Similarities among nitrogenase proteins and amongnif-HDK genes ofMethanosarcina barkeri and of other diazotrophic bacteria

Similarities between the nitrogen-fixing systems of the archaebacteriumMethanosarcina barkeri (strain Fusaro) and a number of eubacteria were investigated. Using antibodies againstRhizobium leguminosarum nitrogenase and a probe of clonednif-HDK genes of this species, homology withM. barkeri was demonstrated on the protein level and to a greater extent on the DNA level.     

Spatial heterogeneity in the relative impacts of foliar quality and predation pressure on red oak, Quercus rubra, arthropod communities

Predation pressure and resource availability often interact in structuring herbivore communities, with their relative influence varying in space and time. The operation of multiple ecological pressures and guild-specific herbivore responses may combine to override simple predictions of how the roles of plant quality and predation pressure vary in space. For 2 years at the Coweeta LTER in the Southern Appalachian Mountains, we conducted a bird exclosure experiment on red oak (Quercus rubra) saplings to investigate the effects of bird predation on red oak arthropod communities. We established bird exclosures at six sites along an elevational gradient and estimated variation in foliar nitrogen and bird predation pressure along this gradient. Foliar nitrogen concentrations increased with elevation while our index of bird predation pressure was variable across sites. Greater arthropod densities were detected inside exclosures; however, this result was mainly driven by the response of phloem feeders which were much more prevalent inside exclosures than on control trees. There was little evidence for an effect of bird predation on the other arthropod guilds. Consequently, there was no evidence of a trophic cascade either in terms of leaf damage or tree growth. Finally, we found more variation in arthropod density among trees within sites than variation in arthropod density among sites, indicating the importance of micro-site variation in structuring arthropod communities.     

Bacteroides xylanolyticus sp. nov., a xylanolytic bacterium from methane producing cattle manure

As part of a study of the biogas production from cattle waste, xylanolytic bacteria were isolated from enrichments of fermenting cattle manure. From 34 isolates, mostly Gram-negative rods, a typical strain was investigated in more detail. It was an anaerobic non-sporeforming, Gramnegative rod, which was motile with peritrichous flagella. This organism fermented xylan and many soluble sugars (glucose, cellobiose, mannose, xylose, arabinose). Other hemicelluloses such as gum xanthan, laminaran, locust bean gum, and gum arabic were not utilized. It also could not use cellulose. Fermentation products were carbon dioxide, hydrogen, acetate and ethanol. The bacterium produced carboxymethylcellulase and xylanase, especially when growing on xylan. Growth was optimal between 25°C and 40°C and between pH 6.5 and 7.5. The guanine plus cytosine content of the DNA was 34.8±0.8%. The isolate was identified as a member of the genus Bacteroides, and a new species is proposed: Bacteroides xylanolyticus (xylan dissolving). The type strain of B. xylanolyticus is strain X5-1 (DSM 3808).     

Biodegradation of 4-nitroanisole by two Rhodococcus spp.

Two Rhodococcus strains, R. opacus strain AS2 and R. erythropolis strain AS3, that were able to use 4-nitroanisole as the sole source of carbon and energy, were isolated from environmental samples. The first step of the degradation involved the O-demethylation of 4-nitroanisole to 4-nitrophenol which accumulated transiently in the medium during growth. Oxygen uptake experiments indicated the transformation of 4-nitrophenol to 4-nitrocatechol and 1,2,4-trihydroxybenzene prior to ring cleavage and then subsequent mineralization. The nitro group was removed as nitrite, which accumulated in the medium in stoichiometric amounts. In R. opacus strain AS2 small amounts of hydroquinone were produced by a side reaction, but were not further degraded.     

Enzyme profiles of Thiobacillus versutus after aerobic and denitrifying growth: Regulation of isocitrate lyase

The specific activities of the tricarboxylic acid (TCA) cycle enzymes in Thiobacillus versutus were invariably lower after aerobic growth as compared to denitrifying growth in acetate- or succinate-limited chemostat cultures. Of the glyoxylate cycle enzymes, isocitrate lyase (ICL) activity was nil during aerobic and 76 nmol·min^-1·mg^-1 protein during denitrifying growth on acetate whereas malate synthase (MS) did not change. In succinate-grown cells ICL was always near nil. The change in ICL and MS was followed after pulse additions of acetate and nitrate to an aerobic acetate-limited chemostat culture made anaerobic prior to the first pulse. ICL remained nil during denitrifying growth after the first pulse but increased to 47 and 81 nmol ·min^-1·mg^-1 protein after the second and third pulse, respectively. MS remained unaltered. The appearance of ICL was dependent upon de novo protein synthesis. During transition in a steady state culture on acetate from oxygen to nitrate as terminal electron acceptor, denitrifying growth started after 0.6 volume replacements. The resumption of growth was concomitant with an increase in TCA cycle enzyme activities. ICL was observed only after two volume replacements. During the reverse transition, ICL disappeared at a rate twice the dilution rate. SDS polyacrylamide gelectrophoresis of cell-free extracts containing ICL showed a major protein band with a R_f value identical to purified ICL and a mol·wt. of 60,000. ICL from T. versutus was inhibited by 1.5 mM itaconate but not by 10 mM phosphoenolpyruvate. Its activity was dependent upon the presence of Mg²⁺ and cysteine.Abbreviations TCA Tricarboxylic acid - ICL isocitrate lyase - MS malate synthase - FPLC fast protein liquid chromatography - maximum specific oxygen consumption rate     

Arterial Expression of the Calcium-Sensing Receptor Is Maintained by Physiological Pulsation and Protects against Calcification

Vascular calcification (VC) is common in chronic kidney disease (CKD) and contributes to cardiovascular mortality. The calcium-sensing receptor (CaSR) is present in human artery, senses extracellular calcium and may directly modulate VC. Objective: to investigate the association between arterial cyclic strain, CaSR expression and VC. Methods and Results: human aortic smooth muscle cells (HAoSMC) were cultured under static or strained conditions, with exposure to CaSR agonists, the calcimimetic R568, and after CaSR silencing and over-expression. High extracellular calcium reduced CaSR expression and promoted osteochondrogenic transformation and calcium deposition. This was partially prevented by cyclic strain and exposure to R568. CaSR silencing enhanced calcification and osteochondrogenic transformation, whereas CaSR over-expression attenuated this procalcific response, demonstrating a central role for the CaSR in the response to cyclic strain and regulation of VC. In arterial explants from CKD patients (n = 11) and controls (n = 9), exposure to R568 did not significantly alter calcium deposition, osteochondrogenic markers or total artery calcium content. Conclusions: physiological mechanical strain is important for arterial homeostasis and may protect arteries from VC. The beneficial effects of cyclic strain may be mediated via the CaSR.     

Isolated Follicles Enriched for Centroblasts and Lacking t(14;18)/BCL2 in Lymphoid Tissue: Diagnostic and Clinical Implications

We sought to address the significance of isolated follicles that exhibit atypical morphologic features that may be mistaken for lymphoma in a background of reactive lymphoid tissue. Seven cases that demonstrated centroblast-predominant isolated follicles and absent BCL2 staining in otherwise-normal lymph nodes were studied. Four of seven cases showed clonal B-cell proliferations amid a polyclonal B cell background; all cases lacked the IGH-BCL2 translocation and BCL2 protein expression. Although three patients had invasive breast carcinoma at other sites, none were associated with systemic lymphoma up to 44 months after diagnosis. The immunoarchitectural features of these highly unusual cases raise the question of whether a predominance of centroblasts and/or absence of BCL2 expression could represent a precursor lesion or atypical reactive phenomenon. Differentiating such cases from follicular lymphoma or another mimic is critical, lest patients with indolent proliferations be exposed to unnecessarily aggressive treatment.     

Behaviour of toluene, benzene and naphthalene under anaerobic conditions in sediment columns

The biotransformation of toluene, benzene and naphthalene was examined in anaerobic sediment columns. Five columns filled with a mixture of sediments were operated in the presence of bicarbonate, sulfate, iron, manganese, or nitrate as electron acceptor. The columns were continuously percolated with a mixture of the three organic compounds (individual concentrations 25–200 μM) at 20°C. Toluene was transformed readily (within 1 to 2 months) under all redox conditions tested. Benzene was recalcitrant over the test period of 375–525 days in all five columns. Naphthalene was partly transformed in the column with nitrate or manganese as electron acceptor present; the addition of benzoate had a positive effect in the column with nitrate. In the column with sulfate, the majority of the added naphthalene disappeared. No effect was observed after adding and omitting an easier degradable substrate. [¹⁴C]naphthalene was used to confirm this disappearance to be the result of degradation; two third of the naphthalene was converted to CO₂.     

Transfer of Cesium from the Xylem to the Phloem in the Stem of Wheat

Steam-girdling experiments with detached wheat shoots showed that cesium was eliminated from the xylem sap and loaded into the phloem during acropetal transport. This transfer is important for the accumulation of cesium (especially also of the radiopollutants ¹³⁴Cs and ¹³⁷Cs) in maturing wheat grains.