immune reactions by a wide range of tactics that are radically different among different digenean species and at different developmental parasite stages.

Derivatives of the lectin complement pathway in Lophotrochozoa.

A plethora of non-overlapping immune molecular mechanisms in metazoans is the most puzzling issue in comparative immunobiology. No valid evolutionary retrospective on these mechanisms has been developed. In this study, we aimed to reveal the origin and evolution of the immune complement-like system in Lophotrochozoa.

Immune repertoire in the transcriptome of Littorina littorea reveals new trends in lophotrochozoan proto-complement evolution.

The evolution of complement system in invertebrates is poorly investigated. While the repertoire of complement genes in several Ecdysozoa lineages is found substantially different from that of Deuterostomia, the composition and function of the complement in the second protostome lineage, Lophotrochozoa, remains unclear. Here we report the general description of new transcriptomic data on the common periwinkle, Littorina littorea, and trace the evolutionary trajectories of the ancestral proto-complement repertoire.

Characterization of the gene encoding a fibrinogen-related protein expressed in Crassostrea gigas hemocytes.

Four exons of the CgFrep1 gene (3333 bp long) encode a putative fibrinogen-related protein (324 aa) bearing a single C-terminal FBG domain. Transcripts of the gene obtained from hemocytes of different Pacific oysters show prominent individual variation based on SNP and indels of tandem repeats resulted in polymorphism of N-terminus of the putative CgFrep1 polypeptide. The polypeptide chain bears N-terminal coiled-coil region potentially acting as inter-subunit interface in the protein oligomerization.

Membrane Attack Complex/Perforin domain-containing proteins in a dual-species transcriptome of caenogastropoda Littorina littorea and its trematode parasite Himasthla elongata.

The MACPF domain-containing proteins potentially able to build a transmembrane pore structure are found in the transcriptome of a common periwinkle kidney loaded with trematode rediae. Two homologs of mammalian Macrophage expressed gene 1 (Mpeg1), LlMpeg1-1 and LIMpeg1-2, share similar domain structure with the only difference such as LIMpeg1-2 transcript lacks of a C-terminal transmembrane helix. Expression of membrane-anchored protein LlMpeg1-1 is similar in kidneys of naturally infected with trematode Himasthla elongata and uninfected snails.

Lectin-like molecules in transcriptome of Littorina littorea hemocytes.

The common periwinkle Littorina littorea was introduced in the list of models for comparative immunobiology as a representative of phylogenetically important taxon Caenogastropoda. Using Illumina sequencing technology, we de novo assembled the transcriptome of Littorina littorea hemocytes from 182 million mRNA-Seq pair-end 100 bp reads into a total of 15,526 contigs clustered in 4472 unigenes. The transcriptome profile was analyzed for presence of carbohydrate-binding molecules in a variety of architectural contexts.

Himasthla elongata: Implantation of rediae to the specific iteroparous long-living host, Littorina littorea, results in the immune rejection.

All semelparous short-lived gastropods studied so far for the experimental transplantation of trematode parthenitae, from one specific host to another, showed more or less successful acceptance of implanted parasites. We implanted echinostomatid rediae, Himasthla elongata, to the specific iteroparous long-living host, coenogastropod Littorina littorea. Using simple and low-invasive implantation techniques we have tested 680 snails injected with 75 redia microhemipopulations (MHP) harvested from naturally infected snails.

Molecular cloning of α-2-macroglobulin from hemocytes of common periwinkle Littorina littorea.

We report the sequence of the proteinase inhibitor with a wide inhibition spectrum, α-2-macroglobulin (α2M), belonging to the thioester superfamily of proteins. This is the first α2M sequence from coenogastropod prosobranch snails. The full-length cDNA was cloned by RACE method, spans 7897 bp and contains an open reading frame of 5460 bp.

A new gene family of single fibrinogen domain lectins in Mytilus.

In molluscs haemolymph lectins bearing fibrinogen-like domain (FREP) act as immune pattern-recognition receptors. A full-length cDNAs of MytFREP1 and MytFREP2 cloned from haemocytes of blue mussel Mytilus edulis encoded putative polypeptides of 230 and 241 amino acids. Both polypeptides consist of signal peptide and C-terminal fibrinogen-like domain.

In search of the origin of FREPs: characterization of Aplysia californica fibrinogen-related proteins.

All haemolymph lectins with uniquely juxtaposed N-terminal domain similar to the immunoglobulin superfamily (IgSF) and C-terminal fibrinogen (FBG) termed FBG-related proteins (FREP) are documented till now only in the pulmonate mollusc Biomphalaria glabrata. Using genomic WGS database we have found two FREP genes from marine opistobranch Aplysia californica named AcFREP1 and AcFREP2. The AcFREP1 and AcFREP2 mRNA molecules have been subsequently isolated from cDNA of sea hare larvae as well as adult mollusc tissues.

Himasthla elongata: effect of infection on expression of the LUSTR-like receptor mRNA in common periwinkle haemocytes.

The first mollusc mRNA coding G-protein-coupled transmembrane receptor (GPcapital ES, CyrillicR), homologous to human receptors LUSTR 1 (GPR107) and LUSTR 2 (GPR108), was isolated from haemocytes of common periwinkle Littorina littorea. The analyses showed that the full-length cDNA is 1935 bp long and is predicted to encode a 614 amino acid protein (named Lit-LUSTR) with a calculated molecular mass of 69.6 kDa and theoretical isoelectric point 7.

The enigma of the haemogram "left-shift" in periwinkles infected with trematodes.

To make further progress in the understanding of digenean immune evasive tactics in their snail host, we compared the haemopoietic parameters and haemocyte functional potencies in the prosobranch Littorina littorea, which were either healthy or infected with echinostome trematode Himasthla elongata. The haemocyte concentration in the circulation of infected individuals was significantly higher than in uninfected ones, 3300 microl(-1) and 1882 microl(-1), respectively. Intense haemopoiesis in haemolymph of infected snails was evidenced by 4-fold higher BrdU incorporation into nuclei of haemocytes as well as elevated level of cyclin D expression in these cells.

[The presence of nitric oxide synthase in mucosal epithelial cells of the frog urinary bladder].

In experiments on frog Rana temporaria L. urinary bladder, we investigated localization of NO-synthase (NOS) in urinary bladder slices and measured NOS activity in the suspension of mucosal epithelial cells. Intensive NADPH-diaphorase staining which is widely used as an indicator of NOS activity was found in mucosal epithelium.

Modulation of mitogen-activated protein kinases (MAPK) activity in response to different immune stimuli in haemocytes of the common periwinkle Littorina littorea.

The modulation of mitogen-activated protein kinase (MAPK) activity in haemocytes of the common periwinkle (Littorina littorea) in response to immune challenges by lipopolysaccharide from Echerichia coli (LPS), mannan from baker's yeast Saccharomyces cerevisiae and secretory-excretory products (SEP) of trematodes Himasthla elongata (Echinostomatidae) or after the treatment with phorbol ester (PMA) has been studied by Western blotting using affinity purified rabbit polyclonal antibodies. Exposure of the cells in suspension to PMA, LPS and mannan triggered an activation of p38 and ERK2. The JNK-mediated cascade was modulated differently by the elicitors examined.

Rediae of echinostomatid and heterophyid trematodes suppress phagocytosis of haemocytes in Littorina littorea (Gastropoda: Prosobranchia).

A modulation of the phagocytic activity of hemocytes from the common periwinkle Littorina littorea by secretory-excretory products (SEP) released by trematode rediae during axenic in vitro cultivation was studied. The SEP released by the parasites Himasthla elongata (Echinostomatidae) and Cryptocotyle lingua (Heterophyidae) were found to inhibit the phagocytosis of zymozan particles by periwinkle hemocytes. The specificity of SEP effects was assessed: SEP of Himasthla militaris and Cryptocotyle concavum, two trematodes belonging to the same genera but infecting another closely related prosobranch snail Hydrobia ulvae, were also shown to be able to suppress L.