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Glens Falls NY 12801
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License #: 112058
Taxonomy Codes:207L00000X 207LP2900X
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The teleost acute-phase inflammatory response and caspase activation by a novel alarmin-like ligand. - Journal of leukocyte biology
This study tested the hypothesis that NCAMP-1 has alarmin-like properties and activates the caspase-1-binding site in cells of the teleost bone marrow (equivalent). In mammals, alarmins have been studied extensively; however, in teleosts, little is known about their identity and functions. Similar to alarmins, NCAMP-1 has a broad spectrum of bacteriolytic activity. NCAMP-1 is constitutively present in CF serum, and levels were increased following infection with Edwardsiella ictaluri. Binding to AK cells was determined with rNCAMP-1 and an anti-His-tag antibody. In vitro treatment of AK (bone marrow equivalent) or spleen cells with rNCAMP-1 increased the IL-1Î² message three- to fivefold at 3 h, 6 h, and 9 h post-treatment. The association of NCAMP-1 with the activities of alarmin ATP and the acute inflammatory response was demonstrated by NCAMP-1-induced P2X7R pore opening and YO-PRO-1 cellular influx. The association of NCAMP-1 binding with inflammasome activation was demonstrated by NCAMP-1 activation of the caspase-1-binding site for tetrapeptide Z-YVAD-FMK. In competition assays, this tetrapeptide competitively inhibited subsequent binding by the pan-caspase substrate tripeptide FAM-VAD-FMK. Lymphocyte-like cells from the spleen were 16%(+), and epithelial cells were also positive for NCAMP-1. IHC staining and confocal microscopy confirmed the cytosolic existence of NCAMP-1 in lymphoreticular tissue and IL-1Î² in AK cells. CF T cell lines G14D and 28S.3 expressed NCAMP-1 in the cytosol and in storage granules. These studies strongly suggested that NCAMP-1 is an alarmin-like ligand with similar but distinct activities to those of ATP and HMGB-1.
Foaling rates after surgical repair of ventral cervical lacerations using a Trendelenburg position in 18 anesthetized mares. - Veterinary surgery : VS
To (1) describe a surgical technique for repair of lacerations of the ventral aspect of the cervix in anesthetized mares in Trendelenburg position; and (2) report conception rates and foaling after repair.Case series.Mares (nâ€‰=â€‰18).Cervical lacerations were repaired in 2 layers with anesthetized mares in Trendelenburg position. The cervix was retracted caudally with stay-sutures, and after excision of the healed margin of the laceration, the internal cervical mucosa and fibromuscular tissue were sutured in 1 layer with a continuous Lembert pattern. The external cervical mucosa was sutured using a simple-continuous suture pattern. Number of foals produced by each mare before and after surgical repair was obtained from client/farm communication and from a commercial record keeping service.Mean repair time for 1 laceration was 22 minutes and for 2 lacerations was 29 minutes. Nine mares conceived and 7 had at least 1 live healthy foal.Trendelenburg position allows for efficient repair of a ventrally located cervical laceration in anesthetized mares.Â© Copyright 2013 by The American College of Veterinary Surgeons.
Humoral response of captive zebra sharks Stegostoma fasciatum to salivary gland proteins of the leech Branchellion torpedinis. - Fish & shellfish immunology
Parasitism by the marine leech Branchellion torpedinis is known to cause disease and mortality in captive elasmobranchs and is difficult to control when inadvertently introduced into public aquaria. Preliminary characterization of the salivary gland transcriptome of B.Â torpedinis has identified anticoagulants, proteases, and immunomodulators that may be secreted into host tissues to aid leech feeding. This retrospective study examined antigen-specific serum IgM responses in captive zebra sharks Stegostoma fasciatum to leech salivary gland extract. Antibody response was examined by ELISA and Western blot assays in 20 serum samples from six zebra sharks, with a 5 year history of leech infection, and 18 serum samples from 8 captive bred zebra sharks, with no history of leech exposure. ELISA demonstrated significantly higher serum IgM titers to salivary gland extract in exposed zebra sharks compared to the non-exposed population. No obvious trends in antibody titers were appreciated in exposed zebra sharks over a four-year period. One-dimensional and two-dimensional Western blot assays revealed IgM targeted specific salivary gland proteins within the 40, 55, 70 and 90Â kD range. Antigenic proteins identified by liquid chromatography-tandem mass spectrometry and de novo peptide sequencing include a secreted disintegrin, metalloproteinase and thrombospondin motif containing protein (ADAMTS), tubulin, aldehyde dehydrogenase and two unknown proteins. Humoral immune responses to leech salivary gland proteins warrants further investigation as there may be options to exploit immune mechanisms to reduce parasite burdens in aquaria.Copyright Â© 2012 Elsevier Ltd. All rights reserved.
Methamphetamine use, dependence and treatment access in rural and regional North Coast of New South Wales, Australia. - Drug and alcohol review
To identify the sociodemographic, health, drug use patterns, treatment coverage and barriers to treatment among regular methamphetamine users in rural and regional North Coast of New South Wales.A structured questionnaire was used to measure sociodemographic factors, health and well-being, drug use patterns, methamphetamine dependence, engagement in methamphetamine treatment and barriers to treatment. Participants were 140 regular methamphetamine users. Dependent and non-dependent participants were compared to identify factors associated with dependence.Participants were predominantly in their thirties, male and had low levels of education, high levels of unemployment and polydrug use. Participants who were dependent on methamphetamine (59%) were more likely to report impaired mental health and to have been diagnosed with depression, anxiety and drug-induced psychosis. One quarter of dependent methamphetamine users had received treatment in the last year and half had ever received treatment. The main barriers to receiving treatment were a lack of perceived need or motivation to seek treatment and concerns about confidentiality.Methamphetamine users living on the North Coast of New South Wales require treatment options tailored to address a complex array of physical and psychological problems. The findings highlight the need for psychiatric support and improved coordination between mental health and drug and alcohol services in rural and regional areas.
Identification of phagocytic cells, NK-like cytotoxic cell activity and the production of cellular exudates in the coelomic cavity of adult zebrafish. - Developmental and comparative immunology
Coelomic cavity (CC) cells of mature zebrafish harvested by lavage with media or trypsin-EDTA contained 0.80-1.20 x 10(5) and 2.0-3.5 x 10(5) cells, respectively. Media lavage was composed of granulocytes (60-80%), lymphocytes (10-20%), and NCC (4-10%). Granulocytes had large electron dense cytoplasmic paracrystalline granules and a segmented nucleus; they expressed plastin-1, myeloid specific peroxidase and MCSF mRNA; and they were NCAMP-1(+). Lymphocytes had B- and T-cell specific mRNA and were NCAMP-1(-) and NCCRP-1(-). NCC were 3 microm, NCAMP-1(+) and NCCRP-1(+) and did not express B- and T-cell specific mRNA. Additionally, trypsin lavage contained monocytes (marginated chromatin, low nuclear:cytoplasm ratio, sparse cytosolic granules) and macrophages (non-segmented nuclei, no margination of chromatin, abundant electron dense granules). E. coli injected into the CC were phagocytosed in a dose and time dependent fashion by granulocytes, monocytes and macrophages. NCC lysed mammalian target cells and NCAMP-1 expressing hybridoma cells in redirected lysis assays.
Cellular expression and antimicrobial function of a phylogenetically conserved novel histone 1x-like protein on mouse cells: a potential new class of pattern recognition receptor. - Journal of leukocyte biology
A H1x-like protein (i.e., NCAMP-1) is expressed on the membrane and in GEs from fish NK-like cells. In the present study, we identify the imprinting control region mouse NCAMP-1 ortholog using NCAMP-1 polyclonal antibodies and mAb. Polychromatic flow cytometry revealed NCAMP-1 expression on PBLs (Gr-1(+) PMNs were 21.1% NCAMP-1(+); DX-5(+) NK cells were 12.2% NCAMP-1(+)), mesenteric LN cells (CD11c(+) DCs were 23.2% NCAMP-1(+); Gr-1(+) PMNs were 24.8% NCAMP-1(+); CD21(+) B cells were 17.8% NCAMP-1(+)), and splenocytes (CD11c(+) were 39.6% NCAMP-1(+); Gr-1(+) PMNs were 40.9% NCAMP-1(+); DX-5(+) NK cells were 24.3% NCAMP-1(+); CD21(+) B cells were 28.5% NCAMP-1(+)). Western blot analysis using pNCAMP-1 and GEs from RAW 264.7 cells produced a 32-kDa signal. GEs from RAW 264.7 cells produced a significant reduction in Escherichia coli CFU. This antimicrobial killing activity was inhibited by pretreatment of the extract with (polyclonal) anti-NCAMP-1. Treatment with preimmune serum did not reduce bacterial cell killing. Confocal microscopy using NCAMP-1 and LAMP-1 mAb demonstrated that NCAMP-1 was located on the membrane and in cytosolic vesicles of RAW 264.7 cells and did not appear to colocalize with LAMP-1. NCAMP-1 may participate as a bifunctional protein on cells. It is expressed on the membranes of phagocytic cells, NK cells, and APCs in mice as well as in the granules of macrophages. In phagocytic cells, NCAMP-1 may participate in a nonregulated exocytosis pathway of cellular secretion.
Role of nonspecific cytotoxic cells in bacterial resistance: expression of a novel pattern recognition receptor with antimicrobial activity. - Molecular immunology
Pattern recognition receptors (PRR) recognize invariant bacterial, viral, protozoan and certain synthetic ligands. PRR may be expressed as outer membrane (or endosomal) or cytosolic proteins and function to signal cell activation processes during inflammation responses. In the present study, a novel membrane receptor, NCC cationic antimicrobial protein-1 (NCAMP-1), is described that is expressed on nonspecific cytotoxic cell (NCC) membranes and is found in granule extracts from these cells. In recombinant form, full-length (amino acids 1-203) and truncated N (NT; amino acids 1-60) and C (CT; amino acids 116-203) terminal forms of NCAMP-1 had antibacterial activity against bovine, avian and lab strain Escherichia coli. Recombinant NCAMP-1-NT also killed the gram-negative fish pathogen Edwardsiella ictaluri. Maximal bacterial killing of a representative avian E. coli, APEC 3721, occurred at 60min post-treatment with 2microg/ml of rNCAMP-1-NT. Killing occurred by NCAMP-1-NT-induced alterations in the permeability of the bacterial cell wall. Polyclonal antibody anti-NCAMP-1 specifically neutralized the antimicrobial activity of recombinant NCAMP-1-NT against E. coli APEC 3751. Expression of NCAMP-1 as a NCC membrane protein was analyzed by flow cytometry using anti-NCAMP-1 monoclonal antibody 9C9. Merged images from immunofluorescence microscopy showed that NCAMP-1 and the NCC receptor protein (NCCRP-1) are co-expressed on NCC membranes. NCAMP-1 was identified in acetic acid granule extracts of NCC by Western blot analysis using polyclonal anti-NCAMP-1 and killing of E. coli by these extracts was specifically inhibited by this polyclonal. These data suggested that NCAMP-1 is a membrane protein and may participate in antibacterial innate immunity by granule exocytosis during inflammatory responses in teleosts.
The effects of obesity and fatty acids on the feline immune system. - Veterinary immunology and immunopathology
Obesity is a rising problem in cats. It is a risk factor for several diseases and has been linked to impaired immunity. The goal of this study was to determine the effect of body composition and effects of diet on immune function in cats. Twenty-eight short-term obese and 12 lean cats with equal gender distribution were evenly and randomly divided into two groups which were either fed a diet containing saturated (SFA) or long-chain n-3 polyunsaturated fatty acids (3-PUFA) for a period of 6 months prior to testing. Blood was collected by venipuncture from the jugular vein. Blood samples were analyzed in a double blind fashion. A complete blood count was performed and lymphocyte distribution was examined by flow cytometric analysis with specific fluorescein-conjugated subset markers. Immune function was measured as follows: the proliferative activity of different cellular fractions was tested with polyclonal mitogens such as lipopolysaccharide (LPS), phytohaemagglutinin (PHA), phorbol 12-myristate 13-acetate (PMA), Ca ionophore, and concanavalin A. Innate immune functions assessed were phagocytosis and natural killer cell (NK) cytotoxicity. A similar immune innate and adaptive immune response was elicited regardless of diet or body condition. However, there was no correlation between body condition, diet, and any of the quantitative and qualitative functional responses of the immune system. We conclude that short-term obesity and the fatty acid composition of the diet do not alter immune responses in cats.
Molecular characterization and expression of a granzyme of an ectothermic vertebrate with chymase-like activity expressed in the cytotoxic cells of Nile tilapia (Oreochromis niloticus). - Immunogenetics
We have identified the gene coding for a novel serine protease with close similarities to mammalian granzymes from nonspecific cytotoxic cells of a teleost fish Oreochromis niloticus. The genomic organization of tilapia granzyme-1 (TLGR-1) has the signature five-exon-four-intron structure shared by all granzymes and similar hematopoietic Ser proteases. Molecular modeling studies suggested a granzyme-like structure for this protein with four disulfide linkages and two additional Cys residues. The expression of this gene is found to be restricted to cytotoxic cell populations with a low level of constitutive expression when compared to similar granzymes in other teleost species. High levels of transcriptional activation of TLGR-1 with different stimuli suggested that this gene is highly induced during immune reactions. Triplet residues around the active site Ser of TLGR, which determines the primary substrate specificity of granzymes, differ significantly from that of other granzymes. Recombinant TLGR-1 was expressed in the mature and proenzyme forms using pPICZ-alpha vector in the Pichia pastoris expression system. Recombinant TLGR-1 was used to determine the primary substrate specificity of this protease using various synthetic thiobenzyl ester substrates. In vitro enzyme kinetics assays suggested a preference for residues with bulky side chains at the P1 site, indicating a chymase-like activity for this protease. These results indicate the presence of novel granzymes in cytotoxic cells from ectothermic vertebrates.
Molecular cloning of cellular apoptosis susceptibility (CAS) gene in Oreochromis niloticus and its proposed role in regulation of non-specific cytotoxic cell (NCC) functions. - Fish & shellfish immunology
Cellular apoptosis susceptibility (CAS) gene is a homologue of the chromosome segregation gene (CSE) in yeast, involved in multiple cellular mechanisms associated with cell proliferation as well as cell death. CAS is highly expressed in proliferating cells but at a lower level in quiescent cells and tissues. Therefore it appears that CAS may play an important role in cancer development. We have previously identified CAS in tilapia non-specific cytotoxic cells (NCC) with a cross-reacting monoclonal antibody. Its expression was up-regulated in NCC in response to apoptosis regulatory factors. In the present report, the molecular cloning and expression of CAS in NCC is described, suggesting the importance of this protein in regulation of teleost immune functions. Furthermore, CAS expression is proposed as one of the mechanisms of regulation of activation induced programmed cell death (AIPCD) in these cytotoxic cells. As CAS expression is ubiquitous, we expect that these studies will help identify proliferating cells protected from apoptosis in additional tissues.
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