Changes in the accumulation of flavonoid and isoflavonoid conjugates associated with plant age and nodulation in alfalfa (Medicago sativa)

Carbohydrate and mineral nutrition was studied in relation to abscission in fruitlets from leafy inflorescences of the Washington navel orange ( Citrus sinensis [L.] Osbeck). Differences in the growth rate of the fruitlets permitted to predict abscission several weeks in advance. This allowed characterization of early differences in composition and behaviour of persisting and abscising fruitlets.
Inflorescences with persisting fruitlets accumulated more mineral elements than inflorescences with abscising fruitlets, and for the phloem-mobile elements the excess accumulation was allocated to the fruitlets. Starch accumulated in the inflorescence leaves during early fruitlet growth, and this accumulation was enhanced by the persisting fruitlets despite their higher growth rate and mobilizing ability. The relations between the fruitlets and the inflorescence leaves cannot be explained totally in terms of source sink relationships; a hormonal regulation of the leaves by the fruitlets is postulated.
Acid invertase activities and hexose concentration in the pericarp were higher in the abscising fruitlets. The lower early growth rate of these fruitlets is thus not caused by a limitation in carbohydrate supply. It seems more related to carbohydrate utilization, probably hormonally mediated, as demonstrated by the higher dependence on hormone supply for the growth in vitro of the endocarp explants.     

Low-nutrient induction of abnormal chlamydial development: A novel component of chlamydial pathogenesis?

Abstract The intracellular development of chlamydiae in McCoy cells incubated in Eagle's minimal essential medium lacking all 13 amino acids was examined both by fluorescence and electron microscopy and by infectivity titration. Aberrant development occurred in almost all inclusions of strains of Chlamydia trachomatis and C. psittaci with the production of abnormal forms which differed in size, shape and internal structure from normal reticulate and elementary body forms. Detailed analysis of the response of C. trachomatis L2 strain 434 to graded reductions in amino acid level showed that infectivity was reduced and morphological abnormality increased as amino acid concentrations were lowered from 33 to 0% of amino acids present in minimal essential medium. Reversion of inclusions to normal and reappearance of infectious forms occured on restoration of amino acids and further incubation. It is suggested that aberrant development may account for the presence in vivo of non-cultivable chlamydiae and that such development can arise via tryptophan deprivation mediated by local release of interferon gamma.     

Isolation of a putative fimbrial adhesin from Bordetella pertussis and the identification of its gene

We report the purification of a minor Bordetella pertussis fimbrial subunit, designated FimD, and the identification of its gene (fimD.) FimD could be purified from the bulk of major fimbrial subunits by exploiting the fact that major subunit-subunit interactions are more stable in the presence of SDS than minor-major subunit interactions. To locate the gene for FimD, internal peptides of FimD were generated, purified and sequenced. Subsequently, an oligonucleotide probe, based on the primary sequence of one peptide, was used to clone fimD. The primary structure of FimD, derived from the DNA sequence of its gene, showed homology with a number of fimbrial adhesins. Most pronounced homology was observed with MrkD, a fimbrial adhesin derived from Klebsieila pneumoniae. These observations suggest that FimD may represent a B. pertussis fimbrial adhesin. With a fimD-specific probe we detected the presence of a fimD homologue in Bordetella parapertussis and Bordetella bron-chiseptica but not in Bordetella avium. Cloning and sequencing revealed that the B. parapertussis and B. bronchiseptica fimD product differed from the B. pertussis fimD product in 20 and 1 amino acid residues, respectively. Since B. bronchiseptica is normally not a human pathogen, but causes respiratory disease in a wide range of non-human mammalian species, this may suggest that FimD recognizes a receptor that is well conserved in mammalian species. An in-frame deletion in fimD completely abolished FimD expression and also affected the expression of the major subunits Fim2 and Fim3 suggesting that, in contrast to other adhesins that are minor components of fimbriae, FimD is required for formation of the fimbrial structure.     

Changes in Aldose Reductase After Crush Injury of Normal Rat Sciatic Nerve

The response of aldose reductase (AR) to crush injury was studied in normal rat sciatic nerve. Enzyme activity and immunoreactivity of AR were determined at intervals of 1, 5, 14, 28, and 35 days after crush and correlated with histologic and immunocytochemical observations. During nerve degeneration in the distal segments of crushed nerves, a significant reduction in AR activity was detected. At 5 and 14 days, coincident with Schwann cell proliferation, enzyme activity decreased by nearly two- and fourfold, respectively. Although activity of AR increased by 28 days during nerve regeneration, it was not restored to normal levels at 35 days. Similar reductions were observed with the immunoblotting of the enzyme. Quantitative analysis of immunogold labelling on electron micrographs confirmed that proliferating as well as remyelinating Schwann cells contained reduced gold particle density compared to Schwann cells of noncrushed myelinated fibers. Immunoblots of P0, a marker for the degree of Schwann cell differentiation or myelination, showed that the temporal sequence of changes in P0 paralleled that of AR. Thus expression of AR is a function of differentiated or mature Schwann cells. The putative volume regulatory role of AR in Schwann cells may become superfluous during Wallerian degeneration.     

Glutamate and γ-Aminobutyric Acid Neurotransmitter Systems in the Acute Phase of Maple Syrup Urine Disease and Citrullinemia Encephalopathies in Newborn Calves

Cerebral cortex tissue was obtained at autopsy from neonatal Poll Hereford calves with clinically confirmed maple syrup urine disease (MSUD), neonatal Holstein-Friesian calves with clinically confirmed citrullinemia, and matched controls. From this, synaptosomes were prepared for studies of neurotransmitter amino acid uptake and stimulus-induced release, and synaptic plasma membranes were obtained for studies of associated postsynaptic receptor binding sites. As well as having abnormal brain tissue concentrations of the pathognomic plasma amino acids (markedly increased levels of the branched-chain compounds valine, isoleucine, and leucine in MSUD; marked elevation of citrulline levels in citrullinemia), both groups of diseased animals showed reduced brain tissue concentrations of each of the transmitter amino acids glutamate, aspartate, and gamma-aminobutyric acid (GABA). Nontransmitter amino acids were generally unaffected in either disease. Citrullinemic calves showed a marked increase in brain glutamine concentration; in calves with MSUD, the glutamine concentration was raised, but to a much lesser extent. The Na(+)-dependent synaptosomal uptake of both glutamate and GABA was markedly reduced (to less than 50% of control values in both cases) in citrullinemic calves but was unaltered in calves with MSUD. Whereas synaptosomes from normal calves showed the expected stimulus-coupled release of transmitter amino acids, especially glutamate and aspartate, and no response to stimulus of nontransmitter amino acids, there was no increased release of transmitter amino acids in response to depolarization in synaptosomes from citrullinemic calves.(ABSTRACT TRUNCATED AT 250 WORDS)     

Algal responses to experimental nutrient addition in the littoral community of a subtropical lake

1. An in situ experiment was performed in the littoral zone of a large, subtropical lake to quantify effects of phosphorus (P) and nitrogen (N) on algal biomass, productivity, nutrient content and phosphate uptake kinetics. 2. We hypothesized that resident periphyton rapidly sequester added nutrients from the water column, but once a certain threshold is reached, nutrients remain in the water and permit a shift to a phytoplankton-dominated community. 3. Three duplicate sets of 1.2-m diameter mesocosms were treated with 10, 20 or 50 μg P L^??1 in combination with 100, 200 or 500 μg N L^??1, respectively. The nutrients were added thrice weekly for 14 days, after which the treatment doses were doubled for an additional 9 days. The cumulative amounts of P and N added over the course of the study were 700 and 7000 μg L^??1, respectively. Two untreated mesocosms and two open reference sites were used as controls. 4. The total P concentration in the water column of nutrient-treated mesocosms remained low, even after prolonged high dosing. However, there was a two-fold increase in the P content of surface algal mats and epiphyton. This indicates that some of the added P was sequestered by those components of the community. In contrast, metaphyton and epipelon displayed little or no increase in their P content. Large quantities of added P could not be accounted for in the periphyton community, and may reflect unmeasured losses to the sediments or other pools. 5. Nitrogen also was depleted from the water column, but there were no significant increases in periphyton N content. Much of the added N could not be accounted for in mass balances, and may have been lost from the mesocosms through volatilization or other biochemical processes. 6. Chlorophyll-a in epiphyton increased significantly after 14 days in the highest nutrient treatment, where there also was a proliferation of Spirogyra on day 28. 7. On day 28, water column samples from the highest nutrient treatment also displayed a significantly higher rate of carbon uptake, and a significantly higher concentration of midday dissolved oxygen. 8. The hypothesis that phytoplankton become dominant at high nutrient loading rates was not supported. However, there were dramatic changes in community structure (increased dominance by epiphytic Spirogyra) and function (increased productivity and dissolved oxygen) in response to nutrient additions.     

The effects of reduced and elevated CO2 and O2 on the seaweed Lomentaria articulata

We grew a non-bicarbonate using red seaweed, Lomentaria articulata (Huds.) Lyngb., in media aerated with four O₂ concentrations between 10 and 200% of current ambient [O₂] and four CO₂ concentrations between 67 and 500% of current ambient [CO₂], in a factorial design, to determine the effects of gas composition on growth and physiology. The relative growth rate of L. articulata increased with increasing [CO₂] up to 200% of current ambient [CO₂] but was unaffected by [O₂]. The relative growth enhancement, on a carbon basis, was 52% with a doubling of [CO₂] but fell to 23% under 5× ambient [CO₂]. Plants collected in winter responded more extremely to [CO₂] than did plants collected in the summer, although the overall pattern was the same. Discrimination between stable carbon isotopes (Δ¹³C) increased with increasing [CO₂] as would be expected for diffusive CO₂ acquisition. Tissue C and N were inversely related to [CO₂]. Growth in terms of biomass appeared to be limited by conversion of photosynthate to new biomass rather than simply by diffusion of CO₂, suggesting that non-bicarbonate-using macroalgae, such as L. articulata, may not be directly analogous to C3 higher plants in terms of their responses to changing gas composition.     

Taxonomic and structural similarities in soil oribatid communities

We examined the taxonomic and structural concordance between oribatid mite communities at different sites from around the world using meta-analysis. The study explored similarities in the taxonomie composition and species abundance distributions among oribatid mite communities using 25 published species lists from Africa, Australia, Europe. Japan. Malaysia, New Zealand and the United States, The final data set contained 253 genera from 97 families in 43 superfamilies, A second data set, which included only studies where trees were present at the sites, contained 234 genera, 93 families and 41 superfamilies. Composition was analysed at taxonomic levels higher than species. Only two families and four superfamilies were present in > 90% of all studies. Two genera, six families and seven superfamilies were found in all sites with trees. Common families were different among habitats suggesting that habitat preferences may be expressed at the family level. There were very strong relationships between oribatid mite species richness and richness at higher taxonomic levels with r² values for regression relationships > 0.8. Few species at a site were from the same genus, family or superfamily. Species abundance distributions were remarkably consistent between studies with the three most dominant species each constituting > 10% of the oribatid individuals. Overall, similar factors may determine the relative proportions of soil-dwelling oribatid species at a site.     

False positive diagnosis of meningococcal infection by the IS1106 PCR ELISA

At a time when optimal case ascertainment for meningococcal infection is a high priority, the need for non-culture case confirmation, in particular by DNA amplification, is seen as being of vital importance to assist contact management and cluster recognition. A solution hybridisation assay with colorimetric microtitre plate detection (polymerase chain reaction-enzyme-linked immunosorbent assay (PCR ELISA)) has been developed using the multicopy insertion sequence IS1106 which had reportedly achieved a specificity of 100% and was described as being meningococcal specific. This PCR ELISA assay was evaluated on specimens from over 5000 patients at the national Meningococcal Reference Unit (MRU) between late 1995 and early 1997 and was found to be highly sensitive. Insertion sequences, however, are genetically mobile with the ability to spread between species and even genera. During the evaluation period of the IS1106 PCR ELISA a number of false positives proved to be caused by organisms other than N. meningitidis were recorded resulting in the withdrawal of this assay as a front line screening assay for routine confirmation of meningococcal infection.     

Differentiation of intestinal spirochaetes by multilocus enzyme electrophoresis analysis and 16S rRNA sequence comparisons

Abstract Multilocus enzyme electrophoresis (MEE) analysis and comparisons of nearly complete 16S rRNA gene sequences (1416 nucleotide positions) were used to evaluate phylogenetic relationships among Serpulina hyodysenteriae strain B78^T, S. innocens strain B256^T, Brachyspira aalborgi strain 513A^T, and eight uncharacterised strains of swine, avian, and human intestinal spirochaetes. From MEE analysis, nine strains could be assigned to five groups containing other intestinal spirochaetes ( genetic distances between groups = 0.6–0.9). Chicken spirochaete strain C1 and B. aalborgi 513A^T represented unique electrophoretic types and formed their own MEE groups. Despite MEE differences, the 11 strains had highly similar (96.3–99.9%) 16S rRNA sequences. These findings point out limitations of both MEE analysis and 16S rRNA sequence comparisons when used as solitary techniques for classifying intestinal spirochaetes related to Brachyspira/ Serpulina species.