Trans-endocytosis of CD80 and CD86: a molecular basis for the cell-extrinsic function of CTLA-4

Cytotoxic T lymphocyte antigen 4 (CTLA-4) is an essential negative regulator of T cell immune responses whose mechanism of action is the subject of debate. CTLA-4 shares two ligands (CD80 and CD86) with a stimulatory receptor, CD28. Here, we show that CTLA-4 can capture its ligands from opposing cells by a process of trans-endocytosis. After removal, these costimulatory ligands are degraded inside CTLA-4-expressing cells, resulting in impaired costimulation via CD28. Acquisition of CD86 from antigen-presenting cells is stimulated by T cell receptor engagement and observed in vitro and in vivo. These data reveal a mechanism of immune regulation in which CTLA-4 acts as an effector molecule to inhibit CD28 costimulation by the cell-extrinsic depletion of ligands, accounting for many of the known features of the CD28-CTLA-4 system.     

Birds in Western Australian wheatbelt reserves—implications for conservation

Bird species richness in 22 reserves in the Western Australian wheatbelt was shown to be related not to isolation from adjacent uncleared land, either spatially or with time since clearing of land in their vicinity, but to area of reserve and certain reserve habitat variables. The nature of these relationships was examined with multiple regression analysis. Reserve area was the most important variable, except in some passerine groups, where numbers of plant species, vegetation associations, and plant life form and density classes in each vegetation stratum were, separately, more important than area. Eighty-two percent of the variation in the number of bird species was explained by area of reserve and number of plant species, indicating the importance of floristics to the total bird assemblage within reserves. The number of formations present (the broadest vegetation structural grouping used) did not explain any of the variation in species numbers in any of the bird groupings.There were more resident passerine species present in wheatbelt reserves than there were breeding passerine species on similar-sized southwestern Australian late-Pleistocene/Holocene islands, although the species versus area relationship of the resident habitat-specific passerines (P₅ group) was very similar to that of the island land-bird faunas. The P₅ species group contained many species which probably responded to wheatbelt reserves as islands; some of these species may fail to persist in the wheatbelt in the long term. Twenty-three species usually resident in reserves and 15 ‘non-residents’ were identified as being ‘vulnerable’ or of uncertain conservation status in the wheatbelt.Some vegetation formations within reserves were more important to birds than others; woodlands were most important both to resident and transient species. Most species do not appear to distinguish between shrublands and heaths as major habitats; Species richness versus formation area relationship suggest that insular patchiness for birds in the wheatbelt was manifest at the level of vegetation formations. Reserves as small as 80 ha were important sanctuaries for birds in the wheatbelt, although 1500 ha was considered a minimum area of reserve to conserve a local avifauna. Reserves of the order of 30,000–94,000 ha were required to contain most of the avifauna of the wheatbelt.     

Cross-linking and rheological changes of whey proteins treated with microbial transglutaminase

Modification of the functionality of whey proteins using microbial transglutaminase (TGase) has been the subject of recent studies. However, changes in rheological properties of whey proteins as affected by extensive cross-linking with TGase are not well studied. The factors affecting cross-linking of whey protein isolate (WPI) using both soluble and immobilized TGase were examined, and the rheological properties of the modified proteins were characterized. The enzyme was immobilized on aminopropyl glass beads (CPG-3000) by selective adsorption of the biotinylated enzyme on avidin that had been previously immobilized. WPI (4 and 8% w/w) in deionized water, pH 7.5, containing 10 mM dithiothreitol was cross-linked using enzyme/substrate ratios of 0.12-10 units of activity/g WPI. The reaction was carried out in a jacketed bioreactor for 8 h at 40 degrees C with continuous circulation. The gel point temperature of WPI solutions treated with 0.12 unit of immobilized TGase/g was slightly decreased, but the gel strength was unaffected. However, increasing the enzyme/substrate ratio resulted in extensive cross-linking of WPI that was manifested by increases in apparent viscosity and changes in the gelation properties. For example, using 10 units of soluble TGase/g resulted in extensive cross-linking of alpha-lactalbumin and beta-lactoglobulin in WPI, as evidenced by SDS-PAGE and Western blotting results. Interestingly, the gelling point of WPI solutions increased from 68 to 94 degrees C after a 4-h reaction, and the gel strength was drastically decreased (lower storage modulus, G'). Thus, extensive intra- and interchain cross-linking probably caused formation of polymers that were too large for effective network development. These results suggest that a process could be developed to produce heat-stable whey proteins for various food applications.     

Mechanisms of Gastrointestinal Ischemia-Reperfusion Injury and Potential Therapeutic Interventions: A Review and Its Implications in the Horse

Restoration of blood flow after a period of intestinal ischemia is necessary to maintain cell function and viability; however, the reintroduction of oxygen can initiate a cascade of events that exacerbates tissue injury. Intestinal I-R injury is manifested as increased microvascular and mucosal permeability, and mucosal necrosis. Reperfusion injury begins with the accumulation of hypoxanthine from ATP metabolism and the conversion of XDH to XO during ischemia. Upon reperfusion, the XO catalyzes the conversion of hypoxanthine to superoxide radicals in the presence of oxygen. Superoxide radicals are further reduced to highly reactive hydroxyl radicals, which initiate lipid peroxidation. Lipoperoxidation causes functional and structural alterations in cell membrane lipids and can release numerous inflammatory mediators, which exacerbate tissue damage. Neutrophils are recruited into tissues during ischemia and on reperfusion; then they undergo degranulation and release destructive products (proteases and OFRs), which mediate further tissue injury. A limited number of experimental studies in the gastrointestinal tract of horses have shown I-R injury. Additional studies are necessary to further elucidate and sequence the precise pathophysiologic mechanisms occuring in the equine intestine during I-R. Therapy should be focused on prevention of I-R injury by pharmacologic or chemical inhibition or modification of these pathophysiologic pathways. Selected pharmacologic agents or drug combinations may offer novel, scientifically relevant and yet practical approaches to alleviating intestinal I-R injury in horses. This may improve survival of horses with naturally acquired intestinal strangulation obstruction.     

Effect of sodium bicarbonate infusion on serum osmolality, electrolyte concentrations, and blood gas tensions in cats.

The effects of single IV injections of sodium bicarbonate (0.5 mEq/kg of body weight, 1 mEq/kg, 2 mEq/kg, and 4 mEq/kg) on serum osmolality, serum sodium, chloride, and potassium concentrations, and venous blood gas tensions in 6 healthy cats were monitored for 180 minutes. Serum osmolality increased and remained significantly (P less than 0.05) increased for 120 minutes in cats given 4 mEq of sodium bicarbonate/kg. Serum sodium was increased significantly (P less than 0.05) for 30 minutes in cats given 4 mEq of sodium bicarbonate/kg. Serum sodium decreased and remained significantly (P less than 0.05) decreased for 120 minutes in cats given 1 g of 20% mannitol/kg, and serum osmolality was significantly (P less than 0.05) decreased at 30 and 60 minutes. Serum chloride decreased significantly (P less than 0.05) for 10 minutes in cats given 1 mEq of sodium bicarbonate/kg, and was significantly decreased for 30 minutes in cats given 2 mEq and 4 mEq of sodium bicarbonate/kg. Serum chloride decreased and remained significantly (P less than 0.05) decreased for 30 minutes in cats given 1 g of 20% mannitol/kg. Serum sodium and serum osmolality did not change significantly (P less than 0.05) in cats given 4 ml of 0.9% sodium chloride/kg. Serum potassium decreased significantly (P less than 0.05) for 10 minutes in cats given 1 mEq of sodium bicarbonate/kg, and for 120 minutes in cats given 2 mEq/kg or 4 mEq/kg. There was a significantly (P less than 0.05) greater decrease in serum potassium that lasted for 30 minutes after given sodium bicarbonate at the dosage of 4 mEq/kg, compared with other dosages given.(ABSTRACT TRUNCATED AT 250 WORDS)     

A new analgesic drug combination in the horse

A xylazine and butorphanol drug combination produced minimal and transient hemodynamic effects and no significant respiratory depression when administered to 6 healthy horses. Combining xylazine and butorphanol produced a synergistic analgesic effect and provided good chemical restraint for a standing surgical procedure.     

Clinical and laboratory findings in cats infected with feline immunodeficiency virus

Thirty-two cats referred to the Feline Studies Centre between June 1987 and October 1988, and 14 in-contact cats, were found to be infected with feline immunodeficiency virus. Most of the 46 cats were non-pedigree and free ranging; 27 were male (19 neutered) and 19 were female (18 neutered). Their ages ranged from one to 17 years and the average age was 5.8 years. The most common clinical signs were lethargy, inappetence, weight loss, pyrexia and lymphadenopathy; most cases had multiple abnormalities. Other common signs were gingivitis, diarrhoea, rhinitis and ocular discharge. Eight cats had neoplasia. The commonest haematological abnormalities were anaemia, neutropenia, lymphopenia and monocytosis. Eight cats had lymphocytosis; seven of these were in a single house-hold. Several cats had high serum globulin levels and half of those tested had high IgG levels. Seven cats had no detectable antibody to feline immunodeficiency virus even though the virus was cultured from the peripheral blood lymphocytes. During follow-up for up to 60 weeks one cat died and 23 were destroyed on humane grounds.     

Analgesic, hemodynamic and respiratory effects of caudal epidurally administered ropivacaine hydrochloride in mares

Objective To determine the analgesic, hemodynamic and respiratory effects, sedation and ataxia in mares of caudal epidural administration of ropivacaine hydrochloride solution. Study design Prospective, single‐dose trial. Animals Ten healthy mares weighing from 475 to 565 kg. Methods Intravascular catheters and an epidural needle were placed after infiltration of the skin and subcutaneous tissues with 2% lidocaine. Ropivacaine (0.5%, 8 or 9 mL) was then injected epidurally at the fifth sacral or sacrococcygeal vertebrae, respectively. Analgesia was determined by lack of sensory perception to electrical stimulation (> 40 milliamps) and absence of response to needle pricks extending from coccyx to S2 dermatomes. Electrocardiogram, heart and respiratory rates, rectal temperature, arterial blood pressure, arterial acid‐base (pH, standard bicarbonate and base excess), gas tensions (PO₂, PCO₂), PCV, oxyhemoglobin and total solids concentrations, and numerical scores of perineal analgesia, sedation (head drop), and ataxia (position of pelvic limbs) were determined before and during a 5‐hour testing period. Analysis of variance (anova ) with repeated measures was used to detect significant (p < 0.05) differences of mean values from baseline. Results Epidurally administered ropivacaine induced variable analgesia extending bilaterally from coccyx to S2 (three mares), coccyx to S3 (four mares), and coccyx to S4 (three mares), with minimal sedation, ataxia, and cardiovascular and respiratory disturbances of mares. Perineal analgesia was attained at 10 ± 4 minutes and lasted for 196 ±42 minutes (mean ± SD). Five mares demonstrated inadequate perineal analgesia, probably attributable to deviation of the spinal needle from the midline. They were successfully blocked with ropivacaine on another occasion. Epidural ropivacaine significantly reduced repiratory rates of mares and did not change other variables from baseline. Conclusions and clinical relevance Ropivacaine (0.5%, 8 mL 500 kg^?1) can be administered caudal epidurally to produce prolonged (> 2.5 hours) bilateral perineal analgesia with minimal sedation, ataxia, and circulatory and respiratory disturbances in standing mares.     

Effect of sodium bicarbonate infusions on ionized calcium and total calcium concentrations in serum of clinically normal cats

The effects of sodium bicarbonate (0.5 mEq/kg of body weight, 1.0 mEq/kg, 2.0 mEq/kg, and 4.0 mEq/kg) on ionized and total calcium concentrations were determined in clinically normal cats. Also, serum pH, whole blood pH, and serum albumin, serum total protein, and serum phosphorus concentrations were measured. Intravenous administration of sodium bicarbonate to awake cats decreased serum ionized calcium and serum total calcium concentrations. All dosages of sodium bicarbonate were associated with significant decreases of serum ionized calcium concentration. This effect lasted for greater than 180 minutes when cats were given 2.0 mEq/kg or 4.0 mEq/kg. When cats were given 4 mEq of sodium bicarbonate/kg, serum ionized calcium concentration was significantly decreased, compared with that when cats were given lower doses, but only at 10 minutes after infusion. After sodium bicarbonate infusion, serum total calcium concentration, measured by ion-specific electrode and colorimetry, was lower than baseline values at most of the times evaluated. Decreases in serum ionized calcium and serum total calcium concentrations can be attributed only in part to an increase in serum or whole blood pH and to a decrease in serum protein concentration. Serum total calcium concentrations measured by ion-specific electrode and by colorimetry were positively correlated, but the variability was high. Only 44% of the variability in serum ionized calcium concentration could be predicted when serum total calcium, albumin, total protein, phosphorus, and bicarbonate concentrations and pH were considered.