DNA Biosynthesis in Chloroplasts: II. DEVELOPMENTAL VARIATION IN TTP INCORPORATION INTO DNA BY PLASTID EXTRACTS

Incorporation of [³H]TTP into DNA by pea chloroplast extractswas highly dependent on the age of the tissue from which plastidswere prepared. Catalytic activity was greatest in samples from6- to 9-d-old plants; preparations from more mature tissueswere much less effective. Moreover, activity was 3 to 10 timesgreater in younger tissues regardless of whether chlorophyll,protein or plastid number was used as the index of comparison.Enzymes from the first (oldest), second, third, and fourth (youngest)leaves of the same plants were also studied. Again, activitywas 4 to 10 times greater in samples from the youngest tissues.When plastid extracts from older leaves were mixed with thosefrom younger tissues, they did not reduce synthesis. Thus, thedecline in activity does not appear to be due to the productionof an inhibitor during plant development. One explanation forthese data is that enzymes of ctDNA replication, such as DNApolymerase, vary in activity during leaf development; thereforechanges in enzyme levels may be an important factor in controllingchloroplast DNA replication during development. We have alsoexamined the incorporation of [³H]TTP into DNA by isolated intactpea chloroplasts; in general, labelled TTP was less readilyincorporated into chloroplast DNA than was [³H]thymidine. Key words: Chloroplast DNA replication, chloroplast DNA polymerase     

Studies of esterase 6 in Drosophila melanogaster. XVIII. Biochemical differences between the slow and fast allozymes

Most natural populations of Drosophila melanogaster are polymorphic for two major electrophoretic variants at the esterase-6 locus. The frequency of the EST 6F allozyme is greatest in populations in warmer latitudes, whereas the EST 6S allozyme is predominant in colder latitudes. Latitudinal clines in electromorph frequencies are found on three continents. Purified preparations of the allozymes have been characterized for their pH optimum, substrate specificity, organophosphate inhibition, alcohol activation, thermal stability, and kinetic parameters. These and previous analyses of the EST 6 allozymes reveal that the two variants have differences in their physical and kinetic properties that may provide a basis for the selective maintenance of the polymorphisms and an explanation of the clinal variation observed in natural populations.      

Sweat gland density and response during high-intensity exercise in athletes with spinal cord injuries

Sweat production is crucial for thermoregulation. However, sweating can be problematic for individuals with spinal cord injuries (SCI), as they display a blunting of sudomotor and vasomotor responses below the level of the injury. Sweat gland density and eccrine gland metabolism in SCI are not well understood. Consequently, this study examined sweat lactate (S-LA) (reflective of sweat gland metabolism), active sweat gland density (SGD), and sweat output per gland (S/G) in 7 SCI athletes and 8 able-bodied (AB) controls matched for arm ergometry VO₂peak. A sweat collection device was positioned on the upper scapular and medial calf of each subject just prior to the beginning of the trial, with iodine sweat gland density patches positioned on the upper scapular and medial calf. Participants were tested on a ramp protocol (7 min per stage, 20 W increase per stage) in a common exercise environment (21±1°C, 45-65% relative humidity). An independent t-test revealed lower (p<0.05) SGD (upper scapular) for SCI (22.3 ±14.8 glands · cm⁻²) vs. AB. (41.0 ± 8.1 glands · cm⁻²). However, there was no significant difference for S/G between groups. S-LA was significantly greater (p<0.05) during the second exercise stage for SCI (11.5±10.9 mmol · l⁻¹) vs. AB (26.8±11.07 mmol · l⁻¹). These findings suggest that SCI athletes had less active sweat glands compared to the AB group, but the sweat response was similar (SLA, S/G) between AB and SCI athletes. The results suggest similar interglandular metabolic activity irrespective of overall sweat rate.     

The molar dentition of a Welsh pantothere

The paper describes the molar teeth of a primitive therian mammal from the early Jurassic of S. Wales. The teeth are all isolated with largely intact crowns but with the roots usually broken. An attempt is made to reproduce a molar series and it is suggested that the animal has five constant molars with a sixth which was inconstant in size and probably not always present. The teeth exhibit a number of very primitive features. The animal may well be congeneric with Kuehneotherium praecursoris , from which it differs in a number of details.     

Chronic exposure to increasing background ozone impairs stomatal functioning in grassland species

Two species found in temperate calcareous and mesotrophic grasslands (Dactylis glomerata and Leontodon hispidus) were exposed to eight ozone treatments spanning preindustrial to post‐2100 regimes, and late‐season effects on stomatal functioning were investigated. The plants were grown as a mixed community in 14 L containers and were exposed to ozone in ventilated solardomes (dome‐shaped greenhouses) for 20 weeks from early May to late September 2007. Ozone exposures were based on O₃ concentrations from a nearby upland area, and provided the following seasonal 24 h means: 21.4, 39.9 (simulated ambient), 50.2, 59.4, 74.9, 83.3, 101.3 and 102.5 ppb. In both species, stomatal conductance of undamaged inner canopy leaves developing since a midseason cutback increased linearly with increasing background ozone concentration. Imposition of severe water stress by leaf excision indicated that increasing background ozone concentration decreased the ability of leaves to limit water loss, implying impaired stomatal control. The threshold ozone concentrations for these effects were 15–40 ppb above current ambient in upland UK, and were within the range of ozone concentrations anticipated for much of Europe by the latter part of this century. The potential mechanism behind the impaired stomatal functioning was investigated using a transpiration assay. Unlike for lower ozone treatments, apparently healthy green leaves of L. hispidus that had developed in the 101.3 ppb treatment did not close their stomata in response to 1.5 μm abscisic acid (ABA); indeed stomatal opening initially occurred in this treatment. Thus, ozone appears to be disrupting the ABA‐induced signal transduction pathway for stomatal control thereby reducing the ability of plants to respond to drought. These results have potentially wide‐reaching implications for the functioning of communities under global warming where periods of soil drying and episodes of high vapour pressure deficit are likely to be more severe.     

Characterization of the genetic diversity of <Emphasis Type="Italic">Mycobacterium tuberculosis</Emphasis> in São Paulo city,Brazil

Background

Tuberculosis is a major health problem in São Paulo, Brazil, which is the most populous and one of the most cosmopolitan cities in South America. To characterize the genetic diversity of Mycobacterium tuberculosis in the population of this city, the genotyping techniques of spoligotyping and MIRU were applied to 93 isolates collected in two consecutive years from 93 different tuberculosis patients residing in São Paulo city and attending the Clemente Ferreira Institute (the reference clinic for the treatment of tuberculosis).

Findings

Spoligotyping generated 53 different spoligotype patterns. Fifty-one isolates (54.8%) were grouped into 13 spoligotyping clusters. Seventy- two strains (77.4%) showed spoligotypes described in the international databases (SpolDB4, SITVIT), and 21 (22.6%) showed unidentified patterns. The most frequent spoligotype families were Latin American Mediterranean (LAM) (26 isolates), followed by the T family (24 isolates) and Haarlem (H) (11 isolates), which together accounted for 65.4% of all the isolates. These three families represent the major genotypes found in Africa, Central America, South America and Europe. Six Spoligo-International-types (designated SITs by the database) comprised 51.8% (37/72) of all the identified spoligotypes (SIT53, SIT50, SIT42, SIT60, SIT17 and SIT1). Other SITs found in this study indicated the great genetic diversity of M. tuberculosis, reflecting the remarkable ethnic diversity of São Paulo city inhabitants. The MIRU technique was more discriminatory and did not identify any genetic clusters with 100% similarity among the 93 isolates. The allelic analysis showed that MIRU loci 26, 40, 23 and 10 were the most discriminatory. When MIRU and spoligotyping techniques were combined, all isolates grouped in the 13 spoligotyping clusters were separated.

Conclusions

Our data indicated the genomic stability of over 50% of spoligotypes identified in São Paulo and the great genetic diversity of M. tuberculosis isolates in the remaining SITs, reflecting the large ethnic mix of the São Paulo city inhabitants. The results also indicated that in this city, M. tuberculosis isolates acquired drug resistance independently of genotype and that resistance was more dependent on the selective pressure of treatment failure and the environmental circumstances of patients.

     

Changes in the timing of spring and autumn migration in North American migrant passerines during a period of global warming

Butler (2003) used first arrival dates (FADs) of 103 migrant birds in northeastern USA and found that both long-distance migrants (LDMs; wintering south of the USA) and short-distance migrants (SDMs; wintering in the southern USA) arrived earlier in the second half of the 20th century than they had in the first, consistent with scenarios of global warming; the trend was stronger in SDMs. Using FADs to characterize migration systems can be problematic because they are data from one tail of a distribution, they comprise a mostly male population and they may not correlate well with the balance of the migration period. FADs also provide no information about autumn migration. This paper uses a banding dataset from Long Point Bird Observatory, Ontario, for 14 passerines for a period of global warming (1975–2000), taking these issues into account. The data were filtered to minimize effects of unequal netting effort (147 491 resulting records), and the passage dates then calculated in each season of each year for the 1st, 2nd and 3rd quartiles for regression analysis. Only two of 13 species analysed in the spring showed significantly earlier passage times, although the overall trend was towards earlier spring migration, especially among SDMs. Autumn responses were more prevalent, however, and in some cases more dramatic with six of 13 species showing delayed migration (four SDMs, two LDMs). Two LDMs exhibited earlier autumn migration. Where earlier spring migration occurred, both sexes appeared to contribute to the change. Where delayed migration occurred in autumn, both sexes and both adults and hatch-year birds appeared to contribute in at least some cases. The spring FAD results are consistent with those of Butler, but when the whole migration is considered, change is far from universal in spring and is in fact more substantial and complex in autumn.