In vitro studies of the submerged angiospermRuppia maritima: Auxin and cytokinin effects on plant growth and development

Axenic tissue cultures ofRuppia maritima L. were established and propagated clonally in vitro from terminal rhizome segments collected from Tampa Bay, Florida, USA. Cultures were maintained in a base medium consisting of synthetic seawater supplemented with half-strength Murashige and Skoog salts and 1% sucrose at pH 5.6. The effects of five cytokins [6-furfurylaminopurine (kinetin), 6-benzylaminopurine (BAP), 2-isopentyladenine (2iP), 6-(4-hydroxy-3-methyl-but-2-enylamino) purine (zeatin), andn-phenyl-n-1,2,3-thidiazol-5yl urea (thidiazuron)] and one auxin [napthalene acetic acid (NAA)] on explant growth and development were investigated. Cytokinin additions resulted in a 3- to 4-fold increase in nodal production, branching, and biomass ofR. maritima after 12 wk in culture. Cultures responded in a dose-dependent manner to 2iP but exhibited broad dose-response curves to kinetin, BAP, zeatin, and thidiazuron. NAA addition resulted in increased leaf and internodal lengths, but reduced the number of leaves per node and the rhizome biomass. The addition of NAA almost completely suppressed root growth in media without cytokinins and had an antagonistic effect on nodal production and branching in cytokinin-supplemented media.     

Interlaboratory evaluation of endotoxin analyses in agricultural dusts--comparison of LAL assay and mass spectrometry

Endotoxin exposure is associated with wheeze and asthma morbidity, while early life exposure may reduce risk of allergy and asthma. Unfortunately, it is difficult to compare endotoxin results from different laboratories and environments. We undertook this study to determine if lipopolysaccharide (LPS) extraction efficiency could account for differences among laboratories. We generated and collected aerosols from chicken and swine barns, and corn processing. We randomly allocated side-by-side filter samples to five laboratories for Limulus assay of endotoxin. Lyophilized aliquots of filter extracts were analyzed for 3-hydroxy fatty acids (3-OHFAs) as a marker of LPS using gas chromatography-mass spectrometry. There were significant differences in endotoxin assay and GC-MS (LPS) results between laboratories for all dust types (p < 0.01). Patterns of differences between labs varied by dust type. Relationships between assay and GC/MS results also depended on dust type. The percentages of individual 3-OHFA chain lengths varied across labs (p < 0.0001) suggesting that each lab recovered a different fraction of the LPS available. The presence of large amounts of particle associated LPS and absence of a freezing thawing cycle were associated with lower correlations between LPS and bioactivity, consistent with an absence of Limulus response to cell-bound endotoxin. These data suggest that extraction methods affect endotoxin measurements. The LAL methods may be most suitable when comparing exposures within similar environments; GC-MS offers additional information helpful in optimizing sample treatment and extraction. GC-MS may be of use when comparing across heterogeneous environments and should be considered for inclusion in future studies of human health outcomes.     

Prenatal investigation of a 45,X/46,X,r(?) karyotype in amniocytes using fluorescence in situ hybridization with an X-centromeric probe

The karyotype of cultured amniotic fluid cells obtained on the indication of advanced maternal age was shown to be a mosaic 45,X/46,X,r(?). The small size and banding pattern made it difficult to determine whether the ring was derived from and X or a Y chromosome, or even from an autosome. By using an X-centromeric probe and fluorescence in situ hybridization (FISH), we demonstrated the ring to have an X centromere. Thus, a more complete genetic counselling was possible. This confirms the usefulness of FISH in identifying and characterizing this and other chromosome rearrangements in prenatal diagnosis.     

Prenatal sex diagnosis: Testosterone and fsh levels in mid-trimester amniotic fluids

We studied the validity of mid-trimester amniotic fluid testosterone and FSH in the diagnosis of fetal sex in abnormal fetuses and in twins. It was found that the testosterone/FSH ratio, but not the level of the individual hormones, was an excellent criterion for fetal sex diagnosis. In a control group of 32 female and 34 male fetuses, the testosterone/FSH ratio was diagnostic. In two cases the values were indeterminate and fetal sex could not be predicted, but there were no false diagnoses. The abnormalities of the fetuses were diagnosed by karyotype analyses or raised levels of α-fetoprotein. It was found that neither autosomal trisomies nor X-chromosome aneuploidy or neural tube defect interfered with the diagnostic value of the hormonal approach. The twin study included 14 pairs, three of whom were discordant for sex. In all twins the fetal sex diagnosis was appropriate. In co-twins of the same sex the hormone levels were very close to each other.     

Ultrafine particles at workplaces of a primary aluminium smelter

The number concentration and size distribution of ultrafine particles in a S?derberg and a prebake potroom of an aluminium primary smelter have been measured using a scanning mobility particle spectrometer. The particle morphology was studied by transmission electron microscopy (TEM). The study shows the existence of elevated number concentrations of ultrafine particles in both potrooms. The main source of these particles is likely to be the process of anode changing. The ultrafine particles were measured directly at the source but could also be identified as episodes of high number concentrations in the general background air. Unlike the larger particles belonging to the 50-100 nm mode, the nanoparticle mode could not be detected in the TEM indicating that they may not be stable under the applied sampling conditions and/or the high vacuum in the instrument.     

Plant and Soil System Responses to Ozone After 3 Years in a Lysimeter Study with Juvenile Beech (Fagus sylvatica L.)

A lysimeter study was performed to monitor effects of elevated ozone on juvenile trees of Fagus sylvatica L. as well as on the plant–soil system. During a fumigation period over almost three growing seasons, parameters related to plant growth, phenological development and physiology as well as soil functions were studied. The data analyses identified elevated ozone to delay leaf phenology at early and to accelerate it at late developmental stages, to reduce growth, some leaf nutrients (Ca, K) as well as some soluble phenolics (hydroxycinnamic acid derivatives, total flavonol glycosides). No or very weak ozone effects were found in mobile carbon pools of leaves (starch, sucrose), and other phenolic compounds (flavans). Altered gene expression related to stress and carbon cycling corresponded well with findings from leaf phenology and chemical composition analyses indicating earlier senescence and oxidative stress in leaves under elevated ozone. Conversely in the soil system, no effects of ozone were detected on soil enzyme activities, rates of litter degradation and lysimeter water balances. Despite the fact that the three reported years 2003–2005 were climatically very contrasting including a hot and dry as well as an extremely wet summer, and also mild as well as cold winters, the influence of ozone on a number of plant parameters is remarkably consistent, further underlining the phytotoxic potential of elevated tropospheric ozone levels.