Stimulation of 2',5'-oligoadenylate synthetase activity in sheep endometrium during pregnancy, by intrauterine infusion of ovine trophoblast protein-1, and by intramuscular administration of recombinant bovine interferon-alpha I1.

In Expt 1, activity of 2',5'-oligoadenylate (2',5'-A) synthetase in endometrium collected on Day 16 (oestrus is Day 0) from the uterine horn ipsilateral to the corpus luteum was greater (P less than 0.001) for pregnant (135.5 +/- 1.72 nmol/mg protein/h) than for cyclic ewes (58.5 +/- 0.99 nmol/mg protein/h). In pregnant ewes, activity of 2',5'-A synthetase in endometrium collected from the contralateral uterine horn (119.5 +/- 1.72 nmol/mg protein/h) did not differ from that of the ipsilateral horn. In Expt 2, three ovariectomized ewes were treated with progesterone for 10 days and then with oestrogen for 2 days. Activity of 2',5'-A synthetase on Day 13 was 18% greater (P less than 0.10) in endometrium collected from the uterine horn receiving infusions of 30 micrograms ovine trophoblast protein-1 (oTP-1) twice a day on Days 10, 11 and 12(57.7 +/- 0.22 nmol/mg protein/h) than from the uterine horn receiving control infusions of serum protein (SP; 48.8 +/- 0.22 nmol/mg protein/h). In Expt 3, activity of 2',5'-A synthetase on Day 15 was not significantly greater in endometrium collected from the uterine horn of cyclic ewes receiving infusions of 30 micrograms oTP-1 twice a day on Days 12, 13 and 14 (46.5 +/- 0.37 nmol/mg protein/h) than in endometrium from the uterine horn receiving infusions of SP (38.2 +/- 0.37 nmol/mg protein/h). When results of Expt 2 and Expt 3 were combined, intrauterine infusion of oTP-1 increased (P less than 0.05) activity of 2',5'-A synthetase in endometrium by 20%.(ABSTRACT TRUNCATED AT 250 WORDS)     

Interrelationships of contraction time, Vmax, and myosin ATPase after spinal transection

Interrelationships of selected mechanical and biochemical properties of hindlimb extensor muscles following low thoracic cord transection were studied. Kittens were spinalized (Sp) at 2 wk and maintained for 6-12 mo. Some Sp animals were exercised (Sp-E) on a treadmill 25-30 min/day, 5 days/wk. In situ contractile properties of the slow-twitch soleus (SOL) and fast-twitch medical gastrocnemius (MG) muscles of normal (N), Sp, and Sp-E cats were determined. Exercise did not affect most parameters; thus Sp and Sp-E groups are considered collectively. The cross-sectional areas (CSA) of the SOL and MG decreased by 43 and 32%, respectively. Specific tension (tension/CSA) was maintained in the SOL but decreased (P less than 0.05) in the MG. Contraction time (CT) and half-relaxation time were significantly shorter in the SOL but unchanged in the MG. Maximum shortening velocity (Vmax) and myosin ATPase (mumol X mg-1 X min-1) increased (P less than 0.05) in the SOL of both groups and the MG of Sp. Frequency-tension responses of both muscles shifted toward that resembling a "faster" muscle. These results substantiate the existence of relatively independent regulatory mechanisms for Vmax and CT and show that myosin ATPase levels are more closely related to Vmax than CT. Although the changes in the SOL were consistent with the hypothesis that slow fibers are converted to fast, the elevated Vmax and myosin ATPase of the MG suggest that significant changes also occur within a "fast" fiber-type category.     

Comparison of hepatic drug metabolizing enzyme activities in several agricultural species

1. Several pathways of drug metabolizing enzyme activity were measured in hepatic fractions of cattle, sheep, goats, chickens, turkeys, ducks, rabbits and rats. The pathways examined included the O-demethylation of p-nitrophenol, microsomal ester hydrolysis of procaine and glucuronidation of p-nitrophenol, and the cytosolic acetylation of sulfamethazine and sulfation of 2-naphthol. 2. For most enzymatic pathways measured, goats were more similar to sheep (wether) than to cattle (steers). The exception was UDP-glucuronyltransferase activity, which was significantly higher for the goat than for any other species studied. 3. Within the avian subset, the chicken and turkey were usually the most similar species. 4. The activities of arylsulfotransferase isozymes III and IV were particularly low for the duck compared to the chicken and turkey. 5. N-acetyltransferase activity was very high for rabbits and very low for sheep and goats.     

Comparison of lipid/gramicidin dispersions and cocrystals by Raman scattering

Gramicidin crystals, dimyristoylphosphatidylcholine (DMPC)/gramicidin dispersions, and DMPC/gramicidin cocrystals were examined by Raman scattering to determine lipid/gramicidin stoichiometries and lipid organization. Calibrations of the choline (716-cm-1) and tryptophan (756-cm-1) peaks indicate that the cocrystals contain two lipids for each gramicidin monomer, a result confirmed by chemical analyses of washed crystals. In dispersions with high lipid/gramicidin ratios (e.g., 25:1), the lipid is ordered but becomes increasingly disordered as the gramicidin content is increased. Paradoxically, the DMPC/gramicidin cocrystals have highly ordered lipids that possibly contain no gauche bonds at all, despite their low lipid/gramicidin ratio. In addition, the polypeptide amide I peak position near 1670 cm-1 is found to be independent of the lipid/gramicidin ratio in the complexes and may indicate a beta-helix-type secondary structure at all ratios. However, the amide I peak broadens significantly at low lipid/gramicidin ratios and broadens still further in the cocrystals, suggesting that protein-protein interactions may induce band-broadening distortions of the polypeptide structure.     

Survival and behaviour of unfed stages of the ticksRhipicephalus appendiculatus,Boophilus decoloratus andB. microplus under field conditions in Zimbabwe

The survival and behaviour of the unfed stages ofRhipicephalus appendiculatus, Boophilus decoloratus andB. microplus in gauze columns were observed in long and short grass in the highveld of Zimbabwe. Ticks were exposed in the cool, hot and rainy seasons of 1980 and 1981. All species and stages survived longer in long grass than in short grass. Larvae from engorged female ticks released in the cool season hatched much later than incubator-reared controls. They were consequently not present during the cold weather and survived longer than larvae subjected to the low temperatures, in which the shortest survival-times were recorded. The survival of nymphs was insensitive to season. The longest survival-times were recorded in adults. Median survival-times of incubator-reared adults ranged from 165 to 375 days in short grass and from 333 to 493 days in long grass. These times were usually longer than those for adults which moulted in the field. Larvae of the three species and nymphs ofR. appendiculatus were active soon after hatching or moulting, irrespective of the season. In contrast, adults ofR. appendiculatus showed different patterns of activity in different seasons. Adults first appeared at the base of the columns in October/November and then gradually ascended to reach a maximum height in December/January. They remained high up in the columns until May/June when the weather became increasingly cold and dry. Larvae ofB. decoloratus climbed higher up in the columns in the long grass than did the larvae of the other two species.Larvae and nymphs ofR. appendiculatus and larvae ofB. microplus migrated up and down the columns daily, but larvae ofB. decoloratus and adults ofR. appendiculatus did not migrate.     

Global proteome discovery using an online three-dimensional LC-MS/MS

We have developed a proteomics technology featuring on-line three-dimensional liquid chromatography coupled to tandem mass spectrometry (3D LC-MS/MS). Using 3D LC-MS/MS, the yeast-soluble, urea-solubilized peripheral membrane and SDS-solubilized membrane protein samples collectively yielded 3019 unique yeast protein identifications with an average of 5.5 peptides per protein from the 6300-gene Saccharomyces Genome Database searched with SEQUEST. A single run of the urea-solubilized sample yielded 2255 unique protein identifications, suggesting high peak capacity and resolving power of 3D LC-MS/MS. After precipitation of SDS from the digested membrane protein sample, 3D LC-MS/MS allowed the analysis of membrane proteins. Among 1221 proteins containing two or more predicted transmembrane domains, 495 such proteins were identified. The improved yeast proteome data allowed the mapping of many metabolic pathways and functional categories. The 3D LC-MS/MS technology provides a suitable tool for global proteome discovery.     

Firefly luciferase as a reporter of regulated gene expression in higher plants

The firefly luciferase, assayedin vivo with a low-light video camera, acts as a non-invasive, real-time reporter of the temporal and spatial regulation of gene expression in single plants. Furthermore, the sensitivity of the luciferase assay in extracts of transformed plant tissue makes it a particularly useful marker in transient or stable transformation experiments.