Use of monoclonal antibodies against human HLA II antigens for the detection of bovine B lymphocytes and macrophages]

The crossreactivity of mouse monoclonal antibodies (MoAbs) (Tab. I) prepared against human HLA-DR and HLA-DP antigens was studied in various bovine cells: lymphocytes from lymph nodes and peripheral blood, adherent (B) and nonadherent (T) lymphocytes, monocytes, granulocytes and platelets. In the immunofluorescence test, MoAbs Bra13, Bra14, Bra20, Bra22, Bra30, Bra70, HL-38 reacted with bovine B lymphocytes and monocytes, but not with other tested cells (Tab. III, IV). These antibodies, except Bra22, were positive with B lymphocytes in the complement dependent cytotoxic test (Tab. II). The similarity of the bovine antigens and HLA-DR antigens determined by used MoAbs was also proved by immunoblotting. Monoclonal antibodies Bra38 and BraFB6 did not react with the bovine cells and separated antigens. The epitope (HLA-DR) recognized by the antibody Bra38 is probably absent in cattle. The presence of HLA-DP analogue determined by the antibody BraFB6 has not been confirmed. The crossreactive MoAbs could be used for the detection of B lymphocytes and macrophages in veterinary immunology.     

Stomatal response of an anisohydric grapevine cultivar to evaporative demand, available soil moisture and abscisic acid

Stomatal responsiveness to evaporative demand (air vapour pressure deficit (VPD)) ranges widely between species and cultivars, and mechanisms for stomatal control in response to VPD remain obscure. The interaction of irrigation and soil moisture with VPD on stomatal conductance is particularly difficult to predict, but nevertheless is critical to instantaneous transpiration and vulnerability to desiccation. Stomatal sensitivity to VPD and soil moisture was investigated in Semillon, an anisohydric Vitis vinifera L. variety whose leaf water potential (Ψ(l)) is frequently lower than that of other grapevine varieties grown under similar conditions in the warm grape-growing regions of Australia. A survey of Semillon vines across seven vineyards revealed that, regardless of irrigation treatment, midday Ψ(l) was dependent on not only soil moisture but VPD at the time of measurement. Predawn Ψ(l) was more closely correlated to not only soil moisture in dry vineyards but to night-time VPD in drip-irrigated vineyards, with incomplete rehydration during high night-time VPD. Daytime stomatal conductance was low only under severe plant water deficits, induced by extremes in dry soil. Stomatal response to VPD was inconsistent across irrigation regime; however, in an unirrigated vineyard, stomatal sensitivity to VPD-the magnitude of stomatal response to VPD-was heightened under dry soils. It was also found that stomatal sensitivity was proportional to the magnitude of stomatal conductance at a reference VPD of 1kPa. Exogenous abscisic acid (ABA) applied to roots of Semillon vines growing in a hydroponic system induced stomatal closure and, in field vines, petiole xylem sap ABA concentrations rose throughout the morning and were higher in vines with low Ψ(l). These data indicate that despite high stomatal conductance of this anisohydric variety when grown in medium to high soil moisture, increased concentrations of ABA as a result of very limited soil moisture may augment stomatal responsiveness to low VPD.     

Hematopoietic cell regulation by Rac1 and Rac2 guanosine triphosphatases

The Rho guanosine triphosphatases (GTPases) Rac1 and Rac2 are critical signaling regulators in mammalian cells. The deletion of both Rac1 and Rac2 murine alleles leads to a massive egress of hematopoietic stem/progenitor cells (HSC/Ps) into the blood from the marrow, whereas Rac1-/- but not Rac2-/- HSC/Ps fail to engraft in the bone marrow of irradiated recipient mice. In contrast, Rac2, but not Rac1, regulates superoxide production and directed migration in neutrophils, and in each cell type, the two GTPases play distinct roles in actin organization, cell survival, and proliferation. Thus, Rac1 and Rac2 regulate unique aspects of hematopoietic development and function.     

Littoral spawning habitats of three southern Arctic charr (Salvelinus alpinus L.) populations

Abstract Arctic charr populations in southern latitudes are nonmigratory, with all life‐stages limited to freshwater lakes and in‐ or out‐flowing tributaries. Although many of these populations are reported to also spawn in lake littorals, little is known about the physical characteristics of putative spawning grounds. A total of 23 discrete spawning sites within three Irish lakes were located by fyke netting of spawning adults and snorkelling in littoral habitats. Spawning sites were found to be long, narrow strips running parallel to the shore at a maximum depth of 124 cm. Spawning sites were limited to areas of coarse mineral substrate with an adequate (c. 8 cm) depth of clean interstitial spaces. In individual lakes, combined areas of spawning sites made up 0.4–0.7% of available littoral. Egg densities varied considerably between sites (33–900·eggs m^?2) and were significantly correlated with gradient and width of spawning sites. No evidence of redd digging was found. The shallow, localised and restricted nature of spawning grounds makes such populations vulnerable to anthropogenically induced postoviposition changes in surface water level, eutrophication processes such as increased lake sedimentation and elevated nutrient status.     

Cryopreservation of trochophore larvae from the hard clam Mercenaria mercenaria: Evaluation of the cryoprotectant toxicity,cooling rate and thawing temperature

Aquaculture of hard clams Mercenaria mercenaria is a $65 million industry along the east coast and Gulf of Mexico coast in the United States. The goal of this study was to develop a preliminary protocol to cryopreserve trochophore larvae of hard clams. The objectives were to evaluate the: 1) toxicity of cryoprotectants, including dimethyl sulfoxide (DMSO), propylene glycol, ethylene glycol and glycerol, at 5, 10, 15 and 20% for exposure time of 1, 15, 30, 45, 60 and 75 min; 2) effects of cooling rates (5, 10, 20 and 30°C/min for the first trial; and 1, 3 and 5°C/min for second trial from 4 to ?80°C), thawing temperature (30, 40 and 50°C) and their interactions on post‐thaw viability. A basic protocol was concluded as: 15‐hr trochophore larvae mixed with DMSO or propylene glycol (5, 10%), equilibrated for 15 min, cooled in a programmable freezer from 4 to ?80°C at a cooling rate of 5°C/min and thawed at 50°C for 6 s. With this protocol, the immediate post‐thaw trochophore survival was 23 ± 14%, and survival to D‐stage was 27 ± 14%. This is the first report on larval cryopreservation in the hard clam and would have application for genetic breeding and seed production.     

Effects of atmospheric ammonia on pulmonary bacterial clearance in the young pig.

Young pigs were exposed to an aerosol of a nonpathogenic strain of Escherichia coli and then were retained in air-pollutant exposure chambers for a 2-hour clearance period. In series 1 (n = 80 pigs), 40 exposed young pigs (principals; 15.5 days of age) were placed in an atmosphere of filtered room air + 50 ppm of atmospheric NH3 during the clearance period; control pigs were exposed to filtered room air without added NH3. In series 2 (n = 24 pigs), 12 exposed young pigs (principals; 6.2 days of age) were similarly maintained, but at a lower concentration of atmospheric NH3 (75 ppm). At the end of the clearance period pigs were killed and pulmonary bacterial clearance was determined. Pigs kept in the NH3-contaminated atmospheres (either concentration) harbored more bacteria, on the average, in their lungs than did the controls. If series 1 and 2 data were combined, pigs kept in the NH3-contaminated atmospheres had 51% more bacteria in their lungs than did the controls. Pulmonic weight and ratio of pulmonic weight to body weight of pigs kept in the NH3-contaminated atmosphere were greater than those of the controls in series 1, but not in series 2. Gross and histopathologic examinations of lung tissue generally revealed no differences between controls and principals in either series 1 or 2.     

The analgesic and sedative effects of GV20 pharmacopuncture with low-dose hydromorphone in healthy dogs undergoing ovariohysterectomy

This study evaluates the analgesic efficacy of low-dose hydromorphone administered via pharmacopuncture at Governing Vessel 20 (GV20) for postoperative pain management following canine ovariohysterectomy. Fifty clinically healthy female dogs undergoing ovariohysterectomy were allocated to receive hydromorphone [0.1 mg/kg body weight (BW)] intramuscularly (IM, n = 25) or hydromorphone (0.01 mg/kg BW) pharmacopuncture at GV20 (GV, n = 25) following extubation. This was a prospective, blinded, randomized clinical trial. Pain and sedation scores were evaluated using the Glasgow Composite Measure Pain Scale Short Form (CMPS-SF) at 1, 2, 3, 4, and 12 hours following study treatment. Time of treatment failure (CMPS-SF ≥ 6/24) was recorded and analyzed using Kaplan-Meier survival analysis. Patient demographics and duration of surgery and anesthesia were analyzed using the appropriate unpaired Student’s t-test. The Glasgow CMPS-SF and sedation score were analyzed using a repeated measures 2-way analysis of variance (ANOVA) followed by Bonferroni post-test where appropriate. Significance was set a P < 0.05. There were no significant differences in patient demographics, anesthesia and surgery duration, and study treatment failure. The Glasgow CMPS-SF scores were significantly higher for IM compared with GV [2 (0 to 8) versus 1 (0 to 6), respectively; P = 0.044] at 4 hours. Sedation scores were significantly higher for IM compared with GV at 2 [2 (1 to 3) and 1 (1 to 3), respectively; P = 0.0004] and 4 [1 (1 to 3) and 1 (1 to 2), respectively; P = 0.03] hours. Pharmacopuncture with low-dose hydromorphone provided adequate postoperative analgesia in dogs undergoing ovariohysterectomy with reduced sedative effects. Pharmacopuncture is a good alternative in dogs when reduced dosing of opioids is recommended.     

Low levels of haptoglobin and putative amino acid sequence in Taiwanese Lanyu miniature pigs

Porcine haptoglobin (Hp) is an acute phase protein. Its plasma level increases significantly during inflammation and infection. One of the main functions of Hp is to bind free hemoglobin (Hb) and inhibit its oxidative activity. In the present report, we studied the Hp phenotype of Taiwanese Lanyu miniature pigs (TLY minipigs; n=43) and found their Hp structure to be a homodimer (beta-alpha-alpha-beta) similar to human Hp 1-1. Interestingly, Western blot and high performance liquid chromatographic (HPLC) analysis showed that 25% of the TLY minipigs possessed low or no plasma Hp level (<0.05 mg/ml). The Hp cDNA of these TLY minipigs was then cloned, and the translated amino acid sequence was analyzed. No sequences were found to be deficient; they showed a 99.7% identity with domestic pigs (NP_999165). The mean overall Hp level of the TLY minipigs (0.21 +/- 0.25 mg/ml; n=43) determined by enzyme-linked immunosorbent assay (ELISA) was markedly lower than that of domestic pigs (0.78 +/- 0.45 mg/ml; p<0.001), while 25% of the TLY minipigs had an Hp level that was extremely low (<0.05 mg/ml). In addition, the initial recovery rate (first 40 min) in the circulation of infused fluorescein isothiocyanate (FITC)-Hb was significantly higher in the TLY minipigs with extremely low Hp levels than those with high levels. This data suggests that the low concentration of Hp-Hb complex is responsible for the higher recovery rate of Hb in the circulation. TLY minipigs have been used as an experimental model for cardiovascular diseases; whether they can be used as a model for inflammatory diseases, with Hp as a marker, remains a topic of interest. However, since the Hp level varies significantly among individual TLY minipigs, it is necessary to prescreen the Hp levels of the animals to minimize variation in the experimental baseline. The present study may provide a reference value for future use of the TLY minipig as an animal model for inflammation-associated diseases.