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Background and aims: The aim of this randomized trial in the acute porcine model was to compare the quality of transgastric peritoneoscopy with the use of low-pressure versus standardpressure pneumoperitoneum and to evaluate the respective associated cardiopulmonary changes. Methods: For transgastric peritoneoscopy, carbon dioxide was insufflated via the endoscope for a constant intraperitoneal pressure of 6mmHg or 12mmHg in 9 pigs each. The quality of transgastric peritoneoscopy was rated on a visual analog scale (0 mm, min.; 100 mm, max.) by the endoscopist, who was blinded to the intraperitoneal pressure. The cardiac index and global end-diastolic volume index (GEDVI, reflecting preload) were measured every 3 minutes by transpulmonary thermodilution. The following were also recorded: heart rate, mean arterial pressure (MAP), systemic vascular resistance index (SVRI, reflecting afterload), peak inspiratory pressure (PIP), pH, PCO2, and PO2. Results: The quality of transgastric peritoneoscopy with the use of low-pressure pneumoperitoneum was not inferior to that obtained using standard-pressure pneumoperitoneum (87.0mm vs. 87.3 mm; P < 0.05). In both groups we observed a statistically significant rise in MAP and SVRI. The increase in SVRI was less pronounced during low-pressure peritoneum (P = 0.042), indicating a reduced stress response in comparison to standard-pressure peritoneum. There were no relevant differences between the groups in relation to cardiac index, GEDVI, and heart rate. An intra-abdominal pressure of 6mmHg also led to better oxygenation (P = 0.031 for difference in PO2 between the two groups) due to lower peak inspiratory pressure (P < 0.001 for difference). There were only slight differences between the groups with regard to pH and PCO2. Conclusions: Pneumoperitoneum of 12-16mmHg is used for standard laparoscopy. For NOTES, low-pressure pneumoperitoneum is sufficient and is associated with an improved cardiopulmonary response compared to standard-pressure pneumoperitoneum. [ABSTRACT FROM AUTHOR]

Using ruminally cannulated steers, we investigated how urinary allantoin excretion was related to variations in feed intake and stage of forage maturity. Further, different approaches were compared for predicting ruminal microbial crude protein (MCP) synthesis and its efficiency. Experimental diets were arranged in a replicated 3 × 3 Latin square design (experiment 1) and a 4 × 4 Latin square design (experiment 2). In experiment 1, a mixed diet [forage to concentrate, 68:32 on a dry matter (DM) basis] was fed at three intake levels corresponding to 1, 1.5 and 2 times maintenance energy requirements. In experiment 2, four silage-based diets were fed based on perennial ryegrass ( Lolium perenne L.), which was harvested at four maturity stages. Both experiments demonstrated the influence of diet on microbial growth rate and by this on efficiency of MCP synthesis, although the magnitude of the effects differed between approaches used for estimating MCP. Linear functions satisfactorily characterised the relationship between urinary allantoin excretion ( y) and digestible organic matter (OM) intake ( x, kg/day; experiment 1: y = 7.94 + 17.34 x; R2 = 0.785) or intake of OM effectively degraded in the rumen ( x, kg/day; experiment 2; y = 22.32 + 5.93 x; R2 = 0.695). Urinary excretion of allantoin permitted a semi-quantitative prediction of MCP synthesis: ranking of diets and magnitude of changes in MCP synthesis were reflected. [ABSTRACT FROM AUTHOR]

Fasting dogs do transport vitamin A (VA) in plasma not only as retinol but predominantly as retinyl esters. Contrary to retinol, nothing is known concerning the effects of athletic performance on plasma retinyl ester concentrations. The aim of this study was therefore to examine whether physical stress because of exercise and modification of the oxidative stress by supplementation of α-tocopherol influences the concentrations of retinol and retinyl esters in plasma of sled dogs. The study was carried out on 41 trained adult sled dogs, which were randomly assigned into two groups. One group (19 dogs) was daily substituted with 50 mg dl- α-tocopheryl acetate per kilogram body weight and the control group (22 dogs) was maintained on a basal diet during 3 months prior to exercise. The plasma concentrations of retinol, retinyl esters, α-tocopherol and triglycerides were measured immediately before, directly after and 24 h after exercise. The supplementation of α-tocopheryl acetate had no effect on plasma retinol and retinyl ester concentrations at any measurement time point. However, retinyl ester levels doubled in the non-supplemented group immediately after the race (p < 0.001), whereas in the supplemented group similar high levels were observed not until 24 h post-racing (p < 0.001). The high levels of retinyl esters were paralleled to some extent by an increase in plasma triglyceride concentrations, which were significantly higher 24 h post-racing than immediately before (p < 0.001) and after exercise (p < 0.001) in both groups. The increase in retinyl ester concentrations might be indicative of their mobilization from liver and adipose tissue. Whether plasma retinyl esters can be used as an indicator for the extent of nutrient mobilization during and post-exercise in sled dogs remains to be elucidated. [ABSTRACT FROM AUTHOR]

To measure the immunoglobulin G (IgG) concentration in colostrum, milk and serum samples, a sandwich enzyme-linked immunosorbent assay (ELISA) detection system was developed. The system provided high reproducibility and sensitivity for routine diagnostic purposes. The period of fluctuating serum concentrations of IgG was monitored in new-born foals and their mares for a period of 6 weeks postnatum and postpartum, respectively. All foals received colostrum from their mares. The mean IgG concentration in the precolostral mare serum was approximately 19.0 mg/ml and decreased significantly to 13.8 mg/ml within the first 24 h postpartum. The IgG value fell to a minimum of 11.2 mg/ml by day 21 and increased to 21.6 mg/ml by day 42 postpartum. Within the first 4 h postpartum, mean IgG concentrations of 54.5 mg/ml were measured in the colostrum. A significant decrease to 10.1 mg IgG/ml colostrum was then noted 9–12 h postpartum. The mean IgG concentrations in foal serum increased from 0.3 mg/ml (precolostral value) to 9.6 mg/ml within 5–8 h postnatum. After 13–16 h postnatum, the highest IgG value of 15.7 mg/ml was reached. Over time the mean IgG concentration decreased significantly to 7.9 mg/ml at day 35. At the end of the observation period (day 42 postnatum) the mean IgG concentration once again increased to 11.2 mg/ml serum. In addition, the possible influence of various parameters on IgG concentration were examined. No significant influences could be shown by the breed, mare age, number of pregnancies, days of gestation, month foaled, foal sex, or the different farms. Finally, the cumulative incidence of failure of passive transfer (FPT) defined as IgG levels < 4 mg/ml foal serum, and partial FPT (PFPT) at levels ranging from 4 to 8 mg/ml foal serum was determined. From a total of 70 foals, 10.0% showed FPT and 18.6% showed PFPT. [ABSTRACT FROM AUTHOR]

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Freund's Complete Adjuvant in the Chicken: Efficient Immunostimulation but Severe Local Inflammation To quantify the enhancing activity of Freund's complete adjuvant (FCA), the humoral immune response of hens to the antigen human serum albumin (HSA) injected intramuscularly combined with FCA was measured. Local tissue reaction was examined on macroscopic and microscopic level. Other hens were injected with HSA in physiological sodium-chloride solution and served as controls. The immunization of hens without using FCA resulted in a short-term rise in immunoglobulin G (IgG) antibody titres, while the hens injected with HSA and FCA developed a long-term response (plateau-form of antibody titres). Local tissue reaction after injection of HSA in sodium-chloride solution was a mild and transient form of interstitial myositis. The immunization with HSA combined with FCA regularly resulted in a persisting granulomatous myositis. It is therefore a matter of animal protection to demand restricted use of FCA and to promote adjuvants of comparable efficiency, but better tolerance. Zusammenfassung In der vorliegenden Arbeit wurden die humorale Immunantwort von Hühnern gegenüber dem intramuskulär in Kombination mit Freundschem komplettem Adjuvans (FCA) verabreichten Antigen humanes Serumalbumin (HSA) quantifiziert und lokale Gewebereaktionen auf makroskopischer und lichtmikroskopischer Ebene untersucht. Zum Vergleich wurde HSA in physiologischer Kochsalzlösung appliziert. Die Immunisierung von Hühnern ohne FCA führte zu zeitlich sehr begrenzten IgG-Antikörpertitern. Die Verwendung von FCA bei der Immunisierung mit HSA resultierte in der Ausbildung eines langanhaltenden, hohen Antikörpertiter-Plateaus. Die lokale Gewebereaktion nach Immunisierung mit HSA in physiologischer Kochsalzlösung war durch eine vorübergehende geringgradige interstitielle Myositis gekennzeichnet. Die Immunisierung unter Verwendung von FCA hatte regelmäßig die Entwicklung einer persistierenden granulomatösen Myositis zur Folge. Uinter dem Aspekt des Tierschutzes ist deshalb zu fordern, auf die Verwendung von FCA soweit als möglich zu verzichten und stattdessen besser verträgliche aber vergleichbar effiziente Adjuvantien zu entwickeln und einzusetzen. [ABSTRACT FROM AUTHOR]

The Influence of Rumen Metabolism on Cyclic N-Compounds in the Blood Plasma of Ruminants The purpose of this paper was to study the effect of rumen metabolism on the concentration of certain cyclic N-compounds in blood plasma. The plasma from two groups of lambs in the phase of rumen development during early and late weaning was used. As a second model, samples from steers fed at maintenance and after emptying of the rumen via cannula were used. Plasma samples were obtained from the jugular vein in regular intervals. The HPLC-technique was employed as analytical method. The plasma levels of allantoin and of hippuric acid increased about 2.2-3.7 fold and 2.4-6.4 fold in lambs during rumen development. They decreased to about 43 % and 13 % after emptying the rumen of steers. These metabolites appear to reflect rumen protein synthesis and fibre breakdown and are considered useful estimators of rumen function. Cytidine, uridine, uric acid and hypoxanthine were only influenced in part by rumen metabolism. Plasma pseudouridine was decreased and creatinine was increased in lambs in relation to body mass and remained unaffected in steers. Zusammenfassung Um den Einfluß des Pansenstoffwechsels auf bestimmte zyklische N-Verbindungen im Blutplasma von Wiederkäuern zu klären, wurden Proben aus zwei Untersuchungsreihen gewonnen und mittels HPLC analysiert: erstens von Lämmern vor und nach früher bzw. später Entwöhnung; zweitens von Ochsen mit Pansenkanülen vor und nach Entleerung des Pansens. Die Plasmawerte von Allantoin und Hippursäure erhöhten sich während der Pansenentwicklung 2, 2 bzw. 2, 4 fach (frühentwöhnte Gruppe) und 3, 7 bzw. 6, 4 fach (spätentwöhnte Gruppe). Die Metabolitkonzentration im Plasma von Ochsen nahmen nach Entleerung des Pansens auf rund 43 % (Allantoin) und 13 % (Hippursäure) der Normalwerte ab. Diese Metaboliten können zur Bewertung der Proteinsynthese bzw. der Faserverdauung im Pansen herangezogen werden. Die Konzentrationen von Uridin, Cytidin, Harnsäure und Hypoxanthin im Plasma hängen weniger deutlich vom Pansenstoffwechsel ab und unterliegen auch anderen Einflüssen. Pseudouridin und Kreatinin im Plasma waren unabhängig vom Pansenstoffwechsel. Ihre Konzentrationen fielen (Pseudouridin) bzw. stiegen (Kreatinin) mit der Körpermasse. [ABSTRACT FROM AUTHOR]

On the action of a glucocorticoid on glucose metabolism of pygmy goats 1st Communication: Some metabolites of energy metabolism In order to study mechanisms in ruminants responsible for glucocorticoid-induced hyperglycemia the effects of a single intravenous dose of a glucocorticoid on energy metabolism were investigated by measuring plasma levels of glucose, lactate, free fatty acids (FFA), 3-hydroxybutyrate (3-HOB) and acetoacetate (AcAc). Experiments were performed on four castrated male African pygmy goats at two metabolic states: 'fed' with a single daily meal of hey and 'fasted' with starvation for 3 days. The untreated goats served as their own control. The glucocorticoid application to fed animals resulted in an increase in glucose level (+43%) lasting more than 11 h and an irregular increase of lactate. The concentration of 3-HOB was increased especially before feeding (+64%). In contrast, FFA were decreased (-27%) the second day after glucocorticoid application. In fasted goats the glucocorticoid also caused an increase in glucose level (+ 25%). The effect was smaller than in fed animals. Concentrations of FFA, 3-HOB and AcAc were 10, 20 and 35 % lower than those of untreated goats. With regard to the antiketotic use of glucocorticoids the decrease of plasma levels of FFA and ketone bodies in starving animals is of special importance. Obviously the animals were able to use glucose again as a main energy source instead of FFA. Zusammenfassung Um bei Wiederkäuern Einblick in die für die Glucocorticoid-induzierte Hyperglykämie verant-wortlichen Mechanismen zu gewinnen, wurden die Auswirkungen einer einmaligen Glucocorticoid-Behandlung auf den allgemeinen Energiestoffwechsel anhand von Blutspiegelmessungen der Metabolite Glucose, Lactat, freie Fettsäuren (FFS), 3-Hydroxybutyrat (3-HOB) und Acetacetat (AcAc) untersucht. Als Versuchstiere wurden vier kastrierte Böcke der Rasse 'Afrikanische Zwergziege' zum einen im Stoffwechselzustand 'gefüttert' mit einer einmal täglichen Heu-Fütterung und zum anderen nach 3tägigem Futterentzug im Stoffwechselzustand 'hungernd' entweder unbehandelt oder nach intravenöser Glucocorticoid-Verabreichung untersucht. Die Glucocorticoid-Behandlung verursachte bei den gefütterten Tieren einen plateauartigen Anstieg (43%) des Glucosespiegels und einen unruhigen, insgesamt ansteigenden Verlauf der Lactat-konzentration. Die 3-HOB-Werte waren insbesondere vor der Futteraufnahme um 64% erhöht. Demgegeniiber zeigten die FFS erst am 2. Tag nach der Behandlung einen um 27% erniedrigten Tagesspiegel. Bei hungernden Tieren verursachte die Glucocorticoid-Behandlung ebenfalls einen Anstieg des Glucosespiegels (25%), der allerdings geringer ausfiel als bei gefütterten. Die Konzentration der FFS lag um 10%, die von 3-HOB um 20% und die von AcAc um 35% unter den Werten der unbehandelten Gruppe. Im Hinblick auf den antiketogenen Einsatz von Glucocorticoiden kommt dem Absinken der Plasmaspiegel der FFS und der Ketonkörper bei hungernden Tieren besondere Bedeutung zu. Den Tieren war es mit Einsetzen der Hyperglykämie offensichtlich möglich, ihren Energiebedarf wieder vermehrt über Glucose zu decken und die Verstoffwechslung alternativer Energiesubstrate (FFS) zu reduzieren. [ABSTRACT FROM AUTHOR]

Effects of a Glucocorticoid on Glucose Metabolism in Pygmy Goats 2. Glucose Turnover and Recycling In the present study glucose kinetic parameters like pool size, distribution volume and rates of total entry and irreversible loss were determined in four castrated male African pygmy goats during hyperglycemia induced by an intravenous glucocorticoid injection (0. 25 mg dexamethasone = DXM per kg body weight). Experiments were done at two different metabolic states, 'fed' once daily and 'fasted' after three days of starvation. The use of [U-14C]- together with [2-3H]- or [6-3H]-glucose made it possible to show changes in rates of gluconeogenesis, phosphorylation and specific recycling of glucose-C (e. g. lactate/alanine). During glucocorticoid induced hyperglycemia (+ 45%) in fed animals de novo synthesis of glucose (+5%) and lactate/alanine recycling increased (+4%). Rates of irreversible loss for all applied tracers in fasted animals formed only two thirds of the values of fed animals. During hyperglycemia (+40%) after DXM-injection de novo synthesis of glucose (+8%) and lactate/alanine recycling also increased (+27%). Furthermore the rate of phosphorylation of glucose decreased in fasted pygmy goats. The increase in availability of glucose both in fed and fasted animals is due to an increase in gluconeogenesis. In fasted animals additionally a decrease in the rate of phosphorylation and therefore of the peripheral glucose turnover has to be considered. Zusammenfassung In der vorliegenden Untersuchung wurden Pool, Verteilungsraum, Gesamteintrittsrate (GER) und irreversibler Verlust (IRL) als Parameter des Glucosestoffwechsels an vier kastrierten männlichen Afrikanischen Zwergziegen bei einer durch Glucocorticoid (0,25 mg Dexamethason / kg KGW, intravenös) induzierten Hyperglykämie bestimmt. Die Versuche wurden bei einmal täglich gefütterten Tieren und nach 3tägigem Futterentzug (hungernd) durchgeführt. Der Einsatz von [U-14C]- und [2-3H]- oder [6-3H]-Glucose ermöglichte die zusätzliche Darstellung von Veränderungen bei der Gluconeogenese, der Phosphorylierungsrate und spezifischer Recyclingraten von Glucose (z. B. über Lactat/Alanin). Bei den gefütterten Tieren war wahrend der Glucocorticoid-induzierten Hyperglykämie (+45%) die Neusyntheserate von Glucose um 5% und das Lactat/Alanin-Recycling um 4% erhöht. Die irreversiblen Verluste der hungernden Tiere lagen fur alle venvendeten Glucosemar-kierungen um ein Drittel niedriger als die Vergleichswerte gefiitterter Tiere. Nach DXM-Behandlung stieg wahrend der Hyperglykamie (+40%) auch die Neusynthese von Glucose um 8% und das Lactat/Alanin-Recycling um 27% an. Zusätzlich nahm in diesem Stoffwechselzustand die Phosphorylierungsrate von Glucose um 20% ab. Die bei beiden Stoffwechselzustanden festgestellte Zunahme der Glucoseverfugbarkeit ist auf eine Steigerung der Gluconeogenese zurückzuführen, wobei im Hungerzustand zusätzlich eine Abnahme der Phosphorylierungsrate und damit des peripheren Glucoseumsatzes zu berücksichtigen ist. [ABSTRACT FROM AUTHOR]

Focuses on the absorption and synthesis of immunoglobulins G in newborn calves in Germany. Importance of colostrum for the reduction of newborn mortality; Consideration of the body weight gain of the calves; Suggestion of further investigations on immunoglobulin absorption.

Editorial. Pays tribute to Manfred Kirchgebner who will resign from active duty as chief editor of the 'Journal of Animal Physiology and Animal Nutrition,' and become honorary editor of the journal at the end of December 2002. Career achievements; Dedication to the journal; Commitment to improving the content of the journal.

The time course of serum concentrations of the bovine IgG1 and IgG2 was monitored in 18 newborn colostrum-fed calves from birth to 11 weeks of age. All calves received three 1.5 l meals of a pooled colostrum with IgGl and IgG2 concentrations of 54.9 mg/ml and 4.2 mg/ml during the first 14 h post-natum. The mean IgG concentrations in calf serum increased from 0.15 mg IgG1/ml and 0.06 mg IgG2/ml (precolostral values) to 9.3 mg IgG1/ml and 0.8 mg IgG2/ml 12 h after the third colostrum meal. Thereafter, IgG1 decreased continuously to a minimum level of 4.9 mg/ml (p < 0.05) at day 28 post-natum and increased to 9.0 mg/ml at day 77. Postcolostral mean IgG2 was lowest (0.5 mg/ml) at day 11 and highest (1.2 mg/ml) at day 77. With these postcolostral IgG concentrations the respective body weight-corrected serum IgG pools approximately were calculated. According to that procedure the initially high IgG pool in the serum at day 2 corresponds to 11.3 and 8.7%, respectively, of the ingested colostral IgG1 and IgG2. The time course of these IgG pools in the serum could be characterized by four typical phases. After a rapid increase from 0.45 g to 22.6 g (12 h after the last colostrum meal) in phase I, the total serum IgG decreased to 17.6 g at day 11 post-natum (phase II) and levelled-out until day 28 post-natum at a value of 17.3 g (phase III). In phase IV total serum IgG increased to 49.3 g at the end of the observation period (week 11). Considering these IgG values, which had been standardized by body weight, the endogeneous IgG production seemed to start clearly (at week 1–2 post-natum) before the increase of the IgG concentration in the serum. To measure these IgG1 and IgG2 concentrations in the serum as well as in the colostrum, sandwich ELISA detection systems were developed which specifically quantify these IgG subclasses in cattle. The specific immunoglobulin Y (IgY) could be isolated from the egg yolk of laying hens which have been immunized with bovine IgG1... [ABSTRACT FROM AUTHOR]

Large animal ischemic cardiomyopathy models are widely used for preclinical testing of promising novel therapeutic approaches. Pressure volume (PV) loop analysis and cardiac magnetic resonance imaging (CMRI) allow functional and morphological phenotyping. In this study we performed a comparative analysis of both methods highlighting the strength of each and their synergistic potential. Myocardial infarction (MI) was created in German farm pigs (German Landrace) by 2 h LCX occlusion (n = 11) and subsequent reperfusion. Cardiac function was assessed by PV-loops and CMRI 56 and 112 days post-MI. Two hours occlusion of the LCX led to mid-size left ventricular (LV) MI represented by high-sensitive troponin T (hsTnT) 3 days post-MI, correlating well with cardiac CMRI late enhancement. CMRI determined end-diastolic and end-systolic volumes significantly increased post-MI, while ejection fraction was reduced in infarcted animals compared to the sham group (n = 6). PV-loop derived preload-insensitive parameters of systolic and diastolic function were diminished post-MI compared to sham animals while preload-dependent parameters only deteriorated in advanced HF. PV-loop analysis significantly correlates with CMRI analysis of cardiac function in pig post-MI ischemic cardiomyopathy. PV-Loop analysis accurately quantifies LV volumetry and function in post-MI HF, and thus eccentric LV morphology. PV-loop analysis correlates well to cardiac MRI. Preload-insensitive parameters show high sensitivity to quantify HF while preload-sensitive parameters are not able to quantify early-stages of LV HF.

Depletion of B cells with the anti-CD20 antibody rituximab is an established therapy for rheumatoid arthritis. However, rituximab has only moderate efficacy, most likely due to insufficient depletion of B cells in lymphoid organs and expansion of pathogenic B cells. We found that an antibody against mouse CD79b profoundly blocks B-cell proliferation induced via the B-cell receptor, CD40, CD180, and chondroitin sulfate, but not via TLR4 or TLR9. Treatment with anti-CD79b also induces death in resting and activated B cells. B-cell inhibition is mediated by cross-linkage of CD79b, but independent of Fc-receptor engagement. In the model of collagen-induced arthritis, an antibody against mouse CD20 depletes B cells very efficiently but fails to suppress the humoral immune response against collagen and the development of arthritis. In contrast, the antibody against CD79b, and a deglycosylated variant of this antibody, almost completely inhibits the increase in anti-collagen antibodies and the development of arthritis. In mice with established arthritis only the fully glycosylated antibody against CD79b is effective. Our data show that targeting B cells via CD79b is much more effective than B-cell depletion with anti-CD20 antibodies for therapy of arthritis. These findings may have important implications for treatment of B-cell-mediated autoimmune diseases., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Human umbilical vessels have been recognized as a valuable and widely available resource for vascular tissue engineering. Whereas endothelium-denuded human umbilical veins (HUVs) have been successfully seeded with a patient-derived neoendothelium, decellularized vessels may have additional advantages, due to their lower antigenicity. The present study investigated the effects of three different decellularization procedures on the histological, mechanical and seeding properties of HUVs. Vessels were decellularized by detergent treatment (Triton X-100, sodium deoxycholate, IGEPAL-CA630), osmotic lysis (3 m NaCl, distilled water) and peroxyacetic acid treatment. In all cases, nuclease treatments were required to remove residual nucleic acids. Decellularization resulted in a partial loss of fibronectin and laminin staining in the subendothelial layer and affected the appearance of elastic fibres. In addition to removing residual nucleic acids, nuclease treatment weakened all stainings and substantially altered surface properties, as seen in scanning electron micrographs, indicating additional non-specific effects. Detergent treatment and osmotic lysis caused failure stresses to decrease significantly. Although conditioned medium prepared from decellularized HUV did not severely affect endothelial cell growth, cells seeded on decellularized HUV did not remain viable. This may be attributed to the partial removal of essential extracellular matrix components as well as to changes of surface properties. Therefore, decellularized HUVs appear to require additional modifications in order to support successful cell seeding. Replacing the vessels' endothelium may thus be a superior alternative to decellularization when creating tissue-engineered blood vessels with non-immunogenic luminal interfaces., (Copyright © 2012 John Wiley & Sons, Ltd.)

We are today surrounded almost constantly by high-frequency electromagnetic fields (EMFs) from mobile communications base stations. To date, however, there has been little concern regarding nonthermal effects of EMFs on cognition. In the present study, male and female rats were subjected to continuous far-field exposure to a frequency of 900-MHz (Global System for Mobile Communications [GSM]) or 1.966-GHz (Universal Mobile Telecommunications System [UMTS]) at 0.4 W/kg. Memory performance of adult EMF-exposed and sham-exposed female rats (at 6 months of age) and male rats (at 3 and 6 months of age) was tested using a social discrimination procedure. For this procedure, a target juvenile male was introduced to the subject's home cage for 4 min (Trial 1). After 30 min, the same target animal and a novel juvenile male were simultaneously presented to the subject for 4 min (Trial 2). Differences in sniffing duration to the familiar and novel target rats during Trial 2 were used to assess memory performance. EMF-exposed females exhibited no differences in sniffing duration compared with controls. In contrast, the sniffing durations of EMF-exposed males at 3 months of age were significantly affected. At 6 months of age, GSM-, but not UMTS-, exposed male adults showed a memory performance deficit. These findings provide new insight into the nonthermal effects of long-term high-frequency EMF exposure on memory., (PsycINFO Database Record (c) 2014 APA, all rights reserved.)

Glycosaminoglycans have anti-inflammatory properties and interact with a variety of soluble and membrane-bound molecules. Little is known about their effects on B cells and humoral immune responses. We show that CS but not dextran or other glycosaminoglycans induces a pronounced proliferation of B cells in vitro compared with TLR4 or TLR9 ligands. With the use of inhibitors and KO mice, we demonstrate that this proliferation is mediated by the tyrosine kinases BTK and Syk but independent of CD44. Antibodies against Ig-α or Ig-β completely block CS-induced B cell proliferation. Injection of CS in mice for 4-5 days expands B cells in the spleen and results in a marked increase of CD138(+) cells in the spleen that is dependent on BTK but independent of CD4(+) T cells. Long-term treatment with CS for 14 days also increases CD138(+) cells in the bone marrow. When mice were immunized with APC or collagen and treated with CS for up to 14 days during primary or after secondary immune responses, antigen-specific humoral immune responses and antigen-specific CD138(+) plasma cells in the bone marrow were reduced significantly. These data show that CD138(+) cells, induced by treatment with CS, migrate into the bone marrow and may displace other antigen-specific plasma cells. Overall, CS is able to interfere markedly with primary and fully established humoral immune responses in mice., (© 2014 Society for Leukocyte Biology.)

Autoimmune uveitis is characterized by crossing of blood-retinal barrier (BRB) by autoaggressive immune cells. Equine recurrent uveitis (ERU) is a valuable spontaneous model for autoimmune uveitis and analyses of differentially expressed proteins in ERU unraveled changed protein clusters in target tissues and immune system. Healthy eyes are devoid of leukocytes. In ERU, however, leukocytes enter the inner eye and subsequently destroy it. Molecular mechanisms enabling cell migration through BRB still remain elusive. Previously, we detected decreased talin 1 expression in blood-derived granulocytes of ERU cases, linking the innate immune system to ERU. Because changes in leukocyte protein expression pattern may play a role in pathological abnormalities leading to migration ability, we aimed at identifying interactors of talin 1 in leukocytes with immunoprecipitation, followed by LC-MS/MS for candidate identification. This enabled us to identify CD90 (Thy1) as novel interactor of talin 1 besides several other interactors. In blood-derived granulocytes from healthy individuals, CD90 was highly abundant and significantly reduced in ERU, especially in effector cells. Connection between talin 1 and CD90 and their expression differences in inflammation is an interesting novel finding allowing deeper insight into immune response of innate immune system and granulocyte migration ability in this organ-specific autoimmune disease.

Objective: In equine recurrent uveitis (ERU), immune reactions are directed toward known antigens like S-antigen, interphotoreceptor retinoid-binding protein, and cellular retinalaldehyde-binding protein, and anti-retinal antibodies were detected in vitreous samples. The aim of this study was the investigation of intraocular immunoglobulin M (IgM) reactivities to retinal proteome., Procedures: Retina was separated by one- and two-dimensional gel electrophoresis and blotted semidry on PVDF membranes. To identify intraocular IgM antibody responses to retinal tissue, blots were incubated with vitreous samples of ERU-diseased horses (n = 50) and healthy controls (n = 30), followed by an HRP-labeled secondary antibody specific for equine IgM. Noticeable 2D western blot signals were aligned on a 2D gel of retinal proteome, excised, and subsequently identified by tandem mass spectrometry., Results: Interestingly, frequent and very miscellaneous IgM response patterns to the retinal proteome in 68% of ERU vitreous samples were detected. Binding of IgM antibodies was localized at 17 different molecular weights. The most frequently detected signal, in 21 of the 50 samples, was located at 49 kDa. Comparing the samples interindividually between one and up to nine different signals in one sample could be observed. All healthy vitreous samples were devoid of IgM antibodies. Analysis of targeted spots with mass spectrometry led to the clear identification of 11 different proteins (corresponding to 16 different spots). One candidate could not be discovered so far., Conclusion: The considerable IgM response to retinal proteins demonstrates an ongoing immune response, which might contribute to the remitting relapsing character of ERU. Novel identified target proteins point to a diverse response pattern of individual ERU cases., (© 2012 American College of Veterinary Ophthalmologists.)

The endothelial barrier controls the passage of solutes from the vascular space. This is achieved through active reorganization of the actin cytoskeleton. A central cytoskeletal protein involved into this is vasodilator-stimulated phosphoprotein (VASP). However, the functional role of endothelial VASP during hypoxia has not been thoroughly elucidated. We determined endothelial VASP expression through real-time PCR (Rt-PCR), immunhistochemistry, and Western blot analysis during hypoxia. VASP promoter studies were performed using a PGL3 firefly luciferase containing plasmid. Following approval by the local authorities, VASP ( -/- ) mice and littermate controls were subjected to normobaric hypoxia (8% O(2), 92% N(2)) after intravenous injection of Evans blue dye. In in vitro studies, we found significant VASP repression in human microvascular and human umbilical vein endothelial cells through Rt-PCR, immunhistochemistry, and Western blot analysis. The VASP promoter construct demonstrated significant repression in response to hypoxia, which was abolished when the binding of hypoxia-inducible factor 1 alpha was excluded. Exposure of wild-type (WT) and VASP ( -/- ) animals to normobaric hypoxia for 4 h resulted in an increase in Evans blue tissue extravasation that was significantly increased in VASP ( -/- ) animals compared to WT controls. In summary, we demonstrate here that endothelial VASP holds significant importance for endothelial barrier properties during hypoxia.

Equine recurrent uveitis serves as a spontaneous model for human autoimmune uveitis. Unpredictable relapses and ongoing inflammation in the eyes of diseased horses as well as in humans lead to destruction of the retina and finally result in blindness. However, the molecular mechanisms leading to inflammation and retinal degeneration are not well understood. An initial screening for differentially regulated proteins in sera of uveitic cases compared to healthy controls revealed an increase of the alternative pathway complement component factor B in ERU cases. To determine the activation status of the complement system, sera were subsequently examined for complement split products. We could demonstrate a significant higher concentration of the activation products B/Ba, B/Bb, Bb neoantigen, iC3b and C3d in uveitic condition compared to healthy controls, whereas for C5b-9 no differences were detected. Additionally, we investigated complement activation directly in the retina by immunohistochemistry, since it is the main target organ of this autoimmune disease. Interestingly, infiltrating cells co-expressed activated factor Bb neoantigen, complement split product C3d as well as CD68, a macrophage marker. In this study, we could demonstrate activation of the complement system both systemically as well as in the eye, the target organ of spontaneous recurrent uveitis. Based on these novel findings, we postulate a novel role for macrophages in connection with complement synthesis at the site of inflammation., (Copyright © 2010 Elsevier GmbH. All rights reserved.)

Purpose: Equine recurrent uveitis (ERU) is an incurable disease affecting the inner eye that leads to blindness, through activated T cells that pass the blood-retinal barrier and destroy the retina. Serum markers are a desirable choice for monitoring development of disease, as serum is easy accessible and the markers could serve to predict the beginning of disease or an imminent relapse., Methods: In this study, serum proteomes (depleted of high-abundance serum proteins) of horses with ERU and healthy controls were compared with the 2-D DIGE (two-dimensional gel electrophoresis) technique to identify differentially expressed proteins. The expression pattern of a candidate protein in retina and vitreous was validated by Western blots and immunohistochemistry., Results: Ten differentially expressed proteins could be identified by mass spectrometry (MALDI-TOF/TOF). Five proteins--IgM, IgG4 hc, serotransferrin, alpha-2HS-glycoprotein, and complement factor B--were upregulated in the uveitic state, whereas the five proteins albumin, apolipoprotein A-IV and H, IgG5 hc, and high-molecular-weight kininogen (HK) showed a significantly lower expression in sera of uveitis cases. Of interest, kininogen was significantly upregulated in the target tissues vitreous and retina. HK is a plasma protein with multiple physiological functions, with an important role in inflammation and promoting neovascularization. Most interesting is the as of yet unaddressed association of HK with uveitis. Immunohistochemistry showed coexpression of kininogen and VEGF in inflamed eyes., Conclusions: Since neovascularization plays a major role in the pathogenesis of uveitis, the identification of a proangiogenic factor in the retina presents an important finding and may contribute to elucidating the pathogenesis of uveitis.

Objective: Activation of basophils contributes to memory immune responses and results in exacerbation of collagen-induced arthritis (CIA). We undertook the present study to analyze the production and biologic effects of interleukin-3 (IL-3), a strong activator of basophils, in CIA., Methods: Arthritis was induced by immunization with type II collagen. Mice were treated with blocking monoclonal antibodies against IL-3 or with recombinant IL-3. Clinical scoring, histologic analysis, fluorescence-activated cell sorter analysis, enzyme-linked immunosorbent assay, and cell culturing were performed to assess disease activity and IL-3 production., Results: IL-3 was produced in large quantities by collagen-specific CD4+ T cells in the spleen and was present in the synovial tissue during onset of arthritis, but was down-regulated in paws with severe inflammation. Blockade of IL-3 during the time of arthritis onset resulted in profound improvement of the disease, with reductions in synovial leukocyte and cytokine levels, peripheral blood basophil levels, and anticollagen antibody titers. Blockade of IL-3 during the late phase of arthritis had no beneficial effect. Administration of recombinant IL-3 during onset of arthritis induced a marked exacerbation of the disease, with increased peripheral blood basophil and plasma IL-6 levels and increased titers of anticollagen antibody. In studies of the regulation of IL-3 expression in CD4+ T cells, IL-6 and IL-4 suppressed the release of IL-3 by activated CD4+ T cells, whereas lipopolysaccharide and CpG DNA up-regulated IL-3 secretion in activated CD4+ T cells by acting on costimulatory cells., Conclusion: Taken together, the present results demonstrate for the first time that IL-3 has an important role in the early phase of CIA.