Sex differences in reinforcing, analgesic and other effects of opioids have been demonstrated; however, the extent to which sex differences in motoric effects of opioids contribute to apparent sex differences in their primary effects is not known. The goal of this study was to compare the effects of the prototypic mu opioid agonist morphine on locomotor activity in male vs. female rats. Saline or morphine (1-10 mg/kg) was administered s.c. to adult Sprague-Dawley rats, which were placed into a photobeam apparatus for 3-5 h to measure activity. Modulation of morphine's effects by gonadal hormones and by handling (either during the test session or for 4 days before the test session) were examined. Morphine initially suppressed and later increased locomotor activity in both sexes relative to their saline-injected controls, but males were more sensitive than females to the initial locomotor suppressant effect of morphine. Intermittent, brief handling during the 3-h test session blunted morphine-induced locomotor activation in both sexes. Females in proestrus were the most sensitive to morphine's locomotor-stimulant effect, with females in estrus showing the least response to morphine. Gonadectomized (GDX) males with or without testosterone were equally sensitive to morphine's effects, whereas GDX females treated with estradiol showed a blunted response to morphine's effects, similar to intact females in estrus. Brief handling on each of 4 consecutive days pre-test attenuated morphine's locomotor suppressant effect in males but had no effect in females, thereby eliminating the sex difference. These data suggest that sex differences in morphine's effects on locomotor activity can be attributed to gonadal hormones in females, and to differential stress-induced modulation of morphine's effects in males vs. females.

Significant increases in the production of winter cereals could be realized with improved hardiness. The objective of this research was to determine if overwintering cereals modify water soluble carbohydrate composition in response to naturally occurring winter freeze stress. Fields of wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), and rye (Secale cereale L.) were monitored throughout the winter between 1986 to 1990 for snow-pattern development, temperature, and snowfall. Plant crowns were analyzed for freeze injury (form and intensity) and water soluble carbohydrate composition (froctan, sucrose, glucose, and fnlctose). Growth chamber plants were frozen and analyzed to confirm the interpretation of field plant data. In 1986, there was less froctan and more soluble sugars in the exposed than in the snow-covered wheat plants (but no complete depletion of fructan), and no crown injury from equilibrium freezing. In 1987· 1988, little injury or froctan hydrolysis occurred in barley, wheat, or rye; the plants were snow covered during cold periods. In -1990, fructan hydrolysis to sugar in exposed barley, wheat, and rye plants paralleled that in the exposed wheat plants of [1986][1987]. The fructan content of exposed barley plants was nearly depleted during the coldest weather. Exposed plants had typical equilibrium freeze injury. Growth chamber plants responded similarly to the exposed field plants. Plants exposed to freeze stress converted fructan to sugars which probably alleviated adhesive freeze stress. The energy of hydrolysis may be useful for placing sugar into sites within the tissue where its cryoprotective activity is most effective.

Marek's disease herpesvirus A antigen was purified greater than 200-fold with a 24% recovery by ion exchange column chromatography, isoelectric focusing, and preparative polyacrylamide gel electrophoresis. The antigen had an isoelectric point of 6.68 i 0.03 in the presence of 1 M urea and 0.05% Brij 35, a nonionic detergent, and approximately 6.5 in the absence of dissociating agents. When analyzed by electrophoresis on analytical polyacrylamide gels, the purified antigen migrated as a single broad band which stained for both protein and carbohydrate, suggesting that it was a highly purified heterogeneous glycoprotein. However, the antigen was not purified to homogeneity as determined by electrophoresis on polyacrylamide gels in the presence of sodium dodecyl sulfate and by immunodiffusion analysis. Antibody to Marek's disease herpesvirus A antigen was prepared in a rabbit, and antibody to two contaminating antigens was removed by adsorption to yield monospecific antisera. A method for large scale production of Marek's disease herpesvirus A antigen (MDHV-A antigen) yields sufficient antigen to facilitate its purification and physical and chemical characterization (8). Initial characterization of unpurified or partially purified antigen in this laboratory (8) and by others (11, 13, 14) revealed that MDHV-A antigen is a glycoprotein stable at pH 2. The antigen sediments at about 3.7S (8) to 4.2S (11), its apparent molecular weight is estimated to range from 33,000 (11) to 44,850 (8) and 80,000 (13), and its isoelectric point ranges from 4.5 to 6.35 (11, 13). Because of these variations (8, 11, 13), it became clear that the antigen would have to be purified more extensively to obtain precise estimates of physical parameters, as well as to determine its biological properties. Initial purification ranges from

Sera from chickens naturally infected with Marek's disease herpesvirus (MDHV) form precipitin lines with at least two immunologically distinct soluble antigens designated MDHV-A and MDHV-B. Partial purification and characterization of the glycoprotein MDHV-A antigen was previously reported. MDHV-B was found predominantly in the sonically treated extracts of infected cells, in contrast to the predominantly extracellular MDHV-A. Analysis of these extracts from [14C]glucosamine-labeled cells by immunodiffusion with chicken anti-MDHV-B serum negative for MDHV-A followed by autoradiography confirmed that MDHV-B was a common antigen between MDHV and herpesvirus of turkeys and revealed that it was also a glycoprotein. Because of their glycoprotein nature, both MDHV-A and MDHV-B bound to concanavalin A affinity chromatography columns and could then be eluted by a-methyl-D-mannoside and recovered for further analysis. Concanavalin A affinity chromatography was an excellent technique for initial purification of MDHV-A and MDHV-B, since approximately 5and 15-fold purification, respectively, was achieved in a single simple step. MDHV-B was resistant to trypsin under conditions where MDHV-A was sensitive, but was similar to MDHV-A in resistance to pH 2.0 and to 1.0 or 2.0 M urea and 0.05% Brij 35. Partially purified MDHV-B was analyzed by sucrose gradient sedimentation, isoelectric focusing, and gel filtration on Sephadex G-200 in the presence of 1.0 or 2.0 M urea and 0.05% Brij 35 to purify the antigen and to determine its physical and chemical properties in comparison with those already reported for MDHV-A. MDHV-B had a much lower isoelectric point in pH 4.54, a higher sedimentation coefficient of 4.4S, and a greater molecular weight of 58,250. These data indicate that MDHV-B is physically distinct from MDHV-A antigen, although the size difference is not sufficient to allow for effective separation. In contrast, the isoelectric point difference of greater than 2 pH units makes isoelectric focusing an effective means of purifying the antigens free of one another. The four-step purification procedure achieved greater than 200-fold purification of MDHV-B. Immunization of rabbits with this highly purified antigen results in the preparation of antisera that appeared monospecific for MDHV-B in immunodiffusion. Three antigens are detected most consistently by immunodiffusion analysis of Marek's disease herpesvirus (MDHV)-infected cell extracts and concentrated culture fluid from infected cells using sera from naturally infected chickens (3, 4). They include "A antigen," found in both the culture fluid and the extracts of infected cells, and "B and C antigens," found primarily in the extracts of infected cells. In addition, Marek's disease (MD)-related membrane antigen (1, 2, 15) is detected on the surface of MDHV-infected cells by immunofluorescence. MD membrane

Deficiencies in winter hardiness of barley (Hordeum vulgare L.) with respect to osmotic and adhesive components of freeze stress might be improved by genetic transfer of specific traits from rye (Secale cereale L.). These protective traits may include the hydrolysis of fructan for accumulation and secretion of sugars. The objectives of this research were (i) to measure total and intercellular water soluble carbohydrates (CLIO), sucrose, glucose, fructose, and fructan, in ‘Hudson’ barley and ‘Rosen’ rye by ethanol/water extraction of plants and by perfusion of crowns; and (ii) to estimate relief of freeze stress caused by adhesive and osmotic energies. Carbohydrates were detected and quantified by refractometry after high pressure liquid chromatography (HPLC). Freezing (−3°C, 4 d) caused a greater change the molar concentration of CHOs in rye than in barley extracts (26 vs. 16% increase, respectively), which would proportionally decrease freeze dehydration. The sugars perfused from frozen plants consisted of a higher proportion of monosaccharides in rye than barley, which would result in a greater potential for relief of adhesion. These mechanisms involve increase of sugar content in locations which relieve critical freeze stress.