Partial duplication of the large ribosomal RNA sequence in an inverted repeat in circular mitochondrial DNA from Kloeckera africana: Implications for mechanisms of the petite mutation

The 27,100 base-pair circular mitochondrial DNA from the yeast Kloeckera africana has been found to contain an inverted duplication spanning 8600 base-pairs. Sequences hybridizing to transfer RNAs and the large ribosomal RNA are present in the duplication; however, one end of this segment terminates in the large mitochondrial ribosomal RNA sequence so that at least 1000 base-pairs of the gene are not repeated. The large and small mitochondrial ribosomal RNAs have been shown to have lengths of 2700 and 1450 bases, respectively, and genes for these sequences are separated by a minimum of 1300 base-pairs and a maximum of 1750 base-pairs. Consequences of the large inverted duplication to mechanisms of the petite mutation are discussed in terms of previous hypotheses centred on intramolecular recombination in yeast mitochondrial DNA at sequences of homology or partial homology. Despite the long inverted duplication in K. africana mitochondrial DNA, this yeast has one of the lowest frequencies of spontaneous petite mutants amongst petite positive yeasts. One implication of these findings is that in this yeast intra-molecular mitochondrial DNA sequence homology may not be an important factor in the excision process leading to petite formation.     

In vivo phytochrome reversion in immature tissue of the alaska pea seedling

Reversion of far red-absorbing phytochrome to red-absorbing phytochrome without phytochrome destruction (that is, without loss of absorbancy and photoreversibility) occurs in the following tissues of etiolated Alaska pea seedlings (Pisum sativum L.): young radicles (24 hours after start of imbibition), young epicotyls (48 hours after start of imbibition), and the juvenile region of the epicotyl immediately subjacent to the plumule in older epicotyls. Reversion occurs rapidly in the dark during the first 30 minutes following initial phototransformation of red-absorbing phytochrome to far red-absorbing phytochrome. If these tissues are illuminated continuously with red light for 30 minutes, the total amount of phytochrome remains unchanged. Beyond 30 minutes after a single phototransformation or after the start of continuous red irradiation, phytochrome destruction commences. In young radicles, sodium azide inhibits this destruction, but does not affect reversion. In older tissues in which far red-absorbing phytochrome destruction begins immediately upon phototransformation, strong evidence for simultaneous far red-absorbing phytochrome reversion is obtained from comparison of far red-absorbing phytochrome loss in the dark following a single phototransformation with far red-absorbing phytochrome loss under continuous red light.     

Notes on the use of wood refugia by Physella heterostropha (say) in a thermally disturbed swamp

Physella heterostropha size distributions and densities were measured in Pen Branch delta, a thermally altered swamp of the Savannah River, South Carolina. During cessation of thermal input, measurements were taken in a highly impacted area and more natural area. Of the three substrates sampled, logs, small woody material and benthos, snail density was highest on small pieces of submerged wood and < 20% of the snails on logs were out of the water. Following resumption of thermal input, snail density was four times higher on logs than previously measured. Snails responded to elevated water temperatures by congregating in a narrow band slightly above the water. Snail size did not appear to affect their response. However, the thermal history of the snails influenced their behaviour and survival rates. Logs may be a potential refuge for snails when water temperature are greater greater than 38°C. However, our results indicate that long-term survival in this manner may be impossible.     

The allometry of algal respiration

For 30 years, study after study has shown that respiration ratesincrease as {small tilde}0.75 of body size for organisms rangingfrom protozoans to mammals. However, a number of studies suggestedthat the respiration-size relationship for algae may be a rareexception to this general rule. Algal respiration may be almostproportional to cell size, such that the slope of the respiration-sizerelationship is closer to unity. The present study examinedthe effect of cell size and taxon on phytoplankton respiration,using data collected from the literature. To this end, we collecteda data set of 178 observations of algal respiration and cellsize representing six divisions-chlorophytes. chrysophytes,cyanophytes, euglenophytes, pyrrophytes and rhodophytes. Therelationship between respiration (R, in p1 O₂ cell^–1 h^–1)and cell carbon content (C, in pg C cell^–1) is describedas R = 0.030C^{0 93} and the exponent is significantly >3/4.When we expressed cell size in terms of volume, the exponentdecreased to 0.88 but this is still significantly >3/4. Amongthe six divisions studied, chlorophytes, euglenophytes and rhodophytesseemed to differ significantly in their respiration-size relationshipfrom other taxa. However, euglenophytes and rhodophytes havesuch small size ranges that no meaningful relationships canbe developed for those groups alone. The chlorophyte respiration-sizerelationship has obvious patterns in its residuals which mayindicate that significant sources of error were not controlledin these heterogeneous data. Thus, for the present, the generalmodel seems most appropriate for the prediction of respirationrates of phytoplankton.     

Light effects upon dry lettuce seeds

Germination of certain dry seeds (achenes) of Lactuca sativa L. cv. Grand Rapids was increased to ca. 75% after irradiation with 665 nm red light (R; 1x10³ J m^-2); this response was eliminated by far-red light (FR) following the R. The response of dry seeds required an order of magnitude more light than that of wetted seeds, and was not maximal until 48 h after irradiation. Other seeds, which could not be stimulated by R in dry state, showed a partial response after 10 min hydration. Irradiation of dry seeds (or seeds wetted 1 h) with FR (1x10³ J m^-2) reduced dark germination from 26% to 2%. Seeds dehydrated in an oven (60°C, 90 min) showed a decrease in germination if irradiated with R (1x10⁵ J m^-2) before wetting. The results show that phytochrome is present in dry lettuce seeds (and functional in some seed lots) prior to wetting; and that in other seed lots the molecule becomes functional within minutes after wetting the seeds. Transformation of the FR absorbing from of phytochrome (P_FR) to the inactive from (P_R) occurs at lower seed moisture content than the reverse reaction. It appears that dormancy in seeds ripened in sunlight might be assured during seed drying and maturation by the more effective transformation of P_FR to P_R than vice versa as phytochrome is dehydrated.Abbreviations FR far-red - R red - CAL seeds from California - NC seeds from North Carolina (see text)     

Management of diabetes mellitus in children.

In many respects the management of diabetes mellitus in children differs from the management in adults. Guidelines for the short- and long-term management of the child with diabetes mellitus, particularly as it relates to clinical presentation, treatment of ketoacidosis, long-term therapy and psychological counseling, are presented. The specific aspects of diabetes management that are unique to the child are best met by an increased understanding of the problems by the child and the family and an integrated approach by the physician and allied health personnel.     

Recovery of novel bacterial diversity from a forested wetland impacted by reject coal

Sulphide mineral mining together with improperly contained sulphur-rich coal represents a significant environmental problem caused by leaching of toxic material. The Savannah River Site's D-area harbours a 22-year-old exposed reject coal pile (RCP) from which acidic, metal rich, saline runoff has impacted an adjacent forested wetland. In order to assess the bacterial community composition of this region, composite sediment samples were collected at three points along a contamination gradient (high, middle and no contamination) and processed for generation of bacterial and archaeal 16S rDNA clone libraries. Little sequence overlap occurred between the contaminated (RCP samples) and unimpacted sites, indicating that the majority of 16S rDNAs retrieved from the former represent organisms selected by the acidic runoff. Archaeal diversity within the RCP samples consisted mainly of sequences related to the genus Thermoplasma and to sequences of a novel type. Bacterial RCP libraries contained 16S rRNA genes related to isolates (Acidiphilium sp., Acidobacterium capsulatum, Ferromicrobium acidophilium and Leptospirillum ferrooxidans) and environmental clones previously retrieved from acidic habitats, including ones phylogenetically associated with organisms capable of sulphur and iron metabolism. These libraries also exhibited particularly novel 16S rDNA types not retrieved from other acid mine drainage habitats, indicating that significant diversity remains to be detected in acid mine drainage-type systems.     

Next-Generation Site-Directed Transgenesis in the Malaria Vector Mosquito Anopheles gambiae: Self-Docking Strains Expressing Germline-Specific phiC31 Integrase

Diseases transmitted by mosquitoes have a devastating impact on global health and the situation is complicated due to difficulties with both existing control measures and the impact of climate change. Genetically modified mosquitoes that are refractory to disease transmission are seen as having great potential in the delivery of novel control strategies. The Streptomyces phage phiC31 integrase system has been successfully adapted for site-directed transgene integration in a range of insects, thus overcoming many limitations due to size constraints and random integration associated with transposon-mediated transformation. Using this technology, we previously published the first site-directed transformation of Anopheles gambiae, the principal vector of human malaria. Mosquitoes were initially engineered to incorporate the phiC31 docking site at a defined genomic location. A second phase of genetic modification then achieved site-directed integration of an anti-malarial effector gene. In the current publication we report improved efficiency and utility of the phiC31 integrase system following the generation of Anopheles gambiae self-docking strains. Four independent strains, with docking sites at known locations on three different chromosome arms, were engineered to express integrase under control of the regulatory regions of the nanos gene from Anopheles gambiae. The resulting protein accumulates in the posterior oocyte to provide integrase activity at the site of germline development. Two self-docking strains, exhibiting significantly different levels of integrase expression, were assessed for site-directed transgene integration and found to demonstrate greatly improved survival and efficiency of transformation. In the fight against malaria, it is imperative to establish a broad repertoire of both anti-malarial effector genes and tissue-specific promoters to regulate their expression, enabling those offering maximum effect with minimum fitness cost to be identified. The improved technology we describe here will facilitate comparative studies of effector transgenes, allowing informed choices to be made that potentially lead to transmission blockade.