Sequence analysis of leukocyte chemoattractant peptides secreted by periovulatory ovine follicles.

Cells of the white blood series infiltrate ovarian follicles during the ovulatory process. A segment of the wall of periovulatory ovine follicles was incubated and conditioned media subjected to ultrafiltration. Leukocyte chemoattractant activity of media was measured using a linear under-agarose migration assay. Bioactivity was recovered following filtration through a 3000 molecular weight cut-off membrane. Filtrate was then fractionated by reverse phase high performance liquid chromatography. Peptides recovered from two fractions with significant chemoattractant activity were sequenced. One fraction contained 16 amino acid residues with repeating triplets of Gly-X-Y, where X and Y were often proline and hydroxyproline, respectively. Because this motif is characteristic of alpha collagens, and since thecal collagen is degraded during the mechanics of ovulation, it appears that this chemoattractant is derived from the connective tissue matrix of the follicle. Peptide isolated from the other bioactive chromatographic fraction was 15 amino acids in length, and rich in glycine, but did not contain imino acids. To our knowledge this is the first report of purification of leukocyte chemoattractants of reproductive tissue origin. Resident follicular granulocytes and mononuclear cells are capable of secreting a broad spectrum of potent chemicals that could be involved in the mechanisms of ovulation and luteinization.     

Streamflow-induced variations in nitrate flux in tributaries to the Atlantic Coastal Zone

Streamflow-related variability in nutrient flux represents an important source of uncertainty in managing nutrient inputs to coastal ecosystems. Quantification of flux variability is of particular interest to coastal resource managers in adopting effective nutrient-reduction goals and monitoring progress towards these goals. We used historical records of streamflow and water-quality measurements for 104 river monitoring stations in an analysis of variability in annual and seasonal flux of nitrate to the Atlantic coastal zone. We present two measures of temporal flux variability: the coefficient of variation (CV) and the exceedence probability (EP) of 1.5 times the median flux. The magnitude of flux variations spans a very wide range and depends importantly upon the season of year and the climatic and land-use characteristics of the tributary watersheds. Year-to-year variations (CV) in annual mean flux range over two orders of magnitude, from 3–200% of the long-term mean flux, although variations more typically range from 20–40% of the long-term mean. The annual probability of exceeding the long-term median flux by more than 50% (EP) is less than 0.10 in most rivers, but is between 0.10 and 0.35 in 40% of the rivers. Year-to-year variability in seasonal mean flux commonly exceeds that in annual flux by a factor of 1.5 to 4. In western Gulf of Mexico coastal rivers, the year-to-year variablity in the seasonal mean flux is larger than in other regions, and is of a similar magnitude in all seasons. By contrast, in Atlantic coastal rivers, the winter and spring seasons, which account for about 70% of the annual flux, display the smallest relative variability in seasonal mean flux. We quantify the elasticity of nutrient flux to hypothetical changes in Streamflow (i.e., the percent increase in flux per percentage increase in mean discharge) to allow the approximation of flux variability from streamflow records and the estimation of the effects of future climatically-induced changes in Streamflow on nutrient flux. Flux elasticities are less than unity (median = 0.93%) at most stations, but vary widely from 0.05% to 1.59%. Elasticities above unity occur most frequently in the largest rivers and in rivers draining the arid portions of the western Gulf of Mexico Basin. Historical flux variability and elasticity generally increase with the extent of arid conditions and the quantity of nonurban land use in the watershed. We extend the analysis of flux variability to examine several case studies of highly unusual meteorological events capable of significantly elevating nitrate flux and degrading estuarine ecology.     

Production of low molecular weight chemoattractants for leukocytes by periovulatory ovine follicles

Studies were performed to determine if periovulatory ovine follicles secrete chemoattractants for leukocytes, and if so, to begin to elucidate the chemical nature of such factors. Tissues were obtained at 0, 12, 24, and 36 h after initiation of the preovulatory surge of luteinizing hormone and placed in short-term incubation (ovulation occurs at approximately 24 h). Follicular-conditioned medium was tested for its ability to attract leukocytes by utilizing a linear under-agarose assay: chemotaxis was quantified as a function of the leading front of migration of cells. Neutrophils and eosinophils were attracted toward media conditioned with tissues of 24 and 36 h. Monocytes responded toward medium of tissues collected at 36 h. There was no evidence for chemoattraction of basophils or lymphocytes. Chemoattractant activity for granulocytes and monocytes was of low molecular weight origin (less than 3000) and water-soluble. High-performance liquid chromatographic separation of this sample produced a distinct peak with recoverable activity. The isolated fraction was rich in glycine. Eosinophils also migrated toward an additional low molecular weight attractant that was extracted into ethyl acetate. Leukocytes attracted into periovulatory follicles might produce substances (eg., proteolytic enzymes and angiogenic factors) that play a role in the mechanisms of ovulation and luteinization.     

Clinical Trial of Mefruside,a New Diuretic

A controlled clinical trial of a new diuretic—mefruside—is reported, in which it was compared with frusemide in 15 normal subjects and 15 patients with fluid retention. It was found to be an effective diuretic which, in the patients, produced a significantly greater excretion of water and electrolytes than an equal dose of frusemide. Its smooth prolonged action, maximal in the first 12 hours, made it of particular value for maintenance therapy. In a short-term trial on a further 15 hypertensive patients mefruside was shown to have a useful hypotensive action. The drug was well tolerated with minimal side effects.