The effects of different feeding strategies providing different levels of vitamin A on animal performance, carcass traits, and the conversion rate of subcutaneous fat color in cull-cows.

Cull cows represent a significant percentage of revenue received from the U.S. beef industry; however, cull cows are heavily price discounted at time of slaughter. This experiment's objective is to evaluate different feeding strategies and their effects on body condition score, subcutaneous fat color, and carcass yield and quality traits in cull cows. The central hypothesis is feeding a high-energy diet, with low levels of vitamin A, for 56 d will improve animal performance, carcass yield, and quality traits in addition to capturing the point (rate) of the conversion of yellow to white subcutaneous fat. In the present experiment 98 Angus crossbreed cows were utilized. Cows were fed either low vitamin A (LVA) diet consisting of whole shelled corn, soybean hulls, soybean meal, and a mineral-vitamin supplement or high vitamin A (HVA) diet, formulated using whole shelled corn, fescue hay, dry distiller grains with soluble, and a mineral-vitamin supplement for 56 d. During the 56 d feeding period, body weights and condition scores, and subcutaneous adipose samples were collected every 14 d. On day 56, cattle were slaughtered; 48 h postmortem carcass characteristics and objective color scores (subcutaneous adipose tissue) were recorded and a sample of the longissimus dorsi lumborum was collected. Subcutaneous adipose tissue samples were utilized to record subjective color scores and then ground to be analyzed for ß-carotene concentration. The longissimus dorsi lumborum samples (2.54 cm slices) were removed for Warner-Bratzler shear force (WBSF) and pH testing. Data were analyzed using the MIXED procedure of SAS. Feeding cull cows LVA resulted in differences in subcutaneous carcass fat color (P = 0.01) as well as b* values (P < 0.01) on day 56 compared with HVA. Subjective fat color scores were not different (P > 0.10) on day 0 or 14 but were different (P ≤ 0.05) on days 28, 42, and 56. Additionally, 9-cis-ß-carotene concentration on day 56 were different (P = 0.05) between treatments. A trend was noticed for all-trans-ß-carotene concentration (P = 0.10) on day 56 as well. Cull cow body weights were greater (P ≤ 0.04) when fed the LVA diet starting on days 14, 28, and 42; and a trend was noticed on day 56 (P = 0.09). Overall, cows fed the LVA treatment for 56 d exhibited decreased adipose yellowness and ß-carotene concentrations as well as increased live weights.

Reduced embryo yield obtained from superstimulated ewes with low circulating AMH concentration is improved by lengthening the FSH treatment.

The aim of the present study was to evaluate: 1) the association between AMH, AFC, superovulatory response and embryo yield in sheep; and 2) the effect of FSH treatment length during superstimulation of the first follicular wave on ovarian response and embryo yield, particularly in ewes with low and high AMH. The experiment was performed on 63 Polled Dorset ewes that received an ovarian superstimulatory treatment during the first follicular wave (Day 0 protocol). Ewes were administered a total dose of 240 mg of FSH distributed in six (6-dose regimen, n = 30) or eight (8-dose regimen, n = 33) decreasing doses administered 12 h apart. On Day -9 (random stage of the estrous cycle) and Day 0 (day of the first FSH dose) ovarian ultrasonography was performed and blood samples were collected for AFC and AMH determinations, respectively. A weak positive correlation between AMH and small AFC (follicles <4 mm) was observed (r = 0.23; P = 0.07), and AMH concentration was positively correlated (r = 0.29; P < 0.05) with the number of corpora lutea (CL) determined at embryo collection (i.e., 6 d after insemination). The length of FSH treatment tended (P = 0.06) to affect the ovarian response, such that the number of CL was greater in 8-dose than 6-dose treated ewes, while no differences (P > 0.10) in embryo yield outcomes were observed. For further analysis, ewes were classified into low (<7 ng/mL) and high (>10 ng/mL) serum AMH. In high AMH ewes, there were no differences (P > 0.05) in the number of CL nor embryo yield between the 6-dose and 8-dose treatment (e.g., 7.8 ± 2.4 and 8.3 ± 2.5 transferable embryos, respectively; P = 0.92). Conversely, for low AMH ewes, fertilized ova and embryo yield were greater (P ≤ 0.05) for ewes receiving the 8-dose than the 6-dose superstimulatory treatment (e.g., 8.4 ± 2.8 vs. 2.7 ± 0.9 transferable embryos, respectively, P ≤ 0.05). In conclusion, embryo production in poor responding ewes with low low circulating AMH is improved by extending the superstimulatory treatment length from 6 to 8 FSH doses.

GnRH dose at initiation of a 5-day CO-Synch + P4 for fixed time artificial insemination in suckled beef cows.

The objective of the present study was to evaluate the effect of GnRH dose administered at initiation (GnRH-1) of a 5-day CO-Synch + P4 protocol on ovulatory response, expression of estrus, and fertility in suckled beef cows. Suckled beef cows (n = 1101) at four locations were randomized to receive either 100 or 200 µg of gonadorelin acetate at initiation (D-8) of a 5-day CO-Synch + P4 protocol concurrently with insertion of an intravaginal progesterone (P4) device. On D-3 the P4 device was removed, two doses of prostaglandin F2α were administered concurrently and a patch was applied to evaluate expression of estrus. Artificial insemination was performed 72 h after P4 device removal (D0) simultaneously with the administration of 100 µg of gonadorelin acetate (GnRH-2). Increasing GnRH dose at initiation of a 5-day CO-Synch + P4 did not enhance ovulatory response (P = 0.57) to GnRH-1, expression of estrus (P = 0.79), nor pregnancies per AI (P/AI; P = 0.91). Both follicle size (quadratic) and circulating P4 (linear) affected (P < 0.01) ovulatory response to GnRH-1 independent of dose. Cows that had ovulation to GnRH-1 had smaller (P < 0.001) follicle size on D-3 and reduced (P = 0.05) expression of estrus compared to cows that did not have ovulation to GnRH-1, however, P/AI did not differ (P = 0.75). In conclusion, increasing the dose of GnRH-1 in the 5-day CO-Synch + P4 protocol did not enhance ovulatory response, expression of estrus, or P/AI in suckled beef cows.

Atlantic sturgeon and shortnose sturgeon exhibit highly divergent transcriptomic responses to acute heat stress.

In comparison to most modern teleost fishes, sturgeons generally display muted stress responses. While a muted stress response appears to be ubiquitous across sturgeon species, the mechanisms unpinning this muted response have not been fully described. The objective of this study was to determine the patterns of hematological and transcriptomic change in muscle tissue following an acute high temperature stress (critical thermal maxima; CTmax) in two locally co-occurring but evolutionarily distant sturgeon species (Atlantic and shortnose sturgeon). The most striking pattern found was that Atlantic sturgeon launched a vigorous transcriptomic response at CTmax, whereas shortnose sturgeon did not. In contrast, shortnose sturgeon have significantly higher cortisol than Atlantics at CTmax, reconfirming that shortnose have a less muted cortisol stress response. Atlantic sturgeon downregulated a number of processes, included RNA creation/processing, methylation and immune processes. Furthermore, a number of genes related to heat shock proteins were differentially expressed at CTmax in Atlantic sturgeon but none of these genes were significantly changed in shortnose sturgeon. We also note that the majority of differentially expressed genes of both species are undescribed and have no known orthologues. These results suggest that, while sturgeons as a whole may show muted stress responses, individual sturgeon species likely use different inducible strategies to cope with acute high temperature stress.

Short communication: pharmacokinetics of oxytocin administered intranasally to beef cattle.

Providing the neuropeptides oxytocin and vasopressin intranasally increased concentrations in plasma and cerebral spinal fluid in humans and primates, respectively. This is of interest because of the documented anxiolytic effects of oxytocin observed in humans and rodents. To date, a transnasal approach of hormone administration has not been investigated in beef cattle. Defining the pharmacokinetics of intranasal oxytocin in cattle is necessary for determining optimum sampling and dosing timelines for future investigations. Five, weaned Bos taurus steers were used in a 3 × 3 Latin square design. Treatments included 1) 0.33 IU oxytocin/kg BW (A, n = 5), 2) 0.66 IU oxytocin/kg BW (B, n = 5), and 3) 1.32 IU oxytocin/kg BW (C, n = 5). Steers were acclimated to handling and restraint procedures for 4 wk leading up to the start of the experiment. Frequent blood collection occurred every 2 min for the first 30 min and every 5 min for the second 30 min, relative to administration of intranasal treatment. No treatment by time interaction was detected; however, there was an effect of time (P < 0.001) and treatment (P = 0.002) on oxytocin concentrations over time. Pharmacokinetic parameters, determined by PKSolver excel add-in, demonstrated an average maximum concentration (CMAX) of 63.3 pg/mL at 3.5 min after intranasal dose administration. An average half-life (T1/2) of 12.1 min after intranasal administration was determined. Pharmacokinetic parameters to a single bolus were not dose-dependent.

Intranasal oxytocin treatment does not attenuate the hypothalamo-pituitary-adrenal axis in beef heifers subjected to isolation stress or restraint and isolation stress.

Changes in the physiological, psychological, and behavioral manifestations of stress have been observed in association with increases in circulating oxytocin (OXT). Providing OXT intranasally has been shown to attenuate stressor-induced hypothalamo-pituitary-adrenal (HPA) axis activation in humans and rodents; however, anxiolytic effects may be context and species specific. The present study aimed to investigate the effect of intranasal OXT supplementation on stressor-induced activation of the HPA axis in beef cattle. We hypothesized that OXT would attenuate activation of the HPA axis, ultimately decreasing plasma cortisol and adrenocorticotropic hormone (ACTH). Twenty-eight Bos taurus heifers were blocked by bodyweight and randomly allocated to one of four treatment groups, in a 2 × 2 factorial arrangement: (1) saline, isolated, standing, and unrestrained (S-isolation stress [IS], 0.015 mL/kg BW 0.9% isotonic saline, n = 7); (2) saline, isolated, and restrained (S-restraint and isolation stress [RIS]; 0.015 mL/kg BW 0.9% isotonic saline; n = 7); (3) OXT, IS (OXT-IS, 0.3 IU/kg BW oxytocin; n = 7); and (4) OXT and RIS (OXT-RIS, 0.3 IU/kg BW oxytocin; n = 7). Oxytocin and saline were administered intranasally. Intranasal treatments were given followed by a waiting time of 30 min when each of the stress treatments was applied for 2 h. Blood samples were collected via jugular catheters directly after stressor application and every 10 min thereafter, for 2 h. Cortisol concentrations increased over time in animals exposed to RIS (P < 0.01) and decreased over time in animals exposed to IS (P < 0.01). Concentrations of ACTH decreased over time for the IS-treated heifers but remained elevated for the RIS-treated heifers (P < 0.01). Under the conditions of the present study, OXT treatment did not affect measured indicators of HPA axis activation. A treatment × time interaction (P < 0.01) was detected for OXT, such that OXT heifers exhibited greater initial OXT concentrations followed by a decline; saline-treated heifers had consistently stable oxytocin concentrations. The RIS-treated heifers increased their glucose (P < 0.01) and lactate (P < 0.01) concentrations throughout the application of the stressors compared with the IS-treated heifers. Overall, restraint stress increased cortisol and oxytocin in B taurus heifers compared with heifers subjected only to isolation. Finding a more intermediate stress model may better allow for detection of the effects of oxytocin on the stress response.

Lack of change in swimming capacity (Ucrit) following acute salinity exposure in juvenile shortnose sturgeon (Acipenser brevirostrum).

The Saint John River (SJR) is home to the only Canadian population of shortnose sturgeon, Acipenser brevirostrum. Adult shortnose sturgeon routinely enter saltwater to forage, yet less is known about how juveniles cope with the associated osmoregulatory pressures. Recently, it has been shown that short-term (24 h) exposure to saltwater causes significant changes to ion and water levels in juvenile shortnose. In some species of fish, notably salmonids, it has been shown that shifts in fluid and ion levels following saltwater challenges reduce the swimming capacity. The relationship between ion concentration and swimming capacity is not well understood for sturgeon species. Our research aimed to determine whether short-term salt exposure affects swimming ability in juvenile shortnose sturgeon. Juvenile, SJR, hatchery-raised shortnose sturgeon (< 1 year old) were exposed to salinities of 0 (control), 16, or 24‰ for 24 h and then subjected to a critical swimming speed test (Ucrit) to quantify swimming ability. Following the test, the fish were weighed and blood samples were drawn to be analyzed for plasma ion and cortisol levels. While ion levels and weight loss were significantly higher in salt exposed fish, there were no significant differences in critical swimming speed or cortisol concentrations. This is in contrast to what has been observed in salmonids and Adriatic sturgeon. This suggests the hydromineral imbalance caused by moderate salt exposure is not sufficient to affect the swimming performance of shortnose sturgeon. Shortnose sturgeon are not thought to enter the saline stretches of the SJR until roughly 8 years of age, yet this research shows that much younger juveniles withstand moderate salinity for short periods, with little whole-animal ramifications.

The effect of substratum type on aspects of swimming performance and behaviour in shortnose sturgeon Acipenser brevirostrum.

The swimming performance and associated swimming behaviour (i.e. substratum-skimming, station-holding and free swimming) were assessed in shortnose sturgeon Acipenser brevirostrum during critical swimming and endurance swimming tests over a rough and a smooth substratum. It was hypothesized that the addition of a rough substratum in the swimming flume may provide a surface for the A. brevirostrum to grip and offer an energetic advantage. Substratum type did not affect the critical swimming performance, but A. brevirostrum consistently performed more bottom behaviours (i.e. substratum-skimming and station-holding) while on a smooth substratum. Acipenser brevirostrum had little contact with the rough substratum until the velocity was >1 body length s-1 . Endurance swimming time was significantly lower for A. brevirostrum over the rough bottom at the highest velocity (30 cm s-1 ) which may be attributed to the observed increase in free swimming and decrease in bottom behaviours. During endurance swimming, the rough substratum was mainly used at intermediate velocities, suggesting that there may be a stability cost associated with being in contact with the rough substratum at certain velocities.

Oxygen consumption and haematology of juvenile shortnose sturgeon Acipenser brevirostrum during an acute 24 h saltwater challenge.

This study focused on the acute physiological responses to saltwater exposure in juvenile shortnose sturgeon Acipenser brevirostrum. In two separate laboratory experiments, 2 year-old A. brevirostrum were exposed to either full (32) or half-strength (16) seawater for up to 24 h. First, oxygen consumption rates were used to estimate the metabolic costs over 24 h. Secondly, blood and muscle samples were analysed at 6, 12 and 24 h for water loss, various measures of osmoregulatory status (plasma osmolality and ions) and other standard haematological variables. Juveniles exposed to full-strength seawater showed significant decreases in oxygen consumption rates during the 24 h exposure. Furthermore, seawater-exposed fish had significantly increased plasma osmolality, ions (Na(+) and Cl(-)) and a 17% decrease in total wet mass over the 24 h exposure period. To a lesser extent, increases in osmolality, ions and mass loss were observed in fish exposed to half-strength seawater but no changes to oxygen consumption. Cortisol was also significantly increased in fish exposed to full-strength seawater. While plasma protein was elevated following 24 h in full-strength seawater, haemoglobin, haematocrit and plasma glucose levels did not change with increased salinity. These results imply an inability of juvenile A. brevirostrum to regulate water and ions in full-strength seawater within 24 h. Nonetheless, no mortality occurred in any exposure, suggesting that juvenile A. brevirostrum can tolerate short periods in saline environments.

The influence of flume length and group size on swimming performance in shortnose sturgeon Acipenser brevirostrum.

The main objectives of this study were to determine optimal methodologies to assess the general swimming performance of juvenile shortnose sturgeon Acipenser brevirostrum. Swimming densities (group v. individual swimming) and flume length (2 v. 1 m) were altered to verify if any of those variables affected performance (i.e. time to fatigue) during critical swimming (U(crit)) and endurance tests. Results for both U(crit) and endurance swimming were not significantly different between fish swum in groups of five or fish swum individually. The U(crit) values, however, were c. 22% higher for fish swum in a longer flume. Although swimming fish in groups did not improve swimming performance, group swimming lowered the variance of the data. Results also reveal that juvenile A. brevirostrum may not possess an ability to swim at high speeds (i.e. burst phase) for long periods.

Oxygen consumption, ammonia excretion and protein use in response to thermal changes in juvenile Atlantic salmon Salmo salar.

Experiments were designed to examine the effects of various temperature challenges on oxygen consumption and ammonia excretion rates and protein utilization in juvenile Atlantic salmon Salmo salar. Fish acclimated to 15 degrees C were acutely and abruptly exposed to either 20 or 25 degrees C for a period of 3 h. To simulate a more environmentally relevant temperature challenge, a third group of fish was exposed to a gradual increase in temperature from 15 to 20 degrees C over a period of 3 h (c. 1.7 degrees C h(-1)). Oxygen consumption and ammonia excretion rates were monitored before, during and after the temperature shift. From the ammonia excretion and oxygen consumption rates, protein utilization rates were calculated. Acute temperature changes (15-20 degrees C or 15-25 degrees C) caused large and immediate increases in the oxygen consumption rates. When the temperature was gradually changed (i.e. 1.7 degrees C h(-1)), however, the rates of oxygen consumption and ammonia excretion were only marginally altered. When fish were exposed to warmer temperatures (i.e. 15-20 degrees C or 15-25 degrees C) protein use generally remained at pre-exposure (15 degrees C) levels. A rapid transfer back to 15 degrees C (20-15 degrees C or 25-15 degrees C) generally increased protein use in S. salar. These results indicate that both the magnitude and the rate of temperature change are important in describing the physiological response in juvenile salmonids.

Behaviour and performance of juvenile shortnose sturgeon Acipenser brevirostrum at different water velocities.

Critical swimming speeds (mean +/-s.e.) for juvenile shortnose sturgeon Acipenser brevirostrum were 34.4 cm s(-1)+/- 1.7 (2.18 +/- 0.09 body lengths, BL s(-1)). Swimming challenges at 10, 20 and 30 cm s(-1) revealed that juvenile A. brevirostrum are relatively poor swimmers, and that the fish did not significantly modify their swimming behaviour, although they spent more time substratum skimming (i.e. contact with flume floor) at 30 cm s(-1) relative to 10 cm s(-1). When present, these behavioural responses are probably related to morphological features, such as flattened rostrum, large pectoral fins, flattened body shape and heterocercal tail, and may be important to reduce the costs of swimming.

Sub-lethal ammonia toxicity in largemouth bass.

Guidelines for ammonia toxicity in fish are often determined using static exposure tests with immature fish over a 96-h period. These results may not be relevant to aquaculture, hauling or angling tournament scenarios where mature fish can be exposed to ammonia for shorter durations, often following additional stressors such as handling. The current study sought to quantify (1) the impact of ambient ammonia on the ability of largemouth bass to recover from exercise, (2) the behavioural response of largemouth bass to elevated ambient ammonia and (3) the concentration of ammonia that can accumulate in a live-release vessel at an angling tournament. After approximately 3 h, total ammonia (T(amm)) concentrations in a live-release vessel at an angling tournament were almost 200 muM. Exposure of fish to 1000 microM T(amm) (a value approximately 80% below the criteria maximum concentration for largemouth bass) caused significant reductions in ventilation rates, and increases in erratic swimming and irregular ventilation. Exposure to 100 microM T(amm) impaired the ability of largemouth bass to recover from exercise relative to fish recovering in fresh water. Therefore, sub-lethal ambient ammonia concentrations cause physiological disturbances that can impair the recovery of largemouth bass from exercise.

Juvenile atlantic and shortnose sturgeons (family: Acipenseridae) have different hematological responses to acute environmental hypoxia.

Experiments were conducted to determine the behavioral and physiological responses to acute hypoxic challenges in Atlantic (Acipenser oxyrinchus) and shortnose (Acipenser brevirostrum) sturgeons. We measured the ventilatory rate following a 45-mmHg hypoxic challenge, as well as a variety of hematological parameters, including O2 transport and hormonal, ionic, and metabolic variables, following a 1-h exposure to either 75- or 30-mmHg hypoxic challenges. Compared to fish in normoxic conditions (Pwo2 150 mmHg), juveniles of both species increased their ventilatory rate by approximately 40% when exposed to a 1-h challenge at 45 mmHg Pwo2. Hematological variables (e.g., hematocrit, hemoglobin, and Na+ and Cl- levels) did not change substantially following a 1-h challenge at 75 mmHg Pwo2. Conversely, a severe hypoxic challenge of 30 mmHg caused changes in several hematological variables (e.g., whole blood glucose and plasma cortisol and lactate levels). Most of these hematological parameters returned to prehypoxic levels within 2 h. Severe environmental hypoxia elicited the same basic pattern of response in both species; however, maximal plasma lactate levels were higher in Atlantic sturgeons, and maximal cortisol levels were higher in shortnose sturgeons. Whether these species differences are related to dissimilar hypoxia-tolerance, ecological, and/or endocrinological characteristics between these two species is not entirely clear.

Effects of metal mining effluent on Atlantic salmon (Salmo salar) and slimy sculpin (Cottus cognatus): using artificial streams to assess existing effects and predict future consequences.

In the summer of 2000, the effects of metal mine discharge on fish growth and exercise performance were assessed at a Zn-Pb-Cu mine in New Brunswick, Canada. Juvenile Atlantic salmon (Salmo salar) were exposed to 0%, 20%, and 80% treated metal mine effluent in a mobile, fish-only artificial stream system. Fish were fed commercial salmon pellets throughout the study. Young-of-the-year slimy sculpins (Cottus cognatus) were exposed to the same treatments in a multitrophic level, modular artificial stream system or mesocosm, in which the fish were dependent on seeded algae and invertebrates for nutrition. Treatment concentrations were chosen to represent existing discharge dilutions (80%) and a scenario of reduced effluent discharge (20%) as predicted upon mine closure (scheduled for 2008). Al, Ba, B, Fe, Mn, Sr, Tl, Ti, and Zn increased in a concentration-dependent fashion across the three treatments. Salmon body burdens of Ba, Cd, Li, Cu, Mn, Se, Sr, and Zn were increased in the 80% treatment, while Tl increased across all treatment levels. Mortalities and depressions in growth in both fish species paralleled treatment concentrations (80%>20%>0%). Salmon liver weight was significantly greater in fish exposed to 20% and 80% effluent in a concentration-dependent fashion. Exercise performance in fish, as assessed by the ability to recover from forced exercise, showed little effect of treatment. The contamination of the receiving environment by mine discharges has led to loss of fish, making it impossible to study the system in situ. However, the use of the artificial stream systems enabled us to assess effects of present conditions on fish, as well as the potential impacts of mine reclamation. The 20% discharge predicted following mine reclamation is potentially favourable for the reinstitution of native fishes into the system.

The physiological response of diploid and triploid brook trout to exhaustive exercise.

Using triploidy as an experimental model, we examined whether cell size limits the post-exercise recovery process in fish. Because triploids generally possess larger cells, which could affect many physiological and biochemical processes, we hypothesized that triploids would take longer to recover from exhaustive exercise compared to diploids. To test this, we measured plasma lactate, glucose and osmolality, and white muscle energy stores (glycogen, phosphocreatine and ATP) and lactate before and immediately following exhaustive exercise and during recovery at 2 and 4 h post-exercise. In addition, oxygen consumption and ammonia excretion rates were determined before and after exhaustive exercise. Overall, diploid and triploid brook trout showed similar metabolic responses exercise, but plasma osmolality, white muscle lactate, white muscle ATP and post-exercise oxygen consumption rates recovered earlier in triploids compared to diploids. The results of this study suggest that the characteristic larger cell size of triploidy does not limit the physiological response to, or recovery from, exhaustive exercise.

Tonsillar B cells do not express PSGL-1, but a significant fraction displays the cutaneous lymphocyte antigen and exhibits effective E- and P-selectin ligand activity.

Skin-homing T cells are defined by the expression of the cutaneous lymphocyte-associated antigen (CLA) which enables the cells to selectively bind to vascular endothelial E-selectin close to sites of cutaneous inflammation, an initial step in the effective extravasation from blood into the inflamed tissue. Essentially all CLA on T cells decorates the backbone of the P-selectin glycoprotein ligand-1 (PSGL-1). In this study we show that human peripheral blood B cells (PBBC) and tonsillar B cells (TBC) do not display PSGL-1 in fluorescence-activated cell sorter analysis using different murine monoclonal antibodies and polyclonal rabbit anti-PSGL-1 antiserum. A significant population of TBC, however, expresses a HECA-452-reactive epitope. These cells represent nonactivated IgM(+)/IgG(-) mature B lymphocytes. Up to 50% of the TBC in a given preparation strongly bind to E- and up to 79% to P-selectin. The shear stress resistance in a parallel-plate flow chamber system was high. Neuraminidase treatment of TBC totally and O-sialoglycoprotein endopeptidase partially diminished HECA-452 reactivity and reduced E- but not P-selectin ligand activities. Mocarhagin had no effect in the assays. The data suggest a different ligand for P-selectin and a distinct glycoprotein carrier for the E-selectin ligand as compared to T cells or other leukocytes. Adhesion to P-selectin, however, still required sulfation of the ligand for function. Western blots of TBC cell lysates detected a >240-kD HECA-452-reactive material that was resistant to reducing conditions. Anti-PSGL-1 did not reveal immunoreactive material in these cell lysates. B cell activation did neither significantly change HECA positivity nor induce PSGL-1 expression. Cultured, activated TBC, however, maintained expression of the integrin alpha4beta7. Human peripheral blood B cells had similar cell surface characteristics to TBC. Our observations suggest that several adhesion molecules may be involved in B cell homing which include CLA, the P-selectin ligand, and structures such as alpha4beta7.

Inflammatory skin disease in K14/p40 transgenic mice: evidence for interleukin-12-like activities of p40.

The proinflammatory cytokine interleukin-12, a p35/p40 heterodimer, is produced by resident cells in skin and has been implicated as a pathogenetic factor in T-cell-mediated skin diseases. Secretion of heterodimeric interleukin-12 is always accompanied by production of p40 monomer and p40/p40 homodimer. To investigate the possible in vivo role of p40 per se, we generated mice that constitutively express monomeric and homodimeric p40 in basal keratinocytes. These mice spontaneously developed an eczematous skin disease that was characterized by hyperkeratosis, focal epidermal spongiosis, and a mixed inflammatory infiltrate composed of T cells (CD4+), macrophages, eosinophils, mast cells, and few neutrophils. Fluorescence-activated cell sorter analysis of transgenic epidermal cell suspensions revealed induction of major histocompatibility complex class II molecules on keratinocytes and a 2-3-fold increase in the content of Langerhans cells. Cytokines produced by these activated epidermal cells include interleukin-1alpha and tumor necrosis factor alpha. The skin disease in K14/p40 mice was similar to that of littermate mice that received injections of interleukin-12, suggesting overlapping in vivo functional properties. As induction of interferon-gamma is a major function of interleukin-12, we tested the in vitro ability of transgenic p40 to induce interferon-gamma. In contrast to interleukin-12, transgenic p40 did not stimulate interferon-gamma secretion by cultured splenocytes. We conclude that transgenic p40 and interleukin-12 are equally capable of initiating cutaneous inflammation. Despite these in vivo similarities, there is a clear functional difference between interleukin-12 and transgenic p40 in vitro, suggesting that interferon-gamma is not a major factor contributing to interleukin-12-like activities of transgenic p40.

Neutrophils, monocytes, and dendritic cells express the same specialized form of PSGL-1 as do skin-homing memory T cells: cutaneous lymphocyte antigen.

Memory T cells in inflamed skin express the cutaneous lymphocyte-associated antigen (CLA), a glycosylated epitope defined by the mAb HECA-452. We previously reported that on T cells, CLA occurs almost exclusively on the protein backbone of P-selectin glycoprotein ligand-1 (PSGL-1). T cells exhibiting the CLA isoform of PSGL-1 can tether and roll on both E- and P-selectin, while T cells expressing PSGL-1 without the CLA epitope do not bind E-selectin, though they may bind P-selectin. We show here that circulating neutrophils and monocytes, and cultured blood dendritic cells, also express CLA almost entirely as an isoform of PSGL-1. These cells all tether and roll on both E- and P-selectin. A chimeric fusion protein incorporating the 19 N-terminal amino acids of mature PSGL-1 exhibited HECA-452 immunoreactivity and supported rolling of CHO cells expressing either E- or P-selectin. These findings indicate a site for the CLA modification within the distal tip of PSGL-1, previously shown to be critical for P-selectin binding and to mediate some, but not all, of the E-selectin binding of PSGL-1. We hypothesize that the types of circulating leukocytes discussed above all use CLA/PSGL-1 to tether and roll on E- and P-selectin along the vascular endothelium.

Juvenile sturgeon exhibit reduced physiological responses to exercise.

Experiments were conducted to determine the physiological responses to exercise of Atlantic sturgeon (Acipenser oxyrhynchus) and shortnose sturgeon (A. brevirostrum). We measured the rates of oxygen consumption and ammonia excretion in both species and a variety of physiological parameters in both muscle (e.g. lactate, glycogen, pyruvate, glucose and phosphocreatine concentrations) and blood (e.g. osmolality and lactate concentration) in juvenile shortnose sturgeon following 5 min of exhaustive exercise. In both species, oxygen consumption and ammonia excretion rates increased approximately twofold following exhaustive exercise. Post-exercise oxygen consumption rates decreased to control levels within 30 min in both sturgeon species, but post-exercise ammonia excretion rates remained high in Atlantic sturgeon throughout the 4 h experiment. Resting muscle energy metabolite levels in shortnose sturgeon were similar to those of other fish species, but the levels decreased only slightly following the exercise period and recovery occurred within an hour. Under resting conditions, muscle lactate levels were low (<1 micromol g(-1)) but they increased to approximately 6 micromol g(-1) after exercise, returning to control levels within 6 h. Unlike similarly stressed teleost fish, such as the rainbow trout, plasma lactate levels did not increase substantially and returned to resting levels within 2 h. Plasma osmolality was not significantly affected by exercise in shortnose sturgeon. Taken together, these results suggest that shortnose and Atlantic sturgeon do not exhibit the physiological responses to exhaustive exercise typical of other fish species. They may possess behavioural or endocrinological mechanisms that differ from those of other fishes and that lead to a reduced ability to respond physiologically to exhaustive exercise.