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Heterophile antibodies in Nigerian sera  

PubMed Central

A heterophile agglutinin was found at a titre of 1:4 or greater in 332 of 336 Nigerian sera investigated. The antibody was demonstrated to be an IgM macroglobulin. Although many of the sera tested had high IgM levels, only a slight correlation was found between titres of heterophile agglutinin and IgM levels. Absorption studies differentiated the Nigerian heterophile agglutinin from the antibodies seen in glandular fever and serum sickness. No correlation was found between the occurrence of high titres of heterophile agglutinin and infection with malaria, onchocerciasis, loaisis or schistosomiasis. None of the subjects investigated was known to have trypanosomiasis, a parasitic infection in which heterophile antibodies are known to occur.

Greenwood, B. M.



Heterophil cytokine mRNA profiles from genetically distinct lines of chickens with differential heterophil-mediated innate immune responses  

Microsoft Academic Search

Previously we demonstrated that increased in-vitro heterophil function translates to increased in-vivo resistance to Salmonella enteritidis infections in broilers (line A?>?B). Heterophils produce cytokines and modulate acute protection against Salmonella in neonatal poultry. We hypothesized that heterophils from S. enteritidis-resistant chickens produce an up-regulated pro-inflammatory cytokine\\/chemokine response compared with S. enteritidis-susceptible chickens. In this study, heterophils were isolated 1, 14,

Christina L. Swaggerty; Pete Kaiser; Lisa Rothwell; Igal Y. Pevzner; Michael H. Kogut



Pseudohypergonadotropinemia and pseudohyperprolactinemia induced by heterophilic antibodies?  


The serum of 20 apparently hypergonadotropic and/or hyperprolactinemic patients (14 females, 6 males, ages 13-75 years) without evidence of neoplasia or pituitary adenomas was found to contain a large amount of molecular material (MW ca. 100,000) resulting in factitiously elevated levels of peptide hormones when measured by double-antibody radioimmunoassay with a long second incubation time. The interference by this material with the test system could be avoided by using polyethylene glycol (PEG) for the separation of free from bound antigen, or by preincubation of the samples with normal rabbit serum. No definite disease process can as yet be linked to these findings. They rather seem to be caused by the presence of heterophilic antibodies in serum, as the serum of approximately one half of the patients was found to give a positive Paul-Bunnell test. Moreover, the beta hCG activity in the urine of these patients was low, probably as a result of the low clearance of substances with igh MW. It is therefore suggested to apply to following diagnostic measures before clinical consequences are being considered in a case of hypergonadotropinemia and/or hyperprolactinemia without pertinent clinical findings: (1) validation of the immunoassay, (2) preincubation of the samples with serum from other species, (3) the use of another separation procedure than the double-antibody method, and (4) measurement of the respective hormone in urine. PMID:6425554

Dericks-Tan, J S; Jost, A; Schwedes, U; Taubert, H D




Technology Transfer Automated Retrieval System (TEKTRAN)

A fluorescence microplate assay was adapted to examine the oxidative response by heterophils from neonatal chicks following in vitro stimulation with various inflammatory agonists. Both nonopsonized formalin-killed Salmonella enteritidis and Staphylococcus aureus stimulated significant heterophil o...



Technology Transfer Automated Retrieval System (TEKTRAN)

Synthetic oligodeoxynucleotide containing unmethylated CpG motif (CpG-ODN) is immune stimulatory to chicken heterophils. Recognition of CpG-ODN by chicken heterophils leads to the mobilization and release of granules. This CpG-ODN-induced heterophil degranulation was chicken serum (CS)-dependent. ...



Technology Transfer Automated Retrieval System (TEKTRAN)

Synthetic oligodeoxynucleotide containing unmethylated CpG motif (CpG-ODN) is stimulatory to chicken heterophils. Recognition of CpG-ODN by chicken heterophils leads to the mobilization and release of granules. This CpG-ODN-induced heterophil degranulation was serum-dependent. Removal of chicken ...


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.




Technology Transfer Automated Retrieval System (TEKTRAN)

We reported differences in in vitro heterophil functional efficiency between parental lines (A>B) of broilers and the F1 reciprocal crosses (D>C). Heterophils modulate acute protection against Salmonella in young poultry. Therefore, we are interested in evaluating susceptibility of the four lines ...


In vivo activation of heterophil function in chickens following injection with Salmonella enteritidis-immune lymphokines  

Microsoft Academic Search

We have previously shown that increased re- sistance to Salmonella enteritidis organ infectivity in day-old chicks was conferred by the immunoprophylactic administration of S. enteritidis-immune lymphokines (ILK). This resistance was associated with a significant increase in the number of circulating heterophils 4 h after ILK injection. The objective of the present study was to evaluate heterophil function following the administration

Michael H. Kogut; Edward D. McGruder; Billy M. Hargis; Donald E. Corrier; John A. DeLoach




Technology Transfer Automated Retrieval System (TEKTRAN)

We recently showed that in vitro heterophil functional efficiency in commercial broiler chickens is genetically controlled and may be a sex-associated trait. To further characterize the genetic mechanism(s) of heterophil functional efficiency we wanted to determine if the feathering gene, present ...



PubMed Central

For ultrastructural localization of acid mucosubstances in rabbit granulocytes, bone marrow and buffy coat specimens were fixed with formalin, glutaraldehyde, or osmium tetroxide, sectioned at 40 µ, and stained with the Rinehart and Abul-Haj solution of dialyzed iron (DI). Heterophils revealed DI staining on the outer surface of the plasma membrane, in the Golgi complex involved in primary granulogenesis, and in primary granules. The intragranular distribution of DI-stained material varied at different stages in the maturation of primary granules. Immature granules of heterophils fixed by any of the three methods contained a peripheral concentric band of DI-positive material; however, fully mature primary granules possessed a core of DI-reactive material in heterophils fixed with osmium tetroxide, but they contained little or no staining in heterophils fixed with formalin or glutaraldehyde. Secondary granules of rabbit heterophils failed to stain with DI. Tertiary granules, observed only in late heterophils, contained distinct DI-positive particles. Basophil granules exhibited intensely DI-stained material distributed in an orderly pattern throughout the granule. In eosinophils, DI staining was localized in the Golgi complex and in the rims of a few immature cytoplasmic granules.

Hardin, J. H.; Spicer, S. S.



Heterophilic antibodies interfering with radioimmunoassay. A false-positive pregnancy test  

SciTech Connect

A young woman with amenorrhea had a consistently positive pregnancy test result (serum radioimmunoassay measurement of ..beta..-human chorionic gonadotropin hormone). No fetal or placental tissue was found after uterine curettage and exploratory laparotomy. The false-positive pregnancy test result was due to heterophilic antibovine and antigoat antibodies in the patient's serum. These antibodies interfered with radioimmunoassays using goat antibodies. This case shows that serum heterophilic antibodies can interfere with immunoassays and result in unnecessary diagnostic procedures and/or unnecessary treatment.

Vladutiu, A.O.; Sulewski, J.M.; Pudlak, K.A.; Stull, C.G.




Microsoft Academic Search

We measured corticosterone in plasma collected from free-living AdClie Pen- guins (Pygoscelis adeliae) and made blood smears to count the ratio of heterophils to lym- phocytes (Ha). Our objective was to categorize how these two measures of stress vary with potential stressors in the birds' environment. In penguins that were repeatedly sampled from three to eight times, repeated handling did

Carol M. Vleck; Nicole Vertalino; David Vleck; Theresa L. Bucher



Direct screening identifies mature beta-defensin 2 in avian heterophils  

Technology Transfer Automated Retrieval System (TEKTRAN)

Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used to screen avian heterophils in the m/z range of 1-20 kDa with an objective to identify the cell associated peptides that may be reflective of their functional physiology. The MALDI-TOF-MS profiles ...



Technology Transfer Automated Retrieval System (TEKTRAN)

A comparison of cell signaling and functions in heterophils from a commercial line (A) to wild-type Rio Grande turkeys days 4 and 7 post-hatch was conducted. Mitogen-activated protein kinases (MAPK) p38 and ERK 1/2 and total protein tyrosine kinase (PTK) activities were measured. After stimulation...


Raised tryptase without anaphylaxis or mastocytosis: heterophilic antibody interference in the serum tryptase assay.  


Mast cell tryptase (MCT) is a key diagnostic test for mastocytosis and anaphylaxis. High serum tryptase levels are also one of the risk factors for adverse reaction in venom immunotherapy, yet occasional patients are seen with raised levels in the absence of either diagnosis. False positive results can be due to assay interference by heterophilic antibodies such as rheumatoid factor (RF) and human anti-mouse antibodies (HAMA). We therefore investigated heterophilic antibody interference by rheumatoid factor activity and HAMA as a cause of raised MCT results in the Phadia tryptase assay. Serum samples from 83 patients were assayed for MCT and rheumatoid factor before and after the use of heterophilic antibody blocking tubes (HBT). Samples with more than 17% reduction in MCT with detectable RF were then assayed for HAMA. Fourteen (17%) of the 83 samples with positive RF showed a >17% decrease in mast cell tryptase after HBT blocking. Post-HBT, eight of 14 (57%) reverted from elevated to normal range values with falls of up to 98%. RF levels were also decreased significantly (up to 75%). Only one of the 83 tested was apparently affected by HAMA in the absence of detectable IgM RF. In conclusion, any suspicious MCT result should be checked for heterophilic antibodies to evaluate possible interference. False positive MCT levels can be caused by rheumatoid factor. We suggest a strategy for identifying assay interference, and show that it is essential to incorporate this caveat into guidance for interpretation of MCT results. PMID:21303361

Sargur, R; Cowley, D; Murng, S; Wild, G; Green, K; Shrimpton, A; Egner, W



Heterophile Antibody Interference in a Multiplexed Fluorescent Microsphere Immunoassay for Quantitation of Cytokines in Human Serum  

Microsoft Academic Search

While modern immunoassays provide sensitive and specific means for the quantitation of cytokines in biological fluids, heterophile antibodies are still a well-recognized cause of interference in the measurement of cytokines in these assays. We have developed a multiplexed fluorescent microsphere immunoassay for the simultaneous quantification of 10 cytokines in only 75 l of serum. During the development of this multiplexed

Thomas B Martins; Brian M. Pasi; Christine M. Litwin; Harry R. Hill



BT cationic peptides: small peptides that modulate innate immune responses of chicken heterophils and monocytes.  


Neonatal poultry exhibit a transient susceptibility to infectious diseases during the first week of life that stems from inefficient host defense mechanisms. Yet, the initial host immune response to pathogens is a critical determinant of disease resistance and susceptibility. With this context in mind, novel ways to stimulate or modulate the hosts' natural immune response is emerging as an important area of interest for food animal producers including the poultry industry. Specifically, we have been investigating new modulation strategies tailored around the selective stimulation of the host's immune system, and particularly rapid acting innate immunity, as an alternative to direct targeting of microbial pathogens. One such approach that we have been investigating is the use of a group of cationic peptides produced by a Gram-positive soil bacterium, Brevibacillus texasporus (BT peptides). We have previously shown that, provided as a feed additive, BT peptides significantly induced a concentration-dependent protection against cecal colonization and extraintestinal colonization by Salmonella enterica serovar Enteritidis (SE). This protection is not the result of direct antibacterial activity of the BT peptides on the SE since the concentrations used were below the minimum inhibitory concentration for SE. We also found that BT are not absorbed in the intestine, but still induce a significant up-regulation in the functional efficiency of peripheral blood heterophils and monocytes. The mechanisms of this immune modulation are unknown. Here, using in vitro models for measuring: (1) leukocyte oxidative burst, (2) changes in leukocyte cytokine and chemokines gene expression profiles, and (3) phosphorylation of the mitogen activated protein kinases (MAPKs) in leukocytes, we evaluated the role of BT peptides as priming mediators for heterophil and monocyte responses at the level of cell function, gene transcription/expression, and cell phosphorylation following stimulation with inflammatory agonists. BT peptides primed both heterophils and monocytes for an increased oxidative burst and up-regulation in transcription of the pro-inflammatory cytokines IL-1? and IL-6 and inflammatory chemokines CXCLi1 and CXCLi2 induced by inflammatory agonists. In addition, BT peptides induced a rapid (10min) phosphorylation and activation of the extracellular signal-regulated kinase (ERK1/2) and p38 kinase pathways in primary chicken heterophils. Taken together, we conclude that BT peptides, acting through MAPK pathways, enhance leukocyte functional and pro-inflammatory cytokine and chemokine gene transcription activities. These small cationic peptides may prove useful as immune modulators in neonatal poultry. PMID:22129785

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




Technology Transfer Automated Retrieval System (TEKTRAN)

Toll-like receptors (TLRs) have been previously shown to mediate oxidative burst in chicken heterophils. This study was conducted to determine which molecular pathways are involved in TLR mediated oxidative burst. Peripheral blood heterophils from neonatal chicks were isolated and exposed to known...



Technology Transfer Automated Retrieval System (TEKTRAN)

We have previously reported the inhibition of Fc receptor-mediated degranulation in avian heterophils by the syk tyrosine kinase inhibitor piceatannol. The present studies investigated whether attachment of complement opsonized bacteria to complement receptors also involve the syk tyrosine kinase p...



Technology Transfer Automated Retrieval System (TEKTRAN)

We have conducted studies utilizing a parental pair of broiler chickens (lines A and B) and the F1 reciprocal crosses (C and D). Previously we showed increased in vitro heterophil functional efficiency translates to increased in vivo resistance against a systemic Salmonella enteritidis (SE) infecti...


Interference by heterophilic antibodies in immunoassays: wrong increase of myoglobin values.  


Aim of this work is to illustrate how analytical interference in immunoassay may produce serious errors in clinical laboratory results. The sophisticated quality assurance schemes used in many laboratories do not identify erroneous results arising from aberrant samples. Recently attention has been focused on the incidence and implication of false-positive results arising from the presence of certain substances in a patient's serum that interfere with one or more steps in immunoassays. In this paper, we present the case of a 92 year-old woman whose plasma myoglobin concentrations falsely increased when measured using the Beckman Access assay. We demonstrated that heterophilic antibodies accounted for the falsely increased myoglobin values, and we suggest how to resolve such situations. PMID:18788511

Bonetti, Antonio; Monica, Cesare; Bonaguri, Chiara; Gnocchi, Cecilia; Russo, Annalisa; Battistelli, Luisita; Musiari, Luisa; Pastori, Paolo; Novarini, Almerico



Studies on some heterophile receptors of the Burkitt EB2 lymphoma cell  

PubMed Central

Using monospecific reagents from invertebrates, particularly those directed against terminal N-acetyl-neuraminyl residues, it is deduced that Burkitt EB2 lymphoma cells have terminal N-acetyl-neuraminyl groups linked glycosidically to D-galactose units. Evidence was obtained with agglutinins derived from plants that terminal D-galactosyl and inner-chain N-acetyl-glucosaminyl groups are also components of heterosaccharide chains in the cell membrane. Neither terminal nor penultimate N-acetyl-D-galactosaminyl groups were detected by N-acetyl-D-galactosaminyl specific snail agglutinins, nor were myxovirus receptors present to any significant extent, thus differentiating Burkitt EB2 lymphoma cells from erythrocytes. The T- and Forssman antigens were absent. Burkitt EB2 lymphoma cells actively regenerate neuraminyl end-groups following treatment with neuraminidase; this biosynthesis is suppressed by sub-lethal irradiation. Several other heterophile lectin receptors were detected in Burkitt EB2 lymphoma cells during our investigations, and it was confirmed that Phaseolus vulgaris (PHA) extract has separate erythrocyte-agglutinating and mitogenic properties. All studies were carried out on intact living Burkitt EB2 lymphoma cells.

Pardoe, G. I.; Uhlenbruck, G.; Bird, G. W. G.



Gene expression profiling in chicken heterophils with Salmonella enteritidis stimulation using a chicken 44 K Agilent microarray  

PubMed Central

Background Salmonella enterica serovar Enteritidis (SE) is one of the most common food-borne pathogens that cause human salmonellosis and usually results from the consumption of contaminated poultry products. The mechanism of SE resistance in chickens remains largely unknown. Previously, heterophils isolated from broilers with different genetic backgrounds (SE-resistant [line A] and -susceptible [line B]) have been shown to be important in defending against SE infections. To dissect the interplay between heterophils and SE infection, we utilized large-scale gene expression profiling. Results The results showed more differentially expressed genes were found between different lines than between infection (SE-treated) and non-infection (control) samples within line. However, the numbers of expressed immune-related genes between these two comparisons were dramatically different. More genes related to immune function were down-regulated in line B than line A. The analysis of the immune-related genes indicated that SE infection induced a stronger, up-regulated gene expression of line heterophils A than line B, and these genes include several components in the Toll-like receptor (TLR) signaling pathway, and genes involved in T-helper cell activation. Conclusion We found: (1) A divergent expression pattern of immune-related genes between lines of different genetic backgrounds. The higher expression of immune-related genes might be more beneficial to enhance host immunity in the resistant line; (2) a similar TLR regulatory network might exist in both lines, where a possible MyD88-independent pathway may participate in the regulation of host innate immunity; (3) the genes exclusively differentially expressed in line A or line B with SE infection provided strong candidates for further investigating SE resistance and susceptibility. These findings have laid the foundation for future studies of TLR pathway regulation and cellular modulation of SE infection in chickens.

Chiang, Hsin-I; Swaggerty, Christina L; Kogut, Michael H; Dowd, Scot E; Li, Xianyao; Pevzner, Igal Y; Zhou, Huaijun



Impact of age and heterophilic interference on the basal serum tryptase, a risk indication for anaphylaxis, in 1,092 dermatology patients.  


A raised baseline serum tryptase is a risk indicator for anaphylactic reactions, especially in patients with hymenoptera venom allergy. Borderline elevations (> 11.4 ?g/l) occur frequently and may necessitate invasive diagnostic procedures to rule out systemic mastocytosis. We retrospectively analysed 1,092 non-mastocytotic patients from our general dermatology clinic with respect to age- and gender-associated effects and investigated the impact of heterophilic antibody interference on the tryptase assay. The results were stratified by gender and five age classes. Sera with raised tryptase (n = 106) were re-tested after pre-incubation with Heterophilic Blocking Tubes (HBT(®), Scantibodies Laboratory; Santee, CA, USA). A significant increase in baseline tryptase was observed with increasing age. Incubation with HBT(®) caused a decline of more than 50% in only one case. In conclusion, older patients showed significantly higher serum tryptase levels and heterophilic interference was of subordinate relevance. PMID:22170044

Schliemann, Sibylle; Seyfarth, Florian; Hipler, Uta-Christina; Elsner, Peter



The macrophage inflammatory protein-1beta in the supernatants of Mycoplasma gallisepticum-infected chicken leukocytes attracts the migration of chicken heterophils and lymphocytes.  


Chicken monocytes, macrophages, heterophils and thrombocytes were infected with Mycoplasma gallisepticum, and their supernatants were collected and tested for the presence of chemotactic activities. The supernatants from MG-infected monocytes and macrophages were able to attract the migration of both heterophils and lymphocytes. The chemotactic activity in these supernatants could be abolished by antibodies prepared against the 10 amino acid peptides of the macrophage inflammatory protein (MIP)-1beta, indicating that the released chemoattractant was a MIP-1beta-like compound. The supernatant from MG-infected heterophils was also able to attract the migration of chicken lymphocytes, but its activity could not be neutralized by the antibody to MIP-1beta, indicating that the chemoattractant is not related to MIP-1beta. The supernatants from both control and MG-infected thrombocytes were able to attract the migration of lymphocytes. These results indicate that there is more than one chemotactic factor that is released by these cells; one of the chemoattractants has been identified as a MIP-1beta. These results also show that MIP-1beta may play a role in the recruitment and accumulation of heterophils and lymphocytes to the sites of mycoplasma infection. PMID:11687266

Lam, K M



In vivo priming heterophil innate immune functions and increasing resistance to Salmonella enteritidis infection in neonatal chickens by immune stimulatory CpG-ODN  

Technology Transfer Automated Retrieval System (TEKTRAN)

Oligodeoxynucleotides (ODN) containing CpG dinucleotides (CpG-ODN) mimic bacterial DNA and stimulate the innate immune system of vertebrates. Here, we investigated the effects of intraperitoneal (ip) administered CpG-ODN on the innate immune functions of chicken heterophils. Our results demonstrat...


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



Selective pharmacological inhibitors reveal the role of Syk tyrosine kinase, phospholipase C, phosphatidylinositol-3?-kinase, and p38 mitogen-activated protein kinase in Fc receptor-mediated signaling of chicken heterophil degranulation  

Microsoft Academic Search

Fc receptors of avian heterophils play a primary role in the elimination of bacterial pathogens in poultry. The cross-linking of Fc receptors with IgG-bacteria complexes results in the secretion of toxic oxygen metabolites and anti-bacterial granules. We have been investigating the upstream signaling events that precede degranulation following crosslinkage of Fc receptors on heterophils. Previously when using the non-selective pharmacological

Michael Kogut; Virginia K. Lowry; Morgan Farnell



The use of selective pharmacological inhibitors to delineate signal transduction pathways activated during complement receptor-mediated degranulation in chicken heterophils  

Microsoft Academic Search

Complement receptors (CRs), along with Fc receptors, play a primary role in the removal of bacterial pathogens in poultry. The binding of serum-opsonized bacteria to CR results in the secretion of both toxic oxygen metabolites and antibacterial granules. We have previously shown that the stimulation of chicken heterophils with serum-opsonized Salmonella enteritidis induced tyrosine kinase-dependent phosphorylation regulated degranulation. In the

Michael H. Kogut; Virginia K. Lowry; Morgan Farnell



Differential activation of signal transduction pathways mediating phagocytosis, oxidative burst, and degranulation by chicken heterophils in response to stimulation with opsonized Salmonella enteritidis.  


The activation of signal transduction pathways is required for the expression of functional enhancement of cellular activities. In the present studies, initial attempts were made to identify the signal transduction factors involved in activating phagocytosis, generation of an oxidative burst, and degranulation by heterophils isolated from neonatal chickens in response to opsonized Salmonella enteritidis (opsonized SE). Peripheral blood heterophils were isolated and exposed to known inhibitors of signal transduction pathways for either 20 min (staurosporin, genistein, or verapamil) or 120 min (pertussis toxin) at 39 degrees C. The cells were then stimulated for 30 min at 39 degrees C with opsonized SE. Phagocytosis, luminol-dependent chemoluminescence (LDCL), and beta-D glucuronidase release were then evaluated in vitro. The G-protein inhibitor pertussin toxin markedly inhibited (>80%) phagocytosis of opsonized SE. Both the protein kinase inhibitor (staurosporin) and calcium channel inhibitor (verapamil) reduced phagocytosis in a dose response manner. Genistein, a tyrosine kinase inhibitor, had no effect on phagocytosis. Staurosporin had a marked inhibitory effect on LDCL (>90%) while genistein had a dose responsive inhibition on LDCL. Both verapamil (40-45%) and pertussin toxin (50-55%) had a statistically significant, but less biologically significant effect on LDCL. Genistein significantly reduced the degranulation (78-81%) of heterophils by opsonized SE. Staurosporin also reduced degranulation by 43-50%, but neither verapamil nor pertussis toxin had a significant effect on degranulation. These findings demonstrate that distinct signal transduction pathways differentially regulate the stimulation of the functional activities of avian heterophils. Pertussin toxin-sensitive, Ca++-dependent G-proteins appear to regulate phagocytosis of opsonized SE, protein kinase C-dependent, tyrosine kinase-dependent protein phosphorylation plays a major role in LDCL, and tyrosine kinase(s)-dependent phosphorylation regulates primary granule release. PMID:11293667

Kogut, M H; Genovese, K J; Lowry, V K



Fluorescein isothiocyanate staining and characterization of avian heterophils 1 Mention of a trade name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the USDA and does not imply its approval to the exclusion of other products that may be suitable. 1  

Microsoft Academic Search

Fluorescein isothiocyanate (FITC) was found to stain cytoplasmic granules of avian heterophil-granulocytes. In tissue sections, the fluorescent granulocytes were predominantly distributed adjacent to trabecular bones. The fluorescein stained granulocytes were abundant in synovial fluids of chickens with synovitis. A significant correlation was observed in the percent of fluorescein labeled granulocytes in blood smears and the percent of heterophils determined using

N. C Rath; G. R Huff; J. M Balog; W. E Huff



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



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


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. PMID:24066123

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



Heterophilic antigens and antibodies in fresh water and marine fish  

PubMed Central

Reciprocal agglutination studies using reagents from six fresh water fish species indicated distinct patterns between the behaviour of carp, bullhead and pickerel erythrocytes, and those derived from bass and from sunfish in contrast to perch erythrocytes, which were unreactive in the presence of sera from other fish species. Each of these could be further differentiated from one another by reciprocal absorption experiments or by the use of unabsorbed or absorbed serum from human group O, A, B and AB individuals. In similar fashion, it could be shown that members of three marine fish species differed from one another in regard to agglutinogen content. These were the Conger eel, red hind and the toadfish. Absorption experiments also demonstrated the presence of a factor or factors common to the latter species, but absent in the brown bullhead.

Kuhns, W. J.; Chuba, J. V.; Nigrelli, R. F.



Bacterial clearance, heterophil function, and hematological parameters of transport stressed turkey poults supplemented with dietary yeast extract  

Technology Transfer Automated Retrieval System (TEKTRAN)

Yeast extracts contain biological response modifiers that may be useful as alternatives to antibiotics for controlling pathogens in poultry production and mitigating the deleterious effects of production stressors. A standardized yeast extract feed supplement, Alphamune™ (YE), was added to turkey po...


Direct Ca2+-dependent heterophilic interaction between desmosomal cadherins, desmoglein and desmocollin, contributes to cell-cell adhesion.  


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. PMID:9214392

Chitaev, N A; Troyanovsky, S M



Effects of a dietary yeast extract on hematological parameters, heterophil function, and bacterial clearance in turkey poults challenged with Escherichia coli and subjected to transport stress  

Technology Transfer Automated Retrieval System (TEKTRAN)

There is a need to develop nutritional methods for controlling pathogens in poultry production. A standardized yeast extract supplement, Alphamune™ (YE), was added to turkey poult diets. Male poults were challenged by air sac injection with 60 cfu of E. coli at 1 week of age. At 3 weeks of age chal...


The ancillary protein 1 of Streptococcus pyogenes FCT-1 pili mediates cell adhesion and biofilm formation through heterophilic as well as homophilic interactions  

PubMed Central

Summary Gram-positive pili are known to play a role in bacterial adhesion to epithelial cells and in the formation of biofilm microbial communities. In the present study we undertook the functional characterization of the pilus ancillary protein 1 (AP1_M6) from Streptococcus pyogenes isolates expressing the FCT-1 pilus variant, known to be strong biofilm formers. Cell binding and biofilm formation assays using S. pyogenes in-frame deletion mutants, Lactococcus expressing heterologous FCT-1 pili and purified recombinant AP1_M6, indicated that this pilin is a strong cell adhesin that is also involved in bacterial biofilm formation. Moreover, we show that AP1_M6 establishes homophilic interactions that mediate inter-bacterial contact, possibly promoting bacterial colonization of target epithelial cells in the form of three-dimensional microcolonies. Finally, AP1_M6 knockout mutants were less virulent in mice, indicating that this protein is also implicated in GAS systemic infection.

Becherelli, Marco; Manetti, Andrea G O; Buccato, Scilla; Viciani, Elisa; Ciucchi, Laura; Mollica, Giulia; Grandi, Guido; Margarit, Imma



Differential induction of nitric oxide, degranulation, and oxidative burst activities in response to microbial agonist stimulation in monocytes and heterophils from young commercial turkeys  

Technology Transfer Automated Retrieval System (TEKTRAN)

The Toll-like receptors (TLRs) recognize microbial pathogens and pathogen associated molecular patterns and trigger inflammatory immune responses to control the infection. Here, we examined functional innate immune responses to Salmonella enteritidis (SE, live or formalin-killed) and various TLR ag...



Technology Transfer Automated Retrieval System (TEKTRAN)

Complement receptors (CR), along with Fc receptors, play a primary role in the removal of bacterial pathogens in poultry. The binding of serum-opsonized bacteria to CR results in the secretion of both toxic oxygen metabolites and anti-bacterial granules. We have previously shown that the stimulati...


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 ...


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



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




Technology Transfer Automated Retrieval System (TEKTRAN)

The immediate response to invasive pathogens, clearance via the inflammatory response, and activation of appropriate acquired responses are all coordinated by innate host defenses. Recognition of microbes is accompanied by the induction of multiple cellular processes including the production of pro...


Patterns of variation in leucocyte counts of female tree swallows, Tachycineta bicolor: Repeatability over time and relationships with condition and costs of reproduction  

Microsoft Academic Search

The heterophil to lymphocyte (HL) ratio in the peripheral blood is increasingly being recognized as a reliable indicator of stress in birds. In this study we examined whether HL ratio, as well as the proportion of heterophils and lymphocytes, varied systematically with sampling time, date, age, climate conditions or with measures of maternal investment in female tree swallows (Tachycineta bicolor)

Courtney L. Ochs; Russell D. Dawson




Technology Transfer Automated Retrieval System (TEKTRAN)

In the past, we showed differences in in vitro heterophil function between parental broilers (Line A>B and Line X>Y) and F1 reciprocal crosses (Line Cheterophil function. Specifically, the fast feathering gene (k) may enhance heterop...



Technology Transfer Automated Retrieval System (TEKTRAN)

In the past, we showed differences in heterophil function between parental broilers Lines A (fast [F] feathering)>B (slow [S] feathering) and their F1 reciprocal crosses (Line D [F]>C [S]), indicating the feathering gene is associated with heterophil function. In the present study, we evaluated the...



Technology Transfer Automated Retrieval System (TEKTRAN)

Recently, we showed differences in in vitro heterophil function between parental broilers (Line A>B) and F1 reciprocal crosses (Line Cheterophil functional efficiency while the slow feathering gene (K) reduced efficiency. Based on...


Neuronal synapse interaction reconstituted between live cells and supported lipid bilayers  

Microsoft Academic Search

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

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



Chemokine and cytokine levels in osteoarthritis and rheumatoid arthritis synovial fluid.  


To develop a method of the assay of chemokine and cytokine signaling in synovial fluid from patients suffering from osteoarthritis (OA) or rheumatoid arthritis (RA) and evaluate the effect of heterophilic antibodies on the reliability of the data. 21 synovial fluid samples from OA and 16 synovial fluid samples from RA patients were analyzed using a unique 2 step dot sandwich ELISA based micro-well protein array designed to detect heterophilic antibody signaling in the presence or absence of an effective heterophilic blocking reagent with assays carried out for Eotaxin, hGROa, interleukin (IL)-8, IP10, MCP-1, MCP-2, MIG, RANTES, TARC and IL-6. Array analysis reveals that the selective presence of heterophilic antibodies interferes with the accurate assay of synovial fluid samples from a minority of RA patients but not OA synovia. Using a commercial blocking diluent OA and RA synovial fluids reveal significant differences in chemokine content (IL-6, Eotaxin, hGROa, MCP-2, MIG, TARC, IL-8, RANTES). Using a two-step assay protocol it is possible to readily detect inappropriate antibody signaling due to heterophilic antibodies and devise a protocol designed to eliminate this problem thereby more accurately quantify cytokines and chemokines specific to both RA and OA fluids. PMID:23994256

Hampel, Ulrike; Sesselmann, Stefan; Iserovich, Pavel; Sel, Saadettin; Paulsen, Friedrich; Sack, Robert



Nectin ectodomain structures reveal a canonical adhesive interface  

PubMed Central

Nectins are immunoglobulin superfamily glycoproteins that mediate intercellular adhesion in many vertebrate tissues. Homophilic and heterophilic interactions between nectin family members help to mediate tissue patterning. We determined homophilic binding affinities and heterophilic specificities of all four nectins and the related protein nectin-like 5 from human and mouse, revealing a range of homophilic strengths and a defined heterophilic specificity pattern. To understand the molecular basis of adhesion and specificity, we determined crystal structures of natively glycosylated full ectodomains or adhesive fragments of nectins 1–4 and nectin-like 5. All crystal structures reveal dimeric nectins bound through a stereotyped interface previously proposed to represent a cis dimer. However, conservation of this interface and results of targeted cross-linking experiments show that this dimer likely represents the adhesive trans interaction. Its structure provides a simple molecular explanation for the adhesive binding specificity of nectins.

Harrison, Oliver J.; Vendome, Jeremie; Brasch, Julia; Jin, Xiangshu; Hong, Soonjin; Katsamba, Phinikoula S.; Ahlsen, Goran; Troyanovsky, Regina B.; Troyanovsky, Sergey M.; Honig, Barry; Shapiro, Lawrence



Nectin ectodomain structures reveal a canonical adhesive interface.  


Nectins are immunoglobulin superfamily glycoproteins that mediate intercellular adhesion in many vertebrate tissues. Homophilic and heterophilic interactions between nectin family members help mediate tissue patterning. We determined the homophilic binding affinities and heterophilic specificities of all four nectins and the related protein nectin-like 5 (Necl-5) from human and mouse, revealing a range of homophilic interaction strengths and a defined heterophilic specificity pattern. To understand the molecular basis of their adhesion and specificity, we determined the crystal structures of natively glycosylated full ectodomains or adhesive fragments of all four nectins and Necl-5. All of the crystal structures revealed dimeric nectins bound through a stereotyped interface that was previously proposed to represent a cis dimer. However, conservation of this interface and the results of targeted cross-linking experiments showed that this dimer probably represents the adhesive trans interaction. The structure of the dimer provides a simple molecular explanation for the adhesive binding specificity of nectins. PMID:22902367

Harrison, Oliver J; Vendome, Jeremie; Brasch, Julia; Jin, Xiangshu; Hong, Soonjin; Katsamba, Phinikoula S; Ahlsen, Goran; Troyanovsky, Regina B; Troyanovsky, Sergey M; Honig, Barry; Shapiro, Lawrence




PubMed Central

Epstein-Barr virus (EBV)-transformed autologous lymphoblasts were repeatedly inoculated into three squirrel monkeys. Each animal developed the heterophile antibodies of infectious mononucleosis and EBV-specific antibodies. After serologic responses had disappeared or markedly declined, the animals were challenged with either whole cells, cell filtrate, or cell ghosts. Animals challenged with living cells and cell ghosts developed agglutinin responses; the recipient of filtrate did not. The results suggest that EBV induces the appearance of the infectious mononucleosis heterophile antigen on the transformed cell membrane.

Shope, Thomas; Miller, George



Studies on an ascitic syndrome in young broilers 1. haematology and pathology  

Microsoft Academic Search

In the present investigation, the haematology and pathology of young broiler chicks with an ascitic syndrome were examined.Gross changes included cardiomegaly, ascites, congested lungs, shrunken livers, enlarged kidneys, dark breast muscle and congested intestines. Many blood parameters were significantly raised including the packed cell volume (PCV), haemoglobin (Hb), red (Rbc) and white blood cell (Wbc) counts. The heterophils and monocytes

M. H. Maxwell; G. W. Robertson; S. Spence




Technology Transfer Automated Retrieval System (TEKTRAN)

Nitric oxide is a rapidly reacting free radical which has cytotoxic effects during inflammatory responses and regulatory effects as a component of signal transduction cascades. We quantified the production of nitrite, a stable metabolite of nitric oxide, in chicken heterophils, monocytes and macrop...


Differential AP-1 and NF-kB expression in polymorphonuclear cells from a non-mammalian species following TLR4-mediated stimulation  

Technology Transfer Automated Retrieval System (TEKTRAN)

Previously, evaluation of heterophil function, susceptibility to pathogenic challenge, and pro-inflammatory cytokine mRNA expression levels showed differences between two parental lines of chickens. Line A is always more responsive and more resistant compared to Line B. Based on these findings, we...


Conjunctival xanthoma in a blue and gold macaw (Ara ararauna)  

Microsoft Academic Search

A 17-year-old female blue and gold macaw (Ara ararauna) presented for evaluation of a discreet, conjunctival mass of the OD. No other abnormalities were found on ophthalmic or physical examination. A heterophilic leukocytosis was present on the complete blood count, and elevated aspartate aminotransferase activity, creatinine kinase activity and cholesterol were present on the plasma biochemistry panel. Surgical removal of

Marcy J. Souza; Nancy S. Johnstone-McLean; Daniel Ward; Kimberly Newkirk



The neutrophil-specific antigen CD177 is a counter-receptor for platelet endothelial cell adhesion molecule-1 (CD31).  


Human neutrophil-specific CD177 (NB1 and PRV-1) has been reported to be up-regulated in a number of inflammatory settings, including bacterial infection and granulocyte-colony-stimulating factor application. Little is known about its function. By flow cytometry and immunoprecipitation studies, we identified platelet endothelial cell adhesion molecule-1 (PECAM-1) as a binding partner of CD177. Real-time protein-protein analysis using surface plasmon resonance confirmed a cation-dependent, specific interaction between CD177 and the heterophilic domains of PECAM-1. Monoclonal antibodies against CD177 and against PECAM-1 domain 6 inhibited adhesion of U937 cells stably expressing CD177 to immobilized PECAM-1. Transendothelial migration of human neutrophils was also inhibited by these antibodies. Our findings provide direct evidence that neutrophil-specific CD177 is a heterophilic binding partner of PECAM-1. This interaction may constitute a new pathway that participates in neutrophil transmigration. PMID:17580308

Sachs, Ulrich J H; Andrei-Selmer, Cornelia L; Maniar, Amudhan; Weiss, Timo; Paddock, Cathy; Orlova, Valeria V; Choi, Eun Young; Newman, Peter J; Preissner, Klaus T; Chavakis, Triantafyllos; Santoso, Sentot



Intercellular Adhesion Molecule-5 Induces Dendritic Outgrowth by Homophilic Adhesion  

PubMed Central

Intercellular adhesion molecule-5 (ICAM-5) is a dendritically polarized membrane glycoprotein in telencephalic neurons, which shows heterophilic binding to leukocyte ?2-integrins. Here, we show that the human ICAM-5 protein interacts in a homophilic manner through the binding of the immunoglobulin domain 1 to domains 4–5. Surface coated ICAM-5-Fc promoted dendritic outgrowth and arborization of ICAM- 5–expressing hippocampal neurons. During dendritogenesis in developing rat brain, ICAM-5 was in monomer form, whereas in mature neurons it migrated as a high molecular weight complex. The findings indicate that its homophilic binding activity was regulated by nonmonomer/monomer transition. Thus, ICAM-5 displays two types of adhesion activity, homophilic binding between neurons and heterophilic binding between neurons and leukocytes.

Tian, Li; Nyman, Henrietta; Kilgannon, Patrick; Yoshihara, Yoshihiro; Mori, Kensaku; Andersson, Leif C.; Kaukinen, Sami; Rauvala, Heikki; Gallatin, W. Michael; Gahmberg, Carl G.



Morphologic and cytochemical characteristics of blood cells from the desert tortoise (Gopherus agassizii).  


Morphologic and cytochemical staining characteristics of erythrocytes, leukocytes, and thrombocytes of the desert tortoise (Gopherus agassizii) were evaluated, using blood smears prepared from 23 healthy tortoises of Kern County, Calif. Special emphasis was placed on differentiating features of the various leukocytes and thrombocytes. A variety of cytochemical stains, including benzidine peroxidase, Sudan black B, chloroacetate esterase, alpha-naphthyl butyrate esterase, acid phosphatase, leukocyte alkaline phosphatase, periodic acid-Schiff, and toluidine blue were used. Heterophils had a characteristic, large, focal area of positive staining with chloroacetate esterase, alpha-naphthyl butyrate esterase, and acid phosphatase. Eosinophils stained diffusely positive with benzidine peroxidase, allowing differentiation of this leukocyte from heterophils. Thrombocytes stained focally positive with periodic acid-Schiff, allowing differentiation of these cells from lymphocytes, which stained uniformly negative. An intracytoplasmic body, commonly observed within erythrocytes, was considered ultrastructurally to represent a degenerate organelle. PMID:1384405

Alleman, A R; Jacobson, E R; Raskin, R E



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



Intracellular Campylobacter-like organisms associated with rectal prolapse and proliferative enteroproctitis in emus (Dromaius novaehollandiae).  


Rectal prolapse was the presenting clinical finding in a group of juvenile emus (Dromaius novaehollandiae). Gross findings included severely thickened and rugose distal rectal mucosae. Histologically, there were thickened villi, enterocyte hyperplasia, dilated glands filled with mucus and heterophils, and a dense infiltrate of heterophils, macrophages, lymphocytes, and plasma cells in the lamina propria. Examination of Warthin-Starry silver-stained sections revealed numerous apically located comma-shaped intracytoplasmic bacteria approximately 1 x 3 microns in size. Campylobacter-like organisms morphologically compatible with ileal symbiont intracellularis now known as Lawsonia intracellularis were seen via electron microscopy. Bacteria were further characterized by indirect immunofluorescence using monoclonal antibody specific for the 25-27-kd outer membrane protein of L. intracellularis. PMID:9066083

Lemarchand, T X; Tully, T N; Shane, S M; Duncan, D E



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



Leucocyte counts variation and measures of male fitness in the lekking Black Grouse  

Microsoft Academic Search

Differences in the ability to sustain chronic stress can underlie the variation in individuals’ fitness directly or indirectly\\u000a by impairing the expression of condition-dependent signals used in female mate choice. Therefore, understanding the effect\\u000a of the stress response on individuals’ fitness is key to identifying the mechanism of sexual selection. We tested the hypothesis\\u000a that an indicator of stress—the heterophil\\/lymphocyte

C. LebigreR; R. V. Alatalo; J. Kilpimaa; V. Staszewski; H. Siitari


Hematological stress indices reveal no effect of radio-transmitters on wintering Hermit Thrushes  

Microsoft Academic Search

Radio-telemetry is often used to track birds, and several investigators have examined the possible effects of radio-transmitters on birds. One approach to this question is to determine if transmitters induce physiological stress. Using hematological indicators of stress (heterophil-lymphocyte (H\\/L) ratios), studies of captive birds have revealed no evidence that radio-transmitters cause stress. However, studies in captivity may not reflect conditions

Andrew K. Davis; Nora E. Diggs; Robert J. Cooper; Peter P. Marra



Molecular Cloning and Characterization of Arginine-Specific ADP-Ribosyltransferases from Chicken Bone Marrow Cells  

Microsoft Academic Search

\\u000a Among a number of tissues and peripheral blood cells in chicken, leukocytes, bone marrow cells, liver and spleen showed high\\u000a ADP-ribosyltransferase activity, with leukocytes having the highest. Density gradient centrifugation of the leukocytes revealed\\u000a that the leukocyte ADP-ribosyltransferase originates in the polymorphonuclear cells, so called heterophils. Subcellular distribution\\u000a of the cells showed the localization of the enzyme in the granule

Makoto Shimoyama; Mikako Tsuchiya; Nobumasa Hara; Kazuo Yamada; Harumi Osago


Epstein-Barr Virus  

Microsoft Academic Search

Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus. Infection with EBV is common, worldwide in distribution, and largely\\u000a subclinical in early childhood. EBV has been established as the causative agent of heterophile-positive mononucleosis, which\\u000a occurs most frequently in late adolescence or early adulthood. In addition, seroepidemologic data have suggested that EBV\\u000a also plays an etiological role in African Burkitt’s lymphoma

Suresh B. Boppana


Determination of Cell Adhesion Sites of Neuropilin-1  

Microsoft Academic Search

Neuropilin-1 is a type 1 membrane protein with three distinct functions. First, it can mediate cell adhesion via a heterophilic molecular interaction. Sec- ond, in neuronal cells, neuropilin-1 binds the class 3 semaphorins, which are neuronal chemorepellents, and plays a role in the directional guidance of axons. Neuro- pilin-1 is expected to form complexes with the plexinA subfamily members and

Masayuki Shimizu; Yasunori Murakami; Fumikazu Suto; Hajime Fujisawa



Cytochemical staining and ultrastructural characteristics of peripheral blood leucocytes from the yellow rat snake ( Elaphe obsoleta quadrivitatta )  

Microsoft Academic Search

Cytochemical staining and ultrastructural characteristics of peripheral blood leucocytes from the yellow rat snake are described. A panel of cytochemical stains, including demonstration of myeloperoxidase, acid phosphatase, naphthol AS-D chloracetate esterase, alpha-naphthol butyrate esterase and alkaline phosphatase activities; and periodic acid-Schiff and Sudan Black-B staining was performed. Snake heterophils lacked peroxidase, alkaline phosphatase and acid phosphatase activity. Azurophils stained positively

D. I. Bounous; T. K. Dotson; R. L. Brooks Jr; E. C. Ramsay



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.



Structure and Interactions of NCAM Ig1-2-3 Suggest a Novel Zipper Mechanism for Homophilic Adhesion  

Microsoft Academic Search

The neural cell adhesion molecule, NCAM, mediates Ca2+-independent cell-cell and cell-substratum adhesion via homophilic (NCAM-NCAM) and heterophilic (NCAM-non-NCAM molecules) binding. NCAM plays a key role in neural development, regeneration, and synaptic plasticity, including learning and memory consolidation. The crystal structure of a fragment comprising the three N-terminal Ig modules of rat NCAM has been determined to 2.0 Å resolution. Based

Vladislav Soroka; Kateryna Kolkova; Jette S. Kastrup; Kay Diederichs; Jason Breed; Vladislav V. Kiselyov; Flemming M. Poulsen; Ingrid K. Larsen; Wolfram Welte; Vladimir Berezin; Elisabeth Bock; Christina Kasper



Amalgam, an axon guidance Drosophila adhesion protein belonging to the immunoglobulin superfamily: Over-expression, purification and biophysical characterization  

Microsoft Academic Search

Amalgam, a multi-domain member of the immunoglobulin superfamily, possesses homophilic and heterophilic cell adhesion properties. It is required for axon guidance during Drosophila development in which it interacts with the extracellular domain of the transmembrane protein, neurotactin, to promote adhesion. Amalgam was heterologously expressed in Pichia pastoris, and the secreted protein product, bearing an NH2-terminal His6Tag, was purified from the

Tzviya Zeev-Ben-Mordehai; Aviv Paz; Yoav Peleg; Lilly Toker; Sharon G. Wolf; Edwin H. Rydberg; Joel L. Sussman; Israel Silman



The crystal structure of the ligand-binding module of human TAG1 suggests a new mode of homophilic interaction  

Microsoft Academic Search

Human TAG-1 is a neural cell adhesion molecule that is crucial for the development of the nervous system during embryogenesis. It consists of six immunoglobulin-like and four fibronectin III-like domains and is anchored to the membrane by glycosylphosphatidylinositol. Herein we present the crystal structure of the four N-terminal immunoglobulin-like domains of TAG-1 (TAG-1Ig1-4), known to be important in heterophilic and

Mario Mörtl; Peter Sonderegger; Kay Diederichs; Wolfram Welte



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



Haematological health assessment in a passerine with extremely high proportion of basophils in peripheral blood  

Microsoft Academic Search

Haematological methods are widely utilised among avian ecologists as a means for individual health assessment. However, the\\u000a technical simplicity of some of the tests may easily lead to oversimplification of the evaluation. Here, we show in the Scarlet\\u000a Rosefinch (Carpodacus erythrinus) that haematological parameters other than the widely used heterophil\\/lymphocyte (H\\/L) ratio may be important to investigate.\\u000a We give the

Michal Vinkler; Jan Schnitzer; Pavel Munclinger; Jan Votýpka; Tomáš Albrecht



IgIII (270–280)-fragment-like H 2N-DDSDEEN-COOH peptide modulates N-CAM expression via Ca 2+-dependent ERK signaling during “in vitro neurogenesis”  

Microsoft Academic Search

The two major isoforms (180kDa and 140kDa) of the neural cell adhesion molecule (N-CAM) are crucially involved in neurogenesis and brain repair via activation of the mitogen-activated protein kinase (MAPK) cascade. Modification by glycosylation, and homophilic and heterophilic interactions regulate the function of N-CAM, but little is known about the interplay of these processes. In the neuron-like PC12 cell line,

Maria A. Mariggiò; Caterina Morabito; Simone Guarnieri; Antonietta Gentile; Kateryna Kolkova; Giorgio Fanò



Evaluation of C-reactive protein as an inflammatory biomarker in rabbits for vaccine nonclinical safety studies.  


INTRODUCTION: Inflammatory reactions are one of the potential safety concerns that are evaluated in the framework of vaccine safety testing. In nonclinical studies, the assessment of the inflammation relies notably on the measurement of biomarkers. C-reactive protein (CRP) is an acute-phase plasma protein of hepatic origin that could be used for that purpose in toxicity studies with rabbits. METHODS: To evaluate the utility of CRP as an additional inflammatory biomarker in adjuvant or vaccine toxicity studies, rabbits were injected on Day 0 with saline, aluminium phosphate, aluminium hydroxide, Adjuvant System (AS)01, AS03, AS15, or diphtheria-tetanus-whole cell pertussis-hepatitis B vaccine (DTPw-HB). Body weights, haematology parameters, CRP and fibrinogen levels were measured daily up to Day 7. Macroscopic changes at the injection site were also evaluated up to Day 7. At Day 7, a histopathological examination of the injection site was performed. RESULTS: Like fibrinogen, CRP levels rapidly increased after the injection of Adjuvant Systems or DTPw-HB, peaking at Day 1, and returning to baseline in less than a week. The magnitude of the CRP increase was consistently higher than that of fibrinogen with a larger fold increase from background, providing a more sensitive evaluation. The number of circulating heterophils was also increased on Day 1 after the injection of Adjuvant Systems or DTPw-HB. The highest increases in CRP levels were observed after the injection of DTPw-HB or AS03, and were also associated with the persistence of mixed inflammatory cell infiltrates (including heterophils) at the injection sites on Day 7. No increases in CRP levels and in circulating heterophils were observed after injection of the aluminium salt adjuvants. DISCUSSION: Our study supports the use of CRP as an accurate biomarker of acute inflammation in rabbits for vaccine toxicity studies and highlights an association between increased CRP levels and the recruitment of heterophils. PMID:23624216

Destexhe, Eric; Prinsen, Menk K; van Schöll, Inge; Kuper, C Frieke; Garçon, Nathalie; Veenstra, Stéphane; Segal, Lawrence



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.



Interactions of the cell adhesion molecule nectin with transmembrane and peripheral membrane proteins for pleiotropic functions  

Microsoft Academic Search

.  Cell adhesion molecules (CAMs) have been implicated in the control of a wide variety of cellular processes, such as cell adhesion,\\u000a polarization, survival, movement, and proliferation. Nectins have emerged as immunoglobulin-like CAMs that participate in\\u000a calcium-independent cell-cell adhesion by homophilic and heterophilic trans-interactions with nectins and nectin-like molecules. Nectin-based cell-cell adhesion exerts its function independently or\\u000a in cooperation with other

Y. Rikitake; Y. Takai



Vitamin E deficiency and pansteatitis in juvenile boat-billed herons (Cochlearius cochlearius).  


Steatitis due to vitamin E deficiency occurred in three 10-wk-old boat-billed herons (Cochlearius cochlearius) despite daily placement of a powdered vitamin supplement on the fish that was subsequently washed off by the parents. Physical findings included emaciation, yellow-brown subcutaneous nodules, a firm distended coelom, stomatitis, and yellow-white, submucosal pharyngeal nodules. Clinical pathology revealed heterophilic leukocytosis, anemia, hypoproteinemia, and low plasma alpha (alpha)-tocopherol levels (1.94 microg/ml and 2.14 microg/ml). Two of the chicks died of severe, diffuse pansteatitis and respiratory aspergillosis. PMID:10484150

Pollock, C G; Sleeman, J M; Houle, C D; Ramsay, E C



Pathogenicity of Mycoplasma lipofaciens strain ML64, isolated from an egg of a Northern Goshawk (Accipiter gentilis), for chicken embryos.  


Some Mycoplasma species are well-known avian pathogens and are of importance in poultry breeder flocks due to their pathogenic potential for embryos. Mycoplasmas are regularly detected in birds of prey, and a strain of Mycoplasma lipofaciens that was isolated from an egg of a Northern Goshawk (Accipiter gentilis) was examined for its pathogenicity in specific pathogen free chicken embryos since birds of prey eggs were not available for this purpose. The strain was found to be pathogenic, causing a high mortality as well as dwarfing, curled toes and infiltrations of heterophils in the liver, kidney, intestine and chorioallantoic membrane. PMID:17479376

Lierz, M; Stark, R; Brokat, S; Hafez, H M



Structural Biology of NCAM  

Microsoft Academic Search

\\u000a The neural cell adhesion molecule (NCAM) mediates Ca2+-independent cell-cell and cell-substratum adhesion via homophilic (NCAM-NCAM) and heterophilic (NCAM-non-NCAM molecules)\\u000a binding. NCAM is highly expressed in the nervous system and plays a key role in neural development, regeneration and synaptic\\u000a plasticity, including learning and memory consolidation. The questions in the structural biology of NCAM are mainly focused\\u000a on the mechanism of

Vladislav Soroka; Christina Kasper; Flemming M. Poulsen


An unusually high troponin I result in association with Legionella infection.  


A 77-year-old man presented himself with shortness of breath that was initially felt to be due to an acute coronary event, largely due to a very elevated troponin I result and his medical history. He subsequently showed evidence to suggest a significant pneumonia. The most likely candidate organism responsible, from the history and test results, appeared to be Legionella. We present the case for a spuriously and extremely elevated troponin I result, being at least in part due the production of heterophil antibodies by Legionella. PMID:24127373

Gill, Abhijit; Dubrey, Simon William



Some factors of immunopathogenesis of infectious mononucleosis in children.  


Sixty-eight children with the symptoms of infectious mononucleosis were examined. In 25 of them anti-EBV antibodies were found and in 10 heterophilic antibodies. In serum of 16 children sero-positive to EBV A, M, G, D immunoglobulins were determined as well as T and B lymphocytes. A statistically significant increase in the number of B-IgA, B-IgG and B-IgM lymphocytes, augmentation of T lymphocyte number and polyclonal hypergammaglobulinemia were detected. PMID:6293402

Rudkowski, Z; Kacprzak-Bergman, I; Jankowski, A; Krawczyk, K



Hematologic and plasma biochemistry values for endangered red knots (Calidris canutus rufa) at wintering and migratory sites in Argentina.  


We obtained hematologic and plasma biochemistry values for adult, long-distance migrant Red Knots at their southernmost wintering site in Río Grande (Tierra del Fuego, Argentina) and at the first stopover site in San Antonio Oeste (Río Negro, Argentina). Lymphocytes (L) followed by heterophils (H) were the most abundant leukocytes. H/L ratio and glucose levels were significantly higher at Río Grande, possibly because of the stress of migration and molting. Packed cell volume results ranged widely, probably in response to increased oxygen demand for migration. Protein profiles and lipids were higher at the stopover site and attributable to birds storing reserves for subsequent flights. PMID:20688666

D'Amico, Verónica L; Bertellotti, Marcelo; Baker, Allan J; González, Patricia M



Clinical and acquired immunologic responses to West Nile virus infection of domestic chickens (Gallus gallus domesticus).  


Numerous bird species are highly susceptible to North American strains of West Nile virus (WNV), and although domestic chickens are relatively resistant to WNV-associated disease, this species currently represents the most practical avian model for immune responses to WNV infection. Knowledge of the immunomodulation of susceptibility to WNV in birds is important for understanding taxonomic differences in infection outcomes. While focusing on immunophenotyping of CD3(+), CD4(+), CD8(+), and CD45(+) lymphocyte subpopulations, we compared lymphocyte subpopulations, blood chemistries, cloacal temperatures, IgM and IgG antibody titers, and differential whole-blood cell counts of WNV-infected and uninfected hens. Total blood calcium and lymphocyte numbers were lower in WNV-infected chickens compared with uninfected chickens. The heterophil-to-lymphocyte ratio increased over time from 2 to 22 d postinoculation (DPI) in uninfected chickens and from 2 to 8 DPI in WNV-infected chickens, although levels declined from 8 to 22 DPI in the latter group. No significant differences were found in the remaining immunological and hematological variables of the WNV-infected and uninfected groups. Our results reaffirm that chickens are resistant to WNV infection, and demonstrated that the heterophil-to-lymphocyte ratio differed between groups, allowing for sorting of infection status. Similar patterns in immune responses over time in both infected and uninfected hens may be related to age (i.e., 10 wk) and associated immune development. PMID:21248329

Fair, J M; Nemeth, N M; Taylor-McCabe, K J; Shou, Y; Marrone, B L



ALCAM/CD166 adhesive function is regulated by the tetraspanin CD9.  


ALCAM/CD166 is a member of the immunoglobulin superfamily of cell adhesion molecules (Ig-CAMs) which mediates intercellular adhesion through either homophilic (ALCAM-ALCAM) or heterophilic (ALCAM-CD6) interactions. ALCAM-mediated adhesion is crucial in different physiological and pathological phenomena, with particular relevance in leukocyte extravasation, stabilization of the immunological synapse, T cell activation and proliferation and tumor growth and metastasis. Although the functional implications of ALCAM in these processes is well established, the mechanisms regulating its adhesive capacity remain obscure. Using confocal microscopy colocalization, and biochemical and functional analyses, we found that ALCAM directly associates with the tetraspanin CD9 on the leukocyte surface in protein complexes that also include the metalloproteinase ADAM17/TACE. The functional relevance of these interactions is evidenced by the CD9-induced upregulation of both homophilic and heterophilic ALCAM interactions, as reflected by increased ALCAM-mediated cell adhesion and T cell migration, activation and proliferation. The enhancement of ALCAM function induced by CD9 is mediated by a dual mechanism involving (1) augmented clustering of ALCAM molecules, and (2) upregulation of ALCAM surface expression due to inhibition of ADAM17 sheddase activity. PMID:23052204

Gilsanz, Alvaro; Sánchez-Martín, Lorena; Gutiérrez-López, María Dolores; Ovalle, Susana; Machado-Pineda, Yesenia; Reyes, Raquel; Swart, Guido W; Figdor, Carl G; Lafuente, Esther M; Cabañas, Carlos



Neuronal synapse interaction reconstituted between live cells and supported lipid bilayers.  


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 beta-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. PMID:16408058

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



Pulmonary lesions in experimental ophidian paramyxovirus pneumonia of Aruba Island rattlesnakes, Crotalus unicolor.  


Histologic and ultrastructural changes were observed in the respiratory portions of lung in five 29-40-month-old Aruba Island rattlesnakes, Crotalus unicolor, that were inoculated with an Aruba Island Rattlesnake virus (AIV) strain of ophidian paramyxovirus (OPMV) isolated from an Aruba Island rattlesnake. Lungs from one non-infected and three mock-infected Aruba Island rattlesnakes were examined also. From 4 to 22 days following intratracheal inoculation, progressive microscopic changes were seen in the lung. Initially, increased numbers of heterophils were observed in the interstitium followed by proliferation and vacuolation of epithelial cells lining faveoli. The changes appeared to progress from cranial to caudal portions of the respiratory lung following inoculation. Beginning at 4 days postinoculation, viral antigen was demonstrated in epithelial cells lining faveoli with an immunofluorescent technique using a rabbit anti-AIV polyclonal antibody. Electron microscopy revealed loss of type I cells, hyperplasia of type II cells, and interstitial infiltrates of heterophils and mononuclear cells. Viral nucleocapsid material was seen within the cytoplasm and mature virus was seen budding from cytoplasmic membranes of infected type I and type II cells from 8 to 19 days after infection. A virus consistent with AIV was isolated from lung tissues of infected rattlesnakes, thus fulfilling Koch's postulates. PMID:9381656

Jacobson, E R; Adams, H P; Geisbert, T W; Tucker, S J; Hall, B J; Homer, B L



Capillariid nematodes in Brazilian turkeys, Meleagris gallopavo (Galliformes, Phasianidae): pathology induced by Baruscapillaria obsignata and Eucoleus annulatus (Trichinelloidea, Capillariidae).  


The pathology induced in turkeys (Meleagris gallopavo) by two capillariid nematodes, Baruscapillaria obsignata and Eucoleus annulatus is described together with data on prevalences, mean infection and range of worm burdens. B. obsignata occurred with a prevalence of 72.5% in the 40 examined hosts in a range of 2-461 nematodes and a mean intensity of 68.6, whereas E. annulatus was present in 2.5% of the animals, with a total amount of five recovered parasites. Gross lesions were not observed in the parasitized birds. Lesions due to B. obsignata mainly consisted of the thickening of intestinal villi with a mild mixed inflammatory infiltrate with the presence of mononuclear cells and heterophils. The lesions induced by E. annulatus were represented by foci of inflammatory infiltrate with heterophils in the crop epithelium and esophagus of a single infected female. These are the first pathological findings related to the presence of capillariid worms in turkeys to be reported in Brazil so far. Capillaria anatis, although present, was not pathogenic to the investigated turkeys. PMID:18545854

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



NTB-A Receptor Crystal Structure: Insights into Homophilic Interactions in the Signaling Lymphocytic Activation Molecule Receptor Family  

SciTech Connect

The signaling lymphocytic activation molecule (SLAM) family includes homophilic and heterophilic receptors that regulate both innate and adaptive immunity. The ectodomains of most SLAM family members are composed of an N-terminal IgV domain and a C-terminal IgC2 domain. NK-T-B-antigen (NTB-A) is a homophilic receptor that stimulates cytotoxicity in natural killer (NK) cells, regulates bactericidal activities in neutrophils, and potentiates T helper 2 (Th2) responses. The 3.0 {angstrom} crystal structure of the complete NTB-A ectodomain revealed a rod-like monomer that self-associates to form a highly kinked dimer spanning an end-to-end distance of {approx}100 {angstrom}. The NTB-A homophilic and CD2-CD58 heterophilic dimers show overall structural similarities but differ in detailed organization and physicochemical properties of their respective interfaces. The NTB-A structure suggests a mechanism responsible for binding specificity within the SLAM family and imposes physical constraints relevant to the colocalization of SLAM-family proteins with other signaling molecules in the immunological synapse.

Cao,E.; Ramagopal, U.; Fedorov, A.; Fedorov, E.; Yan, Q.; Lary, J.; Cole, J.; Nathenson, S.; Almo, S.



Epizootic podoknemidokoptiasis in American robins  

USGS Publications Warehouse

Epizootics of scaly leg disease caused by infection with the submacroscopic mite Knemidokoptes jamaicensis (Acari: Knemidokoptidae) in migratory American robins (Turdus migratorius) from a residential area of Tulsa (Oklahoma, USA) are documented during the winters (December through February) of 1993-94 and 1994-95. Estimates of 60 to > 80% of the birds in several different flights arriving in the area had lesions consistent with knemidokoptic mange. Epizootic occurrence of K. jamaicensis also is confirmed incidentally in American robins from Georgia (USA) in 1995 and 1998 and in Florida (USA) in 1991. These are the first confirmed epizootics of scaly leg attributed to infections with mites specifically identified as K. jamaicensis in North America. Severity of observed lesions in American robins ranged from scaly hyperkeratosis of the feet and legs to extensive proliferative lesions with loss of digits or the entire foot in some birds. Histologically, there was severe diffuse hyperkeratosis of the epidermis which contained numerous mites and multifocal aggregates of degranulating to degenerating eosinophilic heterophils; there was mild to severe superficial dermatitis with aggregates of eosinophilic heterophils and some mononuclear cells. Based on limited data from affected captive birds in Florida, we questioned the efficacy of ivermectin as an effective acaricide for knemidokoptiasis and propose that conditions associated with captivity may exacerbate transmission of this mite among caged birds. While knemidokoptic mange apparently can result in substantial host morbidity and possibly mortality, the ultimate impact of these epizootics on American robin populations presently is unknown.

Pence, D. B.; Cole, R. A.; Brugger, K. E.; Fischer, J. R.



Flagellin from Recombinant Attenuated Salmonella enterica Serovar Typhimurium Reveals a Fundamental Role in Chicken Innate Immunity  

PubMed Central

Recombinant attenuated Salmonella vaccines have been extensively studied, with a focus on eliciting specific immune responses against foreign antigens. However, very little is known about the innate immune responses, particularly the role of flagellin, in the induction of innate immunity triggered by recombinant attenuated Salmonella in chickens. In the present report, we describe two Salmonella enterica serovar Typhimurium vaccine strains, wild-type (WT) or flagellin-deficient (flhD) Salmonella, both expressing the fusion protein (F) gene of Newcastle disease virus. We examined the bacterial load and spatiotemporal kinetics of expression of inflammatory cytokine, chemokine, and Toll-like receptor 5 (TLR5) genes in the cecum, spleen, liver, and heterophils following oral immunization of chickens with the two Salmonella strains. The flhD mutant exhibited an enhanced ability to establish systemic infection compared to the WT. In contrast, the WT strain induced higher levels of interleukin-1? (IL-1?), CXCLi2, and TLR5 mRNAs in cecum, the spleen, and the heterophils than the flhD mutant at different times postinfection. Collectively, the present data reveal a fundamental role of flagellin in the innate immune responses induced by recombinant attenuated Salmonella vaccines in chickens that should be considered for the rational design of novel vaccines for poultry.

Pan, Zhiming; Cong, Qiuxia; Geng, Shizhong; Fang, Qiang; Kang, Xilong; You, Meng



[Persistent low hCG levels beyond pregnancy: report of two cases and review of the literature].  


Unexpected finding or persistence of low human chorionic gonadotropin (hCG) levels is not a rare situation. It requires a clinico-biological approach in order to avoid misunderstandings that could lead to inappropriate diagnostic or therapeutic attitudes. Beyond pregnancy, persistent low levels of hCG may be associated with various benign and malignant conditions, i.e. quiescent gestational trophoblastic disease (QTD), raised pituitary hCG or false positive elevation caused by circulating heterophile antibodies. We report the cases of two non-pregnant patients with low serum hCG. In the first case, hCG levels raised during several years following a spontaneous abortion. The likelihood of heterophilic antibodies interference was ruled out and extensive clinical investigation excluded the presence of a tumour. The diagnosis was QTD. In the second case, elevated hCG came to light as an incidental finding in a women with chronic renal failure and led the clinicians to question the laboratory. The cause was probably an increase in pituitary hCG consecutive to terminal renal failure. These cases illustrate the importance of understanding the biology of the hCG and the causes of its persistent low elevation, which are reviewed in this article. It is essential to demonstrate clinically the presence of a tumour in order to avoid unnecessary and ineffective chemotherapy and/or hysterectomy. PMID:23906583

De Backer, Benjamin; Goffin, Frédéric; Nisolle, Michelle; Minon, Jean-Marc


Functional analysis of posttranslational cleavage products of the neuron-glia cell adhesion molecule, Ng-CAM  

PubMed Central

Neuron-glia cell adhesion molecule (Ng-CAM) mediates cell adhesion between neurons homophilically and between neurons and glia heterophilically; it also promotes neurite outgrowth. In the chick brain, Ng-CAM is detected as glycoproteins of 190 and 210 kD (Ng- CAM200) with posttranslational cleavage products of 135 kD (F135, which contains most of the extracellular region) and 80 kD (F80, which includes the transmembrane and the cytoplasmic domains). To examine the functions of each of these components, we have expressed Ng-CAM200, F135, and F80 in murine L cells, and F135 and F80 as GST fusion proteins in the pGEX vector in bacteria. Appropriately transfected L cells expressed each of these proteins on their surfaces; F135 was also found in the media of cells transfected with Ng-CAM200 and F135. In addition to binding homophilically, cells transfected with Ng-CAM200 and F135 bound heterophilically to untransfected L cells, suggesting that there is a ligand for Ng-CAM on fibroblasts that may be related to the glial ligand. Detailed studies using the transfected cells and the fusion proteins indicated that both the homophilic and the heterophilic binding activities of Ng-CAM are localized in the F135 fragment of the molecule. The results also indicated that proteolytic cleavage of Ng- CAM200 is not required either for its expression on the cell surface or for cell adhesion and that there is an "anchor" for F135 on L cells (and presumably on neurons). In contrast to the cell binding results, the F80 but not the F135 fusion protein enhanced the outgrowth of neurites from dorsal root ganglion cells; this activity was associated with the FnIII repeats of F80. The observations that a protein corresponding to F135 contains the cell aggregation sites whereas one corresponding to the F80 has the ability to promote neurite outgrowth suggest that proteolytic cleavage may be an important event in regulating these Ng-CAM activities during embryonic development and neural regeneration.



Antinuclear antibodies in the sera of patients with nasopharyngeal carcinoma  

SciTech Connect

We studied the production of heterophile antinuclear antibodies (ANAs) in the sera of 50 patients, 20 with nasopharyngeal carcinoma (NPC) and 30 with other head and neck cancers (laryngeal cancer and maxillary cancer), before and after radiation therapy. A higher incidence of ANAs was found in the sera of patients with NPC and ANA production in these patients was higher after radiation therapy. We therefore performed in vitro experiments to explore the mechanisms of ANA production in the serum of postirradiated NPC patients. X-ray-irradiated NPC-derived cells (NPC-KT) produced a large amount of Epstein-Barr virus (NPC EBV) compared with non-irradiated NPC-KT cells. Nasopharyngeal carcinoma EBV-infected lymphocytes produced high levels of ANAs. These data suggest that lymphocytes infected by EBV from NPC cells may produce ANAs in the sera of NPC patients.

Takimoto, T.; Ishikawa, S.; Masuda, K.; Tanaka, S.; Yoshizaki, T.; Umeda, R. (Kanazawa Univ. (Japan))



Mycoplasma gallisepticum -induced release of macrophage inflammatory protein-1 beta from chicken monocytes-macrophages.  


Chicken monocytes and a macrophage-like cell line were used to determine the presence of macrophage inflammatory protein-1 beta (MIP-1 beta). RNA was extracted from these cells and subjected to reverse transcription with an anti-sense primer specific for the whole length of the MIP-1 beta cDNA. After a polymerase chain reaction to amplify the cDNA, a 200 bp gene product was detected, which corresponded to the molecular weight of the MIP-1 beta. The culture supernate of these cells did not have the ability to attract heterophils. However, after infection with Mycoplasma gallisepticum, a potent chemotactic substance in the culture supernate was detected. The chemotactic activity could be neutralized by antibodies prepared against two 10-amino-acid peptides of MIP-1 beta, indicating that the substance was MIP-1 beta. PMID:10627389

Lam, K M; DaMassa, A J



Molecular Mechanisms of Synaptic Specificity  

PubMed Central

Synapses are specialized junctions that mediate information flow between neurons and their targets. A striking feature of the nervous system is the specificity of its synaptic connections: an individual neuron will form synapses only with a small subset of available presynaptic and postsynaptic partners. Synaptic specificity has been classically thought to arise from homophilic or heterophilic interactions between adhesive molecules acting across the synaptic cleft. Over the past decade, many new mechanisms giving rise to synaptic specificity have been identified. Synapses can be specified by secreted molecules that promote or inhibit synaptogenesis, and their source can be a neighboring guidepost cell, not just presynaptic and postsynaptic neurons. Furthermore, lineage, fate, and timing of development can also play critical roles in shaping neural circuits. Future work utilizing large-scale screens will aim to elucidate the full scope of cellular mechanisms and molecular players that can give rise to synaptic specificity.

Margeta, Milica A.; Shen, Kang



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))



Effect of the naked neck gene on carcase composition and immunocompetence in chicken.  


1. An experiment was conducted to study the effect of naked neck gene (Na) on carcase measurements and some immunological responses of chickens. 2. The presence of naked neck gene in a single or double state increased body weight. 3. The naked neck genotypes (NaNa or Nana) exhibited higher relative weight of dressed carcase, drumstick and breast muscles compared to normally feathered individuals (nana). The proportion of abdominal fat was decreased in both naked neck genotypes compared with normally feathered ones. 4. A higher (non-specific) cell-mediated immune response was evoked in Na gene bearing birds. 5. Higher concentrations of antibodies occurred after injection of sheep red blood cells into naked neck birds, particularly at 14 d post-secondary injection. 6. The NaNa birds had a higher heterophil/lymphocyte ratio than Nana and nana siblings. PMID:18409083

Fathi, M M; El-Attar, A H; Ali, U M; Nazmi, A



Neurexin IV and Wrapper interactions mediate Drosophila midline glial migration and axonal ensheathment.  


Glia play crucial roles in ensheathing axons, a process that requires an intricate series of glia-neuron interactions. The membrane-anchored protein Wrapper is present in Drosophila midline glia and is required for ensheathment of commissural axons. By contrast, Neurexin IV is present on the membranes of neurons and commissural axons, and is highly concentrated at their interfaces with midline glia. Analysis of Neurexin IV and wrapper mutant embryos revealed identical defects in glial migration, ensheathment and glial subdivision of the commissures. Mutant and misexpression experiments indicated that Neurexin IV membrane localization is dependent on interactions with Wrapper. Cell culture aggregation assays and biochemical experiments demonstrated the ability of Neurexin IV to promote cell adhesion by binding to Wrapper. These results show that neuronal-expressed Neurexin IV and midline glial-expressed Wrapper act as heterophilic adhesion molecules that mediate multiple cellular events involved in glia-neuron interactions. PMID:19270173

Wheeler, Scott R; Banerjee, Swati; Blauth, Kevin; Rogers, Stephen L; Bhat, Manzoor A; Crews, Stephen T



Plasmodium circumflexum in a Shikra (Accipiter badius): phylogeny and ultra-structure of the haematozoa.  


A wild-caught, juvenile Shikra (Accipiter badius) was evaluated for rehabilitation at the Kasetsart University Raptor Rehabilitation Unit (KURRU) with a history of weakness. Plasmodium sp. was observed by both light and electron microscopy in blood obtained on day 1 of evaluation. Based on the appearance of erythrocytic meronts and gametocytes, the parasites were defined as Plasmodium (Giovannolaia) circumflexum. The sequence analysis of the mitochondrial cytochrome b gene from the plasmodia was closely related to parasites found in the Grey-headed woodpecker from Myanmar and the Brown hawk-owl from Singapore. Transmission electron microscopic examination revealed organelles in the haematozoa and heterophils that ingested the plasmodia. This is the first recorded case of Plasmodium circumflexum in a wild Shikra. This note emphasises the molecular characterisation and ultra-structure of the haematozoa. PMID:23094585

Salakij, Jarernsak; Lertwatcharasarakul, Preeda; Kasorndorkbua, Chaiyan; Salakij, Chaleow



Dermatitis and cellulitis in leopard geckos (Eublepharis macularius) caused by the Chrysosporium anamorph of Nannizziopsis vriesii.  


An epizootic of ulcerative to nodular ventral dermatitis was observed in a large breeding colony of 8-month to 5-year-old leopard geckos (Eublepharis macularius) of both sexes. Two representative mature male geckos were euthanized for diagnostic necropsy. The Chrysosporium anamorph of Nannizziopsis vriesii (CANV) was isolated from the skin lesions, and identification was confirmed by sequencing of the internal transcribed spacer region of the rRNA gene. Histopathology revealed multifocal to coalescing dermal and subcutaneous heterophilic granulomas that contained septate fungal hyphae. There was also multifocal epidermal hyperplasia with hyperkeratosis, and similar hyphae were present within the stratum corneum, occasionally with terminal chains of arthroconidia consistent with the CANV. In one case, there was focal extension of granulomatous inflammation into the underlying masseter muscle. This is the first report of dermatitis and cellulitis due to the CANV in leopard geckos. PMID:23160542

Toplon, D E; Terrell, S P; Sigler, L; Jacobson, E R



Cytochemical characterisation of the leucocytes and thrombocytes from Murray cod (Maccullochella peelii peelii, Mitchell).  


Cytochemistry has proven effective in differentiating specific cell lineages and elucidating their functional properties. This study utilised a range of cytochemical techniques to further investigate the leucocyte populations from Murray cod, an iconic Australian teleost fish species. This analysis provided clear insight into the structure and function of the leucocytes from this fish, which were found to be broadly similar to those of other fish species. However, some important differences were identified in Murray cod, such as the presence of naphthol AS chloroacetate esterase activity in the heterophil population, positive staining for periodic acid Schiff's, alkaline phosphatase and Sudan black B in the lymphocyte population, and a prevalent population of myeloid precursor cells. PMID:19332132

Shigdar, Sarah; Harford, Andrew; Ward, Alister C



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



Relating tumor score to hematology in green turtles with fibropapillomatosis in Hawaii  

USGS Publications Warehouse

The relationship between hematologic status and severity of tumor affliction in green turtles (Chelonia mydas) with fibropapillomatosis (FP) was examined. During 1 wk periods in July 1997 and July 1998, we bled 108 free-ranging green turtles from Pala'au (Molokai, Hawaii, USA) where FP is endemic. Blood was analyzed for hematocrit, estimated total solids, total white blood cell (WBC) count and differential WBC count. Each turtle was assigned a subjective tumor score ranging from 0 (no visible external tumors) to 3 (heavily tumored) that indicated the severity of FP. There was a progressive increase in monocytes and a decrease in all other hematologic parameters except heterophils and total numbers of white blood cells as tumor score increased. These data indicate that tumor score can relate to physiologic status of green turtles afflicted with FP, and that tumor score is a useful field monitor of severity of FP in this species.

Work, T. M.; Balazs, G. H.



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



The crystal structure of the ligand-binding module of human TAG-1 suggests a new mode of homophilic interaction  

PubMed Central

Human TAG-1 is a neural cell adhesion molecule that is crucial for the development of the nervous system during embryogenesis. It consists of six immunoglobulin-like and four fibronectin III-like domains and is anchored to the membrane by glycosylphosphatidylinositol. Herein we present the crystal structure of the four N-terminal immunoglobulin-like domains of TAG-1 (TAG-1Ig1–4), known to be important in heterophilic and homophilic macromolecular interactions. The contacts of neighboring molecules within the crystal were investigated. A comparison with the structure of the chicken ortholog resulted in an alternative mode for the molecular mechanism of homophilic TAG-1 interaction. This mode of TAG-1 homophilic interaction is based on dimer formation rather than formation of a molecular zipper as proposed for the chicken ortholog.

Mortl, Mario; Sonderegger, Peter; Diederichs, Kay; Welte, Wolfram



Potential sources of interference on Abeta immunoassays in biological samples  

PubMed Central

Therapeutic products that depend on the use of an in vitro diagnostic biomarker test to confirm their effectiveness are increasingly being developed. Use of biomarkers is particularly meaningful in the context of selecting the patient population where the therapeutic treatment is believed to be efficacious (patient enrichment). Currently available 'research-use-only' assays for Alzheimer's disease diagnosis all suffer from non-analyte and analyte-specific interferences. The impact of these interferences on the outcome of the assays is not well understood. The confounding factors are hampering correct value determination in biological samples and are intrinsic to the assay concept, the assay design, the presence in the sample of heterophilic antibodies and auto-antibodies, or might be the result of the therapeutic approach. This review focuses on the importance of assay interferences and considers how these might be minimized with the final aim of making the assays more acceptable as in vitro diagnostic biomarker tests for theranostic use.



Pathogenicity of Haemoproteus danilewskyi, Kruse, 1890, in blue jays (Cyanocitta cristata).  


Although the impact of blood parasite infections on passerine birds is potentially great, little is known of their pathologic effects. We studied Haemoproteus danilewskyi in experimentally infected captive and naturally infected free-ranging blue jays (Cyanocitta cristata) to determine patterns of infection and examine the pathologic effects of the parasite on the host. Physiologic changes, such as elevated numbers of lymphocytes, heterophils, basophils, eosinophils, and monocytes and decreased packed cell volume in the peripheral blood were associated with the erythrocytic phase of experimental infections of captive juvenile jays. Sublethal pathologic changes associated with the pre-erythrocytic phase of infections were observed in the liver, lung, and spleen. Schizonts were observed in the pulmonary capillaries of a 1 yr old jay necropsied 31 days post-inoculation, but not in 20 juvenile jays necropsied 57 days post-inoculation. In free-ranging naturally infected jays plasma protein concentration increased with density of natural infections. PMID:12685080

Garvin, Mary C; Homer, Bruce L; Greiner, Ellis C



Evidence-based approach for interpretation of Epstein-Barr virus serological patterns.  


Diagnosis of Epstein-Barr virus (EBV) infection is based on clinical symptoms and serological markers, including the following: immunoglobulin G (IgG) and IgM antibodies to the viral capsid antigen (VCA), heterophile antibodies, and IgG antibodies to the EBV early antigen-diffuse (EA-D) and nuclear antigen (EBNA-1). The use of all five markers results in 32 possible serological patterns. As a result, interpretation of EBV serologies remains a challenge. The purpose of this study was to use a large population of patients to develop evidence-based tools for interpreting EBV results. This study utilized 1,846 serum specimens sent to the laboratory for physician-ordered EBV testing. Chart review was performed for more than 800 patients, and diagnoses were assigned based on physician-ordered testing, clinical presentation, and patient history. Testing for all five EBV antibodies was performed separately on all serum samples using the Bio-Rad BioPlex 2200 system. Presumed EBV diagnosis (based on previous publications) was compared to EBV diagnosis based on a medical record review for each serological pattern. Interestingly, of the 32 possible serological patterns, only 12 occurred in > or = 10 patients. The remaining 20 patterns were uninterpretable because they occurred with such infrequency. Two easy-to-use tables were created to interpret EBV serological patterns based on whether three (EBV VCA IgG, IgM, and heterophile) or five markers are utilized. The use of these two tables allows for interpretation of >95% of BioPlex serological results. This is the first evidence-based study of its kind for EBV serology. PMID:19656988

Klutts, J S; Ford, B A; Perez, N R; Gronowski, A M



Evidence-Based Approach for Interpretation of Epstein-Barr Virus Serological Patterns ?  

PubMed Central

Diagnosis of Epstein-Barr virus (EBV) infection is based on clinical symptoms and serological markers, including the following: immunoglobulin G (IgG) and IgM antibodies to the viral capsid antigen (VCA), heterophile antibodies, and IgG antibodies to the EBV early antigen-diffuse (EA-D) and nuclear antigen (EBNA-1). The use of all five markers results in 32 possible serological patterns. As a result, interpretation of EBV serologies remains a challenge. The purpose of this study was to use a large population of patients to develop evidence-based tools for interpreting EBV results. This study utilized 1,846 serum specimens sent to the laboratory for physician-ordered EBV testing. Chart review was performed for more than 800 patients, and diagnoses were assigned based on physician-ordered testing, clinical presentation, and patient history. Testing for all five EBV antibodies was performed separately on all serum samples using the Bio-Rad BioPlex 2200 system. Presumed EBV diagnosis (based on previous publications) was compared to EBV diagnosis based on a medical record review for each serological pattern. Interestingly, of the 32 possible serological patterns, only 12 occurred in ?10 patients. The remaining 20 patterns were uninterpretable because they occurred with such infrequency. Two easy-to-use tables were created to interpret EBV serological patterns based on whether three (EBV VCA IgG, IgM, and heterophile) or five markers are utilized. The use of these two tables allows for interpretation of >95% of BioPlex serological results. This is the first evidence-based study of its kind for EBV serology.

Klutts, J. S.; Ford, B. A.; Perez, N. R.; Gronowski, A. M.



Evaluation of a multiplexed bead assay for assessment of Epstein-Barr virus immunologic status.  


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. PMID:15528686

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



Effects of genistein and hesperidin on biomarkers of heat stress in broilers under persistent summer stress.  


This study investigated the supplemental effects of the flavonoids genistein and hesperidin for biomarkers of heat stress in broilers reared under persistent summer stress. A total of 360 one-day-old, mixed-sex broiler chickens were divided into 6 treatment groups: control or supplemented with 5 mg of genistein•kg of feed(-1), 20 mg of hesperidin•kg of feed(-1), or a mixture of genistein and hesperidin (1:4) at a dosage of 5 mg•kg(-1), 10 mg•kg(-1), and 20 mg•kg(-1) of feed. Broilers were slaughtered at 42 d and samples were analyzed for hematological profile, creatine kinase, lactate dehydrogenase, and heat shock protein 70 mRNA levels. Results showed that dietary genistein and hesperidin improved (P < 0.05) the weekly performance of broilers particularly during the finisher period. The circulating heterophils and heterophil-to-lymphocyte ratios were found to decrease (P < 0.01) in the treated groups. Moreover, biomarkers of heat stress including the level of creatine kinase, lactate dehydrogenase, and heat shock protein 70 mRNA of breast muscle was also changed (P < 0.01) positively by the dietary compounds with pronounced effects of combined treatments. These findings suggested that genistein and hesperidin could be a prime strategy to ameliorate summer stress effects in broilers; and a combination of both compounds may lead to mutual synergistic effects. It could be suggested that dietary use of both genistein and hesperidin as a feed supplement may offer a potential nutritional strategy in tropical and subtropical regions to overcome the deleterious effects of persistent summer stress in broiler production. PMID:23960125

Kamboh, A A; Hang, S Q; Bakhetgul, M; Zhu, W-Y



Comparison of blood values and health status of Floreana Mockingbirds (Mimus trifasciatus) on the islands of Champion and Gardner-by-Floreana, Galápagos Islands.  


The Floreana Mockingbird (Mimus trifasciatus) is one of the rarest bird species in the world, with an estimated 550 individuals remaining on two rocky islets off the coast of Floreana, Galápagos, Ecuador, from which the main population was extirpated more than 100 yr ago. Because they have been listed in critical danger of extinction, a plan to reintroduce this species to Floreana has been initiated. Determining the health status of the source mockingbird populations is a top priority within the reintroduction plan. We report the health status, over the course of 4 yr, of 75 Floreana Mockingbirds on Champion Island and 160 Floreana Mockingbirds on Gardner-by-Floreana, based on physical examinations, hematology, hemolysis-hemagglutination assay, exposure to selected infectious disease agents, and ecto- and endoparasite counts. Birds on Gardner-by-Floreana had higher body condition index scores, packed cell volumes, total solids, and lymphocyte counts. Additionally, Gardner-by-Floreana birds had lower heterophil counts, eosinophil counts, and heterophil:lymphocyte ratios. No Chlamydophila psittaci DNA or antibodies to paramyxovirus-I, adenovirus-II, or Mycoplasma gallisepticum were found in any of the mockingbirds tested. Ectoparasites were present on birds from both islands, although species varied between islands. A coccidian species was found in eight of the 45 fecal samples from birds on Gardner-by-Floreana, but none of 33 birds examined from Champion. Birds on Gardner-by-Floreana were classified as healthier than those on Champion based on clinical and laboratory findings. These health data will be analyzed in conjunction with genetics, population structure, and disease presence on Floreana for developing recommendations for the Floreana Mockingbird reintroduction plan. PMID:21270000

Deem, Sharon L; Parker, Patricia G; Cruz, Marilyn B; Merkel, Jane; Hoeck, Paquita E A



Immunochemical studies of infectious mononucleosis. V. Isolation and characterization of a glycoprotein from goat erythrocyte membranes.  


A glycoprotein was isolated from goat erythrocyte membranes by extraction with hot 75% ethanol. The glycoprotein was purified by ethanol precipitation, phosphocellulose chromatography gel filtration, ethyl:ether and chloroform:methanol extraction. In aqueous phosphate-buffered saline, pH 7, the glycoprotein was in an aggregated state with a sedimentation coefficient (S(obs)) to 1.5. Electrophoresis of the glycoprotein on polyacrylamide gels containing phosphate-buffered 0.1% SDS gave a single band, staining with both periodic acid Schiff (PAS) and Coomassie Blue (CB). The apparent m.w., calculated from retardation coefficient, was 25,000. Electrophoresis of the glycoprotein on 1% SDS gels buffered with Tris-acetate (pH 7.4) showed a major band of similar (23,500) apparent m.w. plus four other PAS- and CB-staining bands of lower mobility. With 131I-labeled glycoprotein, recovery of bands from gels, sialic acid analysis, heterophile antigen activity, and re-electrophoresis, it was shown that these additional bands were aggregated forms of a single or closely related glycoprotein species. The purified glycoprotein contained 50% carbohydrate with molar ratios of sialic acid:galactose:mannose:galactosamine:glucosamine of 3.1:2.1:0.1:1.6:1. The glycoprotein was highly reactive with the Paul-Bunnell heterophile antibody in the sera of patients with infectious mononucleosis, with Limulus polyphemus lectin and weakly ractive with wheat germ agglutinin. These reactivities were destroyed by neuraminidase treatment or by alkaline sodium borohydride. The native glycoprotein did not react with lectins from Canavalia enisformis, Phaseolus vulgaris, Ricinus communis, or Vicia graminea although it was reactive with the latter two after neuraminidase treatment. PMID:956650

Fletcher, M A; Lo, T M; Graves, W R



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



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.



Effects of perch access and age on physiological measures of stress in caged White Leghorn pullets.  


The neuroendocrine system controls animals' adaptability to their environments by releasing psychotropic compounds such as catecholamines [epinephrine (EP), norepinephrine (NE), and dopamine (DA)], corticosterone (CORT), and serotonin (5-hydroxytryptamine or 5-HT). Changes of these neuroendocrine compounds have been used as biomarkers of animals' stress responses associated with their well-being. Assuming that pullets, like laying hens, are highly motivated to perch, we hypothesize that pullets with access to perches will experience less stress than pullets that never have access to perches. The objective of this study was to examine the effects of perch access and age on physiological measurements of stress in White Leghorn pullets housed in conventional cages. Hatchlings (n = 1,064) were randomly assigned to 28 cages. Two parallel metal round perches were installed in each of 14 cages assigned the perch treatment, whereas control cages were without perches. Two birds per cage were bled at wk 4, 6, and 12 wk of age. Plasma levels of CORT, DA, EP, and NE, blood concentrations of 5-HT and Trp, and heterophil to lymphocyte ratios were measured. Data were analyzed using a 2-way ANOVA. The perch treatment or its interaction with age did not affect any parameter measured in the study. The increase in the concentrations of circulating EP, NE, 5-HT (numerical increase at 4 wk), and Trp in 4- and 6-wk-old pullets compared with 12-wk-old pullets is unclear, but may have been due to acute handling stress at younger ages. In contrast, concentrations of DA were less at 4 wk compared with levels at 6 and 12 wk of age. Plasma CORT levels and the heterophil to lymphocyte ratio, indicators of long-term stress, were unaffected by age (P = 0.07 and 0.49, respectively). These results indicated that age, but not perch access, affects neuroendocrine homeostasis in White Leghorn pullets. Pullets that were never exposed to perches showed no evidence of eliciting a stress response. PMID:24135587

Yan, F F; Hester, P Y; Enneking, S A; Cheng, H W



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



The effect of feeder space allocation on productivity and physiology of Hy-Line W-36 hens housed in conventional cages.  


Insufficient feeder space for laying hens could increase competition at the feed trough, leading to disrupted feeding, inadequate nutrient intake, stress, and reduced productivity. The effects of feeder space allocation (FSA) on physiology and productivity were evaluated in beak-trimmed Hy-Line W-36 hens (n=480). They were obtained at 16.5 wk of age and housed on 4 tiers of shallow conventional cages. Five pullets/cage were housed at a stocking density of 434 cm2/hen and a feeder space of 12.2 cm/hen. After 1.5 wk of acclimation, baseline measurements were taken for feed utilization, bone mineralization, and heterophil:lymphocyte ratios. At 20 wk of age, pullets were given 5.8, 7.1, 8.4, 9.7, 10.9, or 12.2 cm of feeder space/bird (16 cages/treatment). Physiological and production measures were calculated monthly or twice a month for 12 mo. The heart, spleen, and right adrenal gland were collected from each hen at the end of the study. Data were analyzed using a repeated measures GLM incorporating cage, tier, FSA, and hen age. There were no effects of FSA on total egg production, bone mineral density, bone mineral content, heterophil:lymphocyte ratios, or organ weights. Hens with reduced FSA utilized more feed (P<0.001), had poorer feed conversion (P<0.001), and laid eggs with slightly thicker and heavier shells (P<0.001). There were effects of FSA on total egg weight (P<0.001) and hen-day egg production (P<0.001), but they were of low magnitude and not linear (P>0.05). Because BW was similar among FSA treatments, the results suggest that reduced feeder space did not limit feed intake. In addition, reduced FSA did not lower bone mineralization or cause physiological stress in W-36 hens housed in shallow cages, suggesting that it did not impair hen welfare. However, it did result in poorer feed efficiency, possibly related to greater feed wastage, predictive of an adverse economic effect from reducing feeder space. PMID:19687261

Thogerson, C M; Hester, P Y; Mench, J A; Newberry, R C; Okura, C M; Pajor, E A; Talaty, P N; Garner, J P



Effect of dietary prebiotic supplementation on the performance, intestinal microflora, and immune response of broilers.  


This study investigated the effects of dietary supplementation with the prebiotics fructo-oligosaccharide (FOS) and mannan-oligosaccharide (MOS) on the performance, small intestinal microflora, and immune response of broilers. Two hundred forty 1-d-old Ross broiler chickens were randomly assigned to 6 dietary treatment groups: control, avilamycin (6 mg/kg), 0.25% FOS, 0.5% FOS, 0.025% MOS, and 0.05% MOS. Each treatment was fed to 4 replicates of 10 birds per diet for 4 wk. Except for the 0.5% FOS group, the overall BW gains of birds treated with avilamycin and prebiotics were significantly(P < 0.05) higher than those of the control group. No significant differences were found between the control and supplemented groups in overall feed intake, feed conversion, and mortality. The 0.05% MOS group was significantly (P < 0.05) lower than the control and 0.5% FOS groups in heterophil:lymphocyte ratio and basophil level. Concentrations of plasma IgA and IgG were not significantly different among the treatment groups. Quantitative real-time PCR indicated that supplementation of the diet with avilamycin or prebiotics caused significant (P < 0.05) changes in the small intestinal microbial community, as determined in samples obtained at the ileocecal junction. The populations of Clostridium perfringens and Escherichia coli decreased with 0.25% FOS, 0.05% MOS, or avilamycin, and lactobacilli increased in the 0.25% FOS and 0.25% MOS treatment groups. Total bacteria increased in the 0.25% FOS and 0.05% MOS treatments and decreased in the avilamycin treatment. Feeding 0.25% FOS and 0.05% MOS resulted in an increase in lactobacillus community diversity in the ileum. Our results showed that 0.25% FOS and 0.05% MOS were comparable with avilamycin in improving productivity in broilers raised in wire floor cages up to 28 d of age. Plasma immunoglobulins were not affected by prebiotics, but the heterophil:lymphocyte ratio, basophil level, and microbial population in the ileum were significantly affected. PMID:21177446

Kim, G-B; Seo, Y M; Kim, C H; Paik, I K



Experimental production of ascites in broiler chickens using infectious bronchitis virus and Escherichia coli.  


Common commercial strain male broilers were intratracheally inoculated with 0.3 ml of fluid containing 10(3.7) embryo infective doses of infectious bronchitis virus (IBV) at 14 days of age and 7.5 x 10(6) colony-forming units of Escherichia coli at 18 days of age. Ascites was detected in 15 out of 100 infected birds, which was significantly higher than in a control group of 100 mock-infected birds (P < 0.01). Some parabronchi were blocked by copious exudate containing heterophils and fibrin in the infected birds at 22 days of age although these findings were not seen in the infected birds at 35 days of age or in birds with ascites. The erythrocyte packed cell volume and right ventricle/total ventricle (RV/TV) ratio of birds with ascites were higher than in birds without ascites. The RV/TV weight ratio for the infected group at the age of 35 days was higher than that of the control group. No IBV or E. coli were recovered from the ascitic birds. These findings suggest that these infectious agents induce ascites in the broilers, and then disappear until the birds suffer from ascites. PMID:9087339

Tottori, J; Yamaguchi, R; Murakawa, Y; Sato, M; Uchida, K; Tateyama, S


Differential adhesion and actomyosin cable collaborate to drive Echinoid-mediated cell sorting.  


Cell sorting involves the segregation of two cell populations into `immiscible' adjacent tissues with smooth borders. Echinoid (Ed), a nectin ortholog, is an adherens junction protein in Drosophila, and cells mutant for ed sort out from the surrounding wild-type cells. However, it remains unknown which factors trigger cell sorting. Here, we dissect the sequence of this process and find that cell sorting occurs when differential expression of Ed triggers the assembly of actomyosin cable. Conversely, Ed-mediated cell sorting can be rescued by recruitment of Ed, via homophilic or heterophilic interactions, to the wild-type cell side of the clonal interface, even when differential Ed expression persists. We found, unexpectedly, that when actomyosin cable was largely absent, differential adhesion was sufficient to cause limited cell segregation but with a jagged tissue border (imperfect sorting). We propose that Ed-mediated cell sorting is driven both by differential Ed adhesion that induces cell segregation with a jagged border and by actomyosin cable assembly at the interface that smoothens this border. PMID:21795280

Chang, Li-Hsun; Chen, Peilong; Lien, Mong-Ting; Ho, Yu-Huei; Lin, Chiao-Ming; Pan, Yi-Ting; Wei, Shu-Yi; Hsu, Jui-Chou



Amelioration of hepatotoxicity induced by aflatoxin using citrus fruit oil in broilers (Gallus domesticus).  


This study was conducted to evaluate the efficacy of citrus fruit oil (CFO; 2.5 g kg(-)(1)) on the clinicopathological changes in broilers fed with diets containing 1 ppm of aflatoxin (AF). A total of 160 Ross 308 broiler chicks of 1-day-old were procured from a commercial hatchery, divided randomly on 7th day of age into four groups with two replicates of 20 birds each and fed with basal diet (group A), basal diet + CFO (group B), basal diet + AF (group C) and CFO + basal diet + AF (group D). The gross and histopathological changes in the liver, kidney, spleen, thymus and bursa of Fabricius were investigated and relative organ weights were calculated. Slight to moderate hydropic degeneration, fatty change with the formation of cyst in some cases, periportal necrosis, infiltration of heterophils and mononuclear cells and bile duct hyperplasia were observed in chicks fed with 1 ppm AF-containing diet. The addition of CFO to AF-containing diet moderately decreased the magnitude and severity of lesions (hydropic degeneration and bile duct hyperplasia) in the liver. The supplementation of CFO to the basal diet did not produce any adverse effects in birds. PMID:23589408

Kumar, D Senthil; Rao, Suguna; Satyanaryana, Ml; Kumar, Pg Pradeep; Anitha, N



Dynamics of the systemic components of the chicken (Gallus gallus domesticus) immune system following activation by Escherichia coli; implications for the costs of immunity.  


The immune response is thought to be costly and deters from growth and reproduction, but the magnitude and sources of these costs are unknown. Thus, we quantified the changes in mass of leukocytes (CD4(+) and CD8(+) T cells, Bu1(+) IgM(+) and Bu1(+) IgG(+) B cells, monocytes/macrophages, heterophils and thrombocytes) and protective plasma proteins in systemic (non-mucosal) components of adult chickens injected intravenously with dead Escherichia coli. During the first day after E. coli injection most types of blood leukocytes decreased and ?-1-acid glycoprotein increased. Specific IgM, specific IgY, total IgM, Bu1(+) lymphocytes in the spleen and bone marrow and thymic CD8(+) lymphocytes increased at 5d post-injection. Quantitatively, the increases in the weight of cells and antibodies due to E. coli were dwarfed by the increase in the weight of the liver and acute phase proteins. Thus the acute phase response was markedly more costly than the subsequent adaptive response. The weight of the cells and proteins of the systemic immune system prior to challenge was 0.14% of body weight. Following E. coli injection, the additional weight of the immune components and the hypertrophy of the liver resulted in a 3.6-fold increase in weight which is equivalent to 18.5% of a large egg. PMID:23500513

Iseri, V J; Klasing, K C



State-dependent physiological maintenance in a long-lived ectotherm, the painted turtle (Chrysemys picta).  


Energy allocation among somatic maintenance, reproduction and growth varies not only among species, but among individuals according to states such as age, sex and season. Little research has been conducted on the somatic (physiological) maintenance of long-lived organisms, particularly ectotherms such as reptiles. In this study, we examined sex differences and age- and season-related variation in immune function and DNA repair efficiency in a long-lived reptile, the painted turtle (Chrysemys picta). Immune components tended to be depressed during hibernation, in winter, compared with autumn or spring. Increased heterophil count during hibernation provided the only support for winter immunoenhancement. In juvenile and adult turtles, we found little evidence for senescence in physiological maintenance, consistent with predictions for long-lived organisms. Among immune components, swelling in response to phytohemagglutinin (PHA) and control injection increased with age, whereas basophil count decreased with age. Hatchling turtles had reduced basophil counts and natural antibodies, indicative of an immature immune system, but demonstrated higher DNA repair efficiency than older turtles. Reproductively mature turtles had reduced lymphocytes compared with juvenile turtles in the spring, presumably driven by a trade-off between maintenance and reproduction. Sex had little influence on physiological maintenance. These results suggest that components of physiological maintenance are modulated differentially according to individual state and highlight the need for more research on the multiple components of physiological maintenance in animals of variable states. PMID:21147972

Schwanz, Lisa; Warner, Daniel A; McGaugh, Suzanne; Di Terlizzi, Roberta; Bronikowski, Anne



Rapid Expression of Chemokines and Proinflammatory Cytokines in Newly Hatched Chickens Infected with Salmonella enterica Serovar Typhimurium  

PubMed Central

Poultry meat and eggs contaminated with Salmonella enterica serovar Enteritidis or Salmonella enterica serovar Typhimurium are common sources of acute gastroenteritis in humans. However, the exact nature of the immune mechanisms protective against Salmonella infection in chickens has not been characterized at the molecular level. In the present study, bacterial colonization, development of pathological lesions, and proinflammatory cytokine and chemokine gene expression were investigated in the liver, spleen, jejunum, ileum, and cecal tonsils in newly hatched chickens 6, 12, 24, and 48 h after oral infection with Salmonella serovar Typhimurium. Very high bacterial counts were found in the ileum and cecal contents throughout the experiment, whereas Salmonella started to appear in the liver only from 24 h postinfection. Large numbers of heterophils, equivalent to neutrophils in mammals, and inflammatory edema could be seen in the lamina propria of the intestinal villi and in the liver. Interleukin 8 (IL-8), K60 (a CXC chemokine), macrophage inflammatory protein 1 ?, and IL-1? levels were significantly upregulated in the intestinal tissues and in the livers of the infected birds. However, the spleens of the infected birds show little or no change in the expression levels of these cytokines and chemokines. Increased expression of the proinflammatory cytokines and chemokines (up to several hundred-fold) correlated with the presence of inflammatory signs in those tissues. This is the first description of in vivo expression of chemokines and proinflammatory cytokines in response to oral infection with Salmonella in newly hatched chickens.

Withanage, G. S. K.; Kaiser, Pete; Wigley, Paul; Powers, Claire; Mastroeni, Pietro; Brooks, Heather; Barrow, Paul; Smith, Adrian; Maskell, Duncan; McConnell, Ian



Complementary Chimeric Isoforms Reveal Dscam1 Binding Specificity in Vivo  

PubMed Central

Summary Dscam1 potentially encodes 19,008 ectodomains of a cell recognition molecule of the immunoglobulin (Ig) superfamily through alternative splicing. Each ectodomain, comprising a unique combination of three variable (Ig) domains, exhibits isoform-specific homophilic binding in vitro. Although we have proposed that the ability of Dscam1 isoforms to distinguish between one another is crucial for neural circuit assembly, via a process called self-avoidance, whether recognition specificity is essential in vivo has not been addressed. Here we tackle this issue by assessing the function of Dscam1 isoforms with altered binding specificities. We generated pairs of chimeric isoforms that bind to each other (heterophilic) but not to themselves (homophilic). These isoforms failed to support self-avoidance or did so poorly. By contrast, co-expression of complementary isoforms within the same neuron restored self-avoidance. These data establish that recognition between Dscam1 isoforms on neurites of the same cell provides the molecular basis for self-avoidance.

Wu, Wei; Ahlsen, Goran; Baker, David; Shapiro, Lawrence; Zipursky, S. Lawrence



Comparison of Three Automated Immunoassay Methods for the Determination of Epstein-Barr Virus-Specific Immunoglobulin M ?  

PubMed Central

In this study we compared the performances of three commercially available Epstein-Barr virus (EBV) immunoglobulin M (IgM) assays on highly automated immunoassay platforms: BioPlex 2200 (Bio-Rad Laboratories), Immulite 2000 (Siemens Healthcare Diagnostics), and Liaison (DiaSorin). As a confirmatory method, immunoblotting was performed. The specificity of the three EBV IgM assays was evaluated by testing 293 selected sera from patients with various infectious and noninfectious diseases. After the exclusion of 30 samples, the specificities were 96.2% for Liaison, 98.1% for Immulite, and 97.0% for BioPlex. For evaluation of the sensitivity, samples from 70 consecutive patients with a positive heterophile antibody test were examined, irrespective of clinical or biological findings. After the exclusion of six samples, the sensitivities were 89.1% for Liaison, 84.4% for Immulite, and 89.1% for BioPlex. Finally, in a prospective study performed with 500 samples obtained from consecutive patients and sent in by general practitioners, we also determined Epstein-Barr nuclear antigen IgG and viral capsid antigen IgG in a two-phase approach. Concordance of the EBV serologic status was 96.2% between Liaison and Immulite, 96.4% between Immulite and BioPlex, and 97.8% between BioPlex and Liaison. The three EBV IgM immunoassays that we evaluated have acceptable and comparable performances.

Berth, Mario; Bosmans, Eugene



Amalgam, an axon guidance Drosophila adhesion protein belonging to the immunoglobulin superfamily: over-expression, purification and biophysical characterization.  


Amalgam, a multi-domain member of the immunoglobulin superfamily, possesses homophilic and heterophilic cell adhesion properties. It is required for axon guidance during Drosophila development in which it interacts with the extracellular domain of the transmembrane protein, neurotactin, to promote adhesion. Amalgam was heterologously expressed in Pichia pastoris, and the secreted protein product, bearing an NH(2)-terminal His(6)Tag, was purified from the growth medium by metal affinity chromatography. Size exclusion chromatography separated the purified protein into two fractions: a major, multimeric fraction and a minor, dimeric one. Two protocols to reduce the percentage of multimers were tested. In one, protein induction was performed in the presence of the zwitterionic detergent CHAPS, yielding primarily the dimeric form of amalgam. In a second protocol, agitation was gradually reduced during the course of the induction and antifoam was added daily to reduce the air/liquid interfacial foam area. This latter protocol lowered the percentage of multimer 2-fold, compared to constant agitation. Circular dichroism measurements showed that the dimeric fraction had a high beta-sheet content, as expected for a protein with an immunoglobulin fold. Dynamic light scattering and sedimentation velocity measurements showed that the multimeric fraction displays a monodisperse distribution, with R(H)=16 nm. When co-expressed together with amalgam the ectodomain of neurotactin copurified with it. Furthermore, both purified fractions of amalgam were shown to interact with Torpedo californica acetylcholinesterase, a structural homolog of neurotactin. PMID:18938249

Zeev-Ben-Mordehai, Tzviya; Paz, Aviv; Peleg, Yoav; Toker, Lilly; Wolf, Sharon G; Rydberg, Edwin H; Sussman, Joel L; Silman, Israel



Jamb and Jamc Are Essential for Vertebrate Myocyte Fusion  

PubMed Central

Cellular fusion is required in the development of several tissues, including skeletal muscle. In vertebrates, this process is poorly understood and lacks an in vivo-validated cell surface heterophilic receptor pair that is necessary for fusion. Identification of essential cell surface interactions between fusing cells is an important step in elucidating the molecular mechanism of cellular fusion. We show here that the zebrafish orthologues of JAM-B and JAM-C receptors are essential for fusion of myocyte precursors to form syncytial muscle fibres. Both jamb and jamc are dynamically co-expressed in developing muscles and encode receptors that physically interact. Heritable mutations in either gene prevent myocyte fusion in vivo, resulting in an overabundance of mononuclear, but otherwise overtly normal, functional fast-twitch muscle fibres. Transplantation experiments show that the Jamb and Jamc receptors must interact between neighbouring cells (in trans) for fusion to occur. We also show that jamc is ectopically expressed in prdm1a mutant slow muscle precursors, which inappropriately fuse with other myocytes, suggesting that control of myocyte fusion through regulation of jamc expression has important implications for the growth and patterning of muscles. Our discovery of a receptor-ligand pair critical for fusion in vivo has important implications for understanding the molecular mechanisms responsible for myocyte fusion and its regulation in vertebrate myogenesis.

Powell, Gareth T.; Wright, Gavin J.



Body condition and immune response in wild zebra finches: effects of capture, confinement and captive-rearing  

NASA Astrophysics Data System (ADS)

Behavioural ecologists attempt to predict fitness in birds from estimates of body condition and immune capacity. We investigated how the stresses associated with capture, confinement and captive-rearing of wild zebra finches (Taeniopygia guttata) affected different elements of the immune system and body condition. Wild birds had higher heterophil:lymphocyte ratios and total leucocyte counts than aviary birds, presumably an outcome of mounting specific resistance to pathogens, but this response diminished significantly within 10 days of confinement. Wild birds had lower phytohaemagglutinin-A (PHA) responses than their aviary-bred counterparts possibly because energetic costs limited a general resistance response. Wild birds were heavier and had higher haematocrits than their aviary counterparts, but had less fat, although just 10 days of captivity significantly increased fat levels. Measures of body condition were of limited use for predicting immune responsiveness. We conclude that the different elements of the immune system and body condition respond independently, and often unpredictably, to many ecological and behavioural stressors.

Ewenson, Erynne; Zann, Richard; Flannery, Graham



Inhibition of adrenal steroidogenesis, food restriction and acclimation to high ambient temperatures in chickens.  


1. White Plymouth Rock chickens placed under 60% food restriction or ad libitum feeding, with or without metyrapone treatment, from either 4 to 6 (early) or 24 to 26 (late) d of age were exposed to high ambient temperatures (35 +/- 2 degrees C) from 36 to 43 d of age. 2. Stress attributable to fasting was not manifested through leucocytic alteration when food-restricted chicks were supplemented with an adrenal blocking chemical, metyrapone. 3. Provision of metyrapone during the fasting period resulted in inferior compensatory growth during refeeding. 4. Exposure to high temperatures from 36 to 43 d of age did not cause an elevation in the heterophil:lymphocyte (H/L) ratios of chicks that had eaten metyrapone-treated food ad libitum during the neonatal stage. 5. During heat exposure, chicks that had been subjected to early 60% restriction with non-metyrapone-treated food had lower H/L ratios and improved resistance to marble spleen disease infection. PMID:7953785

Zulkifli, I; Dunnington, E A; Gross, W B; Siegel, P B



Carotenoid-based plumage coloration of male greenfinches reflects health and immunocompetence.  


Hypotheses of parasite-mediated sexual selection (PMSS) propose that elaborate male ornaments have evolved due to female preferences. Females would benefit from mating with more ornamental males if males' ornamentation signals their health status and ability to provide parasite resistance genes for the offspring. Carotenoid-based plumage coloration of birds has been hypothesised to honestly reflect an individual's health status due to trade-off in allocation of carotenoids between maintenance and signalling functions. The prediction of this hypothesis, namely that individuals with brighter plumage are able to mount stronger immune responses against novel antigens and reveal generally better health state, was tested in captive male greenfinches (Carduelis chloris). Greenfinches with brighter yellow breast feathers showed stronger humoral immune response against novel antigen (SRBC) while no relationship between plumage coloration and an estimate of cell-mediated immune responsiveness (PHA response) was detected. Elaborately ornamental individuals had better general health state as indicated by the negative correlations between plumage brightness and heterophil haemoconcentration. Consistent with the concept of PMSS, these results suggest that carotenoid-based plumage coloration in greenfinches honestly signals immunocompetence and health status. PMID:12647136

Saks, Lauri; Ots, Indrek; Hõrak, Peeter



Overexpression of the cell adhesion protein neuroligin-1 induces learning deficits and impairs synaptic plasticity by altering the ratio of excitation to inhibition in the hippocampus.  


Trans-synaptic cell-adhesion molecules have been implicated in regulating CNS synaptogenesis. Among these, the Neuroligin (NL) family (NLs 1-4) of postsynaptic adhesion proteins has been shown to promote the development and specification of excitatory versus inhibitory synapses. NLs form a heterophilic complex with the presynaptic transmembrane protein Neurexin (NRX). A differential association of NLs with postsynaptic scaffolding proteins and NRX isoforms has been suggested to regulate the ratio of excitatory to inhibitory synapses (E/I ratio). Using transgenic mice, we have tested this hypothesis by overexpressing NL1 in vivo to determine whether the relative levels of these cell adhesion molecules may influence synapse maturation, long-term potentiation (LTP), and/or learning. We found that NL1-overexpressing mice show significant deficits in memory acquisition, but not in memory retrieval. Golgi and electron microscopy analysis revealed changes in synapse morphology indicative of increased maturation of excitatory synapses. In parallel, electrophysiological examination indicated a shift in the synaptic activity toward increased excitation as well as impairment in LTP induction. Our results demonstrate that altered balance in the expression of molecules necessary for synapse specification and development (such as NL1) can lead to defects in memory formation and synaptic plasticity and outline the importance of rigidly controlled synaptic maturation processes. PMID:19437420

Dahlhaus, Regina; Hines, Rochelle M; Eadie, Brennan D; Kannangara, Timal S; Hines, Dustin J; Brown, Craig E; Christie, Brian R; El-Husseini, Alaa



Neurexin-Neuroligin Trans-Synaptic Interaction Mediates Learning-Related Synaptic Remodeling and Long-Term Facilitation in Aplysia  

PubMed Central

SUMMARY Neurexin and neuroligin, which undergo heterophilic interactions with each other at the synapse, are mutated in some patients with autism spectrum disorder, a set of disorders characterized by deficits in social and emotional learning. We have explored the role of neurexin and neuroligin at sensory-to-motor neuron synapses of the gill-withdrawal reflex in Aplysia that undergoes sensitization, a simple form of learned fear. We find that depleting neurexin in the presynaptic sensory neuron or neuroligin in the postsynaptic motor neuron abolishes both long-term facilitation and the associated presynaptic growth induced by repeated pulses of serotonin. Moreover, introduction into the motor neuron of the R451C mutation of neuroligin-3 linked to autism spectrum disorder blocks both intermediate-term and long-term facilitation. Our results suggest that activity-dependent regulation of the neurexin-neuroligin interaction may govern trans-synaptic signaling required for the storage of long-term memory, including emotional memory that may be impaired in autism spectrum disorder.

Choi, Yun-Beom; Li, Hsiu-Ling; Kassabov, Stefan R.; Jin, Iksung; Puthanveettil, Sathyanarayanan V.; Karl, Kevin A.; Lu, Yang; Kim, Joung-Hun; Bailey, Craig H.; Kandel, Eric R.



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


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



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.



Targeted mutation of Ncam to produce a secreted molecule results in a dominant embryonic lethality.  

PubMed Central

The neural cell adhesion molecule (NCAM) is a membrane-associated member of the immunoglobulin superfamily capable of both homophilic and heterophilic binding. To investigate the significance of this binding, a gene targeting strategy in embryonic stem (ES) cells was used to replace the membrane-associated forms of NCAM with a soluble, secreted form of its extracellular domain. Although the heterozygous mutant ES cells were able to generate low coat color chimeric mice, only the wild-type allele was transmitted, suggesting the possibility of dominant lethality. Analysis of chimeric embryos with high level of ES cell contribution revealed severe growth retardation and morphological defects by E8.5-E9.5. The second allele was also targeted, and embryos derived almost entirely from the homozygous mutant ES cells exhibited the same lethal phenotype as observed with heterozygous chimeras. Together, these results indicate that dominant lethality associated with the secreted NCAM does not require the presence of membrane-associated NCAM. Furthermore, the data indicate that potent bioactive cues or signals can be generated by NCAM. Images Fig. 1 Fig. 2 Fig. 3

Rabinowitz, J E; Rutishauser, U; Magnuson, T



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



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



The role and function of cadherins in the mammary gland  

PubMed Central

Cadherins are transmembrane receptors that function through calcium-dependent homophilic and heterophilic interactions that provide cell-cell contact and communication in many different organ systems. In the mammary gland only a few of the cadherins that make up this large superfamily of proteins have been characterized. Frequently in metastatic breast cancer, the genes for cadherins are epigenetically silenced, mutated, or regulated differently. During epithelial-mesenchymal transition, cadherins that are expressed normally in the epithelial cells are down-regulated, while cadherins expressed in the mesenchyme are up-regulated. This process is known as cadherin switching, and its regulation can sometimes facilitate the increased motility, invasiveness and proliferation that occurs in metastatic cancer cells. Depending on the context, however, cell motility, invasiveness, proliferation and expression of mesenchymal markers can be independently modulated from cadherin expression, leading to partial epithelial-mesenchymal transitions and even mesenchymal-epithelial transitions (METs). This review will summarize the current understanding of cadherins found in the mammary gland and what is known about their mechanism of regulation in the mammary gland during normal physiological conditions and in breast cancer.



Production of superoxide ions by leukocytes of the American alligator (Alligator mississippiensis).  


This study was conducted to characterize the production of superoxide ions by leukocytes in whole blood of the American alligator (Alligator mississippiensis). We used WST-1, a tetrazolium salt which can be reduced to a water-soluble formazan compound with high molar absorptivity at 438 nm, to probe the production of superoxide by alligator leukocytes. Incubation of alligator whole blood with WST-1 resulted in a time- and concentration-dependent increase in absorbance of the plasma at 438 nm. The reduction of WST-1 was inhibited in a concentration-dependent manner by superoxide dismutase, an enzyme that catalyzes the reduction of superoxide to peroxide, confirming that the reduction of WST-1 was due to the presence of superoxide. Treatment of whole blood with nitrotetrazolium blue (NBT) resulted in the staining of heterophils and monocytes, enforcing the idea that that the production of superoxide is due to the presence of leukocytes, and not other blood cell components. It is interesting to note that the production of superoxide by the alligator leukocytes required no external stimulation while human leukocytes must be stimulated with an immunological challenge before producing superoxide. This is the first report of the production of superoxide as an innate immune mechanism in crocodilians. PMID:18935969

Merchant, Mark; Williams, Stetson; Hardy, Ross



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

PubMed Central

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.



Pathomorphological, immunohistochemical and bacteriological findings in budgerigars (Melopsittacus undulatus) naturally infected with S. Gallinarum.  


The present study describes the pathological and bacteriological findings and diagnosis by immunoperoxidase and immunofluorescence methods in budgerigars (Melopsittacus undulatus) naturally infected with Salmonella gallinarum obtained from three commercial budgerigar rearing farms. The course of the disease in young budgerigars was peracute or acute, whereas in adult budgerigars the disease was acute or chronic. Clinically, yellow-white diarrhoea was observed in the young budgerigars with the acute form. In the adult budgerigars with the acute and chronic forms, a decrease in feed and water consumption with loss in body condition together with greenish-yellow diarrhoea was generally noted. Peritonitis and pericarditis were the most common findings in young budgerigars at necropsy, while in adult budgerigars scattered grey-white necrotic foci were found in the livers. Histopathologically, the lesions in young budgerigars were characterized with fibrinonecrotic peritonitis and/or pericarditis and necrotic hepatitis. In adult budgerigars with acute infection, hepatic necrosis with focal heterophil infiltration was present; whilst lesions in the chronic cases were granulomatous in nature with the infiltration of macrophages, lymphocytes and histiocytes. For the detection of S. Gallinarum in formalin-fixed, paraffin-embedded tissues, the avidin-biotin peroxidase complex and immunofluorescence methods were used. Both methods showed bacteria to be localized in the liver, kidney, peritoneum, heart, spleen and intestines of both young and adult budgerigars. The results of the present study indicate that the avidin-biotin peroxidase complex method was more sensitive than the immunofluorescence method in the detection of the bacteria. PMID:22515538

Tunca, R; Toplu, N; K?rkan, S; Avci, H; Aydo?an, A; Epikmen, E T; Tekbiyik, S



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



Neuroligin trafficking deficiencies arising from mutations in the alpha/beta-hydrolase fold protein family.  


Despite great functional diversity, characterization of the alpha/beta-hydrolase fold proteins that encompass a superfamily of hydrolases, heterophilic adhesion proteins, and chaperone domains reveals a common structural motif. By incorporating the R451C mutation found in neuroligin (NLGN) and associated with autism and the thyroglobulin G2320R (G221R in NLGN) mutation responsible for congenital hypothyroidism into NLGN3, we show that mutations in the alpha/beta-hydrolase fold domain influence folding and biosynthetic processing of neuroligin3 as determined by in vitro susceptibility to proteases, glycosylation processing, turnover, and processing rates. We also show altered interactions of the mutant proteins with chaperones in the endoplasmic reticulum and arrest of transport along the secretory pathway with diversion to the proteasome. Time-controlled expression of a fluorescently tagged neuroligin in hippocampal neurons shows that these mutations compromise neuronal trafficking of the protein, with the R451C mutation reducing and the G221R mutation virtually abolishing the export of NLGN3 from the soma to the dendritic spines. Although the R451C mutation causes a local folding defect, the G221R mutation appears responsible for more global misfolding of the protein, reflecting their sequence positions in the structure of the protein. Our results suggest that disease-related mutations in the alpha/beta-hydrolase fold domain share common trafficking deficiencies yet lead to discrete congenital disorders of differing severity in the endocrine and nervous systems. PMID:20615874

De Jaco, Antonella; Lin, Michael Z; Dubi, Noga; Comoletti, Davide; Miller, Meghan T; Camp, Shelley; Ellisman, Mark; Butko, Margaret T; Tsien, Roger Y; Taylor, Palmer



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



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



Associations between Organochlorine Contaminant Concentrations and Clinical Health Parameters in Loggerhead Sea Turtles from North Carolina, USA  

PubMed Central

Widespread and persistent organochlorine (OC) contaminants, such as polychlorinated biphenyls (PCBs) and pesticides, are known to have broad-ranging toxicities in wildlife. In this study we investigated, for the first time, their possible health effects on loggerhead sea turtles (Caretta caretta). Nonlethal fat biopsies and blood samples were collected from live turtles for OC contaminant analysis, and concentrations were compared with clinical health assessment data, including hematology, plasma chemistry, and body condition. Concentrations of total PCBs (?PCBs), ?DDTs, ?chlordanes, dieldrin, and mirex were determined in 44 fat biopsies and 48 blood samples. Blood concentrations of ?chlordanes were negatively correlated with red blood cell counts, hemoglobin, and hematocrit, indicative of anemia. Positive correlations were observed between most classes of OC contaminants and white blood cell counts and between mirex and ?TCDD-like PCB concentrations and the heterophil:lymphocyte ratio, suggesting modulation of the immune system. All classes of OCs in the blood except dieldrin were correlated positively with aspartate aminotransferase (AST) activity, indicating possible hepatocellular damage. Mirex and ?TCDD-like PCB blood concentrations were negatively correlated with alkaline phosphatase (ALP) activity. Significant correlations to levels of certain OC contaminant classes also suggested possible alteration of protein (?blood urea nitrogen, ?albumin:globulin ratio), carbohydrate (?glucose), and ion (?sodium, ?magnesium) regulation. These correlations suggest that OC contaminants may be affecting the health of loggerhead sea turtles even though sea turtles accumulate lower concentrations of OCs compared with other wildlife.

Keller, Jennifer M.; Kucklick, John R.; Stamper, M. Andrew; Harms, Craig A.; McClellan-Green, Patricia D.



Pathologic and hematologic responses to surgically implanted transmitters in eastern massasauga rattlesnakes (Sistrurus catenatus catenatus).  


The study of secretive snakes, such as rattlesnakes, has benefited from the use of radiotelemetry. However, the principal assumption in telemetry studies is that the transmitter has no significant effect on the study animal. To test the validity of this assumption, the physiologic and pathologic effects of intracoelomic implants were examined in a group of 24 eastern massasauga rattlesnakes (Sistrurus catenatus catenatus) in a laboratory setting over a period of 58 wk between March 2005 and April 2006. Inflammation and infection were evaluated using gross examination, histopathology, bacteriology, hematology, and plasma protein electrophoresis. Inflammation and infection occurred despite careful surgical procedures and advanced veterinary care. Four of 12 (33%) snakes developed extensive inflammatory response to the transmitter and associated anaerobic and gram-negative bacterial infections. Another four (33%) snakes showed mild inflammatory responses without infection. Reaction to the transmitters was reflected in changes in values for heterophils, monocytes, alpha-1, and beta globulin levels. Some conclusions reached in field studies using implanted radiotransmitters in snakes may be invalid if the implant influences the behavior or survival of the subject. Advances in attachment methods and transmitter coating technology may prevent some of the adverse effects associated with surgically implanted transmitters. PMID:21270001

Lentini, Andrew M; Crawshaw, Graham J; Licht, Lawrence E; McLelland, David J



Scholarly tailgating defined: A diverse, giant network.  


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. The concept of scholarly tailgating explains the use of homophilic and diverse, multidisciplinary network relationships to foster an intellectual community in healthcare, and to benefit healthcare consumer. 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. PMID:23589967

Adegbola, Maxine



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

PubMed Central

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.

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



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



Chronic dermatitis caused by Lactobacillus jensenii infection in a blue and gold macaw (Ara ararauna).  


Case Description-A 5-year-old sexually intact female blue and gold macaw (Ara ararauna) was evaluated because of a swelling on the right side of the face and irritated area on the ventral aspect of the keel. Clinical Findings-Clinical findings were consistent with dermatitis (right facial lesion) and a coalescing subdermal granuloma (ventral keel lesion). Hematologic analysis revealed monocytosis and mild anemia. Histologic evaluation of the ventral keel lesion revealed evidence of chronic heterophilic dermatitis with multinucleated giant cells and bacterial rods and cocci. An unspeciated gram-positive rod-shaped bacterium was isolated via aerobic bacterial culture. Results of bacterial biochemical tests suggested the organism was a type of Actinomyces. A 16S rRNA gene sequence analysis was performed; results indicated the organism was Lactobacillus jensenii. Treatment and Outcome-Extensive surgical debridement of the branching granuloma, which extended throughout the length of the keel, followed by long-term treatment with ciprofloxacin and clindamycin provided full resolution of clinical signs. No recrudescence of clinical signs was evident for up to 18 months after the initial evaluation. Clinical Relevance-To the authors' knowledge, this is the first report of Lactobacillus-associated dermatitis or subdermal granuloma in the scientific literature and the second report of L jensenii in avian species. Use of 16S rRNA gene sequence analysis was instrumental in the identification of this fastidious organism, indicating the method's usefulness as a diagnostic tool. PMID:24050571

Brandão, João; Wong, Carolyn; Kurotaki, Tetsuro; Johnson, James; Mitchell, Maria; Roy, Alma; Pucheu-Haston, Cherie; Del Piero, Fabio; Tully, Thomas



Cooperative Plasminogen Recruitment to the Surface of Streptococcus canis via M Protein and Enolase Enhances Bacterial Survival  

PubMed Central

ABSTRACT Streptococcus canis is a zoonotic pathogen capable of causing serious invasive diseases in domestic animals and humans. Surface-exposed M proteins and metabolic enzymes have been characterized as major virulence determinants in various streptococcal species. Recently, we have identified SCM, the M-like protein of S. canis, as the major receptor for miniplasminogen localized on the bacterial surface. The present study now characterizes the glycolytic enzyme enolase as an additional surface-exposed plasminogen-binding protein. According to its zoonotic properties, purified S. canis enolase binds to both human and canine plasminogen and facilitates degradation of aggregated fibrin matrices after activation with host-derived urokinase-type plasminogen activator (uPA). Unlike SCM, which binds to the C terminus of human plasminogen, the S. canis enolase interacts N terminally with the first four kringle domains of plasminogen, representing angiostatin. Radioactive binding analyses confirmed cooperative plasminogen recruitment to both surface-exposed enolase and SCM. Furthermore, despite the lack of surface protease activity via SpeB in S. canis, SCM is released and reassociated homophilically to surface-anchored SCM and heterophilically to surface-bound plasminogen. In addition to plasminogen-mediated antiphagocytic activity, reassociation of SCM to the bacterial surface significantly enhanced bacterial survival in phagocytosis analyses using human neutrophils.

Fulde, Marcus; Rohde, Manfred; Polok, Andy; Preissner, Klaus T.; Chhatwal, Gursharan Singh; Bergmann, Simone



Expression of gicerin in development, oncogenesis and regeneration of the chick kidney.  


Neurite outgrowth factor, which promotes neurite extension from neuronal cells, is an extracellular matrix glycoprotein belonging to the laminin family. Gicerin is a protein that binds neurite outgrowth factor. Its cDNA cloning has revealed that it is a novel cell adhesion molecule belonging to the immunoglobulin super-family. Functional analysis demonstrates that gicerin possesses homophilic binding activity as well as heterophilic binding activity with neurite outgrowth factor. We examined the role and expression of neurite outgrowth factor and gicerin in chick kidney during development. In the embryonic kidney, gicerin was found to be highly expressed both on ureteric bud cells and metanephrogenic mesenchymal cells, when the mesenchymal cells become condensed to be converted into polarized epithelial cells. In the adult kidney, the expression of gicerin was decreased and restricted to the glomerulus, proximal tubule and medullary loop. On the other hand, neurite outgrowth factor was constitutively expressed in the basement membranes of tubules and the matrices of glomeruli during development. As some molecules which are expressed during embryogenesis and suppressed after maturation are re-expressed in tumor cells or tissues during regeneration, we also examined the expression of gicerin in chicken Wilms' tumor and regenerating kidney in interstitial nephritis. Gicerin was remarkably upregulated in Wilms' tumor and re-expressed in collecting ducts recovering from interstitial nephritis. These findings suggest that gicerin could play a role not only in normal renal development but also in oncogenesis and regeneration. PMID:7622008

Takaha, N; Taira, E; Taniura, H; Nagino, T; Tsukamoto, Y; Matsumoto, T; Kotani, T; Sakuma, S; Miki, N



Structural insights into the exquisite selectivity of neurexin/neuroligin synaptic interactions  

PubMed Central

The extracellular domains of neuroligins and neurexins interact through Ca2+ to form flexible trans-synaptic associations characterized by selectivity for neuroligin or neurexin subtypes. This heterophilic interaction, essential for synaptic maturation and differentiation, is regulated by gene selection, alternative mRNA splicing and post-translational modifications. A new, 2.6 Å-resolution crystal structure of a soluble neurexin-1?–neuroligin-4 (Nrx1?–NL4) complex permits a detailed description of the Ca2+-coordinated interface and unveils concerted positional rearrangements of several residues of NL4, not observed in neuroligin-1, associated with Nrx1? binding. Surface plasmon resonance analysis of the binding of structure-guided Nrx1? mutants towards NL4 and neuroligin-1 shows that flexibility of the Nrx1?-binding site in NL4 is reflected in a greater dissociation constant of the complex and higher sensitivity to ionic strength and pH variations. Analysis of neuroligin mutants points to critical functions for two respective residues in neuroligin-1 and neuroligin-2 in governing the affinity of the complexes. Although neuroligin-1 and neuroligin-2 have pre-determined conformations that respectively promote and prevent Nrx1? association, unique conformational reshaping of the NL4 surface is required to permit Nrx1? association.

Leone, Philippe; Comoletti, Davide; Ferracci, Geraldine; Conrod, Sandrine; Garcia, Simon U; Taylor, Palmer; Bourne, Yves; Marchot, Pascale



Effects of selenium on mallard duck reproduction and immune function  

SciTech Connect

Selenium from irrigation drain water and coal-fired power stations is a significant environmental contaminant in some regions of the USA. The objectives were to examine whether selenium-exposed waterfowl had altered immune function, disease resistance, or reproduction. Pairs of adult mallards were exposed for 95-99 days on streams with sodium selenite-treated water at 10 and 30 ppb, or on untreated streams. Selenium biomagnified through the food chain to the ducks. Disease resistance was decreased in ducklings hatched on the streams and challenged with duck hepatitis virus 1 (DHV1) when 15-days old. Liver selenium concentrations for these ducklings on the 10 and 30 ppb streams was 3.6 and 7.6 ppm dry weight, respectively. Mortality of ducklings purchased when 7-days old, exposed to selenium for 14 days, and challenged when 22-days old was not affected. However, their selenium exposure was lower (liver selenium 4.1 ppm dry weight for the 30 ppb stream). Five parameters of immune function were measured in adult ducks. Phagocytosis of killed Pasteurella multocida by blood heterophils and monocytes, and blood monocyte concentrations were higher in adult males following 84 days exposure to 30 ppb selenium. Their liver selenium concentrations were 11.1 ppm dry weight after 95-99 days exposure.

Whiteley, P.L.; Yuill, T.M.; Fairbrother, A.



Performance and physiological parameters of broiler chickens subjected to fasting on the neonatal period.  


Broiler chicks aged 12 h after hatching were allotted according to a block design in a 7 x 2 factorial schedule of 14 treatments and four replications of 50 chicks each one. The main experimental factors were fasting for 0, 6, 12, 18, 24, 30, and 36 h after chick placement and sex. Independent of sex, fasting had a negative linear effect on weight and productivity of broilers at market age (42 d) without affecting feed conversion or mortality index. Groups subjected to 18 and 36 h of fasting after placement, corresponding to 30 and 48 h posthatching fasting, had lower biometrical values for small intestine (length, weight, and size; villus height; and crypt depth) than chicks fed immediately after placement. According to the Pearson test, BW of birds at 21 and 42 d were significantly correlated to BW at 7 d (r=0.77) and 21 d (r=0.45), respectively. Males performed better than females but had higher mortality rates. Fasting did not influence serum concentrations of corticosterone or sexual steroid hormones. Nevertheless, early signs of sexual dimorphism arose from the high estradiol (E2) concentration on female serum. Heterophil:lymphocyte ratio was not different among treatments, indicating that early fasting did not seem to be a stress factor 21 or 42 d after fasting. The results suggested a maximum fasting of 24 h after hatching in order to preserve broiler productivity at market age. PMID:12943295

Gonzales, E; Kondo, N; Saldanha, E S P B; Loddy, M M; Careghi, C; Decuypere, E



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



Discospondylitis caused by Staphylococcus aureus in an African black-footed penguin (Spheniscus demersus).  


A 22-year-old female African black-footed penguin (Spheniscus demersus), housed indoors with other African and rockhopper penguins, was presented acutely with lethargy, ataxia, and hind limb weakness after a molt. The penguin would assume a hunched position and, when resting, sat on its hocks or lay on its keel. Physical and neurologic examination revealed hind limb paraparesis, proprioceptive deficits, and tiptoe walking. Results of a complete blood cell count and biochemical analysis revealed mild heterophilic leukocytosis, anemia, mild hypoalbuminemia, hypokalemia, and hyperuricemia. Results of whole-body radiographs and coelioscopy were unremarkable. Two computed tomographies of the spine at a 3-month interval revealed a lesion at the mobile thoracic vertebra proximal to the synsacrum with associated spinal cord compression. The penguin was treated with itraconazole, doxycycline, and meloxicam, and it initially improved with return to near normal gait and behavior. However, 5 months after the onset of clinical signs, the penguin was euthanatized after a relapse with worsening of the neurologic signs. Postmortem and histopathologic examination revealed focal granulomatous discospondylitis at the penultimate mobile thoracic vertebra, with intralesional bacteria from which Staphylococcus aureus was cultured. PMID:23409435

Field, Cara L; Beaufrère, Hugues; Wakamatsu, Nobuko; Rademacher, Nathalie; MacLean, Robert



Generation and Nuclear Translocation of Sumoylated Transmembrane Fragment of Cell Adhesion Molecule L1  

PubMed Central

The functions of the cell adhesion molecule L1 in the developing and adult nervous system are triggered by homophilic and heterophilic interactions that stimulate signal transductions that activate cellular responses. Here, we show that stimulation of signaling by function-triggering L1 antibodies or L1-Fc leads to serine protease-dependent cleavage of full-length L1 at the plasma membrane and generation of a sumoylated transmembrane 70-kDa fragment comprising the intracellular and transmembrane domains and part of the extracellular domain. The 70-kDa transmembrane fragment is transported from the plasma membrane to a late endosomal compartment, released from endosomal membranes into the cytoplasm, and transferred from there into the nucleus by a pathway that depends on importin and chromatin-modifying protein 1. Mutation of the sumoylation site at Lys1172 or of the nuclear localization signal at Lys1147 abolished L1-stimulated generation or nuclear import of the 70-kDa fragment, respectively. Nuclear import of the 70-kDa fragment may activate cellular responses in parallel or in association with phosphorylation-dependent signaling pathways. Alterations in the levels of the 70-kDa fragment during development and in the adult after spinal cord injury or in a mouse model of Alzheimer disease suggest that this fragment is functionally implicated in development, regeneration, neurodegeneration, tumorigenesis, and possibly synaptic plasticity in the mature nervous system.

Lutz, David; Wolters-Eisfeld, Gerrit; Joshi, Gunjan; Djogo, Nevena; Jakovcevski, Igor; Schachner, Melitta; Kleene, Ralf



A Genetically Encoded Tag for Correlated Light and Electron Microscopy of Intact Cells, Tissues, and Organisms  

PubMed Central

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.

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



The desmosome and pemphigus  

PubMed Central

Desmosomes are patch-like intercellular adhering junctions (“maculae adherentes”), which, in concert with the related adherens junctions, provide the mechanical strength to intercellular adhesion. Therefore, it is not surprising that desmosomes are abundant in tissues subjected to significant mechanical stress such as stratified epithelia and myocardium. Desmosomal adhesion is based on the Ca2+-dependent, homo- and heterophilic transinteraction of cadherin-type adhesion molecules. Desmosomal cadherins are anchored to the intermediate filament cytoskeleton by adaptor proteins of the armadillo and plakin families. Desmosomes are dynamic structures subjected to regulation and are therefore targets of signalling pathways, which control their molecular composition and adhesive properties. Moreover, evidence is emerging that desmosomal components themselves take part in outside-in signalling under physiologic and pathologic conditions. Disturbed desmosomal adhesion contributes to the pathogenesis of a number of diseases such as pemphigus, which is caused by autoantibodies against desmosomal cadherins. Beside pemphigus, desmosome-associated diseases are caused by other mechanisms such as genetic defects or bacterial toxins. Because most of these diseases affect the skin, desmosomes are interesting not only for cell biologists who are inspired by their complex structure and molecular composition, but also for clinical physicians who are confronted with patients suffering from severe blistering skin diseases such as pemphigus. To develop disease-specific therapeutic approaches, more insights into the molecular composition and regulation of desmosomes are required.



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



Humoral and cell-mediated immune function in adult Japanese Quail following exposure to 2. 45-GHz microwave radiation during embryogeny  

SciTech Connect

Japanese quail, Coturnix coturnix japonica, eggs were subjected to 2.45-GHz CW microwave radiation at 5 mW/cm2 (SAR . 4.03 mW/g) during the first 12 days of embryogeny. Following hatching the exposed embryos, as well as nonexposed controls, were reared to 22 weeks of age. Humoral immune potential, as indicated by comparable anti-CRBC antibody, IgM and IgG, levels at 0, 4, and 7 days postimmunization in both exposed and control quail was not affected significantly. However, cell-mediated immune potential, measured by the reaction to intradermal injection of phytohemagglutinin-P in the wing web, was reduced in the exposed females, but not in the exposed males. Additionally, total leukocyte numbers and absolute circulating numbers of lymphocytes, monocytes, and heterophils were increased significantly only in the exposed females. These data show that exposure of Japanese quail during embryogenesis reduced cell mediated immune potential and induced a general leukocytosis in females.

Galvin, M.J.; McRee, D.I.; Hall, C.A.; Thaxton, J.P.; Parkhurst, C.R.



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.



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


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. PMID:18594834

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



Mercury levels, reproduction, and hematology in western grebes from three California Lakes, USA  

SciTech Connect

Twenty-three healthy adult western and Clark`s grebes (Aechmorphorus occidentalis and Aechmorphorus clarkii) were collected at three study sites in California, USA, in 1992: Clear Lake, Lake County; Eagle Lake, Lassen County; and Tule Lake, Siskiyou County. Liver, kidney, breast muscle, and brain were analyzed for total mercury (Hg) concentration (ppm wet weight), and blood was analyzed for various blood parameters. Clear Lake birds had greater Hg concentrations in kidney, breast muscle, and brain than birds from the other two lakes whereas liver concentrations were not statistically different. Average concentrations for Clear Lake birds were 2.74 ppm for liver, 2.06 ppm for kidney, 1.06 ppm for breast muscle, and 0.28 ppm for brain. The tissue levels of kidney, breast muscle, and brain at the other two study sites were one half the levels found at Clear Lake. These mean tissue levels were near, but below, those known to cause adverse effects. When data from all sites were merged, kidney, breast muscle, and brain concentrations are positively correlated to each other. Liver concentrations were not correlated to any other value. Brain Hg concentrations were also negatively correlated to blood potassium and blood phosphorus levels. Kidney Hg levels were positively correlated to percent blood heterophils and negatively correlated to percent eosinophils, suggesting that mercury levels might be affecting immune function. These biomarkers could not be related to any obvious ecological effects.

Elbert, R.A.; Anderson, D.W. [Univ. of California, Davis, CA (United States). Wildlife, Fish, and Conservation Biology



Myocarditis associated with reovirus in turkey poults.  


Myocarditis associated with reovirus was diagnosed in 17-day-old, male turkey poults, based on virus isolation, reverse transcriptase-polymerase chain reaction (RT-PCR), demonstration of reovirus antigen in the cytoplasm of mononuclear inflammatory cells and myocytes in the heart by immunohistochemistry (IHC), and reovirus particles in the endoplasmic reticulum of myocytes by transmission electron microscopy (TEM). Clinical signs in the poults included anorexia, growth depression, and increased mortality. Gross lesions in the six poults examined were increased pericardial fluid, mild-to-moderate dilation of right ventricles, pale-yellow myocardium, and ascites. Other lesions in a few birds included mild pulmonary edema, congestion, and pale serosa of the small intestine that had watery contents in their lumens. Microscopically, in the heart, there was mild-to-severe necrosis of myocytes and infiltration of primarily lymphocytes mixed with a few heterophils, macrophages, and occasionally, plasma cells and multinucleated giant cells. There was mild-to-moderate lymphoid depletion in the bursa of Fabricius. Reovirus was isolated from the heart of the turkey poults in chicken-embryo liver cells and was confirmed by RT-PCR, IHC, and TEM. A retrospective search of the laboratory database for cases of myocarditis associated with reovirus in turkeys revealed that this condition has occurred sporadically in California turkey flocks since 1991. This is the first documentation of myocarditis in turkey poults associated with reovirus. PMID:20095152

Shivaprasad, H L; Franca, M; Woolcock, P R; Nordhausen, R; Day, J M; Pantin-Jackwood, M



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.



Feeding reduced-protein diets to broilers subjected to hypobaric hypoxia is associated with the development of pulmonary hypertension syndrome.  


1. There are few studies on the effect of dietary protein content on pulmonary hypertension syndrome (PHS) in broiler chickens. Conflicting results from these studies prevent a clear conclusion on the effects of reduced-protein diets on development of the syndrome. 2. To obtain an understanding of the mechanisms involved, the current study, conducted at a high altitude (2100 m above sea level), evaluated the effect of three treatments that varied only according to dietary protein (CP) levels. One treatment with dietary CP advocated by National Research Council (1994) acted as a control. Two reduced-protein diets were also prepared with CP reduced 20 and 40 g/kg relative to the CP of the control, which were designated as LCPD2 and LCPD4, respectively. A total of 180 d-old male broilers (Ross 308) were randomised across 15 floor pens measuring 1.5 m(2) (12 birds per pen). Five such pens (replicates) were allotted to each dietary treatment. The protein treatments were applied from 1 to 42 d of age in which growth performance was measured and the mortality from PHS was monitored. At the end of trial (42 d), blood sampling was done and carcase characteristics were recorded. 3. Birds receiving LCPD4 gained more weight throughout the trial and had increased right ventricular weight ratio (RV:TV), relative liver weight, haematocrit, and heterophil: lymphocyte ratio at the end of the trial compared to the control. Plasma concentrations of nitric oxide (NO) and uric acid, however, were significantly lower in birds fed on LCPD4 than in those receiving the control diet. 4. Mortality from PHS was increased in birds fed the reduced-protein diets, which may have been associated with reduced concentration of plasma NO and increased haematocrit and RV:TV. In conclusion, feeding reduced-protein diets to broilers subjected to hypobaric hypoxia was associated with the development of PHS. PMID:23281761

Behrooj, N; Khajali, F; Hassanpour, H



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


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. PMID:24155914

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



Infectious antibodies in systemic lupus erythematosus patients.  


Infections can act as environmental triggers that induce or promote systemic lupus erythematosus (SLE) in genetically predisposed individuals. New technologies, developed recently, enable simultaneous assessment of multiple antibodies. Antibodies to specific infectious agents may shed light into the mechanisms of induction of SLE. The aim of this study was to investigate the prevalence of seropositivity and the titers of antibodies to bacterial, viral, and parasitic agents in SLE patients compared with non-autoimmune controls. Sera from 260 individuals (120 SLE patients and 140 controls) were tested by the BioPlex 2200 Multiplexed Immunoassay method (BioRad) for the prevalence and titers of antibodies to eight infectious agents (Epstein-Barr virus: early antigen IgG, nuclear antigen IgG, viral capsid antigen IgG and IgM, heterophile IgM; cytomegalovirus IgG and IgM; Toxoplasma gondii IgG and IgM; rubella IgG and IgM; Treponema pallidum TPr15G, TPr17G, TPr47G; herpes simplex virus type 1 and 2 IgG; hepatitis C virus and hepatitis B core antibodies. Cytomegalovirus IgM and Epstein-Barr virus early antigen IgG (but not other Epstein-Barr virus antigens) were significantly more prevalent in SLE patients than in controls. Conversely, positive titers of hepatitis B core and rubella IgG antibodies were less prevalent in the SLE patients than in controls. Other differences in titer positivity prevalence were not detected between patients and controls. The titers of the cytomegalovirus IgM, Toxoplasma IgG, Epstein-Barr virus early antigen, and viral capsid antigen IgG antibodies were significantly higher in SLE compared with controls. Our data suggest the importance of previous exposure to infectious agents in the induction and the prevention of SLE. PMID:19880558

Berkun, Y; Zandman-Goddard, G; Barzilai, O; Boaz, M; Sherer, Y; Larida, B; Blank, M; Anaya, J-M; Shoenfeld, Y



Feeding a diet contaminated with ochratoxin A for chickens at the maximum level recommended by the EU for poultry feeds (0.1 mg/kg). 1. Effects on growth and slaughter performance, haematological and serum traits.  


The European Commission Recommendation 2006/576/EC, suggests that the maximum level of Ochratoxin A (OTA) in poultry feeds should be set at 0.1 mg OTA/kg. Thirty-six one-day-old male Hubburd broiler chickens were divided into two groups, a Control (basal diet) and an Ochratoxin A (basal diet + 0.1 mg OTA/kg) group. The growth and slaughter performance traits were recorded. The liver, spleen, bursa of Fabricius and thymus weights were measured. The erythrocyte and leukocyte numbers were assayed in blood samples, and the heterophils to lymphocytes (H/L) ratio was determined. Alpha-1-acid glycoprotein (AGP), lysozyme, the total protein and the electrophoretic pattern were evaluated in serum samples. Liver enzymes (alanino aminotransferase, ALT and aspartate aminotransferase, AST) and kidney function parameters (uric acid and creatinine) were quantified. The results revealed that feeding a 0.1 mg OTA/kg contaminated diet to chicks caused a decrease in the absolute thymus weight (p < 0.05) and a lower total protein (p < 0.01), albumin (p < 0.01), alpha (p < 0.05), beta (p = 0.001) and gamma (p = 0.001) globulins serum concentration in the Ochratoxin A group. Moreover, the albumin-to-globulin (A/G) ratio of the OTA-treated animals resulted to be higher (p < 0.05). Feeding broiler chickens, a diet contaminated with the maximum level admitted by the European Commission Recommendation (0.1 mg OTA/kg), did not affect the animal performance, slaughter traits, organ weights, haematological parameters, liver enzyme or renal function parameters concentrations but had an overall immunosuppressant effect, with reduction in the thymus weight and of the total serum protein, albumin, alpha, beta and gamma globulins concentration. PMID:23639013

Pozzo, L; Salamano, G; Mellia, E; Gennero, M S; Doglione, L; Cavallarin, L; Tarantola, M; Forneris, G; Schiavone, A



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.



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



Effects of a premolt calcium and low-energy molt program on laying hen performance, egg quality, and economics.  


The objectives of this study were to evaluate and compare the effects of production, physiology, egg quality, and economics of laying hens housed in a cage system when offered a calcium premolt treatment and low-energy molt diets versus a traditional feed withdrawal (FW) treatment during and after molt. In total, 981 Hy-Line W-36 laying hens (85 wk of age) housed 3 per cage were used. Six treatments were compared in a 2 × 3 factorial design with 2 calcium premolt treatments (fine and coarse) and 3 molt diets (FW, soybean hulls, and wheat middlings). The coarse Ca was a 50:50 mix of fine (0.14-mm mean diameter) and coarse (2.27-mm mean diameter) CaCO(3), whereas the fine Ca was an all-fine CaCO(3). Both diets were formulated to contain 4.6% Ca, such that only the particle size of the CaCO(3) differed. Production parameters in experiment 1 included egg production, egg weight and mass, specific gravity, Haugh units, egg components, feed consumption and utilization, and BW. Physiological parameters in experiment 2 included ovary and oviduct weights, femur- and humerus-ash percentages, heterophil to lymphocyte ratios, plasma Ca and inorganic P concentrations, and alkaline phosphatase activity. Data were analyzed by ANOVA and P < 0.05 was significant. The fine-Ca premolt treatment was more effective than the coarse-Ca treatment at decreasing egg production during molt and increasing it postmolt, regardless of the molt diet. The FW molt diet resulted in the greatest decrease in production, but the soybean hulls diet resulted in lower production and ovary and oviduct weights during molt compared with those of the wheat middlings molt diet. Therefore, a fine-Ca premolt treatment and a low-energy molt diet, particularly soybean hulls, can be useful alternatives to a FW molt. PMID:22252340

Dickey, E R; Johnson, A K; Stalder, K J; Bregendahl, K



Functional characterization of NCAM fibronectin type III domains: demonstration of modulatory effects of the proline-rich sequence encoded by alternatively spliced exons a and AAG.  


In order to characterize the functions of the two fibronectin type III (F3) homology domains of the neural cell adhesion molecule (NCAM), we investigated the effects of two variants, expressed as fusion proteins, of the NCAM-F3 domains on attachment and spreading of NCAM-expressing fibroblasts, cerebellar cell aggregation and fiber formation, and on growth cones. The two fusion proteins were different with regard to a short proline-rich insert of six amino acids between the two F3 domains. Immobilized NCAM-F3 fusion proteins were found to mediate attachment of both transmembrane and lipid-anchored NCAM expressing fibroblasts. Also NCAM-negative cells adhered to the NCAM-F3 substratum, although to a lesser extent, implying the possibility of a heterophilic ligand to NCAM-F3 domains on the surface of fibroblasts. Cellular spreading on NCAM-F3 substratum was selectively increased in fibroblasts expressing transmembrane NCAM, and only the NCAM-F3 fusion protein lacking the proline-rich insert was able to elicit this effect. Primary cultures of mouse cerebellum were strongly inhibited with regard to formation of cellular aggregates and fibers, when incubated in the presence of either of the two NCAM-F3 fusion proteins, the fusion protein with the proline-rich insert being the more effective one. Finally, the morphology of growth cones from rat cerebellar granule cells changed significantly when grown on NCAM-F3 substrata as revealed by computer-assisted image analysis. Thus, our data indicate that the NCAM-F3 domain are involved in cell-cell adhesion, and that insertion of the proline-rich sequence has a modulatory effect on NCAM-F3 domain functions. PMID:8915894

Kasper, C; Stahlhut, M; Berezin, V; Maar, T E; Edvardsen, K; Kiselyov, V V; Soroka, V; Bock, E



Diversity in specificity, abundance, and composition of anti-Neu5Gc antibodies in normal humans: Potential implications for disease  

PubMed Central

Human heterophile antibodies that agglutinate animal erythrocytes are known to detect the nonhuman sialic acid N-glycolylneuraminic acid (Neu5Gc). This monosaccharide cannot by itself fill the binding site (paratope) of an antibody and can also be modified and presented in various linkages, on diverse underlying glycans. Thus, we hypothesized that the human anti-Neu5Gc antibody response is diverse and polyclonal. Here, we use a novel set of natural and chemoenzymatically synthesized glycans to show that normal humans have an abundant and diverse spectrum of such anti-Neu5Gc antibodies, directed against a variety of Neu5Gc-containing epitopes. High sensitivity and specificity assays were achieved by using N-acetylneuraminic acid (Neu5Ac)-containing probes (differing from Neu5Gc by one less oxygen atom) as optimal background controls. The commonest anti-Neu5Gc antibodies are of the IgG class. Moreover, the range of reactivity and Ig classes of antibodies vary greatly amongst normal humans, with some individuals having remarkably large amounts, even surpassing levels of some well-known natural blood group and xenoreactive antibodies. We purified these anti-Neu5Gc antibodies from individual human sera using a newly developed affinity method and showed that they bind to wild-type but not Neu5Gc-deficient mouse tissues. Moreover, they bind back to human carcinomas that have accumulated Neu5Gc in vivo. As dietary Neu5Gc is primarily found in red meat and milk products, we suggest that this ongoing antigen-antibody reaction may generate chronic inflammation, possibly contributing to the high frequency of diet-related carcinomas and other diseases in humans.

Padler-Karavani, Vered; Yu, Hai; Cao, Hongzhi; Chokhawala, Harshal; Karp, Felix; Varki, Nissi; Chen, Xi; Varki, Ajit



Influence of Salmonella enterica serovar Typhimurium infection on intestinal goblet cells and villous morphology in broiler chicks.  


Live broiler chickens are important in the transmission of Salmonella to humans. Reducing Salmonella levels in the intestine of broiler chickens, in part, requires understanding of the interactions between Salmonella and the intestinal barriers that represent the first line of defense. Such barriers include the mucus layer (composed of mucins secreted by goblet cells) and the underlying epithelium. Two experiments were conducted to evaluate the effect of Salmonella Typhimurium infection on intestinal goblet cell dynamics (density and size) and villous morphology in broiler chicks. In Experiment 1, broiler chicks were either challenged with sterile media (control treatment) or orally given 7.4 x 10(7) colony-forming units (CFU) at 3 days of age (termed the CST treatment). Treatments were similar in Experiment 2, except that chicks in the CST treatment were challenged with 7.8 x 10(6) CFU at 4 days of age. Duration of each experiment was 14 days. At 7 days postchallenge (PC) in Experiment 1, jejunal tissue sections were collected, formalin-fixed, and routinely processed for histologic measurement of villous morphometric indices. In Experiment 2, at 10 days PC, jejunal tissue sections were collected and processed for histologic determination of goblet cell numbers and size, in addition to villous measurements. Results showed that Salmonella Typhimurium infection increased goblet cell density, reduced villous surface area, increased the incidence of epithelial exfoliation, and increased the incidence of heterophil influx into the lamina propria (P < 0.05). It was concluded that Salmonella Typhimurium infection impacts goblet cell biology and exerts morphopathologic changes in the jejunum of broiler chicks. PMID:20608528

Fasina, Y O; Hoerr, F J; McKee, S R; Conner, D E



ALCAM regulates mediolateral retinotopic mapping in the superior colliculus.  


ALCAM [activated leukocyte cell adhesion molecule (BEN/SC-1/DM-GRASP)] is a transmembrane recognition molecule of the Ig superfamily (IgSF) containing five Ig domains (two V-type, three C2-type). Although broadly expressed in the nervous and immune systems, few of its developmental functions have been elucidated. Because ALCAM has been suggested to interact with the IgSF adhesion molecule L1, a determinant of retinocollicular mapping, we hypothesized that ALCAM might direct topographic targeting to the superior colliculus (SC) by serving as a substrate within the SC for L1 on incoming retinal ganglion cell (RGC) axons. ALCAM was expressed in the SC during RGC axon targeting and on RGC axons as they formed the optic nerve; however, it was downregulated distally on RGC axons as they entered the SC. Axon tracing with DiI revealed pronounced mistargeting of RGC axons from the temporal retina half of ALCAM null mice to abnormally lateral sites in the contralateral SC, in which these axons formed multiple ectopic termination zones. ALCAM null mutant axons were specifically compromised in medial orientation of interstitial branches, which is known to require the ankyrin binding function of L1. As a substrate, ALCAM-Fc protein promoted L1-dependent attachment of acutely dissociated retinal cells and an L1-expressing, ALCAM-negative cell line, consistent with an ALCAM-L1 heterophilic molecular interaction. Together, these results suggest a model in which ALCAM in the SC interacts with L1 on RGC axons to promote medial extension of RGC axon branches important for mediolateral axon targeting in the formation of retinocollicular maps. PMID:20016077

Buhusi, Mona; Demyanenko, Galina P; Jannie, Karry M; Dalal, Jasbir; Darnell, Eli P B; Weiner, Joshua A; Maness, Patricia F



Immuno-phenotypic and functional characterization of rabbit pulmonary intravascular macrophages.  


Pulmonary intravascular macrophages (PIMs) are present in species such as cattle, sheep and horse and promote acute lung inflammation (ALI). Rabbits are often used as a model of ALI but there is controversy about the presence of PIMs in these species. Rabbits were treated with 10 mg/kg of gadolinium chloride intravenously (GC; n?=?6) or saline (n?=?6) followed by euthanasia at 48 h post-treatment to determine the presence of PIMs. In a subsequent study, rabbits were pre-treated with GC or 0.9 % saline followed by 100 ?g/kg of E. coli lipopolysaccharide intravenously 48 h later. Rabbits were euthanized 24 h post-LPS treatment. Light and electron microscopy showed that PIMs attached to the capillary endothelium and were positive for RAM-11 anti-macrophage antibody. While GC treatment induced apoptotic PIMs, there was no difference in the PIM number between control and GC-treated rabbits. Rabbits administered with LPS were 3.5 times more likely to die before the end of the 24-h period than those pre-treated with GC. Lung heterophil accumulation and IL-1?, TNF? and IL-6 mRNA expression were significantly higher in rabbits administered with LPS compared to those administered with GC before the LPS injection. PIMs from the LPS-treated rabbits were positive for TNF?. Lung, BAL and serum IL-8 and MCP-1 expression was not different between LPS rabbits with or without pre-treatment with GC. We conclude that rabbit lungs contain PIMs and that their depletion reduces endotoxin-induced lung inflammation. The presence of PIMs in rabbit lungs may need to be considered while using rabbit to model acute lung injury. PMID:23073615

Duke-Novakovski, Tanya; Singh-Suri, Sarabjeet; Kajikawa, Osamu; Caldwell, Sarah; Charavaryamath, Chandarshekhar; Singh, Baljit



Re-evaluation of chicken CXCR1 determines the true gene structure: CXCLi1 (K60) and CXCLi2 (CAF/interleukin-8) are ligands for this receptor.  


The original report of chicken CXCR1 (Li, Q. J., Lu, S., Ye, R. D., and Martins-Green, M. (2000) Gene (Amst.) 257, 307-317) described it as a single exon gene, with two isoforms (differing in their start codon). In comparison with mammalian CXCR1, the reported chicken CXCR1 was longer at both the NH(2) and COOH termini, and it lacked the conserved (C/S)CXNP motif present in the last transmembrane region of all known chemokine receptors. A re-evaluation of chicken CXCR1, comparing known expressed sequence tags with the chicken genome sequence, suggested that the gene contains two exons. We isolated a cDNA corresponding to our prediction, which was significantly different in sequence to the reported CXCR1. In particular, there were three frameshifts in our sequence, compared with the reported sequence, that restored higher identity in the COOH-terminal half of the protein to mammalian CXCR1 (61% total amino acid identity compared with 52% for the reported CXCR1), restored the (C/S)CXNP motif, and gave a predicted protein of the same length as mammalian CXCR1. In human, CXCR1 is the receptor for CXCL8. In the chicken, there are two syntenic genes, CXCLi1 and CXCLi2, which look equally like orthologues of human CXCL8. We demonstrate that both of these chemokines are ligands for chicken CXCR1. We also demonstrate that heterophils express chicken CXCR1 and that the receptor is Galpha(i) protein-linked. PMID:18417470

Poh, Tuang Yeow; Pease, James; Young, John R; Bumstead, Nat; Kaiser, Pete



Hematologic reference values for clinically healthy captive golden conures (Guaruba guarouba).  


Golden conures or ararajubas (Guaruba guarouba) are endangered parrots endemic to the Brazilian Amazon forest. Body mass, blood cell counts, and total plasma protein were determined for 70 clinically healthy golden conures captive at zoologic parks and private breeder facilities in Brazil. Hematologic results (mean +/- SD) were: Erythrocytes 3.6 +/- 0.5 x 10(6) cells/mm3, hemoglobin 12.8 +/- 1.4 g/dl, packed cell volume 46 +/- 3.8%, mean corpuscular volume 132 +/- 20 fl, mean corpuscular hemoglobin (MCH) 36 +/- 5.7 pg, mean corpuscular hemoglobin concentration (MCHC) 28 +/- 3.5%, thrombocytes 26.3 +/- 9.3 x 10(3) cells/mm3, leukocytes 11.9 +/- 4.5 x 10(3) cells/mm3, heterophils 6284 +/- 2715 cells/mm3, lymphocytes 5473 +/- 2408 cells/ mm3, monocytes 113 +/- 162 cells/mm3, eosinophils 10 +/- 42 cells/mm3, basophils 27 +/- 64 cells/mm3. Body mass was 254 +/- 24.9 g and total plasma protein (TPP) was 3.54 +/- 0.58 g/dl. No statistical differences were observed between genders within age groups. Differences between juveniles (J) and adults (A) were identified for TPP (J < A), MCH (J > A), and MCHC (J > A). These results provide reliable reference values for the clinical interpretation of hematologic results for the species. Hematology may be an important tool for population health investigations on free-ranging golden conure populations and will also be essential to survey the health of release candidates in future reintroduction programs. PMID:23397841

Prioste, Fabíola Eloisa Setim; Zwarg, Ticiana; Teixeira, Rodrigo Hidalgo; Vanstreels, Ralph Eric Thijl; Rocha, Arnaldo; Matushima, Eliana Reiko



Pathology and pathophysiology of inhalational anthrax in a guinea pig model.  


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 × 10(4) 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. PMID:23357384

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-Biérinx, Laurence; Park, Sukjoon; Ionin, Boris; Skiadopoulos, Mario H



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



Mechanism of Fc? Receptor-Mediated Trogocytosis-Based False-Positive Results in Flow Cytometry  

PubMed Central

The whole blood erythrocyte lysis method is the most common protocol of sample preparation for flow cytometry (FCM). Although this method has many virtues, our recent study has demonstrated false-positive results when surface markers of monocytes were examined by this method due to the phenomenon called Fc? receptor (Fc?R)-mediated trogocytosis. In the present study, similar Fc?R-mediated trogocytosis-based false-positive results have been demonstrated when granulocytes were focused on instead of monocytes. These findings indicated that not only monocytes but also granulocytes, the largest population with Fc?R expression in peripheral blood, could perform Fc?R-mediated trogocytosis. Since the capacity of Fc?R-mediated trogocytosis was different among blood samples, identification of factors that could regulate the occurrence of Fc?R-mediated trogocytosis should be important for the quality control of FCM. Our studies have suggested that such factors are present in the serum. In order to identify the serum factors, we employed the in vitro model of Fc?R-mediated trogocytosis using granulocytes. Investigation with this model determined the serum factors as heat-labile molecules with molecular weight of more than 100 kDa. Complements in the classical pathway were initially assumed as candidates; however, the C1 inhibitor did not yield an obvious influence on Fc?R-mediated trogocytosis. On the other hand, although immunoglobulin ought to be resistant to heat inactivation, the inhibitor of human anti-mouse antibodies (HAMA) effectively blocked Fc?R-mediated trogocytosis. Moreover, the inhibition rates were significantly higher in HAMAhigh serum than HAMAlow serum. The collective findings suggested the involvement of heterophilic antibodies such as HAMA in the mechanism of false-positive results in FCM due to Fc?R-mediated trogocytosis.

Masuda, Sakiko; Iwasaki, Sari; Tomaru, Utano; Sato, Juri; Kawakami, Ai; Ichijo, Kana; Sogo, Sayuri; Baba, Tomohisa; Katsumata, Kazuaki; Kasahara, Masanori; Ishizu, Akihiro



The neural cell adhesion molecule-derived peptide FGL facilitates long-term plasticity in the dentate gyrus in vivo.  


The neural cell adhesion molecule (NCAM) is known to play a role in developmental and structural processes but also in synaptic plasticity and memory of the adult animal. Recently, FGL, a NCAM mimetic peptide that binds to the Fibroblast Growth Factor Receptor 1 (FGFR-1), has been shown to have a beneficial impact on normal memory functioning, as well as to rescue some pathological cognitive impairments. Whether its facilitating impact may be mediated through promoting neuronal plasticity is not known. The present study was therefore designed to test whether FGL modulates the induction and maintenance of synaptic plasticity in the dentate gyrus (DG) in vivo. For this, we first assessed the effect of the FGL peptide on synaptic functions at perforant path-dentate gyrus synapses in the anesthetized rat. FGL, or its control inactive peptide, was injected locally 60 min before applying high-frequency stimulation (HFS) to the medial perforant path. The results suggest that although FGL did not alter basal synaptic transmission, it facilitated both the induction and maintenance of LTP. Interestingly, FGL also modified the heterosynaptic plasticity observed at the neighboring lateral perforant path synapses. The second series of experiments, using FGL intracerebroventricular infusion in the awake animal, confirmed its facilitating effect on LTP for up to 24 h. Our data also suggest that FGL could alter neurogenesis associated with LTP. In sum, these results show for the first time that enhancing NCAM functions by mimicking its heterophilic interaction with FGFR facilitates hippocampal synaptic plasticity in the awake, freely moving animal. PMID:21508096

Dallérac, Glenn; Zerwas, Meike; Novikova, Tatiana; Callu, Delphine; Leblanc-Veyrac, Pascale; Bock, Elisabeth; Berezin, Vladimir; Rampon, Claire; Doyère, Valérie



Human cytomegalovirus UL7, a homologue of the SLAM-family receptor CD229, impairs cytokine production.  


Human cytomegalovirus (HCMV), the ?-herpesvirus prototype, has evolved a wide spectrum of mechanisms to counteract host immunity. Among them, HCMV uses cellular captured genes encoding molecules capable of interfering with the original host function or of fulfilling new immunomodulatory tasks. Here, we report on UL7, a novel HCMV heavily glycosylated transmembrane protein, containing an Ig-like domain that exhibits remarkable amino acid similarity to CD229, a cell-surface molecule of the signalling lymphocyte-activation molecule (SLAM) family involved in leukocyte activation. The UL7 Ig-like domain, which is well-preserved in all HCMV strains, structurally resembles the SLAM-family N-terminal Ig-variable domain responsible for the homophilic and heterophilic interactions that trigger signalling. UL7 is transcribed with early-late kinetics during the lytic infectious cycle. Using a mAb generated against the viral protein, we show that it is constitutively shed, through its mucine-like stalk, from the cell-surface. Production of soluble UL7 is enhanced by PMA and reduced by a broad-spectrum metalloproteinase inhibitor. Although UL7 does not hold the ability to interact with CD229 or other SLAM-family members, it shares with them the capacity to mediate adhesion to leukocytes, specifically to monocyte-derived DCs. Furthermore, we demonstrate that UL7 expression attenuates the production of proinflammatory cytokines TNF, IL-8 and IL-6 in DCs and myeloid cell lines. Thus, the ability of UL7 to interfere with cellular proinflammatory responses may contribute to viral persistence. These results enhance our understanding of those HCMV-encoded molecules involved in sustaining the balance between HCMV and the host immune system. PMID:21670740

Engel, Pablo; Pérez-Carmona, Natàlia; Albà, M Mar; Robertson, Kevin; Ghazal, Peter; Angulo, Ana



Cellular Localization of Nectin-1 and Glycoprotein D during Herpes Simplex Virus Infection  

PubMed Central

During viral entry, herpes simplex virus (HSV) glycoprotein D (gD) interacts with a specific cellular receptor such as nectin-1 (PRR1/HveC/CD111) or the herpesvirus entry mediator A (HVEM/HveA). Nectin-1 is involved in cell-to-cell adhesion. It is located at adherens junctions, where it bridges cells through homophilic or heterophilic interactions with other nectins. Binding of HSV gD prevents nectin-1-mediated cell aggregation. Since HSV gD affects the natural function of nectin-1, we further investigated the effects of gD expression on nectin-1 during HSV infection or in transfected cells. We also studied the importance of the interaction between nectin-1 and the cytoplasmic protein afadin for HSV entry and spread as well as the effects of infection on this interaction. In these investigations, we used a panel of cells expressing nectin-1 or nectin-1-green fluorescent protein fusions as the only mediators of HSV entry. During HSV infection, nectin-1 localization at adherens junction was dramatically altered in a manner dependent on gD expression. Nectin-1 and gD colocalized at cell contact areas between infected and noninfected cells and at the edges of plaques. This specific accumulation of gD at junctions was driven by expression of nectin-1 in trans on the surface of adjacent cells. Reciprocally, nectin-1 was maintained at junctions by the trans expression of gD in the absence of a cellular natural ligand. Our observations indicate that newly synthesized gD substitutes for nectin-1 of infected cells at junctions with noninfected cells. We propose that gD attracts and maintains the receptor at junctions where it can be used for virus spread.

Krummenacher, Claude; Baribaud, Isabelle; Eisenberg, Roselyn J.; Cohen, Gary H.



Vitamin D measurement in the intensive care unit: methodology, clinical relevance and interpretation of a random value.  


Vitamin D deficiency, as measured by a random level of 25-hydroxyvitamin D is very prevalent in critically ill patients admitted to the ICU and is associated with adverse outcomes. Both 25(OH)vitamin D and 1?,25(OH)2D3 are difficult to analyse because of their lipophilic nature, affinity for VDBP and small concentrations. Also, the various tests used to estimate vitamin D levels show significant inter- and intra-assay variability, which significantly affect the veracity of the results obtained and confound their interpretation. The two main types of assays include those that directly estimate vitamin D levels (HPLC, LC-MS/MS) and competitive binding assays (RIA, EIA). The former methods require skilled operators, with prolonged assay times and increased cost, whereas the latter are cheaper and easy to perform, but with decreased accuracy. The direct assays are not affected by lipophilic substances in plasma and heterophile antibodies, but may overestimate vitamin D levels by measuring the 3-epimers. These problems can be eliminated by adequate standardization of the test using SRMs provided by NIST, as well as participating in proficiency schemes like DEQAS. It is therefore important to consider the test employed as well as laboratory quality control, while interpreting vitamin D results. A single random measurement may not be reflective of the vitamin D status in ICU patients because of changes with fluid administration, and intra-day variation in 25-hydroxyvitamin D levels. 1?,25(OH)2D3 may behave differently to 25-hydroxyvitamin D, both in plasma and at tissue level, in inflammatory states. Measurement of tissue 1?,25(OH)2D3 levels may provide the true estimate of vitamin D activity. PMID:23782207

Krishnan, Anand; Venkatesh, Bala



Characterization of neutrophil extracellular traps in cats naturally infected with feline leukemia virus.  


Feline leukemia virus (FeLV), a common, naturally occurring gammaretrovirus in domestic cats, is associated with degenerative diseases of the haematopoietic system, immunodeficiency and neoplasia. FeLV infection causes an important suppression of neutrophil function, leading to opportunistic infections. Recently, a new microbicidal mechanism named NETosis was described in human, bovine and fish neutrophils, as well as in chicken heterophils. The purpose of the present study was to characterize NETosis in feline neutrophils, as well as to evaluate neutrophil function in FeLV naturally infected symptomatic and asymptomatic cats through the phagocytosis process, release of neutrophil extracellular traps (NETs) and myeloperoxidase (MPO) activity. The results showed that feline neutrophils stimulated with protozoa parasites released structures comprising DNA and histones, which were characterized as NETs by immunofluorescence. Quantification of NETs after neutrophil stimulation showed a significant increase in NET release by neutrophils from FeLV(-) and FeLV(+) asymptomatic cats compared with FeLV(+) symptomatic cats. Moreover, the number of released NETs and MPO activity in unstimulated neutrophils of FeLV(+) symptomatic cats were higher than those in unstimulated neutrophils from FeLV(-) and FeLV(+) asymptomatic cats. This study reports, for the first time, NET release by feline neutrophils, along with the fact that NET induction may be modulated by a viral infection. The results indicate that the NET mechanism appears to be overactivated in FeLV(+) cats and that this feature could be considered a marker of disease progression in FeLV infection. PMID:19793908

Wardini, Amanda B; Guimarães-Costa, Anderson B; Nascimento, Michelle T C; Nadaes, Natalia R; Danelli, Maria G M; Mazur, Carlos; Benjamim, Claudia F; Saraiva, Elvira M; Pinto-da-Silva, Lucia H



Experimental challenge of mallards (Anas platyrhynchos) with Brachyspira hyodysenteriae and "Brachyspira suanatina" isolated from pigs and mallards.  


Brachyspira hyodysenteriae, the aetiological agent of swine dysentery, and a recently proposed and closely related enteropathogenic spirochaete "Brachyspira suanatina", originally isolated from pigs or mallards (Anas platyrhynchos), were used to inoculate week-old mallard ducklings orally or cloacally. The colonization rate, clinical outcome, faecal dry matter content, blood leucocyte counts and gross, microscopical and electron microscopical features 14-16 days post-inoculation were investigated at necropsy examination. Strains of "B. suanatina" of pig and mallard origin and B. hyodysenteriae of mallard origin colonized the ducklings by oral inoculation, and colonization was also established by cloacal inoculation with a "B. suanatina" strain of mallard origin. The porcine reference strain of B. hyodysenteriae (B204) failed to colonize the birds. Unchallenged contact birds in one of the challenge groups were readily colonized by a strain of "B. suanatina" of mallard origin. The proportion of colonized birds differed significantly between the challenge groups (P < 0.0001). For each challenge group, the inoculum and a randomly selected subset of recovered isolates had an identical biochemical profile and banding pattern by randomly amplified polymorphic DNA (RAPD) analysis. None of the birds developed clinical signs of gastrointestinal disease during the trial. The faecal dry weight contents, body weights and total leucocyte and heterophil counts did not differ between the various groups of birds. At the microscopical and electron microscopical levels, the caecal mucosa in some of the Brachyspira culture-positive birds had sharply demarcated epithelial cell changes and there were features of irreversible cell damage in crypt necks coinciding with spirochaetal infiltration of the mucosa. The crypts in Brachyspira culture-positive birds were deeper than in culture-negative birds (median: 237 microm and 218 microm, respectively, P = 0.019). This challenge model was well suited for use in mallards and consistent with previous findings that strongly haemolytic Brachyspira spp. may cross the species barrier between pigs and birds. PMID:19640549

Jansson, D S; Råsbäck, T; Fellström, C; Feinstein, R



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



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



Marek's disease virus infection in the eye: chronological study of the lesions, virus replication, and vaccine-induced protection.  


Marek's disease virus (MDV) infection in the eye was studied chronologically after inoculating 1-day-old chickens with a very virulent MDV strain, Md5. The ocular lesions could be classified as early lesions (6-11 days postinoculation [dpi]) and late lesions (26 and 56 dpi), based upon the location and severity of the lesions. The early lesions involved iris, ciliary body, and choroid layer, and were characterized by endothelial cell hypertrophy, vasculitis, and infiltration of lymphocytes (mainly CD8+), plasma cells, macrophages, and heterophils. Expression of early MDV-antigen pp38 in the cells infiltrating choroid layer was detected as early as 11 dpi. Late lesions consisted of severe lymphohistiocytic uveitis, keratitis, pectenitis, vitreitis, retinitis, and segmental to diffuse retinal necrosis. Cell infiltration included macrophages, granulocytes, plasma cells, and both CD4+ and CD8+ cells of various sizes. Expression of early MDV-antigen pp38 was readily found within the retina, uveal tract, and corneal epithelium. No expression of late-antigen gB or oncoprotein meq was detected in any of the eyes examined. A second experiment was conducted to study the effect of vaccination on the development of ocular lesions. Both HVT and CVI988 were able to protect against the development of early ocular lesions in chickens infected with very virulent plus strain MDV 648A. However, only CVI988 conferred complete protection against the development of late ocular lesions. HVT conferred partial protection, as it reduced the frequency and severity of the late ocular lesions. These results enhance our understanding of the nature and pattern of MDV infection in the eye. PMID:19166047

Pandiri, Arun K R; Cortes, Aneg L; Lee, Lucy F; Gimeno, I M



[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


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



Comparisons of Highly Virulent H5N1 Influenza A Viruses Isolated from Humans and Chickens from Hong Kong  

PubMed Central

Genes of an influenza A (H5N1) virus from a human in Hong Kong isolated in May 1997 were sequenced and found to be all avian-like (K. Subbarao et al., Science 279:393–395, 1998). Gene sequences of this human isolate were compared to those of a highly pathogenic chicken H5N1 influenza virus isolated from Hong Kong in April 1997. Sequence comparisons of all eight RNA segments from the two viruses show greater than 99% sequence identity between them. However, neither isolate’s gene sequence was closely (>95% sequence identity) related to any other gene sequences found in the GenBank database. Phylogenetic analysis demonstrated that the nucleotide sequences of at least four of the eight RNA segments clustered with Eurasian origin avian influenza viruses. The hemagglutinin gene phylogenetic analysis also included the sequences from an additional three human and two chicken H5N1 virus isolates from Hong Kong, and the isolates separated into two closely related groups. However, no single amino acid change separated the chicken origin and human origin isolates, but they all contained multiple basic amino acids at the hemagglutinin cleavage site, which is associated with a highly pathogenic phenotype in poultry. In experimental intravenous inoculation studies with chickens, all seven viruses were highly pathogenic, killing most birds within 24 h. All infected chickens had virtually identical pathologic lesions, including moderate to severe diffuse edema and interstitial pneumonitis. Viral nucleoprotein was most frequently demonstrated in vascular endothelium, macrophages, heterophils, and cardiac myocytes. Asphyxiation from pulmonary edema and generalized cardiovascular collapse were the most likely pathogenic mechanisms responsible for illness and death. In summary, a small number of changes in hemagglutinin gene sequences defined two closely related subgroups, with both subgroups having human and chicken members, among the seven viruses examined from Hong Kong, and all seven viruses were highly pathogenic in chickens and caused similar lesions in experimental inoculations.

Suarez, David L.; Perdue, Michael L.; Cox, Nancy; Rowe, Thomas; Bender, Catherine; Huang, Jing; Swayne, David E.



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?

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



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

PubMed Central

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.

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



Diarrhea induced by Treponema hyodysenteriae: a young chick cecal model for swine dysentery.  

PubMed Central

The experimental infection of 25 young chicks with Treponema hyodysenteriae was carried out. Treponemes were isolated from 21 of 25 chicks on day 21 after inoculation. The ceca of chicks infected with treponemes were atrophied. The lumen was filled with a white watery fluid instead of digested feed. In some infected chicks, a cecal core was observed with the fluid in the cecum. The cecal core was grayish, hard, and rod shaped. It consisted of eroded cells and debris of treponemes and resembled the pseudomembrane. Bloody mucus was also observed in one chick. The thickness of the mucosae in 17 of 25 chicks were markedly increased. The histological changes were classified into two types. In the case of regressive changes of epithelial cells which mean severe erosion, the laminae propriae were exposed. Hemorrhage, edema, and heterophil infiltration in the laminae propriae were also confirmed. Numerous treponemes were observed within the edematous area under the remaining epithelia and also invaded the epithelial cells and laminae propriae. In the other case, progressive changes, that is, hyperplasia of mucosal epithelial cells and elongation of the crypt, were observed. The epithelia consisted mainly of cuboidal basophilic cells, mitotic cells, and goblet cells. The mitotic cells increased in number and were also observed near the superficial luminal surface of the ceca. Mucous goblet cells were also considerably increased in number. The erosion of superficial luminal epithelial cells was not so severe, but edema in laminae propriae was frequently observed. Electron-microscopic observation demonstrated that the basophilic epithelial cells were polyribosome rich, mitochondria poor, and lipid droplet poor. Furthermore, tonofibril-like structures under the terminal web in cytoplasms were lost, and numerous membrane-bound vesicles at the terminal web with free ribosomes were observed. In places, a number of vesicles were observed between microvilli, and some vesicles were released from the apical cytoplasm of epithelial cells. Microvilli also became scarce and irregular. The junctional complexes between cuboidal cells became unclear, the interdigitations between cuboidal cells became loose, and the intercellular spaces were widened. In conclusion, the appearance of numerous membrane-bound vesicles at the terminal web and the dilated intercellular space indicates increased secretion of water and electrolytes, and the abnormal proliferation of such immature epithelial cells indicates impaired absorption. On the basis of our findings, we can say that diarrhea induced by T. hyodysenteriae can be developed by synergism between impaired absorption and increased secretion by intestinal mucosae with hyperplastic immature epithelial cells.(ABSTRACT TRUNCATED AT 400 WORDS) Images

Sueyoshi, M; Adachi, Y



Deletion analysis of two tandemly arranged virulence genes in myxoma virus, M11L and myxoma growth factor.  

PubMed Central

Myxoma virus (MYX) is a leporipoxvirus of rabbits that induces a lethal syndrome characterized by disseminated tumorlike lesions, generalized immunosuppression, and secondary gram-negative bacterial infection. A MYX deletion mutant (vMYX-GF- delta M11L) was constructed to remove the entire myxoma growth factor (MGF) coding sequence and that for the C-terminal five amino acids of the partially overlapping upstream gene, M11L. Unexpectedly, this deletion completely abrogates the capacity of MYX to cause the characteristic disease symptoms of myxomatosis. Upon inoculation of rabbits with vMYX-GF- delta M11L, recipient animals developed only a benign, localized nodule reminiscent of a Shope fibroma virus-induced tumor in which a single primary lesion appeared at the site of injection and then completely regressed within 14 days, leaving the animals resistant to challenge with wild-type MYX. No evidence of the purulent conjunctivitis and rhinitis that always accompany wild-type MYX infection was observed. To ascertain whether the attenuation observed in vMYX-GF- delta M11L was due to a combined effect of the MGF deletion and alteration of the upstream M11L gene, two additional MYX recombinants were constructed: an MGF- virus (vMYX-GF-) containing an intact M11L gene and an M11L- virus (vMYX-M11L-) containing an intact MGF gene. Infection with vMYX-GF- resulted in moderated symptoms of myxomatosis, but all clinical stages of the disease were still detectable. In contrast, disruption of M11L alone dramatically reduced the virus virulence, resulting in a nonlethal syndrome whose clinical course was nevertheless distinct from that of vMYX-GF- delta M11L. Upon inoculation with vMYX-M11L-, rabbits developed primary and secondary tumors which were larger and more circumscribed than those of wild-type MYX recipients. Whereas wild-type MYX infection always includes severe, purulent conjunctivitis and rhinitis, vMYX-M11L- recipients remained healthy and displayed only minimal signs of respiratory distress. By about 30 days after infection, the tumors induced by vMYX-M11L- had completely regressed and these animals were immune to challenge with wild-type MYX. Histological analysis indicated that tumors induced by vMYX-M11L- are much more heavily infiltrated with macrophages and heterophils and that the sites of viral replication are more edematous and necrotic than those of wild-type infection, suggesting that the host was able to mount a more vigorous inflammatory response to vMYX-M11L- infection.(ABSTRACT TRUNCATED AT 400 WORDS) Images

Opgenorth, A; Graham, K; Nation, N; Strayer, D; McFadden, G



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

NASA Astrophysics Data System (ADS)

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). Body weight and feed intake were not influenced significantly by dietary vitamin E and Se (P > 0.05), whereas feed conversion was improved significantly by 125 mg/kg vitamin E (P < 0.05). The liver and lymphoid organ weights as well as IgM and IgG, antibody titers for primary and secondary antibody responses to SRBC were reduced significantly under HS (P < 0.05). Heat stress also resulted in a significant increase in H/L ratio (P < 0.05). Dietary vitamin E resulted in improvement of primary and secondary antibody responses both in TN and HS broilers (P < 0.05). The HS birds also showed an improved antibody titer in secondary response with high concentration of Se (P < 0.05). Vitamin E and Se had interactive effects on anti-SRBC titers; however, no consistent differences were found between dietary levels during the study. The H/L ratio decreased by feeding vitamin E at both levels either under HS or TN conditions (P < 0.05). The serum concentrations of glucose, triglycerides, total cholesterol, and LDL-cholesterol were increased but serum HDL-cholesterol decreased in HS broilers (P < 0.05).

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



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



Non-clustered protocadherin  

PubMed Central

The cadherin family is classified into classical cadherins, desmosomal cadherins and protocadherins (PCDHs). Genomic structures distinguish between PCDHs and other cadherins, and between clustered and non-clustered PCDHs. The phylogenetic analysis with full sequences of non-clustered PCDHs enabled them to be further classified into three subgroups: ?1 (PCDH1, PCDH7, PCDH9, PCDH11 and PCDH20), ?2 (PCDH8, PCDH10, PCDH12, PCDH17, PCDH18 and PCDH19) and ? (PCDH15, PCDH16, PCDH21 and MUCDHL). ?-PCDH members except PCDH21 have either higher or lower numbers of cadherin repeats than those of other PCDHs. Non-clustered PCDHs are expressed predominantly in the nervous system and have spatiotemporally diverse expression patterns. Especially, the region-specific expressions of non-clustered PCDHs have been observed in cortical area of early postnatal stage and in caudate putaman and/or hippocampal formation of mature brains, suggesting that non-clustered PCDHs play roles in the circuit formation and maintenance. The non-clustered PCDHs appear to have homophilic/heterophilic cell-cell adhesion properties, and each member has diverse cell signaling partnership distinct from those of other members (PCDH7/TAF1; PCDH8/TAO2?; PCDH10/Nap1; PCDH11/?-catenin; PCDH18/mDab1). Furthermore, each PCDH has several isoforms with differential cytoplasmic sequences, suggesting that one PCDH isoform could activate intracellular signaling differential from other isoforms. These facts suggest that non-clustered PCDHs play roles as a mediator of a regulator of other molecules as well as cell-cell adhesion. Furthermore, some non-clustered PCDHs have been considered to be involved in neuronal diseases such as autism-spectrum disorders, schizophrenia and female-limited epilepsy and cognitive impairment, suggesting that they play multiple, tightly regulated roles in normal brain function. In addition, some non-clustered PCDHs have been suggested as candidate tumor suppressor genes in several tissues. Although molecular adhesive and regulatory properties of some PCDHs began to be unveiled, the endeavor to understand the molecular mechanism of non-clustered PCDH is still in its infancy and requires future study.

Kim, Soo-Young; Yasuda, Shin; Tanaka, Hidekazu



Anti-carbohydrate antibodies of normal sera: findings, surprises and challenges.  


We have used microchip format glycan array to characterize the individual carbohydrate recognition patterns by antibodies (Ab) in sera of 106 healthy donors. The glycan library included blood group antigens and other most frequent terminal oligosaccharides and their cores of mammalian N- and O-linked glycoproteins and glycolipids, tumor-associated carbohydrate antigens, and common components of bacterial/pathogenic polysaccharides and lipopolysaccharides, totally 205 glycans. The serum Ab interacted with at least 50 normal human glyco-motifs. Apart from expected blood group-, xeno- (heterophil) and infection-related binding activities, we observed a number of new and unexpected features. The surprising, relatively high antibody binding was found to the blood group P(1) and P(k) trisaccharides and H(type 2) trisaccharide. Novel and very high binding activities have been observed towards Galbeta1-3GlcNAc (Le(C)) related glycans, especially 3'-O-Su-Le(C), and towards 4'-O-sulfated lactosamine. Relatively high and uniform Ab binding to GalNAcalpha1-3Gal disaccharide demonstrated absence of correlation with fucosylated blood group A GalNAcalpha1-3(Fucalpha1-2)Gal antigen-similarly to well known relationship between Galalpha1-3Gal and true, fucosylated blood group B Galalpha1-3(Fucalpha1-2)Gal antigen. The binding intensity to Galalpha1-3Galbeta1-4GlcNAc xenoantigen was shown to be rather modest. Absence or very low Ab binding was found against oligosialic acid, sialooligosaccharides except SiaT(n), type 2 backbone glycans such as Le(y), and biantennary N-chain as well as its truncated forms, i.e. without terminal Sia, SiaGal, and SiaGalGlcNAc motifs. We have also found that Ab are capable of recognizing the short inner core typical for glycolipids (-Galbeta1-4Glc) and glycoproteins (-GalNAcalpha) as a fragment of bigger glycans. PMID:19608278

Huflejt, Margaret E; Vuskovic, Marko; Vasiliu, Daniela; Xu, Hongyu; Obukhova, Polina; Shilova, Nadezhda; Tuzikov, Alexander; Galanina, Oxana; Arun, Banu; Lu, Karen; Bovin, Nicolai



The effects of a short-term molt method using cassava meal, broken rice, or corn on ovarian regression, bone integrity, and postmolt egg production and quality in older (95 week) laying hens.  


This study was conducted to determine the ability of different molt diets to induce molt in 95-wk-old hens. The hens were randomly assigned to 4 treatments of 60 birds each: 1) molted by full feeding with broken rice (BROK), 2) corn (CRN), 3) cassava (CASS), or 4) nonmolted control (CONT) for 14 d. At 15 d, all hens were fed a layer diet (17% CP), and production performance was measured for 20 wk. Feed intake was depressed (P < 0.05) in the CASS treatment compared with BROK and CRN treatments during the 14 d. The most significant BW loss occurred in the CASS treatment (21.9%), whereas BROK and CRN treatments were 7.6 and 9.3%, respectively. The CASS treatment resulted in total cessation of egg production within 7 d, whereas those of BROK and CRN treatments decreased to 3.7 and 8.7%, respectively. Ovary weights, oviduct weights, and oviduct lengths of the CASS treatment were not significantly different from those of BROK and CRN treatments (P > 0.05). The weight of spleen and thymus glands, blood heterophil:lymphocyte ratios, bone parameters, and mortality during the 14-d treatment were not affected by treatments. No significant differences were found among treatments for postmolt cumulative egg production, egg weight, feed intake, and mortality. Significant (P < 0.05) improvements in albumen quality were observed in the CASS treatment, although shell thickness of this treatment was reduced (P < 0.05) compared with the CONT treatment in wk 2 after the 14-d treatment. With respect to shell ultrastructural traits, the palisade layer of CASS eggs was shorter (P < 0.05) than that of CONT eggs determined in wk 2 after the 14-d treatment. In addition, evidence of abnormal mammillae was detected in an egg produced by the CASS hens. This research indicates that cassava diets can induce molt in hens. Nonetheless, the technique needs to be adjusted to improve postmolt egg production and shell quality. PMID:24046430

Gongruttananun, N; Guntapa, P; Saengkudrua, K



Protection against Hemorrhagic Colitis in an Animal Model by Oral Immunization with Isogeneic Rabbit Enteropathogenic Escherichia coli Attenuated by Truncating Intimin  

PubMed Central

Strains of Shiga toxin (Stx)-producing Escherichia coli, also called enterohemorrhagic E. coli (EHEC), are important food-borne pathogens for humans. Most EHEC strains intimately adhere to the intestinal mucosa in a characteristic attaching and effacing (A/E) pattern, which is mediated by the bacterial adhesin intimin. Subsequent release of Stx1 and/or Stx2 leads to the frequent development of hemorrhagic colitis and, less commonly, to hemolytic-uremic syndrome. The aim of the present study was to develop an attenuated A/E E. coli strain for use as a vaccine against EHEC infection encoding a truncated intimin lacking adhesive capacity, but which would still express somatic antigens, other products of the locus of enterocyte effacement pathogenicity island, and an immunogenic remnant of the intimin molecule. A single-nucleotide deletion was generated in the eae gene in the prototype rabbit A/E E. coli strain RDEC-1 (O15:H?), which resulted in truncation of intimin by 81 C-terminal residues (860 to 939 amino acids) containing a disulfide loop. Inoculation of rabbits with large doses of the truncated intimin mutant (RDEC-1?eae860-939) was well tolerated, as observed by the absence of clinical signs of disease or evidence of intestinal A/E lesions. The efficacy of RDEC-1?eae860-939 as a vaccine was evaluated by orogastric inoculation of rabbits with RDEC-1?eae860-939 followed by challenge with the virulent strain RDEC-H19A, an Stx1-producing derivative of wild-type RDEC-1 capable of inducing hemorrhagic colitis in rabbits. Following RDEC-H19A challenge, nonimmunized control rabbits exhibited characteristic weight loss with watery to bloody diarrhea and demonstrated intimate bacterial attachment, effacement of microvilli, submucosal edema, mucosal heterophile infiltrates, and Shiga toxin-induced vascular lesions. In contrast, the RDEC-1?eae860-939-immunized rabbits showed no clinical signs of disease, maintained normal weight gain, had reduced fecal shedding of challenge organisms, and showed an absence of gross or microscopic lesions in the intestinal mucosa. Serum antibodies specific to intimin were detected among rabbits immunized with RDEC-1?eae860-939, indicating that truncation of the intimin functional domain not only attenuated bacterial virulence, but also retained at least some of the immunogenicity of native intimin. Although it is not possible to gauge the exact contribution of residual intimin immunity to protection, this attenuation strategy for A/E E. coli strains shows promise for the development of effective vaccines to prevent EHEC infection in humans and animals.

Agin, Tonia S.; Zhu, Chengru; Johnson, Laura A.; Thate, Timothy E.; Yang, Zhuolu; Boedeker, Edgar C.



Time-resolved fluoroimmunoassays of the complete set of secreted phospholipases A2 in human serum.  


Time-resolved fluoroimmunoassays (TR-FIA) were developed for all human secreted phospholipases A(2) (PLA(2)), viz. group (G) IB, GIIA, GIID, GIIE, GIIF, GIII, GV, GX and GXIIA PLA(2) and the GXIIB PLA(2)-like protein. Antibodies were raised in rabbits against recombinant human PLA(2) proteins and used in sandwich-type TR-FIAs as both catching and detecting antibodies, the latter after labeling with Europium. The antibodies were non-cross-reactive. The analytical sensitivities were 1 microg/L for the TR-FIA for GIB PLA(2), 1 microg/L (GIIA), 35 microg/L (GIID), 3 microg/L (GIIE), 4 microg/L (GIIF), 14 microg/L (GIII), 11 microg/L (GV), 2 microg/L (GX), 92 microg/L (GXIIA) and 242 microg/L (GXIIB). All secreted PLA(2)s were assayed by these TR-FIAs in serum samples from 34 patients (23 men and 11 women, mean age 53.2 years) treated in an intensive care unit for septic infections, and in control samples from 28 volunteer blood donors (14 men and 14 women, mean age 57.0 years). Five serum samples (3 in the sepsis group and 2 in the blood donor group) gave high TR-FIA signals that were reduced to background (blank) levels by the addition of non-immune rabbit IgG to the sera. This reactivity was assumed to be due to the presence of heterophilic antibodies in these subjects. In all other subjects, including septic patients and healthy blood donors, the TR-FIA signals for GIID, GIIE, GIIF, GIII, GV, GX and GXIIA PLA(2) and the GXIIB PLA(2)-like protein were at background (blank) levels. Four patients in the sepsis group had pancreatic involvement and elevated concentration of GIB PLA(2) in serum (median 19.0 microg/L, range 13.1-33.7 microg/L, n = 4) as compared to the healthy blood donors (median 1.8 microg/L, range 0.8-3.4 microg/L, n = 28, P < 0.0001). The concentration of GIIA PLA(2) in the sera of septic patients (median 315.7 microg/L, range 15.9-979.6 microg/L, n = 34) was highly elevated as compared to that of the blood donors (median 1.8 microg/L, range 0.8-5.8 microg/L, n = 28, P < 0.0001). Our current results confirmed elevated concentrations of GIB and GIIA PLA(2) in the sera of patients suffering from acute pancreatitis or septic infections, respectively, as compared to healthy subjects. However, in the same serum samples, the concentrations of the other secreted PLA(2)s, viz. GIID, GIIE, GIIF, GIII, GV, GX and GXIIA PLA(2) and the GXIIB PLA(2)-like protein were below the respective analytical sensitivities of the TR-FIAs. It is concluded that generalized bacterial infections do not lead to elevated serum levels of GIIE, GIIF, GIII, GV and GX PLA(2)s above the detection limits of the current TR-FIAs. PMID:15863368

Nevalainen, Timo J; Eerola, Leena I; Rintala, Esa; Laine, V Jukka O; Lambeau, Gérard; Gelb, Michael H