The Effect of Oxygen on the Frequency of Somatic Recombination in DROSOPHILA MELANOGASTER

The influence of oxygen on the frequency of somatic recombination in the yellow singed system on the X chromosome of Drosophila melanogaster was studied under a variety of experimental conditions. Flies raised from egg to adult in atmospheres containing 70-90% oxygen were found to have significantly more mosaic spots on their abdominal tergites than were observed in flies which developed in air. First instar larvae X-rayed in from 0 to 100% oxygen demonstrated the existence of an oxygen effect for somatic recombination in the cells which form the abdominal hypoderm. The mosaic spot counts, beginning with the lowest numbers which were found in flies X-rayed in nitrogen, increased rapidly with rising oxygen tensions until the percentage in air was reached, then leveled off at the higher concentrations. Post-treatment with nitrogen of larvae X-rayed in air or oxygen created a substantially higher number of mosaic spots than were found when larvae, after being similarly irradiated, were instead placed into air or oxygen.     

Excision of one of two defective P elements as the cause of alternate mutational events (sn+ and sne) of the singed bristle allele snw in Drosophila melanogaster

The X-linked singed locus is concerned with the bristle phenotype and female sterility, and is known as a hot spot of P element insertion. A moderate allele of singed, singed-weak (snw) (Engels, 1979; 1984) is inserted with P elements. It is used as an index of P element activity, since it mutates at a high frequency to either a more extreme allele, singed-extreme (sne), or to a phenotype that is equivalent to the wild type (sn+) when an autonomous P element exists. We show here that snw is inserted with two defective P elements in reverse orientation, and the two alternate mutational events (sn+ and sne) are caused by the excision of one or the other of the P elements present in the singed gene. It is interesting that sn+ and sne are inserted with a single P element in the same position, but show very different phenotypes. The insertional sites of P elements in the singed locus possibly contain an unidentified repetitive sequence, which is repeated dozens of times per haploid genome of the wild-type strain Canton-S.     

Changes in the composition of intracellular lipids and hydrocarbons in Candida tropicalis in the absence of growth]

The fatty acid composition of lipids and the composition of hydrocarbons were studied in Candida tropicalis cultivated on a medium with propionic acid and incubated in the conditions of starvation and on a medium containing glucose-1-6-14C but no nitrogen. Intracellular fatty acids with an odd number of carbon atoms were found to be easily metabolized by yeast cells. The content and composition of intracellular hydrocarbons were very labile and underwent intensive transformation caused by changes in the metabolism of the yeast cell.     

Influence of different vitamin-nitrogen sources on cell growth and propionic acid production from sucrose by Propionibacterium shermanii

Cell growth and organic acid production by Propionibacteria are dependent on the vitamin-nitrogen source in the culture medium. Final cell and propionic acid concentrations produced by Propionibacterium shermanii, using corn-steep liquor, were higher than those obtained utilizing yeast extracts. Since corn-steep liquor is much cheaper than yeast extract, the process becomes more attractive. By calculating the specific growth rates, it was observed that the critical propionic acid concentration, that prevents all growth (μ_X = 0), is different depending on the vitamin-nitrogen source used and its concentration. For example, for 5.0 and 15.0 g/l Oxoid yeast extract, those critical propionic acid concentrations were 16.0 and 27.0 g/l, respectively. Such propionic acid concentrations inhibit the cell growth, but not the formation of acid. The specific propionic acid production rate also indicates that the critical concentration for metabolic activity, when propionic acid is no longer produced (μ_P = 0), varies according to the vitamin-nitrogen source and its concentration in the medium. For 5.0 and 15.0 g/l Oxoid yeast extract, those concentrations were 22.1 and 30.1 g/l, respectively.     

废糟液全循环条件下絮凝酵母乙醇连续发酵

丙酸是以玉米为原料自絮凝酵母乙醇连续发酵系统废糟液全循环过程中积累的主要抑制物。基于丙酸对酵母细胞抑制机理,开发了3种废糟液全循环条件下乙醇连续发酵工艺策略。首先根据高温导致丙酸生成的现象,去除了物料灭菌环节,使发酵液丙酸浓度显著降低,生物量和乙醇浓度分别提高了59.3%和7.4%。其次,以丙酸浓度达到半数抑制浓度(IC50)40 mmol/L为目标,通过拟合丙酸积累数据预测废糟液全循环的最长运行时间,发酵装置运行应控制在此时间范围内。再次,较低的环境pH值提高了丙酸毒性,而实验证明发酵液pH为5.5时,丙酸对细胞生长的抑制影响最小,因此控制发酵过程中的pH有利于弱化丙酸毒性。     

Superunstable systems in Drosophila melanogaster. Analysis of mutations in signed and cut loci

The lines of the M'-cytotype characterized by a long-term instability (which was shown to be conditioned by transpositions of the new mobile element, Stalker) were hybridized with the P-line. This resulted in the appearance of a number of superunstable mutations at the yellow, white, singed, ocelliless and some other loci. The authors analyzed four independently obtained families of superunstable mutations at the singed locus. A wide spectrum of derivatives and high frequency of mutations were demonstrated, as well as the regularities of allelic transitions. Besides this, mutagenesis at the cut locus was observed in the chromosomes carrying sn mutations with frequency of 5.05 x 10(4). By means of the blot analysis it has been shown that most of ct mutations are intragenic deficiencies, ranging from 1.3 to 3 Kb, whose appearance is, conceivably, attributed to the inaccuracy of the insertion excision (the insertion is present but fails to alter the phenotype) at the cut locus of the chromosomes with the superunstable sn-alleles. In the lines with the sn- and ct-mutations the transpositions of the P-element and the Stalker were found, which indicates their involvement in mutagenesis. The authors discuss possible effects of inserting the complicated constructions, based on the combinations of P-element and the Stalker, on the induction of superinstability.     

Interaction of mobile elements P and mdg3 in Drosophila melanogaster: genetic aspects

It was found earlier that two unstable sn mutants isolated from natural populations are connected with insertion of mobile element mdg3 into the 7D1-2 region where singed gene (1-21.0) is localised. From two original sn mutants, a series of unstable sn alleles, both mutant and normal for phenotype, was extracted. Then we studied, how they change the mutation rate in germinal and somatic cells of different hybrids with pi 2 stock having P cytotype and active P elements in the chromosomes. Addition of P chromosomes, independently of the background of cytoplasm, proved to reduce the sn instability. The level of sn mutability was decreased with increasing the dose of P chromosomes. It is suggested that mutation events are caused by transposition of mdg3 and that both mdg3 and P elements compete for the same cellular factor, capable of activation of transposition process.     

Development in Genetic Mosaics of Aristapedia, a Homoeotic Mutant of DROSOPHILA MELANOGASTER

Development of the homoeotic mutation, aristapedia (ss(a)), was investigated by means of genetic mosaics. The wild-type alleles of aristapedia and the bristle markers yellow, singed, and forked were removed from cells at different times in development by X-ray induced somatic crossing-over. The phenotype of the resulting clones was examined in order to ascertain whether it was leg or antenna. The y sn f; ss(a) clones showed a leg phenotype if induced before the mid-third instar, but showed an antennal phenotype if induced after this time. Late non-expression of ss(a) may be due either to an influence of surrounding ss(+) tissues on the small ss(a) clones, or to a persistence of the effect of ss(+) for one or two cell generations after it is removed from a cell line.     

A novel pathway for transport and metabolism of a fluorescent phosphatidic acid analog in yeast

Phosphatidic acid is a central intermediate of biosynthetic lipid metabolism as well as an important signaling molecule in the cell. These studies assess the internalization, or retrograde transport , and metabolism of phosphatidic acid in yeast using a fluorescent analog. An analog of phosphatidic acid fluorescently labeled at the sn -2 position with N-4-nitrobenz-2-oxa-1, 3-diazole-aminocaproic acid (NBD-phosphatidic acid) was introduced to yeast cells by spontaneous transfer from phospholipid vesicles. Transport and metabolism of the NBD-phosphatidic acid were then monitored by fluorescence spectrophotometry, fluorescence microscopy and routine biochemical methods. Primary metabolites of the NBD-phosphatidic acid in yeast were found to be NBD-diacylgycerol and NBD-phosphatidylinositol. Experiments in cells possessing different levels of phosphatidate phosphatase activity suggest that conversion of the NBD-phosphatidic acid to NBD-diacylglycerol is not a pre-requisite for internalization in yeast. Internalization is sensitive to decreased temperature, but neither ATP depletion nor a sec6-4 mutation, which interrupts endocytosis, has an affect. Thus, internalization of NBD-phosphatidic acid apparently occurs via a non-endocytic route. These characteristics of retrograde transport of NBD-phosphatidic acid in yeast differ significantly from transport of other NBD-phospholipids in yeast as well as NBD-phosphatidic acid transport in mammalian fibroblasts.     

Mutagenic effects of some water-soluble metal compounds in a somatic eye-color test system in Drosophila melanogaster

Nickel, cadmium, lead, arsenic, manganese and chromium salts as well as MeHgOH were screened for mutagenicity, using a sensitive somatic eye-color test system in Drosophila melanogaster. The test is based on the insertion of a mobile element which causes instability in the white locus that is somatically enhanced by mutagens. This white locus expression is combined with a mutation, zeste, in another gene, to produce a light yellow eye color. Larval feeding with mutagens causes somatic mutations in the eye imaginal disc cells that develop into easily detectable red spots in the yellow eyes of adult males. Survival tests showed large differences in the toxicity of different metals, but only hexavalent chromium increased the frequency of somatic mutations above the control level. When combined treatments were carried out with MMS and various metals, sodium arsenite caused a reduction of the MMS-induced mutation frequency while methylmercury increased the frequency of somatic spots.     

Ophioderma peruana,a new species of brittlestar (Echinodermata,Ophiuroidea, Ophiodermatidae) from?the Peruvian coast

Ophioderma peruana sp. n. is a new species of Ophiodermatidae, extending the distribution of the genus Ophioderma to Lobos de Afuera Island, Peru, easily distinguishable from its congeners by its peculiarly fragmented dorsal arm plates. Dense granules, rounded or polygonal cover the disc, the radial shields may be naked or completely covered by granules. A good character for recognizing this species in the field is the dorsal side of the disc which is brown with disc granules lighter cream and brown, the arms are mottled with whitish spots and the ventral part of the disc on the interradial part is brown and the radial part bright yellow.     

Active mariner transposable elements are widespread in natural populations of Drosophila simulans

The occurrence of active, or autonomous, mariner elements was investigated by crossing white-peach mutant Drosophila simulans females with wild-type males from various geographic origins. From a total of 194 experimental crosses only 17 failed to produce progeny with eye mosaicism (MOS, i.e. pigmented spots in otherwise white-peach eyes). Therefore, active mariner elements inducing somatic excision of the copy inserted at the white locus are abundant in all populations sampled. In the experimental crosses the frequency of mosaic offspring ranged from 0 to 100%, showing that the phenotypic expression is highly variable. The MOS phenotype, measured by the number of spots on the eyes, is quite variable within the progeny of single crosses. Although a difference was observed in the average MOS score (percentage of mosaic flies) between northern and southern populations of France, there was no indication of long range variation between geographic populations. Neither was there a systematic difference between recently collected populations and samples kept several years as isofemale lines.     

Effect of endocytosis upon acid phosphatase activity ofTetrahymena pyriformis

Summary Increased endocytosis inTetrahymena pyriformis, produced by presenting starved cells with either peptone-yeast extract medium or killed yeast cell suspension, results in increased cellular acid phosphatase activity.Tetrahymena, grown in peptone-yeast extract medium, showed increased acid phosphatase activity after phagocytosis of yeast cells. This increase was not apparent until about one hour after presentation and was maximal at about 2.5 hours.Tetrahymena, grown on yeast suspension, showed little increase in acid phosphatase activity on presentation with peptone-yeast extract medium. These results may indicate that endocytosis, of either particles or solutes, produces an adaptive increase in acid phosphatase activity (presumably lysosomal in nature) which is related to feeding.Histochemical examination failed to localise the increase in acid phosphatase activity cellularly, but small particles, of about 1 diameter, which showed acid phosphatase activity and were presumably lysosomes were noted. Closely orientated yeast cells showed varying intensities of lead deposition, from absence to intense staining. This suggests that newly ingested yeast cells may be ingested initially in a single phagosome and that thereafter one or more lysosomes may fuse with them.     

Propionic acid fermentation of ultra-high-temperature sterilized whey using mono-and mixed-cultures

Summary Unlike sterilization by autoclave (Anderson et al. 1986) high concentrations of cheese whey sterilized by ultra high temperature (UHT) resulted in a medium conducive to microbial growth and propionic acid production. Propionibacterium freudenreichii ss. shermanii, grown with pH control in 12% whey solids and 1% yeast extract sterilized by UHT, produced about 1.9% propionic acid within 70 h; more than 50% of the lactose was not used. Under similar conditions, mixed cultures of P. shermanii and Lactobacillus casei produced more than 3.0% propionic acid. Acclimating the mixed culture to the whey medium resulted in 4.5% propionic acid. The amount of propionic acid produced was further increased to about 6.5% by raising the concentration of whey solids to about 18%. Using the mixed culture, all the lactose was consumed and lactic acid did not accumulate.     

Effects of Rapeseed Oil on Activity of Methylmalonyl-CoA Carboxyltransferase in Culture of Streptomyces fradiae

To investigate why more tylosin was produced when Streptomyces fradiae T1558 was cultured in a rapeseed oil medium than in a glucose or starch medium, we measured the activity of methylmalonyl-CoA carboxyltransferase (EC 2.1.3.1) and intracellular propionic acid. The activity of the enzyme, which catalyzes the formation of the precursor of tylosin, protylonolide, was 0.19 U/mg protein in 5 days of culture in rapeseed oil medium, which was 2.5- and 1.3-fold that with the glucose or starch medium, respectively. The intracellular propionic acid concentration was 1.2 g/g of dry weight, which was 4.3- and 2.1-fold that with the glucose or starch medium, respectively. The addition of propionic acid increased tylosin production in batch culture: when 0.2 g/l (final concentration) propionic acid was added to the glucose medium, 3.8 g/l tylosin was produced in 10 days of culture, 4.7-fold the amount without propionic acid. These findings suggest that in glucose medium, intracellular propionic acid is a limiting factor because of the low activity of methylmalonyl-CoA carboxyltransferase of the tylosin biosynthesis pathway.     

Molecular Analysis of an Unstable P Element Insertion at the Singed Locus of Drosophila Melanogaster: Evidence for Intracistronic Transposition of a P Element

In a companion study, a number of P element insertions into the singed locus were characterized. Here is reported a detailed analysis of the structure and mutability of another P element insertion at sn, known as sncm. Under conditions which mobilize P elements, sncm mutates at high frequency to both wild-type (sn+) and to a much more extreme allele (snext). Wild-type revertants appear to represent precise or nearly precise excisions of the P element. Certainly two, and most likely all five, of the snext alleles studied result from the insertion of a duplicate copy of this P element into a nearby site in an inverted orientation. We propose a model in which both the sn+ and snext mutational events can be explained by excision of the P element from one chromatid followed by reintegration into the sister chromatid at a nearby site (intracistronic transposition). Finally, it is shown that the snext alleles are themselves unstable and the structure of a resulting chromosome aberration is examined.     

The defective dorsal discs gene of Drosophila is required for the growth of specific imaginal discs

The wild-type allele of the gene defective dorsal discs (ddd) is required for the normal development of the dorsal thoracic discs in Drosophila melanogaster. In ddd mutant larvae the dorsal discs (wing, haltere, and humeral) are greatly reduced in size or absent while the ventral discs (leg) are unaffected. We have examined the function of the ddd+ gene in wing development. The ddd+ product is not involved in the initial determination of wing cells but rather is required for their subsequent proliferation during the larval period. Analysis of chimaeras shows that there is a requirement for ddd+ gene expression in wing discs, but it is sufficient for normal development that only some cells in a disc express the gene. We propose that the ddd+ product is involved in the synthesis of a factor which is required for the normal growth of wing discs and which can be transferred between wing disc cells.     

Anti-yeast activity of a food-grade dilution-stable microemulsion

The anti-yeast activities of a food-grade dilution-stable microemulsion against Candida albicans and Saccharomyces cerevisiae have been studied. The weight ratio of the formulated microemulsion is glycerol monolaurate (GML)/propionic acid/Tween 80/sodium benzoate (SB)/water = 3:9:14:14:24. Results of anti-yeast activity on solid medium by agar diffusion method showed that the anti-yeast activity of the microemulsion at 4.8 mg/ml was comparable to that of natamycin at 0.1 mg/ml as positive control. Results of anti-yeast activity in liquid medium by broth dilution method showed that the growth of both C. albicans and S. cerevisiae was completely inhibited when the liquid medium containing 10⁶ cfu/ml was treated with 1.2 mg/ml microemulsion, which was determined as minimum fungicidal concentration. The kinetics of killing results showed that the microemulsion killed over 90% yeast cells rapidly within 15 min and caused a complete loss of viability in 120 min. Among the components, SB and GML had a similar anti-yeast activity, followed by propionic acid, while Tween 80 exhibited no activity and could not enhance the anti-yeast activities of these components, and it was revealed that the anti-yeast activity of the microemulsion was attributed to a combination of propionic acid, GML, and SB. The anti-yeast activity of the microemulsion was in good agreement with the leakage of 260-nm absorbing materials and the observation of transmission electron microscopy, indicating that the microemulsion induced the disruption and dysfunction of the cell membrane.