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The avian heterophil.  


Heterophils play an indispensable role in the immune defense of the avian host. To accomplish this defense, heterophils use sophisticated mechanisms to both detect and destroy pathogenic microbes. Detection of pathogens through the toll-like receptors (TLR), FC and complement receptors, and other pathogen recognition receptors has been recently described for the avian heterophil. Upon detection of pathogens, the avian heterophil, through a network of intracellular signaling pathways and the release and response to cytokines and chemokines, responds using a repertoire of microbial killing mechanisms including production of an oxidative burst, cellular degranulation, and production of extracellular matrices of DNA and histones (HETs). In this review, the authors describe the recent advances in our understanding of the avian heterophil, its functions, receptors and signaling, identified antimicrobial products, cytokine and chemokine production, and some of the effects of genetic selection on heterophils and their functional characteristics. PMID:23583524

Genovese, Kenneth J; He, Haiqi; Swaggerty, Christina L; Kogut, Michael H



Avian Heterophils in Inflammation and Disease Resistance  

Microsoft Academic Search

Heterophils are the predominate granu- lated leukocyte in the acute inflammatory response in gallinaceous birds. Heterophils are highly phagocytic and are capable of a broad spectrum of antimicrobial activity. They accumulate in inflamed tissue, causing tissue damage and forming heterophil granulomas that are morphologically similar to inflammatory lesions in reptiles. The avian heterophil lacks myeloperoxidase and depends primarily on nonoxidative



Heterophilic antibody interference in immunometric assays.  


Immunometric assays are inherently vulnerable to interference from heterophilic antibodies, endogenous antibodies that bind assay antibodies. The consequences of such interference can be devastating. In this review, we discuss strategies that reduce the damage caused by heterophilic antibodies. Clinicians should only order blood tests that are indicated for the patient and clinical setting at hand, and have the confidence to question laboratory results discordant with the clinical picture. Laboratorians should familiarize themselves with the vulnerability of the assays they offer, and be able to perform and interpret adequate confirmatory measures correctly. When designing immunoassays, the immunoassay industry should invest the necessary resources in specific protective measures against heterophilic antibody interference. Examples include using antibody fragments and the addition of effective blockers to assay reagents. The increasing use of modified monoclonal mouse antibodies both in therapy and diagnostics could present a particular challenge in the future. PMID:24094636

Bolstad, Nils; Warren, David J; Nustad, Kjell



Similarities between heterophilic and homophilic cadherin adhesion  

PubMed Central

The mechanism that drives the segregation of cells into tissue-specific subpopulations during development is largely attributed to differences in intercellular adhesion. This process requires the cadherin family of calcium-dependent glycoproteins. A widely held view is that protein-level discrimination between different cadherins on cell surfaces drives this sorting process. Despite this postulated molecular selectivity, adhesion selectivity has not been quantitatively verified at the protein level. In this work, molecular force measurements and bead aggregation assays tested whether differences in cadherin bond strengths could account for cell sorting in vivo and in vitro. Studies were conducted with chicken N-cadherin, canine E-cadherin, and Xenopus C-cadherin. Both qualitative bead aggregation and quantitative force measurements show that the cadherins cross-react. Furthermore, heterophilic adhesion is not substantially weaker than homophilic adhesion, and the measured differences in adhesion do not correlate with cell sorting behavior. These results suggest that the basis for cell segregation during morphogenesis does not map exclusively to protein-level differences in cadherin adhesion.

Prakasam, A. K.; Maruthamuthu, V.; Leckband, D. E.



Reducing heterophilic antibody interference in immunoassays using single chain antibodies  

SciTech Connect

Sandwich ELISA microarrays have the potential to simultaneously quantify the levels of multiple diagnostic targets in a biological sample. However, as seen with traditional ELISA diagnostics, heterophilic antibodies (HA) in patient sera have the potential to cause interference in these assays. We demonstrate here that reducing the diagnostic capture antibody to its minimal functional unit, the variable heavy and light domains artificially connected with a short polypeptide linker (scFv), is an effective strategy for reducing the HA assay interference.

Baird, Cheryl L.; Tan, Ruimin; Fischer, Christopher J.; Victry, Kristin D.; Zangar, Richard C.; Rodland, Karin D.



Differential cytokine mRNA expression in heterophils isolated from Salmonella-resistant and -susceptible chickens  

PubMed Central

We recently showed that increased in vitro heterophil functional efficiency translates to increased in vivo resistance to a systemic Salmonella enteritidis (SE) infection utilizing a parental pair of broiler chickens (lines A and B) and the F1 reciprocal crosses (C and D). Heterophils produce cytokines and modulate acute protection against Salmonella in young poultry. Therefore, we hypothesize that heterophils from SE-resistant chickens (A and D) have the ability to produce an up-regulated pro-inflammatory cytokine response compared to that of heterophils from SE-susceptible chickens (B and C). In this study, heterophils were isolated from day-old chickens and treated with either RPMI-1640 (as the control), or phagocytic agonists (SE, or SE opsonized with either normal chicken serum or immune serum against SE) and cytokine mRNA expression assessed using real-time quantitative reverse transcription–polymerase chain reaction. Heterophils from SE-resistant chickens (A and D) had significantly higher levels of pro-inflammatory cytokine (interleukin (IL)-6, IL-8, and IL-18) mRNA expression upon treatment with all agonists compared to heterophils from SE-susceptible lines (B and C). Further, heterophils from SE-resistant chickens had significantly decreased mRNA expression levels of transforming growth factor-?4, an anti-inflammatory cytokine, when compared to heterophils from SE-susceptible chickens. These data indicate cytokine gene expression in heterophils may be a useful parameter in determining resistance to Salmonella, as indicated by our previous in vivo SE studies. Therefore, heterophil functional efficiency and cytokine production may be useful biomarkers for poultry breeders to consider when developing new immunocompetent lines of birds.

Swaggerty, Christina L; Kogut, Michael H; Ferro, Pamela J; Rothwell, Lisa; Pevzner, Igal Y; Kaiser, Pete



Heterophile antibodies indicate progression of autoimmunity in human type 1 diabetes mellitus before clinical onset.  


We previously reported serum cytokines in a group of long term non-progressors to Type 1 diabetes; this reactivity detected in ELISA is now identified as heterophile antibody in some sera. Here, we characterize heterophile antibody activity. A 14 kDa-polypeptide from heterophile antibody containing serum bound to an anti-IL-4 column, but IL-4 was not detected by Western blot or by MS/MS sequencing. However, in 2/13 heterophile antibody positive sera, T-cell growth was potentiated and was blocked by an anti-human immunoglobulin. To examine the relationship between low affinity heterophile antibody presence and disease progression, 1100 archived serum samples were analyzed with two pairs of antibodies from 443 diabetes-free first degree relatives of Type 1 diabetes mellitus patients for heterophile antibody; 95 individuals developed diabetes on follow-up. Twenty-two individuals, whose serum was heterophile antibody positive with the second pair of antibodies (but negative with the first pair of antibodies), had a significantly higher incidence of developing diabetes after five years. Thirty-seven individuals with heterophile antibody reactivity with the first pair of antibodies, regardless of reactivity with the second pair of antibodies, had a significantly lower incidence of developing diabetes. While we cannot exclude the presence of genuine cytokine in all sera, these data indicate the presence of distinct groups of heterophile antibodies in patients at high risk to develop diabetes. Thus, anti-Ig heterophilic antibodies with different immunochemical reactivities are linked to the progression of or protection from Type 1 diabetes autoimmunity. PMID:11905851

Orban, T; Kent, S C; Malik, P; Milner, J D; Schuster, K; Jackson, R A; Hafler, D A



Heterophil functional responses to dietary immunomodulators vary in genetically distinct chicken lines.  


The effect of dietary supplementation of immunomodulators on in vitro chicken heterophil function was investigated using three diverse genetic lines of chickens (broiler, Fayoumi, and Leghorn). Dietary supplementation with ?-glucan, ascorbic acid, and corticosterone was fed from 8 to 11 weeks of age. Heterophil function was evaluated weekly during supplementation using phagocytosis, bacterial killing, and heterophil extracellular traps (HETs)-DNA release. Fayoumis fed the basal diet had significantly higher HETs-DNA release (P=0.002) than Leghorns and broilers. Both genetic line and immunomodulator diet supplementation had significant effects on bacterial killing (line and diet effect: P<0.001) and HETs-DNA release (line: P<0.001; diet: P=0.043). Dietary supplementation with immunomodulators, therefore, shows potential to affect and augment heterophil function in chickens. The current results also suggest the important role of genetics in innate immune responses. PMID:21645931

Chuammitri, Phongsakorn; Redmond, Sarah B; Kimura, Kayoko; Andreasen, Claire B; Lamont, Susan J; Pali?, Dušan



Comparison of MAP and tyrosine kinase signaling in heterophils from commercial and wild-type turkeys.  


Protein tyrosine and mitogen-activated kinases are crucial mediators of the host innate immune response, conferring signals from surface receptors on the host cell to the nucleus of the cell where gene expression occurs. Heterophils were isolated from wild-type Rio Grande turkeys and a commercial line of turkeys (Line A) on days 4 and 7 post-hatch. Heterophils were stimulated for 1h with Salmonella enteritidis (SE) or opsonized SE (OPSE). After stimulation, cells were lysed and lysates were tested for mitogen-activated protein kinase (MAPK) activity. Specifically, phosphorylation of extracellular signal-regulated protein kinase (ERK1/2) and p38 MAPK phosphorylation were assayed using commercially available ELISA kits. Total protein tyrosine kinase (PTK) activity was also assayed. On both days 4 and 7 post-hatch, heterophils from Rio Grande turkeys had significantly higher levels of ERK 1/2 and p38 MAPK kinase activity upon stimulation with either SE or OPSE (p<0.001). Likewise, PTK values on days 4 and 7 in Rio Grande turkey heterophils were significantly higher upon stimulation with SE than with OPSE and were significantly (p<0.001) higher than PTK levels in Line A upon SE and OPSE stimulation. The data presented supports previous heterophil functional comparison studies wherein heterophils from Rio Grande turkeys had higher levels of oxidative burst and degranulation activities as compared to the activity observed in commercial Line A heterophils. This suggests that the regulation and control of these functions are mediated by protein tyrosine and mitogen-activated kinases. Furthermore, the data suggest that selection of commercial lines of turkeys for larger, heavier bodies, and faster growth may be associated with subsequent selection for decreased innate immune functions related to intracellular signaling mechanisms and possibly a subsequent increase in susceptibility to disease. PMID:17241661

Genovese, Kenneth J; He, Haiqi; Lowry, Virginia K; Kogut, Michael H



Functional annotation of proteomic data from chicken heterophils and macrophages induced by carbon nanotube exposure.  


With the expanding applications of carbon nanotubes (CNT) in biomedicine and agriculture, questions about the toxicity and biocompatibility of CNT in humans and domestic animals are becoming matters of serious concern. This study used proteomic methods to profile gene expression in chicken macrophages and heterophils in response to CNT exposure. Two-dimensional gel electrophoresis identified 12 proteins in macrophages and 15 in heterophils, with differential expression patterns in response to CNT co-incubation (0, 1, 10, and 100 µg/mL of CNT for 6 h) (p < 0.05). Gene ontology analysis showed that most of the differentially expressed proteins are associated with protein interactions, cellular metabolic processes, and cell mobility, suggesting activation of innate immune functions. Western blot analysis with heat shock protein 70, high mobility group protein, and peptidylprolyl isomerase A confirmed the alterations of the profiled proteins. The functional annotations were further confirmed by effective cell migration, promoted interleukin-1? secretion, and more cell death in both macrophages and heterophils exposed to CNT (p < 0.05). In conclusion, results of this study suggest that CNT exposure affects protein expression, leading to activation of macrophages and heterophils, resulting in altered cytoskeleton remodeling, cell migration, and cytokine production, and thereby mediates tissue immune responses. PMID:24823882

Li, Yun-Ze; Cheng, Chung-Shi; Chen, Chao-Jung; Li, Zi-Lin; Lin, Yao-Tung; Chen, Shuen-Ei; Huang, San-Yuan



Functional Annotation of Proteomic Data from Chicken Heterophils and Macrophages Induced by Carbon Nanotube Exposure  

PubMed Central

With the expanding applications of carbon nanotubes (CNT) in biomedicine and agriculture, questions about the toxicity and biocompatibility of CNT in humans and domestic animals are becoming matters of serious concern. This study used proteomic methods to profile gene expression in chicken macrophages and heterophils in response to CNT exposure. Two-dimensional gel electrophoresis identified 12 proteins in macrophages and 15 in heterophils, with differential expression patterns in response to CNT co-incubation (0, 1, 10, and 100 ?g/mL of CNT for 6 h) (p < 0.05). Gene ontology analysis showed that most of the differentially expressed proteins are associated with protein interactions, cellular metabolic processes, and cell mobility, suggesting activation of innate immune functions. Western blot analysis with heat shock protein 70, high mobility group protein, and peptidylprolyl isomerase A confirmed the alterations of the profiled proteins. The functional annotations were further confirmed by effective cell migration, promoted interleukin-1? secretion, and more cell death in both macrophages and heterophils exposed to CNT (p < 0.05). In conclusion, results of this study suggest that CNT exposure affects protein expression, leading to activation of macrophages and heterophils, resulting in altered cytoskeleton remodeling, cell migration, and cytokine production, and thereby mediates tissue immune responses.

Li, Yun-Ze; Cheng, Chung-Shi; Chen, Chao-Jung; Li, Zi-Lin; Lin, Yao-Tung; Chen, Shuen-Ei; Huang, San-Yuan



Horse erythrocyte glycoprotein-latex reagent that reacts with infectious mononucleosis heterophile antibody.  

PubMed Central

A sialoglycoprotein from horse erythrocytes was isolated in essentially homogeneous form and found to contain the neuraminidase-sensitive determinant of the horse erythrocyte for Paul-Bunnell heterophile antibodies of infectious mononucleosis. This reactivity was retained after covalent coupling of the antigen to latex particles. The latex reagent has greater stability (greater than 3 years) than either fresh or preserved horse erythrocytes. It can be used in a direct slide test; no absorption of the serum is necessary. The new test compared favorably with some standard tests for infectious mononucleosis antibody which use horse erythrocytes. Agreement of the latex test with the absorbed fresh horse cell test was 100%, and agreement with a stabilized horse erythrocyte spot test was 94%. The latex test also agreed 100% with a radioimmunoassay for the detection of heterophile antibody with 125I-labeled horse erythrocyte glycoprotein. The new latex test was compared with a bovine erythrocyte glycoprotein-latex test, and a correlation of 94% was observed. In addition, it was compared with the Wöllner test (enzyme-treated sheep cell-absorbed sheep cell agglutination test) with which it agreed in 92% of samples tested.

Fletcher, M A; Caldwell, K E; Saez, L; Latif, Z



Homophilic and heterophilic binding activities of Nr-CAM, a nervous system cell adhesion molecule  

PubMed Central

Nr-CAM is a membrane glycoprotein that is expressed on neurons. It is structurally related to members of the N-CAM superfamily of neural cell adhesion molecules having six immunoglobulin-like domains and five fibronectin type III repeats in the extracellular region. We have found that the aggregation of chick brain cells was inhibited by anti-Nr-CAM Fab' fragments, indicating that Nr-CAM can act as a cell adhesion molecule. To clarify the mode of action of Nr-CAM, a mouse fibroblast cell line L-M(TK-) (or L cells) was transfected with a DNA expression construct encoding an entire chicken Nr-CAM cDNA sequence. After transfection, L cells expressed Nr-CAM on their surface and aggregated. Aggregation was specifically inhibited by anti-Nr-CAM Fab' fragments. To check the specificity of this aggregation, a fusion protein (FGTNr) consisting of glutathione S-transferase linked to the six immunoglobulin domains and the first fibronectin type III repeat of Nr- CAM was expressed in Escherichia coli. Addition of FGTNr to the transfected cells blocked their aggregation. Further analysis using a combination of cell aggregation assays, binding of cells to FGTNr- coated substrates, aggregation of FGTNr-coated Covaspheres and binding of FGTNr-coated Covaspheres to FGTNr-coated substrates revealed that Nr- CAM mediates two types of cell interactions: a homophilic, divalent cation-independent binding, and a heterophilic, divalent cation- dependent binding. Homophilic binding was demonstrated between transfected L cells, between chick embryo brain cells and FGTNr, and between Covaspheres to which FGTNr was covalently attached. Heterophilic binding was shown to occur between transfected and untransfected L cells, and between FGTNr and primary chick embryo fibroblasts; in all cases, it was dependent on the presence of either calcium or magnesium. Primary chick embryo glia or a human glial cell line did not bind to FGTNr-coated substrates. The results indicate that Nr-CAM is a cell adhesion molecule of the nervous system that can bind by two distinct mechanisms, a homophilic mechanism that can mediate interactions between neurons and a heterophilic mechanism that can mediate binding between neurons and other cells such as fibroblasts.



Reversible adrenal insufficiency and heterophile antibodies in a case of autoimmune polyendocrinopathy syndrome  

PubMed Central

A 27-year-old male was admitted with diabetic ketoacidosis and altered sensorium with slurring of speech and ataxia. He was managed with intravenous insulin and fluids and later shifted to basal bolus insulin regimen and during further evaluation was diagnosed to be suffering from primary hypothyroidism and adrenal insufficiency. He was started on thyroxin replacement and steroids only during stress. After three months of follow up he was clinically euthyroid. His glycemic control was adequate on oral anti-hyperglycemic drugs and adrenal insufficiency recovered. However, his thyrotropin levels were persistently elevated on adequate replacement doses of thyroxin. His repeat TSH was estimated after precipitating serum with polyethylene glycol which revealed normal TSH. Here we report reversible adrenal insufficiency with hypothyroidism with falsely raised TSH because of presence of heterophile antibodies in a case of poly glandular endocrinopathy syndrome.

Kharb, Sandeep; Gundgurthi, Abhay; Dutta, Manoj K.; Garg, M. K.



Severe histiolymphocytic and heterophilic bronchopneumonia as a reaction to in ovo fowlpox vaccination in broiler chicks.  


Broiler chickens on several farms from a single poultry company experienced neurological signs and mortality in chicks between 3 days and 10 days of age over a 3-week period after use of a fowlpox-vectored infectious laryngotracheitis virus vaccine in ovo. At necropsy the lungs contained numerous tan or gray, opaque to translucent, 0.5- to 2.0-mm nodules in the parenchyma. Microscopic lesions were a multifocal severe lymphohistiocytic and heterophilic bronchopneumonia. Immunohistochemistry was positive for fowlpox virus in macrophages and lymphocytes, and polymerase chain reaction on paraffin-embedded lung tissues was positive for a fowlpox vector virus commonly used as a vaccine. The cause of the neurological signs was not determined. PMID:20080499

Williams, S M; Smith, J A; Garcia, M; Brinson, D; Kiupel, M; Hofacre, C



Reversible adrenal insufficiency and heterophile antibodies in a case of autoimmune polyendocrinopathy syndrome.  


A 27-year-old male was admitted with diabetic ketoacidosis and altered sensorium with slurring of speech and ataxia. He was managed with intravenous insulin and fluids and later shifted to basal bolus insulin regimen and during further evaluation was diagnosed to be suffering from primary hypothyroidism and adrenal insufficiency. He was started on thyroxin replacement and steroids only during stress. After three months of follow up he was clinically euthyroid. His glycemic control was adequate on oral anti-hyperglycemic drugs and adrenal insufficiency recovered. However, his thyrotropin levels were persistently elevated on adequate replacement doses of thyroxin. His repeat TSH was estimated after precipitating serum with polyethylene glycol which revealed normal TSH. Here we report reversible adrenal insufficiency with hypothyroidism with falsely raised TSH because of presence of heterophile antibodies in a case of poly glandular endocrinopathy syndrome. PMID:24910843

Kharb, Sandeep; Gundgurthi, Abhay; Dutta, Manoj K; Garg, M K



Heterophil to Lymphocyte Ratio and Tonic Immobility Reactions to Preslaughter Handling in Broiler Chickens Treated with Ascorbic Acid  

Microsoft Academic Search

Stress and fear responses were evaluated in broiler chicks that were pretreated for 24 h with 0 ppm (control) or 1,200 ppm of L-ascorbic acid (AA) in their drinking water. The birds were subsequently subjected to either upright handling (UH) or inverted (IH) handling for about 45 s. Heterophil (H) counts, lymphocyte (L) counts, and H\\/L ratios (H\\/L) ratios were

I. Zulkifli; M. T. Che Norma; C. H. Chong; T. C. Loh


Human heterophile antibodies recognizing distinct carbohydrate epitopes on basidiolipids from different mushrooms.  


Investigating the immune properties of basidiolipids, i.e., glycoinositolphosphoceramides (GIPC) of basidiomytes, higher mushrooms, it was detected that sera of normal adult human subjects contained IgG2 and IgM heterophile antibodies (hetAbs) that immunoreacted with these lipids. However, this immune recognition was not shared by the glycolipids of all mushroom species. The basidiolipids of Amanita virosa (eng., death cup) and Cantharellus cibarius (engl., chantarelle), of all mushroom species studied, did not bind antibodies of normal human sera. In addition, only certain basidiolipids of the other mushroom species that have been investigated, i.e., Agaricus bisporus (engl., field mushroom), Calvatia exipuliformis engl., puffball), Lentinus edodes (jap., Shiitake), Leccinum scabrum (engl., red birch boletus), and Pleurotus ostreatus (engl., oyster mushroom), immunoreacted with the human hetAbs. The basidiolipids that were recognized by the human hetAbs had either terminal Galalpha1-6Gal < or Galbeta1-6Man< epitopes. Enzymatic destruction of the respective carbohydrate epitopes abolished the previous immune reactivity. It is assumed that contact with non human antigens causes generation of the anti-basidiolipid antibodies. PMID:11465669

Jennemann, R; Sandhoff, R; Gröne, H J; Wiegandt, H



Protein tyrosine kinase and mitogen-activated protein kinase signalling pathways contribute to differences in heterophil-mediated innate immune responsiveness between two lines of broilers.  


Protein tyrosine phosphorylation mediates signal transduction of cellular processes with protein tyrosine kinases (PTKs) regulating virtually all signalling events. The mitogen-activated protein kinase (MAPK) super-family consists of three conserved pathways that convert receptor activation into cellular functions: extracellular response kinases (ERK), c-Jun N-terminal kinases (JNK) and p38. Previously conducted studies using two chicken lines (A and B) show line A heterophils are functionally more responsive and produce a differential cytokine/chemokine profile compared with line B, which also translates to increased resistance to bacterial challenges. Therefore, we hypothesize the differences between the lines result from distinctive signalling cascades that mediate heterophil function. Heterophils from lines A and B were isolated from 1-day-old chickens and total phosphorylated PTK and p38, JNK, ERK, and transcription factor (activator protein 1 (AP-1) and nuclear factor kappa B (NF-?B)) protein levels quantified following interaction with Salmonella Enteritidis (SE). Control and SE-treated heterophils from line A had greater (P?0.05) PTK phosphorylation compared to line B with increased (P?0.05) activation of p38. Conversely, line B heterophils activated JNK (P?0.05). There were no differences in ERK between control and activated heterophils for either line. Defined signalling inhibitors were used to show specificity. The AP-1 and NF-?B transcription factor families were also examined, and c-Jun and p50, respectively, were the only members different between the lines and both were up-regulated in line A compared with line B. These data indicate that increased responsiveness of line A heterophils is mediated, largely, by an increased ability to activate PTKs, the p38 MAPK pathway and specific transcription factors, all of which directly affect the innate immune response. PMID:21711188

Swaggerty, C L; He, H; Genovese, K J; Pevzner, I Y; Kogut, M H



Laying hen behavior 2. Cage type preference and heterophil to lymphocyte ratios.  


Studies were conducted to determine hen preference for and stress response to cage type. By using a plywood (1.25 cm) test apparatus with open- and solid-sided compartments and a plexiglass divider at the entrance for controlling passage, birds (n = 20) were evaluated as to their choice of compartment after training and acclimation. For each test, after training, an individual bird was placed in the start box and given 1 min to acclimate before making a choice. The experiment was repeated after rotating the apparatus 180 degrees. In Experiment 1, the open-sided compartment was chosen by 45% of the hens, and the solid-sided compartment by 25% (P = 0.02); 30% chose neither. In Experiment 2, the compartment with open sides was chosen by 70% of the hens, and that with solid sides was chosen by 15% (P = 0.004); 15% chose neither. The length of time required to choose one compartment over the other did not differ in either experiment (P = 0.29; P = 0.76). In Experiment 3, tests were videotaped from 0830 h to 1330 h, and tapes were scored for time spent in each compartment. Birds were observed to spend more time in open- compared with solid-sided compartments (P = 0.02). To assess stress level of birds exposed to each type of enclosure, blood was collected from 24 commercial Hy-Line W36 hens housed long-term in either open (n = 12) or solid (n = 12) cages, and heterophil:lymphocyte (H:L) ratios were determined. Birds in solid cages had higher H:L ratios than did birds in open cages (P = 0.02), indicating a greater stress response. These data would suggest that hens may prefer greater visual access to their surroundings afforded by open cages. PMID:10780641

Elston, J J; Beck, M; Alodan, M A; Vega-Murillo, V



Gene Expression Analysis of Toll-Like Receptor Pathways in Heterophils from Genetic Chicken Lines that Differ in Their Susceptibility to Salmonella enteritidis.  


Previously conducted studies using two chicken lines (A and B) show that line A birds have increased resistance to a number of bacterial and protozoan challenges and that heterophils isolated from line A birds are functionally more responsive. Furthermore, when stimulated with Toll-like receptor (TLR) agonists, heterophils from line A expressed a totally different cytokine and chemokine mRNA expression pattern than heterophils from line B. A large-scale gene expression profile using an Agilent 44K microarray on heterophils isolated from line A and line B also revealed significantly differential expression in many immune-related genes following Salmonella enteritidis (SE) stimulation, which included genes involved in the TLR pathway. Therefore, we hypothesize the differences between the lines result from distinctive TLR pathway signaling cascades that mediate heterophil function and, thus, innate immune responsiveness to SE. Using quantitative RT-PCR on mRNA from heterophils isolated from control and SE-stimulated heterophils of each line, we profiled the expression of all chicken homologous genes identified in a reference TLR pathway. Several differentially expressed genes found were involved in the TLR-induced My88-dependent pathway, showing higher gene expression in line A than line B heterophils following SE stimulation. These genes included the TLR genes TLR4, TLR15, TLR21, MD-2, the adaptor proteins Toll-interleukin 1 receptor domain-containing adaptor protein (TIRAP), Tumor necrosis factor-receptor associated factor 3 (TRAF3), the I?B kinases transforming growth factor-?-activating kinase 1 (TAK1), IKK? and IKK?, the transcription factors NFkB2 and interferon regulatory factor 7, phosphatidylinositol-3 kinase (PI-3K), and the mitogen-activated protein kinase p38. These results indicate that higher expression of TLR signaling activation of both MyD88-dependent and TRIF-dependent pathways are more beneficial to avian heterophil-mediated innate immunity and a complicated regulation of downstream adaptors is involved in stronger induction of a TLR-mediated innate response in the resistant line A. These findings identify new targets for genetic selection of chickens to increase resistance to bacterial infections. PMID:22783275

Kogut, Michael H; Chiang, Hsin-I; Swaggerty, Christina L; Pevzner, Igal Y; Zhou, Huaijun



Gene Expression Analysis of Toll-Like Receptor Pathways in Heterophils from Genetic Chicken Lines that Differ in Their Susceptibility to Salmonella enteritidis  

PubMed Central

Previously conducted studies using two chicken lines (A and B) show that line A birds have increased resistance to a number of bacterial and protozoan challenges and that heterophils isolated from line A birds are functionally more responsive. Furthermore, when stimulated with Toll-like receptor (TLR) agonists, heterophils from line A expressed a totally different cytokine and chemokine mRNA expression pattern than heterophils from line B. A large-scale gene expression profile using an Agilent 44K microarray on heterophils isolated from line A and line B also revealed significantly differential expression in many immune-related genes following Salmonella enteritidis (SE) stimulation, which included genes involved in the TLR pathway. Therefore, we hypothesize the differences between the lines result from distinctive TLR pathway signaling cascades that mediate heterophil function and, thus, innate immune responsiveness to SE. Using quantitative RT-PCR on mRNA from heterophils isolated from control and SE-stimulated heterophils of each line, we profiled the expression of all chicken homologous genes identified in a reference TLR pathway. Several differentially expressed genes found were involved in the TLR-induced My88-dependent pathway, showing higher gene expression in line A than line B heterophils following SE stimulation. These genes included the TLR genes TLR4, TLR15, TLR21, MD-2, the adaptor proteins Toll-interleukin 1 receptor domain-containing adaptor protein (TIRAP), Tumor necrosis factor-receptor associated factor 3 (TRAF3), the I?B kinases transforming growth factor-?-activating kinase 1 (TAK1), IKK? and IKK?, the transcription factors NFkB2 and interferon regulatory factor 7, phosphatidylinositol-3 kinase (PI-3K), and the mitogen-activated protein kinase p38. These results indicate that higher expression of TLR signaling activation of both MyD88-dependent and TRIF-dependent pathways are more beneficial to avian heterophil-mediated innate immunity and a complicated regulation of downstream adaptors is involved in stronger induction of a TLR-mediated innate response in the resistant line A. These findings identify new targets for genetic selection of chickens to increase resistance to bacterial infections.

Kogut, Michael H.; Chiang, Hsin-I; Swaggerty, Christina L.; Pevzner, Igal Y.; Zhou, Huaijun



Effect of heat and several additives related to stress levels on fluctuating asymmetry, heterophil:lymphocyte ratio, and tonic immobility duration in White Leghorn chicks.  


The purpose of this study was to analyze the effects of heat and several additives related to stress on fluctuating asymmetry (groups 1 to 10), heterophil:lymphocyte ratio (groups 1 to 3 and 8 to 10), and tonic immobility duration (groups 1 to 7 and 10) in White Leghorn chicks at 42 d of age. Chicks in group 1 (heat) were reared with temperatures 8°C greater than those of the control group. Groups 2 to 9 consisted of chicks reared with temperatures 8°C greater than those of the control group and addition of capsaicin, allicin, ascorbic acid, tryptophan, brewer's yeast, lactic acid, corticosterone, or cholesterol in diet. Chicks in group 10 (control) were reared with standard temperatures. Heat effect was significant (P<0.05) for the heterophil:lymphocyte ratio, which was greater in heat-stressed chicks without any additives and smaller in control chicks. There were no significant differences for the fluctuating asymmetry and the tonic immobility duration between both groups. Heterophil:lymphocyte ratio for heat-stressed chicks with capsaicin or allicin was significantly lower (P<0.05) than that of heat-stressed chicks without any additives. Capsaicin effect was not significant for the fluctuating asymmetry and the tonic immobility duration, whereas allicin significantly increased fluctuating asymmetry of wing length and tonic immobility duration (P<0.05). The addition of lactic acid or corticosterone resulted in greater fluctuating asymmetry of wing length of heat-stressed chicks (P<0.05). In conclusion, an increased heterophil:lymphocyte ratio was found in heat-stressed chicks without additives, indicating that it is a more reliable indicator of the effect of heat in chicks. In addition, dietary capsaicin or allicin supplementation was effective to alleviate the stress induced by the high temperature, as indicated by a lower heterophil:lymphocyte ratio. PMID:20852096

Prieto, M T; Campo, J L



Mixed infection with Libyostrongylus dentatus and Libyostrongylus douglassii induces a heterophilic inflammatory infiltrate in the proventriculus of ostriches.  


Libyostrongylus dentatus and Libyostrongylus douglassii are haematophagous nematodes found in the proventriculus and the ventriculus of ostriches. Pathological damage leading to bird death has been attributed to L. douglassii. However, histopathology of the mixed infection has not been reported. The aim of the present work was to characterize the cellular inflammatory infiltrate found in the proventriculus of ostriches with a mixed infection. Analysis of the collected nematodes confirmed a mixed infection in the proventriculus of examined birds. Histopathological examination of the proventriculus showed an inflammatory infiltrate composed of granular cells in close proximity to the nematodes. The granulocyte infiltrate was composed mainly of heterophils identified by the lack of peroxidase and presence of fusiform granules. PMID:21812715

de Andrade, Josiana Gomes; de Carvalho, Eulógio Carlos Queiroz; Santos, Clóvis de Paula; DaMatta, Renato Augusto



Heterophilic Binding of L1 on Unmyelinated Sensory Axons Mediates Schwann Cell Adhesion and Is Required for Axonal Survival  

PubMed Central

This study investigated the function of the adhesion molecule L1 in unmyelinated fibers of the peripheral nervous system (PNS) by analysis of L1- deficient mice. We demonstrate that L1 is present on axons and Schwann cells of sensory unmyelinated fibers, but only on Schwann cells of sympathetic unmyelinated fibers. In L1-deficient sensory nerves, Schwann cells formed but failed to retain normal axonal ensheathment. L1-deficient mice had reduced sensory function and loss of unmyelinated axons, while sympathetic unmyelinated axons appeared normal. In nerve transplant studies, loss of axonal-L1, but not Schwann cell-L1, reproduced the L1-deficient phenotype. These data establish that heterophilic axonal-L1 interactions mediate adhesion between unmyelinated sensory axons and Schwann cells, stabilize the polarization of Schwann cell surface membranes, and mediate a trophic effect that assures axonal survival.

Haney, C.A.; Sahenk, Z.; Li, C.; Lemmon, V.P.; Roder, J.; Trapp, B.D.



Automodification of arginine-specific ADP-ribosyltransferase purified from chicken peripheral heterophils and alteration of the transferase activity.  


Auto-ADP-ribosylation of arginine-specific ADP-ribosyltransferase purified from chicken peripheral heterophils was investigated. When the purified ADP-ribosyltransferase was analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by Coomassie brilliant blue staining, two protein bands corresponding to the molecular masses of 27.5 and 28.0 kDa were detected. Both proteins were auto-ADP-ribosylated when they were examined by zymographic in situ gel assay without exogenous acceptor for ADP-ribose transfer. The automodification was inhibited by the acceptor, arginine or agmatine, and an inhibitor of arginine-specific ADP-ribosyltransferase, novobiocin. The ADP-ribose-transferase linkage was labile in 0.5 M hydroxylamine (pH 7.5). The automodified transferase was not chased by a large excess of nonradioactive NAD and did not catalyze transfer of its ADP-ribose to p33, an endogenous substrate protein for the transferase in heterophils, therefore, that automodified transferase cannot serve as an intermediate in ADP-ribosylation of other proteins. Auto-ADP-ribosylated transferase showed higher activity than did the unmodified transferase in catalyzing ADP-ribosylation of the basic acceptor such as poly(L-arginine) and p33 while to ADP-ribosylate the acidic proteins such as casein, the modified transferase was less active. Automodification of the transferase decreased polyanion-induced ADP-ribosylation of p33. Automodification of arginine-specific ADP-ribosyltransferase apparently alters the specificity of its own substrate. PMID:8311468

Yamada, K; Tsuchiya, M; Nishikori, Y; Shimoyama, M



Spatial and Temporal Variation in Heterophil-to-Lymphocyte Ratios of Nestling Passerine Birds: Comparison of Blue Tits and Great Tits  

PubMed Central

Environmental factors affecting trophic conditions act as stressors on nestling altricial birds. Access of parental birds to a sufficient supply of food in a limited period of the nestling stage differ in time and space, depending on nesting habitat, prey density and weather conditions. Heterophil-to-lymphocyte ratio (H/L) is considered as a reliable indicator of prolonged stress reaction in birds. In this study we examine if variation in H/L shows consistent spatio-temporal patterns in nestlings of two parids, blue tit Cyanistes caeruleus and great tit Parus major. We found that blue tit nestlings had on average higher H/L than great tit nestlings, which corresponds with the ecological sensitivity of these species. In both species H/L was higher in a poor parkland habitat than in a high quality forest habitat. In nestling blue tits, higher H/L values occurred in years characterized by more extreme weather conditions and worse caterpillar availability. Such consistent patterns of variation in the H/L ratio of nestling blue tits and great tits suggest that, when age-dependent effects are controlled, the ratio can be used as an indicator of physiological stress that is generated by food-related stressors differing in space and time. In particular, elevated H/L ratios are indicative of human-induced changes in the structure of breeding habitats.

Banbura, Jerzy; Skwarska, Joanna; Banbura, Miroslawa; Gladalski, Michal; Holysz, Magdalena; Kalinski, Adam; Markowski, Marcin; Wawrzyniak, Jaroslaw; Zielinski, Piotr



Neurofascin induces neurites by heterophilic interactions with axonal NrCAM while NrCAM requires F11 on the axonal surface to extend neurites  

PubMed Central

Neurofascin and NrCAM are two axon-associated transmembrane glycoproteins belonging to the L1 subgroup of the Ig superfamily. In this study, we have analyzed the interaction of both proteins using neurite outgrowth and binding assays. A neurofascin-Fc chimera was found to stimulate the outgrowth of tectal cells when immobilized on an inert surface but not as a soluble form using polylysine as substrate. Antibody blocking experiments demonstrate that neurite extension on immobilized neurofascin is mediated by NrCAM on the axonal surface. Under the reverse experimental conditions where NrCAM induces neurite extension, F11, and not neurofascin, serves as axonal receptor. Binding studies using transfected COS7 cells and immunoprecipitations reveal a direct interaction between neurofascin and NrCAM. This binding activity was mapped to the Ig domains within neurofascin. The neurofascin-NrCAM binding can be modulated by alternative splicing of specific stretches within neurofascin. These studies indicate that heterophilic interactions between Ig-like proteins implicated in axonal extension underlie a regulation by the neuron.



The effects of long-term caging and molt of Single Comb White Leghorn hens on heterophil to lymphocyte ratios, corticosterone and thyroid hormones.  


Two commercial strains of 18-wk-old Single Comb White Leghorn (SCWL) hens, HyLine W-36 and DeKalb XL, were housed six hens per cage in layer cages at two densities (361 and 482 cm2 per bird) with two replications each per strain and density combination. The high density treatment contained 24 hens per replication, and the low density treatment contained 18 hens per replication. Egg production was measured during the first egg production cycle, a molt (fast) period, and the first 4 wk of the second lay cycle (20 to 68 wk of age). Blood samples were obtained from six hens from each replicate in each strain and density combination (total of 48) at 20, 26, 34, 43, 51, 62, 64, and 68 wk of age. In addition, blood samples were obtained in a random order from hens in each cage, and the sequence of sampling was recorded (1 to 6). Blood smears were made, from which heterophil to lymphocyte ratios (H:L) were determined. Radioimmunoassays were conducted to determine levels of plasma corticosterone (CS), 3,5,3'-triiodothyronine (T3), and thyroxine (T4). The results indicated that strain did not affect percentage hen-day egg production (%HDP). Strain and cage density did not affect H:L, T3, T4, or CS. However, these parameters were affected by bird age, which was related to the egg production cycle. Plasma CS significantly (P < or = 0.001) increased during peak %HDP at 26 wk and 64 wk during the molt (fast), and H:L significantly (P < or = 0.001) increased during the molt (fast) at 64 wk. The sequence in which blood samples were obtained, from hens within a cage in sampling order, also increased plasma CS. The CS was significantly (P < or = 0.001) elevated in the third, fifth, and sixth hens from which blood samples were drawn. Plasma T3 and T4 changed during the production cycle. The T3 was significantly (P < or = 0.0001) depressed during peak egg production at 26 wk and during the molt (fast) at 64 wk when compared with the other time periods. Plasma T4 was depressed (P < or = 0.0001) at 51 wk and was elevated (P < or = 0.0001) at 64 wk during the molt (fast). The physiological and metabolic parameters of the different hen strains and cage densities were similar during egg production. However, CS, T3, T4, and H:L changed with age in relation to the egg production cycle. In addition, the physiological demands of peak egg production and molt (fast) appeared to be similar. PMID:10780646

Davis, G S; Anderson, K E; Carroll, A S



Effects of dietary yeast extract on turkey stress response and heterophil oxidative burst activity  

Microsoft Academic Search

1. Effective nutritional approaches to counteract the negative effects of stress may provide food animal producers with useful alternatives to antibiotics. In this study, turkeys were fed on a standard diet, or the same diet supplemented with yeast extract (YE), to determine if YE would improve disease resistance in a stress model.2. At 16 weeks of age, half of the

G. R. Huff; V. Dutta; W. E. Huff; N. C. Rath



Direct Ca2+-dependent Heterophilic Interaction between Desmosomal Cadherins, Desmoglein and Desmocollin, Contributes to Cell-Cell Adhesion  

PubMed Central

Human fibrosarcoma cells, HT-1080, feature extensive adherens junctions, lack mature desmosomes, and express a single known desmosomal protein, Desmoglein 2 (Dsg2). Transfection of these cells with bovine Desmocollin 1a (Dsc1a) caused dramatic changes in the subcellular distribution of endogenous Dsg2. Both cadherins clustered in the areas of the adherens junctions, whereas only a minor portion of Dsg2 was seen in these areas in the parental cells. Deletion mapping showed that intact extracellular cadherin-like repeats of Dsc1a (Arg1-Thr170) are required for the translocation of Dsg2. Deletion of the intracellular C-domain that mediates the interaction of Dsc1a with plakoglobin, or the CSI region that is involved in the binding to desmoplakin, had no effect. Coimmunoprecipitation experiments of cell lysates stably expressing Dsc1a with anti-Dsc or -Dsg antibodies demonstrate that the desmosomal cadherins, Dsg2 and Dsc1a, are involved in a direct Ca2+-dependent interaction. This conclusion was further supported by the results of solid phase binding experiments. These showed that the Dsc1a fragment containing cadherin-like repeats 1 and 2 binds directly to the extracellular portion of Dsg in a Ca2+-dependent manner. The contribution of the Dsg/ Dsc interaction to cell–cell adhesion was tested by coculturing HT-1080 cells expressing Dsc1a with HT-1080 cells lacking Dsc but expressing myc-tagged plakoglobin (MPg). In the latter cells, MPg and the endogenous Dsg form stable complexes. The observed specific coimmunoprecipitation of MPg by anti-Dsc antibodies in coculture indicates that an intercellular interaction between Dsc1 and Dsg is involved in cell–cell adhesion.

Chitaev, Nikolai A.; Troyanovsky, Sergey M.



Human antibodies recognizing the envelope glycoprotein of the baboon endogenous virus BaEV are of heterophil origin  

Microsoft Academic Search

Human sera were previously shown to possess antibodies capable of recognizing purified retrovirus envelope glycoproteins. In an extension of earlier studies we investigated sera from various groups of patients for an immune reaction against purified glycoprotein of the baboon endogenous virus BaEV. Reproducible demonstrations of oncovirus-like particles in human teratocarcinomas focused our main interest on sera from patients with testicular

Dorothee Wernicke; Reinhard Kurth



Homophilic and heterophilic polycystin 1 interactions regulate E-cadherin recruitment and junction assembly in MDCK cells  

PubMed Central

Summary Autosomal dominant polycystic kidney disease (ADPKD) is the most common inherited human renal disease and is caused by mutations in two genes, PKD1 (85%) and PKD2 (15%). Cyst epithelial cells are characterised by a complex cellular phenotype including changes in proliferation, apoptosis, basement membrane composition and apicobasal polarity. Since polycystin 1 (PC1), the PKD1 protein, has been located in the basolateral membrane of kidney epithelial cells, we hypothesised that it might have a key role in mediating or stabilising cell-cell interactions. In non-ciliated L929 cells, stable or transient surface expression of the PC1 extracellular domain was sufficient to confer an adhesive phenotype and stimulate junction formation. In MDCK cells, we found that PC1 was recruited to the lateral membranes coincident with E-cadherin within 30 minutes after a `calcium switch'. Recruitment of both proteins was significantly delayed when cells were treated with a PC1 blocking antibody raised to the PKD domains. Finally, PC1 and E-cadherin could be coimmunoprecipitated together from MDCK cells. We conclude that PC1 has a key role in initiating junction formation via initial homophilic interactions and facilitates junction assembly and the establishment of apicobasal polarity by E-cadherin recruitment.

Streets, Andrew J.; Wagner, Bart E.; Harris, Peter C.; Ward, Christopher J.; Ong, Albert C. M.



GENETICS Influence of Perches and Footpad Dermatitis on Tonic Immobility and Heterophil to Lymphocyte Ratio of Chickens  

Microsoft Academic Search

The purpose of the present study was to analyze the effect of perches and footpad dermatitis on stress and fear levels of chickens. In experiment 1 hetero- phil to lymphocyte ratio and tonic immobility duration were measured in 36-wk-old hens (36 birds) from 2 Span- ish breeds of chickens (Black Menorca and Quail Castellana) housed in pens with or without

J. L. Campo; M. G. Gil; S. G. Davila; I. Munoz


IMMUNOLOGY, HEALTH, AND DISEASE Stress Response Differences and Disease Susceptibility Reflected by Heterophil to Lymphocyte Ratio in Turkeys Selected for Increased Body Weight1  

Microsoft Academic Search

Three genetic lines of turkeys were com- pared for their responses to Escherichia coli challenge fol- lowing dexamethasone injection (Dex) or E. coli challenge preceding transport stress (TS). The turkey lines were a slow growing line selected for increased egg production (Egg line), a fast growing line selected for increased 16- wk BW (F line), and a commercial line (Comm

G. R. Huff; W. E. Huff; J. M. Balog; N. C. Rath; N. B. Anthony; K. E. Nestor


Tissue Culture Studies of Human Lymphocyte in Infectious Mononucleosis.  

National Technical Information Service (NTIS)

The site of production for the heterophile antibody in patients with infectious mononucleosis is unknown. Atypical lymphocytes from patients with infectious mononucleosis were grown in tissue culture in an attempt to demonstrate heterophile antibody produ...

R. A. Gams C. A. Coltman



Mononucleosis spot test  


Monospot test; Heterophile antibody test; Heterophile agglutination test; Paul-Bunnell test; Forssman antibody test ... The mononucleosis spot test is done when symptoms of mononucleosis are ... Fatigue Fever Large spleen (possibly) Sore throat Tender ...


The Effect of Different Feed Restriction Programs and Dietary L-Carnitine Supplementation on Hepatic Lipogenesis, Plasma Heterophil to Lymphocyte Ratio and Yolk IgY Content of Broiler Breeder Hens  

Microsoft Academic Search

Two experiments were conducted to determine effects of Everyday (ED) or Skip-a-day (SK) feeding and dietary L-carnitine on lipid metabolism and stress in broiler breeders. In Experiment 1 a 2x2 factorial design was used to compare feeding regimens (ED vs SK) and L-carnitine supplementation (0 vs 50 mg\\/kg). L-carnitine supplementation began at d 1 and lasted throughout the 45 weeks

M. de Beer; C. N. Coon



Hematological parameters in brood-rearing great tits in relation to habitat, multiple breeding and sex  

Microsoft Academic Search

Very little is known about the causes and correlates for variation of individual condition in the wild. However, such knowledge is essential for understanding the mechanisms that mediate environmental effects to populations. We studied the variation of several hematological condition indices (hematocrit, albumin, globulin and triglyceride concentrations, albumin\\/globulin ratio, lymphocyte and heterophile concentrations and heterophile\\/lymphocyte ratio) and body mass in

Priit Kilgas; Raivo Mänd; Marko Mägi; Vallo Tilgar



Effects of post-hatching feeding management (fasting) on blood parameters of broilers  

Microsoft Academic Search

hematocrit volume (HCT, %) , mean corpuscular volume (MCV, fL), hemoglobin concentration (HGB, g\\/dL), and specific lymphocytes count (L, %) and heterophil (H, %) to determine heterophil\\/lymphocyte ratio (H\\/L). PGC = 191.22 - 1.91271 T + 0.03504 T2 + 0.62285 A (R2 =0.2675); PPC (females) = 1.22 + 0.00480 T + 0.02558 A - 0.00015 A2 (R2 =0.2488); PPC (males)



Diagnostic Evaluation of Mononucleosis-Like Illnesses  

Microsoft Academic Search

Clinicians face a diagnostic challenge when a patient with the classic fever, pharyngitis, and lymphadenopathy triad of infectious mononucleosis has a negative “spot” heterophile antibody test. This screening test, although commonly considered sensitive for the presence of Epstein-Barr virus (EBV) infection, may be negative early after infection. A growing number of pathogens have been reported to cause heterophile-negative mononucleosis-like illnesses,

Christopher Hurt; Dominick Tammaro



Expression and purification of an Epstein-Barr virus encoded 23-kDa protein and characterization of its immunological properties  

Microsoft Academic Search

Serodiagnosis of Epstein-Barr virus (EBV) infection is currently based on the detection of antibodies to distinct EBV antigens by immunofluorescence and enzyme-linked immunosorbent assay-based tests, or in part on the detection of heterophile antibodies by the Paul-Bunnell-Davidson heterophile assay. In the past few years, the specificity and the sensitivity of serodiagnostic assay systems has been improved considerably by the use

Udo Reischl; Christian Gerdes; Manfred Motz; Hans Wolf



Erythrophagocytosis in circulating blood of loggerhead turtles Caretta caretta: the pitting of Heinz bodies.  


In humans red blood cells live on average of 120 days. Senescent erythrocytes are removed from circulation by macrophages; this process, called erythrophagocytosis (EPC), takes place mainly in the liver but also in the bone marrow. We studied this process in loggerhead turtles Caretta caretta, inhabiting the Mediterranean Sea. Leukocytes of reptiles include heterophils, eosinophils, lymphocytes, basophils, thrombocytes, monocytes, and of these heterophils and monocytes represent the main phagocytic cells. The aim of this study was to investigate the blood of C. caretta specimens and to observe, in the blood smears, heterophils phagocytizing erythrocytes. The phagocytizing cells were also identified by cytochemical reactions. Furthermore some examples of tear-shaped erythrocytes (dacrocytes) were observed. A mechanism presuming a causative link between erythrophagocytosis and teardrop-erythrocyte detection have been proposed to explain this seemingly general pattern of all specimens both wild and housed. PMID:24254579

Ruggiero, Maria Grazia; Ferretti, Luigi; Glomski, Chester; Pica, Alessandra



Feed intake alters immune cell functions and ovarian infiltration in broiler hens: implications for reproductive performance.  


Leukocytes are known to participate in ovarian activities in several species, but there is a surprising lack of information for the common chicken. Broiler hens consuming feed ad libitum (AL) exhibit a number of ovarian irregularities, but leukocyte functions are unstudied. In contrast to feed-restricted (R) hens, AL feeding for 7 wk significantly reduced egg production and clutch length while increasing pause length and atretic follicle numbers (P < 0.05). Granulosa cells from F1 follicles of AL hens contained less progesterone, and follicle walls were thicker with loose fibrous morphology and had less collagenase-3-like gelatinolytic activity but more IL-1beta (P < 0.05) production, suggestive of slower maturation in ovulatory process and inflamed necrosis. Interestingly, while highly infiltrated with immune cells, particularly heterophils, IL-1beta, MMP-22-like, and gelatinase A activities were reduced in AL hen peripheral heterophils and monocytes (P < 0.05); however, AL monocytes showed an increase in phagocytosis rate (P < 0.05). Generation of reactive oxygen intermediates was also suppressed in AL heterophils but increased in AL monocytes (P < 0.05). In contrast to leukocyte-free control, both AL and R heterophils and monocytes suppressed progesterone production and increased cell death in a dose-dependent manner when coincubated with granulosa cells at different ratios (P < 0.05). AL monocytes suppressed progesterone production more, but AL heterophils were less proapoptotic when compared to their R counterparts (P < 0.05). Alterations of cellular ceramide content (P < 0.05) corresponded to the discrepancy between heterophil and monocyte functionality. In conclusion, leukocyte dysfunction contributes to impaired ovarian activities of overfed broiler hens. PMID:24829031

Liu, Zu-Chen; Xie, Yi-Lun; Chang, Chai-Ju; Su, Chia-Ming; Chen, Yu-Hui; Huang, San-Yuan; Walzem, Rosemary L; Chen, Shuen-Ei



Effects of glycerol on performance, egg traits, some blood parameters and antibody production to SRBC of laying hens  

Microsoft Academic Search

This study was designed to investigate the effects of the usage of glycerol from biodiesel production from soybean oil in laying hen diets on laying performance, egg traits, heterophils to lymphocytes ratio (H\\/L), some blood parameters and antibody production to SRBC. A total of 180 Lohmann Brown laying hens 39weeks of age were allocated to four dietary treatments with one

Sakine Yalç?n; Handan Erol; Bülent Özsoy; ?lyas Onba??lar; Suzan Yalç?n; Aykut Üner



Characterisation of the green turtle's leukocyte subpopulations by flow cytometry and evaluation of their phagocytic activity.  


Phagocytosis is a fundamental aspect of innate immunity that is conserved across many species making it a potentially useful health-assessment tool for wildlife. In non-mammalian vertebrates, heterophils, monocytes, macrophages, melanomacrophages, and thrombocytes all have phagocytic properties. Recently, B lymphocytes from fish, amphibians, and aquatic turtles have also showed phagocytic capacity. Phagocytes can be studied by flow cytometry; however, the use of this tool is complicated in reptiles partly because nucleated erythrocytes complicate the procedure. We separated green turtle leukocytes by density gradient centrifugation and identified subpopulations by flow cytometry and confocal microscopy. Additionally, we assessed their ability to phagocytize Fluorspheres and Ovoalbumin-Alexa. We found that heterophils and lymphocytes but not monocytes could be easily identified by flow cytometry. While heterophils from adults and juvenile turtles were equally able to phagocytize fluorspheres, adults had significantly more phagocytic ability for OVA-Alexa. Lymphocytes had a mild phagocytic activity with fluorospheres (27-38 %; 39-45 %) and OVA-Alexa (35-46 %; 14-22 %) in juvenile and adult green turtles, respectively. Confocal microscopy confirmed phagocytosis of fluorospheres in both heterophils and lymphocytes. This provides the first evidence that green turtle lymphocytes have phagocytic activity and that this assay could potentially be useful to measure one aspect of innate immunity in this species. PMID:24570347

Muñoz, F A; Franco-Noguez, S Y; Gonzalez-Ballesteros, E; Negrete-Philippe, A C; Flores-Romo, L



Tree swallow reproductive investment, stress, and parasites  

Microsoft Academic Search

We reduced or increased tree swallow, Tachycineta bicolor (Vieillot, 1808), clutch sizes by three eggs (50% of modal clutch size) to test experimentally for relationships between stress and parasite loads. In the first year of the study (1996), we enumerated two measures of stress (granulocyte to non-granulocyte ratios and heterophil to lympho- cyte ratios), blood parasites, and ectoparasites living on

Dave Shutler; Adele Mullie; Robert G. Clark



Nonspecific cellular defense of the avian respiratory system: a review  

Microsoft Academic Search

The normal, steady-state, avian respiratory system has very low numbers of residing avian respiratory phagocytes (ARP). Birds must rely heavily on the influx of ARP to defend against infectious agents. The system is refractory to elicitation by inert stimulants, but responds efficiently to replicating bacteria, with very rapid influx of large numbers of activated ARP (polymorphonuclear neutrophils, heterophils, and macrophages)

Thomas E. Toth



Salpingitis in geese associated with Mycoplasma sp. strain 1220  

Microsoft Academic Search

An outbreak of disease in a White Rhine laying goose flock was characterized by increased water uptake, increased mortality, production of eggs with abnormal shells, a 25% drop in egg production and 40% embryo mortality. Affected dead or sacrificed birds had sero-fibrinogranulocytic peritonitis and salpingitis, infiltration of the lamina propria in the uterus and heterophil granulocytes in the isthmus and

Mihály Dobos-Kovács; Zsuzsanna Varga; György Czifra; László Stipkovits



Cutaneous myelolipoma in a peach-faced lovebird (Agapornis roseicollis).  


A case of cutaneous myelolipoma in a 2.5-year-old female peach-faced lovebird is reported. Histopathologically, the subcutaneous mass on the tip of the left wing consisted of well circumscribed adipose tissue containing several aggregates of * haematopoietic cells. Heterophilic differentiation was most prominent in them. PMID:18645843

Ozakt, K; Kinoshita, H; Kurasho, H; Narama, I



Biochemical and hematological reference ranges for Amazon freshwater turtle, Podocnemis expansa (Reptilia: Pelomedusidae), with morphologic assessment of blood cells.  


Blood parameters are useful to measure physiological disturbances in chelonians. Thus they can provide important information for the diagnosis and prognosis of diseases. The aim of this paper was to determine the reference range of plasma glucose, total protein, triglycerides, cholesterol and urea levels, as well as hematocrit, hemoglobin, red blood cell, thrombocyte and white blood cell counts for healthy Podocnemis expansa (Schweigger, 1812), bred in a turtle farm in the Amazonas State, Brazil. Plasma glucose, hemoglobin, mean corpuscular volume (MCV), lymphocytes and heterophils were the parameters with the smallest variations. Significant (p<0.001) correlations between red blood cells count and hematocrit and hemoglobin concentration were found. Turtle P. expansa had lymphocytes, azurophils, heterophils, eosinophils and basophils with morphologic features similar to the ones of others turtles species. No investigated blood parameter was influenced by sex. In further studies, the established reference ranges might be useful for the health assessment of this turtle species. PMID:18617203

Oliveira-Júnior, A A; Tavares-Dias, M; Marcon, J L



Effects of an S6 strain of Mycoplasma gallisepticum inoculation before beginning of lay on the leukocytic characteristics of commercial layers.  


A clinical study was conducted on commercial layers housed in biological isolation units, within which exogenous stress factors potentially affecting bird performance were minimized. This set-up was devised in order to assess how a pre-lay inoculation of S6 strain Mycoplasma gallisepticum affects the leukocytic properties of laying chickens. Previous studies have demonstrated relative decreases in lymphocyte and relative increases in heterophil percentages in birds infected with other strains of Mycoplasma gallisepticum. However, current results showed that the differential percentages of lymphocytes were decreased, whereas those of heterophils were increased, in both sham-inoculated control birds and birds inoculated with S6 Mycoplasma gallisepticum between 19 and 26 wk of age. This study clearly shows that a pre-lay inoculation of S6 Mycoplasma gallisepticum alone had no apparent effect on the leukocyte profile of commercial layers housed in biological isolation units. PMID:15077815

Peebles, E D; Parker, T A; Branton, S L; Willeford, K O; Jones, M S; Gerard, P D; Pharr, G T; Maslin, W R



Cadherin-dependent mechanotransduction depends on ligand identity but not affinity  

PubMed Central

Summary This study investigates the relationship between classical cadherin binding affinities and mechanotransduction through cadherin-mediated adhesions. The mechanical properties of cadherin-dependent intercellular junctions are generally attributed to differences in the binding affinities of classical cadherin subtypes that contribute to cohesive energies between cells. However, cell mechanics and mechanotransduction may also regulate intercellular contacts. We used micropipette measurements to quantify the two-dimensional affinities of cadherins at the cell surface, and two complementary mechanical measurements to assess ligand-dependent mechanotransduction through cadherin adhesions. At the cell surface, the classical cadherins investigated in this study form both homophilic and heterophilic bonds with two-dimensional affinities that differ by less than threefold. In contrast, mechanotransduction through cadherin adhesions is strongly ligand dependent such that homophilic, but not heterophilic ligation mediates mechanotransduction, independent of the cadherin binding affinity. These findings suggest that ligand-selective mechanotransduction may supersede differences in cadherin binding affinities in regulating intercellular contacts.

Tabdili, Hamid; Langer, Matthew; Shi, Quanming; Poh, Yeh-Chuin; Wang, Ning; Leckband, Deborah



Fluorescence of the natural dye saffron: Selective reaction with eosinophil leucocyte granules  

Microsoft Academic Search

Treatment of methanol-fixed chicken, rat, horse and human blood smears with saturated solutions of saffron in borate buffer at pH 10 results in a bright yellow-green fluorescence reaction of the acidophilic cytoplasm granules in mammalian eosinophils and chicken heterophils under violet-blue exciting light. Spectral characteristics of saffron (emission peak at 543 nm under 436 nm excitation) and its selective fluorescence

Clara Isabel Trigoso; Juan Carlos Stockert



Clinical pathology results from cranes with experimental West Nile Virus infection  

USGS Publications Warehouse

Sandhill cranes (Grus canadensis) were vaccinated for and then challenged with West Nile virus. Resulting titers demonstrated protection in the vaccinated-challenged cranes as compared to the unvaccinated-challenged cranes. Clinical pathology results showed challenged cranes, whether vaccinated or not, had a decrease in their hematocrits and an elevation of 2.5-fold in their white blood cell counts as compared to unchallenged control sandhill cranes. No differences were apparent in the differential counts of heterophils and lymphocytes.

Olsen, Glenn H.



Inactivation of the N-CAM gene in mice results in size reduction of the olfactory bulb and deficits in spatial learning  

Microsoft Academic Search

NEURAL-CELL adhesion molecules (N-CAMs) are members of the immunoglobulin superfamily mediating homo- and heterophilic cell-cell interactions. N-CAM exists in various isoforms which are generated by alternative splicing1-3. During embryonic development, N-CAMs are expressed in derivatives of all three germ layers, whereas in the adult animal they are predominantly present in neural tissue. Processes like neurulation4, axonal outgrowth5, histogenesis of the

Harold Cremer; Rita Lange; Annette Christoph; Markus Plomann; Gaby Vopper; Jürgen Roes; Russell Brown; Stanley Baldwin; Philipp Kraemer; Stephen Scheff; Dagmar Barthels; Klaus Rajewsky; Wolfgang Wille



Plasmin-sensitive Dibasic Sequences in the Third Fibronectin-like Domain of L1Cell Adhesion Molecule (CAM) Facilitate Homomultimerization and Concomitant Integrin Recruitment  

Microsoft Academic Search

L1 is a multidomain transmembrane neural recognition molecule essential for neurohistogenesis. While moieties in the immunoglobulin-like domains of L1 have been implicated in both heterophilic and ho- mophilic binding, the function of the fibronectin (FN)-like repeats remains largely unresolved. Here, we demonstrate that the third FN-like repeat of L1 (FN3) spontaneously homomultimerizes to form trimeric and higher order complexes. Remarkably,

Steve Silletti; Fang Mei; Dean Sheppard; Anthony M. P. Montgomery



Inhibition of endogenous carcinoembryonic antigen (CEA) increases the apoptotic rate of colon cancer cells and inhibits metastatic tumor growth  

Microsoft Academic Search

It has been suggested that carcinoembryonic antigen (CEA) enhances metastatic seeding of colon cancer cells due to its homo-\\u000a and heterophilic binding properties. Our recent finding that endogenous CEA protects colon cancer cells against apoptosis\\u000a suggests a more complex role of CEA in cancer progression. In this study we compared the in vitro effects of endogenous CEA on tumor cell

Thomas Wirth; Edlyn Soeth; Frank Czubayko; Hartmut Juhl



Morphologic and cytochemical characteristics of Chinese striped-necked [corrected] turtle (Ocadia sinensis) blood cells and their hematologic and plasma biochemical reference values.  


Hematologic analyses are useful for the monitoring of animal health and diseases and for the differentiation of physiologic processes for clinicians and conservationists. In order to establish hematology reference values for the Chinese striped-necked [corrected] turtle (Ocadia sinensis) and to produce an accurate baseline of clinical laboratory data for O. sinensis with regard to sex and season, 50 (24 males and 26 females) adult captive individuals of O. sinensis were studied. Blood samples from the jugular veins of the turtles were collected in January, April, June, and November. Data were analyzed using repeated measures analysis of variance for significant (P < 0.05) variation by sex, season, and the interaction between sex and season. Significant sex differences were observed for the parameters of packed cell volume, eosinophil count, heterophils and monocytes ratio, total protein, albumin, uric acid, aspartate aminotransferase, alanine aminotransferase, triglycerides, cholesterol, and alkaline phosphatase. Marked seasonal variation was noted in all parameters except mean cell hemoglobin, monocytes and heterophils ratio, and creatinine. Differences between sexes and seasons were primarily associated with the reproductive cycle. Heterophils had a strong positive reaction and eosinophils had a moderate positive reaction to benzidine peroxidase stain. Thrombocytes had a positive reaction to periodic acid-Schiff stain. Surface morphologic study using scanning electron microscopy of blood cells showed that white blood cells of O. sinensis had no distinctive surface characteristics. PMID:19368243

Chung, Cheng-Shu; Cheng, Chiung-Hsiang; Chin, Shin-Chien; Lee, An-Hsing; Chi, Chau-Hwa



Hematologic parameters and hemoparasites of nonmigratory Canada geese (Branta canadensis) from Greensboro, North Carolina, USA.  


Large flocks of wild, nonmigratory Canada geese (Branta canadensis) have established permanent residence throughout the eastern United States and have become a public concern. Few studies have assessed the hematologic parameters for these populations, which could provide useful information for monitoring individual and population health of Canada geese. This study measured the hematologic parameters and detected the presence of hemoparasites from 146 wild, nonmigratory Canada geese in central North Carolina, USA, during their annual molt. The age class, sex, and weight of each bird were recorded at capture. Values for packed cell volume (PCV), estimated white blood cell count, white blood cell differentials, and heterophil: lymphocyte ratios were calculated for each bird. Adults and female geese had higher estimated white blood cell counts compared with juveniles and males, respectively. The PCV increased with weight and age class. Adult geese had higher percentages of heterophils and heterophil: lymphocyte ratios, whereas juvenile geese had higher percentages of lymphocytes. Relative eosinophil counts in adults increased with decreasing bird weight, and relative monocyte counts in juveniles increased with increasing weight. Three percent of geese were infected with species of Hemoproteus blood parasites. Atypical lymphocyte morphology, including pseudopods, split nuclei, and cytoplasmic granules, was observed in 5% of the birds. The hematologic values reported for adult and juvenile nonmigratory Canada geese in this study may serve as reference intervals for ecological studies and veterinary care of wild and captive Canada geese. PMID:24881149

Charles-Smith, Lauren E; Rutledge, M Elizabeth; Meek, Caroline J; Baine, Katherine; Massey, Elizabeth; Ellsaesser, Laura N; DePerno, Christopher S; Moorman, Christopher E; Degernes, Laurel A



Fluctuation of Spuriously Elevated Troponin I: A Case Report  

PubMed Central

Serum troponin is a useful laboratory study for the diagnosis of acute myocardial infarction. However, elevations can also be seen in a variety of other diseases processes. Falsely positive troponin values caused by interference with current troponin assays have been reported. We report a unique case that demonstrates the fluctuation of falsely elevated troponin correlating with hemoglobin, serving as a marker of heterophile antibody levels. A 74-year-old gentleman presented to our Emergency Department with a several-day history of increasing shortness of breath associated with a new-onset chest pain and a troponin I level of 77.28?ng/mL. Throughout his stay, fluctuations in measured troponin levels correlated strongly with fluctuations in hemoglobin levels. Several investigations confirmed false elevated troponin levels secondary to heterophile antibody interference. We conclude that hemoglobin trending in our patient represented a surrogate measure of his heterophile antibody titers with time and that fluctuations in these levels correlated with respective fluctuations in the falsely elevated troponin levels.

Ghali, Sam; Lewis, Keith; Kazan, Viviane; Altorok, Neezam; Taji, Jamil; Taleb, Mohammad; Lanka, Kiranmayee; Assaly, Ragheb



Neurite Fasciculation Mediated by Complexes of Axonin-1 and Ng Cell Adhesion Molecule  

PubMed Central

Neural cell adhesion molecules composed of immunoglobulin and fibronectin type III-like domains have been implicated in cell adhesion, neurite outgrowth, and fasciculation. Axonin-1 and Ng cell adhesion molecule (NgCAM), two molecules with predominantly axonal expression exhibit homophilic interactions across the extracellular space (axonin- 1/axonin-1 and NgCAM/NgCAM) and a heterophilic interaction (axonin-1–NgCAM) that occurs exclusively in the plane of the same membrane (cis-interaction). Using domain deletion mutants we localized the NgCAM homophilic binding in the Ig domains 1-4 whereas heterophilic binding to axonin-1 was localized in the Ig domains 2-4 and the third FnIII domain. The NgCAM–NgCAM interaction could be established simultaneously with the axonin-1–NgCAM interaction. In contrast, the axonin-1–NgCAM interaction excluded axonin-1/axonin-1 binding. These results and the examination of the coclustering of axonin-1 and NgCAM at cell contacts, suggest that intercellular contact is mediated by a symmetric axonin-12/NgCAM2 tetramer, in which homophilic NgCAM binding across the extracellular space occurs simultaneously with a cis-heterophilic interaction of axonin-1 and NgCAM. The enhanced neurite fasciculation after overexpression of NgCAM by adenoviral vectors indicates that NgCAM is the limiting component for the formation of the axonin-12/NgCAM2 complexes and, thus, neurite fasciculation in DRG neurons.

Kunz, Stefan; Spirig, Marianne; Ginsburg, Claudia; Buchstaller, Andrea; Berger, Philipp; Lanz, Rainer; Rader, Christoph; Vogt, Lorenz; Kunz, Beat; Sonderegger, Peter



Promotion of Spinal Cord Regeneration by Neural Stem Cell-Secreted Trimerized Cell Adhesion Molecule L1  

PubMed Central

The L1 cell adhesion molecule promotes neurite outgrowth and neuronal survival in homophilic and heterophilic interactions and enhances neurite outgrowth and neuronal survival homophilically, i.e. by self binding. We investigated whether exploitation of homophilic and possibly also heterophilic mechanisms of neural stem cells overexpressing the full-length transmembrane L1 and a secreted trimer engineered to express its extracellular domain would be more beneficial for functional recovery of the compression injured spinal cord of adult mice than stem cells overexpressing only full-length L1 or the parental, non-engineered cells. Here we report that stem cells expressing trimeric and full-length L1 are indeed more efficient in promoting locomotor recovery when compared to stem cells overexpressing only full-length L1 or the parental stem cells. The trimer expressing stem cells were also more efficient in reducing glial scar volume and expression of chondroitin sulfates and the chondroitin sulfate proteoglycan NG2. They were also more efficient in enhancing regrowth/sprouting and/or preservation of serotonergic axons, and remyelination and/or myelin sparing. Moreover, degeneration/dying back of corticospinal cord axons was prevented more by the trimer expressing stem cells. These results encourage the view that stem cells engineered to drive the beneficial functions of L1 via homophilic and heterophilic interactions are functionally optimized and may thus be of therapeutic value.

He, Xiaowen; Knepper, Michael; Ding, Cheng; Li, Jun; Castro, Suita; Siddiqui, Maham; Schachner, Melitta



The role of first line of defence mechanisms in the pathogenesis of cellulitis in broiler chickens: skin structural, physiological and cellular response factors.  


The present study examined several basic attributes of first-line defence mechanisms in the skin as potential factors that may explain the susceptibility of broiler chickens to cellulitis. The variables including structural characteristics of the skin, physicochemical properties and cellular responses to the challenge with pathogens were compared between two categories of chickens, a strain of fast-growing commercial broiler chickens (susceptible to cellulitis) and leghorn chickens (resistant to cellulitis). There were substantial differences between leghorns and broilers with regard to physiological characteristics of the skin. Broiler skin was more amenable to injury and the wound-healing process was slow. Compared with leghorns, the lesions resulting from sub-dermal challenge in broilers were more severe and disseminated over a larger area. Mobilization of phagocytic cells (heterophils and macrophages) in leghorns was brisk even in the areas distant from the site of infection, whereas only few heterophils were recruited in the skin of broilers. The functional competence of heterophils in broilers was inferior when compared with leghorns. Based on the present finding, the predisposition of broilers to cellulitis appears to be primarily associated with the inferior first line of defence of their skin. Broilers in commercial situations may be at higher risk to succumb to even minor infection and eventually develop cellulitis because: (1) structural weaknesses of the skin may predispose broilers to skin injury and thus the risk of skin infection by pathogens is increased; (2) broiler skin surface is more likely to provide a conducive environment for colonization of Escherichia coli; (3) in the event of infection, poor recruitment of phagocytic cells to the site of infection may readily lead to widespread colonization of the tissue by pathogens causing cellulitis and (4) poor functional quality of the phagocytic cells that are mobilized compromise the ability of the host to contain the spread of infection. PMID:16300661

Olkowski, A A; Wojnarowicz, C; Chirino-Trejo, M; Wurtz, B M; Kumor, L



Cytokine Signaling in Splenic Leukocytes from Vaccinated and Non-Vaccinated Chickens after Intravenous Infection with Salmonella Enteritidis  

PubMed Central

In order to design a new Salmonella enterica vaccine, one needs to understand how naive and immune chickens interact differently when exposed to S. enterica. In this study we therefore determined the immune response of vaccinated and non-vaccinated chickens after intravenous infection with Salmonella enterica serovar Enteritidis (S. Enteritidis). Using flow cytometry we showed that 4 days post infection (DPI), counts of CD4 and B-lymphocytes did not change, CD8 and ?? T-lymphocytes decreased and macrophages and heterophils increased in the spleen. When vaccinated and non-vaccinated chickens were compared, only macrophages and heterophils were found in significantly higher counts in the spleens of the non-vaccinated chickens. The non-vaccinated chickens also expressed higher anti-LPS antibodies than the vaccinated chickens. The expression of interleukin (IL)1?, IL6, IL8, IL18, LITAF, IFN? and iNOS did not exhibit any clear pattern in the cells sorted from the spleens of vaccinated or non-vaccinated chickens. Only IL17 and IL22 showed a differential expression in the CD4 T-lymphocytes of the vaccinated and non-vaccinated chickens at 4 DPI, both being expressed at a higher level in the non-vaccinated chickens. Due to a similar IFN? expression in the CD4 T-lymphocytes in both the vaccinated and non-vaccinated chickens, and a variable IL17 expression oscillating around IFN? expression levels, the IL17?IFN? ratio in CD4 T-lymphocytes was found to be central for the outcome of the immune response. When IL17 was expressed at higher levels than IFN? in the non-vaccinated chickens, the Th17 immune response with a higher macrophage and heterophil infiltration in the spleen dominated. However, when the expression of IL17 was lower than that of IFN? as in the vaccinated chickens, the Th1 response with a higher resistance to S. Enteritidis infection dominated.

Matulova, Marta; Stepanova, Hana; Sisak, Frantisek; Havlickova, Hana; Faldynova, Marcela; Kyrova, Kamila; Volf, Jiri; Rychlik, Ivan



p53 protein is absent from the serum of patients with lung cancer.  

PubMed Central

p53 protein, which accumulates intracellularly in over half of all human tumours, has also been reported to be present in the sera of patients with various malignancies, including lung cancer. Using a quantitative immunoassay, we measured p53 protein concentrations in 216 sera from 114 lung cancer patients of whom 75 provided matched lung tumour tissues, which were also assayed for p53 protein. p53 protein levels above the detection limit of 0.04 ng ml-1 were detected in only two sera from lung cancer patients (0.14 ng ml-1 and 0.27 ng ml-1), but not in any of 13 sera from non-malignant lung disease patients or in 100 sera from normal non-diseased individuals. The presence of these apparent traces of serum p53 protein concentrations could not be related either to the p53 protein expression status of the primary lung tumours or to the tumour stage, grade or histological type. By pretreating these two sera with anti-p53 antibody linked to solid phase, and by the addition of mouse serum to neutralise possible heterophilic antibodies, the signals arising from these sera were shown to be non-specific and possibly caused by heterophilic antibodies. We conclude that our data do not support previous reports of p53 protein in the sera of lung cancer patients. Since immunoassays are subject to numerous sources of interference in serum, including heterophilic antibodies, we suggest that the results of p53 protein analysis of serum specimens should be interpreted with caution.

Levesque, M. A.; D'Costa, M.; Diamandis, E. P.



Differential expression of Toll-like receptor mRNA in White Leghorn and indigenous chicken of India  

Microsoft Academic Search

In the present experiment, the expression profile of Toll-like receptor mRNA in indigenous and pure line chickens was studied.\\u000a The expression of TLR3, TLR4, TLR5 and TLR7 were quantified in heterophils of Aseel, Kadaknath, Naked neck, Dwarf and White\\u000a Leghorn lines by Quantitative Real-time PCR. White Leghorns expressed significantly (P?

Kannaki T. Ramasamy; Maddula R. Reddy; Dhanutha N. Raveendranathan; Shanmugam Murugesan; Rudra N. Chatterjee; Rajkumar Ullengala; Santosh Haunshi



Intestinal Cryptosporidium sp. infection in the Egyptian tortoise, Testudo kleinmanni.  


An adult Egyptian tortoise (Testudo kleinmanni) presented with clinical signs of enteritis and died 5 weeks after initiation of antibiotic therapy. Histological examination of the small intestine revealed heavy infection with Cryptosporidium sp.; over 80% of epithelial cells harboured the pathogen. No Cryptosporidium developmental stages were present in the stomach or the lungs. The intestinal lamina propria and mucosa were infiltrated by heterophils, lymphocytes and macrophages. The present study constitutes the first report of Cryptosporidium sp. infection in T. kleinmanni, and the first histological documentation of intestinal cryptosporidiosis in Chelonia. PMID:9925268

Graczyk, T K; Cranfield, M R; Mann, J; Strandberg, J D



Survival and physiologic response of Common Amakihi and Japanese White-eyes during simulated translocation  

USGS Publications Warehouse

We evaluated the effects of three translocation trials on Common Amakihi (Hemignathus virens) and Japanese White-eyes (Zosterops japonicus). Trial 1 involved capturing birds, transporting them on rough roads for 4 hr followed by holding in an aviary for 48 hr without overnight thermal support prior to release. Trial 2 involved capture, then holding in an aviary for 48 hr with overnight thermal support followed by transport for 4 hr prior to release. Trial 3 and 1 were identical except that overnight thermal support was provided during trial 3. We monitored survival, food consumption, weight change, and fecal production during captivity as well as changes in hematocrit, estimated total solids, heterophil to lymphocyte ratios, plasma uric acid, and creatinine phosphokinase (CPK) at capture and release. Survival was significantly lower for Amakihi during trial I (no thermal support). Birds that died lost significantly more weight than those that survived. Regardless of trial, birds responded to translocation by a combination of weight loss, anemia, hypoproteinemia, and elevated heterophil to lymphocyte ratio, uric acid, and CPK levels. The first 24 hr of captivity posed the greatest risk to birds regardless of whether transport or holding occurred first. Food consumption, fecal production, and weight all decreased at night, and overnight thermal support during holding was critical if ambient temperatures dipped to freezing. We recommend that if small passerines are to be held for > 12 hr, they be monitored individually for weight loss, food consumption, and fecal production.

Work, T. M.; Massey, J. G.; Johnson, L.; Dougill, S.; Banko, P. C.



Survival and physiologic response of common Amakihi and Japanese white-eyes during simulated translocation  

USGS Publications Warehouse

We evaluated the effects of three translocation trials on Common Amakihi (Hemignathus virens) and Japanese White-eyes (Zosterops japonicus). Trial 1 involved capturing birds, transporting them on rough roads for 4 hr followed by holding in an aviary for 48 hr without overnight thermal support prior to release. Trial 2 involved capture, then holding in an aviary for 48 hr with overnight thermal support followed by transport for 4 hr prior to release. Trial 3 and 1 were identical except that overnight thermal support was provided during trial 3. We monitored survival, food consumption, weight change, and fecal production during captivity as well as changes in hematocrit, estimated total solids, heterophil to lymphocyte ratios, plasma uric acid, and creatinine phosphokinase (CPK) at capture and release. Survival was significantly lower for Amakihi during trial 1 (no thermal support). Birds that died lost significantly more weight than those that survived. Regardless of trial, birds responded to translocation by a combination of weight loss, anemia, hypoproteinemia, and elevated heterophil to lymphocyte ratio, uric acid, and CPK levels. The first 24 hr of captivity posed the greatest risk to birds regardless of whether transport or holding occurred first. Food consumption, fecal production, and weight all decreased at night, and overnight thermal support during holding was critical if ambient temperatures dipped to freezing. We recommend that if small passerines are to be held for > 12 hr, they be monitored individually for weight loss, food consumption, and fecal production.

Work, T. M.; Massey, J. G.; Johnson, L.; Dougill, S.; Banko, P. C.



Prevalence and pathology of the nematode Heterakis gallinarum, the trematode Paratanaisia bragai, and the protozoan Histomonas meleagridis in the turkey, Meleagris gallopavo.  


The prevalence of infection and associated pathology induced by two helminth and one protozoan species infecting Brazilian turkeys are reported. The intestinal nematode Heterakis gallinarum appeared with a prevalence of 70% in the infected birds, without gross lesions when not associated to the protozoan Histomonas meleagridis. Histological findings in the ceca were represented by the presence of H. gallinarum worms, intense chronic diffuse inflammatory processes with mononuclear and polymorphonuclear (heterophils) leucocyte infiltrations. The prevalence of the protozoan H. meleagridis associated to H. gallinarum was of 2.5% and microscopic examination revealed a severe inflammatory process in the liver and cecum with the presence of small clear areas with round eosinophilic parasites. Gross lesions were absent in turkeys infected with the renal digenetic trematode Paratanaisia bragai; the parasite was prevalent in 20% of the cases and cross-sections of the kidneys showed a remarkable distension of the collecting ducts with several worms in the lumen. The walls of the ducts presented a discrete heterophilic infiltrate among mononuclear cells. PMID:17072483

Brener, Beatriz; Tortelly, Rogério; Menezes, Rodrigo Caldas; Muniz-Pereira, Luís C; Pinto, Roberto Magalhães



Neuronal synapse interaction reconstituted between live cells and supported lipid bilayers  

PubMed Central

In the nervous system, homophilic and heterophilic adhesion molecules participate in the induction and differentiation of presynaptic transmitter release sites. We focus on the heterophilic interaction between postsynaptic neuroligin-1 (Nlg) and presynaptic ?-neurexin (Nrx). Nlg has previously been shown to trigger presynaptic differentiation in a Nrx-expressing axon even when presented on a non-neuronal cell or on beads coated with lipid bilayers. We have now developed a new method to measure single molecule and ensemble distribution of Nrx and Nlg at the contact site between a non-neuronal Nrx-expressing cell and a flat supported glycosylphosphoinositol–neuroligin-1 (GPI-Nlg) lipid bilayer and relate them to adhesion as measured by cell migration and gravity dissociation. We find that within minutes after cell-bilayer contact, Nrx accumulates at the contact site and the contact area is expanded. The strength of cell-bilayer adhesion depends on the morphology of Nrx accumulation, with the focal concentration strengthening adhesion. The results suggest that Nlg-Nrx interaction rapidly establishes a weak, but specific, adhesion between dynamic pre- and postsynaptic processes, which may ultimately require additional molecules for synapse stabilization.

Pautot, Sophie; Lee, Hanson; Isacoff, Ehud Y; Groves, Jay T



West Nile Virus in Farmed Alligators  

PubMed Central

Seven alligators were submitted to the Tifton Veterinary Diagnostic and Investigational Laboratory for necropsy during two epizootics in the fall of 2001 and 2002. The alligators were raised in temperature-controlled buildings and fed a diet of horsemeat supplemented with vitamins and minerals. Histologic findings in the juvenile alligators were multiorgan necrosis, heterophilic granulomas, and heterophilic perivasculitis and were most indicative of septicemia or bacteremia. Histologic findings in a hatchling alligator were random foci of necrosis in multiple organs and mononuclear perivascular encephalitis, indicative of a viral cause. West Nile virus was isolated from submissions in 2002. Reverse transcription-polymerase chain reaction (RT-PCR) results on all submitted case samples were positive for West Nile virus for one of four cases associated with the 2001 epizootic and three of three cases associated with the 2002 epizootic. RT-PCR analysis was positive for West Nile virus in the horsemeat collected during the 2002 outbreak but negative in the horsemeat collected after the outbreak.

Mauel, Michael J.; Baldwin, Charles; Burtle, Gary; Ingram, Dallas; Hines, Murray E.; Frazier, Kendal S.



Extracellular architecture of the SYG-1/SYG-2 adhesion complex instructs synaptogenesis.  


SYG-1 and SYG-2 are multipurpose cell adhesion molecules (CAMs) that have evolved across all major animal taxa to participate in diverse physiological functions, ranging from synapse formation to formation of the kidney filtration barrier. In the crystal structures of several SYG-1 and SYG-2 orthologs and their complexes, we find that SYG-1 orthologs homodimerize through a common, bispecific interface that similarly mediates an unusual orthogonal docking geometry in the heterophilic SYG-1/SYG-2 complex. C. elegans SYG-1's specification of proper synapse formation in vivo closely correlates with the heterophilic complex affinity, which appears to be tuned for optimal function. Furthermore, replacement of the interacting domains of SYG-1 and SYG-2 with those from CAM complexes that assume alternative docking geometries or the introduction of segmental flexibility compromised synaptic function. These results suggest that SYG extracellular complexes do not simply act as "molecular velcro" and that their distinct structural features are important in instructing synaptogenesis. PAPERFLICK: PMID:24485456

Özkan, Engin; Chia, Poh Hui; Wang, Ruiqi Rachel; Goriatcheva, Natalia; Borek, Dominika; Otwinowski, Zbyszek; Walz, Thomas; Shen, Kang; Garcia, K Christopher



Effects of humic acid on broiler chickens.  


In view of the alleged effect of humic acid (HA) on growth plate arthrosis in humans, we sought to find if poultry tibial dyschondroplasia (TD) is caused by HA that can occur as a ground water contaminant. In 2 separate trials, broiler chickens were fed different concentrations of HA added to their diet for 4 and 5 wk. The effects of these treatments were measured by general health indices such as growth, feed conversion ratio, relative organ weights, blood differential count, serum chemistry, TD index, and bone biomechanical strength. Humic acid treatment decreased BW but appeared to improve feed conversion ratio. There was no effect on TD index or bone biomechanical strength in HA-treated birds compared with controls. There was no toxic effect of HA that was evident by the absence of any dramatic change in relative organ weights or other telltale signs of serum clinical chemistry that would suggest liver, muscle, or kidney dysfunction. Red blood cell, white blood cell, monocyte, and hematocrit values were not affected, but there was a decrease in blood heterophil counts and heterophil to lymphocyte ratio, which was significant in 4-wk HA-treated birds. Overall these results show that HA slows down growth, but it does not have any adverse health effects on chickens. PMID:16553268

Rath, N C; Huff, W E; Huff, G R



Expression and purification of an Epstein-Barr virus encoded 23-kDa protein and characterization of its immunological properties.  


Serodiagnosis of Epstein-Barr virus (EBV) infection is currently based on the detection of antibodies to distinct EBV antigens by immunofluorescence and enzyme-linked immunosorbent assay-based tests, or in part on the detection of heterophile antibodies by the Paul-Bunnell-Davidson heterophile assay. In the past few years, the specificity and the sensitivity of serodiagnostic assay systems has been improved considerably by the use of purified recombinant EBV antigens. Screening of EBV-positive sera for antigenic reactivities by immunoprecipitation with extracts of EBV-positive cells revealed a 23-kDa protein (p23) that was recognized by antibodies from all EBV carriers tested. Open reading frame BLRF2 was identified as the coding region for this protein. After cloning and high-level expression of the BLRF2 open reading frame as DHFR fusion protein in Escherichia coli, the recombinant protein was purified to near homogeneity with the help of continuous elution electrophoresis. Sera from both EBV-positive and -negative donors were screened by immunoblot analysis and enzyme-linked immunosorbent assay for IgM and IgG antibodies against the EBV-encoded protein p23. Since anti-p23 antibodies were not detectable in 30 of 30 EBV-negative sera, and 294 of 302 EBV-positive sera had either IgM and/or IgG antibody responses to this protein, recombinant p23 seems to be a useful diagnostic marker for EBV-infection. PMID:8919825

Reischl, U; Gerdes, C; Motz, M; Wolf, H



Effect of inositol and phytases on hematological indices and ?-1 acid glycoprotein levels in laying hens fed phosphorus-deficient corn-soybean meal-based diets.  


The effects of feeding low nonphytate phosphorus (NPP) corn-soybean meal-based diets supplemented with myo-inositol at 0.1%, or with phytase B at 1,300 acid phosphatase units/kg, or with phytase B enriched in 6-phytase A at 300 phytase units/kg on the hematological indices and the ?-1 acid glycoprotein (AGP) concentrations in the blood of Bovans Brown laying hens were investigated. The experimental design comprised also a negative control diet and an internal control diet that had the NPP content adjusted by the addition of 0.304 g of monocalcium phosphate per kg to reach the NPP level similar to that resulting from the combined action of both phytases. A total of sixty 50-wk-old hens were randomly assigned to the dietary treatments with 12 cage replicates of 1 hen, and fed the experimental diets until wk 62, when the blood samples were taken and analyzed for basic hematological indices and for AGP concentrations in sera. The hematological indices from all the experimental groups remained in a normal range; nevertheless, the statistically significant effects of diet on hemoglobin concentration (P = 0.003), erythrocyte counts (P = 0.035), the percentage of lymphocytes (P = 0.020), heterophils (P = 0.002), eosinophils (P = 0.023), and basophils (P = 0.001) in the leucocyte population, as well as on the heterophil to lymphocyte ratio (P = 0.003), were observed. The highest erythrocyte counts were characteristic for hens fed the diet supplemented with both phytase A and phytase B. The highest heterophil to lymphocyte ratios were found in blood of hens fed the diet supplemented with phytase B, whereas the highest basophil percentages and the highest AGP concentrations occurred in birds fed the negative control diet. A highly significant correlation was observed between AGP concentrations in sera and BW losses determined previously. The results indicate that the low-NPP corn soybean meal-based diets increased acute phase protein level in laying hens. Phytase B alone, and particularly in combination with phytase A, acted as a potent mediator of the response, whereas supplementary myo-inositol did not. PMID:23243248

Zy?a, K; Grabacka, M; Pierzchalska, M; Duli?ski, R; Starzy?ska-Janiszewska, A



Epstein-Barr Virus Infection with Acute Pancreatitis Associated with Cholestatic Hepatitis  

PubMed Central

Infection-induced acute hepatitis complicated with acute pancreatitis is associated with hepatitis A virus, hepatitis B virus or hepatitis E virus. Although rare, Epstein-Barr virus (EBV) infection should be considered also in the differential diagnosis if the patient has acute hepatitis combined with pancreatitis. We report a case of EBV infection with cholestatic hepatitis and pancreatitis with review of literature. An 11-year-old female was admitted due to 1-day history of abdominal pain and vomiting without any clinical symptoms of infectious mononucleosis. Diagnosis of reactivated EBV infection was made by the positive result of viral capsid antigen (VCA) IgM, VCA IgG, Epstein-Barr nuclear antigen and heterophile antibody test. We performed serologic tests and magnetic resonance cholangiopancreatography to exclude other viral or bacterial infection, autoimmune disorder, and structural problems. The patient's symptoms recovered rapidly and blood chemistry returned to normal with conservative treatment similar to previously reported cases.

Kang, Seok-Jin; Yoon, Ka-Hyun



Missing links in multiple sclerosis etiology. A working connecting hypothesis.  


The etiology of multiple sclerosis is still elusive despite an extended patchwork of mechanistic events has been accumulated. In this article, are tentatively identified from scattered literature sources new factors that may link well known characteristic of MS such as the central alteration of BBR selectivity, its association with EBV status and its biased distribution of the globe more comprehensively. The hypothesis proposes that the concomitant important rise in some heterophilic antibodies (anti Neu5Gc) which activate BBB endothelial cells and in the frequency of anti EBV committed T cells and of memory B infected cells with EBV contemporary to EBV infection play a major role in MS etiology. In addition, the hypothesis proposes new possible explanations for the elevated risk of MS in specific geographical area. PMID:23466062

Soulillou, Jean-Paul



Circular Epstein-Barr virus genomes of reduced size in a human lymphoid cell line of infectious mononucleosis origin.  

PubMed Central

Circular Epstein-Barr virus (EBV) DNA molecules have been purified and characterized from a human lymphoid cell line derived from a case of heterophile antibody-positive, blood transfusion-induced infectious mononucleosis, 883L. The circular EBV DNA in three cell lines obtained by transformation of human umbilical cord blood leukocytes with a strain of EBV originally derived from 883L was also studied. As estimated from sedimentation velocity data and electron microscopy, the circular EBV DNA molecules are 10 to 15% smaller than either the circular EBV DNA previously found intracellularly in several other types of EBV-transformed cells or the linear EBV DNA present extracellularly in virus particles. In addition, the EBV-transformed cord blood cell lines studied here differed from other EBV-transformed cells in that integrated virus DNA sequences could not be detected. Images

Adams, A; Bjursell, G; Kaschka-Dierich, C; Lindahl, T



Sarcocystis-associated encephalitis in chickens.  


Sarcocystis-associated encephalitis was diagnosed in a backyard chicken flock that had nervous manifestations. The main histopathologic lesion was necrotizing encephalitis characterized by a large focal area of necrosis infiltrated and surrounded by mononuclear cells, heterophils, and multinucleated giant cells. Schizonts and merozoites were observed in the lesion. Immunohistochemical staining of the brain lesion revealed positive reaction to Sarcocystis antiserum. The ultrastructural characteristics of the parasite were typical of Sarcocystis, including the presence of a nucleus, a conoid, numerous micronemes, and lack of rhoptries. Medication with amprolium and sulfamethazine or with chlortetracycline was not effective in controlling the mortality. Trapping of opossums on the farm and relocating the chickens to clean, new premises seemed to reduce mortality from this infection. PMID:7677669

Mutalib, A; Keirs, R; Maslin, W; Topper, M; Dubey, J P



Ultrastructural pathology of Bordetella avium infection in turkeys.  


One-day-old turkeys were infected intranasally with Bordetella avium, and tracheas were examined by scanning and transmission electron microscopy at 1 to 5 weeks post-inoculation (PI). The predominant ultrastructural lesions were progressive loss of ciliated epithelium with replacement by nonciliated cells, bacterial colonization of ciliated cells, membrane-bound crystalline inclusions in cytoplasma of epithelial cells, depletion of mucous granules, and distortion of tracheal rings and the mucosal surface. Tracheal surface exudates consisted of mucus, necrotic cells, heterophils, and fibrin. Ciliated cells were replaced by immature cuboidal cells characterized by abundant rough endoplasmic reticulum with small numbers of electron-dense mucous granules in the apical cytoplasm. Bacterial surfaces were rough and contained numerous pleomorphic, knob-like structures, 20-50 nm in diameter. Other changes included enlarged mucosal gland openings, cell extrusion marks, pleomorphic microvilli, and cells with small numbers of short cilia. PMID:3672806

Arp, L H; Fagerland, J A



Effect of exposure to operant-controlled microwaves on certain blood and immunological parameters in the young chick  

SciTech Connect

Twenty-two 1-wk-old broiler chicks (Gallus domesticus) were housed at 16 C and operantly conditioned to activate either a 250-W infrared bulb (control) or a microwave generator delivering 13 mW/cm2 (treated). Plasma corticosterone concentration did not differ between groups (P greater than .05) at 4 wk of age. At that time the birds were killed, and post-mortem examination revealed no treatment differences in gross morphology of the chicks or in weights of spleen and bursa of Fabricius (P greater than .05). Histological study of comparable segments of spleen, bursa, adrenal, and thyroid tissue did not show differences in any of the chosen parameters (P greater than .05). Heterophil:lymphocyte ratios, packed cell volume, and total plasma protein content were similar between groups (P greater than .05). These results suggest that operant exposure to low density microwave radiation did not result in stress or immunological disturbances.

Braithwaite, L.A.; Morrison, W.D.; Bate, L.; Otten, L.; Hunter, B.; Pei, D.C. (Department of Animal and Poultry Science, University of Guelph, Ontario (Canada))



Synapses: Sites of Cell Recognition, Adhesion, and Functional Specification  

PubMed Central

Synapses are specialized adhesive contacts characteristic of many types of cell-cell interactions involving neurons, immune cells, epithelial cells, and even pathogens and host cells. Cell-cell adhesion is mediated by structurally diverse classes of cell-surface glycoproteins, which form homophilic or heterophilic interactions across the intercellular space. Adhesion proteins bind to a cytoplasmic network of scaffolding proteins, regulators of the actin cytoskeleton, and signal transduction pathways that control the structural and functional organization of synapses. The themes of this review are to compare the organization of synapses in different cell types and to understand how different classes of cell adhesion proteins and cytoplasmic protein networks specify the assembly of functionally distinct synapses in different cell contexts.

Yamada, Soichiro; Nelson, W. James



Differences in response of chickens from two genetic lines to diisopropyl phosphorofluoridate.  


Neurotoxicity of diisopropyl phosphorofluoridate (DFP) was examined at 85 weeks of age in hens of two lines selected for high (HA) and low (LA) antibody response to sheep erythrocytes. DFP was administered by subcutaneous injection in doses of 0.25, 0.50 and 1.00 mg/kg and hens were observed for cholinergic signs at 30 min and for delayed neuropathy 8 to 14 days post-administration. Toxicity to DFP increased in severity with the dose and genetic differences were present because hens of line HA were more sensitive to DFP than were those of line LA. HA hens also had lower A-esterase activities and higher heterophil-to-lymphocyte ratios. No line x treatment interaction was evident, however, for activities of neurotoxic esterase or brain cholinesterase measured 24 hr after DFP administration. PMID:2549474

Dunnington, E A; Siegel, P B; Ehrich, M



Crystal structure of murine coronavirus receptor sCEACAM1a[1,4],a member of the carcinoembtyonic antigen family  

SciTech Connect

CEACAM1 is a member of the carcinoembryonic antigen (CEA) family. Isoforms of murine CEACAM1 serve as receptors for mouse hepatitis virus (MHV), a murine coronavirus. Here we report the crystal structure of soluble murine sCEACAM1a[1,4], which is composed of two Ig-like domains and has MHV neutralizing activity. Its N-terminal domain has a uniquely folded CC' loop that encompasses key virus-binding residues. This is the first atomic structure of any member of the CEA family, and provides a prototypic architecture for functional exploration of CEA family members. We discuss the structural basis of virus receptor activities of murine CEACAM1 proteins, binding of Neisseria to human CEACAM1, and other homophilic and heterophilic interactions of CEA family members.

Tan, K.; Zelus, B. D.; Meijers, R.; Liu, J.-H.; Bergelson, J. M.; Zhang, R.; Duke, N.; Joachimiak, A.; Holmes, K. V.; Wang, J.-H.; Biosciences Division; Dana-Farber Cancer Inst.; Harvard Medical School; Univ. of Colorado Health Science Center; Univ. of Pennsylvania School of Medicine



Salpingitis in geese associated with Mycoplasma sp. strain 1220.  


An outbreak of disease in a White Rhine laying goose flock was characterized by increased water uptake, increased mortality, production of eggs with abnormal shells, a 25% drop in egg production and 40% embryo mortality. Affected dead or sacrificed birds had sero-fibrinogranulocytic peritonitis and salpingitis, infiltration of the lamina propria in the uterus and heterophil granulocytes in the isthmus and magnum of the oviduct. Mycoplasmas, mainly identified as Mycoplasma sp. strain 1220, were isolated from the airsac, liver, ovary, magnum and peritoneum of some affected geese. Strain 1220 was originally isolated from a Hungarian gander with phallus inflammation and, according to detailed biochemical and serological examinations, it is expected to represent a new avian species within the genus Mycoplasma. PMID:19468942

Dobos-Kovács, Mihály; Varga, Zsuzsanna; Czifra, György; Stipkovits, László



Salt gland adenitis as only cause of stranding of loggerhead sea turtles Caretta caretta.  


The present study describes pathological and microbiological findings in 9 stranded loggerhead sea turtles Caretta caretta, whose only observed lesion was bilateral purulent salt gland adenitis. Histological lesions ranged from the presence of abundant eosinophilic material associated with bacterial colonies in the lumen of the central ducts of the glandular lobules to the destruction of the glandular tissue and presence of abundant eosinophilic material composed of heterophils and cell debris, lined by multinucleated giant cells. Aeromonas hydrophila, Staphylococcus sp., and Vibrio alginolyticus were the bacteria most frequently isolated. Plasma concentrations of sodium and chloride and plasma osmolality from 2 turtles suffering from salt gland adenitis were, respectively 45.7, 69.2, and 45.7% higher than the mean value for healthy turtles. These cases suggest that failure to maintain homeostasis due to severe lesions in the salt glands can cause stranding and/or death of loggerhead sea turtles. PMID:21848124

Orós, J; Camacho, M; Calabuig, P; Arencibia, A



NCAM-induced intracellular signaling revisited.  


The neural cell adhesion molecule (NCAM) plays a crucial role in neuronal development, synaptic plasticity, and regeneration. NCAM works as "smart glue" that not only mediates cell-cell adhesion but also induces activation of a complex network of intracellular signaling cascades on homophilic or heterophilic binding. Stimulation of NCAM by homophilic interactions induces neuronal differentiation through activation of a number of signaling molecules, including the fibroblast growth factor receptor, non-receptor kinases Fyn and focal adhesion kinase, growth-associated protein-43, the mitogen-activated protein kinase pathway, intracellular Ca(2+), and protein kinases A, C, and G. This review presents and discusses the current knowledge in the area of NCAM signaling with a focus on the events involved in NCAM-mediated neurite outgrowth. PMID:17975827

Ditlevsen, Dorte Kornerup; Povlsen, Gro Klitgaard; Berezin, Vladimir; Bock, Elisabeth



Granulopoiesis in tadpoles of Rana esculenta. Survey of the organs involved.  

PubMed Central

The topography and nature of granulopoietic tissues appearing during the larval period of Rana esculenta have been investigated using light and electron microscopy. The formation of eosinophils and heterophils occurred in the interstices of the pronephros and opisthonephros, in the mesenchymal sheath surrounding the pronephric duct, as well as in the mesentery and mesenchymal coat of the bile duct and hepatic ducts. Developing basophils were very rare, being detectable only in the stroma of the trunk kidneys. The pronephric interstices additionally contained differentiating lymphocytes, the opisthonephric interstices differentiating lymphocytes and erythrocytes. In contrast, the mesenchymal sheath of the pronephric duct as well as the mesentery and the mesenchymatous coat of the bile duct and hepatic ducts were solely granulopoietic. Images Fig. 1 Figs. 2-3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 Fig. 10 Fig. 11 Fig. 12

Frank, G



Hematology, plasma biochemistry, and antibodies to select viruses in wild-caught Eastern massasauga rattlesnakes (Sistrurus catenatus catenatus) from Illinois.  


During the 2004 field season, blood was collected from Eastern massasauga rattlesnakes (Sistrurus catenatus catenatus) in the Carlyle Lake (Carlyle, Illinois, USA) and Allerton Park (Monticello, Illinois, USA) populations to derive baseline complete blood count and plasma biochemistry data and to assess the prevalence of antibodies to West Nile virus (WNV) and ophidian paramyxovirus (OPMV). Massasaugas were located for sampling through visual encounter surveys. Body weight, snout-vent length, total protein, globulins, sodium, and potassium were normally distributed among the survey population. Aspartate aminotransferase, creatine kinase, albumin, calcium, uric acid, white blood cell count, heterophils, lymphocytes, monocytes, eosinophils, and basophils were non-normally distributed within these animals. Female snakes had significantly shorter tail lengths; lower blood glucose, packed cell volumes, and absolute azurophil counts; and higher plasma calcium and phosphorus concentrations than did males. None of the snakes tested (n=21) were seropositive for WNV, whereas all (n=20) were seropositive for OPMV. PMID:16699153

Allender, Matthew C; Mitchell, Mark A; Phillips, Christopher A; Gruszynski, Karen; Beasley, Val R



CEA-Related CAMs.  


The carcinoembryonic antigen (CEA) family comprises a large number of cellular surface molecules, the CEA-related cell adhesion molecules (CEACAMs), which belong to the Ig superfamily. CEACAMs exhibit a complex expression pattern in normal and malignant tissues. The majority of the CEACAMs are cellular adhesion molecules that are involved in a great variety of distinct cellular processes, for example in the integration of cellular responses through homo- and heterophilic adhesion and interaction with a broad selection of signal regulatory proteins, i.e., integrins or cytoskeletal components and tyrosine kinases. Moreover, expression of CEACAMs affects tumor growth, angiogenesis, cellular differentiation, immune responses, and they serve as receptors for commensal and pathogenic microbes. Recently, new insights into CEACAM structure and function became available, providing further elucidation of their kaleidoscopic functions. PMID:20455097

Horst, A K; Wagener, C



TgrC1 mediates cell-cell adhesion by interacting with TgrB1 via mutual IPT/TIG domains during development of Dictyostelium discoideum.  


Cell-cell adhesion plays crucial roles in cell differentiation and morphogenesis during development of Dictyostelium discoideum. The heterophilic adhesion protein TgrC1 (Tgr is transmembrane, IPT, IG, E-set, repeat protein) is expressed during cell aggregation, and disruption of the tgrC1 gene results in the arrest of development at the loose aggregate stage. We have used far-Western blotting coupled with MS to identify TgrB1 as the heterophilic binding partner of TgrC1. Co-immunoprecipitation and pull-down studies showed that TgrB1 and TgrC1 are capable of binding with each other in solution. TgrB1 and TgrC1 are encoded by a pair of adjacent genes which share a common promoter. Both TgrB1 and TgrC1 are type I transmembrane proteins, which contain three extracellular IPT/TIG (immunoglobulin, plexin, transcription factor-like/transcription factor immunoglobulin) domains. Antibodies raised against TgrB1 inhibit cell reassociation at the post-aggregation stage of development and block fruiting body formation. Ectopic expression of TgrB1 and TgrC1 driven by the actin15 promoter leads to heterotypic cell aggregation of vegetative cells. Using recombinant proteins that cover different portions of TgrB1 and TgrC1 in binding assays, we have mapped the cell-binding regions in these two proteins to Lys(537)-Ala(783) in TgrB1 and Ile(336)-Val(360) in TgrC1, corresponding to their respective TIG3 and TIG2 domain. PMID:23477311

Chen, Gong; Wang, Jun; Xu, Xiaoqun; Wu, Xiangfu; Piao, Ruihan; Siu, Chi-Hung



Immune Response of Broiler Chickens Fed Diets Supplemented with Different Level of Chromium Methionine under Heat Stress Conditions  

PubMed Central

The objectives of this study were to investigate the immune responses of broiler chickens fed diets supplemented with different level of chromium methionine (CrMet) in heat stress (HS) condition. Two hundred and eighty eight male broiler chickens (Ross 308) were allocated to four treatment groups (supplementation with 0, 200, 400 or 800 ppb Cr in the form of CrMet) in a completely randomized design. The experiment was conducted at heat stressed condition and all birds were kept under temperature of 33±2°C. Antibody titers against Newcastle disease virus (NDV) and infectious bronchitis virus (IBV), heterophil to lymphocyte ratios (H/L), and concentration of plasma cortisol (CPC) were measured at 21 and 42 d. At 42 days of age two birds were chosen randomly from each replicate, slaughtered, spleen and bursa of Fabricius were collected, weighed and expressed as a percentage of live body weight. Antibody titers against NDV and IBV at 21 and 42 days of age in broiler fed supplemental CrMet were higher than in broiler chickens fed control diet (p<0.05). CPC level in broiler chickens fed CrMet were significantly (p<0.05) decreased. Increases in lymphocyte counts and consequently a decrease in heterophil to lymphocyte ratios in broiler chickens fed 800 ppb Cr were observed at 21 and 42 d. Supplementation with CrMet had no significant effect on lymphoid organs of broilers. The results suggest that dietary CrMet supplementation at a level of 800 ppb can improve some immune responses of broiler chickens under heat stress conditions.

Ebrahimzadeh, S. K.; Farhoomand, P.; Noori, K.



Capability of different non-nutritive feed additives on improving productive and physiological traits of broiler chicks fed diets with or without aflatoxin during the first 3 weeks of life.  


An experiment was conducted to determine whether some non-nutritive feed additives (NNFA) could block the adverse effects of aflatoxin (AF) on growth performance and physiological parameters of Cobb broilers throughout the period from 1 to 21 day of age. There were eight treatments consisting of two levels of AF at 0 and 200 ppb and four NNFA within each AF level. These additives included mannan oligosaccharides (MOS) at 2 g/kg diet, hydrated sodium calcium aluminosilicate (HSCAS) at 2 g/kg diet and Lactobacillus acidophilus (Lac) at 2 g/kg diet. At 21 day of age, five chickens of each treatment were slaughtered to study dressing percentage and relative weight of inner organs and glands. AF had a significant negative effect on body weight gain (BWG), and feed intake, while impairing feed conversion ratio (FCR). Aflatoxin significantly increased percentage liver, lymphocyte (%), monocyte (%), serum triglyceride level, and the aspartate aminotransferase (AST), and alanine aminotransferase (ALT), concentrations while decreasing dressing percentage, intestinal percentage, white blood cells (WBCs), red blood cells (RBCs), haemoglobin (Hgb), packed cell volume (PCV), heterophil (%), heterophil/lymphocyte ratio, total serum protein and serum albumin. Aflatoxin adversely affected the morphology of the liver, bursa and the thymus. There was a significant interaction between AF and NNFA on the relative weights of liver, heart and intestine. Lac completely blocked the negative effects of AF on the percentage liver and the heart and partially on the intestine. In conclusion, Lac was most effective in reversing the adverse effects of AF on growth and FCR and on the percentage, functions and morphology of the liver. Hydrated sodium calcium aluminosilicate also improved the economic traits of broilers but was less effective than Lac and more effective than MOS. PMID:23050696

Attia, Y A; Allakany, H F; Abd Al-Hamid, A E; Al-Saffar, A A; Hassan, R A; Mohamed, N A



Molecular interactions between desmosomal cadherins.  

PubMed Central

Desmocollins (Dscs) and desmogleins (Dsgs) are cell-adhesion molecules involved in the formation of desmosome cell-cell junctions and share structural similarities to classical cadherins such as E-cadherin. In order to identify and provide quantitative information on the types of protein-protein interactions displayed by the type 2 isoforms and investigate the role of Ca(2+) in this process, we have developed an Escherichia coli expression system to generate recombinant proteins containing the first two extracellular domains, namely Dsg2(1-2) and Dsc2(1-2). Analytical ultracentrifugation, chemical cross-linking, CD, fluorescence and BIAcore have been used to provide the first direct evidence of Ca(2+) binding to desmosomal cadherins. These studies suggest that Dsc2(1-2) not only exhibits homophilic interactions in solution, but can also form heterophilic interactions with Dsg2(1-2). The latter, on the other hand, shows much weaker homophilic association. Our results further demonstrate that heterophilic interactions are Ca(2+)-dependent, whereas the Ca(2+)-dependence of homophilic association is less clear. Our data indicate that the functional properties of Dsc2(1-2) are more similar to those of classical cadherins, consistent with the observation that Dsc shares a higher level of sequence homology with classical cadherins than does Dsg. In addition to corroborating the conclusions of previously reported transfection studies which suggest the formation of lateral heterodimers and homodimers, our results also provide direct quantitative information on the strength of these interactions which are essential for understanding the adhesion mechanism.

Syed, Shabih-e-Hassnain; Trinnaman, Brian; Martin, Stephen; Major, Sarah; Hutchinson, Jon; Magee, Anthony I



Oral exposure of broiler breeder hens to extra thyroxine modulates early adaptive immune responses in progeny chicks.  


Based on the findings of a recent study suggesting a decreased cold-induced ascites incidence in broiler progeny from hyperthyroid (HYPER) breeder hens, and a controversy on the effects of hyperthyroidism on immunocompetence, the present study was conducted to determine the probable adverse effect of induced maternal hyperthyroidism on immune function in progeny chicks. Breeder hens (n = 88) were randomly allotted to the control or HYPER groups and received common or thyroxine (T4)-added (1 mg/L) water, respectively. The hens were artificially inseminated, and hatching eggs (n = 924) were incubated. Thereafter, the male hatchlings (n = 288) were reared for 42 d, and several cellular and humoral immune responses were evaluated at standard or low ambient temperature. Prevaccination antibody titers to Newcastle disease, infectious bronchitis, and infectious bursal disease virus were higher in HYPER chicks during 1 wk of age, although not different in their dams. For primary response to SRBC administered at 7 d of age, HYPER chicks recorded higher total, IgM (d 14), and IgG (d 21) anti-SRBC antibody titers. Higher cutaneous basophilic hypersensitivity response in HYPER chicks (d 10) was not observed at 35 d of age. Carbon clearance assay showed no difference, but in vitro lymphoproliferative response to concanavalin A was higher in 19-d-old HYPER chicks, independent of temperature treatment. An increase in lymphocyte percentage coincided with a decreased heterophil percentage and heterophil to lymphocyte ratio (d 14) in the HYPER group. The weight of lymphoid organs in progeny was not influenced by the oral exposure of dams to extra T4. Independent of T4 treatment, cold exposure was generally associated with decreased immune functions at early stages. The data suggested that oral exposure of broiler breeder hens to 1 mg/L of T4 not only had no adverse effect on immune function, but also modulated early adaptive immune responses in progeny chicks for which the causal mechanisms remain to be unraveled. PMID:23472027

Akhlaghi, A; Zamiri, M J; Jafari Ahangari, Y; Atashi, H; Ansari Pirsaraei, Z; Deldar, H; Eghbalian, A N; Akhlaghi, A A; Navidshad, B; Yussefi Kelarikolaei, K; Hashemi, S R



Hematology of healthy Florida manatees (Trichechus manatus)  

USGS Publications Warehouse

Background: Hematologic analysis is an important tool in evaluating the general health status of free-ranging manatees and in the diagnosis and monitoring of rehabilitating animals. Objectives: The purpose of this study was to evaluate diagnostically important hematologic analytes in healthy manatees (Trichechus manatus) and to assess variations with respect to location (free ranging vs captive), age class (small calves, large calves, subadults, and adults), and gender. Methods: Blood was collected from 55 free-ranging and 63 captive healthy manatees. Most analytes were measured using a CELL-DYN 3500R; automated reticulocytes were measured with an ADVIA 120. Standard manual methods were used for differential leukocyte counts, reticulocyte and Heinz body counts, and plasma protein and fibrinogen concentrations. Results: Rouleaux, slight polychromasia, stomatocytosis, and low numbers of schistocytes and nucleated RBCs (NRBCs) were seen often in stained blood films. Manual reticulocyte counts were higher than automated reticulocyte counts. Heinz bodies were present in erythrocytes of most manatees. Compared with free-ranging manatees, captive animals had slightly lower MCV, MCH, and eosinophil counts and slightly higher heterophil and NRBC counts, and fibrinogen concentration. Total leukocyte, heterophil, and monocyte counts tended to be lower in adults than in younger animals. Small calves tended to have higher reticulocyte counts and NRBC counts than older animals. Conclusions: Hematologic findings were generally similar between captive and free-ranging manatees. Higher manual reticulocyte counts suggest the ADVIA detects only reticulocytes containing large amounts of RNA. Higher reticulocyte and NRBC counts in young calves probably reflect an increased rate of erythropoiesis compared with older animals. ?? 2009 American Society for Veterinary Clinical Pathology.

Harvey, J. W.; Harr, K. E.; Murphy, D.; Walsh, M. T.; Nolan, E. C.; Bonde, R. K.; Pate, M. G.; Deutsch, C. J.; Edwards, H. H.; Clapp, W. L.



Toxic effects of dietary methylmercury on immune function and hematology in American kestrels (Falco sparverius)  

USGS Publications Warehouse

Fifty-nine adult male American kestrels (Falco sparverius) were assigned to one of three diet formulations including 0 (control), 0.6, and 3.9 ?g/g (dry wt) methylmercury (MeHg). Kestrels received their diets daily for 13 weeks to assess the effects of dietary MeHg on immunocompetence. Immunotoxic endpoints included assessment of cell-mediated immunity (CMI) using the phytohemagglutinin (PHA) skin-swelling assay and primary and secondary antibody-mediated immune responses (IR) via the sheep red blood cell (SRBC) hemagglutination assay. Select hematology and histology parameters were evaluated to corroborate the results of functional assays and to assess immunosuppression of T and B cell-dependent components in spleen tissue. Kestrels in the 0.6 and 3.9 ?g/g MeHg groups exhibited suppression of CMI, including lower PHA stimulation indexes (p = 0.019) and a 42 to 45% depletion of T cell-dependent splenic lymphoid tissue (p = 0.006). Kestrels in the 0.6 ?g/g group exhibited suppression of the primary IR to SRBCs (p = 0.014). MeHg did not have a noticeable effect on the secondary IR (p = 0.166). Elevation of absolute heterophil counts (p p p = 0.003) was apparent in the 3.9 ?g/g group at week 12. Heterophilia, or the excess of heterophils in peripheral blood above normal ranges, was apparent in seven of 17 (41%) kestrels in the 3.9 ?g/g group and was indicative of an acute inflammatory response or physiological stress. This study revealed that adult kestrels were more sensitive to immunotoxic effects of MeHg at environmentally relevant dietary concentrations than they were to reproductive effects as previously reported.

Fallacara, Dawn M.; Halbrook, Richard S.; French, John B.



CD31/PECAM-1 is a ligand for alpha v beta 3 integrin involved in adhesion of leukocytes to endothelium  

PubMed Central

To protect the body efficiently from infectious organisms, leukocytes circulate as nonadherent cells in the blood and lymph, and migrate as adherent cells into tissues. Circulating leukocytes in the blood have first to adhere to and then to cross the endothelial lining. CD31/PECAM- 1 is an adhesion molecule expressed by vascular endothelial cells, platelets, monocytes, neutrophils, and naive T lymphocytes. It is a transmembrane glycoprotein of the immunoglobulin gene superfamily (IgSF), with six Ig-like homology units mediating leukocyte-endothelial interactions. The adhesive interactions mediated by CD31 are complex and include homophilic (CD31-CD31) or heterophilic (CD31-X) contacts. Soluble, recombinant forms of CD31 allowed us to study the heterophilic interactions in leukocyte adhesion assays. We show that the adhesion molecule alpha v beta 3 integrin is a ligand for CD31. The leukocytes revealed adhesion mediated by the second Ig-like domain of CD31, and this binding was inhibited by alpha v beta 3 integrin-specific antibodies. Moreover alpha v beta 3 was precipitated by recombinant CD31 from cell lysates. These data establish a third IgSF-integrin pair of adhesion molecules, CD31-alpha v beta 3 in addition to VCAM-1, MadCAM-1/alpha 4 integrins, and ICAM/beta 2 integrins, which are major components mediating leukocyte-endothelial adhesion. Identification of a further versatile adhesion pair broadens our current understanding of leukocyte-endothelial interactions and may provide the basis for the treatment of inflammatory disorders and metastasis formation.



Evaluation of a Multiplexed Bead Assay for Assessment of Epstein-Barr Virus Immunologic Status  

PubMed Central

Currently, serological assays using either indirect immunofluorescence assay or enzyme-linked immunosorbent assay (ELISA) are performed to evaluate the status of Epstein-Barr virus (EBV) infection in humans. Although these methods are reliable, they are limited to testing an antibody response to a single viral antigen per reaction, thus necessitating a panel of assays to complete the evaluation. In contrast, a new bead-based method (BioPlex 2200; Bio-Rad Laboratories, Hercules, Calif.) can analyze the humoral response to multiple antigens in a single tube. This approach potentially reduces overall cost, turnaround time, and sample volume. The aim of this study was to evaluate the multiplexed EBV serologic assays performed on the BioPlex 2200 platform compared to results of conventional heterophile and ELISA-based assays. A total of 167 nonconsecutive, stored serum samples from adult and pediatric patients submitted for EBV serologic studies were used in the evaluation. Concordance between results generated by the BioPlex 2200 system and conventional assays was calculated. The anti-EA-D assay had the lowest concordance at 91%. The BioPlex 2200 system showed 97% agreement with conventional heterophile and anti-nuclear antigen assays and 92% agreement with the anti-VCA IgG and immunoglobulin M assays. Agreement between the BioPlex 2200 system and conventional testing was 92% with respect to categorization of acute versus nonacute EBV disease. The correlation between these two systems with regard to assignment into one of four categories of EBV status was also good (82%). In summary, there is excellent correlation between contemporary EBV serologic testing and the BioPlex 2200 system.

Klutts, J. S.; Liao, R. S.; Dunne, W. M.; Gronowski, A. M.



Changes in selected biochemical indices, leukocyte profile, and pterins as biomarkers of immune system activity due to antipecking measures in pheasants.  


The physiological changes in response to beak trimming and spectacle usage as antipecking measures were monitored in 10-mo-old common pheasants (Phasianus colchicus). Short-term analysis conducted before the beginning of the laying period showed immediate increases of plasma corticosterone (P < 0.05) and lactate dehydrogenase (P < 0.001) concentrations and decrease of plasma triglycerides (P < 0.01) levels in response to both beak trimming and the application of spectacles. Beak-trimmed pheasants exhibited higher plasma corticosterone concentrations than pheasants fitted with spectacles (P < 0.001). To assess long-term changes, blood samples for biochemical (neopterin and biopterin determination) and hematological (leukocyte profile determination) examinations were taken from beak-trimmed, spectacles-fitted, and control pheasant hens housed in cages during their laying period. At the end of the laying period, hens fitted with spectacles exhibited lower concentrations of plasma neopterin (P = 0.005) and biopterin (P = 0.005) than beak-trimmed pheasant hens. Our findings suggest that the immune system was suppressed in spectacles-fitted pheasant hens as a result of chronic stress, as also indicated by the higher heterophil-to-lymphocyte ratio (P = 0.001) compared with beak-trimmed hens. Our study found a negative correlation (r = -0.31, P = 0.019) between the heterophil-to-lymphocyte ratio and plasma neopterin concentration. This study demonstrated that both beak trimming and use of spectacles are not only stressful procedures for pheasants, but long-term effects may also include a negative impact on the immune system. PMID:23776255

Voslarova, E; Bedanova, I; Pistekova, V; Marsalek, P; Chloupek, J



Feeding the BT cationic peptides to chickens at hatch reduces cecal colonization by Salmonella enterica serovar Enteritidis and primes innate immune cell functional activity.  


The BT/TAMUS 2032 (BT) cationic peptides are a group of related cationic peptides produced by a Gram-positive soil bacterium, Brevibacillus texasporus. Cationic amphiphilic peptides produced by host cells have been found to stimulate or prime the innate immune responses in mammals, but little information is available on the effects of bacterial-produced peptides on host immunity. We have previously shown that BT, provided as a feed additive for 4 days after hatch, significantly induced protection against extraintestinal colonization by Salmonella enterica serovar Enteritidis. We also found that feeding BT significantly upregulated the functional efficiency of heterophils, the avian equivalent to mammalian neutrophils. The objective of the present study was to further evaluate the effect of BT as a nonantibiotic, antibacterial compound and a stimulator of the innate immune response of young chickens. BT, provided as a feed additive at three different concentrations (12, 24, or 48 ppm) for 4 days after hatch, significantly increased protection against Salmonella enterica serovar Enteritidis cecal colonization in a concentration-dependent manner. We also confirmed our previous results that the functional activities of heterophils from chickens fed the BT rations were significantly upregulated. In addition, we also found that the functional activities of peripheral blood monocytes were significantly increased in a concentration-dependent manner when compared with monocytes isolated from chickens fed a control diet. This is the first report of bacterial cationic peptides providing protection against Salmonella cecal colonization. The significance of these data is that the orally delivered cationic peptides stimulate the innate response during the first week after hatch, normally a time of immunologic inefficiency and increased susceptibility to bacterial infections. We speculate that BT given as a feed additive during the first week after hatch could provide increased protection against a variety of bacterial pathogens because of the nonspecific nature of the innate response. PMID:19735207

Kogut, Michael H; He, Haiqi; Genovese, Kenneth J; Jiang, Yiwei W



Effects of long-term heat stress in an experimental model of avian necrotic enteritis.  


Stressful conditions are predisposing factors for disease development. Heat stress is one of the most important stressors in poultry production. The reemergence of some previously controlled diseases [e.g., avian necrotic enteritis (NE)] has been extensively reported. The combination of bacterial infection and certain environmental factors have been reported to trigger the disease. The aim of this study was to analyze the effects of long-term heat stress (35 ± 1°C) on the development of NE in broiler chickens. For this purpose, 60 male broiler chickens were divided into the following 6 groups: control group (C), heat stressed control group (C/HS35), thioglycolate group (T), thioglycolate heat-stressed group (T/HS35), infected group (I), and infected heat-stressed group (I/HS35). The poultry of groups I and I/HS35 were experimentally infected with Clostridium perfringens via their feed from 15 to 21 d of life. Heat stress (35 ± 1°C) was constantly applied to the birds of the stressed groups from 14 to 21 d of life. The infected and heat-stressed broiler chickens presented a trend toward a decrease in gross lesion scores and significantly lower microscopic scores of necrosis in the duodenum and jejunum (P < 0.05), lower fusion of villi in the duodenum (P < 0.05), and lower congestion scores in the jejunum and ileum (P < 0.05) in relation to infected and non-heat-stressed chickens. Broilers of I/HS35 group also exhibited small number of heterophils in the duodenum and jejunum compared with those of the I group (P < 0.05). Furthermore, the duodenum and jejunum of infected and heat-stressed broilers showed lower number of clostridia on the intestinal mucosa (P < 0.05). Data were discussed in light of a heat stress induced reduction on intestinal inflammation via a decrease in heterophil migration to the intestinal mucosa, which in turn might have reduced tissue damage during inflammation, hence preventing the development of a more severe form of NE. PMID:24879684

Calefi, Atilio Sersun; Honda, Bruno Takashi Bueno; Costola-de-Souza, Carolina; de Siqueira, Adriana; Namazu, Lilian Bernadete; Quinteiro-Filho, Wanderley Moreno; Fonseca, Juliana Garcia da Silva; Aloia, Thiago Pinheiro Arrais; Piantino-Ferreira, Antonio José; Palermo-Neto, João



Bilateral acute pyogenic conjunctivitis with iritis induced by unilateral topical application of bacterial peptidoglycan muramyl dipeptide in adult rabbits.  


The factors responsible for the conjunctivitis and iritis associated with acute ocular infection and post enteric inflammatory disease are not fully known. The pro-inflammatory activity of unilateral topical application of muramyl dipeptide (MDP; the smallest bio-active Gram-positive and Gram-negative bacterial cell wall component) was investigated in adult rabbits. The resultant bilateral conjunctivitis/iritis and pyogenic responses were characterized. Bilateral symptoms were graded by slit lamp examinations; tear fluid, Schirmer tests (tear production), blood and aqueous humor (AH) samples were obtained from MDP-treated and untreated rabbits. MDP concentration, gamma-glutamyltranspeptidase activity (GGT; key enzyme in glutathione recapture, xenobiotic detoxification, eicosanoid synthesis and neutrophil function), protein concentration, and tear cell density, cytology, and immunofluorescent antibody reactivity to GGT and calreticulin (CRT; MDP-binding protein) were determined. MDP was cleared from ipsilateral tears and serum by 6 h, but was undetected in mock-treated contralateral tears. Bilateral signs of acute transient pyogenic conjunctivitis, characterized by tearing, lid edema, conjunctival hyperemia, chemosis and leukocytic infiltrate with iritis (erythema and aqueous flare) were detected. Milder symptoms occurred in the mock-treated contralateral eyes. Bilateral symptoms, tear production, tear protein, GGT activity, and mucopurulent discharge (containing up to 2.5-5.0 × 10(6) cells/mL) were elevated 4-8 h post MDP and resolved to near pre-treatment levels by 24 h. Tear GGT activity and protein levels were higher in MDP-treated and mock-treated contralateral eyes than in eyes of untreated adult rabbits (p's < 0.001). Elevated tear GGT activity was associated with histopathology and increased vascular and epithelial permeability to serum protein, GGT-positive epithelia cells, macrophages and heterophils. Repeat MDP applications induced recurrent induction and resolution patterns of bilateral conjunctivitis/iritis and tear GGT activity, but ipsilateral GGT responses were lower. The results suggest unilateral topical MDP application to adult rabbit eyes induces a bilateral acute pyogenic conjunctivitis/iritis (PCI) characterized by increased vascular and epithelial permeability similar to acute bacterial conjunctivitis in man. The detection of CRT/GGT positive heterophils in tears suggests efferocytosis (phagocytosis of dead/dying cells). Tear GGT activity may be a useful means to quantify MDP-induced toxicity and extraocular inflammation. PMID:24135297

Langford, Marlyn P; Foreman, Bridgett D; Srur, Lana; Ganley, James P; Redens, Thomas B



Understanding stress-induced immunosuppression: exploration of cytokine and chemokine gene profiles in chicken peripheral leukocytes.  


At present, the poultry meat and egg industry has gained a lot of ground, being viewed as a provider of a healthy alternative to red meat and other protein sources. If this trend is to be maintained, solutions must be found to improve resistance of chickens to disease, which often is weakened by stressful conditions. In poultry, stress-induced immunosuppression is manifested by failures in vaccination and increased morbidity and mortality of flocks. Currently, several modern cellular and molecular approaches are being used to explore the status of the immune system during stress and disease. It is likely that these new techniques will lead to the development of new strategies for preventing and controlling immunosuppression in poultry. Using quantitative reverse transcription-PCR assays, a broad spectrum of cytokine, chemokine, and their receptor genes can be quantified in birds and then be used as markers to assess the effects of stress on the immune system. Currently, we are investigating immune and endocrine interactions in the chicken, in particular the cells and molecules that are known to be involved in such interactions in mammals. We have evaluated the effects of corticosterone administration in drinking water on peripheral lymphocyte and heterophil cytokine and chemokine gene profiles. In particular, there seems to be effects on cytokine and chemokine mRNA expression levels in both lymphocytes and heterophils, especially expression of the proinflammatory cytokines interleukin (IL)-1beta, IL-6, and IL-18 and chemokines C-C motif, ligand 1 inflammatory (CCLi1); C-C motif, ligand 2 inflammatory (CCLi2); C-C motif, ligand 5 (CCL5); C-C motif, ligand 16 (CCL16); C-X-C motif ligand 1 inflammatory (CXCLi1); and C-X-C motif ligand 2 inflammatory (CXCLi2), which are initially upregulated and are potentially involved in modulating the adaptive immune response. A chronic treatment with corticosterone downregulates proinflammatory cytokines and chemokines, suggesting that the delayed effects of chronic stress can suppress the immune response. Messenger RNA expression levels of transforming growth factor-beta4 (TGF-beta4) are also upregulated in cortisosterone-treated birds. It appears that the balance between T-helper (Th) 1 and Th2/T regulatory cytokine production is altered in conditions associated with significant changes in plasma corticosterone concentration. Experiments are underway to decipher the cytokine and chemokine responses to vaccination and bacterial challenge on the background of stress-induced immunosuppression. PMID:20308420

Shini, S; Huff, G R; Shini, A; Kaiser, P



Ecological immunology in a fluctuating environment: an integrative analysis of tree swallow nestling immune defense  

PubMed Central

Evolutionary ecologists have long been interested by the link between different immune defenses and fitness. Given the importance of a proper immune defense for survival, it is important to understand how its numerous components are affected by environmental heterogeneity. Previous studies targeting this question have rarely considered more than two immune markers. In this study, we measured seven immune markers (response to phytohemagglutinin (PHA), hemolysis capacity, hemagglutination capacity, plasma bactericidal capacity, percentage of lymphocytes, percentage of heterophils, and percentage of eosinophils) in tree swallow (Tachycineta bicolor) nestlings raised in two types of agro-ecosystems of contrasted quality and over 2 years. First, we assessed the effect of environmental heterogeneity (spatial and temporal) on the strength and direction of correlations between immune measures. Second, we investigated the effect of an immune score integrating information from several immune markers on individual performance (including growth, mass at fledging and parasite burden). Both a multivariate and a pair-wise approach showed variation in relationships between immune measures across years and habitats. We also found a weak association between the integrated score of nestling immune function and individual performance, but only under certain environmental conditions. We conclude that the ecological context can strongly affect the interpretation of immune defenses in the wild. Given that spatiotemporal variations are likely to affect individual immune defenses, great caution should be used when generalizing conclusions to other study systems.

Pigeon, Gabriel; Belisle, Marc; Garant, Dany; Cohen, Alan A; Pelletier, Fanie



Serological diagnosis of Epstein-Barr virus infection: Problems and solutions  

PubMed Central

Serological tests for antibodies specific for Epstein-Barr virus (EBV) antigens are frequently used to define infection status and for the differential diagnosis of other pathogens responsible for mononucleosis syndrome. Using only three parameters [viral capsid antigen (VCA) IgG, VCA IgM and EBV nuclear antigen (EBNA)-1 IgG],it is normally possible to distinguish acute from past infection: the presence of VCA IgM and VCA IgG without EBNA-1 IgG indicates acute infection, whereas the presence of VCA IgG and EBNA-1 IgG without VCA IgM is typical of past infection. However, serological findings may sometimes be difficult to interpret as VCA IgG can be present without VCA IgM or EBNA-1 IgG in cases of acute or past infection, or all the three parameters may be detected simultaneously in the case of recent infection or during the course of reactivation. A profile of isolated EBNA-1 IgG may also create some doubts. In order to interpret these patterns correctly, it is necessary to determine IgG avidity, identify anti-EBV IgG and IgM antibodies by immunoblotting, and look for heterophile antibodies, anti-EA (D) antibodies or viral genome using molecular biology methods. These tests make it possible to define the status of the infection and solve any problems that may arise in routine laboratory practice.

De Paschale, Massimo; Clerici, Pierangelo



Great tits Parus major trade health for reproduction.  


Reproduction and maintenance compete for resources within a single individual. But do individuals invest in reproduction just as much as remains after the requirements of maintenance are covered, or do they sacrifice their health for the sake of still further increase the investment in current reproduction? This question has been found hard to answer because of difficulties of demonstrating that individuals naturally make a reproductive effort of such a magnitude as to inflict health damage. In this paper we present evidence for a trade-off between reproductive effort and health state in great tits, indicated by a positive correlation between total prefledging brood weight and both intensity of Haemoproteus blood parasite infection and heterophile: lymphocyte (H:L) ratio. H:L ratios, which signal stress in birds, were high both among individuals making an intense reproductive effort and among aberrantly behaving individuals, such as females incubating in empty nests and birds which abandoned their broods after blood sampling. Experimental reduction of clutch size resulted in decreased intensity of Haemoproteus parasitemia, providing further evidence that individual great tits accept immunosuppression to increase their reproductive investment. PMID:8952087

Ots, I; Hõrak, P



From Split to Sibenik: The Tortuous Pathway in the Cholinesterase Field  

PubMed Central

The interim between the first and tenth International Cholinesterase meetings has seen remarkable advances associated with the applications of structural biology and recombinant DNA methodology to our field. The cloning of the cholinesterase genes led to the identification of a new super family of proteins, termed the ?,?–hydrolase fold; members of this family possess a four helix bundle capable of linking structural subunits to the functioning globular protein. Sequence comparisons and three dimensional structural studies revealed unexpected cousins possessing this fold that, in turn, revealed three distinct functions for the ?,?-hydrolase proteins. These encompass: (1) a capacity for hydrolytic cleavage of a great variety of substrates, (2) a heterophilic adhesion function that results in trans-synaptic associations in linked neurons, (3) a chaperone function leading to stabilization of nascent protein and its trafficking to an extracellular or secretory storage location. The analysis and modification of structure may go beyond understanding mechanism, since it may be possible to convert the cholinesterases to efficient detoxifying agents of organophosphatases assisted by added oximes. Also, the study of the relationship between the ?,?–hydrolase fold proteins and their biosynthesis may yield means by which aberrant trafficking may be corrected, enhancing expression of mutant proteins. Those engaged in cholinesterase research should take great pride in our accomplishments punctuated by the series of ten meetings. The momentum established and initial studies with related proteins all hold great promise for the future.

Taylor, Palmer



Different roles of cadherins in the assembly and structural integrity of the desmosome complex.  


Adhesion between cells is established by the formation of specialized intercellular junctional complexes, such as desmosomes. Desmosomes contain isoforms of two members of the cadherin superfamily of cell adhesion proteins, desmocollins (Dsc) and desmogleins (Dsg), but their combinatorial roles in desmosome assembly are not understood. To uncouple desmosome assembly from other cell-cell adhesion complexes, we used micro-patterned substrates of Dsc2aFc and/or Dsg2Fc and collagen IV; we show that Dsc2aFc, but not Dsg2Fc, was necessary and sufficient to recruit desmosome-specific desmoplakin into desmosome puncta and produce strong adhesive binding. Single-molecule force spectroscopy showed that monomeric Dsc2a, but not Dsg2, formed Ca(2+)-dependent homophilic bonds, and that Dsg2 formed Ca(2+)-independent heterophilic bonds with Dsc2a. A W2A mutation in Dsc2a inhibited Ca(2+)-dependent homophilic binding, similar to classical cadherins, and Dsc2aW2A, but not Dsg2W2A, was excluded from desmosomes in MDCK cells. These results indicate that Dsc2a, but not Dsg2, is required for desmosome assembly through homophilic Ca(2+)- and W2-dependent binding, and that Dsg2 might be involved later in regulating a switch to Ca(2+)-independent adhesion in mature desmosomes. PMID:24610950

Lowndes, Molly; Rakshit, Sabyasachi; Shafraz, Omer; Borghi, Nicolas; Harmon, Robert M; Green, Kathleen J; Sivasankar, Sanjeevi; Nelson, W James



Reference values for hematology, plasma biochemical analysis, plasma protein electrophoresis, and Aspergillus serology in elegant-crested tinamou (Eudromia elegans).  


Elegant-crested tinamou (Eudromia elegans) are members of the family Tinamidae. A review of postmortem records at the Saint Louis Zoo suggests that this species is highly susceptible to aspergillosis. Antemortem detection of Aspergillus infections can be aided by evaluating results of hematologic, plasma biochemical, protein electrophoretic, and Aspergillus antigen and antibody serologic testing. To establish species-specific reference intervals for these parameters in elegant-crested tinamou, blood samples were collected and analyzed from 19 healthy captive adult and subadult elegant-crested tinamou. The predominant circulating leukocyte in elegant-crested tinamou is the lymphocyte. Prealbumin electrophoresis fractions were detected in 4 of 19 birds (21%), and globulin fractions were detected in all birds. All birds had measurable titers for Aspergillus antibody and galactomannan antigen, including 3 birds with antibody levels considered to be positive. Galactomannan levels were all below the positive threshold of 1.0. Results of hematologic and plasma biochemical analyses performed on blood samples obtained from terminally ill elegant-crested tinamou with confirmed aspergillosis were compared with results from healthy individuals. Significant differences were found in the hematocrit, total white blood cell count, heterophil and monocyte counts, and albumin and globulin levels. PMID:23772450

Black, Peter A; Macek, Michael; Tieber, Anne; Weber, Martha



Clinical haematology of the great bustard (Otis tarda).  


The haematological parameters of healthy great bustards (Otis tarda L.) have been determined. The values obtained were red cell count (3.0 x 10(12) +/- 0.2 x 10(12/)1), white cell count (33.0 x 10(9) +/- 2.6 x 10(9)/1), haematocrit value (0.51 +/- 0.01 1/1), haemoglobin (13.0 +/- 0.3 g/dl), mean corpuscular volume (178.7 +/- 12.5 fl), mean cell haemoglobin concentration (25.0 +/- 0.6 g/dl), mean corpuscular haemoglobin (42.5 +/- 3.2 pg), differential white cell count: heterophils (22.5 x 10(9) +/- 0.7 x 10(9)/1), lymphocytes (6.0 x 10(9)+/-0.7 x 10(9)/1), eosinophils (2.7 x 10(9) +/- 0.3 x 10(9)/1) and monocytes (1.8 x 10(9)+/-0.2 x 10(9)/1). PMID:18680064

Jimenez, A; Barrera, R; Sanchez, J; Cuenca, R; Rodriguez, J; Andres, S; Mane, M C



Impairment of growth and immune function of avocet chicks from sites with elevated selenium, arsenic, and boron.  


Avocets (Recurvirostra americana) hatched from eggs collected from the south Central Valley of California (USA) were studied to determine the impact of elevated concentrations of selenium, arsenic, and boron on the immune system and growth to maturity. Corcoran ponds were the reference site with low selenium (1.2 ppb) and arsenic (29 ppb) (boron not measured). Westfarmers Pond had elevated concentrations of selenium (319 ppb), arsenic (127 ppb), and boron (109 ppm). Pryse ponds also had elevated selenium, arsenic, and boron concentrations (13.9 ppb, 1,100 ppb, and 29.4 ppm, respectively). Size at hatch was significantly reduced (P < or = 0.05) in birds from Westfarmers and Pryse ponds. The growth rate was faster, but mean adult size was reduced in birds from Pryse ponds. Avocet chicks from Pryse and Westfarmers ponds exposed solely through in ovo transfer of these elements had significantly increased heterophil:lymphocyte ratios. The phagocytic activity of macrophages also was significantly reduced in these birds, and Pryse Pond birds had an increased proliferative ability of lymphocytes in the presence of concanavalin A, a T-cell mitogen. Avocet chicks (< or = 5 wk old) were captured from the various ponds and the same morphometric and immune function measurements made. The birds that were most severely impacted by exposure to these compounds were those that were collected from Pryse ponds. PMID:8028107

Fairbrother, A; Fix, M; O'Hara, T; Ribic, C A



Variation in stress and innate immunity in the tree lizard (Urosaurus ornatus) across an urban-rural gradient  

PubMed Central

The urban environment presents new and different challenges to wildlife, but also potential opportunities depending on the species. As urban encroachment onto native habitats continues, understanding the impact of this expansion on native species is vital to conservation. A key physiological indicator of environmental disturbance is the vertebrate stress response, involving increases in circulating glucocorticoids (i.e., corticosterone), which exert influence on numerous physiological parameters including energy storage, reproduction, and immunity. We examined how urbanization in Phoenix, Arizona influences corticosterone levels, blood parasitism, and innate immunity in populations of tree lizards (Urosaurus ornatus) to determine whether urbanization may be detrimental or beneficial to this species. Both baseline and stress-induced corticosterone concentrations were significantly lower in urban lizards relative to the rural ones, however, the magnitude of the increase in corticosterone with stress did not differ across populations. Urban lizards also had a lower ratio of heterophils to lymphocytes, but elevated overall leukocyte count, as compared to lizards from the natural site. Urban and rural lizards did not differ in their prevalence of the blood parasite, Plasmodium mexicanum. Taken together, these results suggest that urban tree lizards may have suppressed overall corticosterone concentrations possibly from down-regulation as a result of frequent exposure to stressors, or increased access to urban resources. Also, urban lizards may have bolstered immunocompetence possibly from increased immune challenges, such as wounding, in the urban environment, or from greater energetic reserves being available as a result of access to urban resources.

French, Susannah S.; Fokidis, H. Bobby; Moore, Michael C.



A new report on the occurrence of clinico-histopathological diagnostic criteria for Spinodiplotriaena.  


Parasitic diseases of wild birds follows its companion volume, infectious diseases of wild birds, both of which complement the third edition of infectious diseases of wild mammals and second edition of parasitic diseases of wild mammals. In October 2012, a mature black male mynah with lack of balance and ataxia was brought into the birds department of educational hospital in veterinary medicine faculty of Tehran University. Because of no cure in clinical signs after several days of empirical therapy euthanasia and clinical pathology studies were performed. There was no microscopic pathological lesions in brain, bone marrow, heart, lung, liver and the microscopic results were as following; bone marrow: hemophagocytosis reactive macrophages, liver: infiltration of inflammatory cells (hepatitis) and hemosiderin-laden macrophages. Lung: hemosiderin-laden macrophages and spleen findings: infiltration of inflammatory cells, heterophils with phagocytic bacteria. In abdominal cavity two male and one female parasite was detect. Three parasites were cleared with lactophenol and investigated under the light microscopy and important morphological parts of them measured. Also these parasites in serum containing send to the department of parasitology. Based on all collecting data, Spinodiplotriaena (three parasites each by the length of 2/5, 1/7 and 1/5 cm were found) were diagnosed. Report of this case in important because of first report of this infection in a black mynah in Iran. PMID:25035597

Javanbakht, Javad; Hosseini, Ehsan; Karimi, Shahram; Hassan, Mehdi Aghamohammad; Sabbagh, Atefeh; Mousavi, Shadi; Fattahi, Roohollah; Khaki, Fariba; Shafiee, Radmehr



Mucosal injury and. gamma. -irradiation produce persistent gastric ulcers in the rabbit. Evaluation of antiulcer drug binding to experimental ulcer sites  

SciTech Connect

A method producing persistent gastric ulcers in the rhesus monkey by combined mucosal injury and {gamma}-irradiation was modified and evaluated in the rabbit. {gamma}-Irradiation (800-1000 cGy) immediately after removal of 2-mm-diameter sections of antral mucosa resulted in ulcer craters 5-7 days later. Ulcer sites were characterized by loss of the mucosa, muscularis mucosa, and much of the submucosa. The exposed submucosa was coated with fibrin and necrotic debris infiltrated with heterophils, the rabbit equivalent of neutrophils. These ulcers strongly resemble human chronic gastric ulcers. Binding of Carafate (sucralfate; Marion Laboratories, Inc., Kansas City, MO) and Maalox (magnesia-alumina oral suspension; Wm. H. Rorer, Inc., Ft. Washington, PA) to ulcer and nearby nonulcer sites in the antrum was assessed 1 hour after drug dosing. Drug binding was determined by aluminum quantitation of stomach wall punch biopsies at necropsy. Both drugs significantly increased aluminum bound to the stomach wall compared with vehicle treatment. Significantly more antiulcer drug was bound to ulcer sites than to nearby nonulcer sites only after sucralfate administration. This model of persistent gastric ulcer should be useful to further study gastric ulcer pathogenesis and treatment.

Yokel, R.A.; Dickey, K.M. (Univ. of Kentucky, Lexington (USA))



Haemopoiesis in the head kidney of tilapia, Oreochromis niloticus (Teleostei: Cichlidae): a morphological (optical and ultrastructural) study  

PubMed Central

The present work focused on the histological and ultrastructural studies on haemopoiesis in the kidney of tilapia, Oreochromis niloticus. Haemopoietic tissue was found mainly in the head kidney and a small amount occurred in the mesonephros. The haemopoiesis of tilapia had the following series: erythropoiesis, granulopoiesis, thrombopoiesis, monopoiesis and lymphoplasmopoiesis. Erythropoiesis includes proerythroblasts, basophilic erythroblasts, polychromatic erythroblasts, acidophilic erythroblasts and young and mature erythrocytes. The proerythroblasts were the largest cells in the erythropoietic series. During the maturation process both the nuclear and cellular size decreased gradually due to the chromatin condensation and the progressive substitution of cytoplasmic matrix with a large amount of haemoglobin. Granulopoietic series consisted of cells with variable shape and size at different stages of maturity from myeloblasts to mature granulocytes. The promyelocytes were the largest cells in the series and were characterised by the appearance of primary (azoruphilic) granules. The maturation process involved the appearance of specific granules in the heterophilic, eosinophilic and basophilic series. It is important to mention that eosinophilic granulocytes were the dominant granulopoietic series in the haemopoietic tissue (Ht) of tilapia. Lymphopoietic series consisted of lymphoblasts, large lymphocytes, small lymphocytes and active and inactive plasma cells. Thrombopoietic series consisted of thromboblasts, prothromboblasts and thrombocytes. Thrombocytes of tilapia were nucleated and possessed a spindle shape. Melanomacrophage centres were dominant among the Ht of the head kidney. Also, monocytes were detected and shown to be large cells with an indented nucleus and cytoplasm containing numerous vesicles of different sizes and a few lysosomes.

Abdel-Aziz, El-Saydah H.; Abdu, Suzan B. S.; Fouad, Huda F.



Thyrotoxicosis presenting as hypogonadism: a case of central hyperthyroidism.  


Herein, we present a case of central thyrotoxicosis with well-documented serial therapeutic interventions. Thyroid-stimulating hormone (TSH)-secreting pituitary tumors represent a rare cause of hyperthyroidism. It is being diagnosed more frequently with the third-generation TSH assay. Many conditions can produce normal or elevated TSH levels in combination with elevated thyroid hormone levels. The differential diagnosis includes resistance to thyroid hormone (RTH, Refetoff's syndrome), assay interference from anti-T4/T3 and heterophile antibodies, elevated or altered binding proteins, drugs affecting peripheral metabolism, and noncompliance with thyroid replacement therapy. In contrast to RTH, our patient presented had high alpha-subunit-to-TSH molar ratio, failed TSH response to thyrotropin-releasing hormone stimulation, and a large pituitary mass. Normal or high TSH in the presence of elevated T4 or T3 is a fairly common clinical scenario with many etiologic possibilities. This TSH-producing adenoma represents an unusual initial clinical presentation, as hypogonadism appeared before features of thyrotoxicosis were appreciated. This case represents the most modern therapeutic approach to the management of this rare disease. Our patient has done well on octreotide with control of thyrotoxicosis and an additional 30% shrinkage of his tumor mass. PMID:15545848

Childress, R Dale; Qureshi, M Nauman; Kasparova, Meri; Oktaei, Hooman; Williams-Cleaves, Beverly; Solomon, Solomon S



Growth and metabolic characterization of Macrorhabdus ornithogaster.  


Macrorhabdus ornithogaster (M. ornithogaster) is an anamorphic ascomycetous yeast found only in the stomach of birds. Infection is often benign but has also been associated with disease in some species of birds under some circumstances. In vitro efforts to grow M. ornithogaster have been largely unsuccessful. In this report, multiple liquid and solid media of varying pH, sugar concentration, and fetal bovine serum (FBS) concentrations, incubated at various temperatures in room air or microaerophilic conditions, were examined for their ability to support the growth of M. ornithogaster, obtained from a budgerigar (Melopsittacus undulatus). Optimum growth conditions were found to be Basal Medium Eagle's, pH 3 to 4, containing 20% FBS, and 5% glucose or sucrose under microaerophilic conditions at 42 degrees C. Using these conditions, M. ornithogaster was repeatedly passaged without loss of viability. Polyclonal isolates of M. ornithogaster consistently assimilated glucose, sucrose, and trehalose. M. ornithogaster did not grow with prolonged exposure to atmospheric oxygen, but growth in microaerophilic conditions was moderately enhanced by preincubation with atmospheric oxygen for 24 hours. An isolate of M. ornithogaster was found to be infective to day-old chickens, reduce their rate of weight gain, and induce a mild to moderate heterophilic inflammation of the isthmus. M. ornithogaster was reisolated from the chicks 7 days after infection, fulfilling Koch's postulates. A 761-bp sequence of 18S rDNA from this isolate was compared to the originally reported M. ornithogaster sequence and was found to be 97% identical. PMID:17459854

Hannafusa, Yasuko; Bradley, Allison; Tomaszewski, Elizabeth E; Libal, Melissa C; Phalen, David N



Assessment of multiple cardiocentesis in ball pythons (Python regius).  


This study evaluated the gross and microscopic effects of serial blood collection from six ball pythons (Python regius) by using cardiocentesis. We collected 39 blood samples from each snake over a 120-day period. Cardiocentesis was performed on manually restrained snakes, with each sample requiring approximately 15 sec to collect. No clinically apparent complications were noted in any of the snakes after the cardiocentesis procedures, and all snakes survived until they were euthanized 73 days after the last blood sample. Minimal gross lesions were noted at necropsy; faint brown pigmentation of the pericardium was present in five of six snakes, and three snakes had approximately 0.5 ml dark pigmented fluid in the pericardial space. One snake had a small, organized hematoma in the pericardial space. Microscopic findings were limited to moderate and regularly arranged collagen fibrosis and focal thickening of the epicardium. The pericardial sac in all snakes had a mild infiltrate of hemosiderin-laden macrophages and small numbers of heterophils. The results suggest that serial cardiocentesis is well tolerated in ball pythons. PMID:15636554

Isaza, Ramiro; Andrews, Gordon; Coke, Rob; Hunter, Robert



Retrobulbar adenocarcinoma in an Amazon parrot (Amazona autumnalis).  


Retrobulbar neoplasms are not common in mammals and are even more infrequently seen in nonmammalian species. The current report describes a retrobulbar mass creating exophthalmia and neurologic signs in a red-lored Amazon parrot (Amazona autumnalis). A 27-year-old female parrot presented for a 3-day history of anorexia and a 2-week history of periocular soft tissue swelling and exophthalmia of the right eye. Physical examination revealed 9% dehydration and right eye exophthalmia with inability to retropulse the globe. A fine-needle aspirate was performed, and cytologic evaluation revealed necrotic debris with scattered clusters of epithelial cells, moderate numbers of macrophages, and few heterophils. Given the possibility of neoplasia and paucity of treatment options, the owners elected euthanasia and submitted the body for necropsy. A large, fluctuant, friable, red, retrobulbar mass with multiple areas of hemorrhage, on cut surface, was noted at necropsy. Histologically, the mass was composed of neoplastic, cuboidal to columnar epithelial cells, forming rosette-like glandular structures, admixed with abundant necrotic debris. The neoplastic cells were strongly positive for cytokeratin (AE1/AE3) by immunohistochemistry. Based on histopathology and immunohistochemistry, the mass was diagnosed as an adenocarcinoma. PMID:23404475

Watson, Victoria E; Murdock, Jessica H; Cazzini, Paola; Schnellbacher, Rodney; Divers, Stephen J; Sakamoto, Kaori



Clinical and pathological findings of concurrent poxvirus lesions and aspergillosis infection in canaries  

PubMed Central

Objective To investigate clinical, pathological and mycological findings in canaries, in which pox lesions and Aspergillus fumigatus (A. fumigatus) infection were observed simultaneously. Methods This study was performed on a breeding colony (about 100 canaries) affected by fatal wasting disease. Necropsy was undertaken on 10 severely affected canaries, and gross lesions were recorded. Samples from internal organs displaying lesions were obtained for histopathological evaluation. Tracheal swap samples of internal organs of the all infected animals with lesions at necropsy were cultured in Sabouraud Dextrose Agar for mycological examination. Results At necropsy, caseous foci were determined in the lungs, on the air sacs, liver, spleen, heart. Swelling of the eyelids, diffuse hemorrhages in the subcutaneous tissue with small papular lesions of the skin were other typical necropsy findings. Histopathologically, pathognomonic eosinophilic intracytoplasmic inclusion bodies, which called Bollinger bodies, in both skin cells and vacuolated air way epithelial cells confirmed canary pox infection. Moreover, histopathological examination of the white-yellowish caseous foci revealed necrotic granulomatous reaction consisting of macrophages, heterophil leukocytes and giant cells encapsulated with a fibrous tissue. After the culture of the tissue samples, the formation of bluish green colonies confirmed A. fumigatus infection. Conclusions Canary pox has been known as the disease that can result in high losses in a short time, as a re-emerging disease that has not been present during recent years in canary flocks in Iran. So, the current paper provides useful information to prevent misdiagnosed of canary pox disease which can cause secondary mycotic infection.

Reza, Kheirandish; Nasrin, Askari; Mahmoud, Salehi



Low temperatures reduce skin healing in the Jacaré do Pantanal (Caiman yacare, Daudin 1802).  


Studies of skin wound healing in crocodilians are necessary given the frequent occurrence of cannibalism in intensive farming systems. Air temperature affects tissue recovery because crocodilians are ectothermic. Therefore, the kinetics of skin wound healing in Caiman yacare were examined at temperatures of 33°C and 23°C. Sixteen caiman were selected and divided into two groups of eight maintained at 23°C or 33°C. The studied individuals' scars were photographed after 1, 2, 3, 7, 15 and 30 days of the experimental conditions, and samples were collected for histological processing after 3, 7, 15 and 30 days. Macroscopically, the blood clot (heterophilic granuloma) noticeably remained in place covering the wound longer for the caiman kept at 23°C. Microscopically, the temperature of 23°C slowed epidermal migration and skin repair. Comparatively, new blood vessels, labeled using von Willebrand factor (vWF) antibody staining, were more frequently found in the scars of the 33°C group. The collagen fibers in the dermis were denser in the 33°C treatment. Considering the delayed healing at 23°C, producers are recommended to keep wounded animals at 33°C, especially when tanks are cold, to enable rapid wound closure and better repair of collagen fibers because such lesions tend to compromise the use of their skin as leather. PMID:24244853

Pressinotti, Leandro Nogueira; Borges, Ricardo Moraes; Alves De Lima, Angela Paula; Aleixo, Victor Manuel; Iunes, Renata Stecca; Borges, João Carlos Shimada; Cogliati, Bruno; Cunha Da Silva, José Roberto Machado



Cryptosporidium baileyi--infection in Red-breasted Merganser (Mergus serrator) ducklings from a zoological garden.  


Infection of the Bursa of Fabricius with Cryptosporidium baileyi was diagnosed in a group of hand reared Red-breasted Merganser (Mergus serrator) ducklings during one breeding season in a German zoological garden.The birds had died in an emaciated and anaemic state after problems with spontaneous feeding. The bursae were infected with moderate to high numbers of cryptosporidia, which were associated with hyperplasia, degeneration and sloughing of the affected epithelial cells and mild heterophilic bursitis, lesions typically seen in bursal cryptosporidiosis in other avian species. In addition, lymphatic tissue was nearly absent, which was probably caused by chronic stress and malnutrition related to the rearing of these highly stress-sensitive birds in an artificial environment. Companion ducklings from stress-resistant, spontaneously feeding species from the zoological collection were used to calm the Mergansers, but may have introduced the cryptosporidia into the rearing boxes. Another possible source for the introduction of C. baileyi were adult mallards (Anas platyrhynchos), which were used to increase the hatching rate by a phase of natural breeding in the middle third of the incubation period of the Merganser eggs. PMID:23045806

Schulze, Christoph; Kämmerling, Jens; Kutzer, Peter; Engelhardt, Andreas; Richter, Barbara




PubMed Central

Scholarly tailgating and developmental relationships can soar professional identity, advance one’s scholarly career and foster cultural diversity in healthcare. This paper provides the definition and basis of scholarly tailgating for scholars who wish to propel their careers and help enable diverse cultural approaches. Scholarly tailgating is for those who wish to succeed and capitalize on the culturally diverse expertise and reciprocal network relationships from others in their web of influence. The concept of scholarly tailgating explains the use of homophilic and diverse, multidisciplinary network relationships to foster an intellectual community in healthcare in general and to benefit healthcare consumers and all being served. Scholars can benefit from standing on the shoulders of giants and allowing the aerodynamic wind to propel them to soaring career heights. Prudent scholars, apart from utilizing homophilic relationships and balancing these relationships with diverse heterophilic relationships, are able to lead in the professional academy, and become innovators who practice with cultural relevance. Multi-ethnic, diverse scholars working collaboratively can assist in igniting conversation and actions that reduce disparities, which are indigenous to those who are most vulnerable. Purposeful strategic planning and development of diverse networks that support scholarly advancement can best achieve these career accomplishments.

Adegbola, Maxine



Integrated immunogenomics in the chicken: deciphering the immune response to identify disease resistance genes.  


Resistance to infection takes place at many levels, and involves both non-specific and specific immune mechanisms. The chicken has a different repertoire of immune genes, molecules, cells and organs compared to mammals. To understand the role of any disease resistance gene(s), it is therefore important to understand these different repertoires, and the bird's response to a particular pathogen. Our studies focus on the innate immune response, as responses of macrophages from inbred lines of chickens, and heterophils from commercial birds, correlate with resistance or susceptibility to Salmonella infection with a variety of Salmonella serovars and infection models. To map disease resistance genes, we are using a combination of expression quantitative trait loci (eQTLs) from microarray studies, allied with whole genome SNP arrays (WGA) and a candidate gene approach. There are over 500 human genes with the Gene Ontology term "innate immunity". We have identified over 400 of these genes in the chicken genome, and are actively identifying informative SNPs in them. The segregation of 6000 WGA SNPs across all of our inbred lines was also assessed, which should yield approximately 900 informative SNPs for a cross between any two lines. The initial focus of these studies is on mapping resistance genes in our inbred lines, but the studies will be extended to commercial flocks. PMID:18817286

Kaiser, P; Howell, J; Fife, M; Sadeyen, J R; Salmon, N; Rothwell, L; Young, J; van Diemen, P; Stevens, M; Poh, T Y; Jones, M; Barrow, P; Swaggerty, C; Kogut, M; Smith, J; Burt, D



Carotenoids, sexual signals and immune function in barn swallows from Chernobyl.  


Carotenoids have been hypothesized to facilitate immune function and act as free-radical scavengers, thereby minimizing the frequency of mutations. Populations of animals exposed to higher levels of free radicals are thus expected to demonstrate reduced sexual coloration if use of carotenoids for free-radical scavenging is traded against use for sexual signals. The intensity of carotenoid-based sexual coloration was compared among three populations of barn swallows Hirundo rustica differing in exposure to radioactive contamination. Lymphocyte and immunoglobulin concentrations were depressed, whereas the heterophil:lymphocyte ratio, an index of stress, was enhanced in Chernobyl swallows compared to controls. Spleen size was reduced in Chernobyl compared to that of two control populations. Sexual coloration varied significantly among populations, with the size of a secondary sexual character (the length of the outermost tail feathers) being positively related to coloration in the two control populations, but not in the Chernobyl population. Thus the positive covariation between coloration and sexual signalling disappeared in the population subject to intense radioactive contamination. These findings suggest that the reliable signalling function of secondary sexual characters breaks down under extreme environmental conditions, no longer providing reliable information about the health status of males. PMID:10406129

Camplani, A; Saino, N; Møller, A P



Cardiac repair and regenerative potential in the goldfish (Carassius auratus) heart.  


The remarkable ability of the heart to regenerate has been demonstrated in the zebrafish and giant danio, two fish members of the cyprinid family. Here we use light and electron microscopy to examine the repair response in the heart of another cyprinid, the goldfish (Carassius auratus), following cautery injury to a small portion of its ventricular myocardium. We observed a robust inflammatory response in the first two weeks consisting primarily of infiltrating macrophages, heterophils, and melanomacrophages. These inflammatory cells were identified in the lumen of the spongy heart, within the site of the wound, and attached to endocardial cells adjacent to the site of injury. Marked accumulation of collagen fibers and increased connective tissue were also observed during the first and second weeks in a transition zone between healthy and injured myocardium as well as in adjacent sub-epicardial regions. The accumulation of collagen and connective tissue however did not persist. The presence of capillaries was also noted in the injured area during repair. The replacement of the cauterized region of the ventricle by myocardial tissue was achieved in 6weeks. The presence of ethynyl deoxyuridine-positive cardiac myocytes and partially differentiated cardiac myocytes during repair suggest effective cardiac myocyte driven regeneration mechanisms also operate in the injured goldfish heart, and are similar to those observed in zebrafish and giant danio. Our data suggest the ability for cardiac regeneration may be widely conserved among cyprinids. PMID:24548889

Grivas, Jamie; Haag, Maria; Johnson, Adedoyin; Manalo, Trina; Roell, Julia; Das, Tanmoy L; Brown, Evelyn; Burns, Alan R; Lafontant, Pascal J



Expression and adhesive ability of gicerin, a cell adhesion molecule, in the pock lesions of chorioallantoic membranes infected with an avian poxvirus.  

PubMed Central

The expression and adhesive activities of gicerin, a cell adhesion protein, in the pock lesions on chicken chorioallantoic membranes (CAM) infected with an avian poxvirus were studied. In normal CAMs, gicerin was found on the flattened epithelial cells, and neurite outgrowth factor (NOF) was in the basement membrane. However, in the pock lesions on infected CAMs, gicerin was overexpressed on the cell membranes of hyperplastic epithelial cells forming thick epithelial layers. Neurite outgrowth factor was also found mainly in the basement membrane, but occasionally showed aberrant expression among hyperplastic cells. In vitro analyses, using the dissociated cells from pock lesions, demonstrated that an anti-gicerin polyclonal antibody inhibit cell aggregation activity and cell adhesion to NOF. These results suggest that gicerin might promote the cell-cell and cell-extracellular matrix protein bindings of the hyperplastic epithelial cells by its homophilic and heterophilic adhesive activities, and contribute to pock formation on the infected CAMs. Images Figure 1. Figure 2. Figure 3. Figure 5.

Tsukamoto, Y; Kotani, T; Hiroi, S; Egawa, M; Ogawa, K; Sasaki, F; Taira, E



Mucocele in a spectacled owl (Pusilatrix perspicillata).  


A 6-year-old breeding female spectacled owl (Pusilatrix perspicillata) was presented for a soft, fluid-filled, spherical mass under the neck that had been increasing in size over the previous 3 days. Results of a fine-needle aspirate of the mass showed clear, pale-yellow fluid with a total protein of 12.6 g/L. Cytologic examination revealed erythrocytes, moderate numbers of heterophils, and numerous foamy mononuclear cells against a mucoid background. Macroscopically, the mass appeared to be attached firmly to the esophagus. The mass was excised surgically and submitted for histopathologic examination. The lesion comprised a circumscribed, fibrous-encapsulated multilocular cyst, lined by plump, goblet-type, cuboidal epithelial cells lying in abundant mucinous matrix. Findings were consistent with a mucocele of the esophageal mucosal gland. Excision was considered curative based on follow-up 6 months after initial presentation. To our knowledge, this is the first report of this condition in Strigiformes and indicates that mucocele should be included in the differential diagnosis of cervical masses in birds. PMID:24881153

Huynh, Minh; Brandão, João; Sabater, Mikel; Stidworthy, Mark F; Forbes, Neil A



Mycoplasma corogypsi associated polyarthritis and tenosynovitis in black vultures (Coragyps atratus)  

PubMed Central

Three wild American black vultures (Coragyps atratus) were presented to rehabilitation centers with swelling of multiple joints, including elbows, stifles, hocks, and carpal joints, and of the gastrocnemius tendons. Cytological examination of the joint fluid exudate indicated heterophilic arthritis. Radiographic examination in 2 vultures demonstrated periarticular soft tissue swelling in both birds and irregular articular surfaces with subchondral bone erosion in both elbows in 1 bird. Prolonged antibiotic therapy administered in 2 birds did not improve the clinical signs. Necropsy and histological examination demonstrated a chronic lymphoplasmacytic arthritis involving multiple joints and gastrocnemius tenosynovitis. Articular lesions varied in severity and ranged from moderate synovitis and cartilage erosion and fibrillation to severe synovitis, diffuse cartilage ulceration, subchondral bone loss and/or sclerosis, pannus, synovial cysts, and epiphyseal osteomyelitis. No walled bacteria were observed or isolated from the joints. However, mycoplasmas polymerase chain reactions were positive in at least 1 affected joint from each bird. Mycoplasmas were isolated from joints of 1 vulture that did not receive antibiotic therapy. Sequencing of 16S rRNA gene amplicons from joint samples and the mycoplasma isolate identified Mycoplasma corogypsi in 2 vultures and was suggestive in the third vulture. Mycoplasma corogypsi identification was confirmed by sequencing the 16S-23S intergenic spacer region of mycoplasma isolates. This report provides further evidence that M. corogypsi is a likely cause of arthritis and tenosynovitis in American black vultures. Cases of arthritis and tenosynovitis in New World vultures should be investigated for presence of Mycoplasma spp, especially M. corogypsi.

Van Wettere, A. J.; Ley, D. H.; Scott, D. E.; Buckanoff, H. D.; Degernes, L. A.



Diagnostic and clinical observation on the infectious bronchitis virus strain Q1 in Italy.  


This paper describes the diagnostic and clinical observations of an infectious bronchitis virus (IBV) variant, referred to as Q1, in clinically ill chickens in Italy. This IBV variant was described for the first time in 1998 in China. In the autumn of 2011 it caused a small-scale epidemic in nonvaccinated meat chickens in farms located in Northern Italy. The disease was characterized by increased mortality, kidney lesions and proventriculitis. Histopathological observations confirmed the nephritis and described an unusual erosive/necrotic proventriculitis with infiltration of lymphocytes, plasma cells and heterophils, as well as fibroplasia in the lamina propria. Despite these findings and the isolation of the Q1 IB virus directly from proventricular tissue, further studies are necessary to confirm the role of this IBV strain in the development of proventricular lesions. Phylogenetic analysis revealed that all the IBV isolates were very similar and probably had a common origin. The IBV Q1 variant appears to be now endemic in the North of Italy and at times it is detected in vaccinated backyard and commercial broiler farms. The importance of continuous monitoring in controlling the spread of known or emerging IBV variants is underlined. PMID:24362775

Toffan, Anna; Bonci, Michela; Bano, Luca; Bano, Luca; Valastro, Viviana; Vascellari, Marta; Capua, Ilaria; Terregino, Calogero



The receptor tyrosine kinase ARK mediates cell aggregation by homophilic binding.  

PubMed Central

The ARK (AXL, UFO) receptor is a member of a new family of receptor tyrosine kinases whose extracellular domain contains a combination of fibronectin type III and immunoglobulin motifs similar to those found in many cell adhesion molecules. ARK mRNA is expressed at high levels in the mouse brain, prevalently in the hippocampus and cerebellum, and this pattern of expression resembles that of adhesion molecules that are capable of promoting cell aggregation through homophilic or heterophilic binding. We report here the ability of the murine ARK receptor to mediate homophilic binding. Expression of the ARK protein in Drosophila S2 cells induces formation of cell aggregates consisting of ARK-expressing cells, and aggregation leads to receptor activation, with an increase in receptor phosphorylation. Homophilic binding does not require ARK tyrosine kinase activity, since S2 cells expressing a receptor in which the intracellular domain was deleted were able to undergo aggregation as well as cells expressing the wild-type ARK receptor. Similar results were obtained with NIH 3T3 and CHO cells expressing high levels of ARK, although in this case ARK expression appeared to be accompanied by constitutive activation. The purified recombinant extracellular domain of ARK can induce homotypic aggregation of coated fluorescent beads (Covaspheres), and this protein can also function as a substrate for adhesion by S2 and NIH 3T3 cells expressing ARK. These results suggest that ARK represents a new cell adhesion molecule that through its homophilic interaction may regulate cellular functions during cell recognition.

Bellosta, P; Costa, M; Lin, D A; Basilico, C



Adhesion and nanomechanics of pili from the probiotic Lactobacillus rhamnosus GG.  


Knowledge of the mechanisms by which bacterial pili adhere to host cells and withstand external forces is critical to our understanding of their functional roles and offers exciting avenues in biomedicine for controlling the adhesion of bacterial pathogens and probiotics. While much progress has been made in the nanoscale characterization of pili from Gram-negative bacteria, the adhesive and mechanical properties of Gram-positive bacterial pili remain largely unknown. Here, we use single-molecule atomic force microscopy to unravel the binding mechanism of pili from the probiotic Gram-positive bacterium Lactobacillus rhamnosus GG (LGG). First, we show that SpaC, the key adhesion protein of the LGG pilus, is a multifunctional adhesin with broad specificity. SpaC forms homophilic trans-interactions engaged in bacterial aggregation and specifically binds mucin and collagen, two major extracellular components of host epithelial layers. Homophilic and heterophilic interactions display similar binding strengths and dissociation rates. Next, pulling experiments on living bacteria demonstrate that LGG pili exhibit two unique mechanical responses, that is, zipper-like adhesion involving multiple SpaC molecules distributed along the pilus length and nanospring properties enabling pili to resist high force. These mechanical properties may represent a generic mechanism among Gram-positive bacterial pili for strengthening adhesion and withstanding shear stresses in the natural environment. The single-molecule experiments presented here may help us to design molecules capable of promoting or inhibiting bacterial-host interactions. PMID:23531039

Tripathi, Prachi; Beaussart, Audrey; Alsteens, David; Dupres, Vincent; Claes, Ingmar; von Ossowski, Ingemar; de Vos, Willem M; Palva, Airi; Lebeer, Sarah; Vanderleyden, Jos; Dufrêne, Yves F



Maintenance of neuronal positions in organized ganglia by SAX-7, a Caenorhabditis elegans homologue of L1.  


The L1 family of cell adhesion molecules is predominantly expressed in the nervous system. Mutations in human L1 cause neuronal diseases such as HSAS, MASA, and SPG1. Here we show that sax-7 gene encodes an L1 homologue in Caenorhabditis elegans. In sax-7 mutants, the organization of ganglia and positioning of neurons are abnormal in the adult stage, but these abnormalities are not observed in early larval stage. Misplacement of neurons in sax-7 mutants is triggered by mechanical force linked to body movement. Short and long forms of SAX-7 exhibited strong and weak homophilic adhesion activities in in vitro aggregation assay, respectively, which correlated with their different activities in vivo. SAX-7 was localized on plasma membranes of neurons in vivo. Expression of SAX-7 only in a single neuron in sax-7 mutants cell-autonomously restored its normal neuronal position. Expression of SAX-7 in two different head neurons in sax-7 mutants led to the forced attachment of these neurons. We propose that both homophilic and heterophilic interactions of SAX-7 are essential for maintenance of neuronal positions in organized ganglia. PMID:15775964

Sasakura, Hiroyuki; Inada, Hitoshi; Kuhara, Atsushi; Fusaoka, Eri; Takemoto, Daisuke; Takeuchi, Kosei; Mori, Ikue



A genetically encoded tag for correlated light and electron microscopy of intact cells, tissues, and organisms.  


Electron microscopy (EM) achieves the highest spatial resolution in protein localization, but specific protein EM labeling has lacked generally applicable genetically encoded tags for in situ visualization in cells and tissues. Here we introduce "miniSOG" (for mini Singlet Oxygen Generator), a fluorescent flavoprotein engineered from Arabidopsis phototropin 2. MiniSOG contains 106 amino acids, less than half the size of Green Fluorescent Protein. Illumination of miniSOG generates sufficient singlet oxygen to locally catalyze the polymerization of diaminobenzidine into an osmiophilic reaction product resolvable by EM. MiniSOG fusions to many well-characterized proteins localize correctly in mammalian cells, intact nematodes, and rodents, enabling correlated fluorescence and EM from large volumes of tissue after strong aldehyde fixation, without the need for exogenous ligands, probes, or destructive permeabilizing detergents. MiniSOG permits high quality ultrastructural preservation and 3-dimensional protein localization via electron tomography or serial section block face scanning electron microscopy. EM shows that miniSOG-tagged SynCAM1 is presynaptic in cultured cortical neurons, whereas miniSOG-tagged SynCAM2 is postsynaptic in culture and in intact mice. Thus SynCAM1 and SynCAM2 could be heterophilic partners. MiniSOG may do for EM what Green Fluorescent Protein did for fluorescence microscopy. PMID:21483721

Shu, Xiaokun; Lev-Ram, Varda; Deerinck, Thomas J; Qi, Yingchuan; Ramko, Ericka B; Davidson, Michael W; Jin, Yishi; Ellisman, Mark H; Tsien, Roger Y



Proventricular dilatation disease associated with Avian bornavirus in a scarlet macaw (Ara macao).  


A case of proventricular dilatation disease is described in a scarlet macaw (Ara macao) from clinical presentation to diagnosis with molecular methods. The initial clinical signs were depression progressing to head pressing over several days. A leukocytosis with toxic heterophil changes, hypoalbuminemia, and increased serum activity of aspartate aminotransferase and creatine kinase were present. Lead and zinc assays were within reference ranges, and results from Chlamydophila and polyomavirus testing were negative. Contrast-enhanced fluoroscopy revealed normal gastrointestinal transit times and motility as well as the presence of 2 small metallic foreign bodies in the ventriculus. The macaw was treated with antimicrobials, analgesics, vitamins E and B complex, force-feeding, and fluid administration with little improvement. Euthanasia was elected, and histologic examination of brain tissue revealed a perivascular lymphoplasmacytic infiltration, while the lungs had evidence of a fungal pneumonia. Tissue samples from the brain and proventriculus tested positive for the presence of Avian bornavirus genotype 2, while serology confirmed Avian bornavirus infection. PMID:21088184

Keller, Dominique L; Honkavuori, Kirsi S; Briese, Thomas; Lipkin, W Ian; Muthuswamy, Anantharaman; Steinberg, Howard; Sladky, Kurt K



Riemerella anatipestifer outbreaks in commercial goose flocks and identification of isolates by MALDI-TOF mass spectrometry.  


Several outbreaks of Riemerella anatipestifer in commercial geese occurred within a short time period. A serious disease was recognized in the affected birds, mainly characterized by depression and severe neurologic disturbances. The morbidity ranged from 20 to 30% and the mortality from 5 to 20%. Generally, the clinical signs started at the age of 8 to 10 days. Post-mortem examination revealed fibrinous pericarditis, perihepatitis and airsacculitis in all birds. Some of the birds also had synovitis of the tibio-tarsal joints and oedematous swelling of the subcutaneous tissues around these joints and metatarsus. Histology revealed a characteristic severe inflammation with heterophilic granulocytes in different organs. Bacteriological investigation was made from several organs and R. anatipestifer could be isolated from all birds investigated. The identification of these clinical isolates, done for the first time by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry, confirmed the aetiology. Sequence analysis showed 100% similarity between the clinical isolates, indicating a common source of infection. PMID:23581442

Hess, C; Enichlmayr, H; Jandreski-Cvetkovic, D; Liebhart, D; Bilic, I; Hess, M



Pro-inflammatory effects of uric acid in the gastrointestinal tract  

PubMed Central

Uric acid can be generated in the gastrointestinal (GI) tract from the breakdown of nucleotides ingested in the diet or from purines released from host cells as a result of pathogen-induced cell damage. Xanthine oxidase (XO) is the enzyme that converts hypoxanthine or xanthine into uric acid, a reaction that also generates hydrogen peroxide. It has been assumed that the product of XO responsible for the pro-inflammatory effects of this enzyme is hydrogen peroxide. Recent literature on uric acid, however, has indicated that uric acid itself may have biological effects. We tested whether uric acid itself has detectable pro-inflammatory effects using an in vivo model using ligated rabbit intestinal segments (“loops”) as well as in vitro assays using cultured cells. Addition of exogenous uric acid increased the influx of heterophils into rabbit intestinal loops, as measured by myeloperoxidase activity. In addition, white blood cells adhered avidly to uric acid crystals, forming large aggregates of cells. Uric acid acts as a leukocyte chemoattractant in the GI tract. The role of uric acid in enteric infections and in non-infectious disorders of the GI tract deserves more attention.

Crane, John K.; Mongiardo, Krystin M.



Assessment of the stresses imposed on adult ostriches (Struthio camelus) during handling, loading, transportation and unloading.  


The stresses imposed during the handling, loading and unloading of 250 adult ostriches (Struthio camelus) transported by road were evaluated, weighted, scored and later compared with some objective physiological indices of stress measured after the journey. During handling, the numbers of slips and falls, incidents of aggressive behaviour, the calculated behavioural points, the number of injuries recorded per ostrich and the time spent were significantly (P<0.01) greater than the values recorded during loading and unloading. During handling and loading, 45 per cent of the ostriches had a good score (1.1 to 2 points), 15.5 per cent had a fair score (2.1 to 3 points) and 39.4 per cent had a poor or bad score (more than 3 points). The behavioural scores were significantly and positively correlated with the heterophil:lymphocyte ratio, the rectal temperature and the number of injuries sustained by the ostriches. The results showed that the poorer the behavioural score, the higher the level of stress suffered by the ostriches during handling and loading. PMID:18587061

Minka, N S; Ayo, J O



Evidence of a dominance hierarchy in captive Caribbean flamingos and its relation to pair bonding and physiological measures of health.  


Caribbean flamingo social structure, how pair bonds affect the structure of the flock, and how social stress affects health measured by heterophil to lymphocyte ratios (H/L) were investigated at the Philadelphia Zoo. It was hypothesized that a hierarchy may become apparent by analyzing agonistic interactions and that paired individuals would share similar places within the hierarchy. Furthermore, it was hypothesized that a negative relationship between H/L ratio and dominance would exist. Forty observations were conducted and in 70% of interactions instigating bird(s) won the encounter, suggesting either some advantage for instigating birds or a prior expectation of an encounter's outcome based upon an understanding of the flock's hierarchy. The flock possessed a semi-linear hierarchy (in terms of wins/losses) and birds with higher pair-bond strengths maintained dominant positions, suggesting that pair-bonding may help individuals become more successful in agonistic encounters. Birds who won more often had higher lymphocytes percentages and analyses suggested a trend indicating dominant birds may be less stressed. A semi-linear hierarchy was also found in terms of initiation/being targeted, and a bird's rank on the dominance (wins/losses) and initiate/target hierarchies were positively correlated, suggesting that subdominant birds were targeted by dominant birds more frequently than vice versa. PMID:24670555

Royer, Erica A; Anderson, Matthew J



Clinical and laboratory presentation of EBV positive infectious mononucleosis in young adults.  

PubMed Central

Clinical descriptions of Epstein-Barr virus (EBV) positive infectious mononucleosis (IM) are rare and their results are inconsistent. Over a 4-year period, we prospectively studied 590 young adults with clinically suspected IM, all of whom were tested for the presence of EBV IgM antibodies. We investigated the demographical, clinical and laboratory features of subjects with positive EBV IgM serology and heterophile antibodies. Contrary to previous studies, we found a seasonal disease pattern with a peak incidence during summer months, and a lower-than-expected prevalence of lymphadenopathy (88.9%), leucocytosis (46.2%), atypical lymphocytosis (89.2%) and elevated liver enzymes (57.9%). The prevalence of hyperbilirubinemia was relatively high (14.9%). The classic triad of fever, sore throat and lymph-adenopathy had relatively low sensitivity (68.2%) and specificity (41.9%) for EBV infection. Our study provides a complete and updated description of the clinical and laboratory presentation of laboratory confirmed IM, which is important for both clinicians and epidemiologists.

Grotto, I.; Mimouni, D.; Huerta, M.; Mimouni, M.; Cohen, D.; Robin, G.; Pitlik, S.; Green, M. S.



The CD2 family of natural killer cell receptors.  


The CD2 family of receptors is evolutionarily conserved and widely expressed on cells within the hematopoietic compartment. In recent years several new members have been identified with important roles in the immune system. CD2 family members regulate natural killer (NK) cell lytic activity and inflammatory cytokine production when engaged by ligands on tumor cells. Furthermore, a subfamily of CD2 receptors, the CD 150-like molecules, has been implicated in the pathogenesis of X-linked lymphoproliferative disease (XLP). Many of these receptors have now been shown to bind homophilically or heterophilically to other molecules within the family. With these discoveries a novel mechanism for lymphocyte regulation has emerged: CD2 family members on NK cells engage ligands on neighboring NK cells, leading to NK cell stimulation. Moreover, heterotypic stimulatory interactions between NK cells and other leukocytes also occur. In this manner, CD2 family members may provide interlymphocyte communication that maintains organization within the hematopoietic compartment and amplifies immune responses. This review discusses these multiple roles for CD2 family members, focusing specifically on the regulation of NK cells. PMID:16323413

McNerney, M E; Kumar, V



E-cadherin-mediated interactions of thymic epithelial cells with CD103+ thymocytes lead to enhanced thymocyte cell proliferation.  


Cadherins are a family of cell adhesion molecules that mainly mediate homotypic homophilic interactions, but for E-cadherin, heterophilic interactions with the integrin alpha(E)(CD103)beta(7) have also been reported. In the human thymus, where thymocytes develop in close contact with thymic stromal cells, E-cadherin expression was detected on thymic epithelial cells. By immunofluorescence staining, the strongest expression of E-cadherin was observed on medullary thymic epithelial cells. These cells also express cytosolic catenins, which are necessary to form functional cadherin-catenin complexes. Regardless of their developmental stage, human thymocytes do not express E-cadherin, indicating that homophilic interactions cannot occur. Flow cytometric analysis revealed that the E-cadherin ligand CD103 is expressed on subpopulations of the early CD4(-) CD8(-) double-negative and of the more mature CD8(+) single-positive thymocytes. Using an in vitro cell adhesion assay, double-negative and CD8(+) single-positive thymocytes adhered strongly to isolated thymic epithelial cells. These adhesive interactions could be inhibited by antibodies against E-cadherin or CD103. CD8(+) thymocytes showed a proliferative response when incubated with thymic epithelial cells. This mitogenic effect was inhibited by antibodies against CD103, which strongly indicates a direct involvement of the adhesive ligand pair CD103-E-cadherin in human thymocyte cell proliferation. PMID:12414996

Kutlesa, Snjezana; Wessels, Johannes T; Speiser, Angelika; Steiert, Inge; Müller, Claudia A; Klein, Gerd



New models of chronic synovitis in rabbits induced by mycoplasmas: microbiological, histopathological, and immunological observations on rabbits injected with Mycoplasma arthritidis and Mycoplasma pulmonis.  

PubMed Central

A dose-dependent chronic synovitis was induced in rabbit knees after the intra-articular injection of both Mycoplasma arthritidis and Mycoplasma pulmonis. The inflammation progressed from an initial acute phase at 1 week characterized by edema, infiltration of the synovium with monocytes and heterophils, and desquamation of lining cells, to a more chronic phase at 1 and 3 months, in which villus hyperplasia, lymph "nodules," mononuclear cell infiltration, fibroplasia, and collagen deposition were prominent. With one exception, mycoplasmas could no longer be cultivated from the joints 1 month postinoculation. Both mycoplasma species evoked a humoral antibody response that was more marked in synovial fluids than in peripheral blood. A cell-mediated immune reaction, as evidence by enhanced uptake by [3H]thymidine by sensitized blood, spleen, or node lymphocytes in the presence of homologous antigen, was detected only in rabbits injected with M. pulmonis. Lymphocytes taken from arthritic rabbits were no more cytotoxic toward synovial cells derived from normal or arthritic rabbits than were normal lymphocytes. The models of synovitis described in this study offer a convenient probe for determining the mechanisms of mycoplasma-induced inflammation, since they require only a single injection of the initiating agent and, in addition, utilize an animal host large enough for detailed investigation into the nature of mycoplasma/synovium interactions. Images

Cole, B C; Griffiths, M M; Eichwald, E J; Ward, J R



Modulatory effect of ascorbic acid on physiological responses of transported ostrich chicks.  


The aims of this study were to determine the modulating role of ascorbic acid (AA) on rectal temperature (RT), heterophil to lymphocyte (H to L) ratio and aberrant behaviours of ostrich chicks transported by road for 4 h during hot-dry conditions. Twenty ostrich chicks aged 2.5 months, of both sexes and belonging to the Red Neck breed, served as subjects of the study. The chicks were assigned randomly to AA-treated and control groups, consisting of 10 chicks each. The AA-treated group was administered orally with 100 mg/kg body weight of AA dissolved in 5 mL of sterile water 30 min before transportation, whilst the control group was given the equivalent of sterile water only. The thermal load (TL) experienced in the vehicle during transportation fluctuated between 31 °C and 89 °C, as calculated from the ambient temperature and relative humidity. Transportation induced hyperthermia, lymphopenia, heterophilia and aberrant behaviours of pecking, wing fluffing and panting, which were ameliorated by AA administration. The relationships between the TL, journey duration and physiological variables of RT, H to L ratio and aberrant behaviours recorded during transportation were significantly and positively correlated in the control group. In AA-treated group the relationships were not significantly correlated. In conclusion, the results showed for the first time that AA ameliorated the adverse effects of stress caused by road transportation on the aberrant behaviours, RT and H to L ratio of ostrich chicks during the hot-dry season. PMID:23327313

Salka, Minka N; Olusegun, Ayo J



Superoxide dismutase expression and oxidative damage in a case of myopathy in brown pelicans (Pelecanus occidentalis).  


Four brown pelicans (Pelecanus occidentalis) housed at a rehabilitation facility were found dead after a 3-day history of muscle weakness and after being fed for about 2 weeks from a recent shipment of fish. The birds had pale streaking of the skeletal and heart muscles. Microscopically, the skeletal muscle, and to a lesser extent the cardiac muscle, had severe myocyte degeneration and necrosis characterized by microvacuolation with loss of cross-striations, condensation of cytoplasm, fragmentation, mineralization, and inflammatory cell infiltrates consisting of multinucleated cells, macrophages, and few heterophils. The findings were consistent with myopathy, and a nutritional myopathy caused by eating rancid fish was suspected. Immunohistochemical staining revealed abundant immunoreactive copper zinc superoxide dismutase and manganese superoxide dismutase either as diffuse homogeneous precipitates or granular aggregates in the cytoplasm of affected cells. Immunoreactivity was directly related to degree of cellular damage as estimated by light microscopic examination. We suggest that the lack of protection, despite upregulation of superoxide dismutase, is most likely attributable to supersaturation of oxidants beyond the capacity of superoxide dismutases to scavenge. PMID:17459863

Giri, Dipak K; Miller, Debra L; Thompson, Larry J; Mailler, Lesley; Styer, Eloise; Baldwin, Charles



Identification of a novel collagen type ?-binding protein from Streptococcus suis serotype 2.  


Streptococcus suis, a major pathogen of pigs, is an emerging zoonotic agent that causes meningitis and septic shock. cbp40 is a putative virulent gene that has been identified using suppression subtractive hybridization performed on the virulent S. suis serotype 2 strain HA9801 and the avirulent S. suis serotype 2 strain T15. Based on predicted protein features showing a shared conserved domain with the collagen-binding protein Cna of Staphylococcus aureus, Cbp40 is likely to function as a direct mediator of collagen adhesion. Here, the cbp40 gene was cloned and the recombinant protein purified. Western blotting using swine convalescent sera confirmed its role as an immunogenic protein. Collagen binding activity could be detected by western affinity blot and ELISA. Conversely, deletion of the cbp40 gene reduced bacterial adhesion to HEp-2 cells, capacity for biofilm formation, and virulence in a zebrafish infection model. The response of the bEnd.3 cell line to infection with the S. suis serotype 2 strain ZY05719 and the cbp40-knockout strain was evaluated using gene expression arrays. The differentially expressed genes were involved in inflammatory and immune responses, leukocyte adhesion and heterophilic cell adhesion. Collectively, these data suggest that Cbp40 plays an important role as an extracellular matrix adhesion protein that interacts with host cells during infection. PMID:23465548

Zhang, Hui; Ma, Zhe; Li, Yue; Zheng, Junxi; Yi, Li; Fan, Hongjie; Lu, Chengping



Intestinal brush border assembly driven by protocadherin-based intermicrovillar adhesion.  


Transporting epithelial cells build apical microvilli to increase membrane surface area and enhance absorptive capacity. The intestinal brush border provides an elaborate example with tightly packed microvilli that function in nutrient absorption and host defense. Although the brush border is essential for physiological homeostasis, its assembly is poorly understood. We found that brush border assembly is driven by the formation of Ca(2+)-dependent adhesion links between adjacent microvilli. Intermicrovillar links are composed of protocadherin-24 and mucin-like protocadherin, which target to microvillar tips and interact to form a trans-heterophilic complex. The cytoplasmic domains of microvillar protocadherins interact with the scaffolding protein, harmonin, and myosin-7b, which promote localization to microvillar tips. Finally, a mouse model of Usher syndrome lacking harmonin exhibits microvillar protocadherin mislocalization and severe defects in brush border morphology. These data reveal an adhesion-based mechanism for brush border assembly and illuminate the basis of intestinal pathology in patients with Usher syndrome. PAPERFLICK: PMID:24725409

Crawley, Scott W; Shifrin, David A; Grega-Larson, Nathan E; McConnell, Russell E; Benesh, Andrew E; Mao, Suli; Zheng, Yuxi; Zheng, Qing Yin; Nam, Ki Taek; Millis, Bryan A; Kachar, Bechara; Tyska, Matthew J



Immunostimulatory properties of Toll-like receptor ligands in chickens.  


Toll-like receptors (TLRs) are evolutionarily conserved pattern recognition receptors that have been identified in mammals and avian species. Ligands for TLRs are typically conserved structural motifs of microorganisms termed pathogen-associated molecular patterns (PAMPs). Several TLRs have been detected in many cell subsets, such as in macrophages, heterophils and B cells, where they mediate host-responses to pathogens by promoting cellular activation and the production of cytokines. Importantly, TLR ligands help prime a robust adaptive immune response by promoting the maturation of professional antigen presenting cells. These properties make TLR ligands an attractive approach to enhance host-immunity to pathogens by administering them either prophylactically or in the context of a vaccine adjuvant. In this review, we discuss what is known about the immunostimulatory properties of TLR ligands in chickens, both at the cellular level as well as in vivo. Furthermore, we highlight previous successes in exploiting TLR ligands to protect against several pathogens including avian influenza virus, Salmonella, Escherichia coli, and Newcastle disease Virus. PMID:23305711

St Paul, Michael; Brisbin, Jennifer T; Abdul-Careem, Mohamed Faizal; Sharif, Shayan



Low temperatures reduce skin healing in the Jacar? do Pantanal (Caiman yacare, Daudin 1802)  

PubMed Central

Summary Studies of skin wound healing in crocodilians are necessary given the frequent occurrence of cannibalism in intensive farming systems. Air temperature affects tissue recovery because crocodilians are ectothermic. Therefore, the kinetics of skin wound healing in Caiman yacare were examined at temperatures of 33°C and 23°C. Sixteen caiman were selected and divided into two groups of eight maintained at 23°C or 33°C. The studied individuals' scars were photographed after 1, 2, 3, 7, 15 and 30 days of the experimental conditions, and samples were collected for histological processing after 3, 7, 15 and 30 days. Macroscopically, the blood clot (heterophilic granuloma) noticeably remained in place covering the wound longer for the caiman kept at 23°C. Microscopically, the temperature of 23°C slowed epidermal migration and skin repair. Comparatively, new blood vessels, labeled using von Willebrand factor (vWF) antibody staining, were more frequently found in the scars of the 33°C group. The collagen fibers in the dermis were denser in the 33°C treatment. Considering the delayed healing at 23°C, producers are recommended to keep wounded animals at 33°C, especially when tanks are cold, to enable rapid wound closure and better repair of collagen fibers because such lesions tend to compromise the use of their skin as leather.

Pressinotti, Leandro Nogueira; Borges, Ricardo Moraes; Alves De Lima, Angela Paula; Aleixo, Victor Manuel; Iunes, Renata Stecca; Borges, Joao Carlos Shimada; Cogliati, Bruno; Cunha Da Silva, Jose Roberto Machado



Influence of chromium propionate on the carcass characteristics and immune response of commercial broiler birds under normal rearing conditions.  


The study investigated the effect of supplementation of chromium propionate at different dosage levels (100 to 3,200 µg/kg of elemental chromium in feed) on the performance, carcass characteristics, and immune response of broiler birds. The study was conducted on male broiler chickens (Cobb 400) for a period of 42 d as per the completely randomized design. Seven hundred 1-d-old birds were randomly segregated into 7 treatment groups, each with 10 replicates, and each replicate having 10 birds. Weight gain, feed intake, carcass characteristics, immune response, and the serum biochemical parameters of birds were studied during the supplementation period. There was no significant response to chromium supplementation on weight gain, feed intake, FCR, and lymphoid organ weights for the overall study period of 42 d. With increased chromium dosage, the breast meat yield improved linearly (P = 0.045). Antibody response to Newcastle disease vaccination improved quadratically (P = 0.001) with increased dose of chromium in the diet. Improved cell-mediated immune response was indicated by the increase (quadratic, P = 0.00) in lymphocyte proliferation ratio with increased dose of chromium supplementation. Heterophil:lymphocyte ratio decreased (quadratic, P = 0.004) with chromium propionate dosage, suggestive of reduced stress levels. Chromium propionate supplementation also reduced serum glucose levels (quadratic, P = 0.008) and improved (quadratic, P = 0.016) the total protein levels. PMID:24604850

Rajalekshmi, M; Sugumar, C; Chirakkal, H; Ramarao, S V



Hematology and blood chemistry reference values and age-related changes in wild Bearded Vultures (Gypaetus barbatus).  


Normal hematologic and blood chemistry values for clinical use and age-related changes are reported as reference values for the endangered Bearded Vulture (Gypaetus barbatus). Blood samples were obtained from 21 nestlings and 26 free-living subadults and adults. No significant differences were found between subadults and adults or between sexes for any of the studied parameters. Reference ranges have been established for Bearded Vulture nestlings (less than 3 mo of age) and for free-living Bearded Vultures, with subadult and adult data combined without affecting clinical interpretation. Some reference values for the parameters reported in this study are similar to those previously described for vultures and other raptor species, although creatine phosphokinase and lactate dehydrogenase activities were higher than those reported for birds of prey. Significant age-related differences were identified in urea, uric acid, triglycerides, total serum protein, inorganic phosphorus, and magnesium concentrations, as well as aspartate aminotransferase, creatine phosphokinase, lactate dehydrogenase, alkaline phosphatase, amylase, and lipase activities (P<0.05). Additionally, significant age-related differences were noted in red and white blood cell counts, packed cell volume, hemoglobin concentration, mean corpuscular volume, mean corpuscular hemoglobin concentration, fibrinogen level, and heterophils, lymphocytes, and eosinophils (P<0.005). The results obtained from this study provide reference ranges that will be useful for evaluating the pathologic conditions and general health of Bearded Vulture populations and reveal the existence of important age-related differences in the species. PMID:20688632

Hernández, M; Margalida, A



Mammalian Fat and Dachsous cadherins regulate apical membrane organization in the embryonic cerebral cortex  

PubMed Central

Compartmentalization of the plasma membrane in a cell is fundamental for its proper functions. In this study, we present evidence that mammalian Fat4 and Dachsous1 cadherins regulate the apical plasma membrane organization in the embryonic cerebral cortex. In neural progenitor cells of the cortex, Fat4 and Dachsous1 were concentrated together in a cell–cell contact area positioned more apically than the adherens junction (AJ). These molecules interacted in a heterophilic fashion, affecting their respective protein levels. We further found that Fat4 associated and colocalized with the Pals1 complex. Ultrastructurally, the apical junctions of the progenitor cells comprised the AJ and a stretch of plasma membrane apposition extending apically from the AJ, which positionally corresponded to the Fat4–Dachsous1-positive zone. Depletion of Fat4 or Pals1 abolished this membrane apposition. These results highlight the importance of the Fat4–Dachsous1–Pals1 complex in organizing the apical membrane architecture of neural progenitor cells.

Ishiuchi, Takashi; Misaki, Kazuyo; Yonemura, Shigenobu; Tanoue, Takuji



Chronic myelogenous leukemia in a great horned owl (Bubo virginianus).  


A free-ranging adult female great horned owl (Bubo virginianus) was presented to the Wildlife Medical Clinic at the University of Illinois after being observed with anorexia and decreased activity. A severe leukocytosis (212 400 cells/microl), primarily comprised of mature heterophils, was found at presentation. Results of various diagnostic tests including radiographs, Chlamydophila serologic testing, measurement of Aspergillus antibody and antigen titers, plasma protein electrophoresis, fecal culture and acid-fast staining, coelioscopy, endoscopy, tracheoscopy, exploratory coelomotomy, nuclear scintigraphy, tissue cultures, bone marrow biopsy, and histopathology revealed no underlying cause for the persistent leukocytosis. No response to treatment with antibiotics or antifungal agents was observed, although a transient, significant decrease in the leukocyte count (6200 cells/microl) was observed after treatment with fenbendazole. A presumptive diagnosis of chronic myelogenous leukemia was made based on 3 factors: disease duration of greater than 3 months, a lack of identifiable foci of inflammation, and a lack of response to conventional therapy. The diagnosis was confirmed based on postmortem examination and testing 177 days after initial presentation. PMID:19530405

Wiley, Jennifer L; Whittington, Julia K; Wilmes, Christine M; Messick, Joanne B



Transfusion-associated cytomegalovirus mononucleosis.  

PubMed Central

Transfusion-associated cytomegalovirus mononucleosis is generally considered only as a complication of extracorporeal circulation following cardiac surgery. Three cases following trauma were recognized in less than one year. Both massive and limited volume blood transfusions were involved. Hectic fever was a characteristic feature in these otherwise remarkably asymptomatic individuals, without the classic features of heterophile-positive infectious mononucleosis. Since the illness developed several weeks into the post-operative period after extensive thoracic or abdominal trauma surgery, the presence of an undrained abscess was naturally the major diagnostic concern. Atypical lymphocytosis, markers of altered immunity (cold agglutinins, rheumatoid factor) and moderate hepatic dysfunction were important laboratory clues. In one case, focal isotope defects in the spleen scan misleadingly suggested a septic complication. A false-positive monospot test initially obscured the correct serologic diagnosis in the same patient. Failure to consider this selflimited viral infection may be a critical factor leading to unnecessary surgery. Other viral agents capable of eliciting a similar syndrome are cited. Images Fig. 1.

Lerner, P I; Sampliner, J E



AMIGO, a transmembrane protein implicated in axon tract development, defines a novel protein family with leucine-rich repeats.  


Ordered differential display identified a novel sequence induced in neurons by the neurite-promoting protein amphoterin. We named this gene amphoterin-induced gene and ORF (AMIGO), and also cloned two other novel genes homologous to AMIGO (AMIGO2 and AMIGO3). Together, these three AMIGOs form a novel family of genes coding for type I transmembrane proteins which contain a signal sequence for secretion and a transmembrane domain. The deduced extracellular parts of the AMIGOs contain six leucine-rich repeats (LRRs) flanked by cysteine-rich LRR NH2- and COOH-terminal domains and by one immunoglobulin domain close to the transmembrane region. A substrate-bound form of the recombinant AMIGO ectodomain promoted prominent neurite extension in hippocampal neurons, and in solution, the same AMIGO ectodomain inhibited fasciculation of neurites. A homophilic and heterophilic binding mechanism is shown between the members of the AMIGO family. Our results suggest that the members of the AMIGO protein family are novel cell adhesion molecules among which AMIGO is specifically expressed on fiber tracts of neuronal tissues and participates in their formation. PMID:12629050

Kuja-Panula, Juha; Kiiltomäki, Marjaana; Yamashiro, Takashi; Rouhiainen, Ari; Rauvala, Heikki



AMIGO, a transmembrane protein implicated in axon tract development, defines a novel protein family with leucine-rich repeats  

PubMed Central

Ordered differential display identified a novel sequence induced in neurons by the neurite-promoting protein amphoterin. We named this gene amphoterin-induced gene and ORF (AMIGO), and also cloned two other novel genes homologous to AMIGO (AMIGO2 and AMIGO3). Together, these three AMIGOs form a novel family of genes coding for type I transmembrane proteins which contain a signal sequence for secretion and a transmembrane domain. The deduced extracellular parts of the AMIGOs contain six leucine-rich repeats (LRRs) flanked by cysteine-rich LRR NH2- and COOH-terminal domains and by one immunoglobulin domain close to the transmembrane region. A substrate-bound form of the recombinant AMIGO ectodomain promoted prominent neurite extension in hippocampal neurons, and in solution, the same AMIGO ectodomain inhibited fasciculation of neurites. A homophilic and heterophilic binding mechanism is shown between the members of the AMIGO family. Our results suggest that the members of the AMIGO protein family are novel cell adhesion molecules among which AMIGO is specifically expressed on fiber tracts of neuronal tissues and participates in their formation.

Kuja-Panula, Juha; Kiiltomaki, Marjaana; Yamashiro, Takashi; Rouhiainen, Ari; Rauvala, Heikki



In vivo magnetic resonance imaging and spectroscopy identifies oncolytic adenovirus responders.  


At present, it is not possible to reliably identify patients who will benefit from oncolytic virus treatments. Conventional modalities such as computed tomography (CT), which measure tumor size, are unreliable owing to inflammation-induced tumor swelling. We hypothesized that magnetic resonance imaging (MRI) and spectroscopy (MRS) might be useful in this regard. However, little previous data exist and neither oncolytic adenovirus nor immunocompetent models have been assessed by MRS. Here, we provide evidence that in T2-weighted MRI a hypointense core area, consistent with coagulative necrosis, develops in immunocompetent Syrian hamster carcinomas that respond to oncolytic adenovirus treatment. The same phenomenon was observed in a neuroblastoma patient while he responded to the treatment. With relapse at a later stage, however, the tumor of this patient became moderately hyperintense. We found that MRS of taurine, choline and unsaturated fatty acids can be useful early indicators of response and provide detailed information about tumor growth and degeneration. In hamsters, calprotectin-positive inflammatory cells (heterophils and macrophages) were found in abundance; particularly surrounding necrotic areas in carcinomas and T cells were significantly increased in sarcomas, when these had been treated with a granulocyte-macrophage colony-stimulating factor-producing virus, suggesting a possible link between oncolysis, necrosis (seen as a hypointense core in MRI) and/or immune response. Our study indicates that both MRI and MRS could be useful in the estimation of oncolytic adenovirus efficacy at early time points after treatment. PMID:24248808

Hemminki, O; Immonen, R; Närväinen, J; Kipar, A; Paasonen, J; Jokivarsi, K T; Yli-Ollila, H; Soininen, P; Partanen, K; Joensuu, T; Parvianen, S; Pesonen, S K; Koski, A; Vähä-Koskela, M; Cerullo, V; Pesonen, S; Gröhn, O H; Hemminki, A



Physiological indices of stress in wild and captive garter snakes: Correlations, repeatability, and ecological variation.  


Glucocorticoids and leukocyte ratios have become the most widespread variables employed to test hypotheses regarding physiological stress in wild and captive vertebrates. Little is known, however, regarding how these two indices of stress covary in response to stressors, their repeatability within individuals, and differences in response time upon capture. Furthermore, few studies compare stress indices between captive and wild populations, to assess potential alteration of stress physiology in captivity. To address these issues, we examined corticosterone (CORT) and heterophil to lymphocyte (H:L) ratios in two ecotypes of the garter snake Thamnophis elegans. We found that CORT and H:L ratios were not correlated within individuals, and both variables showed little or no repeatability over a period of months. CORT levels, but not H:L ratios, were higher for individuals sampled after 10min from the time of capture. However, both variables showed similar patterns of ecotypic variation, and both increased over time in gravid females maintained in captivity for four months. We suggest that CORT and H:L ratios are both useful, but disparate indices of stress in this species, and may show complex relationships to each other and to ecological and anthropogenic variables. PMID:24713520

Sparkman, Amanda M; Bronikowski, Anne M; Williams, Shelby; Parsai, Shikha; Manhart, Whitney; Palacios, Maria G



Pathology of diseases in wild desert tortoises from California.  


Twenty-four ill or dead desert tortoises (Gopherus agassizii) were received between March 1992 and July 1995 for necropsies from the Mojave and Colorado deserts of California (USA). Diseases observed in these animals included cutaneous dyskeratosis (n = 7); shell necrosis (n = 2); respiratory diseases (n = 7); urolithiasis (n = 3); and trauma (n = 5). In tortoises with cutaneous dyskeratosis the horn layer of shell was disrupted by multiple crevices and fissures and, in the most severe lesions, dermal bone showed osteoclastic resorption, remodeling, and osteopenia. In tortoises with shell necrosis, multiple foci of necrotic cell debris and heterophilic inflammation within the epidermal horn layer were subtended by necrotic dermal bone colonized by bacteria and fungi. Of the seven tortoises with respiratory disease, five were diagnosed with mycoplasmosis. The diagnosis of mycoplasmosis was based on the presence of chronic proliferative rhinitis and positive serologic tests and/or isolation of Mycoplasma sp. Chronic fungal pneumonia was diagnosed in one tortoise with respiratory disease. In the three tortoises with urolithiasis, two were discovered dead, and the live tortoise had renal and articular gout. Traumatic injuries consisted of one tortoise entombed within its burrow, one tortoise burned in a brush fire, two tortoises struck by moving vehicles, and one tortoise attacked by a predator. While the primary cause of illness could be attributed to one or two major disease processes, lesions were often found in multiple organ systems, and a variety of etiologies were responsible for morbidity and mortality. PMID:9706560

Homer, B L; Berry, K H; Brown, M B; Ellis, G; Jacobson, E R



Chickens treated with a nitric oxide inhibitor became more resistant to Plasmodium gallinaceum infection due to reduced anemia, thrombocytopenia and inflammation.  


Malaria is a serious infectious disease caused by parasites of the Plasmodium genus that affect different vertebrate hosts. Severe malaria leads to host death and involves different pathophysiological phenomena such as anemia, thrombocytopenia and inflammation. Nitric oxide (NO) is an important effector molecule in this disease, but little is known about its role in avian malaria models. Plasmodium gallinaceum-infected chickens were treated with aminoguanidine (AG), an inhibitor of inducible nitric oxide synthase, to observe the role of NO in the pathogenesis of this avian model. AG increased the survival of chickens, but also induced higher parasitemia. Treated chickens demonstrated reduced anemia and thrombocytopenia. Moreover, erythrocytes at different stages of maturation, heterophils, monocytes and thrombocytes were infected by Plasmodium gallinaceum and animals presented a generalized leucopenia. Activated leukocytes and thrombocytes with elongated double nuclei were observed in chickens with higher parasitemia; however, eosinophils were not involved in the infection. AG reduced levels of hemozoin in the spleen and liver, indicating lower inflammation. Taken together, the results suggest that AG reduced anemia, thrombocytopenia and inflammation, explaining the greater survival rate of the treated chickens. PMID:23398940

de Macchi, Barbarella Matos; Miranda, Farlen José Bebber; de Souza, Fernanda Silva; de Carvalho, Eulógio Carlos Queiroz; Albernaz, Antônio Peixoto; do Nascimento, José Luiz Martins; DaMatta, Renato Augusto



Leukocyte ultrastructure, hematological and serum biochemical profiles of ostriches (Struthio camelus).  


In the last decade ostrich farms spread throughout the world as an alternative source of investment. Although previous studies have reported hematology and biochemical values for ostriches from several regions of the world, little information is available regarding leukocyte morphology. This study reports the morphology and ultrastructure of ostrich leukocytes and hematology and biochemical values from birds raised in Brazil. Heterophils presented a lobulated nucleus, and fusiform, and acidophilic and peroxidase-negative granules. Ultrastructurally, 2 kinds of cytoplasmic granules were observed: one was large and fusiform and the other smaller with heterogeneous morphology and electrondensity; granules were peroxidase-negative. Eosinophils had a kidney-shaped eccentrically placed nucleus that was rarely lobulated and eosinophilic, round, and peroxidase-positive granules. At the ultrastructure level, 2 main kinds of granules with the same size and form but different electron density were seen; granules were peroxidase-positive. Lymphocytes and thrombocytes had the same characteristics of other avian species; monocytes presented morphological heterogeneity. Hematological and serum biochemical profiles had no sex influence and were established for ostriches raised in southeastern Brazil. These parameters will help the diagnosis of specific ostrich pathologies and serve as basic knowledge for studies in immunology and comparative avian pathology. PMID:19834079

Bonadiman, S F; Stratievsky, G C; Machado, J A; Albernaz, A P; Rabelo, G R; Damatta, R A



Cell-surface antigens of melanoma recognized by human monoclonal antibodies.  

PubMed Central

Human monoclonal antibodies (mAbs) were derived from lymph node lymphocytes and peripheral blood lymphocytes (PBL) from patients with melanoma. Four methods for generating human mAbs were compared: fusion with human [LICR-LON-HMy-2 (LICR-2)] or mouse (NS-1) cells; transformation by Epstein-Barr virus (EBV); and EBV transformation followed by NS-1 fusion. NS-1 fusion with lymph node lymphocytes resulted in a higher number of growing hybrids than LICR-2 fusion. Virtually no hybrids were obtained from NS-1 or LICR-2 fusions with PBL. EBV transformed lymphocytes from lymph node and peripheral blood with equal efficiency, and the yield of proliferating cultures for antibody screening was more than 10- to 30-fold greater than that obtained by fusion techniques. However, once antibody-producing cultures had been identified, stability and clonability of EBV-transformed cells were poorer than that of NS-1 hybrid cells. To combine the strengths of both methods, cultures of EBV-transformed cells were fused with NS-1; and hybrid clones were isolated that showed vigorous growth, clonability, and stable antibody secretion. Detailed specificity analysis of the mAbs produced by six of these clones indicated detection of a class 1 (unique) melanoma antigen, a class 3 melanoma antigen, and four ganglioside antigens (GD3, GM3, and two other, as yet uncharacterized, heterophile antigens). Images

Yamaguchi, H; Furukawa, K; Fortunato, S R; Livingston, P O; Lloyd, K O; Oettgen, H F; Old, L J



Dietary supplementation of Zingiber officinale and Zingiber zerumbet to heat-stressed broiler chickens and its effect on heat shock protein 70 expression, blood parameters and body temperature.  


The present study was conducted to assess the effects of dietary supplementation of Zingiber officinale and Zingiber zerumbet and to heat-stressed broiler chickens on heat shock protein (HSP) 70 density, plasma corticosterone concentration (CORT), heterophil to lymphocyte ratio (HLR) and body temperature. Beginning from day 28, chicks were divided into five dietary groups: (i) basal diet (control), (ii) basal diet +1%Z. zerumbet powder (ZZ1%), (iii) basal diet +2%Z. zerumbet powder (ZZ2%), (iv) basal diet +1%Z. officinale powder (ZO1%) and (v) basal diet +2%Z. officinale powder (ZO2%). From day 35-42, heat stress was induced by exposing birds to 38±1°C and 80% RH for 2 h/day. Irrespective of diet, heat challenge elevated HSP70 expression, CORT and HLR on day 42. On day 42, following heat challenge, the ZZ1% birds showed lower body temperatures than those of control, ZO1% and ZO2%. Neither CORT nor HLR was significantly affected by diet. The ZO2% and ZZ2% diets enhanced HSP70 expression when compared to the control groups. We concluded that dietary supplementation of Z. officinale and Z. zerumbet powder may induce HSP70 reaction in broiler chickens exposed to heat stress. PMID:22533311

Hasheimi, S R; Zulkifli, I; Somchit, M N; Zunita, Z; Loh, T C; Soleimani, A F; Tang, S C



Pharmacology of Cell Adhesion Molecules of the Nervous System  

PubMed Central

Cell adhesion molecules (CAMs) play a pivotal role in the development and maintenance of the nervous system under normal conditions. They also are involved in numerous pathological processes such as inflammation, degenerative disorders, and cancer, making them attractive targets for drug development. The majority of CAMs are signal transducing receptors. CAM-induced intracellular signalling is triggered via homophilic (CAM-CAM) and heterophilic (CAM - other counter-receptors) interactions, which both can be targeted pharmacologically. We here describe the progress in the CAM pharmacology focusing on cadherins and CAMs of the immunoglobulin (Ig) superfamily, such as NCAM and L1. Structural basis of CAM-mediated cell adhesion and CAM-induced signalling are outlined. Different pharmacological approaches to study functions of CAMs are presented including the use of specific antibodies, recombinant proteins, and synthetic peptides. We also discuss how unravelling of the 3D structure of CAMs provides novel pharmacological tools for dissection of CAM-induced signalling pathways and offers therapeutic opportunities for a range of neurological disorders.

Kiryushko, Darya; Bock, Elisabeth; Berezin, Vladimir



An NCAM-derived FGF-receptor agonist, the FGL-peptide, induces neurite outgrowth and neuronal survival in primary rat neurons.  


The Neural Cell Adhesion Molecule (NCAM) plays a crucial role in development of the central nervous system regulating cell migration, differentiation and synaptogenesis. NCAM mediates cell-cell adhesion through homophilic NCAM binding, subsequently resulting in activation of the fibroblast growth factor receptor (FGFR). NCAM-mediated adhesion leads to activation of various intracellular signal transduction pathways, including the Ras-mitogen activated protein kinase (MAPK) and the phosphatidylinositol-3-kinase (PI3K)-Akt pathways. A synthetic peptide derived from the second fibronectin type III module of NCAM, the FGL peptide, binds to and induces phosphorylation of FGFR without prior homophilic NCAM binding. We here present evidence that this peptide is able to mimic NCAM heterophilic binding to the FGFR by inducing neuronal differentiation as reflected by neurite outgrowth through a direct interaction with FGFR in primary cultures of three different neuronal cell types all expressing FGFR subtype 1: dopaminergic, hippocampal and cerebellar granule neurons. Moreover, we show that the FGL peptide promotes neuronal survival upon induction of cell death in the same three cell types. The effects of the FGL peptide are shown to depend on activation of FGFR and the MAPK and PI3K intracellular signalling pathways, all three kinases being necessary for the effects of FGL on neurite outgrowth and neuronal survival. PMID:15525346

Neiiendam, Johanne Louise; Køhler, Lene Boding; Christensen, Claus; Li, Shizhong; Pedersen, Martin Volmer; Ditlevsen, Dorte Kornerup; Kornum, Martin Kirkegaard; Kiselyov, Vladislav V; Berezin, Vladimir; Bock, Elisabeth



Substrate-dependent modulation of 3D spheroid morphology self-assembled in mesenchymal stem cell-endothelial progenitor cell coculture.  


The structural evolution of three-dimensional spheroids self-assembled from two different types of cells on selective biomaterials is demonstrated in this study. The two types of cells involved in the self-assembly are human mesenchymal stem cells (MSCs) and endothelial progenitor cells (EPCs). When seeded in different population ratios, they can create a variety of cellular patterns on different biomaterial substrates. When the two populations are matched in initial numbers, they are self-assembled in co-spheroids with different morphologies (i.e. randomly mixed, bumped, or concentric spheroids). The morphologies are influenced by the specific cell-substrate interaction possibly through integrin signaling, as well as a substrate-dependent regulation of heterophilic cell-cell interaction possibly through Notch signaling. In particular, the self-assembled core-shell concentric spheroids from adipose-derived MSCs and EPCs show a greater angiogenic effect in vitro. This study reveals the possibility to modulate the self-assembled morphology as well as the effect of cocultured cells by changing the cell culture substratum. PMID:24909102

Hsu, Shan-Hui; Ho, Tung-Tso; Huang, Nien-Chi; Yao, Chao-Ling; Peng, Luen-Hau; Dai, Niann-Tzyy



Hematology and blood biochemistry of captive mugger crocodiles (Crocodylus palustris).  


Comprehensive hematologic and biochemical analyses were conducted on blood from 23 male and 31 female clinically stable captive mugger crocodiles (Crocodylus palustris). Erythrocyte mean corpuscular volume (MCV), potassium, cholesterol, and calcium concentrations were significantly greater in juvenile males than in juvenile females, but no significant differences were determined between parameters of subadult males and subadult females. The mean WBC count and mean heterophil count were significantly higher in adult males than in adult females. Mean uric acid concentration was significantly greater in adult females than in males. Mean erythrocyte count was significantly higher in adults than in juveniles. Adult mean WBC and lymphocyte counts were significantly lower than those of both juveniles and subadults. Subadults had significantly lower mean eosinophil counts than both adults and juveniles. Subadults had significantly lower mean alkaline phosphatase activities than juveniles, whereas the adults had significantly lower aspartate aminotransferase and alanine aminotransferase activities than other groups. Lactate dehydrogenase activities were significantly lower for subadults than for juveniles and adults. Cholesterol concentrations were significantly higher for subadults and juveniles compared with adults. Triglyceride concentration was significantly lower for subadults and highest for juveniles. Glucose concentrations were significantly higher for adults. Blood urea nitrogen was significantly lower for subadults than for both adults and juveniles. Uric acid concentrations were significantly higher for juveniles than for the subadults and adults. The subadult animals also had a significantly lower potassium concentration. The results obtained were then compared with known values for other crocodilian species. PMID:11237141

Stacy, B A; Whitaker, N



Acetylcholinesterase-transgenic mice display embryonic modulations in spinal cord choline acetyltransferase and neurexin I? gene expression followed by late-onset neuromotor deterioration  

PubMed Central

To explore the possibility that overproduction of neuronal acetylcholinesterase (AChE) confers changes in both cholinergic and morphogenic intercellular interactions, we studied developmental responses to neuronal AChE overexpression in motoneurons and neuromuscular junctions of AChE-transgenic mice. Perikarya of spinal cord motoneurons were consistently enlarged from embryonic through adult stages in AChE-transgenic mice. Atypical motoneuron development was accompanied by premature enhancement in the embryonic spinal cord expression of choline acetyltransferase mRNA, encoding the acetylcholine-synthesizing enzyme choline acetyltransferase. In contrast, the mRNA encoding for neurexin-I?, the heterophilic ligand of the AChE-homologous neuronal cell surface protein neuroligin, was drastically lower in embryonic transgenic spinal cord than in controls. Postnatal cessation of these dual transcriptional responses was followed by late-onset deterioration in neuromotor performance that was associated with gross aberrations in neuromuscular ultrastructure and with pronounced amyotrophy. These findings demonstrate embryonic feedback mechanisms to neuronal AChE overexpression that are attributable to both cholinergic and cell–cell interaction pathways, suggesting that embryonic neurexin I? expression is concerted in vivo with AChE levels and indicating that postnatal changes in neuronal AChE-associated proteins may be involved in late-onset neuromotor pathologies.

Andres, Christian; Beeri, Rachel; Friedman, Alon; Lev-Lehman, Efrat; Henis, Sivan; Timberg, Rina; Shani, Moshe; Soreq, Hermona



Hematology and blood chemistry values in free-living imperial cormorants (Phalacrocorax atriceps).  


As part of an on-going, long-term study on the reproductive ecology and health status of imperial cormorants (Phalacrocorax atriceps), blood samples were collected to establish baseline values for hematologic parameters (hematocrit, red and white blood cell counts, leukocyte profile, heterophil:lymphocyte ratio, total solids) and serum chemistries (glucose, uric acid, urea, total protein, triglycerides, cholesterol, albumin:globulin ratio, alkaline phosphatase, lactate dehydrogenase, creatine phosphokinase, aspartate aminotransferase, alanine aminotransferase, calcium, phosphorus). One hundred and eighty-four male adults from the Punta Le6n breeding colony in Patagonia Argentina were captured during the chick-rearing period of four breeding seasons, 2004 (n = 48), 2005 (n = 29), 2010 (n = 43), and 2011 (n = 64). All birds appeared to be in good body condition and no abnormalities were noted during physical examination. In general, values for the parameters reported in this study were similar to those previously described for other cormorant species. Significant interannual differences were observed in most health parameters analyzed. This study defines baseline health parameters for imperial cormorants and, coupled with previous reports on pathogen exposure, contributes to our knowledge of the overall health status of the species. PMID:24597115

Gallo, Luciana; Quintana, Flavio; Svagelj, Walter S; Uhart, Marcela



Effects of ascorbic acid on stress and disease in chickens.  


White leghorn chickens were given feed containing 100 mg of ascorbic acid (AA)/kg. One day later, treated chickens and a similar group of unmedicated control chickens were chilled for 1 hour at 6 C, exposed to an unusual sound, fasted, or subjected to rough handling. Heterophil:lymphocyte (H:L) ratios were determined one day later. The AA-treated birds had significantly lower H:L ratios than untreated controls. Chickens that received a diet containing AA had lower H:L ratios than controls (0.86 vs. 1.65) following administration of adrenocorticotropic hormone. Chickens fed a diet containing AA showed increased resistance to a combined Newcastle disease virus-Mycoplasma gallisepticum infection and to a secondary Escherichia coli infection, as well as to a primary E. coli challenge infection. The effects of AA and an antibacterial drug (furaltadone) were additive. In all experiments, the optimum dose of AA was 100 mg/kg of feed. There was a negative correlation between AA level in the diet and feed efficiency. PMID:1417599

Gross, W B



Elucidating the principles of the molecular organization of heteropolymeric tight junction strands.  


Paracellular barrier properties of tissues are mainly determined by the composition of claudin heteropolymers. To analyze the molecular organization of tight junctions (TJ), we investigated the ability of claudins (Cld) to form homo- and heteromers. Cld1, -2, -3, -5, and -12 expressed in cerebral barriers were investigated. TJ-strands were reconstituted by claudin-transfection of HEK293-cells. cis-Interactions and/or spatial proximity were analyzed by fluorescence resonance energy transfer inside and outside of strands and ranked: Cld5/Cld5 > Cld5/Cld1 > Cld3/Cld1 > Cld3/Cld3 > Cld3/Cld5, no Cld3/Cld2. Classic Cld1, -3, and -5 but not non-classic Cld12 showed homophilic trans-interaction. Freeze-fracture electron microscopy revealed that, in contrast to classic claudins, YFP-tagged Cld12 does not form homopolymers. Heterophilic trans-interactions were analyzed in cocultures of differently monotransfected cells. trans-Interaction of Cld3/Cld5 was less pronounced than that of Cld3/Cld1, Cld5/Cld1, Cld5/Cld5 or Cld3/Cld3. The barrier function of reconstituted TJ-strands was demonstrated by a novel imaging assay. A model of the molecular organization of TJ was generated. PMID:21533891

Piontek, Jörg; Fritzsche, Susanne; Cording, Jimmi; Richter, Sandra; Hartwig, Jens; Walter, Maria; Yu, Dan; Turner, Jerrold R; Gehring, Claudia; Rahn, Hans-Peter; Wolburg, Hartwig; Blasig, Ingolf E



Participation of the second extracellular loop of claudin-5 in paracellular tightening against ions, small and large molecules.  


Tight junctions control paracellular permeability. Here, we analyzed the impact of residues in the second extracellular loop (ECL2) of mouse claudin-5 on paracellular permeability. Stable expression of claudin-5(wild type) in MDCK-II cells-but not that of mutants R145A, Y148A, Y158A or E159Q-increased transepithelial electrical resistance and decreased fluorescein permeation. Expression of claudin-5(Y148A), (Y158A) or (E159Q) enhanced permeability of FITC-dextran(10 kDa), which was unchanged in cells expressing claudin-5(wild type) or claudin-5(R145A). In contrast, targeting to tight junctions, strand morphology and tight junction assembly were unchanged. It is concluded that R145 is unessential for trans-interaction of claudin-5, but necessary for tightening against small solutes and ions. The highly conserved residues Y148, Y158 and E159 in ECL2 of claudin-5 contribute to homo- and/or heterophilic trans-interaction between classic claudins and thereby tighten the paracellular space against ions, small and large molecules. These results provide novel insights into the molecular function of tight junctions. PMID:20333434

Piehl, Christian; Piontek, Jörg; Cording, Jimmi; Wolburg, Hartwig; Blasig, Ingolf E



Hematological and plasma biochemical reference ranges of Alaskan seabirds: Their ecological significance and clinical importance  

USGS Publications Warehouse

Blood was analyzed from 151 pelagic marine birds to establish reference ranges for hematological and plasma biochemical parameters from healthy, wild populations of Pacific seabirds. Of the 13 species examined, 9 were from the Family Alcidae (N = 122 individuals) and the remainder (N = 29) from the Families Phalacrocoracidae, Laridae, and Procellariidae. Three of 8 hematological parameters (total white blood cell count, lymphocyte count and eosinophil count) differed significantly among species, as did 9 of 13 plasma biochemical parameters (alkaline phosphatase, aspartate aminotransferase, creatine kinase, cholesterol, glucose, lactate dehydrogenase, total bilirubin, total protein and field total protein). There were no differences among species for packed cell volume, buffy coat, cell counts of heterophils, monoqtes and basophils, or for concentrations of alanine aminotransferase, triglycerides, uric acid and calcium. Plasma calcium concentration, triglyceride levels and field total protein varied significantly between sexes, with females having higher mean concentrations of all 3 parameters. However, no significant relationships between measures of breeding condition (brood patch size, subcutaneous and mesenteric fat deposits, or ovarian follicle size and ovary weight) and calcium or alkaline phosphatase concentrations in female birds could be identified. Alanine aminotransferase and uric acid were the only analytes which did not differ significantly between species or sexes.

Newman, S. H.; Piatt, J. F.; White, J.



Infectious Mononucleosis and Mononucleosis Syndromes  

PubMed Central

Infectious mononucleosis (IM) and cytomegalovirus (CMV) mononucleosis are caused by a primary infection with related viruses, Epstein-Barr virus (EBV) and CMV. Despite the similarity of clinical manifestations, basic differences exist: (1) The heterophil antibody (HA) response is absent in CMV mononucleosis, whereas it is present in IM. (2) In IM atypical lymphocytosis reflects proliferation of B cells early and of T cells later in the disease course; in CMV mononucleosis the situation appears complex. (3) In blood, EBV is restricted to B lymphocytes, whereas CMV is found in polymorphonuclear and mononuclear leukocytes. (4) Complications of CMV mononucleosis such as hepatitis and pneumonitis may be due to virus cytopathic effect in target organs. Prominent tonsillopharyngitis with adenopathy, and visceral complications of IM are related to lymphoproliferation which is self-limited except in males with a rare familial defect in defense against EBV. Immune complex-mediated pathology may occur in both diseases. (5) CMV is frequently transmitted to a fetus in utero or to an infant during or after birth, and this occasionally leads to severe cytomegalic inclusion disease; vertical transmission of EBV appears to be exceptional. (6) Secondary EBV infections are associated with certain malignancies whereas such an association has not been recognized in the case of CMV. Toxoplasma gondii is another cause of HA-negative mononucleosis. Its complications in the heart, in skeletal muscle and in the central nervous system are related to direct invasion by the parasite. Cellular immunity plays an important role in defense against all three agents.

Fiala, Milan; Heiner, Douglas C.; Turner, Jerrold A.; Rosenbloom, Barry; Guze, Lucien B.



In vivo studies of Gallibacterium anatis infection in chickens.  


The aim of the present study was to investigate the pathology in normal or immunosuppressed chickens followed intravenous or intraperitoneal inoculation with a well-characterized strain of Gallibacterium anatis. Two groups of 30 15-week-old commercial brown laying chickens were used, having been screened and found negative for Gallibacterium organisms. One group was treated with 5-fluorouracil to promote heterophil depletion, while the other was saline treated. Ten days later 15 chickens from each group were inoculated either intravenously or intraperitoneally with 3.3 x 10(7) colony-forming units of G. anatis strain 12656-12. Subsets of chickens were sacrificed at 3, 12 or 24 h post-infection and examined for lesions. Livers and spleens were examined by culture and by fluorescent in situ hybridization. Intravenously infected birds showed severe septicaemic lesions in both the normal and immunosuppressed birds. Mortality was recorded only in the latter, with an overall rate of 73%. The intraperitoneally infected chickens of normal immune status showed various degrees of localized purulent peritonitis at the inoculation site, but in the immunosuppressed birds the entire peritoneum tended to be involved along with the abdominal organs. This was similar to previous descriptions of natural infections and may represent a useful infection model for detailed analysis of Gallibacterium virulence factors and pathogenesis. PMID:15276980

Bojesen, Anders Miki; Nielsen, Ole Lerberg; Christensen, Jens Peter; Bisgaard, Magne



Metal accumulation and evaluation of effects in a freshwater turtle.  


A variety of contaminants have been detected in aquatic and terrestrial environments around the Paducah Gaseous Diffusion Plant (PGDP), Kentucky. The presence of these contaminants at the PGDP may pose a risk to biota, yet little is known about the bioaccumulation of contaminants and associated effects in wildlife, especially in aquatic turtles. The current study was initiated to evaluate: (1) the accumulation of heavy metals (Cd, Cr, Cu, Pb, and Hg) in aquatic ecosystems associated with the PGDP using red-eared slider turtle (Trachemys scripta elegans) as biomonitors; (2) maternal transfer of heavy metals; and (3) potential hematological and immunological effects resulting from metal accumulation. A total of 26 turtles were collected from 7 ponds located south, adjacent, and north of the PGDP. Liver Cu concentrations were significantly different among ponds and Cu concentrations in eggs were positively correlated with female Cu concentrations in kidney. The concentrations of heavy metals measured in turtle tissues and eggs were low and, based on previous studies of reptiles and established avian threshold levels of heavy metals, did not appear to have adverse effects on aquatic turtles inhabiting ponds near the PGDP. However, total white blood cell counts, heterophil to lymphocyte ratio, and phytohemagglutinin stimulation index were correlated with metal concentrations. Because other factors may affect the hematological and immunological indices, further investigation is needed to determine if these effects are associated with metal exposure, other contaminants, or disease. PMID:21688058

Yu, Shuangying; Halbrook, Richard S; Sparling, Donald W; Colombo, Robert



Accumulation of polychlorinated biphenyls (PCBs) and evaluation of hematological and immunological effects of PCB exposure on turtles.  


Concentrations of total polychlorinated biphenyls (PCBs), Aroclor 1260, and 26 congeners were measured in liver, fat, and eggs of red-eared slider turtles (Trachemys scripta elegans) collected from ponds near or on the Paducah Gaseous Diffusion Plant (PGDP), Kentucky, USA. Concentrations of total PCBs (wet mass) ranged from 0.002 to 0.480 mg/kg, 0.028 to 0.839 mg/kg, and 0.001 to 0.011 mg/kg in liver, fat, and eggs, respectively. Concentrations of Arochlor 1260 did not exceed 0.430, 0.419, and 0.007 mg/kg in liver, fat, and eggs, respectively. Exposure to PCBs in red-eared sliders collected from the PGDP is characterized by low concentrations of moderately chlorinated mono-ortho and di-ortho congeners (PCB 153, 180, and 118). Although PCB concentrations measured in the current study were low, chronic exposure to PCBs may have altered hematology and immunity of the turtles examined. Total white blood cell count and number of heterophils were negatively correlated with concentrations of total PCBs and Arochlor 1260, respectively. However, disease and other contaminants in the study area may influence the results. Because little is known regarding the influence of PCBs on hematology and immune function in turtles, additional study is needed to better evaluate results observed in the current study. PMID:22430883

Yu, Shuangying; Halbrook, Richard S; Sparling, Donald W



Membrane-impermeable Cross-linking Provides Evidence for Homophilic, Isoform-specific Binding of Desmosomal Cadherins in Epithelial Cells*?  

PubMed Central

Desmosomes and adherens junctions are cadherin-based protein complexes responsible for cell-cell adhesion of epithelial cells. Type 1 cadherins of adherens junctions show specific homophilic adhesion that plays a major role in developmental tissue segregation. The desmosomal cadherins, desmocollin and desmoglein, occur as several different isoforms with overlapping expression in some tissues where different isoforms are located in the same desmosomes. Although adhesive binding of desmosomal cadherins has been investigated in a variety of ways, their interaction in desmosome-forming epithelial cells has not been studied. Here, using extracellular homobifunctional cross-linking, we provide evidence for homophilic and isoform-specific binding between the Dsc2, Dsc3, Dsg2, and Dsg3 isoforms in HaCaT keratinocytes and show that it represents trans interaction. Furthermore, the cross-linked adducts are present in the detergent-insoluble fraction, and electron microscopy shows that extracellular cross-linking probably occurs in desmosomes. We found no evidence for either heterophilic or cis interaction, but neither can be completely excluded by our data. Mutation of amino acid residues Trp-2 and Ala-80 that are important for trans interaction in classical cadherin adhesive binding abolished Dsc2 binding, indicating that these residues are also involved in desmosomal adhesion. These interactions of desmosomal cadherins may be of key importance for their ordered arrangement within desmosomes that we believe is essential for desmosomal adhesive strength and the maintenance of tissue integrity.

Nie, Zhuxiang; Merritt, Anita; Rouhi-Parkouhi, Mansour; Tabernero, Lydia; Garrod, David



Mating type-specific cell-cell recognition of Saccharomyces cerevisiae: cell wall attachment and active sites of a- and alpha-agglutinin.  

PubMed Central

Mating type-specific agglutination of Saccharomyces cerevisiae a and alpha cells depends on the heterophilic interaction of two cell surface glycoproteins, the gene products of AG alpha 1 and AGA2. Evidence is presented with immunogold labelling that the alpha-agglutinin is part of the outer fimbrial cell wall coat. The a-agglutinin is bound via two S-S bridges (Cys7 and Cys50) to a cell wall component, most probably the gene product of AGA1. His273 of alpha-agglutinin has previously been shown to be essential for a- and alpha-agglutinin interaction and a model based on two opposing ion-pairs had been proposed. By site-directed mutagenesis this possibility has now been excluded. With the help of various peptides, either chemically synthesized, obtained by proteolysis of intact glycosylated a-agglutinin or prepared from a fusion protein expressed in Escherichia coli, the biologically active region of a-agglutinin was located at the C-terminus of the molecule. A peptide consisting of the C-terminal 10 amino acids (GSPIN-TQYVF) was active in nanomolar concentrations. Saccharide moieties, therefore, are not essential for the mating type-specific cell-cell interaction; glycosylated peptides are, however, four to five times more active than non-glycosylated ones. Comparisons of the recognition sequences of the S. cerevisiae agglutinins with that of the Dictyostelium contact site A glycoprotein (gp80), as well as with those of the various families of cell adhesion molecules of higher eucaryotes, have been made and are discussed. Images

Cappellaro, C; Baldermann, C; Rachel, R; Tanner, W



Mating type-specific cell-cell recognition of Saccharomyces cerevisiae: cell wall attachment and active sites of a- and alpha-agglutinin.  


Mating type-specific agglutination of Saccharomyces cerevisiae a and alpha cells depends on the heterophilic interaction of two cell surface glycoproteins, the gene products of AG alpha 1 and AGA2. Evidence is presented with immunogold labelling that the alpha-agglutinin is part of the outer fimbrial cell wall coat. The a-agglutinin is bound via two S-S bridges (Cys7 and Cys50) to a cell wall component, most probably the gene product of AGA1. His273 of alpha-agglutinin has previously been shown to be essential for a- and alpha-agglutinin interaction and a model based on two opposing ion-pairs had been proposed. By site-directed mutagenesis this possibility has now been excluded. With the help of various peptides, either chemically synthesized, obtained by proteolysis of intact glycosylated a-agglutinin or prepared from a fusion protein expressed in Escherichia coli, the biologically active region of a-agglutinin was located at the C-terminus of the molecule. A peptide consisting of the C-terminal 10 amino acids (GSPIN-TQYVF) was active in nanomolar concentrations. Saccharide moieties, therefore, are not essential for the mating type-specific cell-cell interaction; glycosylated peptides are, however, four to five times more active than non-glycosylated ones. Comparisons of the recognition sequences of the S. cerevisiae agglutinins with that of the Dictyostelium contact site A glycoprotein (gp80), as well as with those of the various families of cell adhesion molecules of higher eucaryotes, have been made and are discussed. PMID:7957044

Cappellaro, C; Baldermann, C; Rachel, R; Tanner, W



Evaluation of an Enzyme Immunoassay for Detection of Immunoglobulin M Antibodies to West Nile Virus and the Importance of Background Subtraction in Detecting Nonspecific Reactivity?  

PubMed Central

Since the introduction of West Nile virus (WNV) in the United States in 1999, several assays have become commercially available to detect antibodies against WNV. Capture enzyme-linked immunosorbent assays (ELISAs) for the detection of WNV-specific immunoglobulin M (IgM) have been approved by the Food and Drug Administration for clinical testing and are available from Focus Diagnostics and PanBio, Inc. The Focus Diagnostics IgM capture ELISA utilizes a background subtraction protocol in order to detect nonspecific reactivity due to rheumatoid factor, heterophile antibodies, or other interfering substances. A background subtraction procedure is not currently recommended for the PanBio IgM capture ELISA. In previous experiments, we determined the agreement, sensitivity, and specificity of the PanBio first-generation IgM capture ELISA compared to an immunofluorescence assay and the Centers for Disease Control and Prevention's IgM capture ELISA. The PanBio assay has since been reformulated to improve the specificity of the assay. We evaluated the reformulated PanBio assay with and without an antigen subtraction procedure and compared the results to the Focus IgM capture ELISA. Agreement, sensitivity, and specificity of the PanBio assay were, respectively, 85%, 95%, and 76% without the subtraction protocol and 94%, 95%, and 93% with the subtraction protocol. In general, when the subtraction protocol was applied to the PanBio IgM capture ELISA, there was a reduction in some, but not all, false-positive results. We suggest that all WNV IgM assays be standardized with a procedure such as background subtraction to eliminate nonspecific reactivity that may cause false-positive results.

Rawlins, Mindy L.; Swenson, Erica M.; Hill, Harry R.; Litwin, Christine M.



Hematologic and biochemical reference ranges for captive California condors (Gymnogyps californianus).  


To provide proper medical evaluation and care for the endangered California condor (Gymnogyps californianus), veterinarians need accurate hematologic and biochemical reference ranges. A retrospective study of blood samples from captive California condors housed at the San Diego Wild Animal Park assessed the samples by sex and age of condor to determine serum biochemical and hematologic reference ranges, including lead and zinc levels. Condors were grouped by age as follows: group 1 included birds less than 30 days of age; group 2 included birds between 30 days and 6 mo of age; group 3 included birds between 6 mo and 5 yr of age; group 4 included all birds greater than 5 yr of age. Significant differences between sexes included higher chloride, cholesterol, and total plasma protein concentration in males as compared to females (P < 0.05). Significant differences between age groups were identified in glucose, potassium, phosphorus, calcium, albumin, total plasma protein, globulin, cholesterol, bile acid, and zinc concentrations, as well as aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase, and creatine phosphokinase activities (P < 0.05). Additionally, significant differences between age groups were noted in white blood cell count, hematocrit, heterophils, lymphocytes, and eosinophils (P < 0.05). A steady increase in glucose and a decrease in alkaline phosphatase and lactate dehydrogenase activities, as well as cholesterol, bile acid, calcium, and phosphorus concentrations, were correlated with age (P < 0.05). Following application of statistical analysis, condors less than 6 mo of age were identified as unique compared to older cohorts; therefore, two reference ranges are proposed by calculating a 90% confidence interval. Reference ranges obtained from other published avian data, including those for psittaciformes, ratites, galliformes, anseriformes, and raptors, were similar to condors in this study. PMID:17312714

Dujowich, Mauricio; Mazet, Jonna K; Zuba, Jeffery R



Human lung mast cells adhere to human airway smooth muscle, in part, via tumor suppressor in lung cancer-1.  


Mast cells infiltrate the airway smooth muscle (ASM) of patients with asthma, an event which is likely to be a key factor in the development of this disease. Adhesion is a fundamental mechanism facilitating cellular cross-talk. We have examined whether human lung mast cells (HLMC) and ASM adhere, and have also examined the mechanism involved. Primary cultures of HLMC and confluent human ASM were cocultured for 30 min, then nonadherent HLMC were removed by centrifugation. HLMC adhered avidly to ASM monolayers (mean +/- SEM adhesion 43.2 +/- 1.2%, n = 41). Adhesion was increased to 58.8 +/- 2.7% by 1 mM Mn2+ (p = 0.015), and was reduced by EDTA and EGTA to 20.5 +/- 1.5% and 21.0 +/- 1.3%, respectively (p < 0.0001). Adhesion-blocking Abs for ICAM-1, VCAM-1, CD18, and the alpha4 and beta1 integrins had no effect on HLMC adhesion. HLMC expressed tumor suppressor in lung cancer-1 (TSLC-1) and blocking this reduced adhesion from 38.5 +/- 4.8% to 28.3 +/- 3.7% (p = 0.004, n = 7). ASM did not express TSLC-1, indicating that TSLC-1 acts as a heterophilic adhesion molecule. In summary, HLMC adhere avidly to ASM in part via TSLC-1 and in part via an as-yet-undefined Ca2+-dependent pathway. This supports the hypothesis that adhesion is important in the recruitment and retention of HLMC by the ASM in asthma, and for the functional interaction of these cells. PMID:16394014

Yang, Weidong; Kaur, Davinder; Okayama, Yoshimichi; Ito, Akihiko; Wardlaw, Andrew J; Brightling, Christopher E; Bradding, Peter



A note on protein expression changes in chicken breast muscle in response to time in transit before slaughtering  

PubMed Central

Aims of the research were to devise a proteome map of the chicken Pectoralis superficialis muscle, as resolved by two-dimensional gel electrophoresis, and to characterize protein expression changes in the soluble protein fraction in commercial conditions due to age and to time in transit before slaughtering. Broilers were reared under commercial conditions until they reached a mean 1.8 kg and 36 d, or 2.6 kg and 46 d of age. Transport to the slaughterhouse took 90 or 220 minutes. Transport-induced stress was assessed from blood metabolites and leukocyte cell counts, revealing significant changes in albumin, glucose and triglyceride concentrations, in heterophils and leukocyte counts for chickens in transit for longer, and in glucose depending mainly on age. The sarcoplasmic protein fractions were extracted from a total of 39 breast muscle samples, collected 15 min post mortem, for analysis by two-dimensional electrophoresis. Image and statistical analyses enabled us to study the qualitative and quantitative differences between the samples. Twelve up- or down-regulated protein spots were detected (P?



Differential Effects of Human L1CAM Mutations on Complementing Guidance and Synaptic Defects in Drosophila melanogaster  

PubMed Central

A large number of different pathological L1CAM mutations have been identified that result in a broad spectrum of neurological and non-neurological phenotypes. While many of these mutations have been characterized for their effects on homophilic and heterophilic interactions, as well as expression levels in vitro, there are only few studies on their biological consequences in vivo. The single L1-type CAM gene in Drosophila, neuroglian (nrg), has distinct functions during axon guidance and synapse formation and the phenotypes of nrg mutants can be rescued by the expression of human L1CAM. We previously showed that the highly conserved intracellular FIGQY Ankyrin-binding motif is required for L1CAM-mediated synapse formation, but not for neurite outgrowth or axon guidance of the Drosophila giant fiber (GF) neuron. Here, we use the GF as a model neuron to characterize the pathogenic L120V, Y1070C, C264Y, H210Q, E309K and R184Q extracellular L1CAM missense mutations and a L1CAM protein with a disrupted ezrin–moesin–radixin (ERM) binding site to investigate the signaling requirements for neuronal development. We report that different L1CAM mutations have distinct effects on axon guidance and synapse formation. Furthermore, L1CAM homophilic binding and signaling via the ERM motif is essential for axon guidance in Drosophila. In addition, the human pathological H210Q, R184Q and Y1070C, but not the E309K and L120V L1CAM mutations affect outside-in signaling via the FIGQY Ankyrin binding domain which is required for synapse formation. Thus, the pathological phenotypes observed in humans are likely to be caused by the disruption of signaling required for both, guidance and synaptogenesis.

Kudumala, Sirisha; Freund, Julie; Hortsch, Michael; Godenschwege, Tanja A.



Protein H--a bacterial surface protein with affinity for both immunoglobulin and fibronectin type III domains.  

PubMed Central

Several bacterial species express surface proteins with affinity for the constant region (Fc) of immunoglobulin (Ig) G. The biological consequences of the interaction with IgG are poorly understood but it has been demonstrated that genes encoding different IgG Fc-binding proteins have undergone convergent evolution, suggesting that these surface molecules are connected with essential microbial functions. One of the molecules, protein H, is present in some strains of Streptococcus pyogenes, the most significant streptococcal species in clinical medicine. In contrast to other Ig-binding bacterial proteins tested, protein H was found to interact also with the neural cell adhesion molecule (N-CAM), a eukaryotic cell surface glycoprotein mediating homo- and heterophilic cell-cell interactions. The affinity for the interaction between protein H and N-CAM was 1.6 x 10(8)/M and the binding site on protein H was mapped to the NH2-terminal 80 amino acid residues. N-CAM and IgG are both members of the Ig superfamily and analogous to N-CAM, IgG binds to the NH2-terminal part of protein H. However, the binding sites for the two proteins were found to be separate, an unexpected result which was explained by the observation that the fibronectin type III (FNIII) domains and not the Ig-like domains of N-CAM are responsible for the interaction with protein H. Thus, the binding of N-CAM to protein H was blocked with fibronectin but not with IgG. Moreover, apart from fibronectin itself and N-CAM, fragments of fibronectin and the matrix protein cytotactin/tenascin containing FNIII domains also showed affinity for protein H.(ABSTRACT TRUNCATED AT 250 WORDS) Images

Frick, I M; Crossin, K L; Edelman, G M; Bjorck, L



N-Glycosylation at the SynCAM (Synaptic Cell Adhesion Molecule) Immunoglobulin Interface Modulates Synaptic Adhesion*  

PubMed Central

Select adhesion molecules connect pre- and postsynaptic membranes and organize developing synapses. The regulation of these trans-synaptic interactions is an important neurobiological question. We have previously shown that the synaptic cell adhesion molecules (SynCAMs) 1 and 2 engage in homo- and heterophilic interactions and bridge the synaptic cleft to induce presynaptic terminals. Here, we demonstrate that site-specific N-glycosylation impacts the structure and function of adhesive SynCAM interactions. Through crystallographic analysis of SynCAM 2, we identified within the adhesive interface of its Ig1 domain an N-glycan on residue Asn60. Structural modeling of the corresponding SynCAM 1 Ig1 domain indicates that its glycosylation sites Asn70/Asn104 flank the binding interface of this domain. Mass spectrometric and mutational studies confirm and characterize the modification of these three sites. These site-specific N-glycans affect SynCAM adhesion yet act in a differential manner. Although glycosylation of SynCAM 2 at Asn60 reduces adhesion, N-glycans at Asn70/Asn104 of SynCAM 1 increase its interactions. The modification of SynCAM 1 with sialic acids contributes to the glycan-dependent strengthening of its binding. Functionally, N-glycosylation promotes the trans-synaptic interactions of SynCAM 1 and is required for synapse induction. These results demonstrate that N-glycosylation of SynCAM proteins differentially affects their binding interface and implicate post-translational modification as a mechanism to regulate trans-synaptic adhesion.

Fogel, Adam I.; Li, Yue; Giza, Joanna; Wang, Qing; Lam, TuKiet T.; Modis, Yorgo; Biederer, Thomas



Evidence for a human-specific mechanism for diet and antibody-mediated inflammation in carcinoma progression.  


Patients with cancer have circulating heterophile antibodies that agglutinate animal red cells via recognition of the mammalian cell surface sialic acid N-glycolylneuraminic acid (Neu5Gc), which was long considered an oncofetal antigen in humans. However, humans are genetically deficient in Neu5Gc production and instead metabolically accumulate Neu5Gc from dietary sources, particularly red meats and milk products. Moreover, mice with a human-like defect showed no alternate pathway for Neu5Gc synthesis and even normal humans express anti-Neu5Gc antibodies. We show here that human tumors accumulate Neu5Gc that is covalently attached to multiple classes of glycans. The paradox of human tumor Neu5Gc accumulation in the face of circulating anti-Neu5Gc antibodies was hypothesized to be due to facilitation of tumor progression by the resulting low-grade chronic inflammation. Indeed, murine tumors expressing human-like levels of Neu5Gc show accelerated growth in syngeneic mice with a human-like Neu5Gc deficiency, coincident with the induction of anti-Neu5Gc antibodies and increased infiltration of inflammatory cells. Transfer of polyclonal monospecific syngeneic mouse anti-Neu5Gc serum also enhanced growth of transplanted syngeneic tumors bearing human-like levels of Neu5Gc, with tumors showing evidence for antibody deposition, enhanced angiogenesis and chronic inflammation. These effects were suppressed by a cyclooxygenase-2 inhibitor, a drug type known to reduce human carcinoma risk. Finally, affinity-purified human anti-Neu5Gc antibodies also accelerate growth of Neu5Gc-containing tumors in Neu5Gc-deficient mice. Taken together, the data suggest that the human propensity to develop diet-related carcinomas is contributed to by local chronic inflammation, resulting from interaction of metabolically-accumulated dietary Neu5Gc with circulating anti-Neu5Gc antibodies. PMID:19017806

Hedlund, Maria; Padler-Karavani, Vered; Varki, Nissi M; Varki, Ajit



Immune response to an endotoxin challenge involves multiple immune parameters and is consistent among the annual-cycle stages of a free-living temperate zone bird.  


Trade-offs between immune function and other physiological and behavioural processes are central in ecoimmunology, but one important problem is how to distinguish a reallocation of resources away from the immune system from a reallocation or redistribution within the immune system. While variation in baseline values of individual immune parameters is well established, studies in wild animals on multiple parameters during an immune response are lacking. It also remains to be tested whether and how immune responses correlate with baseline values that vary, for example, over the course of an annual cycle. We studied immunological responses to an endotoxin challenge in skylarks (Alauda arvensis), a partial migrant bird breeding in temperate zones. We compared birds injected with the endotoxin LPS with un-injected controls, characterizing immunological responses with leukocyte profiles, titres of lytic enzymes and natural antibodies, and concentrations of haptoglobin and heat shock proteins. We did this in five annual-cycle stages to test whether the response varied throughout the year. The endotoxin challenge affected six of 10 measured parameters. Lysis titres and proportions of heterophils increased; haptoglobin concentrations and proportions of lymphocytes, basophils and eosinophils decreased. The variable effects on different immune components demonstrate the complexity of an immune response. We found no evidence that the response differed between annual-cycle stages. The response was independent of baseline measures taken directly upon capture in the field, indicating that birds were facing no immunological ceiling when mounting an immune response. Values of five parameters collected under field conditions were significantly related to values taken under standardized laboratory conditions. We conclude that multiple parts of the immune system are modulated during an immunological response and that responses are not re-organized throughout the annual cycle. PMID:23531817

Hegemann, Arne; Matson, Kevin D; Versteegh, Maaike A; Villegas, Auxiliadora; Tieleman, B Irene



Cytokines and chemokines in postovulatory follicle regression of domestic chicken (Gallus gallus domesticus).  


The mechanism of postovulatory follicle (POF) regression in birds is still poorly understood. In the current study, expression of IL-1beta, IL-6, GM-CSF, IFN-gamma, IL-2, IL-4, IL-13, chCXCLi2, chCCLi2, chCCLi4, chCCLi7, IL-10 and TGF-beta2 mRNAs was estimated in regressing POF by semi-quantitative RT-PCR. In addition, the changes in immune cell population, histological and apoptotic changes were also studied in regressing POF. The expression of cytokines (IL-1beta, IL-6, IL-10 and TGF-beta2) and chemokines (chCXCLi2, chCCLi2, chCCLi4 and chCCLi7) was upregulated in POFs, suggesting a role for these molecules in tissue regression. The histological findings suggested a significant infiltration of immune cells, especially heterophils, lymphocytes and macrophages, into the regressing POF. The flow cytometry analysis of lymphocyte subpopulations revealed that CD3(+), CD4(+), CD8(+) and Bu-1(+) lymphocytes were significantly increased during this regression. The significant up-regulation of chemokines might have attracted the immune cells during POF regression. The percentage of apoptotic cells was significantly increased during the regression of POF. The up-regulation of IL-1beta, IL-6, IL-10 and TGF-beta2 and down-regulation of GM-CSF might have induced apoptosis during the POF regression. However, expression of IFN-gamma, IL-2, IL-4 and IL-13 was not significantly altered during POF regression. In conclusion, cytokines appear to play an important role in the regression of POF in chicken. Furthermore, the regression of chicken POF seems to be an inflammatory event similar to luteolysis of the mammalian corpus luteum. PMID:17692913

Sundaresan, N R; Saxena, V K; Sastry, K V H; Nagarajan, K; Jain, Preeti; Singh, Rani; Anish, D; Ravindra, P V; Saxena, M; Ahmed, K A



[Encephalitis due to the Epstein-Barr virus: a description of a clinical case and review of the literature].  


INTRODUCTION. Infection by the Epstein-Barr virus (EBV) -either as a primary infection, a reactivation or an active chronic infection- can give rise to several clinical forms of involvement of the central nervous system. We report a case of encephalitis due to EBV produced by viral reactivation in an immunocompetent patient which initially mimicked, from the clinical and electroencephalographic point of view, encephalitis due to type 1 herpes simplex virus (HSV-1). CASE REPORT. A 51-year-old male who had reported the presence of dorsal herpes zoster some days earlier. The patient visited the emergency department after suffering a holocranial oppressive headache and febricula for seven days; 24 hours before admission to hospital, he was suffering from drowsiness and language disorder. The neurological examination revealed stiffness in the back of the neck and dysphasia. An analysis of the cerebrospinal fluid revealed pleocytosis (422 cells/mm(3)) with 98% of mononuclear cells and normal protein and glucose concentration levels in cerebrospinal fluid. Magnetic resonance imaging of the brain and electroencephalogram readings were normal with periodic lateralised epileptiform discharges in the left temporal region. Intravenous acyclovir treatment was initiated, but renal failure meant it had to be changed to oral valaciclovir with clinical resolution and improvement of the liquoral parameters. Polymerase chain reaction in the cerebrospinal fluid was positive for EBV and negative for the other neurotropic viruses. In blood, the serology test for EBV with IgG was positive, while IgM and heterophile antibody tests were negative. CONCLUSIONS. EBV infection can give rise to acute disseminated encephalomyelitis or affect several locations in the central nervous system, especially the cerebellum. Clinical pictures mimicking HSV-1 are less frequent. When encephalitis is related to viral reactivation, precipitating factors can be detected, as in our case. PMID:24203667

Barón, Johanna; Herrero-Velázquez, Sonia; Ruiz-Piñero, Marina; Pedraza, M Isabel; Rojo-Rello, Silvia; Guerrero-Peral, Ángel Luis



Involvement of a neutral glycolipid in differential cell adhesion in the Xenopus blastula.  

PubMed Central

Many different molecular species mediate cell adhesion during embryonic development. These can have either protein or carbohydrate functional groups, which can act in either a homophilic or a heterophilic manner, and often in concert. We report here that a monoclonal antibody, M4B, raised against Xenopus blastomere membranes, inhibits the calcium-dependent adhesion of dissociated blastomeres. M4B maintains its inhibitory effect on adhesion when converted into univalent fragments, and specifically affects calcium-dependent adhesion. The antigen is regulated in both space and time during early development. It is found on cell surfaces throughout the egg to blastula stages, but is more concentrated on cells in the animal and marginal zones of the blastula. It is dramatically downregulated during gastrulation, and becomes largely restricted to gut epithelium by the larval stages. We show also that M4B function is spatially differentiated at the blastula stage, since it inhibits the aggregation of dissociated animal cells to a greater extent than vegetal cells. This membrane antigen may therefore play a role in the differential adhesion observed between different regions of the blastula, and which we presume to underlie the segregation of the primary germ layers during gastrulation. M4B recognizes a complex of plasma membrane glycolipids. Periodate treatment destroys the ability of these glycolipids to react with the antibody, indicating that the epitope resides in the carbohydrate moiety of the glycolipids. Chemical characterization shows that it is a neutral glycolipid, and that the major component is of the glycoglycerolipid, rather than the more common glycosphingolipid class. Blocking experiments with oligosaccharides of defined structure, and antibody crossreactivity show that the M4B antibody does not recognize several known embryonic carbohydrate antigens. These results demonstrate that M4B antibody recognizes a novel group of developmentally regulated glycolipids which function in calcium-dependent cell--cell adhesion in the Xenopus blastula. Images

Turner, A P; Brown, D; Heasman, J; Cook, G M; Evans, J; Vickers, L; Wylie, C C



Infections with Avian Pathogenic and Fecal Escherichia coli Strains Display Similar Lung Histopathology and Macrophage Apoptosis  

PubMed Central

The purpose of this study was to compare histopathological changes in the lungs of chickens infected with avian pathogenic (APEC) and avian fecal (Afecal) Escherichia coli strains, and to analyze how the interaction of the bacteria with avian macrophages relates to the outcome of the infection. Chickens were infected intratracheally with three APEC strains, MT78, IMT5155, and UEL17, and one non-pathogenic Afecal strain, IMT5104. The pathogenicity of the strains was assessed by isolating bacteria from lungs, kidneys, and spleens at 24 h post-infection (p.i.). Lungs were examined for histopathological changes at 12, 18, and 24 h p.i. Serial lung sections were stained with hematoxylin and eosin (HE), terminal deoxynucleotidyl dUTP nick end labeling (TUNEL) for detection of apoptotic cells, and an anti-O2 antibody for detection of MT78 and IMT5155. UEL17 and IMT5104 did not cause systemic infections and the extents of lung colonization were two orders of magnitude lower than for the septicemic strains MT78 and IMT5155, yet all four strains caused the same extent of inflammation in the lungs. The inflammation was localized; there were some congested areas next to unaffected areas. Only the inflamed regions became labeled with anti-O2 antibody. TUNEL labeling revealed the presence of apoptotic cells at 12 h p.i in the inflamed regions only, and before any necrotic foci could be seen. The TUNEL-positive cells were very likely dying heterophils, as evidenced by the purulent inflammation. Some of the dying cells observed in avian lungs in situ may also be macrophages, since all four avian E. coli induced caspase 3/7 activation in monolayers of HD11 avian macrophages. In summary, both pathogenic and non-pathogenic fecal strains of avian E. coli produce focal infections in the avian lung, and these are accompanied by inflammation and cell death in the infected areas.

Horn, Fabiana; Correa, Andre Mendes Ribeiro; Barbieri, Nicolle Lima; Glodde, Susanne; Weyrauch, Karl Dietrich; Kaspers, Bernd; Driemeier, David; Ewers, Christa; Wieler, Lothar H.



Insights on the host stress, fear and growth responses to the deoxynivalenol feed contaminant in broiler chickens.  


Mycotoxins pose an important danger to human and animal health. Poultry feeds are frequently contaminated with deoxynivalenol (DON) mycotoxin. It is thus of great importance to evaluate the effects of DON on the welfare related parameters in poultry industry. In the present study, the effects of contamination of broiler diet with 10 mg DON/kg feed on plasma corticosterone and heterophil to lymphocyte (H/L) ratio as indicators of stress, tonic immobility duration as an index for fear response and growth performance of broiler chickens were studied. In addition, the effect of a microbial feed additive either alone or in combination with DON contamination on these different aspects was also evaluated. The results showed that DON feeding significantly affected the welfare related parameters of broiler chickens. The feeding of DON contaminated diet resulted in an elevation of plasma corticosterone, higher H/L ratio and increased the fear levels as indicated by longer duration of tonic immobility reaction. Furthermore, DON reduced the body weight and body weight gain during the starter phase definitely at the second and third week. However, during grower phase, feeding of DON decreased the body weight at the fourth week and reduced the body gain at the fifth week. Addition of the microbial feed additive, a commercial antidote for DON mycotoxin, was able to overcome DON effects on stress index (H/L ratio), fearfulness and growth parameters of broilers. In conclusion, we showed for the first time that the DON feeding increased the underlying fearfulness and physiological stress responses of broilers and resulted in a reduction in the welfare status as indicated by higher plasma corticosterone, higher H/L ratio and higher fearfulness. Additionally, feeding the microbial feed additive was effective in reducing the adverse effects of DON on the bird's welfare and can improve the performance of broiler chickens. PMID:24498179

Ghareeb, Khaled; Awad, Wageha A; Sid-Ahmed, Omer E; Böhm, Josef



The immunoglobulin domain of the sodium channel ?3 subunit contains a surface-localized disulfide bond that is required for homophilic binding  

PubMed Central

The ? subunits of voltage-gated sodium (Nav) channels possess an extracellular immunoglobulin (Ig) domain that is related to the L1 family of cell-adhesion molecules (CAMs). Here we show that in HEK293 cells, secretion of the free Ig domain of the ?3 subunit is reduced significantly when it is coexpressed with the full-length ?3 and ?1 subunits but not with the ?2 subunit. Using immunoprecipitation, we show that the ?3 subunit can mediate trans homophilic-binding via its Ig domain and that the ?3-Ig domain can associate heterophilically with the ?1 subunit. Evolutionary tracing analysis and structural modeling identified a cluster of surface-localized amino acids fully conserved between the Ig domains of all known ?3 and ?1 sequences. A notable feature of this conserved surface cluster is the presence of two adjacent cysteine residues that previously we have suggested may form a disulfide bond. We now confirm the presence of the disulfide bond in ?3 using mass spectrometry, and we show that its integrity is essential for the association of the full-length, membrane-anchored ?3 subunit with itself. However, selective reduction of this surface disulfide bond did not inhibit homophilic binding of the purified ?3-Ig domain in free solution. Hence, the disulfide bond itself is unlikely to be part of the homophilic binding site. Rather, we suggest that its integrity ensures the Ig domain of the membrane-tethered ?3 subunit adopts the correct orientation for productive association to occur in vivo.—Yereddi, N. R., Cusdin, F. S., Namadurai, S., Packman, L. C., Monie, T. P., Slavny, P., Clare, J. C., Powell, A. J., Jackson, A. P. The immunoglobulin domain of the sodium channel ?3 subunit contains a surface-localized disulfide bond that is required for homophilic binding.

Yereddi, Nikitha R.; Cusdin, Fiona S.; Namadurai, Sivakumar; Packman, Len C.; Monie, Tom P.; Slavny, Peter; Clare, Jeffrey J.; Powell, Andrew J.; Jackson, Antony P.



Hematologic and plasma biochemistry reference intervals of healthy adult barn owls (Tyto alba).  


Hematologic and plasma biochemistry parameters of barn owls (Tyto alba) were studied in collaboration by the Exotic Division of the Faculty of Veterinary Science of the Szent Istvan University and the Eötvös Loránd University, both in Budapest, Hungary. Blood samples were taken from a total of 42 adult barn owls kept in zoos and bird repatriation stations. The following quantitative and qualitative hematologic values were determined: packed cell volume, 46.2 +/- 4%; hemoglobin concentration, 107 +/- 15 g/L; red blood cell count, 3.2 +/- 0.4 x 10(12)/L; white blood cell count, 13.7 +/- 2.7 x 10(9)/L; heterophils, 56.5 +/- 11.5% (7.8 +/- 2 x 10(9)/L); lymphocytes, 40.3 +/- 10.9% (5.5 +/- 1.9 x 10(9)/L); monocytes, 1.8 +/- 2.1% (0.3 +/- 0.3 x 10(9)/ L); eosinophils, 1 +/- 1% (0.1 +/- 0.1 x 10(9)/L); and basophils, 0.6 +/- 0.5% (0.1 +/- 0.1 x 10(9)/L). The following plasma biochemistry values also were determined: aspartate aminotransferase, 272 +/- 43 U/L; L-gamma-glutamyltransferase, 9.5 +/- 4.7 U/L; lipase, 31.7 +/- 11.1 U/L; creatine kinase, 2228 +/- 578 U/L; lactate dehydrogenase, 1702 +/- 475 U/L; alkaline phosphatase, 358 +/- 197 U/L; amylase, 563 +/- 114 U/L; glutamate dehydrogenase, 7.5 +/- 2.5 U/L; total protein, 30.6 +/- 5.3 g/L; uric acid, 428 +/- 102 micromol/L; and bile acids, 43 +/- 18 micromol/L. These results provide reliable reference values for the clinical interpretation of hematologic and plasma biochemistry results for the species. PMID:25055626

Szabo, Zoltan; Klein, Akos; Jakab, Csaba



The effect of feeding rations with different ratios of concentrate to alfalfa hay on blood hematological and biochemical parameters of farmed ostriches (Struthio camelus).  


A two-month trial was conducted to investigate the effects of pelleting rations with 70:30, 65:35, and 60:40 concentrate to high-quality roughage (alfalfa hay) ratios on hematological and biochemical parameters of ostriches. A total of 18 seven-month-old male ostrich chicks, initially weighing an average of 60-70 kg, were distributed into three different outdoor paddocks at a stocking density of six birds per paddock. In the morning, blood collection was made from the wing vein after about 12 h of fasting. All rations resulted in no significant change in hematological parameters. However, with increasing alfalfa inclusion rate, the heterophil/lymphocyte ratio tended to be lower (P = 0.089); whereas the values of hemoglobin, packed cell volume (PCV), and mean corpuscular hemoglobin (MCH) tended to be higher (P values 0.069, 0.072 and 0.094, respectively). In terms of plasma biochemical parameters, increasing the ratio of concentrate to alfalfa hay in ostrich diets resulted in significant (P < 0.05) depression in the values of glucose (up to 20.2 %), total cholesterol (up to 19.2 %), creatine kinase (up to 27.9 %), and aspartate aminotransferase (up to 29.9 %). Additionally, the 65:35 or 60:40 rations caused a decrease in low-density lipoprotein cholesterol level (27.9 and 42 %, respectively; P = 0.008) compared to the 70:40 ration. Without exception, no rations affected the values of leukocyte subsets, total protein, albumin, globulin, uric acid, alanine aminotransferase, gamma glutamyl transferase, triglycerides, high-density lipoprotein cholesterol, and very low density lipoprotein. Based on our results, the concentrate to alfalfa ratio of 60:40 could be suggested as optimum ratio for good health conditions of juvenile ostriches. PMID:23575919

Ghasemi, Hossein Ali; Kazemi-Bonchenari, Mehdi; Khaltabadi-Farahani, Amir Hossein; Khodaei Motlagh, Mahdi



The chick chorioallantoic membrane as an in vivo xenograft model for Burkitt lymphoma  

PubMed Central

Background Burkitt lymphoma (BL) is an aggressive malignancy that arises from B-cells and belongs to the group of Non-Hodgkin lymphomas (NHL). Due to the lack of appropriate in vivo models NHL research is mainly performed in vitro. Here, we studied the use of the chick chorioallantoic membrane (CAM) for the generation of human BL xenograft tumors, which we compared with known characteristics of the human disease. Methods In order to generate experimental BL tumors, we inoculated human BL2B95 and BL2-GFP cells on the CAM. BL2B95 xenograft-tumors were grown for seven days and subsequently analyzed with transmission electron and immunofluorescence microscopy, as well as histological staining approaches. BL2-GFP cells were studied at regular intervals up to seven days, and their metastatic behavior was visualized with intravital immunofluorescence techniques. Results Xenografted BL2B95 cells formed solid tumors in the CAM model with a Ki67-index greater than 90%, preservation of typical tumor markers (CD10, CD19, CD20), a ‘starry sky’ morphology, production of agyrophilic fibers in the stroma, formation of blood and lymphatic vessels and lymphogenic dissemination of BL2B95 to distant sites. We identified macrophages, lymphocytes and heterophilic granulocytes (chick homolog of neutrophils) as the most abundant immune cells in the experimental tumors. BL2-GFP cells could be traced in real-time during their distribution in the CAM, and the first signs for their dissemination were visible after 2-3 days. Conclusions We show that xenografted BL2B95 cells generate tumors in the CAM with a high degree of cellular, molecular and proliferative concord with the human disease, supporting the application of the CAM model for NHL research with a focus on tumor-stroma interactions. Additionally we report that BL2-GFP cells, grafted on the CAM of ex ovo cultured chick embryos, provide a powerful tool to study lymphogenic dissemination in real-time.



Pathology and Pathophysiology of Inhalational Anthrax in a Guinea Pig Model  

PubMed Central

Nonhuman primates (NHPs) and rabbits are the animal models most commonly used to evaluate the efficacy of medical countermeasures against anthrax in support of licensure under the FDA's “Animal Rule.” However, a need for an alternative animal model may arise in certain cases. The development of such an alternative model requires a thorough understanding of the course and manifestation of experimental anthrax disease induced under controlled conditions in the proposed animal species. The guinea pig, which has been used extensively for anthrax pathogenesis studies and anthrax vaccine potency testing, is a good candidate for such an alternative model. This study was aimed at determining the median lethal dose (LD50) of the Bacillus anthracis Ames strain in guinea pigs and investigating the natural history, pathophysiology, and pathology of inhalational anthrax in this animal model following nose-only aerosol exposure. The inhaled LD50 of aerosolized Ames strain spores in guinea pigs was determined to be 5.0 × 104 spores. Aerosol challenge of guinea pigs resulted in inhalational anthrax with death occurring between 46 and 71 h postchallenge. The first clinical signs appeared as early as 36 h postchallenge. Cardiovascular function declined starting at 20 h postexposure. Hematogenous dissemination of bacteria was observed microscopically in multiple organs and tissues as early as 24 h postchallenge. Other histopathologic findings typical of disseminated anthrax included suppurative (heterophilic) inflammation, edema, fibrin, necrosis, and/or hemorrhage in the spleen, lungs, and regional lymph nodes and lymphocyte depletion and/or lymphocytolysis in the spleen and lymph nodes. This study demonstrated that the course of inhalational anthrax disease and the resulting pathology in guinea pigs are similar to those seen in rabbits and NHPs, as well as in humans.

Savransky, Vladimir; Sanford, Daniel C.; Syar, Emily; Austin, Jamie L.; Tordoff, Kevin P.; Anderson, Michael S.; Stark, Gregory V.; Barnewall, Roy E.; Briscoe, Crystal M.; Lemiale-Bierinx, Laurence; Park, Sukjoon; Ionin, Boris



Aerosolized Bacillus anthracis Infection in New Zealand White Rabbits: Natural History and Intravenous Levofloxacin Treatment  

PubMed Central

The natural history for inhalational Bacillus anthracis (Ames strain) exposure in New Zealand white rabbits was investigated to better identify potential, early biomarkers of anthrax. Twelve SPF Bordetella-free rabbits were exposed to 150 LD50 aerosolized B. anthracis spores, and clinical signs, body temperature, complete blood count, bacteremia, and presence of protective antigen in the blood (that is, antigenemia) were examined. The development of antigenemia and bacteremia coincided and preceded both pyrexia and inversion of the heterophil:lymphocyte ratio, an indicator of infection. Antigenemia was determined within 1 h by electrochemiluminescence immunoassay, compared with the 24-h traditional culture needed for bacteremia determination. Rabbits appeared clinically normal until shortly before succumbing to anthrax approximately 47 h after challenge or approximately 22 h after antigenemia, which suggests a relatively narrow therapeutic window of opportunity. To evaluate the therapeutic rabbit model, B. anthracis-exposed rabbits were treated (after determination of antigenemia and later confirmed to be bacteremic) intravenously with the fluoroquinolone antibiotic levofloxacin for 5 d at a total daily dose of 25 or 12.5 mg/kg, resulting in nearly 90% and 70% survival, respectively, to the study end (28 d after challenge). The peak level for 12.5 mg/kg was equivalent to that observed for a 500-mg daily levofloxacin dose in humans. These results suggest that intravenous levofloxacin is an effective therapeutic against inhalational anthrax. Taken together, our findings indicate that antigenemia is a viable and early biomarker for B. anthracis infection that can be used as a treatment trigger to allow for timely intervention against this highly pathogenic disease.

Yee, Steven B; Hatkin, Joshua M; Dyer, David N; Orr, Steven A; Pitt, M Louise M



Epstein-Barr virus and cytomegalovirus in autoimmune diseases: are they truly notorious? A preliminary report.  


To date, it is believed that the origin of autoimmune diseases is one of a multifactorial background. A genetic predisposition, an immune system malfunction or even backfire, hormonal regulation, and environmental factors all play important roles in the pathogenesis of autoimmune diseases. Among these environmental factors, the role of infection is known to be a major one. Epstein-Barr virus (EBV) and cytomegalovirus (CMV) are considered to be notorious as they are consistently associated with multiple autoimmune diseases. A cohort of 1595 serum samples, of 23 different autoimmune disease groups, was screened for evidence of prior infection with EBV and CMV. All samples were screened for antibodies against EBV nuclear antigen-1 (IgG), EBV viral capsid antigen (IgG and IgM), EBV early antigen (IgG), EBV heterophile antibody, and CMV (IgG and IgM) antibodies using Bio-Rad's BioPlex 2200. A new association is proposed between EBV and polymyositis, as results show a significant increase in titers of various EBV target analytes when compared with healthy controls. Our results also support prior information suggesting the association between EBV and multiple autoimmune diseases, including SLE, antiphospholipid syndrome, rheumatoid arthritis, multiple sclerosis, pemphigus vulgaris, giant cell arthritis, Wegener's granulomatosis, and polyarteritis nodosa (PAN). Elevated CMV IgG titers were observed in sera of SLE patients. Our data support the theory that EBV is notoriously associated with many autoimmune diseases. CMV appears to be associated to autoimmune diseases as well, yet establishing this theory requires further investigation. PMID:17894021

Barzilai, O; Sherer, Y; Ram, M; Izhaky, D; Anaya, J M; Shoenfeld, Y



Infectious Mononucleosis at the United States Military Academy. A Prospective Study of a Single Class Over Four Years 1  

PubMed Central

A prospective study of EB virus infections was initiated in July, 1969 in the entering class of 1401 cadets, at the U.S. Military Academy at West Point, N.Y. and continued over 4 yr. On entry 63.5% possessed EBV antibody and 36.5 lacked EBV antibody. The rate of antibody prevalence varied with the geographic area from which the cadet originated. Except in two cadets already ill on first bleeding no evidence of clinical infectious mononucleosis (I.M.) occurred over the 4 yr period in the 890 cadets entering the Academy with EBV antibody. Among 437 cadets without antibody on entry, 54 or 12.4% were infected (seroconverted) in the freshman year; 15 of these had clinical I.M., 12 had suggestive I.M., and 39 had no known mono-like illness. The annual infection rates in susceptible cadets in the second, third, and fourth years were 24.4, 15.1, and 30.8 per 100, respectively. Of 201 cadets infected with EBV over 4 yr only 26.4% were manifested by heterophile positive clinical infectious mononucleosis. Overall, 46% of the 437 cadets entering without EBV antibody became infected over 40 mo of serologic observation; definite clinical infectious mononucleosis developed in 53 cadets, a clinical attack rate of 12.1 per 100 for 4 yr. The EBV infection rate among exposed and susceptible roommates of known cases was no higher than in roommates not so exposed. Elevations of EBV-specific and total IgM occurred during acute illness and disappeared in late convalescence. Total IgG and IgA levels were less commonly elevated. EBV-specific-IgM antibody was demonstrable during the acute illness but was absent 12 mo later. Analysis of EBV infection rates revealed no difference among persons of different ABO blood groups.

Hallee, T. James; Evans, Alfred S.; Niederman, James C.; Brooks, Charles M.; Voegtly, John H.



Functional characterization of chondroitin sulfate proteoglycans of brain: interactions with neurons and neural cell adhesion molecules  

PubMed Central

Ng-CAM and N-CAM are cell adhesion molecules (CAMs), and each CAM can bind homophilically as demonstrated by the ability of CAM-coated beads (Covaspheres) to self-aggregate. We have found that the extent of aggregation of Covaspheres coated with either Ng-CAM or N-CAM was strongly inhibited by the intact 1D1 and 3F8 chondroitin sulfate proteoglycans of rat brain, and by the core glycoproteins resulting from chondroitinase treatment of the proteoglycans. Much higher concentrations of rat chondrosarcoma chondroitin sulfate proteoglycan (aggrecan) core proteins had no significant effect in these assays. The 1D1 and 3F8 proteoglycans also inhibited binding of neurons to Ng-CAM when mixtures of these proteins were adsorbed to polystyrene dishes. Direct binding of neurons to the proteoglycan core glycoproteins from brain but not from chondrosarcoma was demonstrated using an assay in which cell-substrate contact was initiated by centrifugation, and neuronal binding to the 1D1 proteoglycans was specifically inhibited by the 1D1 monoclonal antibody. Different forms of the 1D1 proteoglycan have been identified in developing and adult brain. The early postnatal form (neurocan) was found to bind neurons more effectively than the adult proteoglycan, which represents the C-terminal half of the larger neurocan core protein. Our results therefore indicate that certain brain proteoglycans can bind to neurons, and that Ng-CAM and N-CAM may be heterophilic ligands for neurocan and the 3F8 proteoglycan. The ability of these brain proteoglycans to inhibit adhesion of cells to CAMs may be one mechanism to modulate cell adhesion and migration in the nervous system.



Mycoplasma corogypsi-associated polyarthritis and tenosynovitis in black vultures (Coragyps atratus).  


Three wild American black vultures (Coragyps atratus) were presented to rehabilitation centers with swelling of multiple joints, including elbows, stifles, hocks, and carpal joints, and of the gastrocnemius tendons. Cytological examination of the joint fluid exudate indicated heterophilic arthritis. Radiographic examination in 2 vultures demonstrated periarticular soft tissue swelling in both birds and irregular articular surfaces with subchondral bone erosion in both elbows in 1 bird. Prolonged antibiotic therapy administered in 2 birds did not improve the clinical signs. Necropsy and histological examination demonstrated a chronic lymphoplasmacytic arthritis involving multiple joints and gastrocnemius tenosynovitis. Articular lesions varied in severity and ranged from moderate synovitis and cartilage erosion and fibrillation to severe synovitis, diffuse cartilage ulceration, subchondral bone loss and/or sclerosis, pannus, synovial cysts, and epiphyseal osteomyelitis. No walled bacteria were observed or isolated from the joints. However, mycoplasmas polymerase chain reactions were positive in at least 1 affected joint from each bird. Mycoplasmas were isolated from joints of 1 vulture that did not receive antibiotic therapy. Sequencing of 16S rRNA gene amplicons from joint samples and the mycoplasma isolate identified Mycoplasma corogypsi in 2 vultures and was suggestive in the third vulture. Mycoplasma corogypsi identification was confirmed by sequencing the 16S-23S intergenic spacer region of mycoplasma isolates. This report provides further evidence that M. corogypsi is a likely cause of arthritis and tenosynovitis in American black vultures. Cases of arthritis and tenosynovitis in New World vultures should be investigated for presence of Mycoplasma spp, especially M. corogypsi. PMID:22903399

Van Wettere, A J; Ley, D H; Scott, D E; Buckanoff, H D; Degernes, L A



[Specific immunity and polymorphism of breeding plumage in pied flycatcher (Ficedula hypoleuca) males (Aves: passeriformes)].  


The relationship between the type of melanin-based plumage coloration and the strength of experimentally induced immune response was studied using as an example a pied flycatcher population from the Moscow Region. The plumage of pied flycatcher males exhibits the full spectrum of transitions from contrasting black-and-white to cryptic brownish, the latter being very similar to the coloration of females. In spite of numerous studies, the nature of this polymorphism still remains vague. Unlike many other avian species with monocyclic breeding, a considerable fraction of pied flycatchers combines two energy-consuming productive processes, breeding and molt. During the main experimental treatment we activated the humoral immunity of free-living males in chick-rearing period by injection of nonpathogenic multigenic antigen (sheep red blood cells, SRBC) and estimated the strength of the immune responses after repeated captures in 6-8 days. In addition, after each capture we estimated the numbers of leucocytes (WBC), heterophil to lymphocyte ratios (H/L) and measured night time basal metabolic rates (BMR). Non-molting males of different color types showed the same immune responses. Among molting birds, the strength of the immune response was significantly higher in pale males (morphs 4-7 by Drost's scale) than in bright males with rich melanin-based coloration (morphs 2-3). This difference resulted from two opposite processes. During molting, pale males heightened the antibody titer after immunization, while bright males tended to reduce the strength of immune response. Possibly such an asymmetry in immunocompetence at the first stage of molt reflects the different life strategies of pied flycatcher males - conspicuous birds less commonly combine breeding with molt than cryptic ones. PMID:23136790

Kerimov, A B; Rogovin, K A; Ivankina, E V; Bushuev, A V; Sokolova, O V; Il'ina, T A



Assessment of growth performance, immune responses, serum metabolites, and prevalence of leg weakness in broiler chicks submitted to early-age water restriction.  


The objective of this study was to evaluate the effects of two methods of early-age water restriction on performance, some immune responses, serum metabolites, and prevalence of leg weakness in broiler chicks. One-hundred-eighty-seven-day-old chicks (Ross 308) were allocated to three treatments with four replicates based on a completely randomized design. Treatments included the control group, where birds had free access to water, and water-restricted groups, where access to water was limited to three 2-h periods (WRT1) or a 12-h period (WRT2) per day from days 7 to 17. Chicks in the control group were significantly heavier (P?heterophil to lymphocyte ratio compared to the control group (P?

Toghyani, Mehdi; Toghyani, Majid; Shahryar, Habib Aghdam; Zamanizad, Mehdi



Periodic usage of low-protein methionine-fortified diets in broiler chickens under high ambient temperature conditions: effects on performance, slaughter traits, leukocyte profiles and antibody response  

NASA Astrophysics Data System (ADS)

This study was performed to evaluate the effects of adding methionine supplements to low-protein diets and subsequent re-feeding with a normal diet on the productive performance, slaughter parameters, leukocyte profiles and antibody response in broiler chickens reared under heat stress conditions. During the whole experimental period (6-49 days), the birds were raised in battery cages located in high ambient temperature in an open-sided housing system. A total of 360 6-day-old male chickens were divided into six treatments in six replicates with ten chicks each. Six isoenergetic diets, with similar total sulfur amino acids levels, were formulated to provide 100 and 100 (control), 85 and 100 (85S), 70 and 100 (70S), 85 and 85 (85SG), 70 and 85 (70S85G), and 70 and 70 % (70SG) of National Research Council recommended levels for crude protein during the starter (6-21 day) and grower (22-42 day) periods, respectively. Subsequently, all groups received a diet containing the same nutrients during the finisher period (43-49 day). The results showed that, under heat stress conditions, average daily gain and feed conversion ratio and performance index from day 6 to 49, breast and thigh yields and antibody titer against Newcastle disease in the birds fed diets 85S, 70S and 85SG were similar to those of birds fed control diet, whereas feeding diets 70S85G and 70SG significantly decreased the values of above-mentioned parameters. Additionally, diets 85S, 70S and 85SG significantly decreased mortality rate and heterophil:lymphocyte ratio compared with the control diet. In conclusion, the results indicate that supplementation of methionine to diets 85S, 70S and 85SG, and then re-feeding with a conventional diet is an effective tool to maintain productive performance and to improve health indices and heat resistance in broilers under high ambient temperature conditions.

Ghasemi, Hossein Ali; Ghasemi, Rohollah; Torki, Mehran



Postsynaptic Receptors for Amyloid-? Oligomers as Mediators of Neuronal Damage in Alzheimer's Disease  

PubMed Central

The neurotoxic effect of amyloid-? peptide (A?) over the central synapses has been described and is reflected in the decrease of some postsynaptic excitatory proteins, the alteration in the number and morphology of the dendritic spines, and a decrease in long-term potentiation. Many studies has been carried out to identify the putative A? receptors in neurons, and is still no clear why the A? oligomers only affect the excitatory synapses. A? oligomers bind to neurite and preferentially to the postsynaptic region, where the postsynaptic protein-95 (PSD-95) is present in the glutamatergic synapse, and interacts directly with the N-methyl-D-aspartate receptor (NMDAR) and neuroligin (NL). NL is a postsynaptic protein which binds to the presynaptic protein, neurexin to form a heterophilic adhesion complex, the disruption of this interaction affects the integrity of the synaptic contact. Structurally, NL has an extracellular domain homolog to acetylcholinesterase, the first synaptic protein that was found to interact with A?. In the present review we will document the interaction between A? and the extracellular domain of NL-1 at the excitatory synapse, as well as the interaction with other postsynaptic components, including the glutamatergic receptors (NMDA and mGluR5), the prion protein, the neurotrophin receptor, and the ?7-nicotinic acetylcholine receptor. We conclude that several A? oligomers receptors exist at the excitatory synapse, which could be the responsible for the neurotoxic effect described for the A? oligomers. The characterization of the interaction between A? receptors and A? oligomers could help to understand the source of the neurologic damage observed in the brain of the Alzheimer’s disease patients.

Dinamarca, Margarita C.; Rios, Juvenal A.; Inestrosa, Nibaldo C.



Habitat and sex differences in physiological condition of breeding Southwestern Willow Flycatchers (Empidonax traillii extimus)  

USGS Publications Warehouse

The Southwestern Willow Flycatcher (Empidonax traillii extimus; hereafter "flycatcher") is a federally listed endangered species that breeds in densely vegetated riparian habitats dominated by native and exotic plants, including introduced monotypic saltcedar (Tamarix ramosissima). Some workers have theorized that saltcedar is unsuitable habitat for the flycatcher, primarily because it generally supports a smaller and less diverse invertebrate community (the flycatcher's food base) than native habitats (e.g. Salix spp.). However, differences in insect communities between native and saltcedar habitats are not proof that saltcedar habitats are inferior. The only way to evaluate whether the habitats differ in dietary or energetic quality is to document actual food limitation or its manifestations. Measurements of an individual's body condition and metabolic state can serve as indicators of environmental stressors, such as food limitation and environmental extremes. We captured 130 flycatchers breeding in native and saltcedar habitats in Arizona and New Mexico and measured 12 variables of physiological condition. These variables included body mass, fat level, body condition index, hematocrit, plasma triglycerides, plasma free fatty acids and glycerol, plasma glucose and beta-hydroxybutyrate, plasma uric acid, total leukocyte count, and heterophil-to-lymphocyte ratio. We found substantial sex-based differences in the condition of male and female flycatchers. Ten of the 12 measures of physiological condition differed significantly between the sexes. In all cases where male and female condition differed (except mass), the differences suggest that males were in poorer condition than females. We found few habitat-based differences in flycatcher condition. Only 3 of the 12 physiological condition indices differed significantly between habitats. Our data show that, at least in some parts of the flycatcher's range, there is no evidence that flycatchers breeding in saltcedar habitats exhibit poorer nutritional condition or are suffering negative physiological affects. ?? The American Ornithologists' Union, 2005.

Owen, J. C.; Sogge, M. K.; Kern, M. D.



Types of adrenocorticoids and their effect on organophosphorus-induced delayed neuropathy in chickens.  


The present study examined the effects of a glucocorticoid and a mineralocorticoid on organophosphorus-induced delayed neuropathy (OPIDN) as previous investigations have indicated that an endogenous steroid with both properties could alter this syndrome in chickens. The glucocorticoid triamcinolone and the mineralocorticoid deoxycorticosterone were provided in the diet beginning 1 day before and continuing 10 days after triortho-tolyl phosphate (TOTP, 360 mg/kg po), phenyl saligenin phosphate (PSP, 2.5 mg/kg im), and diisopropyl phosphorofluoridate (DFP, 1 mg/kg sc). In a manner similar to that seen with corticosterone, a low concentration (0.1 ppm) of triamcinolone reduced and a high concentration (10 ppm) exacerbated clinical signs. Concentrations of deoxycorticosterone under 80 ppm also partially delayed or ameliorated ataxia induced by TOTP, PSP, and DFP, but a combination of 0.1 ppm triamcinolone and 80 ppm deoxycorticosterone was not more effective than triamcinolone alone. Peripheral nerve damage was noted in all chickens given organophosphorus compounds, whether or not they had been given corticoids. Both steroids induced hydroxylase activity, but effects on most other enzyme systems examined were unremarkable. High concentrations of triamcinolone (10 ppm) could, however, also reduce liver cytochrome P450 levels and liver cholinesterase activity. Exacerbation of OPIDN was most notable in chickens under highest stress, as indicated by elevated heterophil-to-lymphocyte ratios. The clinical, pathological, biochemical, and hematological indices of exposure to adrenocorticoids and agents inducing OPIDN in chickens were, therefore, similar for both a synthetic glucocorticoid and the endogenous steroid corticosterone. PMID:3341034

Ehrich, M; Jortner, B S; Gross, W B



West Nile virus in raptors from Virginia during 2003: clinical, diagnostic, and epidemiologic findings.  


Sixty-one birds of prey admitted to The Wildlife Center of Virginia (WCV; Waynesboro, Virginia, USA) from June to November 2003 were tested for West Nile virus (WNV) infection. Choanal and/or cloacal swabs were obtained and submitted to Virginia's Division of Consolidated Laboratory Services (Richmond, Virginia, USA) for analysis with real-time reverse transcriptase polymerase chain reaction (RT-PCR). Forty birds of prey were positive for WNV by RT-PCR. Five avian families and nine species of raptors were represented, with great horned owls (Bubo virginianus) and red-tailed hawks (Buteo jamaicensis) most frequently affected. Presenting clinical signs were consistent with previous reports of WNV infection in raptors; however, these differed between species. Of WNV positive birds, nonspecific signs of illness were the most common clinical findings, particularly in red-tailed hawks; signs included dehydration (n = 20), emaciation (n = 18), and depression (n = 15). Neurologic abnormalities were frequently identified, especially in great horned owls, and included head tremors (n = 17), ataxia (n = 13), head incoordination (n = 7), torticollis (n = 3), nystagmus (n = 3), and head tilt (n = 3). Great horned owls exhibited anemia and leukocytosis with heterophilia, eosinophilia, and monocytosis consistent with chronic inflammation. Red-tailed hawks were anemic with a heterophilic leukocytosis and regenerative left shift. The majority of WNV cases occurred during August and September; there was a marked increase in the number of raptors admitted to WCV during these months followed by a marked decrease during October, November, and December. This pattern differed from mean monthly admissions during the previous 10 years and suggests a negative impact on local raptor populations. The effects of WNV on avian populations are largely unknown; however, because of their ecological importance, further investigation of the effects of WNV on raptor populations is warranted. PMID:16870856

Joyner, Priscilla H; Kelly, Sean; Shreve, Allison A; Snead, Sarah E; Sleeman, Jonathan M; Pettit, Denise A



[Examination of the patient serum presenting a CEA false high value after cancer immuno-cell therapy].  


False positive elevation of carcinoembryonic antigen (CEA) was observed in a 65-year-old woman who was treated with dendritic cell therapy (DCT) and activated lymphocyte therapy (ALT) for intrahepatic cholangiocarcinoma. Three months after the initiation of these therapies, her CEA value measured by AIA (TOSOH) began to increase without any evidence of worsening of cholangiocarcinoma. CEA was measured by several different methods, and only the result measured by AIA was high, indicating the presence of a false positive phenomenon. To clarify this phenomenon, we evaluated the patient's serum precisely. Gel filtration chromatography of her serum showed that CEA was detected in the elution fraction of IgG, which was different from the reference samples. Furthermore, this peak disappeared after incubation of patient's CEA and HBR-1. The immunoglobulin absorption test revealed that CEA value was decreased only after absorption of IgG and absorption tests using HBR-1 and MAK-absorbents showed a dramatic decrease in CEA value. These findings indicated the presence of IgG type human anti-mouse antibodies (HAMA), which interfered the measurement by AIA. Although we could not identify the reason why HAMA was produced in this patient, the facts that the false positive phenomenon was observed after the initiation of DCT and ALT, and that CEA value decreased after theses therapies were discontinued, indicated that immuno-modulaton by DCT and ALT may have a close relationship to HAMA production. It was probable that DCT and ALT activated preexisting heterophile-antibody-producing cells, which stimulated HAMA production. The incidence of such false positive reaction of CEA by HAMA in patients with DCT and ALT was low, but as the number of the patient with immuno-cell therapy increases, the incidence of such phenomenon surely increases. Because HAMA reacts to all types of immunoassay, careful attention should be paid to the evaluation of laboratory findings in patients undergoing with such immuno-cell therapies. PMID:21942086

Abe, Masaki; Matsuura, Tomokazu; Hyoki, Miyuki; Abe, Ikurou; Tanigawa, Keishi; Kaito, Ken



Effects of different levels of organic and inorganic chromium on growth performance and immunocompetence of broilers under heat stress.  


This experiment was carried out to investigate the effects of different levels of organic and inorganic chromium on the performance, immune function and some serum mineral concentrations of broilers under heat stress condition (23.9-37°C cycling). A total of 150 one-day-old broiler chicks according to a completely randomized design were assigned into five treatment groups. Each treatment consisted of three replicates and each replicate contained ten chicks. Chicks were fed on corn-soybean meal basal diets with added different concentrations of chromium (0, 600 and 1,200 ?g kg(-1) chromium chloride or 600 and 1,200 ?g/kg chromium L-methionine) from 1 to 49 days of age. Humoral immunity was assessed by intravenous injection of 7% sheep red blood cell (SRBC) followed by evaluation of serum for antibody titers in primary and secondary responses. Cell-mediated immunity was assessed by the cutaneous basophil hypersensitivity (CBH) test to phytohemagglutinin (PHA)-P at day 32 and PHA-M at day 48. Heterophil/lymphocyte (H/L) ratio was also measured as a reliable indicator of stress. The body mass, feed intake and conversion ratio were not influenced by dietary chromium (P > 0.05). Dietary supplementation of both organic and inorganic chromium significantly increased primary and secondary antibody responses (P < 0.01), and also improved H/L ratio (P < 0.05), CBH response (P < 0.01) as well as relative weights of thymus (P < 0.05) and spleen (P < 0.01). Both dietary organic and inorganic chromium caused an increase in serum concentrations of Cr and Zn (P < 0.01), but decreased the serum concentration of Cu (P < 0.01). These results suggest that supplemental chromium especially in organic form offers a good management practice to reduce heat stress-related depression in immunocompetence of broiler chicks. PMID:22127829

Ghazi, Sh; Habibian, M; Moeini, M M; Abdolmohammadi, A R



Reference intervals for plasma biochemical and hematologic measures in loggerhead sea turtles (Caretta caretta) from Moreton Bay, Australia.  


Biochemical and hematologic reference intervals have been reported for loggerhead sea turtles (Caretta caretta, Linnaeus 1758), but low sample numbers and simple statistical analyses have constrained their diagnostic usefulness. During June 2007-May 2008, 101 loggerhead sea turtles in Moreton Bay, Queensland, Australia, were captured by hand from boats; clinically assessed to determine health status; blood was sampled; and biochemical and hematologic variables were measured. Of these turtles, 66 were classified as clinically healthy and 23 as unhealthy. Reference intervals were calculated using data from clinically healthy turtles. Of the clinically unhealthy turtles, 82 and 45% had at least one biochemical and hematologic result, respectively, outside of at least one of the calculated intervals. However, only low proportions of unhealthy loggerhead sea turtles had abnormal results for each variable. The highest percentage of unhealthy turtles that were outside at least one estimated reference interval was 35%, for thrombocyte counts. Neither sex nor maturity category (mature versus large immature) influenced the risk of being clinically unhealthy. These are the first plasma biochemical and hematologic reference intervals reported for loggerhead sea turtles from the southwestern Pacific Ocean. We conclude that, for loggerhead sea turtles in Moreton Bay, separate reference intervals are required for mature and immature turtles for thrombocyte counts and for male and female turtles for lymphocyte, heterophil, and total white cell counts; otherwise, a single reference interval can be used regardless of age or sex. When estimating reference intervals in loggerhead sea turtles, it is desirable to use both methods for calculating reference intervals used in this study because intervals can differ substantially between methods for some variables. Joint interpretation using reference intervals from both methods allows the categorization of results as "normal," "suspect," or "abnormal." PMID:20688679

Flint, Mark; Morton, John M; Limpus, Colin J; Patterson-Kane, Janet C; Mills, Paul C



Plasmin-sensitive dibasic sequences in the third fibronectin-like domain of L1-cell adhesion molecule (CAM) facilitate homomultimerization and concomitant integrin recruitment.  


L1 is a multidomain transmembrane neural recognition molecule essential for neurohistogenesis. While moieties in the immunoglobulin-like domains of L1 have been implicated in both heterophilic and homophilic binding, the function of the fibronectin (FN)-like repeats remains largely unresolved. Here, we demonstrate that the third FN-like repeat of L1 (FN3) spontaneously homomultimerizes to form trimeric and higher order complexes. Remarkably, these complexes support direct RGD-independent interactions with several integrins, including alpha(v)beta(3) and alpha(5)beta(1). A pep- tide derived from the putative C-C' loop of FN3 (GSQRKHSKRHIHKDHV(852)) also forms trimeric complexes and supports alpha(v)beta(3) and alpha(5)beta(1) binding. Substitution of the dibasic RK(841) and KR(845) sequences within this peptide or the FN3 domain limited multimerization and abrogated integrin binding. Evidence is presented that the multimerization of, and integrin binding to, the FN3 domain is regulated both by conformational constraints imposed by other domains and by plasmin- mediated cleavage within the sequence RK( downward arrow)HSK( downward arrow)RH(846). The integrin alpha(9)beta(1), which also recognizes the FN3 domain, colocalizes with L1 in a manner restricted to sites of cell-cell contact. We propose that distal receptor ligation events at the cell-cell interface may induce a conformational change within the L1 ectodomain that culminates in receptor multimerization and integrin recruitment via interaction with the FN3 domain. PMID:10871287

Silletti, S; Mei, F; Sheppard, D; Montgomery, A M



Cell autonomy of DSCAM function in retinal development  

PubMed Central

Cell adhesion molecules (CAMs) provide identifying cues by which neural architecture is sculpted. The Down Syndrome Cell Adhesion Molecule (DSCAM) is required for many neurodevelopmental processes in different species and also has several potential mechanisms of activity, including homophilic adhesion, homophilic repulsion and heterophilic interactions. In the mouse retina, Dscam is expressed in many, but not all neuronal subtypes. Mutations in Dscam cause the fasciculation of dendrites of neighboring homotypic neurons, indicating a role in self-avoidance among cells of a given type, a disruption of the non-random patterning of their cell bodies, and a decrease in developmental cell death in affected cell populations. In order to address how DSCAM facilitates retinal pattering, we developed a conditional allele of Dscam to use alongside existing Dscam mutant mouse strains. Conditional deletion of Dscam reproduces cell spacing, cell number and dendrite arborization defects. Inducible deletion of Dscam and retinal ganglion cell depletion in Brn3b mutant retinas both indicate that these DSCAM-mediated phenotypes can occur independently. In chimeric retinas, in which wild type and Dscam mutant cells are comingled, Dscam mutant cells entangle adjacent wild type cells of the same type, as if both cells were lacking Dscam, consistent with DSCAM-dependent cell spacing and neurite arborization being mediated through homophilic binding cell-to-cell. Deletion of Dscam in specific cell types causes cell-type-autonomous cell body spacing defects, indicating that DSCAM mediates arborization and spacing by acting within given cell types. We also examine the cell autonomy of DSCAM in laminar stratification and find that laminar disorganization can be caused in a non-cell autonomous fashion. Finally, we find Dscam dosage-dependent defects in developmental cell death and amacrine cell spacing, relevant to the increased cell death and other disorders observed in Down syndrome mouse models and human patients, in which Dscam is present in three copies.

Fuerst, Peter G.; Bruce, Freyja; Rounds, Ryan P.; Erskine, Lynda; Burgess, Robert W.



Evaluation of a stress model induced by dietary corticosterone supplementation in broiler breeders: effects on egg yolk corticosterone concentration and biochemical blood parameters.  


1. This study aimed to evaluate a stress model induced by corticosterone (CORT) supplementation in the diet of broiler breeder hens. 2. A total of 60 Ross broiler breeder hens at 29 weeks of age were randomly divided into 4 groups with 15 hens each. The first group served as the control. The rest of the hens were given 1, 1.5 or 2 mg of CORT/hen/d (CORT1, CORT1.5 and CORT2, respectively) for 7 d. Concentrations of yolk CORT, plasma uric acid, glucose, cholesterol, creatine kinase, heterophil (H):lymphocyte (L) ratio and duration of tonic immobility (TI) were measured at d 3, 5 and 7 of CORT supplementation. The same measurements were repeated at 3, 5 and 7 d after CORT was withdrawn from the diet. 3. There were no significant CORT dose effect on yolk CORT and plasma glucose concentrations. Higher plasma uric acid and H:L ratio was obtained for CORT1.5 and CORT2 than for CORT1. From 3 to 7 d of dietary CORT supplementation, yolk CORT and plasma uric acid concentrations and H:L ratio increased whereas plasma glucose concentration decreased. After CORT was withdrawn from the diet, the H:L ratio remained elevated. The duration of TI and plasma creatine kinase concentration did not change during and after CORT supplementation. 4. Yolk CORT concentration was correlated with plasma uric acid concentration during CORT supplementation. 5. The results suggest that dietary CORT supplementation could be used as a stress model and to evaluate hormone-mediated maternal effects in broiler breeder hens. PMID:24397505

Babacano?lu, E; Yalçin, S; Uysal, S



Changes in heat shock protein 70, blood parameters, and fear-related behavior in broiler chickens as affected by pleasant and unpleasant human contact.  


An experiment was conducted to determine the effects of combining both pleasant and unpleasant contacts with human beings on physiology and behavior of broiler chickens. Birds were subjected to the following treatments: (i) received no physical or visual contact with humans (control); (ii) from d 1 to 28, chicks were individually stroked gently for 30 s once daily (PL); (iii) from d 1 to 28, chicks were picked up individually, suspended by both legs, exposed to recorded noise, and swung gently for 15 s once daily (UNPL); (iv) from d 1 to 14 and from d 15 to 28, chicks were subjected to PL and UNPL, respectively (PL-UNPL); and (v) from d 1 to 14 and from d 15 to 28, chicks were subjected to UNPL and PL, respectively (UNPL-PL). On d 42, birds from each treatment group were road-transported for 3 h. Heat shock protein (hsp) 70 expression, plasma levels of corticosterone, serum creatine kinase concentration, heterophil/lymphocyte ratios (HLR), and tonic immobility duration were determined pre- and posttransit. There were significant (P < 0.05) duration of transportation × human contact treatment interactions for HLR and hsp 70 density. Following transit, the PL chicks had significantly (P < 0.05) lower HLR and greater hsp 70 density than the other groups. The corticosterone of PL and UNPL chicks were lower than their control, PL-UNPL, and UNPL-PL counterparts. The PL and PL-UNPL treatments were effective in shortening tonic immobility duration significantly (P < 0.05). Except for UNPL-PL, the serum creatine kinase activity of PL was significantly lower than the other groups. In conclusion, subjecting birds to pleasant human contact reduced stress and fear reactions to transportation by enhancing the ability to express hsp 70 in the brain. Unpleasant human contact had adverse effect on the birds' response to transportation. Early age pleasant experience with humans failed to negate the adverse effects of subsequent unpleasant contact. PMID:23243228

Al-Aqil, A; Zulkifli, I; Hair Bejo, M; Sazili, A Q; Rajion, M A; Somchit, M N



In tight junctions, claudins regulate the interactions between occludin, tricellulin and marvelD3, which, inversely, modulate claudin oligomerization.  


Tight junctions seal the paracellular cleft of epithelia and endothelia, form vital barriers between tissue compartments and consist of tight-junction-associated marvel proteins (TAMPs) and claudins. The function of TAMPs and the interaction with claudins are not understood. We therefore investigated the binding between the TAMPs occludin, tricellulin, and marvelD3 and their interaction with claudins in living tight-junction-free human embryonic kidney-293 cells. In contrast to claudins and occludin, tricellulin and marvelD3 showed no enrichment at cell-cell contacts indicating lack of homophilic trans-interaction between two opposing cell membranes. However, occludin, marvelD3 and tricellulin exhibited homophilic cis-interactions, along one plasma membrane, as measured by fluorescence resonance energy transfer. MarvelD3 also cis-interacted with occludin and tricellulin heterophilically. Classic claudins, such as claudin-1 to -5 may show cis-oligomerization with TAMPs, whereas the non-classic claudin-11 did not. Claudin-1 and -5 improved enrichment of occludin and tricellulin at cell-cell contacts. The low mobile claudin-1 reduced the membrane mobility of the highly mobile occludin and tricellulin, as studied by fluorescence recovery after photobleaching. Co-transfection of claudin-1 with TAMPs led to changes of the tight junction strand network of this claudin to a more physiological morphology, depicted by freeze-fracture electron microscopy. The results demonstrate multilateral interactions between the tight junction proteins, in which claudins determine the function of TAMPs and vice versa, and provide deeper insights into the tight junction assembly. PMID:23203797

Cording, Jimmi; Berg, Johanna; Käding, Nadja; Bellmann, Christian; Tscheik, Christian; Westphal, Julie K; Milatz, Susanne; Günzel, Dorothee; Wolburg, Hartwig; Piontek, Jörg; Huber, Otmar; Blasig, Ingolf Ernst



Altered Functionality of Anti-Bacterial Antibodies in Patients with Chronic Hepatitis C Virus Infection  

PubMed Central

Background Using comparative glycoproteomics, we have previously identified a glycoprotein that is altered in both amount and glycosylation as a function of liver cirrhosis. The altered glycoprotein is an agalactosylated (G0) immunoglobulin G molecule (IgG) that recognizes the heterophilic alpha-gal epitope. Since the alpha gal epitope is found on gut enterobacteria, it has been hypothesized that anti-gal antibodies are generated as a result of increased bacterial exposure in patients with liver disease. Methods The N-linked glycosylation of anti-gal IgG molecules from patients with fibrosis and cirrhosis was determined and the effector function of anti-bacterial antibodies from over 100 patients examined. In addition, markers of microbial exposure were determined. Results Surprisingly, the subset of agalactosylated anti-gal antibodies described here, was impaired in their ability to mediate complement mediated lysis and inhibited the complement-mediated destruction of common gut bacteria. In an analysis of serum from more than 100 patients with liver disease, we have shown that those with increased levels of this modified anti-gal antibody had increased levels of markers of bacterial exposure. Conclusions Anti-gal antibodies in patients with liver cirrhosis were reduced in their ability to mediate complement mediated lysis of target cells. As bacterial infection is a major complication in patients with cirrhosis and bacterial products such as LPS are thought to play a major role in the development and progression of liver fibrosis, this finding has many clinical implications in the etiology, prognosis and treatment of liver disease.

Lamontagne, Anne; Long, Ronald E.; Comunale, Mary Ann; Hafner, Julie; Rodemich-Betesh, Lucy; Wang, Mengjun; Marrero, Jorge; Di Bisceglie, Adrian M.; Block, Timothy; Mehta, Anand



Serological diagnosis of Epstein-Barr virus infection by novel ELISAs based on recombinant capsid antigens p23 and p18.  


A new pair of Epstein-Barr virus ELISAs (Biotest Anti-EBV VCA IgG and VCA IgM ELISA) was evaluated for usefulness for routine diagnosis of acute EBV infections. The ELISAs are based on two viral capsid antigens (VCA), p23 (BLRF2, full-length) and p18 (BFRF3, carboxy-half), that are combined by autologous gene fusion. In total, 179 sera were tested in direct comparison with classical VCA immunofluorescence assays (IFA). With the help of clinical data and additional reference serology, i.e., heterophile antibodies, anti-EA IgG (IFA) and anti-EBNA-1 IgG (ELISA), the patients were divided into the following categories: seronegatives (46), acute primary infections (67), previous infections (39), suspected reactivations (20) and constellations with intermediate serological patterns (7). The VCA IgG and VCA IgM ELISAs showed overall agreement to IFA of 95.0% and 94.4%, respectively. The calculated analytical performance (sensitivity; specificity) of VCA IgG and VCA IgM was 94.0%; 97.8% and 97.1%; 96.5%, respectively. A certain delay in seroconversion of anti-p23-p18 IgG may account for a significant difference in sensitivity of the VCA IgG ELISA between primary (88.4%) and previous infections (100%). In summary, the new recombinant VCA ELISAs yielded good correlation to VCA IFA and in combination with EBNA-1 IgG allow rapid, sensitive, and specific diagnosis of infectious mononucleosis or EBV immune status in general. PMID:11241457

Färber, I; Hinderer, W; Rothe, M; Lang, D; Sonneborn, H H; Wutzler, P



Expression of Mel-CAM in implantation site intermediate trophoblastic cell line, IST-1, limits its migration on uterine smooth muscle cells.  


An immortalized implantation site intermediate trophoblastic cell line, IST-1, was established from a human placenta of 7 weeks gestation. IST-1 cells phenotypically resembled the implantation site intermediate trophoblastic cells in situ and expressed Mel-CAM (MUC 18 or CD146). Mel-CAM is a cell adhesion molecule belonging to the immunoglobulin gene superfamily. It is involved in heterophilic cell-cell adhesion and plays a role in several biological processes including tumor progression. We have previously shown that Mel-CAM was highly expressed in the intermediate (extravillous) trophoblast in the human implantation site. In this study we determined the function of Mel-CAM in the interaction of trophoblast and uterine smooth muscle in the implantation site. IST-1 cells failed to adhere to immobilized recombinant Mel-CAM in solid phase whereas the uterine smooth muscle cells did. The presence of the putative Mel-CAM ligand in smooth muscle cells was further supported by the finding that Mel-CAM-transfected but not the mock-transfected U937 leukemia cells bind to the confluent monolayer of uterine smooth muscle cells. IST-1 cells attached efficiently to the monolayer of the uterine smooth muscle cells and acquired a spindle-shaped morphology simulating smooth muscle cells. The cell binding was only marginally affected by Mel-CAM blocking antibodies. However, Mel-CAM blocking antibodies and recombinant Mel-CAM promoted cell migration from IST-1 cell spheroids on the smooth muscle monolayer. Taken together, our results suggest that IST-1 cells express Mel-CAM but not the putative Mel-CAM ligand. In contrast, the uterine smooth muscle cells express the putative Mel-CAM ligand which binds to Mel-CAM on the surface of the IST-1 cells. The interaction between Mel-CAM and its putative ligand confers a stationary phenotype for trophoblastic cells. These observations are consistent with an important role for Mel-CAM in limiting trophoblastic migration within the myometrium in the implantation site. PMID:9701564

Shih, I; Wang, T; Wu, T; Kurman, R J; Gearhart, J D



Mathematical Modeling of Sub-Cellular Asymmetry of Fat-Dachsous Heterodimer for Generation of Planar Cell Polarity  

PubMed Central

Planar Cell Polarity (PCP) is an evolutionarily conserved characteristic of animal tissues marked by coordinated polarization of cells or structures in the plane of a tissue. In insect wing epithelium, for instance, PCP is characterized by en masse orientation of hairs orthogonal to its apical-basal axis and pointing along the proximal-distal axis of the organ. Directional cue for PCP has been proposed to be generated by complex sets of interactions amongst three proteins - Fat (Ft), Dachsous (Ds) and Four-jointed (Fj). Ft and Ds are two atypical cadherins, which are phosphorylated by Fj, a Golgi kinase. Ft and Ds from adjacent cells bind heterophilically via their tandem cadherin repeats, and their binding affinities are regulated by Fj. Further, in the wing epithelium, sub-cellular levels of Ft-Ds heterodimers are seen to be elevated at the distal edges of individual cells, prefiguring their PCP. Mechanisms generating this sub-cellular asymmetry of Ft-Ds heterodimer in proximal and distal edges of cells, however, have not been resolved yet. Using a mathematical modeling approach, here we provide a framework for generation of this sub-cellular asymmetry of Ft-Ds heterodimer. First, we explain how the known interactions within Ft-Ds-Fj system translate into sub-cellular asymmetry of Ft-Ds heterodimer. Second, we show that this asymmetric localization of Ft-Ds heterodimer is lost when tissue-level gradient of Fj is flattened, or when phosphorylation of Ft by Fj is abolished, but not when tissue-level gradient of Ds is flattened or when phosphorylation of Ds is abrogated. Finally, we show that distal enrichment of Ds also amplifies Ft-Ds asymmetry. These observations reveal that gradient of Fj expression, phosphorylation of Ft by Fj and sub-cellular distal accumulation of Ds are three critical elements required for generating sub-cellular asymmetry of Ft-Ds heterodimer. Our model integrates the known experimental data and presents testable predictions for future studies.

Kumar, Amit; Sinha, Pradip



Thermal manipulation of the embryo modifies the physiology and body composition of broiler chickens reared in floor pens without affecting breast meat processing quality.  


Selection in broiler chickens has increased muscle mass without similar development of the cardiovascular and respiratory systems, resulting in limited ability to sustain high ambient temperatures. The aim of this study was to determine the long-lasting effects of heat manipulation of the embryo on the physiology, body temperature (Tb), growth rate and meat processing quality of broiler chickens reared in floor pens. Broiler chicken eggs were incubated in control conditions (37.8°C, 56% relative humidity; RH) or exposed to thermal manipulation (TM; 12 h/d, 39.5°C, 65% RH) from d 7 to 16 of embryogenesis. This study was planned in a pedigree design to identify possible heritable characters for further selection of broiler chickens to improve thermotolerance. Thermal manipulation did not affect hatchability but resulted in lower Tb at hatching and until d 28 post-hatch, with associated changes in plasma thyroid hormone concentrations. At d 34, chickens were exposed to a moderate heat challenge (5 h, 32°C). Greater O2 saturation and reduced CO2 partial pressure were observed (P < 0.05) in the venous blood of TM than in that of control chickens, suggesting long-term respiratory adaptation. At slaughter age, TM chickens were 1.4% lighter and exhibited 8% less relative abdominal fat pad than controls. Breast muscle yield was enhanced by TM, especially in females, but without significant change in breast meat characteristics (pH, color, drip loss). Plasma glucose/insulin balance was affected (P < 0.05) by thermal treatments. The heat challenge increased the heterophil/lymphocyte ratio in controls (P < 0.05) but not in TM birds, possibly reflecting a lower stress status in TM chickens. Interestingly, broiler chickens had moderate heritability estimates for the plasma triiodothyronine/thyroxine concentration ratio at d 28 and comb temperature during the heat challenge on d 34 (h(2) > 0.17). In conclusion, TM of the embryo modified the physiology of broilers in the long term as a possible adaptation for heat tolerance, without affecting breast meat quality. This study highlights the value of 2 new heritable characters involved in thermoregulation for further broiler selection. PMID:23736053

Loyau, T; Berri, C; Bedrani, L; Métayer-Coustard, S; Praud, C; Duclos, M J; Tesseraud, S; Rideau, N; Everaert, N; Yahav, S; Mignon-Grasteau, S; Collin, A



Health of tree swallows (Tachycineta bicolor) nesting in pesticide-sprayed apple orchards in Ontario, Canada. I. Immunological parameters.  


The degree of pesticide exposure and its effects on the immune system and its development were determined in 16-d-old tree swallow (Tachycineta bicolor) chicks from 4 sprayed apple orchards and three nonsprayed sites in southern Ontario, Canada, during 1994-1995. Persistent contaminant residues were measured in tree swallow eggs and in each chick hepatic ethoxyresorufin O-deethylase (EROD) activity; body, immune organ, and liver masses; lymphocyte blastogenesis response; respiratory burst and phagocytic responses; hemarological evaluation; and histological development of thymus, bursa of fabricius, and spleen were determined. Chemicals sprayed on apple orchards were mainly ethylene bisdithiocarbamate and myclobutanil fungicides and organophosphorus, carbamate, and synthetic pyrethroid insecticides. During the period between oviposition of the first egg in each nest to d 16 after hatching, individual nests in orchards were exposed to between 4 and 11 individual chemical applications and up to 3 mixtures of pesticide sprays. Concentrations of pesticides, polychlorinated biphenyls (PCBs), and lead and arsenic residues in tree swallow eggs and liver were low and not variable among sites except p,p'-DDE, which was as high as 2.29 microg/g wet weight in eggs. EROD activity was not different among sites. Organochlorine and trace metal residues and EROD activity were not correlated with any immune parameter. In sprayed birds, we found a significantly increased blastogenic response to pokeweed mitogen (12.5 microg/ml). However, nests were initiated over a period of several weeks and we also found changes in other tree swallow immune parameters that were related to the date of chick collection. Hematological parameters, bursal and thymic masses, phagocytic response, and thymic development were all correlated with the day the chicks were 16 d of age. After accounting for the collection date of birds from each nest, we found cell proliferation in the cortex and delayed thymic involution correlated positively with increasing spray exposure. We also found that birds in sprayed orchards were slightly anemic compared to birds from nonsprayed sites, and there were smaller bursal masses and an increase in relative heterophil concentrations in the sprayed orchard birds. The local inflammation may have been caused by trematode parasite infections, although pesticide exposure also correlated positively with these parameters. This is the first study of the immunology and effects of current pesticide exposures in wild passerines; therefore it is difficult to predict the long-term consequences of the apparent stimulated immune systems in sprayed birds. However, some environmental contaminants that overtly stimulate the immune system in mammals have induced hypersensitivity and/or autoimmunity. Therefore we speculate that these effects are possible in tree swallows. PMID:9885997

Bishop, C A; Boermans, H J; Ng, P; Campbell, G D; Struger, J



Differential contributions of Ng-CAM and N-CAM to cell adhesion in different neural regions  

PubMed Central

Individual neurons can express both the neural cell adhesion molecule (N-CAM) and the neuron-glia cell adhesion molecule (Ng-CAM) at their cell surfaces. To determine how the functions of the two molecules may be differentially controlled, we have used specific antibodies to each cell adhesion molecule (CAM) to perturb its function, first in brain membrane vesicle aggregation and then in tissue culture assays testing the fasciculation of neurite outgrowths from cultured dorsal root ganglia, the migration of granule cells in cerebellar explants, and the formation of histological layers in the developing retina. Our strategy was initially to delineate further the binding mechanisms for each CAM. Antibodies to Ng-CAM and N-CAM each inhibited brain membrane vesicle aggregation but the binding mechanisms of the two CAMs differed. As expected from the known homophilic binding mechanism of N-CAM, anti-N- CAM-coated vesicles did not co-aggregate with uncoated vesicles. Anti- Ng-CAM-coated vesicles readily co-aggregated with uncoated vesicles in accord with a postulated heterophilic binding mechanism. It was also shown that N-CAM was not a ligand for Ng-CAM. In contrast to assays with brain membrane vesicles, cellular systems can reveal functional differences for each CAM reflecting its relative amount (prevalence modulation) and location (polarity modulation). Consistent with this, each of the three cellular processes examined in vitro was preferentially inhibited only by anti-N-CAM or by anti-Ng-CAM antibodies. Both neurite fasciculation and the migration of cerebellar granule cells were preferentially inhibited by anti-Ng-CAM antibodies. Anti-N-CAM antibodies inhibited the formation of histological layers in the retina. The data on perturbation by antibodies were correlated with the relative levels of expression of Ng-CAM and N-CAM in each of these different neural regions. Quantitative immunoblotting experiments indicated that the relative Ng-CAM/N-CAM ratios in comparable extracts of brain, dorsal root ganglia, and retina were respectively 0.32, 0.81, and 0.04. During culture of dorsal root ganglia in the presence of nerve growth factor, the Ng-CAM/N-CAM ratio rose to 4.95 in neurite outgrowths and 1.99 in the ganglion proper, reflecting both polarity and prevalence modulation. These results suggest that the relative ability of anti-Ng-CAM and anti-N-CAM antibodies to inhibit cell-cell interactions in different neural tissues is strongly correlated with the local Ng-CAM/N-CAM ratio.(ABSTRACT TRUNCATED AT 400 WORDS)



Heterotypic binding between neuronal membrane vesicles and glial cells is mediated by a specific cell adhesion molecule  

PubMed Central

By means of a multistage quantitative assay, we have identified a new kind of cell adhesion molecule (CAM) on neuronal cells of the chick embryo that is involved in their adhesion to glial cells. The assay used to identify the binding component (which we name neuron-glia CAM or Ng-CAM) was designed to distinguish between homotypic binding (e.g., neuron to neuron) and heterotypic binding (e.g., neuron to glia). This distinction was essential because a single neuron might simultaneously carry different CAMs separately mediating each of these interactions. The adhesion of neuronal cells to glial cells in vitro was previously found to be inhibited by Fab' fragments prepared from antisera against neuronal membranes but not by Fab' fragments against N-CAM, the neural cell adhesion molecule. This suggested that neuron-glia adhesion is mediated by specific cell surface molecules different from previously isolated CAMs . To verify that this was the case, neuronal membrane vesicles were labeled internally with 6-carboxyfluorescein and externally with 125I-labeled antibodies to N-CAM to block their homotypic binding. Labeled vesicles bound to glial cells but not to fibroblasts during a 30-min incubation period. The specific binding of the neuronal vesicles to glial cells was measured by fluorescence microscopy and gamma spectroscopy of the 125I label. Binding increased with increasing concentrations of both glial cells and neuronal vesicles. Fab' fragments prepared from anti-neuronal membrane sera that inhibited binding between neurons and glial cells were also found to inhibit neuronal vesicle binding to glial cells. The inhibitory activity of the Fab' fragments was depleted by preincubation with neuronal cells but not with glial cells. Trypsin treatment of neuronal membrane vesicles released material that neutralized Fab' fragment inhibition; after chromatography, neutralizing activity was enriched 50- fold. This fraction was injected into mice to produce monoclonal antibodies; an antibody was obtained that interacted with neurons, inhibited binding of neuronal membrane vesicles to glial cells, and recognized an Mr = 135,000 band in immunoblots of embryonic chick brain membranes. These results suggest that this molecule is present on the surfaces of neurons and that it directly or indirectly mediates adhesion between neurons and glial cells. Because the monoclonal antibody as well as the original polyspecific antibodies that were active in the assay did not bind to glial cells, we infer that neuron- glial interaction is heterophilic, i.e., it occurs between Ng-CAM on neurons and an as yet unidentified CAM present on glial cells.



Effects of prolonged oral administration of aflatoxin B1 and fumonisin B1 in broiler chickens.  


The effects of prolonged oral administration of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) mycotoxins were evaluated in broiler chickens from 21 to 42 d of age. A total of 192 birds were housed in experimental batteries and assigned to 32 cages, 6 birds per cage. The following treatments were applied: 1) 0 mycotoxins (control), 2) 10 mg of FB1, 3) 50 microg of AFB1, 4) 50 microg of AFB1 + 10 mg of FB1, 5) 350 microg of AFB1, 6) 350 microg of AFB1 + 10 mg of FB1, 7) 2,450 microg of AFB1, 8) 2,450 microg of AFB1 + 10 mg of FB1/kg of feed. Each treatment consisted of 4 replicates of 6 birds each. At the end of the trial, blood samples from 12 birds per treatment were collected, and the birds were necropsied. Compared with controls, the percentage of heterophils was lower (P < 0.05) in birds from groups receiving 50 microg of AFB1/kg + 10 mg of FB1/ kg and 2450 microg of AFB1/kg alone or in combination with FB1. A higher percentage of lymphocytes (P < 0.05) was observed in birds fed 50 microg of AFB1/kg + 10 mg of FB1/ kg, 350 microg of AFB1/kg, and 2,450 microg of AFB1/kg. A decrease in plasma albumin was observed only in birds fed 2,450 microg of AFB1/kg + 10 mg of FB1/kg. The liver of AFB1-treated birds had focal areas of necrosis and inflammatory infiltrates. In birds fed rations containing only 10 mg of FB1/kg, bile duct hyperplasia with fibrosis and a mononuclear infiltrate accompanied by trabecular derangement were observed. In contrast, in treatments in which FB1 was administered in combination, hepatic vacuolar degeneration was observed, and renal tissue presented corpuscles with increased cellular agglomeration, characterizing glomerulonephritis, and a clearly visible tubular epithelium with areas of degeneration and necrosis. The FB1 residues were detected in liver and in excreta of all FB1-treated groups, at levels that ranged from 0.013 to 0.051 mg/kg and from 1.19 to 2.79 mg/kg, respectively. Results indicated that FB1 and AFB1, singly or in combination at the levels evaluated, do not change markedly the hematological and serological parameters of broiler chickens, but may cause relevant lesions in liver and in kidneys. PMID:16479938

Del Bianchi, M; Oliveira, C A F; Albuquerque, R; Guerra, J L; Correa, B



The effects of water replacement by oral rehydration fluids with or without betaine supplementation on performance, acid-base balance, and water retention of heat-stressed broiler chickens.  


Exposing broilers to a high temperature increases water and electrolyte K(+) and Na(+) excretion, which negatively affects the heat dissipation capacity and acid-base homeostasis, resulting in losses in growth performance. In this experiment, the efficacy of providing oral rehydration therapy and betaine on growth performance, acid-base balance, and water and electrolyte retention was evaluated. A total of 432 one-day-old broiler chicks (Cobb) were allocated to 72 metabolic cages and reared to 31 d of age under standard conditions. From 32 to 41 d of age, chicks were exposed to heat stress (ambient temperature, 32°C) and high RH (80 to 100% RH) for 9 h daily. The ameliorative effects of a 3 × 3 factorial array of treatments administered via drinking water were evaluated in 8 replicates of 6 chicks per cage for each treatment. Two oral rehydration therapy (ORT) fluids, based on either citrate or bicarbonate salts, were added to tap water. In addition, betaine was added to tap water at an inclusion rate of 0, 500, or 1,000 mg/L to complete the array of 9 liquid-based treatments. Growth performance was assessed at 32, 35, and 41 d of age. From 32 to 35 d of age, chicks receiving ORT fluids exhibited improved growth performance, water balance, and electrolyte (K(+), Na(+)) retention. In addition, the physiological response to stress was attenuated, as indicated by lower heterophil-to-lymphocyte ratios and blood glucose concentrations relative to the negative controls. The addition of betaine at an inclusion rate of 500 mg/L improved BW gain. From d 36 to 41, treatments did not significantly influence growth performance, which suggests that chicks receiving tap water were able to compensate and adapt to the heat-stress conditions. The results demonstrate that the beneficial effects of providing ORT fluids and 500 mg of betaine/L were observed only during the first 4 d of heat exposure. After this period, adaptation to the heat appears to occur, and none of the treatments was successful in improving growth performance. PMID:21177455

Sayed, M A M; Downing, J



Effect of dietary ? -tocopherol concentration on performance and some immune responses in broiler chickens fed on diets containing oils from different sources.  


1. An investigation was carried out into the effects of dietary ?-tocopherol (?-T) concentration and source of supplemental oil on performance, activity of anti-oxidative enzymes and some immune responses in broilers from day-old to 41 d of age. 2. Three dietary concentrations of ?-T (10, 50 and 100 mg/kg) with three sources of supplemental oil (sunflower - SFO, palm - PMO and safflower - SAO) were provided using a 3 × 3 experimental design. 3. Body weight gain and food conversion efficiency were not affected by either interaction or concentrations of ?-T and sources of oil in diet. 4. Concentrations of total protein, globulin, triglycerides and cholesterol in sera increased significantly with dietary ?-T concentration irrespective of the source of oil. Significantly higher concentration of serum albumin was evident in broilers fed on the SFO-based diet and the concentration of globulin was higher in groups fed on those diets containing PMO and SAO. 5. The lipid peroxidation (LP), measured as MDA release, decreased with the concentration of ?-T in a dose-related manner with SFO- and SAO-based diets, although not with the PMO-based diet. With different oil sources, LP was significantly lower with the PMO-based diet compared to the others. Activities of glutathione peroxidase and RBC catalase increased and heterophil: lymphocyte ratio was reduced with concentration of ?-T for each source of oil tested. 6. Assays for humoral and cell-mediated immune responses indicated no effect of the source of dietary supplemental oil or interaction, although an increasing concentration of dietary ?-T improved cell-mediated immune responses. 7. It is concluded that sunflower oil, palm oil and safflower oil can be used as sources of oil for broiler diets without having any effect on performance, immune responses or the activity of anti-oxidizing enzymes. Higher concentrations of dietary ?-tocopherol (50 or 100 mg/kg) reduced lipid peroxidation activity and enhanced activities of anti-oxidative enzymes, they also improved the cell-mediated immune responses in commercial broilers. PMID:21337204

Rama Rao, S V; Raju, M V L N; Panda, A K; Poonam, N S; Shyam Sunder, G



Phosphacan, a chondroitin sulfate proteoglycan of brain that interacts with neurons and neural cell-adhesion molecules, is an extracellular variant of a receptor-type protein tyrosine phosphatase.  

PubMed Central

We have identified cDNA clones encoding a chondroitin sulfate proteoglycan of rat brain (previously designated 3F8 and now named phosphacan) that binds to neurons and neural cell-adhesion molecules. A sequence of 1616 amino acids deduced from a 4.8-kb open reading frame contains the N-terminal amino acid sequence of the 3F8 core glycoprotein as well as four internal CNBr, tryptic, and endoproteinase Lys-C peptide sequences from the proteoglycan. The deduced amino acid sequence, beginning with a 24-amino acid signal peptide, reveals an N-terminal domain of 255 amino acids homologous to carbonic anhydrases. The entire amino acid sequence deduced from our cDNA clones corresponds to the extracellular portion of a human receptor-type protein tyrosine phosphatase (RPTP zeta/beta) with which it has 76% identity, and the proteoglycan may represent an mRNA splicing variant of the larger transmembrane protein. RNA analysis demonstrated that a probe to the N-terminal carbonic anhydrase domain of the proteoglycan hybridizes with rat brain mRNA of 9.5, 8.4, and 6.4 kb, whereas probes to the phosphatase domains hybridize with only the 9.5-kb message and with the 6.4-kb message (which corresponds to a previously identified variant of the transmembrane protein in which half of the extracellular domain is deleted). The 30 N-terminal amino acids of the 3H1 chondroitin/keratan sulfate proteoglycan of brain are identical to those of the 3F8 proteoglycan, and six internal tryptic peptide sequences also matched those found in sequenced peptides of the 3F8 proteoglycan and/or amino acid sequences deduced from the cDNA clones. We therefore conclude that the 3H1 chondroitin/keratan sulfate proteoglycan and the 3F8 chondroitin sulfate proteoglycan represent glycosylation and possible extracellular splicing variants of a receptor-type protein tyrosine phosphatase. These proteoglycans may modulate cell interactions and other developmental processes in nervous tissue through heterophilic binding to cell-surface and extracellular matrix molecules, and by competition with ligands of the transmembrane phosphatase. Images

Maurel, P; Rauch, U; Flad, M; Margolis, R K; Margolis, R U



Effects of dietary selenium and vitamin E on immune response and biological blood parameters of broilers reared under thermoneutral or heat stress conditions.  


A study was conducted using 360 broiler chickens to evaluate the effects of dietary vitamin E (0, 125 and 250 mg/kg), selenium (Se, 0, 0.5 and 1 mg/kg), or their different combinations on immune response and blood biological parameters of broilers raised under either thermoneutral (TN, 23.9 °C constant) or heat stress (HS, 23.9 to 37 °C cycling) conditions. Humoral immunity was assessed by intravenous injection of 7 % sheep red blood cell (SRBC) followed by evaluation of serum for antibody titers in primary and secondary responses. Heterophil to lymphocyte (H/L) ratio also determined as an indicator of stress. Furthermore, at the end of the experiment, birds were bled for determination of some biological parameters. There was a significant reduction in body weight and feed intake, but the feed conversion ratio increased when the birds were exposed to HS (P??0.05), whereas feed conversion was improved significantly by 125 mg/kg vitamin E (P?

Habibian, Mahmood; Ghazi, Shahab; Moeini, Mohammad Mehdi; Abdolmohammadi, Alireza



Evaluation of the soft tissue biocompatibility of MgCa0.8 and surgical steel 316L in vivo: a comparative study in rabbits  

PubMed Central

Background Recent studies have shown the potential suitability of magnesium alloys as biodegradable implants. The aim of the present study was to compare the soft tissue biocompatibility of MgCa0.8 and commonly used surgical steel in vivo. Methods A biodegradable magnesium calcium alloy (MgCa0.8) and surgical steel (S316L), as a control, were investigated. Screws of identical geometrical conformation were implanted into the tibiae of 40 rabbits for a postoperative follow up of two, four, six and eight weeks. The tibialis cranialis muscle was in direct vicinity of the screw head and thus embedded in paraffin and histologically and immunohistochemically assessed. Haematoxylin and eosin staining was performed to identify macrophages, giant cells and heterophil granulocytes as well as the extent of tissue fibrosis and necrosis. Mouse anti-CD79? and rat anti-CD3 monoclonal primary antibodies were used for B- and T-lymphocyte detection. Evaluation of all sections was performed by applying a semi-quantitative score. Results Clinically, both implant materials were tolerated well. Histology revealed that a layer of fibrous tissue had formed between implant and overlying muscle in MgCa0.8 and S316L, which was demarcated by a layer of synoviocyte-like cells at its interface to the implant. In MgCa0.8 implants cavities were detected within the fibrous tissue, which were surrounded by the same kind of cell type. The thickness of the fibrous layer and the amount of tissue necrosis and cellular infiltrations gradually decreased in S316L. In contrast, a decrease could only be noted in the first weeks of implantation in MgCa0.8, whereas parameters were increasing again at the end of the observation period. B-lymphocytes were found more often in MgCa0.8 indicating humoral immunity and the presence of soluble antigens. Conversely, S316L displayed a higher quantity of T-lymphocytes. Conclusions Moderate inflammation was detected in both implant materials and resolved to a minimum during the first weeks indicating comparable biocompatibility for MgCa0.8 and S316L. Thus, the application of MgCa0.8 as biodegradable implant material seems conceivable. Since the inflammatory parameters were re-increasing at the end of the observation period in MgCa0.8 it is important to observe the development of inflammation over a longer time period in addition to the present study.



The L1-type cell adhesion molecule Neuroglian is necessary for maintenance of sensory axon advance in the Drosophila embryo  

PubMed Central

Background Cell adhesion molecules have long been implicated in the regulation of axon growth, but the precise cellular roles played by individual cell adhesion molecules and the molecular basis for their action are still not well understood. We have used the sensory system of the Drosophila embryo to shed light on the mechanism by which the L1-type cell adhesion molecule Neuroglian regulates axon growth. Results We have found a highly penetrant sensory axon stalling phenotype in neuroglian mutant embryos. Axons stalled at a variety of positions along their normal trajectory, but most commonly in the periphery some distance along the peripheral nerve. All lateral and dorsal cluster sensory neurons examined, except for the dorsal cluster neuron dbd, showed stalling. Sensory axons were never seen to project along inappropriate pathways in neuroglian mutants and stalled axons showed normal patterns of fasciculation within nerves. The growth cones of stalled axons possessed a simple morphology, similar to their appearance in wild-type embryos when advancing along nerves. Driving expression of the wild-type form of Neuroglian in sensory neurons alone rescued the neuroglian mutant phenotype of both pioneering and follower neurons. A partial rescue was achieved by expressing the Neuroglian extracellular domain. Over/mis-expression of Neuroglian in all neurons, oenocytes or trachea had no apparent effect on sensory axon growth. Conclusion We conclude that Neuroglian is necessary to maintain axon advance along axonal substrates, but is not required for initiation of axon outgrowth, axon fasciculation or recognition of correct growth substrates. Expression of Neuroglian in sensory neurons alone is sufficient to promote axon advance and the intracellular region of the molecule is largely dispensable for this function. It is unlikely, therefore, that Nrg acts as a molecular 'clutch' to couple adhesion of F-actin within the growth cone to the extracellular substrate. Rather, we suggest that Neuroglian mediates sensory axon advance by promoting adhesion of the surface of the growth cone to its substrate. Our finding that stalling of a pioneer sensory neuron is rescued by driving Neuroglian in sensory neurons alone may suggest that Neuroglian can act in a heterophilic fashion.

Martin, Veronica; Mrkusich, Eli; Steinel, Martin C; Rice, Jason; Merritt, David J; Whitington, Paul M



Bacteriological and pathological studies of egg peritonitis in commerciallayer chicken in Namakkal area  

PubMed Central

Objective To detect the various bacteriological agents and pathological changes in commercial layer chicken affected with egg yolk peritonitis in Namakkal region of India. Methods A total of 6?572 layer chicken from 85 commercial farms were subjected for the study, out of which 1?715 showed various types of oviduct abnormalities. Among the 1?715, 264 birds from six farms were identified as egg peritonitis on the basis of postmortem examination. Trachea, lung, heart blood, liver, peritoneal exudate, oviduct (infundibulum, magnum, uterus) and cloacal swabs were collected from the 264 birds with egg peritonitis lesion for screening of bacterial agents. Signalment, clinical signs and pathological changes were recorded in the affected flocks. Result The results of the present investigation indicated that the E. coli associated egg peritonitis was responsible for 15.39% of the reproductive tract abnormalities in commercial layers between 21 and 80 week of age. In the affected flocks egg production drop and mortality varied from 3% to 20% and 0.5% to 7.0% respectively. It was noticed during peak egg production (21 to 60 week) and southwest monsoon season (58%). Statistical analysis of age, season and egg production by Chi square test of independence revealed highly significant difference. E. coli was isolated as a pure culture and concurrent with other bacterial agents in 226 and 38 birds respectively. Among the fifteen E. coli serotypes identified serotype O166, O64 and O111 were predominant. Necropsy examination of affected birds revealed the presence of amorphous or insipissiated yolk material in the abdominal cavity with inflammatory changes in the ovary, oviduct and intestine. Microscopically the oviduct surface epithelium showed degeneration and desquamation, moderate to marked infiltration of inflammatory cells especially heterophils and lymphocytes in various regions and lumen contained serofibrinous exudate, inflammatory and desquamated epithelial cells with bacterial microcolonies. Ovarian follicles revealed hyperemia, degeneration of granulosa cells and infiltration of inflammatory cells. Intestine showed degenerative, necrotic and inflammatory lesion. Conclusion The findings of this study showed that the egg peritonitis might be caused by either the translocation of intestinal E. coli into the peritoneal cavity or by the movement of cloacal E. coli into the oviduct followed by ascension of these bacteria up the oviduct, through the infundibulum, and into the peritoneal cavity. To control the egg peritonitis faecal contamination with E. coli should be minimized.

Srinivasan, Palani; Balasubramaniam, Gurusamypalayam Amirthalingam; Murthy, Thippichettipalayam Ramasamy Gopala Krishna; Balachandran, Perumal



Testing a West Nile virus vaccine in sandhill cranes (Grus canadensis)  

USGS Publications Warehouse

Eight sandhill cranes (Grus canadensis) were vaccinated with a commercial equine West Nile virus vaccine (Fort Dodge Animal Health, Fort Dodge, Iowa, USA) at the USGS Patuxent Wildlife Research Center, Laurel, Maryland, USA. Three doses of the vaccine were given, the first dose (day 0) was followed by a second 21 days later and the third dose 7 days after the second day 28 after the first dose). All doses were 0.50 ml. In addition, 5 sandhill cranes were given injections of similar amounts of sterile water on the same schedule. Blood for complete blood counts, serum chemistries, and serological testing was collected at weekly intervals. Ten weeks after the first injection of the vaccine and 6 weeks after the last of the 3 injections of the vaccine, the cranes were shipped to the USGS National Wildlife Heath Center, Madison, Wisconsin, USA. After a two-week adjustment period, 11 of these cranes were injected intramuscularly with one mosquito dose of West Nile virus. Two of the vaccinated cranes were not challenged and acted as uninfected controls. One week post challenge the only abnormal findings were slight weight loss (average 6% loss since the time of challenge with West Nile virus) and elevated white blood cell counts (heterophilic leukocytosis). There were no deaths and no clinically ill cranes (unvaccinated or vaccinated cranes) among the 11 challenged birds. All cranes were euthanized 6 weeks post challenge, and necropsies were performed. Pre-challenge titers showed no titer response to the vaccinations. However, when challenged, vaccinated cranes developed titers more quickly (within 7-10 days), and were viremic and shed virus via the cloaca for a shorter period of time than the unvaccinated cranes (2-7 days for vaccinated cranes versus 2-10 days for unvaccinated cranes). No remarkable lesions were noted in any of the cranes during the necropsy examinations. Histopathological findings are available for only four of the cranes at this time. Three of those had mild to moderate encephalitis, myelitis, and myocarditis. Initial histopathological findings also indicated a difference in the lesions found in the brains of the vaccinated versus unvaccinated cranes. The fourth crane was a vaccinated unchallenged control that had none of the lesions described. We concluded, based on the findings of no adverse reactions and the higher titers and reduced viremia seen in the cranes, that, indeed, the vaccine is safe for use in cranes and can be effective in reducing the severity of a natural infection. We would recommend this vaccine for use in adult cranes. A companion study is looking at the safety and efficacy of the vaccine for crane chicks as young as 7-10 days of age and that will be reported at a later date.

Olsen, G.H.; Miller, K.; Docherty, D.; Sileo, L.