OCCURRENCE OF AN ISOPRENOID C25 DIUNASATURATED ALKENE AND HIGH NEUTRAL LIPID CONTENT IN ANTRACTIC SEA-ICE DIATOM COMMUNITIES1

The lipid and hydrocarbon composition of natural populations of diatom communities collected during the austral spring bloom of 1985 in the sea-ice at McMurdo Sound, Antartica was analyzed by TLC-FID, GC and GC-MS. Sea-ice diatom communities were dominated by Amphiprora sp., Nitzschia stellata Manguin and Berkeleya sp. at Cape Armitage; N. stellata, Amphiprora, Pleurosigma, N. kerguelensis (O'Meara) Hasle and some small centric diatoms adjacent to the Erebus Ice Tongue; and Porosira pseudodenticulata (Hustedt) Jouse at Wohlschlag Bay. Lipid distributions of the sea ice diatom communities from the Cape Armitage and Ereus sites were characterized by high concentrations of tracylaglecycerol (triacylglycerolplar lipid = 1.0 to 1.5). The hydrocarbon n-C_21:6, common in temperate diatoms, and an isoprenoid C₂₅ diunsaturated alkene were the dominant hydrocarons detected at these two sites. Hydrogenation of the C₂₅ diene produced the known alkane 2, 6, 10, 14-tetramethyl-7- (3-methylpentyl)-pentadecane. The C₂₅ diene is one of several structurally related hydrocarbons reported in many estuarine, coastal and ocean ic sediments. We propose that certain species of diatoms are a likely source of these alkenes in sediments. The first reported biological occurrence of the C₂₅ diene in the green seaweed Enteromorpha prolifera may have been due to the presence of epiphytic microalgae in the field sample analysed.     

Microbial biomass in a shallow, urban aquifer contaminated with aromatic hydrocarbons: analysis by phospholipid fatty acid content and composition

The city of Perth contains a number of sites that have been contaminated with hydrocarbons due to leakage from petroleum underground storage tanks. Microbial biomass in groundwater and sediment cores from above and below the water table, and from within and outside a plume of hydrocarbon contamination, was examined using phospholipid fatty acid methyl ester analysis. Microbial numbers, calculated from the phospholipid content, ranged from 0·9 × 10⁶ to 7·8 × 10⁶' Escherichia coli equivalent cells' g⁻¹ dry wt of sediment. Over 96% of the microbial biomass was attached to the sediment and the proportion of attached cells did not decrease within the plume of contaminants. The amount of biomass within aquifer samples seemed to be related more to the proximity of the rhizosphere to the shallow aquifer, and other unknown urban inputs, rather than to the effects of the plume of contaminants. Fatty acids common to many bacterial groups dominated within the plume, and as such the analyses gave limited insight into microbial community structure. For site assessment of intrinsic remediation of shallow aquifers in urban areas, estimates of microbial biomass may not provide information that is readily applicable to plume management.     

Selection of cold‐tolerant plants for growth in soils contaminated with organics

A mixture of organic chemicals (MOC) containing equal molar amounts of benzoic acid, hexadecane, 2,2‐dimethyl 4,n‐propyl‐benzene, phenanthrene, pyrene, and either cycloheptane or cis‐decahydronaphthalene (cis‐decalin) was applied to soil at rates of 0 to 8000 mg/kg. In a plant‐screening experiment, growth responses of four legume and five nonlegume species were determined at 10 and 25°C. The MOC applied at 2000 mg/kg reduced the growth of several species without resulting in significant seedling death. At 10°C, the growth of alpine bluegrass (Poa alpina L.) in the 1000 and 2000 mg/kg treatments of soil increased by more than 185%. In a plant growth response experiment, alpine bluegrass and alfalfa (Medicago sativa L.) were grown in soil that had been contaminated at rates of 0 and 2000 mg/kg. At 14 weeks, the shoot and root dry weights of alfalfa were 97% lower in the contaminated soil, while the shoot dry weight, root dry weight, and root length of alpine bluegrass were 135,235 and 268% higher, respectively. Except for pyrene, <23% of the compounds comprising the MOC remained in the soil after 4 weeks and <5% after 14 weeks. The disappearance of the MOC was not significantly influenced by the presence of alfalfa or alpine bluegrass.     

Melanotic ependymoma in a Goeldi's marmoset (Callimico goeldii)

A spontaneous melanotic ependymoma was observed in the brain of an adult female Goeldi's marmoset (Callimico goeldii). The mass completely occupied the left lateral ventricle, rupturing the fornix and corpus callosum, and compressing the adjacent neuropil. Special histochemical techniques, including melanin bleach, periodic acid-Schiff, Perls iron and phosphotungstic acid hematoxylin, demonstrated the neoplasms to be an ependymoma with a rare melanotic differentiation.     

PcaK, a high-affinity permease for the aromatic compounds 4-hydroxybenzoate and protocatechuate from Pseudomonas putida.

PcaK is a transporter and chemoreceptor protein from Pseudomonas putida that is encoded as part of the beta-ketoadipate pathway regulon for aromatic acid degradation. When expressed in Escherichia coli, PcaK was localized to the membrane and catalyzed the accumulation of two aromatic substrates, 4-hydroxybenzoate and protocatechuate, against a concentration gradient. Benzoate inhibited 4-hydroxybenzoate uptake but was not a substrate for PcaK-catalyzed transport. A P. putida pcaK mutant was defective in its ability to accumulate micromolar amounts of 4-hydroxybenzoate and protocatechuate. The mutant was also impaired in growth on millimolar concentrations of these aromatic acids. In contrast, the pcaK mutant grew at wild-type rates on benzoate. The Vmax for uptake of 4-hydroxybenzoate was at least 25 nmol/min/mg of protein, and the Km was 6 microM. PcaK-mediated transport is energized by the proton motive force. These results show that although aromatic acids in the undissociated (uncharged) form can diffuse across bacterial membranes, high-specificity active transport systems probably also contribute to the ability of bacteria to grow on the micromolar concentrations of these compounds that are typically present in soil. A variety of aromatic molecules, including naturally occurring lignin derivatives and xenobiotics, are metabolized by bacteria and may be substrates for transport proteins. The characterization of PcaK provides a foundation for understanding active transport as a critical step in the metabolism of aromatic carbon sources.     

benK encodes a hydrophobic permease-like protein involved in benzoate degradation by Acinetobacter sp. strain ADP1.

The chromosomal benK gene was identified within a supraoperonic gene cluster involved in benzoate degradation by Acinetobacter sp. strain ADP1, and benK was expressed in response to a benzoate metabolite, cis,cis-muconate. The disruption of benK reduced benzoate uptake and impaired the use of benzoate or benzaldehyde as the carbon source. BenK was homologous to several aromatic compound transporters.     

Regulation of KATP Channel Activity by Diazoxide and MgADP : Distinct Functions of the Two Nucleotide Binding Folds of the Sulfonylurea Receptor

K_ATP channels were reconstituted in COSm6 cells by coexpression of the sulfonylurea receptor SUR1 and the inward rectifier potassium channel Kir6.2. The role of the two nucleotide binding folds of SUR1 in regulation of K_ATP channel activity by nucleotides and diazoxide was investigated. Mutations in the linker region and the Walker B motif (Walker, J.E., M.J. Saraste, M.J. Runswick, and N.J. Gay. 1982. EMBO [Eur. Mol. Biol. Organ.] J. 1:945–951) of the second nucleotide binding fold, including G1479D, G1479R, G1485D, G1485R, Q1486H, and D1506A, all abolished stimulation by MgADP and diazoxide, with the exception of G1479R, which showed a small stimulatory response to diazoxide. Analogous mutations in the first nucleotide binding fold, including G827D, G827R, and Q834H, were still stimulated by diazoxide and MgADP, but with altered kinetics compared with the wild-type channel. None of the mutations altered the sensitivity of the channel to inhibition by ATP⁴⁻. We propose a model in which SUR1 sensitizes the K_ATP channel to ATP inhibition, and nucleotide hydrolysis at the nucleotide binding folds blocks this effect. MgADP and diazoxide are proposed to stabilize this desensitized state of the channel, and mutations at the nucleotide binding folds alter the response of channels to MgADP and diazoxide by altering nucleotide hydrolysis rates or the coupling of hydrolysis to channel activation.     

Movements and associations of ribosomal subunits in a secretory cell during growth inhibition by starvation

In Chironomus tentans salivary gland cells, the cytoplasm can be dissected into concentric zones situated at increasing distances from the nuclear envelope. After RNA labeling, the newly made ribosomal subunits are found in the cytoplasm mainly in the neighborhood of the nucleus with a gradient of increasing abundance towards the periphery of the cell. The gradient for the small subunit lasts for a few hours and disappears entirely after treatment with puromycin. The large subunit also forms a gradient but one which is only partially abolished by puromycin. The residual gradient which which is resistant to the addition of the drug is probably due to the binding of some large ribosomal units to the membranes of the endoplasmic reticulum (J.-E. Edstrom and u. Lonn. 1976. J. Cell Biol. 70:562-572, and U. Lonn and J.-E. Edstrom. 1976. J. Cell. Biol. 70:573-580). If growth is inhibited by starvation, only the puromycin-sensitive type gradient is observed for the large subunit, suggesting that the attachment of these newly made subunits to the endoplasmic reticulum membranes will not occur. If, on the other hand, the drug-resistant gradient is allowed to form in feeding animals, it is conserved during a subsequent starvation for longer periods than in control feeding animals. This observation provides a further support for an effect of starvation on the normal turnover of the large subunits associated with the endoplasmic reticulum. These results also indicate a considerable structural stability in the cytoplasm of these cells worth little or no gross redistribution of cytoplasmic structures over a period of at least 6 days.     

Carbohydrate metabolism and respiration after harvest of the cultivated mushroom, Agaricus bisporus

The carbohydrate metabolism of the mushroom and the respiration of the sporophore after harvest at different stages of growth have been studied. Mannitol and trehalose were found to be the major soluble carbohydrates in the sporophore and mycelium respectively. Mannitol and trehalose levels fall during sporophore storage, and feeding experiments with¹⁴C-labelled sugars indicate that they are metabolised. The results were discussed in relation to the post-harvest development of the sporophore.     

Sequence analysis of the nicks and termini of bacteriophage T5 DNA.

Bacteriophage T5 DNA, when isolated from mature phage particles, contains several nicks in one of the two strands. The 5'-terminal nucleotides at the nicks were labeled with polynucleotide kinase and [gamma-32P]ATP, and the 3'-terminal nucleotides were labeled with Escherichia coli DNA polymerase I and [alpha-32P]dGTP. The sequences around the nicks were analyzed by partial nuclease digestion followed by homochromatography fractionation of the resulting oligonucleotides. The nicks had at least the sequence -PuOH pGpCpGpC- in common. In addition, the two 5' external termini had the first seven nucleotides in common.