The effect of octadecenoic acid on the growth of japanese encephalitis virus in Novikoff hepatoma cells.

The growth characteristics of Japanese encephalitis virus cultivated in Novikoff hepatoma cells grown in shaker culture can be differentially altered by the presence of 6-cis or cis-9-octadecenoic acid in Swim's 67-G medium. The addition of 125 mug of the 6-isomer per ml medium reduced the number of infectious particles produced, whereas the same amount of the 9-isomer enhanced virus production. The virus was found to be more stable in cell-free spent medium than in fresh medium. The presence of 125 mug 6-18:1 per ml in fresh medium resulted in a rapid loss of virus infectivity.     

Amenorrhoea after Discontinuing Combined Oestrogen-Progestogen Oral Contraceptives

Out of 210 women seen at the Middlesex Hospital with secondary amenorrhoea the 63 who developed it after stopping oral contraceptives were fully investigated. Five had organic disease sufficient to account for the amenorrhoea (one had severe diabetes, one a pituitary tumour, and three premature ovarian failure); two patients had galactorrhoea (one of whom also had the pituitary tumour); two had anorexia nervosa.Of the 63 women 40 (63%) had suffered from amenorrhoea or prolonged or irregular menstrual cycles before taking the pill, and this suggested that combined oestrogen-progestogen oral contraceptives should be used with caution for women with irregular menstruation.Nineteen patients wished to become pregnant and 12 have so far done so after treatment with clomiphene or gonadotrophins.In another study 204 women recorded when their first menstrual cycle occurred after stopping the pill. Seventy-four had a cycle longer than five weeks but only five exceeded three months, and only one of the five had more than six months'' amenorrhoea. These results confirm that the incidence of amenorrhoea after stopping oral contraceptives is low.     

Some effects of tentoxin on mature and developing chloroplasts

Tentoxin affected chloroplasts of all stages of development; it caused chlorosis of expanded and developing bean and lettuce leaves when introduced through the stem or roots and inhibited greening of etiolated lettuce seedlings. Cotyledons from 6-day-old lettuce seedlings grown in 10ug/ml tentoxin bad 50–68% of the monogalactosyldiglyceride and digalactosyldiglyceride content of healthy tissue whereas the sulfolipid, phospatidyl-glycerol, -ethanolamine, -choline and -inosrtol levels were unchanged. Toxin-treated tissue also showed selective increases in C-16 and C-18 saturated fatty acids.Research cooperative with the College of Agricultural and Life Sciences, University of Wisconsin, Madison, and the Agricultural Research Service, U.S. Department of Agriculture.Mention of companies or commercial products does not imply recommendation or endorsement by the U.S. Department of Agriculture over others not mentioned.     

Morphology and growth kinetics of hyphae of differentiated and undifferentiated mycelia of Neurospora crassa.

A comparison was made of the morphology and growth kinetics of hyphae of differentiated and undifferentiated mycelia of Neurospora crassa. Undifferentiated mycelia were formed during exponential growth on solid media or submerged culture. Hyphae at the margin of differentiated mycelia (colonies) differed from undifferentiated mycelia in diameter, extension rate, extension zone length, and intercalary and apical compartment length. The mean hyphal extension rate (E) of an undifferentiated mycelium was a function of the length of the mycelium's hyphal growth unit (G) and the organism's specific growth rate (alpha). Thus, E=Galpha.     

Regulation of Oleoresinosis in Grand Fir (Abies grandis) (Coordinate Induction of Monoterpene and Diterpene Cyclases and Two Cytochrome P450-Dependent Diterpenoid Hydroxylases by Stem Wounding)

Oleoresin (pitch) is a defensive secretion composed of monoterpene olefins (turpentine) and diterpene resin acids (rosin) that is produced in grand fir (Abies grandis Lindl.) stems in response to wounding. Monoterpene and diterpene biosynthesis are coordinately induced in wounded stems as determined by monitoring the activity of monoterpene and diterpene cyclases, as well as two cytochrome P450-dependent diterpenoid hydroxylases involved in the formation of ([mdash])-abietic acid, the principal resin acid of this species. The activity of these enzymes reaches maximum levels that are 5- to 100-fold higher than those of nowwounded control stems 10 d after wounding and this is followed by a synchronous decline. The increase in biosynthetic activity is consequently followed by the accumulation of a viscous mass of resin acids, with the loss of the volatile monoterpenes, at the site of injury. The observed coordinate induction of monoterpene olefin and abietic acid bio-synthesis and the results of oleoresin analysis are consistent with the role of the volatile monoterpenes as a solvent for the mobilization and deposition of resin acids at the wound site to seal the injury with a rosin barrier after the evaporation of the turpentine. The last step of resin acid biosynthesis is catalyzed by an operationally soluble aldehyde dehydrogenase that is not inducible by wounding but seemingly is expressed constitutively at a high level. In vivo [14C]acetate feeding and resin analysis indicate that this enzyme is not efficiently coupled to the earlier steps of the pathway.     

Growth and disinfestation of 6 different bacteria in embryogenic suspension cultures of cotton

Summary Growth of 6 different common laboratory bacteria (Escherichia coli, Flavobacterium balustrum, Xanthomonas maltophilia, Enterobacter cloacae, Pseudomonas fluorescens, and Agrobacterium tumefaciens) in a bacterial medium, fresh plant medium, and spent plant media was initially measured. In all cases, bacteria grew best in the bacterial medium followed by the fresh plant medium. The spent plant medium did not support growth of the bacteria and apparently was actively toxic to bacterial cells. Proliferating, embryogenic suspension cultures of cotton (Gossypium hirsutum) were then inoculated with these 6 different bacteria. Two to three d following bacterial inoculation, embryogenic tissues were placed in various concentrations of bleach for various amounts of time, rinsed with sterile water, and placed on a bacterial culture medium. Clumps of embryogenic tissue which showed no visible bacterial growth after 3 d of culture were then transferred to an agar-solidified plant tissue culture medium to determine viability of bleachdisinfested tissues. Viable, single pieces of the disinfested embryogenic tissue were then used to reinitiate embryogenic suspension cultures. Treatment of contaminated tissue with a 1% bleach solution for 1–5 min resulted in the highest recovery of viable, disinfested tissues using 5 of the 6 bacteria. It was not possible to remove F. balustrum from clumps of embryogenic tissue without also killing the plant tissue.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid Salaries and research support were provided by an Ohio Academy of Sciences/National Science Foundation summer internship award to JAS and by State and Federal funds appropriated to OSU-OARDC. OARDC Journal Article No. 391-89     

Mechanisms of mastoparan-stimulated surfactant secretion from isolated pulmonary alveolar type 2 cells.

Mastoparan, a tetradecapeptide component of wasp venom, is a potent activator of secretion in a variety of cell types, and has been shown to activate purified G-proteins reconstituted into phospholipid vesicles with a preferential activation of Gi over Gs (Higashijima, T., Uzu, S., Nakajima, T., and Ross, E. R. (1988) J. Biol. Chem. 263, 6491-6494). To identify the biochemical activities of mastoparan in a cellular system, we characterized the effects of mastoparan on signal transduction pathways in rat pulmonary alveolar type 2 epithelial cells, which synthesize and secrete pulmonary surfactant. Mastoparan inhibited adenylylcyclase activity in a manner that was dose-dependent (IC50 = 30 microM), but sensitive to neither guanine nucleotide nor pertussis toxin (PT). Mastoparan induced a PT-sensitive increase in cellular inositol trisphosphate and a rapid rise in cytosolic calcium released from intracellular stores; the time to onset of the calcium rise, but neither the rate nor the amplitude of the rise, were PT-sensitive. Mastoparan also caused a dose- (EC50 = 16 microM) and time-dependent activation of arachidonic acid release that was completely insensitive to pretreatment with PT. Secretion of pulmonary surfactant was increased by mastoparan approximately 8-fold over constitutive levels at 1 h with an EC50 = 20 microM, and mastoparan-stimulated secretion was partially sensitive to PT at late time points and to inhibitors of arachidonic acid metabolism, but not to the protein kinase C inhibitor H7. These findings are consistent with the activation of Gi proteins in type 2 cells by mastoparan, although the lack of predicted triphosphoguanine nucleotide and PT sensitivity for some activities indicates that mastoparan does not act in a manner strictly analogous to liganded receptors or that some activities are not mediated by activation of Gi. While mastoparan is a potent secretagogue in several cell types, its secretory activity appears to have only a limited dependence on the activation of Gi proteins in type 2 cells.     

High-resolution profiles of temperature and dissolved oxygen in a river

Longitudinal profiles of water quality along a well-mixed tidal river are, ideally, based on simultaneous measurements at fixed stations distributed along the river. The resolution of the profiles is limited by the density of the stations. However, for a given number of stations the resolution is greatly increased if water quality date can be extrapolated upstream and downstream of the stations, making use of velocity data; the resolution is then determined by the density of the extrapolated data points, which may be an order of magnitude higher than the density of the stations. A 15-km length of river was investigated using 5 current meters equipped to measure depth, temperature, conductivity and dissolved oxygen. Data were recorded simultaneously every 10 minutes. When the average cross-sectional speed was 0.25 ms⁻¹ (typical of tidal conditions), the extrapolated data points were 150 m apart, so the resolution of the resulting profiles (7 per kilometre) was much higher than that of the stations alone (0.3 per kilometre). The extrapolation process required a means of deducing the average cross-sectional speed from the speed measured at a given station. The key to this was provided by temperature data recorded during the onset of a spate, when tidal flow was suspended and the average cross-sectional speed was uniformly about 0.75 ms ⁻¹ at four of the stations. Profiles of temperature and dissolved oxygen were generated by this method; the resolution was about 2 data points km⁻¹ during the onset of the spate, and 6 points km⁻¹ during tidal flow.     

Amino acid uptake and protein synthesis in germinating spores of Saccharomyces cerevisiae.

Spores transferred to germination medium incorporated exogenous lysine into protein within 20 min but required 2-3 to begin incorporation of exogenous proline or alanine. During this time considerable uptake of amino acids into the intracellular pool occurred. Cycloheximide added to the germination medium inhibited incorporation of lysine into protein but did not lessen in accumulation in the pool. Spore germination was inhibited by cycloheximide.     

Characterization of the 3' Termini of the RNAs of Cowpea Mosaic Virus

A sequence of polyadenylic acid, homogeneous in composition but heterogeneous in length, was isolated from complete pancreatic RNase digests of both middle and bottom RNAs of cowpea mosaic virus. The polyadenylic acid was 3′-terminal and occurred once per molecule. A fragment consisting of the polyadenylic acid and approximately the next 25 nucleotides could be isolated from complete T1 RNase digests of either RNA. The region adjacent to the polyadenylic acid in both RNAs was rich in pyrimidines. The mobilities of the fragments in 12.4% polyacrylamide-8 M urea gels were used to estimate their lengths and to calculate number average and weight average molecular weights.