Steroidogenesis during induced oocyte maturation and ovulation in the African catfish,Clarias gariepinus

Changes in ovarian steroidogenesis accompanying oocyte maturation and ovulation were studied in the African catfish,Clarias gariepinus. Laboratory-reared females with postvitellogenic ovaries were treated with pimozide and LHRH-analogue. The plasma gonadotropin levels were determined by means of a homologous radioimmunoassay, the condition of the ovaries was studied by histological examination of the follicles, and the steroidogenetic capacity of the ovaries was analyzed byin vitro incubation of tissue fragments for 3 h with [³H]-pregnenolone and [³H]androstenedione as precursors. Data were collected at regular intervals between 0 and 16 h after pimozide-LHRH analogue administration.Until 4 h after the beginning of the experiments the plasma gonadotropin levels did not rise above 25 ng/ml, the ovaries remained in the stage of postvitellogenesis, and testosterone was the main end product of steroidogenesis. Four hours later the gonadotropin concentration in the blood had risen to more than 150 ng/ml, and the ovaries had entered the stage of germinal vesicle migration. At the same time steroidogenesis shifted towards the production of 17,20-dihydroxy-4-pregnen-3-one, 5-pregnane-3, 17-diol-20-one, 5-pregnane-3,6,17-triol-20-one, 5-pregnane-3,17,20-triol and 5-pregnane-3,6,17,20-tetrol. During the subsequent stage of germinal vesicle breakdown the plasma gonadotropin level remained high, and the synthesis of the C₂₁-steroids showed a further increase. Simultaneously, the production of some C₁₉-steroid glucuronides was enhanced. The preovulation and especially the postovulation stages were accompanied by a gradual decrease in steroidogenic capacity of the ovaries, even though the plasma gonadotropin level remained high. It is concluded that the prematuration surge of gonadotropin influences the activity of enzymes involved in steroidogenesis, leading to a reduced C_17,20-lyase and to an augmented activity of the enzymes 20-hydroxysteroid dehydrogenase (HSD), 5-reductase, 3-HSD, 6-hydroxylase and UDP-glucuronosyltransferase. During ovulation the activity of all steroidogenic enzymes, including such key enzymes as 3-HSD and 17-hydroxylase, gradually decreases.Not only 17,20-dihydroxy-4-pregnen-3-one, but also the 5-reduced pregnanes may be involved in inducing oocyte maturation and/or ovulation. The very polar triol and tetrol products may function, together with the steroid glucuronides as sex pheromones.A preliminary account of these results was presented at the XIII Conference of European Comparative Endocrinologists, Belgrade, September 7–12, 1986     

Mainestay: A high-yielding, round white potato variety for fresh market

The Mainestay potato variety is high-yielding and late-maturing producing attractive, round, white-skinned, white-fleshed tubers with medium-shallow eyes. Its major use is expected to be as a maincrop tablestock variety. Mainestay appearance ratings are similar to those of Katahdin, and greening in Mainestay tubers is less than in Kennebec. Most boil and bake qualities were rated equal to Katahdin in at least half of the tests, and Mainestay usually has a low incidence of internal defects. However, purple streak, heat necrosis, vascular discoloration, and heat sprouts have been seen on occasion. Mainestay tubers do not show the net necrosis caused by potato leafroll virus, and have shown very little hollow heart or blackspot bruising. Mainestay is susceptible to common scab and acid scab, but moderately resistant to verticillium wilt (Verticillium, albo-atrum andV. dahliae), early blight (Alternaria solani), rhizoctonia (Rhizoctonia solani), dry rot (Fusarium sambucinum), and pink rot (Phytophthora erythroseptica). It is moderately susceptible to the common race of late blight (Phytophthora infestans). Mainestay tubers are slow to mature after vine kill, and additional time is required prior to harvest in order to allow for proper periderm development and to reduce storage losses.     

Phenotypic stability of resistance to late blight in potato clones evaluated at eight sites in the United States

Changes in the fungal pathogenPhytophthora infestans in the United States pose a significant threat to potato production. Sources of resistance to these new genotypes of P.infestans need to be identified for potato breeders to have parental materials for crossing, and the phenotypic stability of late blight resistance in these potato clones needs to be determined. Sixteen potato clones which reportedly have some resistance to late blight were evaluated at eight locations: Florida (FL), Maine (ME), Michigan (MI), Minnesota (MN), North Dakota (ND), New York (NY), Pennsylvania (PA) and Wisconsin (WI) in 1996. Percent infected foliage was recorded at approximately weekly intervals following the onset of the disease at each location. Area under the disease progress curve (AUDPC) was calculated. Clones were ranked for mean AUDPC within location and the nonparametric stability statistics, mean absolute rank differences and variance of the ranks, were analyzed for phenotypic stability. Neither of these statistics was significant, indicating a lack of genotype x environment interaction on the rankings of these clones across locations in 1996. The four clones with lowest AUDPC scores were U.S. clones AWN86514-2, B0692-4, B0718-3 and B0767-2. These clones should be useful parental materials for breeders seeking to incorporate genes for late blight resistance into potatoes.     

A 13CO2 Enrichment Experiment to Study the Synthesis Pathways of Polyunsaturated Fatty Acids of the Haptophyte Tisochrysis lutea

The production of polyunsaturated fatty acids (PUFA) in Tisochrysis lutea was studied using the gradual incorporation of a ¹³C-enriched isotopic marker, ¹³CO₂, for 24 h during the exponential growth of the algae. The ¹³C enrichment of eleven fatty acids was followed to understand the synthetic pathways the most likely to form the essential polyunsaturated fatty acids 20:5n-3 (EPA) and 22:6n-3 (DHA) in T. lutea. The fatty acids 16:0, 18:1n-9 + 18:3n-3, 18:2n-6, and 22:5n-6 were the most enriched in ¹³C. On the contrary, 18:4n-3 and 18:5n-3 were the least enriched in ¹³C after long chain polyunsaturated fatty acids such as 20:5n-3 or 22:5n-3. The algae appeared to use different routes in parallel to form its polyunsaturated fatty acids. The use of the PKS pathway was hypothesized for polyunsaturated fatty acids with n-6 configuration (such as 22:5n-6) but might also exist for n-3 PUFA (especially 20:5n-3). With regard to the conventional n-3 PUFA pathway, Δ6 desaturation of 18:3n-3 appeared to be the most limiting step for T. lutea, “stopping” at the synthesis of 18:4n-3 and 18:5n-3. These two fatty acids were hypothesized to not undergo any further reaction of elongation and desaturation after being formed and were therefore considered “end-products”. To circumvent this limiting synthetic route, Tisochrysis lutea seemed to have developed an alternative route via Δ8 desaturation to produce longer chain fatty acids such as 20:5n-3 and 22:5n-3. 22:6n-3 presented a lower enrichment and appeared to be produced by a combination of different pathways: the conventional n-3 PUFA pathway by desaturation of 22:5n-3, the alternative route of ω-3 desaturase using 22:5n-6 as precursor, and possibly the PKS pathway. In this study, PKS synthesis looked particularly effective for producing long chain polyunsaturated fatty acids. The rate of enrichment of these compounds hypothetically synthesized by PKS is remarkably fast, making undetectable the ¹³C incorporation into their precursors. Finally, we identified a protein cluster gathering PKS sequences of proteins that are hypothesized allowing n-3 PUFA synthesis.     

Application of a PCR-based test for detection of potato late blight and pink rot in tubers

A polymerase chain reaction (PCR)-based test for potato late blight (Phytophthora infestans) and pink rot (P.erythroseptica, P. nicotianae) diseases has been developed for use with potato tuber tissue. Primers based on sequence analysis of the ITS2 region of ribosomal DNA of late blight and pink rot pathogens were utilized in PCR assays of inoculated tubers and tubers harvested from plots known to have late blight and/or pink rot. Assays of artificially inoculated Kennebec and Russet Burbank tubers revealed thatP. infestans was detected by PCR as early as 72 h after inoculation and in the absence of visible symptoms. Much higher detection frequencies were obtained by PCR compared with plating on selective medium or placement of tissue in moist chambers. Tubers from plots known to have late blight and/or pink rot were tested using the PCR assay. Assay of late blight lesions showed ca. 80% recovery for late blight-infected tubers from the field. Results indicate that the PCR assay provides a rapid and accurate test for diagnosis of late blight and pink rot in potato tubers.     

Effects of Land‐Related Policies on Land Development during a Real Estate Boom and a Recession

Our research evaluates and compares the effects of three land‐related policies on land development in a sprawling metropolitan area during a real estate boom and a recession. Our findings suggest that during a real estate boom 1) the urban growth boundary (UGB) serves its purpose of attracting urban development inside the given boundary during a boom while its effectiveness diminishes with increased development pressure from lower‐valued land outside of the UGB during a recession period, 2) the agricultural zone works well for restraining new development during a boom period while agriculture zoned parcels are more likely developed during a recession period, and 3) an increase in the land‐value tax bill increases the incentive for development during a recession period while higher tax bills do not affect development during a boom period. In anticipation of a re‐emergence of urban sprawl with the recovery of the real estate market, our findings imply that land planners and others concerned with sprawling development should pay more attention to development in sprawl‐prone areas during recession periods.     

Atomic structure of PDE4: insights into phosphodiesterase mechanism and specificity

Cyclic nucleotides are second messengers that are essential in vision, muscle contraction, neurotransmission, exocytosis, cell growth, and differentiation. These molecules are degraded by a family of enzymes known as phosphodiesterases, which serve a critical function by regulating the intracellular concentration of cyclic nucleotides. We have determined the three-dimensional structure of the catalytic domain of phosphodiesterase 4B2B to 1.77 angstrom resolution. The active site has been identified and contains a cluster of two metal atoms. The structure suggests the mechanism of action and basis for specificity and will provide a framework for structure-assisted drug design for members of the phosphodiesterase family.     

Effects of dietary ruminally protected L-carnitine on plasma metabolites in sheep following a sub-lethal ammonia challenge

In Experiment 1, lambs were randomly assigned to 0.25, 1.00, 2.50, 5.00 and 10.00 g/day of dietary ruminally protected L-carnitine (RPLC) and were allowed to adapt for 20 days. Plasma samples were obtained at 0, 120 and 240 min after RPLC feeding. Plasma L-carnitine (LC) concentrations increased (p<0.01) for all levels of RPLC treatment, however, no differences were observed due to level of RPLC or time. Plasma LC concentrations were 27.05 and 57.83 micromol/l for baseline and pooled RPLC treated sheep, respectively. In Experiment 2, lambs were randomly assigned to 0, 0.125, 1.06 and 2.0 g/day of RPLC and were adapted as in Experiment 1. Plasma was collected at 0, 15, 30, 60, 90, 180, 240 and 360 min after oral ammonia challenge (300 mg/kg BW urea). Plasma LC concentrations increased with treatment relative to control (p<0.01). Plasma LC concentrations were 35.7, 44.2, 60.5 and 65.7 micromol/l for the 0, 0.125, 1.06 and 2.0 g/day treatments, respectively. RPLC tended to decrease plasma ammonia at some time points (time x treatment; p=0.10). We conclude that RPLC increased plasma LC concentrations, but had only modest effects on plasma ammonia concentrations and had no effect on plasma urea or glucose concentrations.