Geochemical evidence for the formation of a large miocene “travertine” mound at a sublacustrine spring in a soda lake (Wallerstein castle rock, nördlinger ries, Germany)

Summary “Travertines” (tufa pinnacles) of the Miocene Riescrater basin have been investigated to test whether carbon, oxygen and strontium isotopes can be used for the recognition of fossil subaquatic spring deposits in high-alkalinity settings. The Ries basin “travertines” have so far been interpreted as a product of subaerial to sublacustrine artesian springs discharging calcareous groundwater into a freshwater or slightly saline lake. However, recent studies on microfacies and fabric development propose a formation at Ca²⁺-supplying sublacustrine springs of a soda lake. Geochemical analysis of “travertines” of the castle rock Wallerstein, including “sickle-cell” limestones, thrombolites, non-skeletal stromatolites, and speleothems, now support the latter interpretation. High Sr contents surpassing that of the contemporaneously formed dolomitic algal biocherms of the lake shore point to an aragonitic composition of primary precipitates. the δ¹³C values of diagenetically moderately to weakly altered “travertine” facies types are in the same range of the impact-brecciated Upper Jurassic limestones, thus, are inconsistent with a mixture of soil-derived CO₂ and CO₃ ²⁻ from the Jurassic limestones. In addition, the δ¹⁸O values are too high to support a significant contribution of CO₃ ²⁻ by meteoric waters seeping through marine Jurassic limestones. Instead the δ¹³C and δ¹⁸O values indicate an origin of the CO₃ ²⁻ from a lake water body characterized by evaporation. This is consistent with a sodium-rich lake water as indicated by high sodium contents of aragonitic algal bioherms of the lake shore. The⁸⁷Sr/⁸⁶Sr isotope ratio of the “travertine” mound carbonates are consistent with calculated mixing of spring waters discharging from the crystalline basement and lake water high in dissolved inorganic carbon. This points to an origin of the divalent cations from sublacustrine spring waters. In turn,⁸⁷Sr/⁸⁶Sr isotope ratios of green algal reef carbonates of the lake shore are closer to that of the Upper Jurassic carbonates, due to surface run-off from surrounding limestone uplands.     

The Distribution and Neurochemistry of the Parathyroid Hormone 2 Receptor in the Rat Hypothalamus

This study reports the distribution of parathyroid hormone 2 receptor (PTH2R)-immunoreactive fibers in the hypothalamus using fluorescent amplification immunocytochemistry. The pattern of immunolabeling is strikingly similar to that of tuberoinfundibular peptide of 39 residues (TIP39), a peptide recently purified from bovine hypothalamus and proposed to be a ligand of the PTH2R based on pharmacological data. To investigate the anatomical basis of suggestions that TIP39 affects the secretion of several hypophysiotropic hormones we performed double-labeling studies and found that only somatostatin fibers contain PTH2R in the median eminence, which suggests that somatostatin release could be directly regulated via the PTH2R. However, several hypothalamic nuclei projecting to the median eminence contain a high density of both TIP39 and PTH2R fibers and terminals. We report here, that the PTH2R terminals also contain vesicular glutamate transporter−2, and suggest that TIP39 terminals are ideally positioned to modulate glutamatergic influences on hypophysiotropic neurons.Special Issue Dedicated to Miklós Palkovits.     

Method for determining the spatial position of the shoulder with ultrasound-based motion analyzer.

Several methods have been developed recently for the analysis of the spatial motion of the scapula and the arm, whereby the spatial position of shoulder bones is determined in static conditions by interrupting motion. The authors have developed a 3D motion analysis method recording scapular motion in progress with appropriate accuracy in the course of arm movements of various degrees. The objective of this study is to explore the applicability of the method developed, as well as to compare it with and verify it by other methods developed earlier. The position and displacements of shoulder bones were determined on 30 shoulders of 15 healthy people. The newly developed measurement method is based on the mechanical basic principle stating that the position and motion of a rigid body -- in this case, the bones (segments) forming the shoulder joint -- can be calculated at any moment from the spatial coordinates of three points of a segment and any changes thereof in the course of motion. Ultrasound-based triplets providing the three points (fundamental points) by a segment as required for measurement were fixed on the sternum (modeling the trunk), the clavicle, the acromion (modeling the scapula), the upper arm, and the lower arm. The position of the sixteen anatomical points involved in the study were determined by an ultrasound-based pointer in the local coordinate system specified by the fundamental points before starting measurements. The ZEBRIS ultrasound-based motion analysis system was used for measuring the spatial coordinates of triplets in the course of continuous motion. The spatial coordinates of the designated anatomical points can be calculated by the method of triangulation. The method was calibrated by a ZEBRIS mapping (3DCAD) software commercially available, and the measurement error rate of the method was determined by statistical calculations. On the basis of calibration and error calculations it could be established that the accuracy and the reproducibility of the method were appropriate, in accordance with the limit values to be found in the literature.     

Nitrification and degradation of halogenated hydrocarbons—a tenuous balance for ammonia-oxidizing bacteria

The process of nitrification has the potential for the in situ bioremediation of halogenated compounds provided a number of challenges can be overcome. In nitrification, the microbial process where ammonia is oxidized to nitrate, ammonia-oxidizing bacteria (AOB) are key players and are capable of carrying out the biodegradation of recalcitrant halogenated compounds. Through industrial uses, halogenated compounds often find their way into wastewater, contaminating the environment and bodies of water that supply drinking water. In the reclamation of wastewater, halogenated compounds can be degraded by AOB but can also be detrimental to the process of nitrification. This minireview considers the ability of AOB to carry out cometabolism of halogenated compounds and the consequent inhibition of nitrification. Possible cometabolism monitoring methods that were derived from current information about AOB genomes are also discussed. AOB expression microarrays have detected mRNA of genes that are expressed at higher levels during stress and are deemed “sentinel” genes. Promoters of selected “sentinel” genes have been cloned and used to drive the expression of gene-reporter constructs. The latter are being tested as early warning biosensors of cometabolism-induced damage in Nitrosomonas europaea with promising results. These and other biosensors may help to preserve the tenuous balance that exists when nitrification occurs in waste streams containing alternative AOB substrates such as halogenated hydrocarbons.     

Trichloroethylene degradation by butane-oxidizing bacteria causes a spectrum of toxic effects

The physiological consequences of trichloroethylene (TCE) transformation by three butane oxidizers were examined. Pseudomonas butanovora, Mycobacterium vaccae, and Nocardioides sp. CF8 utilize distinctly different butane monooxygenases (BMOs) to initiate degradation of the recalcitrant TCE molecule. Although the primary toxic event resulting from TCE cometabolism by these three strains was loss of BMO activity, species differences were observed. P. butanovora and Nocardioides sp. CF8 maintained only 4% residual BMO activity following exposure to 165 μM TCE for 90 min and 180 min, respectively. In contrast, M. vaccae maintained 34% residual activity even after exposure to 165 μM TCE for 300 min. Culture viability was reduced 83% in P. butanovora, but was unaffected in the other two species. Transformation of 530 nmol of TCE by P. butanovora (1.0 mg total protein) did not affect the viability of BMO-deficient P. butanovora cells, whereas transformation of 482 nmol of TCE by toluene-grown Burkholderia cepacia G4 caused 87% of BMO-deficient P. butanovora cells to lose viability. Together, these results contrast with those previously reported for other bacteria carrying out TCE cometabolism and demonstrate the range of cellular toxicities associated with TCE cometabolism.     

Synthesis and receptor-binding examinations of the normal and 13-epi-D-homoestrones and their 3-methyl ethers

An effective epimerization of the normal estrone 3-methyl and 3-benzyl ethers by using o-phenylenediamine and AcOH made the possibility for facile entry into the 13alpha-estrone series. Combination of this synthetic methodology with an isolation step carried out by means of the Girard-P reagent, the corresponding ethers of 13-epi-estrone were obtained in excellent yields. The 3-hydroxy and 3-methoxy D-homoestrone derivatives in both the normal and the 13alpha-estrone series were then synthesized and tested in vitro in a radioligand-binding assay. The estrogen receptor recognizes these compounds, but their relative binding affinities (RBAs) are lower than that of the reference compound 3,17beta-estradiol. The progesterone receptor-binding affinities of the four D-homo derivatives were also tested showing low values for 13alpha-D-homoestrone and its 3-methyl ether. Pharmacologically, these 13alpha-D-homoestrone derivatives are estrogen receptor-selective molecules.     

Phage annealing proteins promote oligonucleotide-directed mutagenesis in <Emphasis Type="Italic">Escherichia coli</Emphasis> and mouse ES cells

Background

The phage protein pairs, RecE/RecT from Rac or Redα/Redβ from λ, initiate efficient double strand break repair (DSBR) in Escherichia coli that has proven very useful for DNA engineering. These phage pairs initiate DSBR either by annealing or by another mechanism that is not defined.

Results

Here we report that these proteins also mediate single strand oligonucleotide repair (ssOR) at high efficiencies. The ssOR activity, unlike DSBR, does not require a phage exonuclease (RecE or Redα) but only requires a phage annealing protein (RecT or Redβ). Notably, the P22 phage annealing protein Erf, which does not mediate the same DSBR reactions, also delivers ssOR activity. By altering aspects of the oligonucleotides, we document length and design parameters that affect ssOR efficiency to show a simple relationship to homologies either side of the repair site. Notably, ssOR shows strand bias. Oligonucleotides that can prime lagging strand replication deliver more ssOR than their leading complements. This suggests a model in which the annealing proteins hybridize the oligonucleotides to single stranded regions near the replication fork. We also show that ssOR is a highly efficient way to engineer BACs and can be detected in a eukaryotic cell upon expression of a phage annealing protein.

Conclusion

Phage annealing proteins can initiate the recombination of single stranded oligonucleotides into endogenous targets in Escherichia coli at very high efficiencies. This expands the repertoire of useful DNA engineering strategies, shows promise for applications in eukaryotic cells, and has implications for the unanswered questions regarding DSBR mediated by RecE/RecT and Redα/Redβ.

     

Reversible mechanical unzipping of amyloid beta-fibrils

Amyloid fibrils are self-associating filamentous structures, the deposition of which is considered to be one of the most important factors in the pathogenesis of Alzheimer's disease and various other disorders. Here we used single molecule manipulation methods to explore the mechanics and structural dynamics of amyloid fibrils. In mechanically manipulated amyloid fibrils, formed from either amyloid beta (Abeta) peptides 1-40 or 25-35, beta-sheets behave as elastic structures that can be "unzipped" from the fibril with constant forces. The unzipping forces were different for Abeta1-40 and Abeta25-35. Unzipping was fully reversible across a wide range of stretch rates provided that coupling, via the beta-sheet, between bound and dissociated states was maintained. The rapid, cooperative zipping together of beta-sheets could be an important mechanism behind the self-assembly of amyloid fibrils. The repetitive force patterns contribute to a mechanical fingerprint that could be utilized in the characterization of different amyloid fibrils.